CS247168B2 - Method of new carbapenem derivatives production - Google Patents

Abstract

Disclosed are novel carbapenem derivatives characterized by a 2-substituent of the formula <IMAGE> in which A represents a C1-C6 straight or branched chain alkylene group; R<5> represents an optionally substituted aliphatic, cycloaliphatic, cycloaliphatic-aliphatic, aryl, araliphatic, heteroaryl, heteroaraliphatic, heterocyclyl or heterocyclyl-aliphatic radical and <IMAGE> represents a nitrogen-containing aromatic heterocycle attached to the alkylene group A at a ring carbon atom and quaternized by substituent R<5>. Such derivatives are useful as potent antibacterial agents.

Description

(54) Method for producing new carbapenem derivatives

A process for the preparation of carbapenem derivatives of formula I and their pharmaceutically acceptable salts, which comprises reacting an intermediate of formula III to form an intermediate of formula IV, which is reacted with a mercaptan of formula I to form an intermediate of formula II, and reacted with an alkylating agent of formula to give a compound of formula 1, and optionally replacing the anion X 'for different ^ Nion and optionally removing protecting group R ² carboxyl and optionally the compound of formula I into a pharmaceutically acceptable salt thereof.

The compounds of formula I are useful for the treatment of bacterial infections.

The present invention relates to novel carbapenem antibiotics having a substituent of the general formula at position 2

--S-A where

A is C1-C6 alkylene with straight or branched chain,

R ⁵ is an optionally substituted aliphatic, cycloaliphatic, cycloaliphatic-aliphatic, aryl, arylaliphatic, heteroaryl, heteroaryl aliphatic, heterocyclic or heterocyclic-aliphatic residue and the ring of formula represents a nitrogen-containing aromatic heterocycle attached to alkylene group A by a ring carbon atom and quaternized.

A number of beta-lactam derivatives containing a carbapenem nucleus are described in the literature

These carbapenem derivatives are beta-lactamase binders.

useful as antibactericidal agents and / or inhibitors of natural products such as thienamycin of the formula

Original carbapenem compounds

OH

COOH .sch ₂ ch ₂ nh ₂ and were prepared by fermentation of Streptomyces cattleya (U.S. Pat. No. 3,950,357). Thienamycin is an exceptionally potent broad-spectrum antibiotic that is particularly effective on various species of Pseudomonas, organisms known to be resistant to beta-lactam antibiotics.

Other natural carbapenem-containing compounds include olivanoic acid derivatives such as the antibiotic MM 13902 of the formula

HO3SO

claimed in US patent 4 113 856, the antibiotic MM 17880 formulas HO3SO ' CH ₃ Vn _x SCH ₂ CH ₂ NHCOCH ₃ ^ COOH claimed in US patent 4 162 304, antibiotic MM 4550A formula CH ₃ O 1 гS-CH = CHNHCOCH ₃ HO ₃ SO ^Z 0 COOH claimed in US patent C, . 4 172,129, a antibiotic 890A ~ formula CH3 0 J) _z x = CHNHCCH-SCH ₃ HO ₃ SO ^X jaO 0 COOH claimed in US patent 4 264 735.

In addition to natural compounds, the compound desacctyl 890A formula

It is disclosed in U.S. Patent 4,264,734 and is prepared by enzymatic deacylation of the corresponding N-acetyl compound. Various derivatives of natural olivanoic acids, for example compounds of the formula, have also been synthesized

^S °> n \ = _/ ^NHCOCH 3 wherein CO2Rj is free, esterified n is 0 or 1 and _R? is hydrogen, carboxyl acyl

A COjR-group or a carboxyl group in the form of a salt, or a group of the formula

wherein R2 is a salt-forming ion or methyl or ethyl.

These compounds are disclosed in European Published Patent Application 8885.

U.S. Pat. No. 4,235,922 (see also European Carbapenem formula published patent application 2058) claims

SCH ₂ CH ₂ NH ₂

COOH whereas British published patent application 1,598,062 claims the isolation of sch ₂ ch ₂ ncoch ₃ ox COOH from a Streptomyces fermentation medium.

Carbapenems, which are unsubstituted at the 6-position, were also synthesized. Thus, U.S. Pat. No. 4,210,661 discloses compounds of the formula.

S-R2

COOH wherein R 2 is phenyl or substituted phenyl, US Patent 4,267,177 discloses compounds of formula

S-R-1

COOH wherein R 8 is an optionally substituted pyridyl group and US Patent 4,255,441 discloses compounds

S - CR 2 - CR 3 R 4

COOH wherein R 2 and R 1 are hydrogen or alkyl and R 8 is bolded phenyl and n is 1 or 2 and US Patent 4,282

NH-CO 2 R 8, wherein R 8 is alkyl, phenyl or substi236 discloses compounds of formula

Wherein R ^ is hydrogen and R ^ is cyano or CC ^ R ^ where hydrogen, alkyl, aryl or arylalkyl.

Carbapenems of general formula

wherein R is hydrogen or acyl and R is hydrogen or substituted or unsubstituted alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, alkylcycloalkyl, aryl, arylalkyl, arylalkenyl, arylalkinyl, heteroaryl heteroarylalkyl, heterocyclyl or heterocyclylalkyl are disclosed in U.S. Pat. 218 463. There are no claims for any heteroarylalkyl substituents of the R ⁸ type

wherein A is alkylene and part of the molecule

is a quaternary nitrogen containing aromatic heterocycle attached to alkylene group A via a carbon atom in the ring.

The natural product thienamycin has an absolute configuration of 5R, 6S, 8R. This isomer as well as the remaining seven thienamycin isomers can be obtained by the total synthesis disclosed in U.S. Patent No. 4,234,596. Total synthesis procedures are also disclosed, for example, in the US patents

287 123, 4,269,772, 4,282,148, 4,273, 7973. A key intermediate

709, 4,290,947 and in published European patent in the claimed synthetic methods is where p-nitrobenzy1 is.

CO ₂ pNB

Due to the exceptional biological activity of thienamycin, a large number of derivatives have been prepared and documented in the literature. Of these compounds are

1) N-formimidoylthienamycin of formula

OH sch ₂ ch ₂ n = c-nh ₂

H

Q COOH is exemplified in European Published Patent Application 6639

2) N-he terocyclic derivatives of t. Hienamycin of formulas I and II

where

) Bifunctional ring may contain additional unsaturation in the ring AA ^from 6 ^pj 0, ¹ or 2, ^R1 is hydrogen, amino or alkyl, is hydrogen, alkyl or aryl and are exemplified in U.S. Patent 4,189,493 wherein n is an integer selected Z is imino, oxo

3) substituted thienamycin N-methylenderivates of the general formula

sch ₂ ch ₂ n = cx

Y

1 _of 1 2 X and Ύ are hydrogen, R, OR, SR or NR R

COOH)

wherein wherein alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, arylalkyl, heterocyclyl or heterocyclylalkyl and R ¹ and ny in U.S. Patent 4,194,047,

R is substituted or unsubstituted aryl, aryLalkyl, heteroaryl, heteroR are hydrogen or R is lower4) compounds of formula

SCH ₂ CH ₂ NR ¹ R ²

COOH where R ³ is aryl, alkyl, acyl or arylalkyl from hydrogen and acyl (including type 0 acyl)

-c R and R are each independently selected ^{R 11} wherein ^{R! Have 1} m e ^{i b} j yt limit ^by unique quaternary ammonium groups, for example substituted alkyl are disclosed in U.S. Patent

5) compounds of formula

sch ₂ ch ₂ nr ¹ r ²

_From COOH wherein R is _a hydrogen atom, or a monovalent acyl, R1 is optionally substituted alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, alkyl cykloalkenyl2, cykloalkyLaiky1, aryl, arylalkyl, heteroaryl or heteroarylalkyl, and R is acyl (including acyl type

O lí

-C-R optionally substituted hydrocarbon residue, wherein R is alkyl substituted with a quaternary ammonium group, e.g.

O n 6

-C-CH 2 are disclosed in British Patent 1,604,276 (see also US Patent 4,235,917)

6) compounds of the general formula

OH

nr ⁵ r ⁶ r ⁷ nr ⁵ r ⁶ r ⁷ where ^{r5, r6} and ^R ? are independently selected from the group consisting of hydrogen, substituted or unsubstituted alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkenylalkyl, cycloalkylalkyl, aryl, arylalkyl, heteroaryl or heteroarylalkyl are disclosed in U.S. Patent 4,235,920;

7) compounds of formula

OR ³

O cox

Wherein R and R independently of the others are radicals of the type defined for R, hydrogen or nitro, hydroxyl, alkoxy of 1 to 6 carbon atoms, amino, alkylamino of 1 to 6 carbon atoms, dialkylamino or trialkylamino of 1 to 6 carbon atoms in 12 in each of the alkyls and the other anion present in the latter case or R and R may be taken together to form, together with the nitrogen atom to which they are attached, a substituted or unsubstituted monocyclic or bicyclic heteroaryl or heterocyclic radical containing 4 to 10 ring atoms, one or more of which may be further heteroatoms selected from oxygen, sulfur and nitrogen, R is cyano or substituted or unsubstituted carbamoyl, carboxyl, alkoxycarbonyl of 1 to 11 carbon atoms, alkyl of 1 to 10 carbon atoms, alkenyl of 2 to 10 carbon atoms, alkynyl of 2 to 10 carbon atoms, cycloalkyl of 3 to 10 carbon atoms, cycloalkylalkyl of 4. C 12 -C 12 cycloalkenyl, C 5 -C 12 cycloalkenylalkenyl, C 4 -C 12 cycloalkenylalkyl, C 6 -C 10 aryl, C 6 -C 16 arylalkyl, C 8 -C 8 arylalkenyl C 1 -C 16 arylalkinyl or C 1 -C 16 arylalkynyl or monocyclic or bicyclic heteroaryl, heteroarylalkyl, heterocyclyl or heterocyclylalkyl of 4 to 10 ring atoms, one or more of which are heteroatoms selected from oxygen, sulfur and nitrogen, and wherein the alkyl radical heteroarylalkyl or heterocyclylalkyl radical contain from 1 to 6 carbon atoms, substituent

2 or the substituents on R, R and R are chlorine, bromine, iodine, fluorine, azido, C 1 -C 4 alkyl, mercapto, sulfo, phosphono, cyanthio (-SCN), nitro, cyano, amino, hydrazino, substituted amino- or hydrazino groups having at most three alkyl groups having 1 to 6 carbon atoms, hydroxyl, alkoxy groups having 1 to 6 carbon atoms, alkylthio groups having 1 to 6 carbon atoms, carboxyl, oxo, alkoxycarbonyl having 1 to 6 carbon atoms in alkoxy, acyloxy groups with 2 up to 10 carbon atoms, carbamoyl, alkylcarbamoyl of 1 to 4 carbon atoms in alkyl or dialkylcarbamoyl of 1 to 4 carbon atoms in alkyl, R7 is hydrogen, acyl or a radical of the type defined for R, R is alkyl of 1 to 10 carbon atoms, substituted carbonylmethyl, alkoxyalkyl of 1 to 6 carbon atoms in both substituent groups, cycloalkoxyalkyl of 3 to 6 carbon atoms in cycloalkoxy and of 1 to 6 carbon atoms in alkyl, alkanoyloxyalkyl of 2 to 12 carbon atoms, parts or halogenated C1-C6 alkyl wherein the halogen atoms are chlorine, bromine or fluorine, aminoalkyl, alkenyl or alkynyl of 2 to 10 carbon atoms, acyl, alkoxycarbonylalkyl of 3 to 14 carbon atoms, dialkylaminoacetoxyalkyl of 4 to 21 atoms carbon, alkanoylaminoalkyl of 2 to 13 carbon atoms, arylalkyl of 1 to 3 carbon atoms in alkyl and wherein the aryl radical contains from 6 to 10 carbon atoms, monocyclic or bicyclic heteroarylalkyl or heterocyclylalkyl of 4 to 10 ring atoms, 1 to 3 carbon atoms in the alkyl radical and 1 to 4 heteroatoms selected from oxygen, sulfur and / or nitrogen atoms, a nucleosubstituted arylalkyl or heteroarylalkyl wherein the substituent is a chlorine, fluorine, bromine or iodine atom or an alkyl of 1 to 6 carbon atoms, aryl or a nucleus substituted aryl of 6 to 10 carbon atoms and wherein any substituent on the nucleus is hydroxyl, alkyl of 1 to 6 carbon atoms, chlorine, fluorine or bromine, arylalkoxyal alkyl, thioalkyl of 2 to 12 carbon atoms, cycloalkylthioalkyl of 4 to 12 carbon atoms, acylthioalkyl of 2 to 10 carbon atoms in the acyl moiety and 1 to 6 carbon atoms of the alkyl or phenylalkenyl wherein the alkenyl has 2 to 6 atoms R 5 is substituted or i-substituted alkyl having 1 to 10 carbon atoms, alkenyl having 2 to 10 carbon atoms, or alkynyl having 2 to 10 carbon atoms; up to 10 carbon atoms on the ring

Ί substituted or unsubstituted cycloalkyl, cycloalkenyl, cycloalkenylalkyl and cycloalkylalkyl of 3 to 6 carbon atoms and up to 6 carbon atoms in any chain, aryl of 6 to 10 carbon atoms, arylalkyl of 6 to 10 carbon atoms and 1 to 6 carbon atoms in monocyclic or a bicyclic heteroaryl or heteroarylalkyl containing 4 in the ring, one or more of which is an oxygen, nitrogen or sulfur atom and 1 to the alkyl moiety and the ring or chain substituents are chlorine, ru, azido, cyano, amino, alkylamino of 1; up to 6 carbon atoms, dialkylamino with alkyl moieties, up to 10 carbon atoms of bromine, iodine, fluoro to 6 carbon atoms in each of the alkyl or C1-C6 trialkylamine in each of the alkyl, and in the latter case an additional anion is present, hydroxyl, (C 1 -C 6) alkoxy, (C 1 -C 6) alkylthioalkyl, carboxyl, oxo, (C 1 -C 6) alkoxycarbonyl, acyloxy Kupina having 2 to 10 carbon atoms, carbamoyl, alkylcarbamoyl having 1 to 4 carbon atoms, dialkylkarbamoy1 having 1 to 4 carbon atoms in each alkyl, kyanothio (SCN) n or ^even up ^to TROS ^{on R 6} is a hydrogen-water ^ ^ and ^ xy ^ ap ^t mer ^{to the} axis up to ^i, R, O ^R, S ^R, or -NR ² R, wherein R, R ¹ and R ² are as defined above, • 4 4 4 4x is hydroxyl, mercapto, amino, acyloxy, -OR, -SR, -NHR, -NR,

L

R

OM, OQ or if the compound is in the form of a double ion, -O, in tomna A,

And, if the compound is not in the form of a double ion, it is an ion

M is a pharmaceutically acceptable cation and

Q is a protecting group as defined above is claimed

No. 604,275, and in this case is not the British patent

8) compounds of the general formula

where

attached to the nitrogen atom of the amino group of the thienamycin represents a mono- or polycyclic nitrogen atom containing a heterocyclic group and is a hydrogen atom, a substituted alkenyl, heterocyclylalkenyl, alkyl, NR ₂ , COOR, CONR ₂ , -OR patent application 21082.

or unsubstituted alkyl, aryl, arylalkenyl, heterocyclylalkyl, aryl or CN are claimed in the European

Among the compounds disclosed in US Patent 4

235

920 are compounds of formula

wherein A is a pharmaceutically acceptable anion.

The aforementioned quaternary amine derivatives are also described in Recent Advances in Beta-Lactam Antibiotics, Royal Society of Chemistry, London, 1981, pp. 240-254, wherein the antibacterial activity is on average about 1/2 to 2/3 of thienamycin. .

In addition to the above, carbapenem derivatives also have a wide range of substituents at the 6-position.

For example

1) European published patent application

408 claims compounds of formula

S «51

COOH where R 6 is a hydrogen atom, methyl or hydroxyl and a monovalent organic group including, but not limited to, heterocyclylalkyl,

2) The European published patent application claims compounds of the formula

8514

COOH wherein R 6 is an optionally substituted pyrimidinyl group and R ₂ is a hydrogen atom or a group CR 6 R 6 R 6, wherein R 6 is a hydrogen atom or a hydroxyl, R 6 is a hydrogen atom or an alkyl and R 6 is a hydrogen atom, alkyl, benzyl or phenyl or R1 and R2 together form a carbocyclic ring,

3)

European Published Application 38869 discloses compounds of formula

Wherein R, R and R are independently selected from the group consisting of hydrogen, substituted or unsubstituted alkyl, alkenyl or alkynyl of 1 to 10 carbon atoms, cycloalkyl, cycloalkylalkyl, and alkylcycloalkyl of 3 to 6 carbon atoms in the cycloalkyl ring; up to 6 carbon atoms in the alkyl moiety, aryl such as phenyl, arylalkyl, arylalkenyl and arylalkinyl wherein the aryl moiety is phenyl and the aliphatic moiety has 1 to 6 carbon atoms, heteroaryl, heteroarylalkyl, heterocyclyl and heterocyclylalkyl, wherein the substituent or substituents related to the above the residues are selected from the group consisting of

-X ° halogen (chlorine, bromine, fluorine), - OH hydroxyl -OR 4 alkoxy, aryloxyl ii

-OCNR R is carbamoyloxy

O il 12

-CNR R carbamoyl

2

-NR R is amino, amidino

-NO ₂ nitro,

Θ! !

-N (R K is a tri-substituted amino group (R is selected independently) R ^{11 1} 2

-C = NOR oximino

-SR 1-6 alkyl- and arylthio

2

-SO ₂ NR R sulfonamido

1 2 -NHCNrJr ureido

O i II 2

-R CNR amido

-CO2H carboxy group ^-CO 2 ^{R 1 to} R ^b ox ^y acrylate ^{--CR @ 1} and c ^y l

1 ₁ 1

-OCR acyloxy

-SH mercapto group 1

-SR alkyl- and arylsulfinyl

O

-SR ^1-yl-alk and Ar ^y LSU ^lf he ^yl

II o

-ON cyano

- ^N 3 azido group on the list of substituents at r ^{3, R 3} ? and ^R & are su ^b s ^tit of eluent ^{R 1} and R ^{2 kd} e mod ^h le ^d em к above are independently selected from the group consisting of hydrogen, alkyl, alkenyl, and alkynyl having 1-10 carbon atoms, cycloalkyl, cycloalkylalkyl alkylcycloalkyl having 3 to 6 carbon atoms in the cycloalkyl ring and 1 to 6 carbon atoms in the alkyl portion of the molecule, aryl such as phenyl, arylalkyl, arylalkenyl and arylalkinyl, wherein the aryl moiety is phenyl and the aliphatic moiety has 1 to 6 carbon atoms, heteroaryl, heteroarylalkyl , heterocyclyl and heterocyclylalkyl and wherein the heteroatom or heteroatoms in the above-mentioned heterocyclic moieties are selected from the group consisting of 1 to 4 oxygen, nitrogen or sulfur atoms and wherein the alkyl moieties associated with these heterocyclic moieties have 1 to 6 carbon atoms (see also published European Patent Applications 1627, 1628, 10317, 17992, 37080, 37081 and 37082).

4) European Published Patent Application 34832 discloses as antibacterial agents compounds of the formula

SR ,,

CO ₂ H ^kd e ^R1is water ^Cart with EQS »ar ^PI selected from substituents per ^hour rnujíc ^I O ^ ^OS ° ^¹H no ^b her with ^U L where s 1 s 1 wherein the alkanoyl having 1 to 6 carbon atoms , arylalkanes ^yl, aryloxyalkanoyl or arylcarbonyl, ~

wherein each of the R groups is optionally substituted.

or alk ^yl ester of 1 to 4 carbon atoms or ^ SR ³ OOOR ^3, 0C0 or ³ R ₉ ³ OOONHR

R is alkyl of 1 to 6 carbon atoms or optionally substituted benzyl and R is alkyl of up to 6 carbon atoms or optionally substituted benzyl or phenyl and R is alkyl of up to 6 carbon atoms, alkenyl of 2 to 6 carbon atoms, alkynyl of 3 to 6 atoms carbon, the triple bond is not present on the carbon atom attached to the sulfur atom, arylalkylEuropean patent application 44 170 illustrates carbapenem derivatives of the formula

wherein R ³ is hydrogen or the organic group n is 0 or 1, X is saturated or unsaturated bromine or chlorine, and R is alkyl of 1 to 6 arylalkyl of 1 to 10 carbon atoms or aryl, each of which R is substituted.

bonded via a carbon atom to a carbapenem ring, a hydrocarbon radical optionally substituted by carbon atoms, alkenyl of 2 to 6 carbon atoms: ⁴ is optionally

However, none of the compounds wherein the tetrazole ring is attached to X via a quaternized nitrogen atom, i.e. a positively charged nitrogen atom that is not attached to a hydrogen atom, is claimed in the application.

European Patent Application 38 869 cited above discloses the synthesis of carbapenem derivatives via intermediates of the general formula r r -

what r ₂ ^to de ^R6 and ^R7 have ^the meanings indicated ^above, and ·

R ² is a readily sides ^{d i} n te ^{l and} carbonisation ^b ox ^yl b ^s c ^h r ^{and s} with nothing ^to oup.

Likewise, compounds of the formula are also shown as intermediates

wherein X is described as a leaving group.

At the Gordon Research Conference on Medicinal Chemistry requested in New London, New Hampshire on August 2 - 6, materials were distributed where many carbapenem antibiotics were reported. Among the compounds listed at

page 9 of the materials ⁸ -C ( ^CH 3) 3 is a carbapen of formula which differs from the compounds of the present ring in substituent 2 is bound directly to the invention in that the sulfur atom instead of the quaternized carbon heteroaroma atom of the alkylene group;

European Published Patent Application 50 334 claims carbapenem derivatives of the general formula

wherein R ⁶ and R ⁷ are independently selected from the group consisting of hydrogen, alkyl, alkenyl, aryl and arylalkyl, A is a direct single bond linking sulfur and carbon atoms, or A is a cyclic or acyclic linking group selected from among other substituents including

The 2 alkyl, cycloalkyl, aryl, heteroaryl or heteroalkyl, R and R which define the carbamimidoyl functionality are, inter alia, independently selected from the group consisting of hydrogen, alkyl and aryl, moreover, this carbamimidoyl is characterized by cyclic structures obtained by linking two nitrogen atoms through substituents; by attachment to the linking group A, carbamimidinium residues obtained by quaternizing one of the carbamimidoyl nitrogen atoms are disclosed.

On page 12 of this application, substituents at the 2-position of the group are those of the formula NR ¹

-SAC ₍

1

NR ·.

kd ^e R ¹ js ° ^for ato ^m H, substituted and unsubstituted: alkyl, cycloalkyl, cycloalkylalkyl, alkylcycloalkyl, aryl, arylalkyl, heterocyclyl or heterocyclylalkyl and the two nitrogen atoms to participate in cyclic structures which are shown as dotted lines.

There are no specific claims for any cyclized carbamimidoyl groups containing a quaternized nitrogen atom, but page 22 lists a cyclized carbamimidoyl group of the formula

Based on the above definitions of R ^1, we do not believe that the published European patent application 50 334 generally claims any of these compounds. Since the wording in that application is so broad in relation to cyclic structures, this reference is made in the present application.

While, as mentioned above, the prior art describes carbapenem derivatives having a substituent at the 2-position of the general formula

-S-A-Het wherein A is an alkylene group Het represents a heterocyclic or heteroaromatic group, there are no claims in which carbapenems are mentioned, wherein Het is a radical of formula

wherein R ⁵ is an optionally substituted aliphatic, cycloaliphatic, cycloaliphatic-aliphatic, aryl, arylaliphatic, heteroaryl, heteroarylaliphatic, heterocyclyl or heterocyclylaliphatic residue and

represents a quaternized nitrogen containing an aromatic heterocycle attached to an alkylene atom through a ring carbon atom. As mentioned above, carbapenem having a δ- ^CH 3 -S-CH ₂ CH ₂ N - CH ₃ \ ch ₃ substituent as the 2-position substituent has been mentioned, as has carbapenem with a quaternized heteroaromatic ring bonded directly to the 2-subistuent open.

Despite the vast number of carbapenem derivatives described in the literature, there is a continuing need for new carbapenems, as known derivatives can be improved in terms of their activity spectrum, degree of activity, stability and / or toxic side effects.

The present invention relates to a new series of carbapenem derivatives having a substituent at the 2-position of the general formula

wherein A is a straight or branched alkylene having 1 to 6 carbon atoms, R ⁵ is an optionally substituted aliphatic, cycloaliphatic, cycloaliphatic-aliphatic, aryl, arylaliphatic, heteroaryl, heteroarylaliphatic, heterocyclyl or heterocyclylaliphatic residue and a residue

represents a quaternized nitrogen-containing aromatic heterocycle attached to alkylene group A via a ring carbon atom.

The present invention relates to a process for the preparation of carbapenem derivatives of the general formula I

(I) where

Q

R is a hydrogen atom; is selected from the group consisting of hydrogen, substituted or unsubstituted alkyl, alkenyl and alkynyl of up to 10 carbon atoms, cycloalkyl, cycloalkylalkyl of 3 to 6 carbon atoms in the cycloalkyl ring and 1 to 6 carbon atoms in the alkyl moiety, phenyl, arylalkyl, arylalkenyl and arylalkinyl wherein the aryl moiety is phenyl and the aliphatic moiety has 1 to 6 carbon atoms, heteroaryl, heteroarylalkyl, heterocyclyl and heterocyclylalkyl wherein the heteroaryl is a mono-, di- or polycyclic aromatic heterocyclic radical having 5 or 6 members in individual rings and the heterocyclyl is mono a di- or polycyclic saturated or unsaturated aromatic heterocyclic radical having 5 or 6 members in individual rings and wherein the heteroatom or heteroatoms in the above-mentioned moieties are selected from the group consisting of 1 to 4 oxygen, nitrogen or sulfur atoms and alkyl moieties attached to said heterocyclic the moieties have from 1 to 6 carbon atoms, wherein the substituent or substituent is C 1 -C 6; the radicals of the above-mentioned radicals are independently selected from the group consisting of alkyl of 1 to 6 carbon atoms, optionally substituted with amino, halogen, hydroxyl or carboxyl, halogen

-STEED

O

4

-OCNR R

O

4 -CNR R

4

-SO ₂ NR R

O

-NHCNR ³ R ⁴

O

И 4 R CNR

-c ° ₂ r ³

-OP (O) (OR ³⁾ (OR ⁴⁾ = 0

О 3 · -OCR

-CN

-OSO ₃ R ³

O '·' 9

-OS-R

II

O

O

3 ^/; 9

-NR S-R

O • NR C = NR

R

-NR CO ₂ R

- ^NO 2 wherein in the above substituents R is independently selected from the group consisting of hydrogen, alkyl, alkenyl and alkynyl of up to 10 carbon atoms, cycloalkyl, cycloalkylalkyl and alkylcycloalkyl of 3-6 atoms in the cycloalkyl ring and 1-6 carbon atoms in the alkyl moieties, phenyl, arylalkyl, arylalkenyl and arylalkinyl, wherein the aryl moiety is phenyl and the aliphatic moiety has 1 to 6 carbon atoms and heteroaryl, heteroarylalkyl, heterocyclyl and heterocyclylalkyl, wherein heteroaryl is a mono-, di- or polycyclic aromatic heterocyclic radical having 5 or 6 members in individual rings and heterocyclyl is a mono-, di- or polycyclic saturated or unsaturated non-aromatic heterocyclic radical having 5 or 6 members in each ring and wherein the heteroatom or heteroatoms are selected from the group consisting of 1 to 4 oxygen, nitrogen or sulfur atoms and the alkyl moieties linked to the heterocyclic moieties have 1 to 6 carbon atoms or ^{R 3} and R ⁴ with ^p Ojen ^E taken together with the atom ^to choking with at ^kt era ^{l D} EN ESPon is connected in Z 9 may form a five or six membered nitrogen containing heterocyclic ring, R is

18 given for R in addition to the hydrogen atom or R and R together form an alkylidene of 2 to 10 carbon atoms or an alkylidene of 2 to 10 carbon atoms substituted with hydroxyl,

R ⁵ is substituted or unsubstituted alkyl, alkenyl and alkynyl of up to 10 carbon atoms, cycloalkyl and cycloalkylalkyl of 3 to 6 carbon atoms in the cycloalkyl ring and 1 to 6 carbon atoms in the alkyl moieties, phenyl, arylalkyl, arylalkenyl and arylalkinyl wherein the aryl moiety is phenyl and the aliphatic moiety has 1 to 6 carbon atoms, heteroaryl, heteroarylalkyl, heterocyclyl and heterocyclylalkyl, wherein the heteroaryl is a mono-, di- or polycyclic aromatic heterocyclic radical having 5 or 6 members in individual rings and the heterocyclyl is mono-, di- or a polycyclic saturated or unsaturated non-aromatic heterocyclic radical having 5 or 6 members in each ring and wherein the heteroatom or heteroatoms in the above heterocyclic moieties are selected from the group consisting of 1 to 4 oxygen, nitrogen or sulfur atoms and the alkyl moieties associated with said heterocyclic moieties 6 carbon atoms, wherein the abovementioned R 5 radicals are optional substituted with 1 to 3 substituents independently selected from the group consisting of C 1-6 alkyl optionally substituted with amino, fluoro, chloro, carboxyl, hydroxyl or carbamoyl, fluoro, chloro or bromo

STEED

-OCO ₂ R ³

-OCOR ³

4

-OCONR'R li 9

-0 ° -R ^yo

-oxo

4 -NR ³ R

R ^J CONR 34

-NR CO ^R

4

-NR ³ CONR ³ R ⁴ '· 9

-1'IR ³ SR ^J

II o

O

O o \ 9

-Б-К

-SO ₃ R ³

-What _? R ³

--CON ³ R ⁴ • CN 'or phenyl optionally substituted by 1 to 3 carbon atoms, -OR ³ , -NR ³ R ⁴ , -SO ³ R 5, "eritech R" as defined above, fluorine, chlorine ,

CO ₂ R ³ or -CONR ³ R ⁴ , brpniu, alkyl of 1 to 6

4 9 who h, R and R in substit.n or ·· can be attached to

in the other side of the ring so as to form a fused heterocyclic or heteroaromatic ring which may contain further heteroatoms selected from oxygen, nitrogen and sulfur atom is in the ^D or ^{the U-Subs-tuovaný} and I carry ^{B tit} formula --NR and a ^{plurality of projections} l ^alk ene ^yl aa ^{LKI N-yl, Z 1} 0 alkyl, cycloalkyl, cycloalkylalkyl, and alkylcycloalkyl having 3-6 carbon atoms in the cycloalkyl ring and 1-6 carbon atoms in the alkyl moieties, spirocycloalkyl having 3-6 carbon atoms, phenyl, , arylalkyl, arylalkenyl and arylalkinyl wherein the aryl moiety is phenyl and the aliphatic moiety has up to 6 carbon atoms, heteroaryl, heteroarylalkyl, heterocyclyl and heterocyclylalkyl, wherein heteroaryl is a mono-, di- or polycyclic aromatic heterocyclic radical having 5 or 6 members in individual rings and heterocyclyl is a mono-, di- or polycyclic saturated or unsaturated non-aromatic heterocyclic radical having 5 or 6 members in individual rings and wherein the heteroatom or heter the oatoms in the above-mentioned heterocyclic moieties are selected from the group consisting of 1 to 4 oxygen, nitrogen or sulfur atoms, and the alkyl moieties attached to these heterocyclic moieties have 1 to 6 carbon atoms, wherein the substituent or substituents of the above radicals are selected from the group consisting of amino , mono-, di- and trialkylamino, hydroxyl, alkoxy, mercapto, alkylthio, phenylthio, sulfamoyl, amidino, guanidino, nitro, chloro, bromo, fluoro, cyano and carboxyl, and wherein the alkyl moieties in the above substituents have 1 to 6 atoms carbon,

A is a straight or branched alkylene having 1 to 6 carbon atoms,

R is a hydrogen atom, an anionic charge, or a conventional, easily cleavable carboxyl protecting group selected from benzhydryl, allyl, p-nitrobenzyl, 2-naphthylmethyl, benzyl, trichloroethyl, trialkylsilyl wherein alkyls have 1 to 4 carbon atoms, phenacyl, p- methoxybenzyl, acetonyl, o-nitrobenzyl, 4-pyridylmethyl, alkyl of 1 to 6 carbon atoms, pivaloyloxymethyl, acetoxymethyl, phthalidyl, indanyl, methoxymethyl and preferably p-nitrobenzyl, wherein if R ² is a hydrogen atom or a protecting group, an ion is also present and R represents a substituted or unsubstituted mono-, bi- or polycyclic aromatic heterocyclic radical containing at least one nitrogen atom in the ring and generally · 1-4 nitrogen, oxygen or sulfur atom and attached to a through a ring carbon atom and have a ^s c ^s represents ^{Cart for} DUS uv ru ^h ^ ^kt era is ^to var ^t ern ^the pelletized with a ^pi in ^the Nou ^R3 and pharmaceutically acceptable salts thereof, characterized in that

1) reacting an intermediate of formula III

where

8 15

R, R and R are as defined above and

is a conventional readily cleavable carboxyl protecting group selected from the group consisting of benzhydryl, allyl, p-nitrobenzyl, 2-naphthylmethyl, benzyl, trichloroethyl, trialkylsilyl wherein alkyls have 1 to 4 carbon atoms, phenacyl p-methoxybenzyl, acetonyl, o- nitrobenzyl, 4-pyridylmethyl, C 1 -C 4 alkyl, pivaloyloxymethyl, acetoxymethyl, phthalidyl, indanyl, mmethoxymethyl, and preferably p-nitrobenzyl, in an inert organic solvent with a reagent capable of introducing a conventional leaving group L to the position of the intermediate of formula III to form of the intermediate of formula IV

(IV) where

8 15 2 *

R, R, R 'and R are as defined above and L is a conventional leaving group,

(2) the intermediate of formula (IV) is reacted in an inert organic solvent and present base with a mercaptan of formula (A) wherein A is as defined above; and

HS-A

represents a mono-, bi- or u ^- cyclic aromatic, cyclic aromatic radical containing a saturated nitrogen atom in the ring, attached to A via a carbon atom, to form an intermediate of formula II wherein:

R ¹ , R ⁸ , R ¹⁵ , A, R ² and

(II)

It has the meaning given above,

3) reacting the intermediate II with an alkylating agent of the formula II in an inert organic solvent

- X 'wherein ^R5 is as defined above ^{and EA,} X' is a quaternised nitrogen atom in a ring with the common ^and odstuppuící ^{to uρinα,} so that the R ⁵ substituent chink

The product of formula (II) to give a compound of formula (I)

d) wherein R ¹ , R ⁸ , R ¹⁵ , A, R ²

and X 'has the meaning given above, and optionally replaces the anion X' with another carboxyl protecting group R to form the desired unprotected pharmaceutically acceptable salts thereof.

an anion and optionally a compound of the formula I or

The compounds of formula I are potent antibacterial agents for the preparation of these agents.

or intermediates used

The novel compounds of the formula I above contain a carbapenem core

and may be referred to as nines, 1-carba-2-penem-3-carboxylic acid derivatives. to compounds of the formula

Alternatively,

7-oxo-1-azabicyclo (3.2.0) hept-2-ene-2-carboxylic acid. While the protons at the α position are referred to as derivatives, the present invention includes compounds. wherein the relative sterochemistry of α 6 is cis as well as trans, preferred compounds have 5R, 6S (trans) sterochemistry as is thienamycin.

Compounds of formula I may be unsubstituted at the 6-position or substituted by groups

1 as above for the other carbapenem derivatives. More specifically, R can be a hydrogen atom and R can be a hydrogen atom or another substituent or is a hydrogen atom, as disclosed, for example, in European patent application 38 869 (see definition of Rg). Alternatively, R c and r 1 taken together may form C 2 -C 10 alkylidene or C 2 -C 10 alkylidene substituted, for example, with hydroxyl.

Compound ^Y of formula I may be unsubstituted ^which just ^{é p} oloze ^{1 (R 8} = H) or Subbu ^ tuované proviso that both mentioned above for other carbapenem derivatives.

Exact ^R15 may be hydrogen or another subbtituent ^πζ ζ is hydrogen, for example, in published patent at Europ ^{SCE-54917 (even} in a non áefínice ^R1 of ^{R2 b)} No ^b s U.S. Patent 4,350,631.

Preferred proviso that both R ¹⁵ nei other is hydrogen are alkyl having 1 up to 6 carbon atoms, preferably methyl, ^phenyl and ^{-phenylalkyl 1} and ⁶ atoms ^and u ^{h if} uv abylu. ^{With the} ubbtituen R ^15, other than a hydrogen atom may be in either .alpha.- or .beta.-configuration, and it is understood IE předloiený invention includes individual alpha- and beta-soomery even as a mixture thereof .. NeZvýhoOdnjší 1-substituted compounds are those it has a beta-configuration, especially those having a beta-methyl.

have

8 15

Definition ^of R, R and ^R expressions ^y

a) aliphatic alkyl, alkenyl and alkynyl may be unbranched with 1 to 10 carbon atoms, preferably 1 to 6 carbon atoms, preferably 1 and i are part of another substituent, for example cycloalkylalkyl arylalkenyl, then alkyl, alkenyl and alkynyl preferably contain 1 ai carbon ;

or branched carbon atoms, or heteroarylalkyl or

6, preferably 1 to 4, atoms of the chain if

b) heteroaryl includes mono-, bi- and polycyclic aromatic heterocyclic groups having 1 to 4 oxygen, nitrogen or sulfur atoms, preferably 5- or 6-membered heterocyclic rings such as thienyl, furyl, thiadiazolyl, oxadiazolyl, triazolyl, isothiazolyl, thiazolyl, imidazolyl, isoxazolyl, tetrazolyl, oxazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrrolyl, pyrazolyl etc .; .

c) heterocyclyl includes mono-, bi- and polycyclic saturated or unsaturated non-aromatic heterocyclic groups containing from 1 to 4 oxygen, nitrogen or sulfur atoms, preferably 5 or 6 membered heterocyclic rings such as morpholinyl, piperazinyl, piperidyl, pyrrolidolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, pyrrolinyl, pyrrolidinyl and the like;

d) the term halogen includes a chlorine, bromine, fluorine and iodine atom and preferably a chlorine, bromine or fluorine atom.

The term easily removable carboxyl protecting group means a known ester group that is used to protect the carboxyl group during the chemical reaction steps described below and which may optionally be removed by methods that do not result in any destruction of the remaining parts of the molecule, for example by chemical or enzymatic hydrolysis. chemical reducing agents under mild conditions, irradiation with ultraviolet light, or catalytic hydrogenation.

Examples of such ester protecting groups are benzhydryl, allyl, p-nitrobenzyl, 2-naphthylmethyl, benzyl, trichloroethyl, silyl such as trimethylsilyl, phenacyl, p-methoxybenzyl, acetonyl, o-nitrobenzyl, 4-pyridylmethyl and alkyl of 1 to 6 atoms carbon, such as methyl, ethyl or tert-butyl.

Such protecting groups include those that are hydrolyzed under physiological conditions such as pivaloyloxymethyl, acetoxymethyl, phthalidyl, indanyl and methoxymethyl. A particularly preferred carboxyl protecting group is p-nitrobenzyl, which is readily removed by catalytic hydrogenolysis.

The pharmaceutically acceptable salts mentioned above include non-toxic acid salts, for example, mineral acid salts such as hydrochloric acid, hydrobromic acid, hydroiodic acid, phosphoric acid, sulfuric acid and the like, and salts with organic acids such as maleic acid, acetic acid, citric acid. , succinic acid, benzoic acid, tartrate, fumaric acid, mandelic acid, ascorbic acid, lactic acid, gluconic acid and malonic acid. The compounds of the formula I in the form of acid salts can be written as

R = H or a protecting group wherein X is an acid anion. Anion X may be selected such that a suitable salt for therapeutic applications, but in the case of intermediates of the compounds of formula I, anion X may also be a toxic anion.

In this case, the ion can then be removed or substituted with a pharmaceutically acceptable anion to form an active end product for therapeutic use. If present acidic or basic groups in the RR or R ⁵ groups, or in the rest, then the present invention also includes suitable basic or acid salts of these functional groups, e.g. acid addition salts in the case of basic groups and a metal salt (e.g. sodium, potassium, calcium and aluminum), the ammonium salt and salts with non-toxic amines (e.g., trialkyl amines,, procaine, dibenzylamine, 1-ephenamine, N-benzyl-beta-phenylethylamine, N, N'-dibenzylethylenedianiine and the like) in the case of an acidic group.

Compounds of formula I wherein R is a hydrogen atom, an anionic charge, or a physiologically tyrosolyzable ester group together with their pharmaceutically acceptable salts are useful as an internal reagent. The remaining compounds of formula (I) are valuable intermediates which can be converted to the aforementioned biologically active compounds.

G

Preferred compounds of the present invention are compounds of formula I wherein R is hydrogen and r 1 is hydrogen, CH 2 Cl 2 -,

CH., ^X CH-, CH-. OH ³ > no CV CH ₃ ^X OH or | CH3CH -

Of this di-group, the starting materials are ^y where ^R is

OH I _CH3 -chnejvýhodnějšími compounds are those in which m ^ i absolute konnigiuraci 5R, 6S, 8R.

Other preferred compounds of the present invention are those of formula (I) wherein R and R together form an alkylidene radical of formula

BOY

CH

The alkylene moiety (i.e. subetitunet A) in the compound of formula I may have a straight or branched chain and may contain 1 to 6 carbon atoms. Preferred compounds are those wherein A is -C 1-6 wherein n is 1 or 2, and a particularly preferred embodiment includes those compounds wherein A is -C 1-6.

The alkylee moiety of A is attached via a ring carbon atom and an N-substituted quaternary-bonded aromatic heterocycle of formula

C? R ⁵ subetitunet is above ^h o ^d ou ^straight IPA ^and e ^of substituted Al-yl having ^{1 up to 6} carbon atoms, 'and ^ eny ¹ 2 Al 10 carbon atoms alkizyl 2 Al 10 carbon atoms, cycloalkyl of (3-6C) cycloalkylalkyl having 6-6 cycloalkyl and 1-6 carbon atoms in the alkyl moiety; phen (1-6) phenylalkyl having 1 to 6 carbon atoms in the alkyl moiety; (C 2 -C 6) alkenyl moiety, C 2 -C 6 C 1-6 alkyl-C 1-6 alkyl, C 1 -C 6 alkyl-C 1-6 alkyl, C 1 -C 6 -alkyl, C 1 -C 6 alkyl, heterocycles; or a tetrocyclylalkyl wherein the alkyl moiety has 1 to 6 carbon atoms.

ИеЪегоагу! (or the tetrolryl portion of tetrrollyl alkyl) such as R 5 may be a mono-, bi-, or polycyclic, aromatic heterocyclic group containing from 1 to 4 oxygen, nitrogen or sulfur atoms, preferably a five- or four-membered tetnrocyclic ring, such as a ϊϊ 4ζζΙ. furyl, thiilyolyl 'oxa-Hinzolyl, trilyolyl' isothiol, ttilyolyl 'imidazolyl, isozazolyl, benzazilil, oxazooyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl' pyrrolyl and pyrazolyl.

i | Eterocy ^{H k} l ^s l ^(or heterocyclic ^k l ^{i c} and a portion of the heterocyclylalkyl) as R ^{3 with the} substituents of the sub ^can be ^converted by a mono-, bi- or polycyclic saturated or unsaturated non-aromatic heterocyclic group containing 1 to 4 oxygen, nitrogen or sulfur, preferably 5- or 6-membered heterocyclic rings such as morpholinyl, piperazinyl, piperidyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, pyrrolinyl and pyrrolidinyl.

The substituent R may be optionally substituted with 1 to 3 substituents selected independently from the group consisting of

a) alkyl of 1 to 6 carbon atoms, optionally substituted with 1 to 3 substituents such as amino, fluoro, chloro, carboxyl, hydroxyl or carbamoyl;

(b) a fluorine, chlorine or bromine atom;

c) -OR ³ ,

d) -OCO ^ ^3,

e) -OCOR ³ ,

f) -OCONR ³ R ⁴ , clay 9

g) -OS-R \

I »O

(h) -oxo,

(i) -NR ³ R ⁴ ;

(j) R ³ CONR ⁴ -,

k) -NR ⁴ O ^4,

(l) -NR ³ CONR ³ R ⁴

2 9

m) -NR g- ^R ,

O

n) -SR ³

o) -SOR ⁹ ,

O. 9 U ^{p) -} S- ^{RR q) j} -SO ³ R,

r) -CO ³ ,

4

s) -conr ⁴ r ⁴

t) -CN or

u) phenyl optionally substituted with 1 to 3 substituents independently selected from sku3 3 4 pins consisting of fluorine, chlorine, bromine, alkyl of 1 to 6 carbon atoms, -0R, -NR R, -SO ³ R ^j -CC ^g R ^{3 b} of not -CONR ³ R ^{4, w l} e upwards in ^the above pre-designated ^e sulSstiUenHm ^R $ s ^to u ny ^when R ³ and R ⁴ are wearing in nezávkle ^yb wound ^e from the ^{PI to} u n ^y for ^h rnující atom in ^d s ^{to u,} al ^ky l, alkenyl, and alkynyl having 1-10 carbon atoms, cycloalkyl, cycloalkylalkyl, and alkylcycloalkyl having 3-6 carbon atoms in the cycloalkyl moiety and 1 to 6 carbon atoms in the alkyl moiety, phenyl, arylalkyl, arylalkenyl and arylalkinyl wherein the aryl moiety is phenyl and the aliphatic moiety has 1 to 6 carbon atoms, heteroaryl, heteroarylalkyl, heterocyclyl and heterocyclylalkyl, wherein the heteroaryl and heterocyclyl group or moiety is as defined above for R ⁵ and the alkyl moieties attached thereto CA heterocyclic ^s t ^em s ^maj 1 to 6 carbon atoms or R ³ and R ^{4 d} o ^h Roma ^d ysa ^t OMEM ^{d ears} u ^e Itoetomu at least one is attached may form a five or six membered nitrogen containing heterocyclic (as defined above for R) ring and R is as defined above for R ³ except that it is not a hydrogen atom. The most preferred R ⁵ substituent is C 1 -C 6 alkyl, especially methyl.

^{N, and} R VI ^{# 3 in} bstit ^UEN t sj together ^by unique atom in ^the ru ^h in ^{Part I} mo ^ -Butyl

AT·

N21 can form a combined h <terorykl. an organic or heteroaromatic ring, which ring may contain additional, preferably 1 or 2, heteroatoms selected from the group consisting of oxygen, nitrogen and sulfur. For example

Group

can be or

preferably represents a substituted or unsubstituted mono-, bi- or polycyclic aromatic heterocycle containing at least one ring nitrogen atom and 0-5 additional heteroatoms selected from oxygen, nitrogen and sulfur, the heterocyclic ring being attached to A via an atom and has a nitrogen atom in the ring quaternized with the group ^r5 .

Heteroaromatic

the ring may be optionally substituted on available ring carbon atoms with preferably 1 to 5 substituents, preferably 1 to 3 substituents independently selected from the group consisting of alkyl of 1 to 4 carbon atoms, alkyl of 1 to 4 carbon atoms substituted with preferably 1 to 5 carbon atoms 3 hydroxy, amino, C 1 -C 4 alkylamino, C 1 -C 4 dialkylamino in each alkyl, C 1 -C 4 alkoxy, carboxy, halogen (preferably chlorine, bromine, fluorine or iodine atoms), preferably chloro, bromo or fluoro) or sulfo groups, cycloalkyl of 3 to 6 carbon atoms, cycloalkylalkyl of 3 to 6 carbon atoms in the cycloalkyl moiety and 1 to 4 carbon atoms in the alkyl moiety, optionally substituted with 1 to 3 substituents listed above in connection with alkyl (C 1 -C 4), (C 1 -C 4) alkoxy, (C 1 -C 4) alkylthio, amino, (C 1 -C 4) alkylamino C 1 -C 4 dialkylamino, C 1 -C 4 dialkylamino, halogen, C 1 -C 4 alkanoylamino, C 1 -C 4 alkanoyloxy, carboxyl, sulfo, C 1 -C 4 -alkyl-C 1-4 alkyl hydroxy, 0-amido, guamido, phenyl, phenyl substituted with 1 to 3 substituents independently selected from the group consisting of amino, halogen, hydroxyl, trifluoromethyl, (C 1 -C 4) alkyl, (C 1 -C 4) alkoxy, (C 1 -C 4) alkoxy (C 1 -C 4), (C 1 -C 4) dialkylamino, alkyl, carboxy, and sulfo; C 1 -C 4 -phenylalkyl, wherein the phenyl moiety may be optionally substituted with 1 to 3 substituents listed above in relation to phenyl and the alkyl moiety may be optionally substituted with 1 to 3 substituents listed above with respect to C 1-4 alkyl and heteroaryl or heteroarylalkyl, wherein the heteroatom or heteroatoms are selected from the group consisting of 1 to 4 oxygen, sulfur or nitrogen atoms and the alkyl moiety associated with the heteroarylalkyl has 1 to 6 carbon atoms, the heteroaryl and heteroarylalkyl groups being optionally substituted in the heterocyclic ring with 1 to 3 substituents independently selected from the group consisting of hydroxyl, amino, halogen, trifluoromethyl, alkyl of 1 to 4 carbon atoms, alkoxy of 1 to 4 carbon atoms, alkylamino of 1 to 4 carbon atoms, dialkylamino of 1 to 4 carbon atoms in each alkyl , carboxy and sulfo; The compound may be substituted by 1 to 3 substituents selected from the group consisting of hydroxyl, amino, (C 1 -C 4) alkylamino, (C 1 -C 4) alkoxy, carboxyl, halogen and sulfo.

In addition, the ring nitrogen atoms present (in addition to the quaternized nitrogen atom) may be substituted with 1 to 3 substituents independently selected from the group consisting of alkyl of 1 to 4 carbon atoms, alkyl of 1 to 4 carbon atoms substituted with preferably 1 to 3 hydroxy groups, amino groups , (C 1 -C 4) alkylamino, (C 1 -C 4) dialkylamino, (C 1 -C 4) alkoxy, carboxy, halogen or sulfo groups, C 3 -C 6 cycloalkyl, C 3 -C 6 cycloalkylalkyl a (C 1 -C 4) cycloalkyl moiety optionally substituted with 1 to 3 substituents listed above for C 1 -C 4 alkyl, phenyl, phenyl substituted with 1 to 3 substituents independently selected from the group consisting of amino, halogen, hydroxyl, trifluoromethyl, alkyl of 1 to 4 carbon atoms, alkoxy of 1 to 4 carbon atoms, C 1 -C 4 alkylamino, C 1 -C 4 dialkylamino, carboxy and sulfo, C 1 -C 4 phenylalkyl in the alkyl moiety, wherein the phenyl moiety may be optionally substituted with 1 to 3 substituents listed above in relation to phenyl and alkyl the moiety may be optionally substituted with 1 to 3 substituents listed above in connection with a C 1 -C 4 alkyl and heteroaryl or heteroarylalkyl, wherein the heteroatom or heteroatoms are selected from the group consisting of C 1 -C 4 atoms, and the alkyl moiety associated with heteroarylalkyl has 1 to 6 carbon atoms, the heteroaryl and heteroarylalkyl groups being optionally substituted in the heterocyclic ring with 1 to 3 substituents independently selected from the group consisting of hydroxyl, amino, halogen, trifluoromethyl, (C 1 -C 4) alkyl, (C 1 -C 4) alkoxy C1-4 alkylamino; and the alkyl moiety is substituted with 1 to 3 substituents selected from the group consisting of hydroxyl, amino, (C 1 -C 4) alkylamino, (C 1 -C 4) dialkylamino. alkyl, C 1 -C 4 alkoxy in each alkyl, C 1 -C 4 alkoxy, carboxyl, halogen, and sulfo group.

The most preferred substituent on the carbon atom and the nitrogen atom in the ring is alkyl of 1 to 6 carbon atoms, especially methyl.

A preferred embodiment of the present invention described above includes preferred compounds which are those wherein A is - (Cl ₁ -, where n is 1 or 2, most preferably wherein A is -CH ₂ - and wherein

and) 1 8 R and R together form BOY" Cl 2 Z 2 or (b) R ⁸ is hydrogen and ^R is hydrogen, CHo CH-OH l, or CH- C- of ^CH Z ^{c, l} j ^z

4 11 6 7

Particularly preferred are compounds wherein ^{R 1} is hydrogen and K 2 is

OH CN ₃ 4nzejména compounds having the absolute configuration 5R, 6S, 8R.

In a preferred embodiment, the group

represents an aromatic five- or six-membered nitrogen-containing heterocyclic ring containing from 0 to 3 additional heteroatoms selected from oxygen, sulfur or nitrogen.

This aromatic heterocycle may, where possible, be linked to another ring, which may be an unsaturated or saturated carbocyclic ring, preferably a carbocyclic ring of 4 to 7 carbon atoms, an aromatic carbocyclic ring preferably of phenyl, a 4-7 membered heterocyclic ring ( saturated or unsaturated) containing 1 to 3 heteroatoms selected from the group consisting of oxygen, sulfur, nitrogen or NR ^, where R ^ is hydrogen, alkyl of 1 to 6 carbon atoms, optionally substituted with 1 to 2 substituents independently selected from one another 3 _{Is selected} from the group consisting of -OR ', -NRR, -CO _? R, oxo, phenyl, fluoro, chloro,

Bromo, -SO 2 R and -CONR R or phenyl optionally substituted with 1 to 3 substituents ³ _in 33 4 independently selected from the group consisting of alkyl of 1 to 6 carbon atoms, OR, -NR R,

3,334,313 fluorine, chlorine, bromine, -SO-R, -CO-R and -CONR R, where R and R in these R substituents are as defined in connection with R or a 5- or 6-membered heterocyclic ring containing 1 to 3 heteroatoms selected from the group consisting of oxygen, sulfur, nitrogen or NR, wherein R is as defined above.

A 5- or 6-membered aromatic quaternized ring or, where appropriate, a carbocyclic, heterocyclic or heteroaromatic ring attached thereto, or both may optionally be substituted on available ring atoms, preferably up to a total of five substituents on the total ring system, wherein the substituents are listed above in connection with the group

In the preferred embodiment described above, preferred compounds are those wherein A is wherein n is 1 or 2, most preferably those wherein A is -CHg and wherein

a) r! and r 8 together form a group or

b) r 8 is hydrogen and R ¹ is hydrogen,

CHgCHg-,

or

OH

AND

CHchZejména preferred are compounds wherein R is a hydrogen atom ^for R ¹ is u

OH

I ch ₃ ch-, especially those compounds having the absolute configuration of 5R, 6S, 8R.

Another preferred embodiment of the present invention includes compounds of Formula I wherein

represents a residue selected from the group consisting of

a) the rest of the formula

R ^{10 kd} e R ^1, RR and ^R² are independently selected from the group consisting of hydrogen, alkyl of 1 to 4 carbon atoms, optionally substituted preferably with one to three hydroxy, alkylamino of 1 to 4 carbon atoms, dialkylamino in which (C 1 -C 4) alkyl, (C 1 -C 4) alkoxy, amino, sulfo, carboxy, or halogen (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), C 3-6 cycloalkyl C 1 -C 4 alkoxy, C 1 -C 4 alkylthio, C 1 -C 4 alkylamino, C 1 -C 4 alkylamino, C 1 -C 4 dialkylamino in each alkyl, halogen atom (chlorine, bromine, fluorine or iodine) (preferably chlorine, fluorine or bromine), (C 1 -C 4) alkanoylamino, (C 1 -C 4) alkanoyloxy, carboxy, -C (C 1 -C 4) alkyl, hydroxyl, amidino, guanidino, phenyl, f, enyl substituted with one, two or three amino groups, halogen atoms (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), hydroxyls, trifluoromethyls, C 1 -C 4 alkyls or C 1 -C 4 alkoxy groups, phenylalkyl C 1-4 alkyl, wherein the phenyl moiety may optionally be substituted with 1 to 3 substituents listed above in relation to phenyl, and the alkyl moiety may be optionally substituted with 1 to 3 substituents listed above in relation to C 1-4 alkyl and heteroaryl and heteroarylalkyl, wherein the heteroatom or heteroatoms in the above heterocyclic moieties are selected from the group consisting of 1 to 4 oxygen, nitrogen or sulfur atoms, the alkyl moiety attached to said heteroalkyl moiety has 1 to 6 carbon atoms, or wherein two of R ⁶ , RR or together can be joined to form a saturated attached carbocyclic ring, aromatic attached the carbocyclic ring, the attached non-aromatic, heterocyclic ring or the attached heteroaromatic ring, the attached ring being optionally substituted with 1 or 2 substituents as defined above

(b) formula remnants

optionally substituted on carbon by one to three substituents independently selected from the group consisting of alkyl of 1 to 4 carbon atoms, alkyl of 1 to 4 carbon atoms, substituted preferably with 1 to 3 hydroxy groups, alkylamino groups of 1 to 4 carbon atoms, sulfo groups, (C 1 -C 4) dialkylamino in each of the alkyls, (C 1 -C 4) alkoxy, amino, carboxy or halogen atoms (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), C 3-6 cycloalkyl C 1 -C 4 alkoxy, C 1 -C 4 alkylthio, amino, C 1 -C 4 alkylamino, C 1 -C 4 dialkylamino In each alkyl, halogen (chlorine, bromine, fluorine or iodine) (preferably chlorine, fluorine or bromine), (C 1 -C 4) alkanoylamino, (C 1 -C 4) alkanoyloxy, carboxyl, -C (C 1 -C 4) -alkyl; alkyl, hydroxyl, amidino,

O guanidino, phenyl, phenyl substituted with one, two or three amino groups, halogen atoms (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), hydroxyls, trifluoromethyls, (C1-C4) alkyls or (C1-C4) alkoxy Wherein the phenyl moiety may be optionally substituted with 1 to 3 substituents listed above in relation to phenyl and the alkyl moiety may optionally be substituted with 1 to 3 substituents listed above for alkyl of 1 to 4 carbon atoms; 4 carbon atoms and heteroaryl or heteroarylalkyl, wherein the heteroatom or heteroatoms in the above-mentioned heterocyclic moieties are selected from the group consisting of 1 to 4 oxygen, nitrogen, or sulfur atoms and the alkyl moiety associated with the heteroarylalkyl moiety has 1 to 6 carbon atoms or is optionally substituted to form a fused carbocyclic heterocyclic or heteroaromatic ring, optionally substituted with R ⁵ -N or 2 substituents defined

R ⁵

R5

optionally substituted on carbon by one to two substituents independently selected from the group consisting of alkyl of 1 to 4 carbon atoms, alkyl of 1 to 4 carbon atoms, substituted preferably with 1 to 3 hydroxy groups, alkylamino groups of 1 to 4 carbon atoms, sulfo groups, dialkylamino groups having 1 to 4 carbon atoms in each alkyl, alkoxy groups having 1 to 4 carbon atoms, amino, carboxy or halogen atoms (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), cycloalkyl of 3 to 6 atoms C 1 -C 4 alkoxy, C 1 -C 4 alkylthio, amino, C 1 -C 4 alkylamino, C 1 -C 4 dialkylamino in each alkyl, halogen (chlorine, bromine, fluorine or iodine) (preferably chlorine, fluorine or bromine), (C 1 -C 4) alkanoylamino, (C 1 -C 4) alkanoyloxy, carboxyl, -CO (C 1 -C 4) -alkyl alkyl, hydroxyl, amidino, guanidino, phenyl, phenyl substituted with one, two or three amino groups, halogen atoms (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), hydroxyls, trifluoromethyls, alkyls of 1 to 2 C 4 -C 4 alkoxy or C 1 -C 4 alkoxy, C 1 -C 4 phenylalkyl, wherein the phenyl moiety may be optionally substituted with the 1 to 3 substituents mentioned above in relation to the phenyl and the alkyl moiety may be optionally substituted with 1 to aryl and alkyl; the heteroarylalkyl moiety has from 1 to 6 carbon atoms or is optionally substituted to form a fused carbocyclic heterocyclic or heteroaromatic ring, optionally substituted with 1 or 2 substituents listed above in connection with a C 1 -C 4 alkyl, and hetero or heteroarylalkyl, wherein the heteroatom or heteroatoms in the above-mentioned heterocyclic moieties are selected from the group consisting of 1-4 oxygen, nitrogen or sulfur atom of the substituents defined above;

(d) formula remnants

R5

N (III) C is a substituent independently selected from carbon, (C 1 -C 4) alkyl, preferably substituted or optionally substituted with (C 1 -C 4) alkyl, (C 1 -C 4) alkylamino, sulfo, dialkylamino C 1 -C 4 alkyl, C 1 -C 4 alkoxy, amino, carboxy or halogen (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), C 3 -C 6 cycloalkyl , (C 1 -C 4) alkoxy, (C 1 -C 4) alkylthio, amino, (C 1 -C 4) alkylamino, (C 1 -C 4) alkylamino group, halogen (chlorine, bromine, fluorine or iodine), preferably chlorine, fluorine or bromine), an alkanoylamino group having 1 to -CO-alkyl having 1 to 4 li

O-substituted phenyl with fluorine or iodine, with 1 to 4 carbon atoms or alkoxy groups with 1 to 4 carbon atoms, phenylalkyl with 1 to 4 carbon atoms, wherein the phenyl moiety may be optionally substituted with 1 to 3 substituents listed above in relation to phenyl and the alkyl moiety it may be optionally substituted with 1 to 3 substituents listed above in connection with alkyl of 1 to 4 carbon atoms, and heteroaryl or heteroarylalkyl, wherein the heteroatom or heteroatoms in the aforementioned selected from the group consisting of 1 to 4 oxygen atoms; 6 carbon atoms, carbon atoms, C 1-4 alkanoyloxy, alkyl carbon, hydroxyl, amidino, guanidino, phenyl, one, two or three amino groups, halogen atoms (chlorine, bromine, preferably chlorine, fluorine or fluorine); bromo), hydroxyls, trifluoromethyls, alkyls of the heterocyclic moieties are or sulfur and the alkyl moiety

(e) remnants of formulas

or

-R ⁵ wherein X is oxygen, sulfur or NR, wherein R is C 1 -C 4 alkyl, C 1 -C 4 alkyl substituted with 1 to 3 hydroxy, amino, C 1 -C 4 alkylamino, dialkylamino groups up to 4 carbon atoms in each alkyl, 1 to 4 carbon alkoxy, carboxy, halogen or sulfo groups, 3 to 6 carbon cycloalkyl, 3 to 6 carbon cycloalkylalkyl and 1 to 4 carbon atoms in the alkyl moieties optionally substituted with 1 to 3 substituents listed above in connection with alkyl of 1 to 4 carbon atoms, phenyl, phenyl substituted with 1 to 3 substituents independently selected from the group consisting of amino, athalogen, hydroxyl, trifluoromethyl, alkyl of 1 to 4 carbon atoms, C 1 -C 4 alkoxy, C 1 -C 4 alkylamino, C 1 -C 4 dialkylamino in each of the alkyls, carboxyl and sulfo, phenyl alkyl of 1 to 4 carbon atoms in the alkyl moiety wherein the phenyl may be optionally substituted with 1 to 3 substituents listed above in relation to phenyl and the alkyl moiety may be optionally substituted with 1 to 3 substituents listed above with respect to alkyl of 1 to 4 carbon atoms and heteroaryl and heteroarylalkyl, wherein the heteroatom or heteroatoms are selected from the group consisting of 1 to 4 oxygen, sulfur or nitrogen atoms and the alkyl moiety attached to the heteroarylalkyl has 1 to 6 carbon atoms, wherein the heteroaryl and heteroarylalkyl groups are optionally substituted in the heterocyclic ring 1 to 3 substituents independently selected from the group consisting of hydroxyl, amino, halogen, trifluoromethyl, alkyl of 1 to 4 carbon atoms, alkoxy of 1 to 4 carbon atoms, alkylamino of 1 to 4 carbon atoms, dialkylamino of 1 to 4 carbon atoms in each of the alkyl, carboxy and sulfo groups and in the alkyl moiety 1 to 3 substituents selected from the group consisting of hydroxyl, amino, (C 1 -C 4) alkylamino, (C 1 -C 4) dialkylamino, (C 1 -C 4) alkoxy, carboxyl, halogen and sulfo, said heteroaromatic radical is optionally substituted on a carbon atom by one or more substituents independently selected from the group consisting of alkyl of 1 to 4 carbon atoms, alkyl of 1 to 4 carbon atoms substituted with preferably 1 to 3 hydroxy, amino, alkylamino of 1 to 4 carbon atoms, (C 1 -C 4) dialkylamino in each of the alkyls, (C 1 -C 4) alkoxy, sulfo, carboxy, or halogen (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), C 3-6 cycloalkyl carbon atoms, C 1 -C 4 alkoxy, C 1 -C 4 alkylthio, amino, C 1 -C 4 alkylamino a (C 1 -C 4) dialkylamino group, a halogen atom (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), (C 1 -C 4) alkanoylamino, (C 1 -C 4) alkanoyloxy, carboxy, -CO-alkyl &

C 1-4 alkyl, hydroxyl, amidino, guanidino, phenyl, phenyl substituted with one, two or three substituents such as amino, halogen (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), hydroxyl, trifluoromethyl, C 1 -C 4 alkyl or C 1 -C 4 alkoxy, phenylalkyl, wherein the phenyl moiety may be optionally substituted with 1 to 3 substituents listed above in relation to phenyl and the alkyl moiety may be optionally substituted with 1 to 3 substituents as defined above in connection with an alkyl of 1 to 4 carbon atoms, heteroaryl or heteroarylalkyl, wherein the heteroatom or heteroatoms in the above heterocyclic moieties are selected from the group consisting of 1 to 4 oxygen, nitrogen or sulfur atoms and the alkyl moiety associated with said heteroarylalkyl moiety to 6 carbon atoms or optionally substituted to form a fused carbocyclic ring Heterocyclic or heteroaromatic ring optionally substituted with 1 or 2 substituents as defined above;

(f) formula remnants

X c N - NR ^b - V wherein X is oxygen, sulfur or NR, wherein R is C 1 -C 4 alkyl, C 1 -C 4 alkyl substituted with 1 to 3 hydroxy, amino, C 1 -C 4 alkylamino groups , C 1 -C 4 dialkylamino, C 1 -C 4 alkoxy, carboxy, halogen or sulfo, C 3 -C 6 cycloalkyl, C 3 -C 6 cycloalkylalkyl and C 1 -C 4 cycloalkyl carbon atoms in the alkyl moiety optionally substituted with 1 to 3 substituents listed above in connection with alkyl of 1 to 4 carbon atoms, phenyl, phenyl substituted with 1 to 3 substituents independently selected from the group consisting of amino, halogen, hydroxyl, trifluoromethyl, alkyl C 1 -C 4 alkoxy, C 1 -C 4 alkoxy, C 1 -C 4 alkylamino, C 1 -C 4 dialkylamino in each of the alkyls, carboxyl and sulfos Upin, phenylalkyl of 1 to 4 carbon atoms in the alkyl moiety, wherein phenyl may be optionally substituted with 1 to 3 substituents listed above for phenyl and the alkyl moiety may be optionally substituted with 1 to 3 substituents listed above for alkyl of 1 to 4 carbon atoms and heteroaryl and heteroarylalkyl wherein the heteroatom or heteroatoms are selected from the group consisting of 1 to 4 oxygen, sulfur or nitrogen atoms and the alkyl moiety attached to the heteroarylalkyl has 1 to 6 carbon atoms, wherein the heteroaryl and heteroarylalkyl groups are optionally substituted in the heterocyclic ring 1 up to 3 substituents independently selected from the group consisting of hydroxyl, amino, halogen, trifluoromethyl of 1-4 carbon atoms, alkoxy of 1-4 carbon atoms, alkylamino of 1-4 carbon atoms, dialkylamino of 1-4 carbon atoms in each alkyl, carboxy and sulfo; and alkyl parts of 1 to 3 substituents selected from the group consisting of hydroxyl, amino, (C 1 -C 4) alkylamino, (C 1 -C 4) dialkylamino, (C 1 -C 4) alkoxy, carboxyl, halogen and sulfo, said heteroaromatic; the residue is optionally substituted on a carbon atom by one or more substituents independently selected from the group consisting of alkyl of 1 to 4 carbon atoms, alkyl of 1 to 4 carbon atoms preferably substituted.

up to 3 hydroxy, amino, (C 1 -C 4) alkylamino, (C 1 -C 4) dialkylamino, (C 1 -C 4) alkoxy, sulfo, carboxy or halogen (chlorine, bromine, fluorine or iodine), s preferably chlorine, fluorine or bromine), C 3 -C 6 cycloalkyl, C 1 -C 4 alkoxy, C 1 -C 4 alkylthio, amino, C 1 -C 4 alkylamino, C 1 -C 4 dialkylamino. alkyl, halogen (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), C 1 -C 4 alkanoylamino, C 1 -C 4 alkanoyloxy, hydroxyl, amidino, guanidino, phenyl, phenyl substituted with one, two or three substituents such as amino, halogen (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), hydroxyl, trifluoromethyl, alkyl of 1 and 4 carbon atoms or C 1 -C 4 alkoxy, phenylalkyl, wherein the phenyl moiety may be optionally substituted with 1 to 3 substituents listed above in relation to phenyl and the alkyl moiety may be optionally substituted with 1 to 3 substituents listed above with respect to alkyl of 1 up to 4 carbon atoms, heteroaryl or heteroarylalkyl, wherein the heteroatom or heteroatoms in the above-mentioned heterocyclic moieties are selected from the group consisting of 1 to 4 oxygen, nitrogen or sulfur atoms and the alkyl moiety associated with said heteroarylalkyl moiety has 1 to 6 carbon atoms;

(g) formula remnants

I I

NyN — Rc $ ⁵ -n ^ n

ΝγΝ-R

Ν = ΛΙ

R ⁵ -NyN-R ⁵ -N-N-R or

RγΝ where R is C 1 -C 4 alkyl, C 1 -C 4 alkyl substituted with 1 to 3 hydroxy groups, amino groups, C 1 -C 4 alkylamino groups, C 1 -C 4 dialkylamino groups on each of the carbons, C 1 -C 4 alkoxy groups 4 carbon atoms, carboxy, halogen or sulfo groups, C 3 -C 6 cycloalkyl, C 3 -C 6 cycloalkylalkyl and C 1 -C 4 alkyl, optionally substituted with 1 to 3 substituents listed above in connection with C 1-4 alkyl, phenyl, phenyl substituted with 1-3 substituents independently selected from the group consisting of amino, halogen, hydroxyl, trifluoromethyl, alkyl of 1 to 4 carbon atoms, alkoxy of 1 to 4 carbon atoms, alkylamino of (C 1 -C 4), (C 1 -C 4) dialkylamino in each alkyl, carboxy and sulfo group, (C 1 -C 4) phenylalkyl in the alkyl moiety, wherein the phenyl moiety may be optionally substituted with 1 to 3 substituents listed above in relation to phenyl, and the alkyl moiety may be optionally substituted with 1 to 3 substituents listed above with respect to C 1-4 alkyl, heteroaryl and heteroarylalkyl, wherein the heteroatom and heteroatoms are selected from the group consisting of 1 to 4 oxygen, sulfur or nitrogen atoms and the alkyl moiety associated with the heteroarylalkyl has 1 to 6 carbon atoms, said ethereall and heteroarylalkyl groups being optionally substituted in the heterocyclic ring with 1 to 3 substituents independently selected from each other hydroxy, amino, halogen, trifluoromethyl, (C 1 -C 4) alkyl, (C 1 -C 4) alkoxy, (C 1 -C 4) alkylamino, (C 1 -C 4) dialkylamino, alkyl, carboxy and sulfo groups and the alkyl moiety is substituted with 1 to 3 subs substituents independently selected from the group consisting of hydroxyl, amino, C 1 -C 4 alkylamino, C 1 -C 4 dialkylamino, C 1 -C 4 alkoxy, carboxyl, halogen and sulfo.

Su of eluent ^B ST ^{IT y R} and R ³ can be formed when ^{E t s} t ^{d dy} chromium Connections ^{BC H} e ^t EROC ^cli c ^k y not ^b o ^h eteroaromatický ring.

Among the preferred embodiments of the present invention described above, those are those wherein A is - (CH ₂ ) n - and wherein n is 1 or 2, and more preferred are those wherein A is -CH ₂ - and wherein

8

a) R and R together form

BOY

or

1

b) R is hydrogen and R is hydrogen, CH ₂ CH ₂ -

CH

CH-> OH

CHCH or

OH

CH3CH-.

Particularly preferred are compounds wherein R ³ is a hydrogen treatment ^CART ua R ³ is

OH is _3- especially compounds of the absolute configuration 5R, 6S, 8R.

A particularly preferred embodiment of the present invention includes compounds of formula I wherein

represents the rest of the formula

R ^{7 kd} e R ^6, R ⁷ and R ¹⁰ are wearing nezávisl.e having 1 to 4 carbon atoms, alkoxy of 1 to 4 selected from the group consisting of hydrogen, alkyl, alkoxy, carboxyl and carbamoyl, and R3 is as defined above and is preferably alkyl of 1 to 6 carbon atoms, most preferably methyl.

Among the preferred embodiments of the present invention described above, those are those wherein A is - (C 1) - wherein n is 1 or 2, more preferred those wherein A is -C 1 - and wherein

8

a) ^R and R together form a boy ₂ ^{X X} = CH ₃ ^// or

1 b) ^R is hydrogen and ^R is a hydrogen treatment in ^Cart, CH ₃ CH ₂

CH

CH

or

OH

AND

_CH3 CHZejména preferred compounds are

Wherein R is hydrogen and R is

OH

AND

CH3-CH especially those compounds having the absolute configuration of 5R, 6S, 8R.

Another preferred embodiment includes compounds of Formula I wherein

represents the rest of the formula

wherein R ⁵ is LS alk ^y is ^from 1 to 4 carbon atom in ^Y to ^{aluminosilicate,} with the above ^H ou ^{d methylthiophenyl} Ia ^R6 is vocíílcu net> oa ^lkyl having one to four carbon atoms;

b) wherein R ³ is C 1 alkyl having 1 to 4 carbon atoms up to 4 carbon atoms, preferably methyl, and ^R3 are hydrogen atoms or

C)

wherein R ⁵ is or phenylalkyl alkyl up to carbon atoms, carbon atoms, preferably methyl

R is C 1 -C 4 alkyl

(d)

wherein R ⁵ is alkyl, preferably methyl, and alkyl preferably methyl ^R3 is Alk ls ^{y is from 1 to 4} carbon uh ^{l magnesium,}

e) ^kd e R ³ is preferably

f) and ^an alk ^yl methyl or alkyl preferably methyl

R is C 1 -C 4 alkyl,

e ^{kd R5} is alkyl of ^{1 and 4} of ^Y ulilíku, preferably methyl.

From the above-described embodiment of the present invention, preferred compounds are those wherein A is ^(CH 2 ^ n ^"from wherein ⁿ is ^{1 or} 2, preferably those compounds wherein A is _-CH2-, and wherein R ⁸ and ^Y together tvoH noch ₂

X ^{C =}

CH 2 or

b) R ⁸ is hydrogen and R ¹ is hydrogen, CH 3 CH 2

or

OH

AND

CH ₃ CH-.

The invention includes compounds of formula I

1

Particularly preferred are compounds wherein R 0 is hydrogen and R ¹ is

OH

CH3-CH especially those compounds having the absolute configuration of 5R, 6S, 8R.

The most preferred embodiment of the present invention

CH3 or

From this embodiment described above, they are

Preferred are those compounds wherein A is - (CH 2 -) ^- wherein n is 1 or 2, more preferred are those compounds wherein A is - (CH 2 -) and wherein

8

a) R and R together formed

HOCH., '' X 'C =

CH3or

or

ОН

AND

CH3CH-.

Particularly preferred are compounds δ i wherein R is hydrogen and R is

OH l

CH ₃ -CH especially those compounds having the absolute configuration of 5R, 6S,

8R

Particularly preferred compounds of the present invention are compounds of formula

COOR ² wherein R is a hydrogen atom, an anionic charge, or a conventional, easily cleavable protecting group.

carboxyl, provided that when R is a hydrogen atom or a protecting group, it may also be present in the form of an ion and wherein e

G)

-sch ₂

AND

ο)

Ρ)

SCH

where ³ HNMR

3.12 (2H, s ^p e ^kt rum exhibits characteristic peaks ^at Deltim: ^1, 23 ⁽³ ^ ^d, J = ^6, 4 Hz ^ q, J = 4, 8.9 Hz), 3.39 ( 1H, q, J = 2.7, 6.0 Hz), 4.07-4.68 (10H, m), 8.19 (1H, s);

q)

where ³ HNMR

3.15 (2H, (1 H, s));

CH3 (D ₂ O) SPE ^kt rum ^yk azuje characteristic ^peaks oct ^ ^p s ^d and e ^{LT: 23} Ъ (^ ^bd, J = ^6.4 J = 9.0 Hz), 3.37 (1H, qq,

(r)

(t)

in)

or n-ch ₂ coo

The most preferred embodiment of the present invention includes compounds of formula I, wherein it represents

Of the above-described embodiment of the present invention, those compounds wherein A is 18 is preferred n is 1 or 2, more preferred are those wherein A is -OH 3 - and wherein a) R and R together

- (OH 3) r -, where they form

HOOH ' ² \ ^c = /

Or CH 3 '

b) R ⁸ is a hydrogen atom and R ¹ is a hydrogen atom, CH 2 CH 2, ch ₃ ^ CH-, С / CH OH OH \ ‘I C- or CH3CH- Сн / Especially compounds are preferred, 8 1 wherein R is hydrogen and R is OH ch ₃ oh-

especially those compounds having the absolute configuration of 5R, 6S, 8R.

starting materials of formula III may be summarized

Procedure for the preparation of compounds of the formula in the following reaction scheme:

R ⁵ -X '

------>

COOR ²

COOR ²

A variation of the above procedure is apparent

COOR ² optional χ'θ deprotection

----->

from the following reaction scheme

deprotection

When operating as described above, then the starting material of formula III is reacted in an inert organic solvent such as methylene chloride, acetonitrile or ^di meth ^{yl-ORMA i} ds and ⁱ ^ rnún equimolar amount ^{etting IDL} and R c -L ^j as anh ^yd ^ r ^ID oluensu p-sulfonic acid anhydride, p-nitrobenzene sulfonic anhydride, 2,4,6-triítopropylbenzensulfonové acid, methanesulfonic anhydride, trifluoromethanesulfonic acid anhydride, diphenyl chlorophosphate, toluenesulfonyl chloride, p-bromobenzenesulfonyl chloride and the like. wherein L is a corresponding leaving group such as toluenesulfonyloxy, p-nitrobenzene. a group of compounds, diphenoxyphosphinyloxy and other leaving groups which are cleavable by conventional methods known in the literature.

The reaction to remove leaving groups at the 2-position of intermediate III is preferably carried out in the presence of a base such as diisopropylethlamine, triethylamine, 4-dimethylaminopyridine and apoci. ^when te ^pl OTE ^d about -20 ° C ^d of -M0 ° C, preferably ^at a rate ^l OTE about 0 ° C.

in the presence of a base such as diitoproplethylamine, triethylamine, and the like. Intermediate IV may optionally be isolated, but is preferably carried out without isolation or purification steps.

The leaving group L of intermediate IV may also be a halogen atom, in which case this group is cleaved by reacting intermediate III with a halogenating agent such as 03PCl 3 and 03₽Br 2 (φΟΟΡΒ, oxalll chloride and the like) in a solvent such as dichloromethane, acetonitrile, tetrahydrofuran and the like, 4-dimethylaminopyridine is used in the following

Intermediate IV may be reacted with then about to be converted to intermediate II by conventional reaction. Thus, intermediate IV with an equimolar amount of a heteroarylalkyl mercaptan of the general formula

wherein A is a straight or branched alkylene chain having 1 to 6 carbon atoms and

represents a mono-, bi- or polycyclic aromatic quaternizable nitrogen in the ring, the ring in a ring, in an inert organic solvent such as sulfoxide or acetonitrile and in the presence of a base such as sodium, potassium carbonate or 4-dimethylaminopyridine.

the heterocyclic residue containing is attached to A via a carbon atom of dioxane, dimethylformamide, dimethyldiisopropylethylamine, triethylamine, acid carbonate

Temperature for -40 ° C to 25 ° C. to -10 ° C.

cleavage is not critical, but is preferably in the range Nej ^treatment of PE ^to CE REA ^p ch Rovai ^says for ^Quality Control ^í e.g. ^p s ^t e ^{d pl} OTE from about about 0 ° C

The quaternization of the atom results from the reaction of the intermediate (up to about 50 mol%)

II is circulated in the heteroarylalkyl group of the intermediate

II in an inert organic solvent with at least one equivalent of an excess of the alkylating agent of formula

R ⁵ - X k ^d e R ⁵ has the meaning ^d Uve with (chlorine, bromine or iodine, tosylate or triflate.

^TX ea ^χ * is preferably suitable ^{and p} both stages paragraph ^d skupdna atom such as iodine ha ^l ogen) or a part of a sulfonic acid ester such as mesylate

Examples of suitable non-reactive organic solvents are chloroform, methylene chloride, tetrahydrofuran, dioxane, acetone, dimethyl sulfoxide and dimethylformamide. Temperature ^{and Y} 0k 0a ^{N NU} reactions ^and not kňttoká and te ^pl ot ^{y between and} 0 ° ^{C, d} 40 ° ^C are advantageous to total ^Star. Preferably, the reaction step is carried out at room temperature.

Intermediate I 'will have a balancing ion X' (e.g. derived from the alkylating agent used), which can be replaced at this stage or at a later stage, e.g. after the cleavage step, by another ion, e.g. in a conventional manner, for an ion that is more pharmaceutically acceptable. Alternatively, the ion may deprotect during the reaction.

remove performs uses

The degree of cleavage of the protecting group R ~ from the carboxyl group of intermediate I 'with conventional procedures such as solvolysis, chemical reduction or hydrogenation. When a protecting group such as p-nitrobenzyl, benzyl, benzhydryl or 2-naphthylmethyl which can be removed by catalytic hydrogenation, intermediate I 'in a suitable solvent such as dioxane-water-ethanol, tetrahydrofuran-aqueous potassium potassium phosphate, isopropanol or the like is hydrogenated at atmospheric or slightly elevated pressure of 0.1 to 0.4 atmospheres of hydrogen on the hydrogenation catalyst, such as palladium on carbon, palladium hydroxide, oxide salary ^or th like. ^{at room} temp ^ · 0 to ⁵ 0 ° C for about ^{0, 2,} 4 ^{hours and} 4 o'clock.

'.

If group 6 ** is o-nitrobenzyl, then photolysis can be used to remove the protecting group. Protecting groups such as 2,2,2-trichloroethyl can be removed by slight reduction with zinc. The alkyl protecting group may be removed by a catalyst comprising a mixture of a palladium compound and triphenylphosphine in an aprotic solvent such as tetrahydrofuran, diethyl ether or methylene chloride. Similarly, other conventional carboxyl protecting groups may be removed by methods commonly known in the art. Finally, as noted above, compounds of formula I wherein R is a physiologically hydrolyzable ester such as acetoxymethyl, thalidyl, indanyl, pivaloyloxymethyl, methoxymethyl and the like can be applied directly to the host without deprotection, as these esters are hydrolyzable in vivo under physiological conditions .

Rozunu to ^Z e where ^wh e R f, R ^8, R ⁸ or R ⁸ are ^b donating substituents or a heterocyclic ^cli c ^k ý floe ^h is connected to the A substituent comprising a functional group which might interfere with the intended course of reaction, then this group may be protected with a conventional protecting group, and then removed to recover the desired functional group. Suitable protecting groups and procedures for introducing and removing such groups are well known in the literature.

In a variant of the above process, the carboxyl protecting group of intermediate II can be removed prior to the quaternization step. Thus, the carboxyl protecting group is removed as described above to give the corresponding free carboxylic acid, and the free acid is then quaternized with an alkylating agent R-X 'to give the desired quaternized product of Formula I.

If the intermediate Ha without the protecting group is quaternized, water or a non-reactive organic solvent or a mixture thereof can be used as the solvent. Examples of suitable solvents are water, organic solvents such as chloroform, methylene chloride, tetrahydrofuran, dioxane, acetone, dimethylsulfoxide and dimethylformamide and water-organic solvent mixtures such as water-acetone or water-dimethylformamide.

The temperature for the quaternization of intermediate IIa is not critical and temperatures in the range of from about -4 ° C éo about te ^pl Ota m ^ NOSTI ^{without some} moto ^{Gd p} ou t. The advantage S ou ^{d-response p} s ^p Rovai ^says about tepdotě ^{Low: 14} ° C.

If intermediate IIIa is obtained as a carboxylic acid salt, it is necessary to add a strong acid, such as toluenesulfonic acid, to liberate the free carboxylic acid prior to quaternization. It has been found that this greatly facilitates preferential quaternization at the ring nitrogen atom.

The above-described variant is particularly useful when the carboxyl protecting group is more readily removable from the non-quaternized intermediate II than from the quaternized intermediate247 of product I. For example, in the (R) preparation of the product of formula

CH ₃

from the intermediate of formula (R)

removal of the allyl protecting group prior to quaternization results in better yields of the desired end product.

While the above process is suitable for the preparation of the compounds of the present invention, a colleague of Pierre Dextraze has found a new process for the preparation of the compounds of Formula I. This alternative process, which is disclosed and claimed in the copending US patent application filed the same day as this application, is described below example.

In an alternative process for the preparation of compounds of formula I, the intermediate of formula IV

(IV) wherein R ¹ , R ⁸ and R ¹⁵ have a carboxyl group and L is a conventional sulfonyloxy group, with a thiol of formula VII *

as defined above, R ² is a conventional readily removable leaving group such as toluenesulfonyloxy, diphenyloxyphosphinyloxy or a halogen atom and the p-nitrobenzene-protecting group is allowed to react

(VII) wherein A and

-О'- ⁵ are defined above and X is an anion, in an inert solvent and in the presence of base to produce a carbapenem product of the formula and "KD ^e R ^1, R ^8, R ^2, A, R ^15,

and X are as defined above, and optionally the carboxyl protecting group R ² is removed to give the corresponding compound of formula I, deprotecting, or a pharmaceutically-acceptable salt farm.

An alternative procedure utilizes the intermediate formula

(IV) in the published European patent general methods of the present application (defined as X in published iodine, benzenesulfonyloxy, methanesulfonyloxy, which, as mentioned above, has been claimed, for example, by applications 38,869 and 54,917 and which may be prepared as described. patent application 38,869), such as chlorine, bromine, p-toluenesulfonyloxy, p-nitrobenzenesulfonyloxy, trifluoromethanesulfonyloxy, diphenoxyphosphinyloxy or di (trichloroethoxy) phosphinyloxy. A preferred leaving group is diphenoxyphosphinyloxy.

The intermediates of formula (IV) are generally formed in situ by reaction of an intermediate of formula (III)

(III)

COOR ² 'have ^the meanings indicated ^above, with a suitable acylating agent R ° L. Preferred is diphenoxyphosphinyloxy may be prepared by reacting ketoester III with a solvent such as methylene chloride, acetonitrile or dimethyl such as diisotriethylamine, 4-dimethylaminopyridine and the like at a temperature of about -20 ° C, preferably about 0 ° C.

where ^{e 1,} R ⁸ , R ¹⁵ and R ^{2 of} intermediate IV, wherein L in an inert organic formamide with about an equimolar amount of diphenylchlorophosphate in the presence of a base, propylethylamine, up to + ⁴⁰ ° (

Intermediate IV may optionally be isolated, but is preferably used as a starting material for an alternative process without isolation or purification.

The carbapenem intermediate IV is reacted with a quaternary aminthiol of formula VII wherein

(VII) is as defined above and X is an anion. The reaction of acetonitrile, acetonitrile-dimethylformamide, acetonitrile-water or acetone is in the presence of an inert solvent such as tetrahydrofuran, tetrahydrofuran-water, base.

The type of base is not critical. Suitable basses

1,8-diazabicyclo (5.4.0) undec-7-ene, 1,5-diazabicyclo (4.3.0) non-5-ene and C 1 -C 4 trialkylamines in each of the alkyls such as triethylamine, tributylamine or tripropylamine.

are sodium hydroxide, diisorpopylethylamine,

Reaction of intermediate IV and thiol VII may be carried out from - ¹⁵ ° ^C to ^{T t} y EPLO místoosto, preferably in ^N if ^{p s} rovád ⁺¹⁵ ° ^C: Records ^the PE ⁰ ° C.

in a wide range of temperatures-for example ^at rt te ^{pl of} between about -15 ° C

Kar ^b and ^p enemové product of ^p rav (- ^and RCA ^{to a} C kvarternůfo anúnthio ^ VII with intermediate IV are associated with Anic ^ -ntin (e.g., (C / Hj / O) Cl- or the anion associated with the quaternary thiolern) which at this stage may be replaced by another antontum, for example one that is more pharmaceutically acceptable.

This replacement is carried out in a conventional manner. Alternatively, the anion can be removed during the next deprotection step. If the quaternized carbapenem compound and the anion form an insoluble product, then this product can be crystallized and collected by pure filtration.

After the formation of the desired carbapenem product, the carboxyl protecting group R 'in compound I''may optionally be removed by conventional methods such as solvolysis, chemical reduction or hydrogenation. If a protecting group such as p-nitrobenzyl, benzyl, benzhydryl or 2-naphthylmethyl is used which can be removed by catalytic hydrogenation, then intermediate I 'in a suitable solvent such as dioxane-water-ethanol, tetrahydrofuran-diethyl ether-buffer, tetrahydrofuran-aqueous potassium potassium phosphate-isopropanol, or the like. can be reacted under a hydrogen pressure of from 0.1 to 0.4 MPa in the presence of a hydrogenation catalyst such as palladium on carbon, palladium hydroxide, platinum oxide or the like. at a temperature of ^d 0 ^{d 50} ° ^{C when} OBU ^0.24 to ⁴ ho ^di n.

'

If R is a group such as o-nitrobenzyl, photolysis can also be used to remove the protecting group. Protecting groups such as 2,2,2-trichloroethyl can be removed by slight reduction with zinc.

The allyl protecting group may be removed using a catalyst comprising a mixture of a palladium compound and triphenylphosphine in a suitable aprotic solvent such as tetrahydrofuran, methylene chloride or diethyl ether. Similarly, other conventional carboxyl protecting groups may be removed by methods known in the art.

, 2 '

Finally, as mentioned above, compounds of formula I wherein R is a physiologically hydrolyzable ester such as acetoxymethyl, phthalidyl, indanyl, pivaloyloxymethyl, methoxymethyl and the like can be applied directly to the host without deprotection, as these esters are hydrolyzable in vivo under physiological conditions .

The thiol intermediates of formula VII can be prepared, for example, from the corresponding thioacetate compounds of the formula

wherein A is as defined above and

represents a mono-, bi- or polycyclic aromatic heterocyclic radical containing a quaternizable ring nitrogen atom, which ring is attached to A through a ring carbon atom. The thioacetate compound is quaternized by reaction in an inert organic solvent such as diethyl ether, dichloromethane, methylene chloride, dioxane, benzene, xylene, toluene, or mixtures thereof with a suitable alkylating agent of formula

Where R is as defined above and x is a common leaving group such as a halogen atom (chlorine, bromine or iodine, preferably iodine) or an ester portion of a sulfonate such as mesylate, tosylate or triflate. The temperature for the alkylation reaction is not critical, and temperatures ranging from ^{d to 0} ° C to ^d from ⁴⁰ ° C are preferred.

Prior to reaction with the carbapenem intermediate IV, the quaternized thioacetate compound is subjected to acidic or basic hydrolysis to release the quaternary thiol intermediate VII. This hydrolysis is preferably carried out immediately prior to reaction with the compound of formula IV, so as to minimize the decomposition of the relatively unstable quaternary thiol VII.

By suitably selecting solvents, the reaction from intermediate II to the end product i can be carried out without isolation of the various intermediates, i.e. by the so-called single flask process. An example of such a procedure is illustrated below in Example 22.

As with other beta-lactam antibiotics, the compounds of formula (I) may be converted by known methods into pharmaceutically acceptable salts which, for the purposes of the present invention, are substantially equivalent to compounds which are not in the form of a salt.

For example, a compound of formula I wherein R ² is an anionic charge in a suitable inert solvent may be dissolved and then an equivalent of a pharmaceutically acceptable acid is added.

The desired acid addition salt can be isolated by conventional methods, for example, solvent precipitation, lyophilization, and the like. Where other basic or acidic functional groups are present in the compounds of Formula I, pharmaceutically acceptable base addition salts and acid addition salts may be similarly prepared by known methods. .

It will be understood that certain products within the scope of formula (I) may be formed in the form of optical isomers as well as in the form of epimeric mixtures. All optical isomers and epimeric mixtures are included within the scope of the present invention. For example, if the substituent at the 6-position is hydroxyethyl, then the substituent may be either in the R or S configuration and the resulting isomers as well as the epimeric mixtures are within the scope of the present invention.

A compound of formula I wherein R is hydrogen or an anionic charge or a pharmaceutically acceptable salt thereof may also be converted by conventional methods to the corresponding compound wherein R is a physiologically hydrolyzable ester group or a compound of formula I wherein R is a common 2 protecting group carboxyl, can be converted to the corresponding compound wherein R is hydrogen, an anionic charge, or a physiologically hydrolyzable ester group, or a pharmaceutically acceptable salt thereof.

The novel carbapenem derivatives of formula I wherein R is hydrogen, anionic charge, or a physiologically hydrolyzable carboxyl protecting group, or a pharmaceutically acceptable salt thereof, are effective antibiotics for various gram-positive and gram-negative bacteria and can be used, for example, as feed additives for growth enhancers, as food preservatives, as bactericides for industrial applications, for example, water-soluble dyes and orbital waters in paper mills where they inhibit the growth of harmful bacteria and as disinfectants for decomposition and inhibition of harmful bacteria growth in medical and dental instruments.

However, they are particularly useful in the treatment of infectious diseases in humans and other animals caused by Gram-positive or Gram-negative bacteria.

The pharmaceutically active compounds of the present invention may be used alone or may be formulated into pharmaceutical compositions containing, in addition to the active carbapenem components, a pharmaceutically acceptable carrier or diluent.

The compounds may be administered in various ways, with oral, topical or parenteral (e.g. intravenous or intramuscular injection) being the main application. The pharmaceutical compositions may be solid, for example, in the form of tablets, powders, capsules and the like, or liquid, such as solutions, suspensions, or emulsions.

Injectable compositions, a preferred mode of administration, may be prepared in unit dosage form in ampoules or in multi-dose containers and may contain adjuvants such as suspension stabilizing and dispersing agents.

The compositions may be in a form directly applicable for application or in a powder form for reconstitution at a suitable time by the addition of a suitable carrier, such as sterile water.

The dose to be administered depends in particular on the compound used, the particular formulation of the composition, the mode of administration, the type and condition of the patient and the target site, and the organism to be treated. The choice of a particular preferred dosage and route of administration is then left to the skill of the art.

In general, however, the compounds are administered parenterally or orally to a mammal in an amount of from about 5 to 200 mg / kg / day. Administration is generally carried out in divided doses, i.e. three to four times a day.

To elucidate the effective broad spectrum antibacterial activity of the carbapenems of the invention, both in vitro and in vivo and the low toxicity of the compounds, the biological data presented below is compared with the preferred carbapenem compounds of the present invention.

In vitro activity

Samples of the carbapenem compounds prepared in Examples 1 and 2, when dissolved in water and diluted with nutrient medium, exhibited the following minimum inhibitory concentrations (MIC) of µug / ml on said microorganisms. The assay was determined by incubation overnight at 37 ° C by dilution in a tube. N-formimidoylthienamycin was used as reference compound.

In vitro antibacterial activity of the carbapenem derivative according to Example 1

organism MIC (phag / ml) new compound N-formimidoylthienamycin S. Pneumoniae A-9585 0.25 0.004 S. pyogenes A-9604 0.06 0.001 S. aureus A-9537 0.13 0.004 S. aureus + + 50% serum A-9537 0.03 0.016 S. aureus (Pen-Res.) A-9606 0.06 0.008 S. aureus (Meth-res.) A15097 4 0.5 S. faecalis A20688 0.5 0.5 E. coli (10 ~ ⁴ dil.) A15119 0.06 0.016 E. coli (10 ^-3 ) A15119 - 0.03 E. coli ¹ (10 ' ² ) A15119 - 0.06 E. coli (10 ^-4 ) A20341-1 0.03 0.03 E. coli (10 * ³ ) A20341-1 - 0.03 E. coli (10 ^-2 ) A20341-1 - 0.13 K. pneumoniae A-9664 0.25 0.13 K. pneumoniae A20468 0.25 0.06 P. mirabilis A-9900 0.13 0.06 P. vulgaris A21559 0.13 0.03 P. morganii A15153 0.13 0.13 P. rettgeri A22424 0.25 0.25 S. marcescens A20019 0.13 0.03 E. cloacae A-9569 0.13 0.06 E. cloacae A-9656 0.13 0.06

continuation table organism. MIC (µg / ml) new compound

N-formimidoylthienamycin

P. aeruginosa A-9843A 4 1 P. aeruginosa A21213 1 0.25 H. influenzae A-9833 16 16 H. influenzae A20178 32 32 H. influenzae A21522 8 32 B. fragilis A22862 0.03 0.016 B. fragilis A22053 0.03 0.06 B. fragilis A22696 0.25 0.13 B. fragilis A22863 0.03 1

In vitro antibacterial activity of the carbapenem derivative according to Example 2 organism

MIC ( _from µg / ml) new compound

N-formimidoylthienamycin

S. pneumoniae A-9585 0.001 0,002 S.pyogenes A-9604 0.001 0,002 S. aureus A-9537 0.004 0.004 S. aureus A-9537 0.016 0.016 + 50% serum S. aureus A-9606 0.008 0.008

(Pen-Res.)

WITH. ((* aureus et ^J res.) A15097 8 4 with. faecalis A20688 0.25 0.5 E. coli A15119 0.016 0.016 E. coli A20341-1 0.016 0.03 TO. pneumoniae A-9664 0.06 0.06 P. mirabilis A-9900 0.03 0.06 P. vulgaris A21559 0.016 0.03 P. morganii A15153 0.06 0.13 P. rettgeri A22424 0.13 0.13 with. marcescens A20019 0.03 0.03 E. cloacae 19659 0.13 0.06 E. cloacae A9656 0.13 0.06 P. aeruginosa A9843A 8 1 P. aeruginosa A21213 2 0.25

In vivo activity

The in vivo therapeutic effect of the compound of Example 1 and N-fomimidoylthienamycin after intramuscular administration to mice experimentally infected with various organisms is shown in the following table. PD ₅ q values are as follows:

(dose in mg / kg required for 50% protection of infected mice)

Protective effect in intramuscular treatment of infected mice

PD ^ q (treatment (mg / kg) organism infection compound N-formimidoyl- (number according to thienamycin organisms) kladu) P. mirabilis A-9900 3.6 x 10 ⁶ 3.3 3 ^х / 15 ^м P. aeroginosa A-9843a 5.5 x 10 ⁴ 0.3 0,5 ^х P. aeruginosa A-20481 5.4 x 10 ⁴ 0.63 0,4 ^х P. aeruginosa A-20599 1.4 x 10 ⁵ 0.7 0.18 ^X WITH. aureus A-9606 6.6 x 10 ^-8 0.09 0,07 ^х WITH. faecalis A-20688 2.3 x 10 ⁸ 3.3 2.8 ^Х E. coli A-15119 6.2 x 10 ⁶ 0.6 2,5 ^х TO. pneumoniae A-9964 5.1 x 10 ⁶ 2.5 2,2 ^х

* Historical data Sequence of treatment: Except for E. coli A15119 and K. pneumoniae A9964, mice are treated i.m. drug 0 and 2 hours after infection. For E. coli and K. pneumoniae treatment was 1 and 3.5 hours after infection. 5 mice per dose were used for each test.

Toxicity

The toxicity of the compound of Example 1 was determined by intracranial administration to mice and the results are summarized in the table below:

Toxicity for intracranial administration to mice

The highest dose (mg / kg) compound ^ ₅ ο without clinical (mg / kg) signs of toxicity compound according to of Example 1 14 5 N-formimidoyl- thienamycin 32 ~ / 5

^X average 25 / mouse / compound

Blood levels of mice after intramuscular administration

The blood levels and half-life of the compound of Example 1 following intramuscular administration of mg / kg mice are shown in the following table:

Blood level (µg / ml)

compound 10 20 May 30 45 60 90 * tl / 2 ^XX AUC (min) ^ ug.h / ml) compound according to of Example 1 14 10.8 6.8 2.6 0.8 0.6 10 6.3 N-formimidoyl- thienamycin 12.6 9.9 7.3 2.6 0.7 <0.3 9 6 The compounds were dissolve in 0.1M phosphate buffer pH 7. The values are of one test, were used 4 mouse on compound.

tl / 2 - half life in minutes

AUC - refers to the area under the curve.

Urinary excretion

Urinary excretion of the compound of Example 1 after intramuscular administration (20 mg / kg) to mice is shown in the following table:.

Urinary excretion following intramuscular administration of 20 mg / kg to the mouse dose elimination percent

compound 0-3 3-6 hours after 6-24 application 0-24 compound according to of Example 1 26.1 0.5 0.1 26 ^, 7 and 6 ^, 7 N-formimidoul ^_ thienamycin 12.1 0.1 <0.1 12 ^, 2, ^3, 6

Compounds were dissolved in 0.1M phosphate buffer pH 7. Values are from one assay, 4 mice per compound were used.

In vitro activity

Samples of the carbapenem compounds listed below (identified by example numbers) after dissolution in water and dilution with nutrient medium exhibit the following minimum inhibitory concentrations (MIO) of V / Ug / ml on said microorganisms.

Test. ^p rove ^{d i} n en ^b in ^the AC ^and overnight ^{at 37} ° ^{C of} from EA ^E zlkumavce in them. In our eduj ^{L I} C ^I CH sheet is used as the reference compound N-formimidoyl thienamycin.

MIO (uu ^ / ^ ml)

organism Ex 3 4 Ex 5 Ex 6 Ex 7 MK 0787 ^X S.pneumoniae A-9585 0,002 0,002 0.004 0.004 0.004 0,002 S. pyogenes A-9604 0,002 0.001 0.004 0.004 0.004 0,002 S. faecalis A20688 0.5 0.5 0.25 0.5 0.13 0.5 S. aureus A-9537 0.016 0.008 0.004 0.03 0.008 0.004 S. aureus A-9537 0.016 0.03 0.016 0.06 0.03 0.016 + 50% serum S. Aureus A-9606 0.016 0.016 0.008 0.03 0.016 0.008

Pen-res

with. aureus A15097 4 4 8 4 2 4 Meth-res E. coli A15119 0.03 0.016 0.016 0,0 6 0.004 0.016 E. coli A20341-1 0.016 0.016 0.016 0.06 0.004 0.03 TO. pneumoniae A-9664 0.06 0.03 0.03 0.13 0.016 0.06 TO. pneumoniae A20468 0.06 0.03 0.06 0.25 0.016 0.13 E. clocae A-9659 0.13 0.03 0.06 0.25 0.016 0.06 E. clocae A-9656 0.13 0.06 0.13 0.25 0.016 0.06 P. mirabilis A-9900 0.13 0.06 0.03 0,025 0.016 0.06 P. vulgaris A21559 0.016 0.016 0.016 0.06 0.008 0.03 M. morganii A15153 0.13 0.016 0.06 0.13 0.016 0.13 P. rettgeri A22424 0,2 5 0.13 0.13 0.25 0.06 0.13 WITH. marcescens A20019 0.016 0.03 0.06 0.13 0.008 0.03 P. aeruginosa A-9843a 2 2 4 8 1 0.25 P. aeruginosa A21213 0.5 0.13 2 1 0.25 0.25 H. influenzae A-9833 2 2 4 2 = 232 > 32 16 H. influenzae A21518 2 2 4 > 32 > 32 16 B. fragilis A22862 0.25 0.06 0.03 0.03 0.016 0.06 B. fragilis A22696 0.5 0.5 0.25 0.25 0.25 0.13

N- ^f orm ^and Midou ^lt hienamy ^c in

MIC (uc ^ y / ml)

organism Ex 8 Ex 9 eg 10 Ex 11 MK 078 WITH. pneumoniae A-9585 0,002 0.001 0.001 0,002 0,002 WITH. pyogenes A-9604 0,002 0.001 0.001 0,002 0,002 WITH. faecalis A20688 0.5 0.13 0.25 0.5 0.25 WITH. aureus A-9537 0.008 0.004 0.008 0.004 0,002 WITH. aureus A-9537 0.003 0.008 0.06 0.016 0.016

(50% serum)

with. aureus 1-9606 0.03 0.008 0.008 0.016 0.008 Pen-res with. aureus A15097 - - - (meth-res) E. coli A15119 0.016 0.008 0.016 0.016 0.016 E. coli A20341-1 0.03 0.004 0.008 0.03 0.16 TO. pneumoniae A-9664 0.03 0.016 0.06 0.03 0.03 TO. pneumoniae A20468 0.13 0.03 0.13 0.13 0.13 E. cloacae A-9659 0.13 0.03 0.13 0.06 0.13 E. cloacae A-9656 0.06 0.03 0.13 0.13 0.06 P. mirabilis A-990 0.13 0.016 0.13 0.03 0.03 P. vulgaris A21559 0.03 0.008 0.016 0.03 0.016 M. morganii A15153 0.13 0.03 0.13 0.06 0.06 P. rettgeri A22424 0.13 0.06 0.13 0.13 0.13 WITH. marcescens A20019 · 0.06 0.016 0.06 0.06 0.03 P. aeruginosa A-9843A 1 2 32 0.5 1 P. aeruginosa A21213 0.25 0.13 2 0.13 0.13

to N-formimidoylthienamycin

organism> Ex 12 MIC ( _from µg / ml) Ex Ex 14 MK 0787 ^X WITH. pneumoniae A.9585 0,002 0.0005 0.0005 0,002 WITH. pyogenes A-9604 0.004 0.0005 0.0005 0,002 WITH. faecalis A206B8 0.5 0.13 0.13 0.25 WITH. aureus A-9537 0.008 0.008 0.008 0,002 WITH. aureus A-9537 0.016 0.016 0.03 0.008 + 50% serum WITH. aureus A-9606 0.03 0.008 0.016 0.008

Pen-res

with. , aureus A15097 - - - - Meth-res E. . coli A15119 0.016 0.004 0.008 0.016 E. , coli A20341-1 0.008 0.008 0.008 0.016 TO. pneumoniae A-9664 0.03 0.03 0.03 0.03 TO. . pneumoniae A20468 0.06 0.13 0.03 0.06 E. cloacae A-9659 0.06 0.06 0.03 0.06 E. . cloacae A-9656 0.03 0.03. 0.03 0.06 P. . mirabilis A-9900 0.03 0.016 0.016 0.016 P. vulgaris A21559 0.016 0.008 0.016 0.016 M. . morganii A15153 0.06 0.016 0.03 0.06 P. . rettgeri A22424 0.013 0.25 0.06 0.13 WITH. marcescens A20019 0.03 0.016 0.016 0.03 P. . aeruginosa A-9843A 16 32 8 1 P. . aeruginosa A21213 2 2 0.5 0.13

^ -formmdoéthienamycL ·

organism MIC (fg / ml) MK 0787 ^х Ex. 15A Ex. 15B Ex. 15C with. . pneumoniae A-9585 0.0005 0.0005 0.0005 0,002 with. . pyogenes A-9604 0.0005 0.001 0.0003 0,002 with. . faecalis A20688 0.13 0.5 0.5 0.25 WITH. . aureus A-9537 0.03 0.004 0.016 0.004 with. aureus A-9537 0.03 0.016 0.06 0.008 + 50% serum with. . aureus A-9606 0.004 0.008 0,0 3 0.008

Pen-res

with. aureus A15097. - - - - Meth-res E. coli A15119 0.004 0.008 0.06 0.008 E. coli A20341-1 0.004 0.008 0.03 0.016 TO. pneumoniae A-9664 0.008 0.03 0.06 0.03 TO. pneumoniae A20468 0.008 0.016 0.13 0.06 E. cloacae A-9659 0.016 0.016 0.13 0.13 E. cloacae A-9656 0.016 0.03 0.13 0.13 P. mirabilis A-9900 0.008 0.03 0.06 0.06 P. vulgaris A21559 0.008 0.008 0.06 0.016 M. morganii A15153 0.03 0.06 0.25 0.13 P. rettgeri A22424 0.03 0.13 0.25 0.13 with. marcescens A20019 0.008 0.016 0.13 0.016 P. aeruginosa A-9843A 0.5 2 8 0.5 P. aeruginosa A21213 0.03 0.13 0.5 0.13

^{K N-f} om ⁱ m ^id oy ^l t ⁱ n ^hi enamyc

organism ex 16 MIC (Qug / ml) ex 17 KmK 0787 WITH. pneumoniae A-9585 0,002 0.016 0.001 WITH. pyogenes A-9604 0,002 0.016 0.001 WITH. faecalis A20688 1 4 0.25 with. aureus A-9537 0.008 0.25 0.001 WITH. aureus A-9537 0.03 1 0.008 + 50% serum WITH. aureus A-9606 0.016 0.5 0,002

Pen-res

with. aureus A15097 - - - Meth-res E. coli A15119 0.016 0.6 0.008 E. coli A20341-1 0.016 0.6 0.008 TO. pneumoniae A-9664 0.06 0.13 0.03 TO. pneumoniae A20468 0.06 0.5 0.06 E. cloacae A-9659 0.06 2 0.06 E. cloacae A-9656 0.06 2 0.06 P. mirabilis A-9900 0.06 0.13 0.016 P. vulgaris A21559 0.03 0, 13 0.008 M. morganii A15153 0.13 0,5 - 0.06 P. rettgeri A22424 0.25 2 .0.06 WITH. marcescens A20019 0.06 0.13 0.016 P. aeruginosa A-9843A 0.25 ;> 6 3 0.5 P. aeruginosa A21213 0.3 16 0.13

‘N-formimidoylthienamycin

organism Ex 18 WITH. pneumoniae A-9585 1 WITH. , pyogenes A-9604 2 WITH. faecalis A20688 63 with. aureus A-9537 32 WITH. aureus A-9537 _? 63 + 50% serum WITH. aureus A-9606 > 125 Pen-res WITH. aureus A15097 - Meth-res E. coli A15119 16 E. coli A20341-1 16 TO. pneumoniae A-9664 32 TO. pneumoniae A20468 63 E. cloacae A-9659 63 E. cloacae A-9656 125 P. mirabilis A-9900 32 P. vulgaris A21559 32 M. morganii A15153 32 P. rettgeri A22424 32 with. marcescens A20019 32 P. aeruginosa A-9843A 63 P. aeruginosa A21213 63

MIC (pg / ml) ex 19 20 * MK 0787 0,002 0.06 0.001 0,002 0.13 0,002 0/5 16 0.25 0.004 0.5 0,002 0.008 2 0.004 0.016 > 125 0.004 - - - 0.008 1 0.016 0.008 2 0.016 0.03 4 0.03 0.06 4 0.06 0.03 8 0.06 0.03 16 0.06 0.03 4 0.016 0.016 4 0.016 0.06 8 0.06 0.13 8 0.13 0.03 4 0.03 0.5 32 0/5 0.06 16 0.13

^X N-formimidoylthienamycin

In vivo activity

The therapeutic effect of certain compounds of the present invention and N-formimidoylthienamycin (MK 0787) after intramuscular administration to mice experimentally infected with various organisms is shown below.

Values (dose in mg / kg) that provide 50% protection of infected mice are shown.

Protective effect in intramuscular treatment of infected mice

ΡΟ, .θ / 1θδθηί (mg / kg)

Ex 6 Ex 8 Ex 9 Ex 12 Ex 14 Ex 15 (AND) Ex 15 (B) MK 0787 with. aureus A9606 - 0.21 - 0.89 0.07 - - 0.07 E. coli A15119 - 0.86 1,2 - - - - 3 TO. pneumoniae A9664 - 1,8 1, 8 - - - - 3 P. mirabilis A9900 - 1.4 7, 1 - • - - - 9 P. aeruginosa A9843A 0.4 0.19 0, 19 1, 8 0.45 0.39 0.89 1 P. aeruginosa A24081 - 0.33 0.19 - - - - 0.4

Ex 3 ex. 4 Ex 7 MK 0787 WITH. aureus 1 9606 0.07 0.1 0.2 0.07 E. coli A151) 9 1 0.4 0.2 3 TO. pneumoniae A9664 3 3 J 3

continued table

Ex 3 4 Ex 7 MK 0787 P. mirabilis A9900 2 4 2.4 9 P. aeruginosa A9843A 0.5 0.2 0.2 0.5 P. aeruginosa A24081 0.8 0.2 0.1 0.4

Ex 5 MK 0787 S. aureus 0.2 0.07 E. coli 4 2.2 K. pneumoniae 3 2.3 P. mirabilis 10 9 P. aeruginosa A9843A 1.6 0.5

Blood levels of mice after intramuscular administration

Blood levels and half-life of certain compounds therein administered to 20 mg / kg mice are shown below.

of the present invention after intramolecular-

compound C max yug / inl) * T * 1/2 (min) Example 1 14 10 Example 2 13.9 9 Example 3 14.5 10 Example 4 15.5 11 Example 5 - - Example 6 17.7 9 Example 7 19.2 11 Example 8 18.8 11 Example 9 16.7 12 Example 10 20.1 11 Example 11 14.9 11 Example 13 14-8 11 Example 14 15.8 13 Example 15 A 16.7 12 Example 15 B 15.9 10 Example 15 C ” 15.1 10 MK 0787 14.6 10 Example 17 11 a / 8 Example 18 14.9 6 Example 19 27 Mar: 16.7 Example 20 28.4 14

** AUX pig.h / ml)

6.3

5.3

6.9

7.7

8.5

11.8

10.5

8.5

9.5

7.4

6.4

7.6

9.5

7.4

7.3

3.4

3.9

15.1

15.6

Compounds are dissolved in 0.1 M phosphate buffer, pH 7.

Values are from one test, mouse per compound.

^M T ^{l / 2} ~ p ^ ks ^and očias ^Nut c ^h ** AUX - ^pl oc ^H and ^P from ^the KNVB oncentrace in ^the RV ⁱⁿ m times ^s oMotd

The following examples illustrate but do not limit the scope of the present invention.

and

Example 1

Preparation of 1-methyl-4- [2-carboxy-6alpha [1 (R) hydroxyethyl] -7-oxo-1-azabicyclo [3.2.0] hept-2-ene-3-thiomethyl] pyridimium hydroxide inner salt

SCH ₂

CO ₂ PNB co ₂ pnb

Solution 673

SCH ₂

^CH 3 ^I

- pNB = -CH ₂

no ₂

6alpha-1- (R) -hydroxyethyl-3,7-dioxo-1-azabicycloacetonitrile is cooled under a nitrogen atmosphere mmol) of p-nitrobenzyl [2-i] -hept-2-ene-2-carboxylate (1) in 10 ml of a solution of 245 mg (1.90 mmol) of diisopropylethylamine in 1 ml of acetonitrile was added dropwise 510 g (1.90 mmol ·) of diphenylchlorophosphate in 1 ml of acetonitrile.

at -10 ° C. Add in and then in two

The resulting solution was stirred at -10 ° C for 15 minutes to give p-nitrobenzyl-3- (diphenylphosphoryloxy) -α- [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo [3.2.1] m.p. o-hept 2-ene-2-carboxylate.

To this solution was added a solution of 245 mg (1.90 mmol) of diisopropylethylamine in 0.5 mL of acetonitrile, followed by a solution of 270 g (2.16 mmol). Of 4-mercaptomethylpyridine in 0.5 ml of acetonitrile.

The reaction mixture was stirred at -10 ° C for 60 minutes and the resulting white precipitate was filtered off and washed with 5 ml of ice-cold acetonitrile to give 660 mg (76% yield) of compound 2 as white crystals, m.p. 145 ° C.

NMR (DMSO-d6) δ: 1.20 (3H, d, J = 6.0 Hz), 3.2-3.4 (3H, m), 3.7-4.1 (2H, s), 5.05 (1H, d, J = 4.0 Hz), 5.25 (1H, d, J = 14.0 Hz), 5.48 (1H, d, J = 14.0 Hz), 7, 40 (2H, d, J = 5.5Hz), 7.70 (2H, d, J = 8.5Hz), 8.23 (2H, d, J = 8.5Hz) and 8.85 ( 2H, d, J = 5.5 Hz).

IR (KBr) gamma max: 3400, 1790, 1695, and 1600 cm

Anal. Calcd for ^C 22 ^H 21 ^N 3 ° 6 ^S

% C, 58.01;% H, 4.56;% N, 9.23;

Found: C 57.74, H 4.56, N 9.58, S 7.21.

To a solution of Intermediate 2 (660 mg, 1.41 mmol) in acetone (140 mL) was added methyl iodide (5 mL). The reaction solution was stirred at 25 ° C for 8 hours. The solvent was evaporated in vacuo and the resulting light yellow solid was triturated with diethyl ether to give 779 mg (90% yield) of compound 3 as a white amorphous solid, m.p. 130 ° C (dec.).

NMR (DMSO-d 6) δ: 1.15 (3H, d, J = 6.0 Hz), 3.2-3.4 (3H, m), 3.7-4.1 (2H, m), 4.25 (3H, s), 4.30 (2H, s), 5.25 (1H, d, J = 14.0 Hz), 5.50 (1H, d, J = 14.0 Hz), 7.70 (2H, d, J = 9.0 Hz), 8.10 (2H, d, J = 7.0 Hz), 8.25 (2H, d, J = 9.0 Hz) and 8, 90 (2 H, d,

J = 7.0 Hz).

IR (KBr) ymax: 3400, 1770, 1690 and 1640 cm ^1st

H, 4.22; N, 6.82; S, 5.20. Analysis for: C23H24N3O6SI.H2O Calculated: C, 44.39; Found:% C, 44.66;% H, 4.01;% N, 6.84.

To a solution of 779 mg (1.27 mmol) of Compound 3 in a mixture of tetrahydrofuran, water and diethyl ether (80 mL-80 mL-100 mL) was added 140 mg (1.4 mmol) of potassium bicarbonate and 125 mg (0.7 mmol). dibasic potassium phosphate. Then 700 mg of 10% palladium on carbon are added and the mixture is hydrogenated for 45 minutes at 276 kPa on a Paar shaker. The mixture was filtered and the catalyst was then washed with water (2 x 10 mL).

The combined filtrate and washings were extracted with diethyl ether (150 mL) and then a brown powder was obtained by lyophilization. This crude material was purified on C _1Q BONDAPAK reverse phase column (30 g), Waters Associates) and eluted with water at a pressure of 55.2 psi.

Each fraction (20 mL) was separated by high pressure liquid chromatography, and the ultraviolet absorption fractions at _.alpha. = 300 nm were combined and lyophilized to afford 135 mg (32% yield) of compound 4 as a pale yellow solid.

NMR (D 6 O) δ 1.25 (3H, d, J and 2.5 Hz), 3.9-4.2 (2H

J = 6.0 Hz).

m), 4.40

6.0 Hz) (3H, s), 4.72 (2H

2.7-3.2 (2H, m); 3.40 (1H, q, J =

s), 8.10 (2H, d, J = 6.0 Hz),

9.0

8.72 (2H, d i C (KBr) gamma max):

400, 1

755, 1640 ^and 1590

1 cm UV ^lambda max ^(H 2 ⁰⁾

6 nm (epsilon = 7,782)

258 nm (epsilon = 6913).

Λ o

CO2PNB 2

OH

_x / SCH ₂ CH

CO2PNB

pNB = p-nitrobenzyl

A suspension of 1.1 g (2.93 nmol) of diazo compound 1 was purged at room temperature with nitrogen for 5 minutes in 30 ml of anhydrous benzene. 25 mg of rhodium acetate dimer was added and the mixture was heated under reflux for 45 minutes.

The warm solution was diluted with ethyl acetate (25 mL), the catalyst was removed by filtration and evaporated to dryness to give compound 2 as a white solid. This was dissolved in anhydrous acetonitrile ( ²⁰ id) and cooled to ^-10 ° ^C.

To this solution under nitrogen was added diitcprcpylethyOamin (417 mg, 3.2 mmol) and then ⁸¹ 0 mg (3.0 mmol ^l) and rea difenytohtortosííátu mixture ^to CNI ball ^hook 20 m nu ^{i t at} -10 ° C. ^R ea ^CZK n ^e n FRG. Diisorpcpylethylaminem is then treated with (420 mg, 3.2 mmol) and 2- (4-pyridyl) ethanethiol (560 mg, 4.03 mmol) in 2 mL of acetonitrile [J. Org. Chem. 26, 82 (1961) by Ludwig Bauer and Libero A. GardeHa. Jr ^j .

^S śirasā reaction was stirred ^{and for 1} hour ^{at i} nu - ⁵ ° C ^{and -1} ° C 0 _t mettytencMoriiíein then diluted with (100 mL) and successively washed with brine in water (1: 1), 4% H ^ PO 0.5% NaHCO 3, water and brine. *

The organic phase was dried (MgSO 4) and evaporated to give a white solid. This solid was washed with diethyl ether and hexane (1: 1), dried under high vacuum to give 901 mg (63.9%) of compound 3.

^IR ( ^KBR technique) ^{1,790, 1,690} cm ^-1

NMR (CDCl ₃ / DMSO) δ 1.20 (3H, d, J = 3.0 Hz, CH 2), 2.8-3.2 (7H, n), 3.9-4.4 (2H, m), 5.1 (1H, d), 5.4 (2H, g), 7.3 (2H, d), 8.5 (2H, q), 7.76 (2H, d), 8, 3 (2 H, q).

B.

by 3 CO ₂ pNB

The suspension 3 of the carbapenem (890 mg, 1.85 mmol ^), and 7 ml of iodomethane in 200 ml of anhydrous acetone and 1 ^{2 1} ш meth ^ ^ ench ri.du are CH, ^and m ^s from ^{24 h and} n ^{at 25} ° C. FO 18 ^di- N ^and CH reakim the mixture becomes a clear solution. The solvent was removed under reduced pressure and the residue was then washed with diethyl ether to give 920 mg (1.48 mmol) (79.8%) of Compound 4 as a foamy solid.

IR (KBr) 1765, 1690 cm

NMR (DMSO) δ 1.3 (3H, d, J = 3.0 Hz), 3.1 to 3.7 (7H, m), 4.1 (3H, m), 4.3 (3H, s) )

5.38 (2H, d, J = 7.0 Hz), 8.1 (2H, d, J = 3.0 Hz), 8.9 (2H, d, J = 3.0 Hz), 7, Δ (2H, d,

J = 4.0 Hz), 8.2 (2H, d, J = 4.0 Hz).

C.

(H)

To carbapenem 4 (920 mg, 1.47 mmol) dissolved in 90 mL tetrahydrofuran, 90 mL diethyl ether and 90 mL water was added 265 mg (1.51 mmol) dibasic potassium phosphate, 190 mg (1.9 mmol) potassium bicarbonate, and 800 mg of 10% palladium on carbon.

The reaction mixture is hydrogenated at 310 kPa for 1 hour. The catalyst was filtered off through Celite and the filtrate was washed with diethyl ether (3 x, 25 mL). The aqueous layer was lyophilized to give a brownish solid which was purified twice by chromatography on 12 g C _g column and BONDAPAK reverse phase column (Waters Associates) (H ₂ O) to give 55 mg of the fifth

IR-aware (KBr): 1750, 1640 cm ^1st

NMR (D ₂ O) δ 1.30 (3H, d, J = 3.0 Hz), 3.0 to 3.5 (7 µ / m), 4.3) 3H, s), 4.0 to 4 Δ (3H, m), 7.90 (2H, d), 8.70 (2H, d).

Example 3

Preparation of 3- (N-methylpyridin-3-ylmethanthio) -6alpha-1- (R) -hydroxyethyl-7-oxo-1-azabicyclo (3.2.0) hept-2-ene-2-carboxylate

p-Nitrobenzyl 3- (pyridin-3-ylmethanthio) -6alpha- [1- (R) -hydroxyethyl-7-oxo-1-azabicyclo (3.2.0) hept-2-ene-2-carboxylate

To a cooled solution (0 ° C) of 925 * mg (2.66 mmol) of ketone intermediate 5 in 14 mL of acetonitrile was added a solution of 377 mg (2.9 mmol) of diisorpopylethylamine in 1 mL of acetonitrile, giving 786 mg (2.9 mmol) of diphenylchlorophosphate. in 1 ml of acetonitrile under nitrogen.

MMN ^IKL characterized ^extensible by ^k m ^{Ch 15} minutes ^EXAMPLE ° C and PA ^to the ^P / I ^d and solutions. ^{377 g} · m ^(2, 9 mmol) of 3-mercaptomethylpyridine prepared as in Can. J. Chem. 56, 3068 (1978) in ^{2 ml of} acetone. Rea ^{CZK tup} mites ^{hook the} ball nut 90 m ^{and p} s ⁰ ° C.

The precipitate was filtered off and washed with 20 mL of ethyl acetate to give 950 mg (60% yield) of the title compound as white crystals.

NMR (DMSO-d6) δ: 1.30 (3H, d, J = 6.0 Hz), 3.4-4.2 (5H, m), 4.25 (2H, s), 5, 1 (1H, d, J = 4.5Hz), 5.40 (2H, Abq, J = 14.4Hz), 7.2-8.5 (8H, m).

IR (KBr technik.a) imbda ^ ^ * ^ ^{0 3550 1775} and ^{1580 cm 3rd}

^H , 4.65; N, 9.23. S, 7.04.

Found: C, 57.19; H, 5.19; N, 8.76; S, 7.08.

OH

CO ₂ pNB

OH

CO ₂ pNB

CH3

3- (N-methylpyridin-3-yl-methanthio) -6alpha- [1- (R) -hydroxyethyl] ^-7- oxo-1-azabicyclo (3.2.0) heptr-2-ene-2-carboxylate

To a solution of Compound 19 (730 mg, 1.56 mmol) in acetone (120 mL) was added methyl iodide (5 mL) and the reaction mixture was stirred at room temperature for 18 h. The precipitate was filtered off and washed with acetone (10 mL) to give 940 mg (100% yield) of quaternized pyridine 20 as a pale yellow powder.

NMR (DMSO-d 6) δ; 1.25 (3H, d, J = 5.8 Hz), 3.6-4.3 (5H, m), 4.20 (3H, s), 4.25 (2H, s), 5.25 (1H, d, J = 7.2 Hz), 5.40 (2H, Abq, J = 12.16 Hz) and 7.6-9.2 (9H, m).

^Ic ( ^K Br technique) lambda: ^{3 300} ^ ^{1 765} and ^{1 690} cm ³ .

max

Analysis for C ₂ ^H 24 ^N 3 3 ° 6Siíi calculated 46.24% C, 4.05% H, 7.03% N, 5.37% S;

found: 45.82% C, 4.11% H, 6.87% N, 6.10% S.

To a solution of 933 mg (1.6 mmol) of compound 20 in 90 mL of tetrahydrofuran and 90 mL of ether was added 200 mg of KHCO 3 and 349 mg of 4 HPO 4 in 90 mL of water, followed by 1.0 g of palladium on carbon. The mixture is hydrogenated for 45 minutes at 310 kPa on a Paar shaker. The reaction mixture was filtered through celite, the catalyst was washed with water (2 x 10 mL).

The combined filtrate and washings were extracted with ether (2 x 100 mL) and lyophilized

afforded a yellow solid which was purified on a C _1O BONDAPAK one of the phases (8 g), eluting with 5% acetonitrile in water under pressure (kPa Waters.

Associates) column with reverse

Each 15 ml fraction was assayed by high pressure liquid chromatography and fractions ^L amb ^d and max 3 (^ 0 nm with ^p pole ^s and ^Ly of ^ili recrystallisation ^of the Z ^{and KA 230} m ^{g of} the ^DPI dried in a ^f orm ^of seduces ^ylw u ^t ACTIVE crystals, mp 1 ³⁰ ° C with ultraviolet (4 ^3% yield off ^from the e ^k) (rozdad).

absorption at s ^{l No.} ou en ⁱ ny said ^E

NMR (D 6 O) δ 1.25 (3H, d, J = 7.0 Hz), 3.12 (1H, q, J = 7.2 Hz, 1.6 Hz), 3.9-4.6 (3H, m), 4.25 (2H, (2H, dd, J = 7.9Hz, 1.6Hz)), 3.42

s), 4.42 (3H, s) and 8.0-9.0 (4H, m >).

^IR (techn ^{IK K} Br) ama ^g max: ^{3400, 1 75} 0 and ¹⁵⁸⁰

- 1 cm

UV λ max (10 → 0): 298 nm (epsilon = 8058).

Analysis: for ^ ₆ Η) οΝ ₂ ° ₄ 8ι.2Η ₂ ° calculated

% C, 51.87;% H, 5.44.

% C, 51.95;% H, 5.66.

7.56

7.56

N;

N.

Example

3- (N-meth ^yl lp ^y pyridin-2-yl-methane) -6alfa- [l- (R) h ^y droxyeth ^{yl-7-oxo-bicyclo} (32.0) hept-2-ene-2-carboxylate

Preparation

PN ⁱ ine ^b enz ^yl-3- (p ^s r ^s d ^{i n-2-yl} methyl ^H H n ^{t s} o) - ⁶ and ^LF alpha- ^{[ 'l} - (R ^ y ^^ oxyethylene ¹ 7- oxo-1-azabicyclo (3.2.0) hept-2-ene-2-carboxylate

K oc ^hl Azen ^é ^ extended to ⁽⁰ ° ^{C) 925} m ^{g (2,} 65 mmo ^{l) e t} Limit ^IP roducts ⁵ 14 m ¹ acetonúrdu a solution of 377 mg (2.92 mmol) of diisopropylethylamine in 1 ml of acetonitrile and then 786 mg (2.90 mmol) of diphenylchlorophosphate in 1 ml of acetonitrile under nitrogen were added.

MMN ^IKL characterized mites ^k m ^s c ^HA 15 m nu ^{i t} p ^{s 0 C} dšisopropylethylaminu in 1 ml of acetonitrile followed by f ^ ddtou ^[STBY Raven ^eh of the ^dl postopem ^p e n ^C. J. ^C hem. 56 mites are then ^{added to 377} m ^{g (2.92} mmo ^l) se'přidá 350 mg (3.0 mmol) of 2-mercaptomethyl ^{3 068 (1 97} 8) 7 ^{1 1} m acetontordu.

^{R CZK} ea n ^s m ^s solution chá hoddiy ^{2 at -} 10 ° ^C, methylene chloride to give 650 mg (54% yield) of a yellow powder.

the title compounds in form i

NMR (DMSO-d6) δ: 1.26 (3H, d, J = 7.0 Hz), 2.7-3.5 (4H, m), 3.9-4.3 (2H, m), 4.2 (2H, s), 5.42 (2H, Abq, J = 14.4 Hz) and 7.2-8.8 (8H, m).

IR (KBr) gamma max: 3,400.1

Analysis calculated for: ^ε 22 ^Η 21 ^Ν 3 ^Ο 6 ⁸ 1 C 58.01%, H 4.65%, 9.23% found C 57.56%, H 4.92%, 8.94%

775

N,

N, and 1690 cm

7.04% S;

7.03% S.

CH ₃ I acetone

H2

------------ R.

Pd / C

Ιθ

OH

3- (N-methylpyridin-4-yl) -than-6-ol^lfand^-[b-O] -hydroxyethyl-7-oxo-1-azabicyclo (3.2.0)^hE^pt -2-ene-2-carboxylate

To a solution of 650 mg (1.39 mmol) of 22 in 100 mL of acetone was added 4 mL of methyl iodine. The reaction mixture was stirred at room temperature for 3 days. The precipitate was filtered off and washed with acetone (10 mL) to give 500 mg (60% yield) of quaternized pyridine 23 as a light yellow solid.

NMR (DMSO-d6) δ: 1.26 (3H, d, J = 7.0 Hz), 3.9-4.2 (2H, m), 4.4 (3H, s), 4.78 ( 2H, s), 5.2 (1H, d, J = 3.9 Hz), 5.50 (2H, ABq, J = 14 Hz) and 7.8-9.4 (8H, m).

IR (KBr) gamma max: 3400, 1765 and 1690 cm

Analysis calculated for: ^C 23 ^H 24 ^N 3 ^O 6 ^{S 1} H: ^C, 46.23; H, 4.05; N, 7.03; S, 5.37.

found: C 45.62, H 4.27, N 6.80, S 5.30.

To a solution of compound 23 (1.0 g, 1.167 mmol) in tetrahydrofuran (90 mL) and ether (90 mL) was added KHCO3 (215 mg, 2.15 mmol) and K2 PO4 (374 mg, 2.1 mmol) in water (90 mL). 1.0 g of 10% palladium on carbon is added.

The mixture is hydrogenated at 310 kPa for 45 minutes on a Paar shaker. The reaction mixture was filtered through celite and the catalyst was washed with water (2 x 10 mL). The combined filtrate and washings were extracted with ether (2 x 100 mL) and lyophilized to give a yellow solid which was purified on a Co BONDAPAK (Waters Associates) reverse phase column (10 g), eluting with 5% acetonitrile in water at 55 kPa.

Each 15 ml fraction was assayed by high pressure liquid chromatography and the ultraviolet absorption fractions at lambda max 30 nm were pooled and lyophilized to yield

390 mg (44% yield) of the title compound. Recrystallization of this material from water-acetone-ethanol ^to ZIS and ^even fine ^{s h} l is ^cyclic, mp ¹⁹⁴ and ^{from 196} ° C (dec).

se

NMR (.delta. 2 DEG) .delta.: 1.30 (3H, d, J = 6.2 Hz), 3.2 (2H, q, J = 9.0 Hz, 3.6 Hz), 3.46 q, J = 6.0 Hz, 2.7 Hz), 4.1-4.6 (3H, m), 4.60 (3H, s) and 7.9-8.9 (4H, m).

(1H

IR (KBr) gamma max: 3400,

755 and 1590 cm

UV λ max (H 2 O): 292 nm (epsilon = 8092).

Analysis calculated for: ^C 16 ^H 28 ^N 2 ° 4 ^N 1. ^2H 2 ° calculated

% C, 51.87;% H, 5.44;

% C, 51.37;% H, 5.69;% N, 7.37;

OH

Example

P ^pava rava

3- ^(N-methyl-py ridin- ² - ^yl ethanthio) -6a ^LF A- ^£ l- ^(R) - ^{hydroxyacetophenone y} eth ^{yl-7-oxo-bicyclo} (3.2.0) hept-2-ene 2-carboxylate

O

II

C1P (O0) ₂

CO ₂ pNB

P ^^ robcnzyl-3- (pyridin-2- ^{yl lethanthio)} 6a ^lf f 1 (R) hydroxyethyl ^^ - oxo ^ aza ^^^ o (3.2.0) hept-2-ene-2 -carboxylate

To a cooled solution of ketone intermediate 1.78 (5.0 mmol) in 25 mL acetonitrile was added g (5.0 mmol).

710 mg (5.5 mmol) of diisopropylethylamine in diphenylchlorophosphate in 1 ml of acetonitrile in ml of acetonitrile and then a nitrogen atmosphere is added.

1.4 ⁰ ° ^C and ^p Otom mites are ^{added to 710} m ^g of Zn ⁱⁿ the solution characterized ^IKL m c ^{s t h and 20} mine ^at diisopropylethylamine in 1 ml of acetonitrile and then added an aqueous solution of thiol 54 850 fpřipraveného according to J. Org. Chem. 26, 82 (1961)] in 2 ml of acetonitrile.

mg ^(5.5 mmol) of (6.1 mmol) of ^R EAK ^{N NU} is a mixture ^of m ^I C ^{hook 60} minutes. ^{EXAMPLE 0C.} Precipitation from ⁱ to ^{p fil} Truji rom ^{y methylcarbazole} Lorido ^h enc (20 mL) to give 1.3 g (57%) of the title compound as a yellow solid.

AND

NMR (CDCl3): 1.25 (3H, d, J = 6.5 Hz), 2.6-3.4 (7H, m), 4.2-4.6 (2H, m), 5.30 and 5.65 (1H each, ABq, J = 14 Hz) and 7.2-8.5 (8H, m).

IR (KBr) ymax: 3400, 1780 and 1680 cm ^1st

3- (N-methylpyridin-2-ylethanthio) -6alpha-1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo (3.2.0) hept-2-ene-2-carboxylate

To a suspension solution of 800 mg (1.7 mmol) of compound 53 to 50 mL of acetone was added 5 mL of methyl iodide.

The reaction mixture was stirred at room temperature for 48 hours. The precipitate was filtered off and washed with acetonitrile (15 mL) to give 810 mg (76% yield) of quaternized pyridine 55 as a pale yellow powder.

NMR (DMSO-d 6) δ: 1.20 (3H, d, J = 5.6 Hz), 3.2-4.3 (9H, m), 4.20 | 3H, s), 5.26 and 5.55 (1H each, ABq, J = 15 Hz) and 7.8-9.2 (8H, m).

IR (KBr) ymax: 3400, 1770 and 1690 cm ^1st

To a solution of 790 mg (1.27 mmol) of compound 55 in 100 mL of tetrahydrofuran and 100 mL of ether was added 100 mL of pH 7.0 buffer followed by 1.0 g of 10% palladium on carbon. The mixture is hydrogenated at 276 kPa for 40 minutes on a Paar shaker.

The reaction mixture was filtered through celite and the catalyst was washed with water (2 x 10 mL). The combined filtrate and washings were extracted with ether (3 x 100 mL) and lyophilized to give a yellow powder which was purified on a BONDAPAK (Waters Associates) column (30 g), eluting

10% acetonitrile in water at 55 kPa. Each 15 ml fraction was assayed by high pressure liquid chromatography and the fractions having ultraviolet absorption at lambda max 300 nm were combined and lyophilized to afford 65 mg (15% yield) of the product as a yellow powder.

NMR (D ₂ O) δ: 1.25 (3H, d, J = 6.2 Hz), 3.1-3.6 (7H, m), 4.0-4.3 (2H, m), 4.32 (3H, s) and 7.8-8.9 (4H, m). IR (KBr) ymax: 3400, 1750, 1590 cm UV lambda max (H ₂ 0): 300 nm (epsilon = 8108).

lad 6

Preparation of 3- (1-propylpyridin-4-ylmethanthio) -6alpha-1- (R) hydroethyl-7-oxo-1-azabicyclo (3.2.0) hept-2-ene-2-carboxylate

CO3P ^ B

P-nitrobenzy1-3- ^{(pyrifin-4-yl-methane)} -6.alpha.-F1- (R) "hydfrjxycehylJ" ^{7-oxo-1-azabicyclo} (3.2.0) yept-2-ene-2-carboxylate

Solution 67³ mg (^1,8⁶ mmol)^p-nitro^benzyme^y-6al^Fand^{- / -}(R) -^yyfroxyrehy-y-dwxo-i-aya -^biC^yklof ^ .connpt-enen-yartox ^ tu⁵ in¹Cool 0 ml of acetonitrile^-10 ° C in atooo^Fthe era of nitrogen. Add a solution of 245 mg (1.90 mmol) of dlisopropyleyhyaolin in 1 mL of DMF followed by dropwise addition of 510 mg (1.90 mmol) of diphenylphosphoryl phosphate in 1 mL of acetonitrile over 2 min.

Vznnklý Royt ^the ball ^YA P ^{RL -10} ° C for ¹⁵ minutes with číož yistá ^p i ^t -ni o ^b ^ en; _/ ^{R "3} - ^(d Ifen ^{r] f} or ^f os ^yl ox ^{y) -6} alpha- ^{[l ((R)} -h ^yd rox et ^{y y} y ^^ - ^ XO L-ayabicy ^y lo - ⁽³ · ² .0) ^ pt-HN ^2- .artoxylít. to this solution was added a solution of 245 mg (1.90 mnuml) dlisorp ^pyáet-y ylaoinu nitrile and then a solution of 270 mg (2.16 miml ·) 4-Inocapptothiothiopyridium nitrile.

0.5 ml 0.5 ml acetov mixture acetoReakční ball ^Y and Pr ¹ -1.0 ° ^C-60 min and washed with 5 ml ice-lceto ilrláu thereby) Iška 660 mg of 145 ° C.

in the form of a ^white bull ^y ^^ taiu tt

^{characterized} consists ^of en ^{and a white} precipitate (76% yield) of compound 51 odfUtruje

NMR (DMSO-d 6) δ: 1.20 4.25 (2H, s), 5.05) 1H, d, J = 7.40 (2H, d, J = 5.5 Hz), 7.70 J = 5.5 Hz).

(3H, d, J = 6.0 Hz), 3.2-3.4

4.0 Hz), 5.35 (1H, d, J = 14 (2H, d, J = 8.5 Hz), 8.23 (2H, d, (3H,

Hz), m, 3.7-4.1 (2H, m,

5.48 (1H, d, J = 14.0 Hz),

J = 8.5 Hz) and 8.58 (2H, d,

IR (KBr) ymax: 3400, 1790 and 1695 cm ¹

Annals)! for C22H21N3O6C calculated 58.01 C, 4.5b%

% S.

N, 9.58

l

3- (1-allylpyridin-4-ylmethanthio) -6alpha-1- (R) -hydroxyethyl-7-oxo-1-azabicyclo (3.2.0) hept-2-ene-3-2-carboxylate

To a solution of Compound 51 (900 mg, 2.13 mmol) in acetone (150 mL) was added allyl bromide (2 mL) and sodium iodide (380 mg). The reaction mixture was stirred at room temperature for 48 hours and the solvent was removed in vacuo to give a yellow solid. This material was suspended in 120 mL of acetonitrile, filtered and evaporated in vacuo to give 1.0 g (8 &apos; yield) of the title compound as a yellow solid.

NMR (CD, CN) .delta 1.20 (3H, d, J = 6.2 Hz), 3.0-3.4 (4H, m), 4.0-4.4 (4H, m), 5.1-5.6 (4H, m) and 7.4-7.9 ’8H, m). IR (KBr technology) gamma: 3400, 1 max 770 and 1690 cm. Analysis: for C. 1 H 2 6 ^N ° 6 3 1 ^{1 S)} calculated 48.16% C, 4.21; H, 6.74 % N, 5.15% S; climbing 4 8,55% C, 4.46% H, 6.69 % N, , 5.15% S.

Pd / C

- (1-propylpyridin-4-ylmethanthio) -6alpha- [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo (3.2.0) hept-2-en-2-carboxyl;

To a solution of Compound 52 (1.27 g, 2.15 mmol) in 100 mL of tetrahydrofuran and 100 mL of ether was added 100 mL of pH 7.0 buffer, followed by 1.0 g of 10% palladium on carbon. The mixture is hydrogenated at 276 kPa for 40 minutes on a Paar shaker.

The reaction mixture was filtered through celite and the catalyst was washed with water (2 x 10 mL) and filtered. This gives a yellow powder which is purified on a 13ONDAPAK (Waters Associates) column (40 g), eluting with 10% acetonitrile in water at 55 kPa.

Each 5 ml fraction was assayed by high pressure liquid chromatography and the ultraviolet absorption fractions at lambda max 300 nm were combined and lyophilized to give 48 mg (6% yield) of the title compound as a yellow powder.

NMR (D 6 O) δ 0.95 (311, t, J = 7.5 Hz), 1.25 (311, d, J = 7.0 Hz), 2.05 (2H, sextet, J - 7.5 liz), 3.10 (211, dd, L-10 Hz, 2.5 liz), 3.35 (1H, dd, J = 6.5 Hz, 2.5 Hz), 4.0- 4.8 ((2H, m), 7.1 (2H, d, J = 6.0 Hz), 7.1 (2H, d, J = 6.0 Hz).

IR (KBr) technj q ka ima них: 1 41) 0 7 I ^r> OA 1590 honored

Λ na I ysis: for Η ,, 11, N, O S. 2II calculated

% C, 54.52;% H, 6.10

C, 54.32; H, 6.03

Example

3- (N-meth ^{yl-3-meth yl} lp ^y ri ^di n-2-methane ^t h ^{i o) LF} 6a A- ^{£ l} - ^{(R) hyd} rox ^y e ^ethyl] -7-oxo'Příprava

-1- (2-carboxylate) (3.2.0) hept-2-ene-2-carboxylate

2) NaOH

HCl

3-Methyl-2- (1-methyl-thiopyridine thiourea) in 60 ml Ethanol was evaporated and dissolved

A solution of 2.45 g (17.0 mmol) of chloro compound 36 and 1.37 g (18.0 mmol) of absolute ethanol was heated to reflux for 5 hours. and addition of ether gave 3.08 g (72% yield) of the isothiouronium salt, in 10 mL of water containing 1.44 g (26 mmol) of sodium hydroxide.

^The organic solution is ^p Otom per ^hour iv ^{and 5} m ^and nut to ¹⁰⁰ ° C at ⁵ ° C ^{EXAMPLE id} anime ^{KY If} n ^y vinegars ^s pH was adjusted combined ether extracts were washed with 5% aqueous va ^t MOS ^FER e ^d ears ^to u. ^R ea ^CZK ocHaclí the mixture to ^6.4 and the r ex ^{t h} u is the etoerem (4 x 50 mL). solution, sodium bicarbonate and solvent solution gave 1.4 g (83% yield) of sodium chloride. Evaporate the dried (MgSO4) thiol 37 as a yellow oil, which is used for the next step without further purification.

NMR (CDCl3): 2.20 (2H, s), 2.5-2.7 and 6.9-8.2 (3H, m).

(1H, broad s), 3.8 (2H, t, J = 6.5 Hz)

OH

£ ČOjpNB

^p-n^andtro^benžy^l-³- [³~ me^thy^lpyr^idin-²-^ylmet^han^thio] -6a^Fand - [1- U^R) -^hydrox^yet^bullfinch] - / - oKok-azates cyclo (3.2.0) heptn-2-en-2-carbox^ylath^TO ocazene solution (⁰ ° C)^{1.74 g (}5,0 mmo^l) totomez ^ ro ^ Hu⁵ ve²⁵ m^l 960 g (5.8 mmol) of diisopropylethylamine in 2 ml of acetonitrile are added to the acetonitrile, and 1.4 g (5.8 mmol) of diphenylchlorophosphate in 2 ml of acetonitrile are added under a nitrogen atmosphere. The resulting solution was acetonitrile and then 810 mg of mercaptomethylpyridine 37 in 3 mL of acetonitrile. The reaction mixture was stirred for 1 h^andC^{há při} Low: 14 ° C² hod ^ y.^Stražen^andna se o^dfiltruje,^prom^yis acetone^andtrilen ^^{, while}from which^andwith^ká l,⁵⁶ g (6⁰% Výtě^ofE^to) compounds^den^E in on^dpisu in form Шё^pSolids, m.p. 1⁴⁵ Deň: 32 ° C.

NMR (DMSO-d 6) δ: 1.23 (3H, d, J = 6.5 Hz), 2.30 (3H, s), 3.1-4.3 (6H, m), 4.35 ( 2H, s), 5.20 and 4.45 (1H each, ABq, J = 15.0 Hz) and 7.3-8.4 (7H, m).

IR (KBr technique) gamma max:

Analysis; for C24H26 ^N ° 3 9S2F calculated 47.91% C, 4.69% Found 47.72% C, 4.34%

400, 1,767 ^and 1,695 ^cm "\

H, 6.98% N, 10.66% S;

H, 6.72; N, 11.22.

FSO ₃ CH ₃

->

Pd / C

3- 0 ^ - 'b' / li ^ 't' al^f1 (JR) -hydroxy-thi-oxo-1-azate (3.2.0) hept-2-ene-2-carboxylate

To a solution of 680 mg (1.45 mmol) of compound 38 in 120 mL of methylene chloride was added 270 mg (2.3 mmol) of methyl fluorosulfonate. The reaction mixture was stirred at room temperature for 3 hours. The precipitate was filtered off, washed with nethylene chloride (5 mL) to give 840 ng (99% yield) of quaternized pyridine 39 as white crystals.

NMR (DMSO-d 6) δ: 1.15 (3H, d, J = 5.8 Hz), 2.62 (3H, s), 3.2-4.4 (5H, m), 4.45 ( 3H, s), 4.60 and 4.12 (1H each, ABq, J = 9.2 Hz), 5.30 and 5.46 (1H each, ABq, J = 12.8 Hz) and 7.6 -8.9 (7H, m).

IC (KBr technology) gamma max: 3 400,: L 750 and 1590 -1 cm Analysis: for C ²⁴ H 24 ^N 3 ° 9 ^N 2 ^F calculated 49.14% C, 4.47% H, 7.13 % N, 11,43% WITH; found 49,56% C, 4.16% H, 7.26 % N, 11,03% WITH.

To a solution of Compound 39 (810 mg, 1.39 mmol) in tetrahydrofuran (100 mL) and ether (100 mL) was added pH 7.0 buffer (100 mL), followed by 10% palladium on carbon (750 mg). The mixture was hydrogenated ⁶⁰ m ^{even when} the nut 3 ¹ 0 psi on the Parr rubs ^P and ^C ce en ^d hundred m ^ ^s ^ tNOS (4 ^and 6 ° ^C).

The reaction mixture was filtered through celite and the catalyst was washed with ether (2 x 10 mL). The combined filtrate and wash solution was extracted with ether (2 x 40 mL) and lyophilized to give a yellow solid which was purified to give a yellow solid. BONDAPAK (Waters Associates) column (20 g), eluting with a yellow solid, which was purified on a BONDAPAK (Waters Associates) column (20 g), eluting with 5% acetonitrile in water at 55 kPa. Each 15 ml fraction was assayed by high pressure liquid chromatography and the fractions having ultraviolet absorption at lambda max 30 nm were combined and lyophilized to afford 141 mg (30% yield) of the title compound as a yellow solid.

NMR (D ₂ O) δ: 1.24 (3H, d, J = 7.0 Hz), 2.62 (3H, s), 3.2-3.5 (3H, m), 4.2- 4.4 (2H, m), 4.45 (3H, are, 4.50 and 4.59 (1H each, ABq, J = 12.16 Hz), 7.82 (1H, q, J = 7, 0 Hz, 6.5 Hz), 8.35 (1H, d, J = 7.0 Hz), and 8/65 (1H, d, J = 6.5 Hz).

^IR (KBr techn ^{i ka) g} max ama:

400, 1750 and ¹⁵⁸⁰ cm

UV λ max (H ₂ O): 296 nm (epsilon = 8

014).

Aialýza: for ^C 20 ^H j_7 ° 4 ^N 2 ^S 1 * ^1H 2 ° Calculated Found

% C, 57.85;% H, 5.85;

% C, 58.60;% H, 5.86;

7.94

7.87

N;

N.

Example

3- (2-Methyl-N-methylthiazol-4-ylmethanthio) -6alpha- [1- (R) -hydroxyethyl-7-oxo-1-azabicyclo (3.2.0) hept-en-2-ca

Preparation

^p -Nitrobenzyl-3- [2-methylthiazol-4-ylmethanthio] -6alpha- [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo (3.2.0) hept-2-ene-2-carboxylate

К ochozenému solution ⁽⁰ ° C) 1.4 ^g (4, ⁰ mrnob ketomezi ^p ro ^d u ^to the 5 in 12 ml aceeooitOlu was added 0.83 Mamo 4.5 ol) of diisopropylethylamine followed by 1.16 g (4, 3 mmol) of diphenylchlorophosphate in 2 ml of acetonitrile under nitrogen.

Vviúklý solution mícliá ³⁰ minutes ρι'ί 0 ° C and zdstá 7 ^^ 07 ^ 77 7--1116171: 027171oxy) aa- 6a ^and f 1- (R) - ^{h x} yddox YTH ^HL1 l ^{7 b} oxo 7ana tcykl.o ^{(third second 0)} teiJt- ene ^ ^ tarboxyUt. ^To this solution was added a solution of 0.83 ml (4.6 mmcjl) diisopropylethyl amine in 2 ml AccI-oo nirilu and then extended ^to the ^ ^{(0.62 g, 2} ^ ² rninm -eet ^the L-4-ec · saying t.oπm ^{p t} l ^{l-ltίtlιiano} u ^ ^p íipraiveněho ^p rocedure according to J. Amer. Chem. Soc. 71, 3570 (1949)] in 3 ml of ACE in her ril.

Reaction ^The organic solution is stirred ^{and 40} min. ^{Low: 14} ° C. ·· ^{S i!%} <N ^on the odfiJtruje and washed with ether (30 mL) to give 94 mg of 3 · 'in the title <· TOTM white solid.

NMR (CDC1 ₃₎ delta: 1.32 (3H, d, J = 7Hz), 2.68 (3H, s), 3.20 (2H, m), 3.76 (1H, d, J = 5 5 Hz), 4.16 (2H, S), 4.20 (1H, m), 5.40 (2H, q, J = 14Hz), 7.06 (1H, S), 7.68 ( 2H, d, J = 8Hz) and 8.24 (2H, d, J = 8Hz).

IR (KBr) gamma max: 3500, 1770 and 1700 cm

3- ^{(2-methyl-N-methyl thiazolidinone l-} 4 ^- ylmethyl hanth ^{t i o) -} 6a ^{LF -} [l ^{- (R) hydroxy} rox ^y et ^{hyj-oxo-L-azaMcyklo} (3.2 .0) hept-2-ene-2-carboxylate

To a solution of Compound 9 (525 mg, 1.1 mmol) in methylene chloride (20 mL) was added methyl fluorosulfonate (0.27 mL, 3.3 mmol). The reaction mixture was stirred at room temperature for 90 minutes. The precipitate was filtered off and washed with methylene chloride (50 mL) to give 650 mg (100% yield) of quaternizable thiazole 10 which was used for the next step without further purification.

Then, to a solution of compound 10 in 100 mL of tetrahydrofuran and 100 mL of ether was added 100 mL of pH 7.0 buffer, followed by the addition of 500 mg of 10% palladium on carbon. The mixture is hydrogenated at 20 psi for 45 minutes on a Paar shaker.

The reaction mixture was filtered through celite and the catalyst was washed with water (2 x 10 mL). The combined filtrate and washings were extracted with ether (2 x 100 mL) and lyophilized to give a yellow powder which was purified on a C ₁₈ BONDAPAK reverse phase column (8 g) (Waters Associates), combined with 5% acetonitrile in water at 55 ° C. kPa.

Each 15 nd fraction was assayed by high pressure liquid chromatography and ultraviolet n fraction. at lambda max 300 nm, they are combined and lyophilized. This afforded 145 mg (48% yield) of the title compound as a pale yellow powder.

NMR (CDCl 3): 1.20 (3H, d, J = 7Hz), 2.92 (3H, S), 3.08 (1H, d, J = 3.5Hz), 3.20 (1H, d) H, 3Hz), 3.44 (1H, dd, J = 1Hz, J = 3.5Hz), 4.00 (3H, 5), 4.20 (3H, m), 4.36 (2H) , m) and 7.88 (1H, s).

IR (KBr) gamma max: 3400, 1750 and ¹⁵⁸⁵ cm ^{@ 3} .

HV lambda max (Hy0); 296 nm (epsilon = 7,500).

Anal. Calcd for C 15 H 18 N 4 O 4 S 3 O 3 S Calculated 46.15 4 C, 5.64 ΐ H, 7.17 U N, 16.41% S;

found 4.50 t C, 5.26% H, 7.13% N, 16.20% S.

Example 9

Preparation of 3- (Ν, N'-dimethylimidazol-2-ylmethanthio) -6alpha- [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo (3.2.0) -hept-2-ene-2-carboxylate

CHo I ³

2) EtOH, A

HCl

HCl

2-mercaptomethyl-N-methylimidazole

To a solution of 10.4 g (58 mmol) of 2-chloromethyl-N-methylimidazole 31 [prepared according to J. Amer. Chem. Soc. 71, 383 (1949). In 200 ml of acetonitrile, 7.1 g (60 mmol) of N-acetylthiourea are added and the reaction mixture is heated under reflux for 90 minutes.

The precipitate was filtered off, washed with acetonitrile (20 ml) to give the isothiouronium salt, which was then dissolved in 120 ml of ethanol and refluxed for 18 hours under a nitrogen atmosphere. The reaction mixture was cooled to room temperature, concentrated in vacuo to a volume of about 60 mL, and the precipitate was filtered off.

Evaporation of the filtrate in vacuo gave 2-mercaptomethyl-N-methylimidazole 32 as a yellow oil, which was used for the next step without further purification.

NMR (D ₂ O) δ: 3.90 (3H, s), 4.10

OH

ČOjpNB

CH ₃

CO ₂ pNB

HCl p-nitrobenzyl-3- [N-methylimidazol-2-ylmethanthio] [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo (3.2.0) hept-2-ene-2-carboxylate

To a cooled solution (0 ° C) of 7.24 g (20.3 mmol) of ketone intermediate 5 in 35 ml of acetonitrile was added 2.8 g (21.3 mmol) of diisopropylethylamine in 2 ml of acetonitrile and then added under a nitrogen atmosphere of 5, 5 g (20.4 mmol) of diphenylchlorophosphate in 2 ml of acetonitrile.

The resulting solution was stirred at 0 ° C for 15 minutes and then a solution of 4.1 g (3.0 mmol) of diisopropylethylamine in 2 mL of acetonitrile was added followed by 4.6 g (31.0 mmol) of thiol 32. Reaction mixture was stirred at 0 ° C for 60 minutes.

The white precipitate was filtered off and washed with methylene chloride (20 ml) to give 6.6 g

(71% yield) the title compound as a white solid, m.p. 142 [deg.] C. NMR (DMSO-d 6) δ: 1.32 (3H, d, J = 7.0 Hz), 3.2-4.5 (5H, m), 3.2 (2H, s), 3.9 ( 3H s), 5.50 (2H, ABq, J = 14.0 Hz), 7.65 (2H, d, J = 6.5 Hz), 7.70 (2H, s) and 8.24 (2H, d, J = 6.6 Hz). IR (KBr technique) gamma max: 3,450, 1,770 and 1,690 cm Analysis: for ^c 21 ^h 20 ^N 4 ° 6 ^s 1 · ^{1 H} ₂ ° C, 52.18; H, 4.79; N, 11.59. found % C, 52.22;% H, 4.91;% N, 12.16.

t COjpNB

3- (N, N'-dimethylimidazol-2-ylmethanthio) -6alpha- [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo (3.2.0) hept-2-ene-2-carboxylate

To a suspension of Compound 33 (1.34 g, 3.0 mmol) in acetone (270 mL) was added methyl iodide (20 mL). The reaction mixture was stirred at room temperature for 4 days. The precipitate was filtered off, washed with acetone (20 ml) to give 1.70 g (96% yield) of quaternized imidazole 34 as yellow crystals, m.p. Mp 75-177 ° C.

NMR (DMSO-d 6)? 1.10 (3H, d, H = 6.2 (m / z), 3.30 (2H, s), 3.2-4.3 (6H, m), 3.95 (6H, s), 5.45 (2H, ABq, J 14 Hz), 7.65 (2H, d, J = 6.0) Hz). IR (KBr); gamma max: 3 00, 1750 and 16On cm ’.

Analysis for ^C ₂ ^1H 22 ^N 4 ° 6 1 ^S calculated 43.08% C, 9.60% H, 5.48% N;

found: C 43.02, H 9.02, N 5.44.

To a solution of 1.30 g (1.86 mmol) of compound 34 in 120 mL of tetarhydrofuran and 120 mL of ether was added 120 mL of pH 7.0 buffer followed by 900 mg of 30% palladium on celite. The mixture is hydrogenated for 40 minutes at 276 kPa on a Paar shaker.

The mixture was filtered through celite and the catalyst was washed with water (2 x 15 mL). The combined filtrate and washings were extracted with ether (3 x 100 mL) and lyophilized to give a yellow amorphous powder which was purified on a C 4 g BONDAPAK (Waters Associates) column (30 g) and eluted with 10% acetonitrile in water at 55 ° C. kPa.

Each 20 ml fraction was assayed by high pressure liquid chromatography, and the ultraviolet absorption fraction at lambda max 300 nm was combined by lyophilization to afford 220 mg (35% yield) of the title compound as a yellow powder.

NMR (d 6 O) δ: 1.12 (3H, d, J = 7.0 Hz), 3.08 (1H, dd, J = 13.0 Hz, 6.4 Hz), 3.15 (1H δ d, J = 13.0 Hz, 6.4 Hz), 3.45 (1H, dd, J = 3.2 Hz, 4.5 Hz), 3.85 (6H, s), 4.1- 4.3 (2H, m), 4.40 (1H, d, J = 13.5 Hz), 4.52 (1H, d, J = 13.5 Hz) and 7.40 (2H, s).

IR (KBr) gamma max: 3500, 1750 and 1590 cm

UV λ max (H 2 O): 296 nm (epsilon = 8,411).

Analysis for ^C | ig 5 ^H 3 ^N ° 4 ^S · H 2 O calculated 51.68% C, 5.67% H, 12.06% N, 9.50% S;

Found: C 49.93, H 5.94, N 11.46, S 9.03.

Example 10

P ^Rip rava ^3- (2, ^{3, 4-tri} methyl ^{th ylthio} azo ^l-5-yl met.han ^t h ^{i o)} 6 ^l alpha- [l- ^(R) H. ^yd roxy ^thy ^ - ^ oxo- ¹ -azabicyklof 3.2.0) hept-2-ene-2-carboxylate

WITH

1) '~ ^ · ΝΗ ₂

2) NaOH

2,4-dimethyl-5-mercaptomethylthiazole

To a solution of 4.8 g (26.0 mmol) of cilorslouron. ny 46: prepared according to the procedure of J. Amir.

Chem. Soc. 104.446 L (1 982) in 50 [mu] l of absolute thiourea were added 2.4 g (30 mmol) of thioamine. The reaction mixture is heated under reflux for 18 hours.

The precipitate was filtered, washed with ether (20 mL) to give isothicurcniová.sůl which extended ^p US ^children in 22 m ^{l IN} rosette h ^yd rox ^id u of ^{n e} and it is heated ^and at ¹⁰ 0 C 4 m ^{y and} nut in the atmosphere of nitrogen ^e Re.

The reaction mixture was then cooled to room temperature with 1N acid. pH was adjusted to 7.0 with hydrochloric acid and extracted with ether (3 x 50 mL). The combined ether phases were washed with water, brine and dried over magnesium sulfate. '

Evaporation of the dried solvent afforded 780 mg (49% yield) of thiol 47 as a colorless oil, which was used for the next step without further purification.

NMR (CDCl 3) δ: 2.05 (3H, s), 2.35 (3H, s)

HE

δθ £ ρΝΒ

OH

COjpNB ^{p CH} 3 -N ^{IT-nitrobenzyl} 3 - [2, ^{4-D i} meth ^yl AZC th ^{i l} - ^5-yl -methanthio] ^lf 6a and Q ^l - (R) -h ^{y y} Hydroxy ethyl] - 7-oxo-bicyclo (3.2.0) ^{hept-2-ene-2-carboxylate} alpha ^hl azenému solution (0 ° C) ^{1.4 g (4-mmol) and the} etomez ^to the producer 5 ² 5 610 ml (4.7 mmol) of diisopropylethylamine in 1 ml of acetonitrile was added to 1.1 ml of acetonite, and 1.15 g (4.3 mmol) of diphenylchlorophosphate in 1 ml of acetonitrile was added under a nitrogen atmosphere.

The resulting solution ^was stirred ^for 20 minutes ^at 0 ° C and ^{added to} 61.0 m ^g (4.7 mmol) of diisopropylethylamine in 1 ml of acetonitrile and finally 750 mg (4.7 mmol) of thiol 47 was added. in ² mL of ACE they ^{t t} r u ^{il. CZK R} e n ^e was stirred for 3 he ^di n ^{y at} 0 ° C.

The precipitate was filtered off, washed with methylene chloride (20 mL) to give 1.14 g (61% yield) of the title compound as a white solid.

NMR (DMSO-d 6) δ: 1.25 (3H, d, J = 6.4 Hz), 2.30 (3H, s), 2.65 (3H, s), 3.1-3.4 ( 3H, m), 4.10 (1H, broad s), 4.0-4.5 (3H, m), 5.25 and 5.50 (1H each ABq, J = 4 Hz), 7.68 ( 2H, d, J = 8.5 Hz) and 8.25 (2H, d, J = 8.5 Hz).

IR (KBr) ymax: 3500, 1770 and 1690 cm ^1st

Analysis: for ^C 22 ^H 23 ^N 6 ^S 3 ° 2 requires 53.73% C, 4.71% H, 8.57% N, 13.44% S;

found: C 53.97, H 4.74, N 8.58, S 13.10.

3 ^{- (2,} 3, ⁴ -trimet ^HYL iazo t ^{h l "of 5} - ^y o ^h-met on ant ^hi) - ⁶ and ^LF N- [l- ^(R) - ^{hydroxyacetophenone Y} et ^hyl] -7- J. ^oxo-aza-b ic ^y bicyclo (3.2.0) hept-2-ene-2-carboxylate

To a solution of Compound 48 (1.97 g, 4.0 mmol) in methylene chloride (180 mL) was added a solution of methyl fluorosulfonate (0.98 mL, 13 mmol) in methylene chloride (2 mL). The reaction mixture was stirred at room temperature for 70 minutes.

The reaction mixture was poured into a solution of ether (400 mL) and n-pentane (100 mL). The precipitate was filtered off and washed with ether (20 ml) to give 1.6 g (65.5% yield) of quaternized thiazole 49 as a white amorphous powder.

NMR (DMSO-d6) δ: 1.25 (3H, s, J = 6.5 Hz), 2.45 (3H, s), 2.80 (3H, s), 3.2-4.5 ( 6H, m), 3.90 (3H, s), 5.30 (2H, broad), 7.60 and 8.2 (1H each, d, J = 8.5 Hz).

IC (KBr technique) gamma max: 3400, 1770 and 1690 cm

Analysis: · ^1/2 H2 ⁰ Calculated 45.09% C, 4.44% H, 6.86% N;

Found: C, 44.50; H, 4.38; N, 6.58.

To a solution of 1.0 g (1.72 mmol) of compound 49 in 100 mL of tetrahydrofuran and 100 mL of ether was added 100 mL of pH 7.0 buffer, followed by 1.0 g of 10% palladium on carbon. The reaction mixture is hydrogenated for 40 minutes at 276 kPa on a Paar shaker.

The mixture was filtered through celite and the catalyst was washed with water (2 x 10 mL). The combined filtrate and washings were extracted with ether (3.100 mL) and lyophilized to give a yellow powder which was purified on a Cug BONDAPAK (Waters Associates) column (40 g), eluting with 10% acetonitrile in water at 55 kPa.

Each 15 ml fraction was assayed by high pressure liquid chromatography and the ultraviolet absorption fractions at lambda max 30 nm were combined and lyophilized to afford 315 mg (50% yield) of the title compound as a yellow solid.

NMR (D ₂ O) δ: 1.25 (3H, d, = 7.0 Hz), 2.25 (3H, s), 2.90 (3H, s), 3.0-3.30 (3H) , m), 3.90 (3H, s) and 4.1-4.4 (4H, m).

IR (KBr) ymax: ^{3 4} 00, ¹⁷⁵⁰ and ^{1580 cm} '\ UV .lambda.max: 0 1 _2: 297 nm (epsilon = 8994).

H, 6.09; N, 7.79. Found: C, 47.96; H, 5.83; N, 7.89.

Example 11

Preparation of 3- [2- (N-methylthiazolium) methylthio] -6alpha- [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo ((3.2.0) hept-2-ene-2-carboxylate)

soci ₂

-

2-mercaptomethylthiazole

To a chloroform solution (30 mL) thionyl chloride (3.81 mL, 0.052 M) was added at room temperature ^{3.60 g (0, 02} -6 ^M) hydrox ^y me ^th yl ^th thiazol on EZ ^{2 No h} o n ^{y di h} for iv ^and at ⁵⁰ ° C.

Evaporation of the chloroform in vacuo gave a brown solid which was dissolved in 30 ml of absolute ethanol.

Then 2.04 g (0.026) of thiourea ^y. The mixture was then heated to reflux for 18 hours. The precipitate was filtered off, washed with ethanol and ether to give 3.4 g (55% yield) of the isothiouronium salt 3.

The isothiouronium salt 2 ^is dissolved in 30 ml of water and purged with nitrogen for 20 minutes. Then ^a hydride ^{and 1.10 g (0 027} M), h ^yd Roxi ^d u n ^{d s} with him, and the mixture was ^h iv ^and 2 m ^{and y} nut to 100 ° C. ^The organic solution was cooled to 0 ° C when ^y is ^l acetic another at Ravi ^{P P} H to ^ ^0, then ^p Roveda extratae ethyl acetate (2 x 35 mL).

The organic layer was dried over magnesium sulfate and evaporated in vacuo to give 0.75 g (42% yield) of thiol 4 as a yellow oil which was used without further purification.

NMR (CDC1 ₃₎ delta: 2.1 (1H, t), 4.0 (2H, d, J = 10Hz), 7.27 (1H, d, J = 2.0 Hz) and 8.85 ( 1 H, d, J = 2.0 Hz).

OH

COjpNB

OH

CO ₂ pNB-nitrotenzyl-4-L-thiazol-4-yl-methyl-4 - [(R) -hydroxyethyl] -1H-7-oxo-1-azabicyclo (3.2.0) hept-2-ene-2-carboxylate ^the ocHazen roztolku ^ nu ⁰ O ^1.4 g (4 mmol ^! ^{e t p} genus ^kt be confined in ^{5 l} acetone ⁸ m ^ rhu 0.79 ml (4.4 mmol) of diisopropylethylamine followed by the addition 1.17 g (4.4 mmol) of diphenylchlorophosphate under a nitrogen atmosphere.

^The Zn ^IKL characterized ^extensible by ^k m ^s chá 30 minutes: ^{eg 0} ° ^C to ignite in ^the JD-n-rolDenzyM-fd ^ en ^ ^ osforidox -6- [l- (R) -hydroxyethyl] -7 oxo-1-azabicyclo (3.2.0) hept-2-ene-2-carboxulate. To this solution was added a solution of 0.79 mL (4.4 mmol) of diisopropylethylamine in 2 mL of acetonitrile, followed by a solution of 0.72 g of thiol 4 in 2 mL of acetonitrile.

Rea ^{at the} CN ^s roztoJc with m ^I CH ^{and 60m and} nu ^{t at 0} ° ^C and ^p Otom of a ^row of ^{copper 50} m ¹ e ^ hceUU and ^p rom ^y is 30 mL water, 20 mL of 10% aqueous phosphoric acid and 30 ml of sodium chloride solution. Evaporation of dried (MgSO3) solvent gave a crystalline solid which was triturated with ether and ^the ZIS Å ⁽⁷⁸² mg, 42% yield ^of the ^) produHu ⁶ in the form ^{of WHT hk} YC r ^y s ^{t L} and vi, ^{i. t} . , ^158-160 ° ^O.

NMR (CDC1 ₃₎ delta: 1.32 (3H, d, J = 7.0 Hz), 3.28 (3H, m), 4.20 (2H, m), 4.36 (2H, s) 5.40 (2H, d), 7.40 (1H, d, J = 4.0Hz), 7.64 (2H, d, J = 8Hz), 7.76 (1H, d, J = 4), 0 Hz) and 8.24 (2H, d, J = 8 Hz).

IR (KBr technique) gamma max:

500, 1,770 and 1,700 cm Analysis: calculated for C ₂₀ H ₁₉ M ₃ 3 ₆ 6 ₂ 52.05% found 52.35%

O, 4.15%

H, 9.10% N,

H, 8.72% N,

O, 4.40%

About me

CO ₂ pMB

13,89%

13,90%

WITH;

WITH.

Pd / C, H_

FSO3

IN

3- (2- (N-methylthiazolium) methylthio) -6α? - [1- (R) -hydroxyethyl] -7-oxo-O-azabicyclo (3.2.0) hept-2-ene-2-carboxylate To a solution of 782 mg (1.36 mmol) of compound 6 in 55 ml of methylene chloride was added 0.5 methylfluorophonate and stirred at room temperature for 90 minutes, and the precipitate was filtered off, washed with methylene chloride (30 ml) and ether (20 ml) to give 630 mg of crude quaternized thiazole 7, which was used for the next step without further purification.

To a solution of compound 7 in 140 µl tetrahydrofuran and 120 µl ether was added 140 ml of pH 7.0 buffer followed by 650 mg of 10% palladium on carbon. The reaction mixture was hydrogenated at 20 psi on a Paar shaker for 35 minutes.

The mixture was filtered and the catalyst was washed with water (2 x 10 mL). The combined filtrate and washings were extracted with ether (2 x 150 mL) and lyophilized to give a yellow powder. The crude yellow powder was purified on a BONDAPAK reverse phase column (7 g) (Waters As ™), eluting with 5% acetonitrile in water at 55 kPa.

Each 15 ul fraction was assayed by high pressure liquid chromatography, fractions having an ultraviolet absorption at Amax 300 nm were collected and 0ycfi0izací _^ afforded 23 mg (5% yield) of the title compound as a yellow amorphous solid. NMR (d 6 O) δ 1.28 (3H, d, J = 7.0 Hz), 3.12 (2H, d, J = 7.0 Hz), 3.44 (1H, dd,

J = 1.0 Hz and 3.0 Hz), 4.20 (3H, s), 4.24 (2H, n), 4.76 (3H, n), 8.12 (1H, d, J = 4 Hz) and 8.24 (1H, d, J = Hz).

IR (KBr) gana max: 3,400, 1,740 and 1,580 cm ^-1 .

UV λ max (H ₂ O): 292 nm = 7.885).

Example 12

™ 3

S-CH-θ-CH

P ^Rip rava ^{3- (1-} (RS) methyl ^^^ - ^ p ^ neth ridi n ^^^ ^{ehantliio] -6} a] .I ^ C ^ -L ^l - (R) -h ^yd RCX ^s e ^th У ^o ] -7

^{-cxo-1-aza-yl} bicyclo (3.2.0) hept-2-ene-2-carboxylate

OH

₂ pNB

CO ₂ pNB p-nitrobenzyl-3- [] - (R, S) methylpyridin-3-ylmethanthio ^] -6alpha- [1- (R) hydroxyethyl] -7-oxo-1-azabicyclo (3.2.0) hept- 2-tn-2-carboxylate

The oc and ^y ^ y ^ t ^ i ^ CMU solution (0 ° C) ^{(1.85 g,} 5 ³ mmol) om ^{t to} t ^ t ^ ^{y. P} ro ^d u u 5 t ^{kt 2} 0 ml асеЛопйгИи We added 754 mg (5.8 imnol) in 1 mL diisopropylehhylaminu acetoniirilu, before a solution - 57 g (5.84 mniml) of diphenyl chlorophosphate in 2 ml of acetoníirilu nitrogen atmooféře .

Vzynklý mu ^ solution and ¹⁵ min ^at 0 ° C and [then ^{p Class The} organic solution ⁽⁷⁵⁴ mg MMC ^5.8> l) in 1 ml dřisopropylethylaminu acetoniirilu and then 814 mg (5.8 mm) of) thiol 27 in 2 ml of acetonitrile. After my ^ and ^{3 hr} Odin ^yp R.I ⁰ ° C and ^ are the SMAS reatórní et diluted with ²⁰⁰ ml. ^hy Lacet ^at u, washed with ice-cold brine (200 mL), water (200 miles aqueous sodium hydro genuuhičitanu * (-00 mL) and brine (100 mL).

Evaporation of the dried (MgSO4) solvent afforded a yellow oil which was purified by silica gel column chromatography, eluting with 50% acetone and 50% loride to give 1.65 g of the title compound as a yellow solid.

NMR (CDCl3) δ: 1.22 and 1.25 (3H each d, J = 7.0 Hz),

J = 7.2 Hz, 2.4-3.3 (3H, m, 3.8-4.2 (3H, m), 5.35 (2H, ABg,

1.46 and 1.50 (3H each d,

J = 14.5 Hz) and 7.2-8.6 (8H, m.

IR (KBr) λ max: 3,400, 1/65 and 1,690 cm ³ .

Analysis: Calculated for C ₂ ^H 3 ^N 3 ^O 3 ^S 58.83 Found 57.15% C, 4.94% C, 5.04

OH

H,

H,%

2) NaOH

N, 6.83

N, 8.28

SOCl -% S;

% S.

HCl

4- (1'-Cercaptoethyl) pyridine

To a solution of 25 g 1 - ^(4-p-y ridyIh thanohi 25 ^ připaaen.nécu procedure of J. Chem. Suc., Perkin II, 1462 (1974)] in 100 ml of chloroform was added 50 g thinnylchlnridu. The mixture was The mixture was heated at reflux for 2 hours.

Evaporation of the solvents in vacuo gave chloro compound 26 as a semi-solid which was used for the next step without further purification. To a solution of 26 in 160 mL of ethanol was added a hot solution of 14.4 g of thiococoaine in 75 mL of ethanol.

The reaction mixture was heated to reflux for 18 hours. The ethanol was evaporated and the remaining stream was dissolved in 100 ml of water and the pH was adjusted to ID by addition of 2N NaOH. The mixture was stirred at room temperature for 90 minutes, adjusted to pH 6.0 with 6N HCl and extracted with ether (x2). 200 ml).

Evaporation of the dried (MgSO 4) solvent afforded a yellow oil which was distilled at 666 Pa and boiling at 60-65 ° C to give 11.0 g (38% yield) of pure thiol 27 as a colorless oil.

NMR (CDCl ₃ ) δ: 1.70 (3H,

6.0 Hz, 5.8 Hz), 7.20 (2H, d, d, J = 6.0 Hz), 2.05 (1H,

J = 6.2 Hz) and 8.5 (2H, d, d, J = 5.8 Hz)

J = 6.2 Hz).

4.20 (1 H, t,

3- [1- (RS) -methyl-N-methyl-pyridin-3-yl-methanthio] -6alpha- [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo (3.2.0) -hept-2 -en-2-carboxylate

To a solution of compound 28 (1.1 g, 2.34 mmol) in acetone (100 mL) was added methyl iodide (10 mL). The reaction mixture was stirred at room temperature for 18 hours. The precipitate was filtered off, washed with methylene chloride (10 ml) to give 1.4 g (100% yield) of quaternized pyridine 29 as a yellow powder.

NMR (DMSO-d 6) δ: 1.10 (3H, d, J = 6.5 Hz), 1.62 (3H, d, J = 7.5 Hz), 2.6-4.2 (6H, 1I), 4.39 (3H, s), 5.42 (2H, ABq, J = 13.6 Hz) and 7.9-9.2 (8H, m).

IR (KBr) λ max: 3400, 1770 and 1190 cm

Analysis calculated for: C ₂ = H ₂₆ N ₃ O ₍₁ S 1 ₃ calculated 47.14% C, 4.29% H, 6.87% N, 5.24% S;

found 47.19 C, 4.78% 11, 6.11% N, 5.41 4 S.

To a solution of 1.45 g (2.37 mmol) of 29 in 120 mL of tetrahydrofuran and 120 mL of ether was added 120 mL of pH 7.0 buffer followed by 1.5 g of 10% palladium on carbon. The reaction mixture was hydrogenated at 310 kPa (60 psi) for 60 minutes on a Paar shaker. The mixture was filtered through celite and the catalyst was washed with water (2 x 15 mL).

The combined filtrate and washings were extracted with ether (2 x 200 mL) and lyophilized to give a yellow solid which was purified on a reverse phase BONDAPAK (Waters Associates) column (50 g), eluted with 5-acetonitrile in water at 55 kPa .

Each 20 ml fraction was assayed by high pressure liquid chromatography and the ultraviolet absorption fractions at lambda max 300 nm were combined and lyophilized to afford 200 mg (24% yield) of the title compound as a yellow amorphous solid.

NMR (D ₂ O) δ 1.32 (3H, d, J = 7.0 Hz), 1.63 (3H, d, J = 7.2 Hz), 2.5-4.6 (6H, m),

4.32 (3H, s) and 8.2-8.9 (4H, m).

IR (KBr)) ymax: 3400, 1750 and 1590 cm ^3rd

UV λ max (H ₂ O): 296 nm (epsilon = 7573).

Analysis: for ^C ₁₇ ^H 20 ^N 4 ^S 2 l ° ¹ ° 2 ^{1.2 H} Calculated 54.38% C, 5.77% H, 7.46% N;

found

Example

Preparation of 3- (N-methyl-N'-benzylimidazol-2-ylmethanthio) -6alpha- [1- (R) -hydroxyethyl-7-oxo-1-azabicyclo (3.2.0) hept-2-en-2- carboxylate

1)

WITH

H, N-Ac

N-benzyl-2-mercaptomethylimidazole

To a solution of 3.23 g (13.0 mmol) of chloro compound 41, prepared according to the method of J. Amer.

Chem. Soc. 71, ³⁸³ (1949)] in 80 mL of acetonitrile was added 1.72 g (14.5 mmol) of N-acetylthiourea. The reaction mixture was refluxed for 3 hours.

The precipitate was filtered off and washed with acetonitrile (10 ml) to give the isothiouronium salt, which was then dissolved in 80 ml of absolute ethanol and refluxed for 18 hours under a nitrogen atmosphere.

The reaction mixture was cooled to room temperature, concentrated in vacuo to a volume of about 30 mL, and the precipitate was filtered off. Evaporation of the filtrate in vacuo gave 3.5 g (97% yield) of thiol 42 as a yellow thick syrup.

NMR (CDC1 ₃₎ delta: 2.1 (III t .1 - 4, ^r Hz), 3.80 (211, s), 5.20 (2 (1H, s) and 6.8-7. Δ (7H, m).

ОН

(AND

C1-P (O0) ₂

CO ₂ pNB

^ n ^ r ^ ^^ enzymes Enz fN ^ ^^^ Azoia .- ^ - ydrnethhnthiiJ-ťjalfa- ^-] .- (R) ^{hydroxypropoxy} ° X ^Y et ^hyl J ^-7- oxo ^-l- azabicyclo ⁱ cyclo (3.2 .0) hept-2-ene-2-carboxylic ° x ^y acrylate ^{to a} cooled solution ⁽⁰ ° C) @ ^{3.03 g, 5} mmol). tetomez ^ roduHu п ⁵ in ^{70 1} of acetonitrile was added 1.17 g (9 Diisopropylethylamine in 2 ml of acetonitrile, then 2.4 g (9.0 mmol) of diphenylchlorophosphate in 2 ml of acetonitrile are added under a nitrogen atmosphere.

MMN ⁱ k ^l ý solution my ^ HA ^{3 20} Mbuti ^{at 0} ° C and ^p Otom the ^{p Class} solution 1Д ⁷ g (9, ⁰ mmol ¹⁾ DIIS ° prop ^{ylethynyl y} Lanina in 2 ml of acetonitrile, whereupon 4.8 g (15 mmol) of the thiol 42. Thereafter, a further 1.93 g (15 mmol) of diisopropylethylamine and the mixture was stirred for 2 h ^{and y} n ^at 0 ° C.

The precipitate was filtered off and washed with cold methylene chloride (20 ml) to give

2.5 g (55% yield) of the title compound as a white solid.

NMR (DMSO-d 6) δ: 1.23 (3H, d, J = 7.2 Hz), 2.5-4.1 (6H, m), 4.25 (2H, s), 5.20 ( 2H, s), 5.20 and 5.45 (1H each d, J = 14.5 Hz) and 6.9-8.3 (11H, m).

IR (KBr) ymax: 3400, 1775 and 1690 cm ^1st

3- ^J N-meth ^yl -N' ^{Yli-benz imidazol-2-yl l-} nethanth ⁱ °) ^- 6a ^LF a- ^fl-J R) ^- h ^y dr ° xyothy ^{l] -7-ox} ° -l azabzlcyúclo (3.2.0) hept-2-ene-2-carboxylate

To a solution of Compound 43 (1.76 g, 3.3 mol) in methylene chloride (1.1 L) was added 1.15 mL of the mixture and stirred at room temperature for 2 hours.

about 15 ml. The precipitate was filtered off, washed with g (74% yield) of quaternized imidazole 44 (13.4 mmol) methyl fluorosulfonate. The reaction mixture was concentrated in vacuo to a volume of methylene chloride (10 mL) to give 1.58 as a white solid.

NMR

ABq, J = (DMSO-d6) [delta]: 1.15 (3H, d

10.8 Hz), 5.24 and 5.46 (1H each, ABq, J = 14

J = 7.0 Hz), 3.2-4.4 (6H, m), 4.70 and 5.0 (1H each,

Hz), 5.50 (2H, s) and 7.4-8.4 (11H, m).

^iC (KBr • technology) gamma max:

500 ^, 1,770 ^and 1,700

- 1 cm

Analysis for 8H2 O _{2 g of} N ₂ F ₄ O9S Calculated Found

51.48% C, 4.47%

51.84% C, 4.52%

H, 8.67% N, 10.20%

H, 8.65% N, 9.87%

WITH;

WITH.

To the solution was added 120 45 minutes at room temperature

1.11 g (1.71 mmol) of compound 44 in 100 mL of tetrahydrofuran and 100 mL of ether, with a pH 7.0 pH buffer, 310 kPa and 1.0 g of 10% palladium on carbon was added. The mixture was hydrogenated on a Paar shaker.

Celite and the filtrate and wash water were extracted with acetonitrile powder

The mixture was filtered through a catalyst and washed with water (2 x 10 mL). Combined with ether (2 x 70 mL) and lyophilized to give a yellow purified on a Cm BONDAPAK (Waters Associates) column (40 g), eluting with 10% water at 55 kPa.

the fraction was assayed by high pressure liquid chromatography and the ultraviolet absorption fraction at lambda max 30 nm was combined and lyophilized to afford 305 mg (43% yield) of the title compound as a pale yellow amorphous solid.

Each 15 ml

NMR (DMSO) δ: 1.40 (3H, d, J = (2H, m), 4.23 (2H, broad s), 5.57 (2H,

7.0 Hz), 2.9-3.4 (3H, m), 3.98 (3H, s), 4.0-4.2 s) and 7.2-7.65 (7H, m) .

IR (KBr technique) gamma max:

UV λ max (H ₂ O): 299 nm

400, (epsilon ~ 8

760 and 1590 cm 807).

Analysis: for C ₂ H 23 N ₃ O ₄ S 1

1/2 HnO calculated

57.25% C, 5.94

C 56.66, 5.70

H, 9.54

9.49

H,

N,

7.28

8.30

WITH;

WITH.

Example

Preparation

3- (-methyl-N-methylpyridin-1-ylmethananthino) -4-alpha-1- (R) -hydroxyethyl-7-oxonazabicyclo (3.2.0) -hept-2-ene-2-carboxylate

LÍA1H

HCl

2-methyl-3-mercaptomethylpyridine

The ester 12 was prepared according to the procedure of J. Org. Chem., 21,800 (1956). To a cooled suspension had roztyku ^{d (0} ° ^{C) 2.86} g H ^ u ^ ummumhydňclu in ⁵⁰ m ¹ of anhyd ^{eh t} e ^t rah ^yd ro ^furan was added dropwise a solution of 6.23 g (0.038 M) of the ester 12 in 15 ml of tetrahydrofuran over 15 minutes. After ^I C m ^{hook 60} m nu ^{i t at 0} ° ^C and ^p Otom the straight ^and d ^{1 and 50m} et ^hyl acetate, ^and here.

The precipitate was filtered off and washed with a saturated aqueous ammonium chloride solution. The organic layer was dried over magnesium sulfate, filtered, and evaporated in vacuo to give 3.2 g (70% yield) of hydroxymethylpyridine 13 as a yellow oil.

NMR (CDCl 3) of δ delta: 2.46 (3H, S), 4.73 (2H, S), 5.1 (1H, broad, 7.2 (1H, dd, J = 8 Hz), 7, 8 (1H, dd, J = 8Hz, J = 1Hz) and 8.3 (1H, dd, J = 7Hz, J = 1Hz) and 8.3 (1H, dd, J = 7Hz, J) = 1 Hz). · * ^K oc ^hl Azen ^s him roztob ⁽⁰ ° ^C) of 4 m ^{1 thi} ones ^l ch ^l or ^even du v · 10 m ^{1 methylcarbazole} ench ^l or ^id u ^{EXAMPLE IK P} e A solution of 3.2 g (0.026 M) of alcohol 13 in 10 ml of methylene chloride under a nitrogen atmosphere for 15 minutes, the cooling bath was removed and the reaction mixture was left at room temperature for 3 hours.

All solvents were evaporated in vacuo to give compound 14 as a brown solid, which was used for the next step without further purification. The crude brown solid was dissolved in 30 mL of absolute ethanol.

^P a ^t OM was ^{added 2.5} g of ^{(D, M 32 °) No} omo ^Thi s ⁱ n ^y and sires with sealing ^18h of ^di n at ^65-70 ° ^C. The mixture was cooled to room temperature. The precipitate was filtered off and washed with ethanol (20 ml) and ether (50 ml) to give 30 g of the isothiouronium salt.

This salt was dissolved in 10 ml of water under a nitrogen atmosphere was added a solution of 640 mg (0.016 M) ^{hydroxyacetophenone id} for sodium ^eh s 10 m ^{1 y} waters. ^R EAK ^{N NU} mixture was Hv ^{h and 2M and the} nu 1 ⁰ ° C and then octáadí to 0 ° C and acetic acid adjusted to pH 6.0 and extracted with chloroform (2 mL x.35 ). Evaporation of the dried (MgSO4) chloroform gave 941 mg (46% yield) of thiol 15 as a yellow oil. NMR (ν M Dc CI, ______________ _ _r , _r (1H, dd, JR = 8 Hz), 7.57 (1H, dd, J = 8 Hz)

NMR (CDCC, δ thiol 15 delta): 1.8 (1H, t), 2.60 '(3H, S), 3.73 (2H, d, J = 10 Hz), 7.13 and 8.43 ( 1H, dd, J = 8Hz, 3Hz).

2)

OH

1S ^{p- and} n Trob ^{(NZ-yl-3- (2 -} ime ^{phenyllithium} R ^d in ^{- 3-yl} methyl ant ^{th hi} o) ^{- f} and 6Al ^- [1- (R) ^{-hydroxy-hydroxy-eth y y} Lj -7-oxo-bicyclo (3.2.0) ^{hept-2-ene-2-carboxylate} oc ^h bath ^E mites ^to him at (0 ° C) ^{1.52 g (4, 3} 7 mmol) to yield ^ ^ u ^ genus 5 in 5 ml of acetonitrile ^and Lu are added 0.86 ml (4.80 mmol) of diisopropylethylamine, and then a solution of 1.17 g (4.37 mmol) of diphenyl chlorophosphate in 3 mL of acetonitrile under nitrogen atmosphere.

Ignites ^{the extensible} lysed by ^k m c ^{s hook 30} m nu ^{i t at 0} ° ^C to ^I of the PN ^{KA IT} ro ^b enz ^yl-3- (d ^{if y} en LFOs ^f oryl ^This mL of acetonitrile ox ^y ) ^- 6a ^-f1 - (R) -h ^yd roxy ^{th yl] -7-oxo-l -} azabicyclo (2.3 ^{0) h} ept-2-ene-2 ^to arboxylát. a solution of 0.86 ml (4.80 mmol) of diisopropylethylamine in 2 and then 940 mg (6.76 mmol) of thiol 15 in 2 ml of acetonitrile is added.

^R ea ^{CZK tup} was mí.ch ^{and 60m and} nu ^{t at 0} ° C. ^RA precipitate is ^F (30 mL) to give 1.12 g (55% yield) of solid la ^{t ky, mp. 186} DEG- ¹⁸⁸ DEG ^{C. (} dec. ⁾ .

It filters ^{out the} title in a ^{p y y} is etlierein form light yellow

NMR (DMSO-d 6) δ: 1.20 (3H, d, J = 7 Hz, 2.60 (3H,

4.32 (2H, s), 5.16 (1H, d, J = 5 Hz), 5.44 (2H, d)

J = 8 Hz), 8.36 (2H, d, J = 8 Hz) and 8.48 (1H, <

[, q, J dd, J =

S), 3.40 (m, 2H), 4.16 (m, 2H) 14 Hz), 7.32 (2H, m), 7.8 (2H, d, 5.5 Hz, 1.5 Hz) ).

IR (KBr technique) gamma max:

500, 1 770 and 1 750 ^cm ' ¹

Analysis: for ^C 23 ^H 24 ^N 3 ° 6 ^N

4.94%

4.99% calculated

СО ₂ рМв /

3- (2-methyl-N-methylpyridine (3.2.0) hept-2-ene-2-carboxylate)

To a solution of ^{697 g (1.19} mmol) of compound ^{16 was} added dropwise 0.5 ml (6.18 mmol) of methyl fluorosulfonate at room temperature. The precipitate was filtered off, 777 mg (90% yield) of quaternized pyridine in 100 ml of methylene chloride was stirred at 10 ° C for 10 minutes. The mixture was stirred for 2.5 hours, washed with 30 mL of methylene chloride to give 17 as a yellow solid.

NMR (CDCl ₃ ) 17 δ: 1.20 (3H,

4.16 (2H, m), 4.60 (2H, s), 5.20 (1H, m), 5.42 (2H, q, J d, J)

Hz), 2.82 (3H,

Hz), 7.80

s), 4.36 (3H, s), (2H, d, J = 8Hz),

8.04 (1H, dd, J = 7Hz, 6.5Hz), 8.32 (2H, d, J = 8 Hz), 8.64 (1H, d, J = 7.5 Hz) and 9.08 (LH, d, J = 7.5Hz). IR (KBr technique) gamma max: 3 500 and 1 765 , 1 cm. Analysis: for C ^ HH ^NN -O ^S calculated: 48.91% C, 4.55% H, 7.23% N, 11.04 % S; found C 49.39, 3.97% H, 7.20% N, 10.98 % S.

To solution 1.10. g (1.88 mmol) of compound 17 in 80 ml of tetrahydrofuran and 80 ml of ether was added 80 ml of pH 7.0 buffer followed by 800 mg of 10% palladium on carbon. The mixture is hydrogenated at 20 psi on a Paar shaker for 40 minutes.

The mixture was filtered through celite and the catalyst was washed with water (2 x 10 mL). The combined filtrate and washings were extracted with ether (2 x 100 mL) and lyophilized to give a yellow powder, which was purified by HP-20 column chromatography, eluting with water and 5% acetonitrile in water.

Each 15 ml fraction was assayed by high pressure liquid chromatography and the ultraviolet absorption fractions at lambda max 30 nm were combined and lyophilized to give 614 mg (42% yield) of the title product as a pale yellow powder.

NMR (D ₂ O) δ: 1.28 (d, 3H, J = 7Hz), 2.86 (3H, s), 3.20 (2H, dd, J = 10Hz, 3.5Hz), 3.42 (1H, dd, J = 5.4 Hz, 3.5 Hz), 4.20 (3H, m), 4.32 (3H, s), 4.35 (2H, S), 9 88 (1H, dd, J = 7.2 Hz, 6.5 Hz), 8.5 (1H, d, J = 8 Hz) and 8.70 (1H, d, J = 8 Hz). -1

IR (KBr) ymax: 3400, ^{1760 1 5} 9 ⁰ cm.

UV λ max (H ₂ O): 298 nm (epsilon = 8,391).

For C ₁₇ H ₂₀ F ₂ O ₃ S.H ₂ O calculated C 55.73, H 5.46, N 7.65, S 8.74;

Found: C 55.50, H 6.05, N 7.74, S 8.68.

Example 15

Preparation

3- [4- (N, N-dimethyl-1,2'-triazolinyl) methylthio] -4-α- (R) -hydroxyethyl] -7-oxo-1-azabicyclo (3.2.0) hept-2-en-2 -carboxylate

To the ice

d) H 2 / Pd-C cooled stirred solution of 4- (methanthioacetate) -1-methyl-1,2,3-triazole (590 mg, 3.52 mmol) in anhydrous methylene chloride (2 mL) was added dropwise under nitrogen atmosphere methyltrifluoromethanesulfonate ( 0.1 ml (5.16 mmol).

After half an hour the bath was removed and after 1 hour the solvent was removed by suction. The residual oil was dissolved in a few ml of water and the solution was cooled in an ice bath. A cold solution of sodium hydroxide (305 mg, 7.59 in a few ml of water) was then added and the reaction mixture was stirred for 3/4 hours.

The solution is diluted with 25 ml of water and the pH is adjusted to 7.5 by addition of solid monosodium phosphate. Then 14 ml of this solution (about 1.9 mmol of triazol-4-ol) are added to an ice-cold enol phosphate solution (1.0 g, 1.0 ml). 72 mmol). in tetrahydrofuran (10 mL).

The reaction mixture was stirred for 3/4 hours (during this reaction a certain crystalline substance, probably Na 2 HT 3 O) precipitated). The slurry was placed in a pressure vessel between tetrahydrofuran (20 mL) and water (20 mL). Ether (30 mL) and 10% palladium on carbon (1.0 g) were added and the mixture was hydrogenated at 50 psi for 1 hour. The organic phase was separated and washed with water (2 x 5 mL).

The combined aqueous phases were filtered and the filtrate was bent under high vacuum (1 mm Hg, 1.5 h). The yellow solution is then chromatographed (medium-thickness reverse phase column, 35 x 35 x 90 mm, water as eluent) to give, after lyophilization, 395 mg of cryptene slightly contaminated with some inorganic material.

The material was purified by high pressure liquid chromatography (10 x 300 mm Waters Microbondapack C18 column, multiple dosing, water as eluent) to give 310 mg (57% yield) of isomers A as a tan powder.

^L-NMR (D ₂ O) delta: 1.23 (3H ^d, J = ^{6, 4Hz), 3,} 10 (2H, d, J = ^9, 1 ^{Hz), 3} 24 (1H, ^q,

J = 2.7, 6.1 Hzb 4.03-4.71 (10H, o), 8.46 (1H, s).

IR

760 c ^m

UV (phosphate buffer pH 7.4, M = 0.05) .lambda.max 296 ([.alpha.] D @ 20 = 7.500).

B. Preparation of B 1 -Me isomers (likely structure) and C isomers

a) MeOTf

b) aqueous NaOH

OH

CO ₂ pNB

d) _H2 / Pd-C

To an ice-cooled solution of 4- (m-holpholyl) -2-m-thiol, 2,3-tritzil (1.20 g,

7.02 mmol) in anhydrous οβ-γΙοηοΙιΙοΓί ^ (6 µL) was added dropwise omeethyltrillnormfthanine nitrate (1.60 µL, 14.0 mmol) in a nitrogen tattooer.

The reaction mixture was allowed to warm to room temperature and stirred for 16 hours. Additional mercuric acid (0.40 mL, 3.56 mmol) was added and after 3 hours at room temperature the solvent was removed by suction.

The residual oil was dissolved in ether and the resulting gum-like material was dissolved in water (5 L). The solution was cooled in an ice bath and a solution of sodium hydroxide (844 mg, 21.1 mmol ·) in water (5 L) was added. After stirring for 4 minutes, the solution is diluted with 60 ml of water and the pH is adjusted to 8 by addition of solid primary potassium phosphate.

Then, 40 mL of this solution (about 4.7 mmol of a mixture of isomeric triazolium thiols) was added to an ice-cooled, stirred solution of enol phosphate (2.00 g, 3.45 mmol) in tetrahydrofuran (60 mL). The mixture was stirred in an ice bath for half an hour and then transferred to a pressure vessel containing a suspension of 10% palladium on carbon (2.00 g) in ether (60 mL).

The reaction mixture is hydrogenated for 1 hour at 50 psi. The organic phase was separated and washed with water (2 x 10 mL). The combined aqueous phases are filtered and the filtrate is concentrated under high vacuum (about 1 mm Hg, 1.5 hours).

The residual solution is then chromatographed (medium pressure reverse phase column, 45 x 130 mm, water as eluent) to give, after lyophilization, 595 mg of a mixture of isomeric carbapenems which are contaminated with a small amount of inorganic substances.

These were separated and purified by high pressure liquid chromatography (10 x 300 mm Waters Microbondapack C _1g column, multiple dosing, water as eluent) and obtained in order of elution: isomer B, 153 mg (13%);

¹ H NMR (D ₂ O) δ: 1.23 (3H, d, J = 6.4 Hz), 3.12 (2H, q, J = 1.4, 8.9 Hz), 3.39 (1H δ, J = 2.7, 6.0 Hz), 4.07-4.68 (10H, m), 8.19 (1H, s).

IR (nujol) 1755 cm.

UV (phosphate buffer, pH = 7.4, M = 0.05) .lambda.max: 296 nm (epsilon = 6.700); and isomer C; 284 mg (24%).

^{Χ ΗΝΜΡ} (D ₂ O) delta: 1.23 (3H, d, H = 6.4 Hz), 3.15 (2H, q, J = 3.7, 9.0 Hz), 3.37 (1H δ, J = 2.6, 6.0 Hz), 3.95-4.65 (10H, m), 8.62 (1H, s).

IR (nujol) 1750 cm

UV (phosphate buffer, pH 7.4, M = 0.05) λmax: 298 nm (epsilon = 7.600).

Example 16 (5R, 6S) -6- (1R-Hydroxyethyl) -3- (2-methyl-], 2,3-thiadiazolium-4-ylmethylthio) -7-oxo-1-azabicyclo [3.2.1] hept- 2-ene-2-carboxylate

OH

A. Ethyl 1,2,3-thiadiazol-4-ylcarboxylate ¹ z

CH ₃ ¹ CD Hurd and RI Moře, J. Am. Chem. Soc. , 77 o

II - NHCOEt 1 - T

II ^c x

COEt

II o

5359 (1955).

$ oci ₂

O

II

COEt

A solution of ethyl alpha-N-ethoxycarbonylhydrazone propionate (31.2 g, 0.154 mol) in thionyl chloride (80 mL) was stirred at 23 ° C for 3 hours and then heated at 70 ° C for 20 minutes. The thionyl chloride was evaporated and the residue was triturated in hexane (4 x 30 mL). The red solid was dissolved in dichloromethane (150 mL) and the solution was washed with saturated sodium bicarbonate solution and water.

After drying over sodium sulfate, the solution is concentrated until the compound begins to crystallize. He was allowed to stand at ²³ and the crystals are filtered, ^16, 8 g, mp ° C ⁸ 6 69%. The filtrate was concentrated and purified by chromatography on a silica gel column and eluting with dichloromethane. Yield 3.17 g, mp 86 ° C, 13%.

IR (KBr) gamma: 1720 (ester) cm max ^ MR (CDCl ^g) d: ¹ 52 (3 ^ t, J = ^7, 1 CH ^g CH ₂ ^{O) 4,} 57 (2 ^ ^q, J = 7 ^, 1 ^ ' ^,

CH ₃ CH ₂ O), 9.47 (1H, s, thiaphiazole H).

B. 1.2 TI-OH ^iso-1-4 - ¹ loetluno ^o1

Líaih ₄

Ramby ^, SO of SB Ross and NE Stjernstoom,

Acta Pharm. Sss ^ ^cl.ca,

1 (2285-96 (1973); C.A. 79, 137052W (19773)).

To a suspension of ethyl 1,2,3-thiaphiazol-4-ylcarboxylate (18.35 g, 0.116 mol) in ether (400 mL) was added lithium aluminum hydride (2.47 g, 0.065 moo) portionwise over 1 hour. The reaction mixture was stirred ^and Odin PTO 7 ^h 23 ° ^C and ^p of ^IFA tothummatominimhydrto ^{(2.47 g 0 0} 65 oo ^o).

Stirring was continued for 24 hours by the addition of water (7 mL), 15% sodium hydroxide solution (7 L) and water (21 mL). After stirring for 15 minutes, the ether solution was decanted and the gum was extracted with ether (5 x 100 to).

The ether extracts were combined, dried (MJSO 2) and (5.4 g). The crude material was purified on a silica gel column (120 g, 4 x 16 cm) and eluted with ether to give 1.3 g (7%) of ethyl 1,2,3-0-thiaphiazol-4-ylcarboxylic acid and 2.45 g ( 18%) 1,2,3-Otiafhazol-4-ylmethanols.

IR (µm) λm: 3380 (OH) cm. max. ¹ HMR (CDCl 3) δ 2 ^, 31 (1µs ^, OH) ^, 5 ^, 22 (5s ^, CH 2 O) ^, 8 ^, 50 (12s ^, H 2 O ^, azo 2).

C. 1,2 3-O-thafiazo-4-yl • methanol methanesulionate

MsCI

NEt ₃ ^>

CH ₂ OMs

A solution of 1,2,3-0-thiaphiazole-44 -44-methyl-methanols (0.75 g, 6.5 mmol) in dichloromethane (20 mL) was added in^andt^mooiéto d^atwith^íku cooled to⁵ °^C a ^^^idá se - ^ ietoylamto⁽1^{, 0}1⁸ ob^7.3 mmc> l) and o^E:^tan sulionyl chloride (0.565 ml, 7.3 ornoH)

After 15 minutes, remove the ice bath and stir the reaction for 2 hours. The solution was washed with 1N hydrochloric acid (2 x 2 ol) and water, dried (MgSO 2 + MgO) and concentrated. The residue was purified by chromatography on a silica gel column (1.5 x 21 cm) and eluted with ether to give 0.90 g (71%) of 1,2,3-triazol-4-ol.

IR (IMI °) ^G and ^M: 1350 (SOP cm ^"1, 1172 _(SO) cm '\ ¹ HMR (CDDip delta ^ ^3, 09 ^{(32, s,} C 2 ^{2, 5,} 75 ^(22, s ^, CH ² ), 8 ^, 72 (12) s ^, H Otilphenol · s) UV (2 2 (^) 12) and λ _max : 251 (epsilon 1990).

Analysis: calculated for C ₆ H ₆ N ₂ O ₃ S 24.7 3% C, 3 H 2 · 14.42? N, 3 3.02% S;

found 24.78 '·. N, 14.66% N, J 1, 94.4%

0.13 g (19%) of di- (1,2,3-thiadiazol-4-ylmethyl) ether,

IC (film) lambda _niax : 1,272, 1,242, 1,200, 986, 805, 728 cm ^-1 . ¹ H NMR (CDCl ₃ ) δ: 5.16 (s, 4H, CH ₂ ), 8.42 (s, 2H, thiadiazole H).

D. 4-Acetylthiomethyl-1,2,3-thiadiazole

CtyOMs q

II

CH ₃ CSNa

II

CH ₂ SCCH ₃

To a solution of 1,2,3'-thiadiazol-4-ylmethanol methanesulfonate (0.90 g, 4.6 mmol) in tetrahydrofuran (9 mL) was added an aqueous solution (2 mL) of sodium thioacetate (prepared from thiol acetic acid (0.38 mL)). (5.3 mmol) and sodium bicarbonate (0.445 g, 5.3 mmol).

The resulting mixture was stirred at 23 ° C for 1 h and diluted with ether (75 mL). The organic solution was washed with water (3 x 3 mL), dried (MgSO ₄ ) and concentrated. The crude mixture was purified by silica gel column chromatography (1.4 x 19 cm) and eluted with 50% ether in hexane to give 0.60 g (75%).

IR (film) gamma: 1675 . max Δ H NMR (CDCl 3) δ: 2.37 (C = O) cm ^-1 (3H, s, ch ₃ i 1, 4.58 (2H, s, _CH2), δ 8.44 (1H, s, thiadiazole H) Analysis: for ^C 5 ^H 6 ^N 2 ^OS 2 C, 34.47; C, 3.47 % H, 16.08 % N, 36.80% S; found 34.48% C, 3.83 % H, 16.28 % N, 36.80% S.

E. 4-acetylthiomethyl-2-methyl-1,2,3-thiadiazolium trifluoromethanesulfonate and 4-acetylthiomethyl-3-methyl-1,2,3-thiadiazolium trifluoromethanesulfonate

II

CH ₂ SCCH ₃

CF ₃ SO ₃ Me

O lí

CH ₂ SCCH ₃

N

CF3SO3

O

II

CH ₂ SCCH ₃ ^CF 3 ^SO 3

To a solution of 4-acetylthiomethyl-1,2,3-thiadiazole (0.60 g, 3.44 mmol) in a mixture of ether (4 mL) and dichloromethane (0.4 mL) was added several crystals of the compounds over 5 minutes. and trifluoromethanesulfonate (0.407 mL, 3.6 mmol).

The reaction mixture was stirred under nitrogen at 23 ° C for 6 hours. The white solid, which is a mixture of the two title compounds, was filtered and washed with ether. 1.05 g, 90%.

IR (KBr technique) gamma: 1675 (C = 0) cm, ^J max ^{1 H} NMR (DMSO, d = 6) δ: 2.43 (3H, s, CH ₃ COS), 3.33 (s, CH ₃ on N) -3), 4.57 (s, CH ₃

to N-2), 4.66 (2H, s, _CH2), 9.55 (H on thiadiazolium N-2), 9.66 (H on thiadiazolium N-3). Analysis: for C _? H ₉ N ₂ O ₄ S ₃ F ₃ calculated 20.27% C, 2.38% H, N, 9.45; S, 32.46; found 24.61% C, 2.57% H, N 8.47%, S. 28.21%

F. 4-Mercaptomethyl-2-methyl-1,2,3-thiadiazolium trifluoromethanesulfonate and 4-Mercaptomethyl-3-methyl-1,2,3-thiadiazolium trifluoromethanesulfonate

O

II

A solution of a mixture of 4-acetylthiomethyl-2-methyl-1,2,3-thiadiazolium trifluoromethanesulfonate and 4-acetylthiomethyl-3-methyl-1,2,3-thiadiazolium trifluoromethanesulfonate (1.05 g, 3.1 mmol) in 6N hydrochloric acid ( 10 ml) was heated to 65 ° C under a nitrogen atmosphere for 1.75 hours.

The solvent was evaporated under reduced pressure to give 0.91 g of a yellow syrup. This compound was used for the next step without purification.

G. (5R, 6S) -6- (1R-Hydroxyethyl) -3- (2-methyl-1,2,3-thiadiazolium-4-ylmethylthio) -7-oxo-1-azabicyclo-3.2. o] hept-2-ene-2-carboxylate

Cold solution (5 ° C) of (5R, 6S) paranitrobenzyl 6- (1R-hydroxyethyl) -3- (diphenylphosphono) -7-oxo-1-azabicyclo (3.2.0) hept-2-ene-2-carboxylate ( 1.7 g, 2.92 mmol) in tetrahydrofuran (10 mL) was treated with a solution of a crude mixture of 4-mercaptomethyl-2-methyl-1,2,3-thiadiazolium trifluoromethanesulfonate and 4-mercaptomethyl-3-methyl-1,2,2 3-thiadiazolium trifluoromethanesulfonate (0.9 g) in a mixture of phosphate buffer (pH 7.2, 0.3 M, 15 mL) and tetrahydrofuran (5 mL).

The reaction mixture was stirred for 1 hour and the pH was maintained at 7.2 with 2N sodium hydroxide solution. Stirring was continued for an additional hour and then ether (50 mL) and 10% palladium on carbon (1 g) were added. The resulting mixture was hydrogenated at 23 ° C for 2 hours and filtered through celite at 310 kPa.

The organic phase was separated, diluted with ether (50 mL) and phosphate buffer (pH 7.2, 0.3 M, 20 mL) and hydrogenated (2 g of 10% palladium on carbon) at 50 psi for 2 hours. The aqueous phases were combined (from the first and second hydrogenolysis), washed with ether and purified by prepPak 500-C / 18 chromatography, eluting with water to give 0.22 g of crude material.

This was re-purified by high pressure liquid chromatography and water as eluent to give 0.040 (4%) of the title compound after lyophilization.

IR (KBr) Gaman _χ: 3400 (br, OH), 1745 (C = 0 of beta-lactam), 1580 (carboxy ^la -t) ^cm3.

HMR · ^X (D2 O) delta: 1.23 (3 R ^'d, J = 6.3 ^ ^C ^ CHOH), ^{3.04, 3.05, 3,} 16 (21 ^{(п ъ} H-4), 3.38 (1H, dd, J = 2.8 Hz, J = 6.0 Hz, H-6), 3.9-4.6 (2H, m, H-5, CH-CHOH), 4 51, 4.53 (2s ^, SCH 2), 4.61 (s, N ⁺ CH ³ ).

UV (H_O) lambda 224 (epsilon 4,345), 262 (epsilon 4,980), 296 (epsilon 6885) q- zmax [alfaJ2 ⁵ 18 ° (c 0.18, H ^j O).

- 4o

T1 / 2 = 9.8 h (measured at a concentration of 10 M in phosphate buffer pH 7.4 at 36.8 C).

EXAMPLE 17 'l- [5- (1 -karboxylátomethyl N-methyl-l, 2,3-triazine ° ium) methanthio2 -6.alpha.-FL- (R) ^{1-hydroxyethyl] -7-oxo-azalňcykl.of} 3 ^2] o t ^h e ^{p-2-ene-2 -k} arboxylát potassium

O

II

HOC

1) LAH

2) MsCl / NEtg KSCOCH 3

O

II

BrCH2COEt

c) / ^Pd-C, Chan ^K m ^{p s} sus rnethyíL enz ⁱ ^ - ^^^ ^ from tr. ^^ arboxy.ové to ^{y if y} N £ c. ^ erson Acta

Chem. ^S cand. ^{13, 888} (1959)] (9 ^{00 g, 70.9} mmol ..) in ^b m ^s ezvod tetra- ^hy dro ^furan (200 mL) was added in small portions lithium aluminum hydride (2.83 g, 70.9 mmol ).

The mixture was stirred at room temperature for 15 hours, then 20% aqueous sodium hydroxide solution (20 mL) was cautiously added in approximately 1 mL portions. The resulting granular suspension was filtered and the solid was washed with additional tetrahydrofuran (5 x 75 mL). The combined tetrahydrofuran solutions were dried (MgSO 4) and the solvent was removed. The residual yellow oil is chromatographed on a silica gel column (90 x 35 mm), eluting with a 100 ml portion of hexane, ethyl acetate / hexane (1: 1) and (1: 3) and finally ethyl acetate and methanol (9: 1). . Obtained 4-hydroxymethyl-1-methyl-1,2,3-triazole (3.18 g, 40%) as a colorless oil.

¹ H NMR (CDCl ₃ ) δ: 4.07 (3H, s), 4.73 (2H, d), 7.52 (1H, s).

IC 3,320 cm ^-1 .

To an ice-cooled stirred solution of alcohol (4.67 mL, 41.3 mmol) and triethylamine (7.47 mL, 53.7 mmol) in methylene chloride (20 mL) was added dropwise methanesulfonyl chloride (3.82 mL, 49.6 mmol). . After 1/2 hour the solvent was removed and the residual solid was triturated in acetonitrile (30 mL).

Potassium thioacetate (7.06 g, 62.0 mmol) was then added and the suspension was stirred at room temperature for 3 hours. An additional amount of potassium thioacetate (3.0 g, 26.3 mmol) was added and the suspension was stirred for an additional 16 hours. The dark colored suspension was then concentrated and water (10 mL) was added.

The mixture was extracted with methylene chloride (5 x 40 mL). The combined extracts were dried (MgSO 4) and the solvent was removed. The residual oil was chromatographed on a silica gel column (90 x 36 mm), eluting with hexane followed by hexane / ethyl acetate (1: 1). There was thus obtained 4- (methanthiolacetate) -1-methyl-1,2,3-triazole (5.95 g, 84%) as a light pink solid.

¹ H NMR (CDCl ₃ ) δ: 2.40 (3H, s), 4.10 (3H, s), 4.20 (2H, s), 7.53 (1H, s).

IR (nujol) 1675 cm ^1st

A solution of triazole (1.00 g, 5.85 mmol) and ethyl bromoacetate (1.48 mL, 13.3 mmol) in anhydrous acetonitrile (10 mL) was heated at 60 ° C under nitrogen for 90 h. The solvent was removed and the residual oil was triturated with ether (4 x 25 mL) to give 1-methyl-3- (ethylcarboxymethyl) -4-methanthiolacetate-1,2,3-triazolium bromide as a brownish gum which was used directly for another reaction.

To an ice-cooled, stirred solution of triazolium bromide in water (20 mL) was added a cold solution of potassium hydroxide (0.66 g, 12 mmol) in water (5 mL). After 20 minutes, it is diluted to 35 ml and a quantity of solid primary potassium phosphate is added to adjust the pH of the solution to 8.0.

This solution was then added to a stirred ice-cooled solution of enol phosphate in tetrahydrofuran (35 mL). After 1/2 hour, the reaction mixture was transferred to a pressure vessel containing ether (35 mL) and 10% palladium on carbon (1.5 g).

The mixture is hydrogenated for 55 minutes at 50 psi. The organic phase was then separated and washed with water (2 x 5 mL). The combined aqueous phases are filtered and the filtrate is concentrated under high vacuum. The remaining sc was chromatographed on a reverse phase column (35 x 120 min) and eluted with water.

Lyophilization of the carbapenem-containing fractions yielded 1.20 g of a green solid. This was rechromatographed on Waters Prcp. 500 on a high pressure liquid (PrepPAK 500 / Cg) column and elute with 2% acotonitrile in water. Fractions containing carbapenem were pooled and lyophilized. This material was rechromatographed by high pressure liquid chromatography (10 x 30 mm Waters Microbondapack (18 column) and eluting with water to afford, after lyophilization, the pure title compound, 1 '> _H1 (17) as light yellow. solids.

1 H NMR (D ₂ O) δ: 1.24 (3H, d, J = 6.4 Hz), 3.07 (2H, d, J = 9 Hz), 3.38 (1H, q, J = 2) 7, 6.0 Hz), 4.02-4.30 (3H, m), 4.29 (3H, s), 5.23 (2H, s), 8.52 (1H, s).

IR (nujol) 1750 cm ³

UV (phosphate buffer, pH 7.4) .lambda.max: 296 nm (epsilon = 7.520).

Example 18

3- [4- (1-carboxylatomethyl-3-methyl-1,2,3-triazolium) methanthio] -6alpha- [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo [3.2.o] hept- Potassium 2-en-2-carboxylate

O

II

HOC-Ξ

Et fl CHOC

b) NaBH ₄ O II y-OC-N =)

P0 ₃

a) MeOTf

b) KOH

HSCCHI and ³

A mixture of ethyl azidoacetate (30.0 g, 0.23 mol) and propiolic acid (14.3 mL, 0.23 mol) in toluene (75 mL) was stirred at room temperature. The reaction is slightly exothermic and after 1 1/2 hour it rapidly becomes strongly exothermic, so ice bath cooling is required.

After the exothermic phase is complete, the reaction mixture is refluxed for half an hour. It is then cooled in an ice bath, the crystalline substance is filtered off and washed with a little toluene. The crude material thus obtained (33.3 g, 72%) consists of a single isomer.

¹ HMR ^(DMSO-dg) d: ¹ 20 (3 ^ t, J = 7 HZB ^4, 15 (2 ^ ^q, J = 7 HZB ⁵ 42 (2 ^ s) ⁸ 67 (1H, 3) , probably 1- (ethylcarboxymethyl) -1,2,3-triazole-4-carboxylic acid analogously to O. Pederson, Acta Ohem. Scand., 13, 888 (1959), cited previously.

A solution of this carboxylic acid (5.00 g, 25.1 mmol) and triethylamine (3.68 mL, 26.4 mmol) in anhydrous methylene chloride (50 mL) was added to an ice-cooled stirred solution of ethyl chloroformate (2.52 mL, 26 mL). , 4 mmol) in anhydrous methylene chloride (50 mL).

The purple solution was stirred for half an hour, then washed with water (10 mL), dried (IMJSO 3) and the solvent was removed. The crude mixed anhydride was dissolved in tetrahydrofuran (50 mL) and added slowly to an ice-cooled suspension of sodium borohydride (0.72 g, 18.9 mmol) in tetrahydrofuran (50 mL).

After stirring for half an hour, additional sodium borohydride (0.30 g, 7.9 mmol) was added and the reaction mixture was left in an ice bath for 1 hour. Water (5 ml) was added and after 10 minutes a 10% aqueous hydrochloric acid solution (3 ml) was added.

After gas evolution had ceased, solid potassium carbonate (2 g) was added with stirring. The organic phase was then removed and the remaining white paste was extracted with additional tetrahydrofuran. The combined organic phases were dried (MgSO 3) and the solvent was removed. Silica gel column chromatography eluting with hexane, ethyl acetate / hexane, and finally ethyl acetate gave 1- (ethylcarboxymethyl) -4-hydroxymethyl-1,2,3-triazole (2.04 g, 44%) as a crystalline solid .

¹ H NMR (CDOl ^) d: ¹ 28 (3 ^ t, J HH =), ^4, 23 (2 ^ 7 J HZB ^4, 75 (2-s)

4.85 (2H, s), 7.73 (1H, s).

To an ice-cooled solution of triphenylphosphine (5.47 g, 20.8 mmol) in anhydrous tetrahydrofuran (100 mL) under a nitrogen atmosphere was added dropwise diisopropylodicarboxlate (4.11 mL, 20.8 mmol). After half an hour, an ice-cooled solution of alcohol (1.93 g, 10.4 mmol) and thiol acetic acid (1.49 mL, 20.8 mmol) in anhydrous tetrahydrofuran (50 mL) was added under nitrogen atmosphere.

The mixture was left in an ice bath for 2 hours and then at room temperature for a further 12 hours, after which time the solvent was removed. The reaction mixture was chromatographed on silica gel (40 g, eluting with 100 mL portions of hexane, 5%, 10%, 15% ... 50% ethyl acetate and hexane).

Fractions containing thioacetate were combined and re-chromatographed on silica gel (60 g), eluting with 200 mL portions of hexane, 5%, 10%, 15%, 20% ethyl acetate and hexane, and 22.5%, 25%, 27.5%. 35% ethyl acetate and hexane.

1.24 g (49%) of 1- (ethylcarboxymethyl) -4-methanthiolicetate-1,2,3-triol are obtained as a crystalline solid [ ¹ H NMR δ: 1.28 (3H, t, J = 7 Hz) 2.37 (3H, s), 3.87 (2H, s), 3.90 (2H, q, J = 7Hz), 5.12 (2H, s), 7.63 (1H, s) .

IR (nujol), 1735, 1780 cm @ -1 and an additional 1.40 g of a triphenylphosphonic acid contaminated material.

To an ice-cooled stirred solution of this triazole (1.00 g, 4.12 mmol) in anhydrous methyleneiloride (5 mL) was added dropwise methyl trifluoromethanesulfonate (0.51 mL, 4.53 mmol).

The bath was removed after half an hour and after another half hour the solvent was removed in vacuo. A white solid was obtained, which was suspended in water (15 ml) and the stirred mixture was cooled in an ice bath. A solution of potassium hydroxide (0.69 g, 12.4 mmol) in water (5 mL) was added and the reaction mixture was stirred for 1 hour.

It is then diluted with 30 ml of water and solid primary potassium phosphate is added to adjust the pH to 8.0. Part of this solution (22 mL, about 3.0 mmol of thiol carboxylate) was added to an ice-cooled, stirred solution of enol phosphate (1.60 g, 2.76 mmol) in tetrahydrofuran (30 mL).

After half an hour, the reaction is complete and the tetrahydrofuran is removed under high vacuum. The yellow solution is then chromatographed on a reverse phase column (35 x 120 mm), eluting with water (300 mL) and then 100 mL portions of 5%, 10%, 15% .... 30% acetonitrile and water.

Lyophilization of the desired fractions gave the p-nitrobenzyl ester (930 mg) as a yellow solid. This was transferred to a pressure vessel containing ether (25 mL), tetrahydrofuran (25 mL) and a phosphate buffer of 2525 mL, prepared by dissolving primary potassium phosphate (1.36 g, 0.01 mol) in water (100 mL) and adjusted to pH to 7.4 by adding 45% aqueous potassium hydroxide solution] and 10% palladium on carbon (900 mg).

The reaction mixture was then hydrogenated at 50 psi for 1 hour, then the organic phase was separated and washed with water (2 x 5 mL). The combined aqueous phases are filtered and concentrated under high vacuum. The remaining solution is chromatographed on a reverse phase column (35 x 120 mm) and eluted with water.

Fractions containing carbapenem were combined and lyophilized to give 1.21 g of a light green solid. This was then purified by high pressure liquid chromatography (10 x 300 mm Microbondapack C-18 column) and eluted with water to give the pure title compound 480 mg (41%).

¹ H NMR (D ₂ O δ: 1.23 (3H, d, J = 6.4 Hz), 3.11 (2H, d, J = 9 Hz), 3.37 (1H, q, J = 3) 0.16 Hz, 4.02 (7H, m), 5.18 (2H, s), 8.53 (1H, s).

IC (nujol) 1750 cm 4

UV (phosphate buffer, pH 7.4) _Amax: 205 nm (epsilon = 7810).

Example 19

3- [5- (1,4-Dimethyl-1,2,4-triazolium) methanthio] -6alpha- [1- (R) hydroxyethyl] -7-oxo-1-azabicyclo [3.2.o] hept-2-ene -2-carboxylate

A. 1-Methyl-5-methanthiolacetate-1,2,4-triazole

(a) MsCl / NEt ₃ _______ β

b) ch ₃ csh / net ₃

O

To an ice-cooled stirred solution of 1-methyl-5-hydroxymethyl-1,2,4-triazole (565 mg, 5.0 mmol) [r. Jones and C. Ainsworth, J. Amer. Chem. Soc., 77, 1938 (1955)] and triethylamine (0.91 mL, 6.5 mmol) in methylene chloride (5 mL) were added dropwise methanesulfonyl chloride (0.46 mL, 6.0 mmol).

After 20 minutes, additional triethylamine (1.05 mL, 7.5 mmol) was added followed by thiol acetic acid (0.53 mL, 7.5 mmol) and stirring was continued for 45 minutes. The reaction mixture was then diluted with methylene chloride and washed with water.

The aqueous phase was extracted with methylene chloride (3 x 5 mL) and the combined organic phases were dried (MgSO ₄ ) and the solvent was removed. Silica gel column chromatography gave pure 1-methyl-5-methanthiolacetate-1,2,4-triazole (570 mg) as a yellow oil [further, the impure fraction (200 mg) was re-chromatographed on a thin layer of silica gel to give an additional 100 g. mg pure (total yield 85%).

¹ H NMR (CDCl ₃ ) δ: 2.38 (3H, s), 3.90 (3H, s), 4.25 (3H, s), 7.80 (1H, s).

b. 3- [5- (1,4-Dimethyl-1,2,4-triazolium) methanthio] -6alpha- [1- (R) -4-hydroxyethyl] -7-oxo-1-azabicyclo [3.2.o] hept -2-ene-2-carboxylate

a) MeOTf

b) NaOH

To an ice-cooled solution of 1-methyl-5-methanthioacetate-1,2,4-triazole (730 mg, 4.27 4.27 mmol) in methylene chloride (7 mL) was added dropwise methyl trifluoromethanesulfonate (1.20 mL, 10.7 mmol). ). The reaction mixture was allowed to warm slowly to room temperature over 3 hours and then concentrated. The residual oil was triturated with ether to give crude 1,4-dimethyl-5-methanthiolacetate-1,2,4-triazolium trifluoromethanesulfonate (1.46 g), which was used directly for the next reaction.

To an ice-cooled solution of the triazolium salt (1.45 g, 4.35 mmol) in water (5 mL) was added a solution of sodium hydride (512 mg, 12.8 mmol). After 45 minutes, the reaction mixture was diluted with 25 mL of water and the pH adjusted to 7.6 with solid primary potassium phosphate.

This solution was then added to an ice-cooled, stirred solution of enol phosphate (2.00 g, 3.45 mmol) in tetrahydrofuran (25 mL). After 30 minutes, the reaction mixture was transferred to a pressure vessel containing ether (40 mL) and 10% palladium on carbon (2.0 g).

The reaction mixture is then hydrogenated for 1 1/4 hour at 310 kPa. The reaction mixture was then diluted with ether (25 mL) and filtered. The organic phase was separated and washed with water (2 x 5 mL). The combined aqueous phases were washed with ether (3 x 25 mL) and then concentrated in vacuo.

Reversed phase column chromatography (45 x 130 mm, eluted with water) was followed by lyophilization of the carbapenem-containing fractions to give 650 mg of crude material. This was re-chromatographed to give the pure title product 450 mg, 39%.

¹ H NMR (D ₂ O): 1.24 (3H, d, J = 6.4 Hz), 3.19 (2H, q, J =, 2.6, 9.2 Hz), 3.45 (1H δ, J = 2.8, 6.0 Hz), 3.91 (3H, s), 4.06 (3H, s), 4.08-4.36 (2H, m), 4.54 ( 2H, d, J = 2.8 Hz), 8.71 (1H, s).

IR (nujol) 1755 cm

UV (phosphate buffer, pH 7.4) λ max = 294 nm (epsilon = 8.202).

T ₃ y ₂ (phosphate buffer, pH 7.4, M = 0.067, T = 37 ° C) 9.1 h.

(1'R, 5R, 6S) -3-1,3,3-dimethyl-5-tetrazolimethylmethyl-thio] -6- (1-hydrooxytthyl-7-oxo-1-azabicyclo [2.2.1 hept-4-en] - carboxylate

Example 20

A. 5-thiocyclohexyl-22-methacrylate and 5-ethoxycyclohexyl-methacrylate

la. Meehylation of palzomethane.

A solution of 5-methoxycyclohexyl ether (9.17 g, 0.064 mol D. D. Moodrhack, Chem. Bee., 108, 887 (1975) in ethyl ether (80 mL) using a mixture of ethanol and ether) gave the same isomer ratio cooled to 0 ° C and a solution of diazomethane (3 g, 0.071 mol) in ether (200 mL) was added dropwise over 15 minutes, the light yellow solution was stirred for 30 minutes and the excess diazomethane was quenched by the addition of acetic acid (1 mL).

Evaporation of the solvent and distillation of the residue gave a clear oil, mp 95-100 ° C / 66 Pa, 9.64 g, (96% yield). The nuclear maagonic resonance spectrum shows that the ratio of 1-methyl. The 2-methylisomers in the mixture are 6: 4. Roodděení these two isomers can not be accomplished by distillation or HPLC CH roi ^ ^r ^ o MTC RFF: i. * iC (film) by ^ ax! 1,740 cm ”· · (C = 0 ester);

^X HMR (CDC1 ³⁾ delta: 1.53 (3H, disodium ^p řekrytí t, J = ^7, 0, CH ₂ CH _3), 4.46 and 4.53 (2S 3 ^S, CH 3-one and 2 mthyl- methyl ethylrthole, 6: 4 ratio; the 2-methylisomer is in the lower field and is the product in the minimum amount of 4.5 ppm (2H, double overlap q, CH 2 CH 2).

lb. 5-tert-cyclohexyl-2-methyltetradole

A mixture of 5-ethoxycarbonyl-2-methyltetraethyl and 5'-hexyloxybutyl-methyltetrazole. (0.252 g, 1.61 1.61 mmol, 1: 1 ratio of both isomers) in iodomethane (0.5 mL) was sealed in a glass tube and heated at 100 ° C for 15 hours and at 130 ° C for 6 hours. Desalting reaction blanket. to give the title compound as a pale yellow oil: 0.139 g (55%, bp 95-100 ° C / 1 mm Hg).

iC (film) _ax : 1740 cm ”(C = O ester).

¹ HMR (CDC1 ³ delta ^{1 4} 6 (3-t, J = 7.0, CH ^{- CC2), 4,} 53 (3H, s, CH ^ b ^4, 5 (2H, ^q, J = 7, O, CH 2 Cl 2 (H 3).

2. Methylation with dimethyl sulfate

A solution of 5-ethoxycarbonyltetrazole (1.42 g, 0.01 mol) in anhydrous acetone) (20 mL) was treated with anhydrous potassium carbonate (1.38, 0.01 mol) and dimethyl sulfate (1.26 g, 0.01 mol). ). The mixture was heated to reflux for 12 hours. The carbonate was filtered off and the solvent was evaporated under reduced pressure.

The residue was diluted with dichloromethane (30 mL), washed with saturated sodium bicarbonate solution (10 mL), brine (10 mL), and dried over anhydrous sodium sulfate. Evaporating the ^{p STARTUP} DLA and rain ^and lac ⁱ va in the CIS ^to uu ^{ka or} rice-ol ^{1.45 g} (9% ^3); TV ^{85 to 10} 0 C ^{/ 66} and ^P. Nuclear magnetic resonance indicated the presence of two isomers in a ratio of 1: 1.

B. 5-Hydroxymethyl-2-methyltetrazole

1. Reduction of the ester mixture

A mixture of 5-methyltetrazole and ethoxykarbony1-1-5-carbethoxy-2-methyltetrazole (ratio ^6: 4) (7. ^{60 g,} 0. 9 0 ⁴ mol) in anhydrous tetrahydro ^{furan yd} ro ⁽⁵ ° mL) oc ^h la ^DI at 0 ° C and to keep ^and to react with lithium borohydride (1.06 g, 0.049 mmol) in small portions over 15 minutes. The mixture ^E is u ^{d d} ržuje al ^{IC h 30} m nu ^{i t at 10} ° C for ^t and ^p OM ^s m CH ^{and 4 h} o ^d ins ^{instance 20} ° C.

After och ^l AD ^and at ⁰ ° C and ^for creating excessive ^{hyd y} te ri ^d u o ^{p l} atm ^of extended ^runs of ^adding it to ^{6N ky y L} IN hydrochloric acid (at pH 7 has no more gas evolved). The solvent was evaporated in vacuo and the residual oil was diluted with dichloromethane (200 mL), washed with brine (10 mL) and finally dried over sodium sulfate.

Evaporation of the solvent and distillation of the residue in vacuo gave 1.83 g (33%) of a clear oil. The nuclear magnetic resonance spectrum shows that the product is 5-hydroxymethyl-2-methyltetrazole.

2. Reduction of 5-ethoxycarbonyl-2-methyltetrazole

To a solution of 5-ethoxycarbonyl-2-methylterazolu (0,13'9 g, 0.89 mmol) obtained by isomerizing a mixture ^of an ester ^with methyl iodide in anhydrous ^{HYL dé} m ^t Etra ^hyd ro ^furan (1 ^mL) was s l ⁰ BC ^IDs and ^p evný lithium borohydride (0.019 g, 0.87 mmol).

The mixture was allowed to warm slowly to room temperature and stirred for 4 hours. Excess borohydride was extended on ^p ^ Gd atrným Pr ^I given ^r M N ^{6 alkyl} with ^y c ^l in ^{the hl} orovodí b ^s at ⁰ ° ^{C), pH 7).} The ^solvent was evaporated and the residue was dissolved in dichloromethane (25 mL) and dried over anhydrous sodium sulfate.

Evaporation of the solvent gave the title compound as a clear oil: 0.092 g (91%), TV from 90 to 120 ° C / 66 Pa (dec.) ^IR (film) ^n, Y _max: ^{3350 cm -1} (Shiro ^{to ý OH)} .

¹ HMR (CDCl ^j) d: ^4, 4 (2 ^ ^s, CH _{β -2Ь} ^4, 93 (2 R ^s, CH ^).

C. 5-acetylmercaptomethyl-2-methylterazole

1) Μ8α.Ε1 ₃ Ν »-у _с „ _гОН I л- / Λ * сн ₃ CH ₃

To a solution of 5-hydroxymethyl-2-methyltetrazole (1.83 g, 11.7 mmol) in anhydrous dichlormetoanu (25 mL) at ⁰ ° C ^{was added} metliansuhon BC ^ C ^ orm ^{(1, 4, 7 g,} 12, ⁹ mmoll then ^run em minutes was added dropwise triethylamine (1.30 g, 12.9 mmol).

Sm^Es se m^andC^{há 1} totenu^{at 0} ° C a^pthen the necM reacted with the solution^toem^thioacetá.tu^dpeat (1.60 g, 14.0 mmol) in anhydrous Ν, Ν-dimethylformamide (10 mL). The resulting gel was stirred 3^hO^din^y pH⁰ Deň: 32 ° C.^Rea^CZKn^and the mixture is from^ŘE^dí (hclúormetoanem⁽2 °⁰ ml^{, p}rom^yis with a muzzle c^hlO sodium chloride (20 mL) and dried over anhydrous sodium sulfate.

Evaporation of the solvent in vacuo and chromatography of the resulting oil on silica gel (2 x 15 cm (2 x 15 cm, eluting with dichloromethane / dichloromethane and acetone 5%) gave the title compound as a clear oil: 1.31 g (65%).

IR (film), the hind ^_X 1696 cm ^-1 (C = O thiosteru) ¹ HMR (CDCl · ³⁾ d ^e LTA: ² 43 (3H, ^s, SAc), ^4, 36 (3H, s, 2-CH ^{3), 4} 38 ^{ppm (2-a,} _5-CH2).

D. ^{5-mercaptomethyl-y} l-l, 3-dimethyl ltetrazoliumtrifluorrnethansulfonát ^y / CH3

Г CH ² SAc ^С ^ ° 3 ^СН 3> © \ - _{CH SH}

X ©

QF ₃ SO ₃

CH3

A solution of 5-acetylmercaptomethyl-2-methyltetrazole (0.400 g, 2.32 mmol) in anhydrous dimíchá chloromethane (3ml) was treated with trifluormethylmethansulfonátem (0.76 g, 4.64 mmol.) And 16 ^h of ^di n ^at 22 Noc: 2 ° C.

Evaporation of the solvent in vacuo gave a red oil. This salt-free water O (5 mL) and 4M sodium hydroxide (0.8 ml, 3.2 40 mi.nut ^p s ^ ^{0, R} E from with ^DI water (7 mL) and saturated sodium Baptist? O ² The resulting clear solution was kept under nitrogen and used directly to dissolve in mmol). The mixture was at pH rav ^{d p} 7, ^{the third} for the next stage.

cold stir

E. (l'R, 5R, 6S) -3- / l, 3- dimethyl-5 ^{d-tetrazolium)} methylthio / -6- (1-hydroxyethyl) -7-oxo-l-

IR (KBr technique) (C = 0 beta-lactam) ^, 1 max tu).

N-bicyclic (3.2.0) hept-2-ene-2-carboxulate

(br, ^{even years, for} C = O R ^b oxy ^{la- 1 R} HM (D? d ^{and e LT:} 1.24 ^(3H, d, J = ^6, 4 ^Hz, C H HOH), ^3, 0-3 ^, 3 ⁽² ^ m, H-4), ^3.4 (1H, dd, J = 5.8, J = 2.9, H-6), 4-4.2 (2H, m, H-5 and OH 2 OHOH), 4.34 and 4.5.7 - (2 x - ¹ -?

CHg-1 and 3 tetrazole), 4.49 and 4.51 (2H, 2s, CH 3 S).

The product has a half life of 10.5 hours at 37 ° C (c = 10 ~ ⁴ M in pH 7.4 phosphate buffer.

Example 21

Alternative procedure for the preparation of 3- (N-methylpyridin-2-ylmethanthio) -6alpha- [1- (R) -hydroxyethyl] -7- oxo-1-azabicyclo/3.2.0/- hept-2-ene-2-carboxylate

OH

In a 2 liter flask equipped with a Vigreaux distillation column for magnetic distillation, a heating mantle and a nitrogen inlet was added 4.0 mol (432 mL) of methyl acetoacetate and 8.0 mol (464.6 g) of allyl alcohol.

The reaction mixture was distilled at 92 ° C for 12 hours. 136 ml (2.0 mol) of allyl alcohol are then added and the mixture is distilled for 23 hours. 136 ml (2.0 mol) of allyl alcohol are then added and the mixture is distilled for 16 hours. The reaction mixture is then distilled under vacuum and the product is collected at 105-110 ° C (50 mm Hg) to give 414 g of allylacetoacetate (73% yield).

To a solution of allylacetoacetate (226.5 g, 1.594 mol) in 3 liters of acetonitrile and triethylamine (243.4 mL, 1.753 mol) was added p-toluenesulfonylazide (345.3 mL, 1.753 mol) over 1 hour while cooling bath temperature. maintained at about 20 ° C.

The reaction mixture turned yellow. The reaction mixture was then stirred under a nitrogen atmosphere at room temperature for 18 hours. The mixture is concentrated on a rotary evaporator. The residue was dissolved in diethyl ether (2.6 L) and 1M aqueous potassium hydroxide solution (800 mL). The organic phase was washed five times with 1M potassium hydroxide solution (500 ml) and once with brine (400 ml).

After drying over magnesium sulfate and concentration on a rotary evaporator (temperature -30 ° C), 260.2 g (97%) of the title compound are then obtained.

To a stirred suspension of allydiazoacetoacetate (203 g, 1.195 mol) in 2 liters of methylene chloride and 199 ml (1.444 mol) of triethylamine at 5 ° C was added 302 ml (1.315 mol) of tert-butyldimethylsilyltriflate over 45 minutes. , '

The mixture was stirred at 5 ° C for 1 hour and then for 1 hour without cooling. The reaction mixture was washed with water (4 x 500 ml) and then with brine (500 ml). Thereafter, 344 g of an orange oil are obtained, dried over sodium sulfate and concentrated. This oil was used directly for the next step.

D.

I i

, ососн ₃

To a mixture of (1R, 3R, 4r) -3- (1'-tert-Butyldimethylsilyl-oxyethyl) -4-actoxyazetidin-2-one (28.7 g, 0.1 mol) and freshly melted zinc chloride (6 8 g (0.05 mol) in anhydrous methylene chloride (700 ml) was added dropwise a solution of allyl-2-diazo-3-tert-butyldimethylsilyloxy-4-butenoate (33.84 g, 0.1 mol) in methylene chloride over 5 hours. (50 mL).

The mixture was stirred at room temperature for 2 hours, after which a small amount of starting material remained. An additional amount of allyl-2-diazo-3-tert-butyldimethylsilyloxy-4-butenoate (4.23 g, 0.015 mol) in 10 mL of methylene chloride was added over 1 hour and stirring was continued at room temperature for 10 hours.

The reaction mixture was then diluted with ethyl acetate (750 mL), washed (2 x 300 mL saturated brine), dried over magnesium sulfate and evaporated to give 62.5 g of a dark orange oil, which was dissolved in methanol (500 mL) and combined with 1N aqueous hydrochloric acid and stirred for an additional 2 hours.

The reaction mixture was concentrated to half volume and poured into a mixture of ethyl acetate (800 mL) and water (800 mL), and the combined aqueous extracts were washed with brine (2 x 400 mL), dried over magnesium sulfate and concentrated.

32 g of a dark orange-red oil are obtained. Chromatography gave 9.33 g (33% yield) of the title compound as a golden yellow oil which solidified in a pale yellow solid.

¹ HMR (CDCl.) D ^{6, 20-5,} 72 (ль 2H), ^{5, 48-5,} 21 2H), 4.74 (dt, J = 5 ^ J '= ^1, 2 Hz,

2H), 4.30-3.88 (m, 2H), 3.30-3.20 (m, 2HI, 2.89 (dd, J = 7.3, J '= 2.1, 1H), 2.18 (s, 1H); 1.32 (d, J = 6.2, 3H).

A mixture of the alpha-diazoester prepared according to Step D above (9.2 g, 32.7 mmol) and rhodium acetate Rh (OAc)) in benzene (1 L) was refluxed for 1 hour. The solution was treated with activated carbon and filtered through celite.

This was then washed with 100 ml of hot benzene. Concentration of the filtrate afforded 8.08 (97% yield) of the title compound as a light brown crystalline solid.

¹ H-NMR (CDC- ²⁾ d ^{6, 15-5,} 68 (m, 1H), 5-5-5,18 (m, ^{2H), 4, 71-4,} 60 (m, ^2H); 4.40-4.05 (m, 2H), 3.17 (dd, J = 7.1, J '= 2.0, 1H), 2.95 (dd, J = 6.9, J' = 18.9, 1H), 2.42 (dd, J = 7.6, J '= 18.8, 1H), 1.88 (s, 1H), 1.39 (d, J = 6.3, 3H) ).

The mites on ^the ketoes ^t ERU ^{EXAMPLE IP} Raven ^eh o ^p e ^dl of growth ^{of E p} n (7, ^{5 g, 0.03.} Honey) ^{at 0} ° C under nitrogen was added dSSyopropylamin (6.08 ml , 0.035 mol) and then diphenylphosphoryl chloride. After 15 minutes, thin layer chromatography indicated that the mixture no longer contained any starting material.

Disisopropylamine (6.26 mL, 0.036 mol) and a solution of freshly distilled 2-mercaptomethylpyridine (4.5 g, 0.036 mol) in 5 mL acetonitrile were added to the reaction mixture. After stirring for 2 it ^di- n ^Y m ^s c ^{h and at 0} ° C and the mixture on ^l eje ^d of et ^{hyl -acetate} in (1 HRT), ^p rom ^y is water (2 x 150 mL), saturated sodium hydrooenuhličitanu solution (150 mL), water (150 ml) and brine ⁽² 00 ml).

The organic phase is dried over magnesium sulphate and concentrated to a dark orange-yellow gum. Chromatography gave the product as a golden yellow oil. This genus ^{p k} t ^p is extended breath ^{as well as} in (úethytotoeru and oc ^hl and ^says to ⁰ ° C.

Filtration gave 4.8 g (44% yield) of pure title compound as cream colored crystals.

^X H-NMR (CDCl ^g) d ^{8, 6-8,} 4 (m, 1H), (m, 2H), 4.80 to 4.66 (m, 2HU,

1.32 (d, J = 6.2,

^7, 85-7,15 (m, 3H), ^{6, 2} 0-5,74 (m, ^{1H), 5, 54-5,} 15

4.29-4.03 (m, 1H), 4.19 (s, 2H), 3.69-2.85 (m, 1H), 2.97 (s, 1H),

G.

To a solution of the allyl ester prepared according to Step F (1.79 g, 4.97 mmol), tetrakistriphenylphosphine palladium (175 mg, 0.15 mmol) and triphenylphosphine (175 mg, 0.67 mmol) in methylene chloride (25 mL) was added solution 2. potassium ethylhexanoate (1.085 g, 5.96 mmol) in ethyl acetate (12 mL).

The mixture was stirred at room temperature for 1 hour, then only traces of starting material could be detected by thin layer chromatography. The reaction mixture was diluted with anhydrous diethyl ether (150 mL) and the precipitate was filtered off, washed with ethyl acetate and then ether to give a light brown solid. This material was dissolved in water (10 mL) and purified by reverse phase chromatography to give 1.85 g of the title compound as a cream colored solid.

This was further purified by trituration in acetone to give 1.47 g (83% yield) of pure title compound.

^X H-NMR (D ₂ O) delta: ^8,45-8, 36 (m, 1HB ^{7, 92-7,} 22 (s, 2H), 3.34 to 2.71 (m, 3H), 1, 19 (d, J 3 H).

compounds

^4, 69 (m, 3H), ^{4, 78-3,} 91 m, 2H);

H.

^-3-k ar ^b o ^-

To matting Suy enz ^{i p} (° ^C) 6 ^h y ^d roxyethy -fŽ L ^ p ^ r ^ y ^ e ^ y ^^ ^ artiapenem of potassium azodicarboxylate (53.8 mg, 0.15 mmol) in acetone (2 mL) was added toluenesulfonic acid (2 ⁷ 6 п ^{0, 0,} 16 mmol ^{1). R} e and ^{d at the} CN is a mixture ^of m ^I C ^{h and 20} mktá ^{at 0} ° ^C and ^p o ^t om ^{added methylcarbazole IFL t} tr (0.02 ml). The mixture was stirred at 0 ° C for 60 min before LA-1 resin was added followed by hexane (6 mL). The reaction mixture was extracted with water (4 x 0.5 mL) and the combined aqueous phase was purified by high pressure liquid ¹ by reverse phase chromatography to give 10 mg of the title compound.

Example 22

Preparation of 3-N-methylpyridin-2-ylmethanothio) -6alpha-1- (R) -hydroxyethyl-7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylate by a one-pot procedure .

CH ₃ CN

0 °, 45 min

O

C1P (O0) ₂

EtN (iPr) ₂

NaOH

H ₂ O, 0 ° C, 1h

COjpNB

OH

A. Preparation of enol phosphate (2J

OH OH Дг T—— H П Tz = ° - -Ν ^ / ^ ^{0Ρ, 0} ^ 2 Qr COjpNB í CO ₂ pNB To January cooled solution ketone £ (3 g, 8.62 mmol) in acetonitrile (30 mL) was added

ethyldiito-propylamine (9%, 1.04 equivalents, 1.57 ml - addition time about 2 minutes) and chlorodiphenylphosphate (9 mmol, 1.04 equivalents, 1.87 nl - addition time about 2 minutes).

The reaction mixture was stirred for 45 minutes and then the disappearance of the ketone 8 was detected by thin layer chromatography (ethyl acetate, silica gel). The solution was diluted with ethyl acetate (60 mL), washed with cold water (2 x 50 mL) and brine, dried over sodium sulfate and concentrated ^etting (warm-ta ^la Zn ^{E = 20} ° ^C) of ^{i SKK} transmit ^{the to} TERA used as such.

B. Preparation of thiol (£)

1) NaOH / H ₂ O ° C, 1 h

---

2) KH2PO4

To a January cooled solution of thioacetate 3. (3.31 g, 10 nmol, in water purged with nitrogen for 5 minutes) was added dropwise (about 5 minutes) a cooled solution of sodium hydroxide (1.75 equivalents, 17.5 nmol, 0.7 g) in water. water (8 mL).

The reaction mixture turned yellow. After 75 minutes under a nitrogen atmosphere, the pH was adjusted to 7.4 with a saturated aqueous solution of primary potassium phosphate. The reaction mixture was diluted with water (15 mL). An aqueous solution of thiol δ (50 µl, 0.2 mmol / ml) was used as such.

C. Coupling

OH £ ° thf-h ₂ at pH 6.5-7.5 -OF T 0 °, 1 hr COjpNB

To the cooled solution of intermediate 2 (crude, prepared in A, 8.62 nmol) in tetrahydrofuran (50 ml) was added dropwise an aqueous solution of the thiol 8 prepared in B (5 ml of solution every 5 minutes).

100 ALIGN!

During the reaction, the pH of the reaction mixture is maintained between 6.5 and 7.5 (preferably 7) by the addition of cooled sodium hydroxide solution. The progress of the reaction was monitored by thin layer chromatography:

(a) silica gel, ethyl acetate;

(b) reverse phase. Analtech RPSF, CH 2 N, pH buffer (4: 6).

At the end, 1.15 equivalents of thiol (50 ml solution) was used. The reaction is completed after 1 hour ^at 0 ° C and the mixture oužije ^p j ^{and k} of ^the network ^T and ^P ro hy ^d by hydrogenation ^IP ou ^p Raven ^pH Ί.

D. Hydrogenation

CO ₂ pNB

The reaction mixture containing the intermediate (s ^{p p} Raven under paragraph C) was transferred to a Paar flask containing tetrahydrofuran (10 ml), phosphate buffer), pH 7, 0.1 M) (10 mL), ether (75 mL) and 10% palladium on carbon) (5 g) and hydrogenated at 310 kPa at ^{3 to 10} ° C for 2 hours.

The catalyst was then filtered off, washed with water (3 x 100 ml) and the pH was carefully adjusted to 6.2 with cold 2N sodium hydroxide solution. Ether was added, the aqueous phase was separated and washed again with ether.

The aqueous phase is rinsed under vacuum with an organic solvent and then purified on a Bondapack C-18 column (100 g, 4.5 x 1 cm cm) with cold distilled water. The pale yellow product-containing fractions (detected by UV and thin layer chromatography) were lyophilized to give 1.46 g (50% yield calculated from the bicyclic ketone) of product 6 as a yellow powder. Lambda 293, epsilon = 9,000, lambda 271, epsilon = 11,064.

Example 23^Ppreparation (51 (6 ^ -3- ^ [м 3- (Methy ^ y ^^ n ^ m ^ -y¹⁾ met^bullfinch thdo] -6- [l^{- (}RJ^-hy^droxyethy J -7-oxc-^l-azabicyyk^lO^TO3^,2.6ΐ | he^pt-²-en-²-kar^box^yl(²3A) (41 (51 (6S) -3-j2 [(^l, i-dumethylpyr-L-n-m-⁴[methyl] thio]^{-6- [}1- (R) -hydroxyeth^ylj-4-methy¹-7-oxo-^l-and for^biC^yto^lO¹3^,2^,Oj he^pt-2-en-²-karbox^yland^tat⁽²3B)

3B К - сн ₃

101

A. Preparation of 4-hydroxymethyl-3-methylpyridine

1) H ₂ O ₂

------------ 7>

2) _Ас э 0

3) н ₃ о ®

A solution of 3,4-lutidine ° C and ^p Otom the straight-APE

To prepare HY (froxymeetlhУpyyidiuu pouuije the general procedure of Booekeheida (V. Boekelheide, WJ Liuu JACS, 7 6, 1 286 (1954)). Freshly destioovaný ^{(4 6} 3 0 ^{g 0.4} moles) in 1 mL of ² ° ice βθΙ ^ oc ^ ^{1 t} b ^s is cooled to 0 64 ml of 30% hydrogen peroxide.

Vvrdklý ^The organic solution is heated for ³ him <din ^s at ⁷⁵ ° C to about ^ Jové Užni.

Pa ^{to be} at additional ml of 30% hydrogen peroxide and heating was continued for 18 hours. Nalkonec added again is ^{TE 20} ml ^{30% p} Erox ^id for reatání for cables and the mixture was maintained for an additional ^{3 ohu-1} at 75 ° C.

The solution was then bends to about 100 ml, was added ^and nu s ^l volume. The resulting mixture was cooled ⁽ 0-5 ° C) and 40% aqueous sodium hydroxide solution.

with 50 ml of water and quenched to about pH ¹⁰ using sclerotin.

The mixture was then extracted with methylene chloride (5X) and the extract was dried (Na 2+ + Na 2 SO ₄ ) in a yellow solution. Dilute this solution with hexane to dry in vacuo. There was obtained 3,4-lutene-N- oxide of the substance.

and concentrated on a rotary evaporator to give a solid which was filtered off and (48.0 g, 83%) in an off-white solid

N-oxide was added portionwise to 60 ml of acetic anhydride and the resulting dark orange solution was warm-in-A in ^d her ^La Zn ^and at about 9 ⁰ DEG ^C for a period jt ^d N ^{e H} o <din ^{y. For} the excess acetic anhydride is about ^dd ftSilujt under reduced pressure to give substance boiling at 90-120 ° C ^{/ 13} 3 Pa ^(39, 0 g).

Chroomtoogrfii this oil (silica gel / ethylacefát-petroleum ether 2: 3) afforded the pure ^{(19.0 g, 30%)} in the ^f orm ^of an oil: IR (oil) 1745 ^cm.

To 100 ml of 10% aqueous hydrochloric acid solution was added acetates and heated to boiling is nu ^d Todina. The reaction mixture was cooled to 0 ° C and quenched with solid sodium chloride and then extracted with methylene chloride (3 x 100 mL).

Or ^g Annie extract was washed with brine, dried over sodium sulfate and evaporated under VZN ^ in ^{11.0 g of} off ^of bhe ^{solid. t} . ^t . ^{Mp 70-72} ° C. ^{p-resisting substance} and triturated with cold ether to give pure 4-3-hydroxymethhУ mettlr Уppridiu (9.5 g, 67%) as a white ^{substance P} and EVN ^{and t. t} .

485 (1972) m.p.

⁷⁷ to ⁸⁰ ° C (according to ^W. LF Armare ^g o ^, b and SC S armor ^, JCS ^,

81-82 ° C).

HNMR (CDC13)

4.67 (s delta: 8.27, 7.41 (ABq, J = 5 Hz, 2H), 8.18 (s, 1H), 5.63 (br s, -OH,

CH ₂ ), 2.20 (s, CH ₃ ); I ^Č (nujol) 3,170 ^c m ~ ¹ .

B. Preparation of 4- (acetylmethyl) -3-methylpyridine

102

To an ice-cooled stirring solution of a mechanical stirrer trOfenulfosfinu (17.04 g, 0.065 mol) in 250 mL of anh ^ ého d00slpropulaнod0karboxulát tetrahydrofuran was added dropwise ^(12, 8 m ^{1 0.065} Mou and ^IKL MMN and the suspension ball ^ ^{at 0} ° ^C jetdnu todm .

To this mixture was added dropwise a solution of 4-iodomethyl-3-methulpuridine (4.0 g, 0.0325 mol) in 100 mL of anhydrous tetrahydrofuran. and then fresh distilled acetic acid (4.64 mL, 0.065 mol).

^The Zn ball ^Ikla mixture ^ ^{at 0} ° ^C and then ^d nu nu ^d is todhu ^at rt ^{t h} e ^{pl of} bowls as ^{to the t.}

to give an orange solution. The solution was concentrated on a rotary evaporator and then diluted with petroleum ether. The mixture was filtered and the filtrate was evaporated to give an orange oil.

Chromatography (silica gel / hexane and then 10% 50% ethyl acetate / hexane) of this oil yielded 7.0 g of an oil which was distilled (spherical condenser) to give the pure product (6.0 g, 100%) as a yellow oil, tv (air bath temperature) 95-100 ° O / 13.3 Pa.

³ 1H NMR ^(CDCl3) delta: ^{8, 40, 7, 2} 0 (AB ^q, J = 5 ^{Hн, 2H),} 8. 37 (s, 1H), ^4, 08 OH ²⁾

^{2, 35-CH ₃₎ 2} OH 32 ^j b IR (oil) 1695 cm ^'first

O. Preparation of 4-acetylthóomethyl) ^-1,3-d dOmetiulpurO OnOumtriflulrmethansulfonátu

CH ₂ SAc OH 3

CH ₂ SAc ^{1 / ch} 3

To an ice cooled solution of the thioacetate (2.95 g, 0.016 mol) in 10 mL of p metiulencilor0du to ^Rik, and ^p e meth ^IUL tr ⁱ f ^{l h} ulrmet ansulfon ^{carboxylate (4.60} m ^{1, 0.04} moto and mixture m ^s c ^{ha at about} 0 under nitrogen for 1 hour.

The reaction mixture was then evaporated to dryness and the residue was triturated in ether. The resulting solid was filtered off and dried in vacuo. Obtained ( ^4.0 g, 72%) as a white solid. ³ 1H NMR _(CDCl3) .DELTA: ^8, 72 (s, 1H), ^8, 58th ^7, 87 (AB ^ J = 6Hz, 2H), ^4, 39 ^N-OH-4, 17 (s, ^OH-2, 53 ^{(s, OH-2, 36-OH} _3); IR (oil 1 700 cm ^-1 .,

D. Preparation of (5R, 6S) - 3 - (£ (1,3- ^d 0Inetiylpyri ^d inium-4-yl) methyl thio I) - 6- [1- (R) -hydroxy-yethyl - 7-LXL-l- azaboyl 1,3,2 of hept-4-ene-4-carboxylate

CH ₂ SAc

Л \ ^сн 3

OH

2)

CO2PNB

Pd / O

OH

CH

0 '

103

An ice-cooled nitrogen purged solution of sodium hydroxide (0.324 g, 0.008 mol) in ¹⁰ mL ^{of 1} as ^{always when} gives ^th ioacetát (1 ^ ⁰ дъ ^0.004 mo ^l) and the mixture m ^I CH ^{and at 0} ° C VA ^tm osfé ^of and nitrogen for 1 hour. .

The pH was then adjusted to 7.2-7.3 with a 10% aqueous potassium dihydrogen phosphate solution and the resulting solution was added dropwise to an ice-cooled solution of enol phosphate (1.45 g, 0.0025 mol) in 20 mL of tetrahydrofuran.

After ^Ch m ^{at 0} ° C for one Iiothnu and ^p is ^p Otom Renesmé to t.akové to ^ tjy. K t ^{s and} the mixture is added 20 ml of ether, 25 ml of 0.1M phosphate buffer (pH 7.4) and 1.4 g of 10% palladium on carbon. The mixture is then hydrogenated at 310 kPa for one hour.

The reaction mixture was filtered through celite and washed with ether and phosphate buffer pH 7.4. The aqueous phase was separated and the remaining solvents were removed in vacuo. The resulting aqueous solution was applied to a reverse phase column (C18 g of Bonda ^p ak) which was effused with water and then with 10% acetonitrile in water.

Lyophilization of the appropriate fractions gave 0.9 g of an orange solid. This was rechromatographed using water and then 2% acetonitrile in water as eluent. Lyophilization gave pure compound 23A (0.25 g, 57%) as a yellow solid.

¹ H NMR (D?) Delta: 8.55 (s, ^1H), 8. ^53, 7.96 (AB ^q, J = ^6, 8 ^Hz, 2H), ^{4, 30-3,} 99 (m, 2H) 4.27 (s, 5H), 3.35 (dd, R. = 2.8 Hz, J = 6.0 Hz, 1H), 3.05 (d, J = 8.8 Hz, 2H), 2 50 ^{(s, 3H),} 1. 23 ^{(^,} J = ^6, 3 ^{Hz, 3H,} IR (KBr) ^1755, 1590 ^{cm -1;} UV ^(p UFR phosphate, ^p H 7)

295 nm (ejpsilon 7,180).

E. P^řprava (41 ^ 5R, 6S) -3-^[| U^{, 3-di}met^hylpyr^idin^andum-4-y¹ ) -methy 1^] thi-6-fluoro-4H-hexoxy ethyl] -4-methyl-4-oxo-L-azabicyclo [3.2.0] hept-2-ene-2-carboxylate

O

Pd / C

To an ice-cooled solution of an i.e ketone bicycle (0.906 g, 0.0025 mol) in 10 mL acetonitrile was added diphenylchlorophosphate (0.544 mL, 0.00263 mol), diisorpoyplethylanine (0.457 mL, 0.00263 mol) and 4-dimethylaminopyridine sequentially. (0.3 mg).

After 50 minutes with cold ethyl acetate and then wash with cold water and brine. ·. > ιΌ ·. The phases were dried (Na 2 SO 4) and evaporated at room temperature to give a yellow solid. '<, even in ·. '. The foams are foamed.

This foam is &quot; melt &quot; () ml. il. yd iofu r.-i: u, cool to -30 ° C under a nitrogen atmosphere or thereafter: - water / in rozi 4. · -m (l (i) cd-iodine made from 0.3 g of sodium hydroxide (II) (ii) - (ii) (ii) mm (ii) .-dy.

^R 104 ea n ^{s CZK} mixture MIC ^h and at ^-30 ° C ^{for 30} minutes :, ^p o ^t om ^BC at ⁰ ° C for ⁷⁵ minutes, and finally transferred to a pressure vessel containing 20 ml of ether, 30 ml 0 M phosphate buffer pH 7.4) and 1.5 g of 10% palladium on carbon.

After hydrogenation at 310 kPa for one hour, the mixture was filtered through celite and the aqueous phase separated and concentrated in vacuo. The resulting solution was applied to a reverse phase column (C) (Bondapak) which was eluted with water.

Lyophilization of the appropriate fractions gave 1.2 g of a yellow solid. This was re-chromatographed (eluting with water to 4% acetonitrile in water) to afford, after lyophilization, pure 23B (0.020 g, 28%) as a yellow solid.

¹ HNMR (I-O) d ⁸ 53 1 H) ^^ 8 7.81 (AB ^ J = ^6, 2 2 H) ^{4, 38-3,} 98 (4H ^ n)

4.27 (s, 3H), 3.49-3.18 (rn, 2H), 2.51 (s, 3H), 1.25 (d, J = 6.7 Hz, 3H), 1.16 (d, J = 7.6Hz, 3H); IC (KBr) 1750, 1595 cm; UV (phosphate buffer, pH 7) 292 nm (epsilon 7,930).

Example 24 rava ^or (^ ^{R, 6S)} - ^{3- [[(1,2} - ^d ime ^th y ^lp yri-ammonium ^{di-4-yl) l-methylamino]} thio] -6- [1- ^(R) - hydroxyoxy-7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylate (24A); and

⁽⁴ ^ ^{5 R,} 6 ^S) ~ ³ ~ ^[[(1,2-di: methylene ^lpY Ri ^di n ⁱ um ^- 4-yl) methyl] -6- fl thic ^ ^{1 (R) ethyl-hydroxy-d} roxy ^4- Methyl-7-oxo-1-azabicyclo [3.2.0] hept-2-enecarboxylate (24B)

CH3

С ^О 2СНз

A. Preparation of ^4- Hydroxymethyl-4'-methylpyricidine

To a suspension of 95% lithium aluminum hydride (2.4 g, 0.06 mol) was added a solution of methyl 2-methylisonicotinate (14.0 g, 0.093 mol, P. Rumpb Bu ^11. Soc. ^Chi m. Fr ^ ⁶⁴⁸ ( ¹⁹⁵⁴ )). ) in ⁵⁰ ml o ^b EFTA ^DEH 150 ml of anhydrous ether was prepared according to O. Efimovsky et ERU ^{h at} - ⁵ ° ^C under nitrogen, e ^{F R} e nitrogen.

The mixture was stirred at room temperature for 30 minutes and then refluxed for 2 hours. An additional 1.2 g (0.03 mol) of lithium aluminum hydride was added portionwise and refluxed for one hour.

The reaction mixture was then cooled to 0 ° C and 3.75 ml of water, 3.75 ml of a 15% aqueous sodium hydroxide solution were added, followed by 11.25 ml of water.

The suspension is then filtered and the filter cake is washed with ether and then with ethyl acetate. The filtrate was evaporated to give a dark yellow oil which was taken up in acetonitrile and then filtered through a silica gel column (eluting with acetonitrile and then acetone).

^{September KA} is ^P ro ^d u ^k t (b # ^{7 g, 67%)} as ^f Orme ^F lu ^Teh of oil. ^{) H NMR} (CDCl @) ^{d t} el and ^8, 3 ^ ^7, 1O (AB q, J = 5 Hz, 2H), 7.17 (s, III), 5.42 (s, -OH), 4, 70 (s, CH 2), 2.50 (s, CH 3).

105

B. Preparation of 4- (aceeyl thif) ethyl) -2-methylpyridine

To a solution of triphenylphosphine (31.4 g, 0.12 mol) in 200 mL anhydrous tetrahydrofuran under nitrogen, ^fe re ^d u ^p us ^{Cart Rf} -5 ^o C. ^EXAMPLES APE (^ ^ azo ^ sojDrop artoxyHt ^(23-mL, 0Д2 mol) and the mixture was ball ^H and ^{P F} of -5 ° ^C is ^{d df} nu nu him.

To the resulting suspension was added a solution of 4-hydroxymethyl-2 ^- methylpyridine (7.60 g, 0.062 mol) and freshly distilled thiol acetic acid (8.60 mL, 0.12 mol) in 40 mL of anhydrous tetrahydrofuran over about 10 minutes.

^R e and ^{d at the} CN is a mixture ^of m ^d c m ^HA 3 ° ^{F FS T} nu ^{0 O} C ^p and ^the one it nu ^{p df Rf} te ^pl about ^{the T} m ^s society. The ^resulting suspension was filtered and the filtrate was concentrated to give an orange-yellow liquid, which was diluted with ether and filtered.

The filtrate was evaporated and the residual oil was chromatographed (silica gel / ethyl acetate-hexane 1: 1) to give thioacetate (8.87 g, 79.4 ') as a yellow oil.

^X H NMR (CDCl ^j) delta: ^8, 45, ^7, 03 (AB q, J = ^{5Hz, 2H),} 7. 08 (s, 1H), 4 ^ 4 (s, ^CH2), 2. 55 (s, CH 3 B ^2, 39 (s, ^CH3); IR (oil) 1.695 cm ^-1.

C. Preparation of 4-Mercaptomethyl-2-methylpyridine

To 15 ml of ice-cooled nitrogen purged 1N sodium hydroxide was added at once 2---acetylthiomethyl methylpyrfdin BC ^f 0 ^o C ^to rea ^d Hand śirasā ^p ROMs ether (hydrohalic acid and extracted with methylene chloride (3 (1,358 g, 0.0075 mol). After stirring for 15 min x5 ml), neutralized with concentrated ^LF We observe cHorovox 10 mL).

Evaporation of the methylene chloride solution of the oil, which gradually turns pink on standing.

afforded the thiol (0.89 g, 96%) as a yellow ^X H NMR (CDCl ^j) delta: ^{8, -3, 7,} 37 (AB q, J = 5 ^{Hz, 2H),} 7. 43 ^(s, 1H) ^3, 63 (d, JC ^), 2.55 (s, _CH3), 1.81 (t, J = 7.5 Hz, SH).

^7, 5 Hz

D. P ^Rip Rav and ^LL y ^| - ^{(5 R,} 6S) ^- 3 ^{- [[(2-meth yl} pyridin ^f d ^f n ^-4 - ^y lúmethyj TH ^F o | -6 ^- p (R) ^-H ydroxyeth ^y 1 | - ^7-oxo l-aza ^bf c ^y klo ^[ 3, ^2, θ '] he ^p t- ^2- en- ² - ^k ar ^b ox ^ylate

O'

106

To a January cooled solution of the bicyclic ketone (0.760 g, 0.03 mol) in 8 µl acetonitrile was added successively diphenylchlorophosphate (0.653 µl, 0.00315 mol), diisopropylethylamine (0.550 mL, 0.00315 mol) and dimethylaminopyridine (0.8 g) ).

Sn^Es se m^andch^and one^hO^dinu p^Řand⁰ ° C and pa^to se oc^hland^dí on^-2° °^C and^adds se ^ merka ^ o methyl 2-methylpyridine (0.620 g, 0.00446 mol) followed by diiso^ppyridylhyanine (0.550 µl, 0.00315 mol).

Reaction ^mixture to ndcM ^1.5 hours at ^-2 ^ ins 0 ° C and ^p Oton allowed ^and heated to te ^pl ol necessity. The resulting mixture was diluted with 50 mL of ethyl acetate, washed with water, saturated sodium bicarbonate solution and saturated ammonium chloride solution, dried over sodium sulfate and evaporated.

The remaining material was chromatographed on silica (eluting with ethyl acetate and then acetonitrile) to give a solid which was triturated in ether to give the pure product (0.820 g, 73%) as a white solid.

¹ 1H NMR _(CDCl3) delta: ^{8, 45, 7,} 09 (AB ^q, J = 5 ^Hz, 2Hb ^7, 15 (s, 1H), ^{'6, 25-5,} 80 (m, 1H), 5, 60-5.20 (m, 2H), 4.82-4.68 (m, 2H), 4.55-4.05 (n, 2H), 3.97 (s, 2H), 3.16- 2.93 (m, 3H);

^2, 55 (s, 3НЬ ¹ 84 br s, 1H), 1. ³² J = 6 Hz, ^3H); IR (oil) ^1777, 1695 Ci ^ ^1.

E. Preparation of (5 ^{R, 6} S) ^- 3 ^- [[(^^ ^ pyridiniun dimethoxy-4-yl) -methyl | thi] ^- 6- [1 (R) -Hydroxyethyl] ^-

-7-oxo- • l-azabicyclo ^[3, 2,0] hept-2-ene N-carboxylate OH in CH, Y IQÍ oZ IQ C0 ₂ ^e CH ₃

A solution of allyl ester (0.350 g, 0.936 mMol) in 6 NL anhydrous acetonitrile was cooled to ^-5 ° C and ^p when ^added methyl ^t h ^{y | t} rif ^about ucrmet ^h antu ^l sulfonate (Ο, ΜΙ ^n-3 0.98 ппю ⁰ ). ^{P 15} = CH Nin solution tetrakit (triphenyl phosphine) palladium (0.027 g, 2.5 mol%) and triphenylphosphine (0.027 g).

The reaction mixture was stirred for 5 minutes and then pyrrolidine (0.082 mL, 0.983 mmol) was added dropwise. A solid began to slowly precipitate from the resulting brown solution. SNES intensive M ^I C ^{h and 20} n ^and nu ^{t eg 0} ° C, then enclosing ^BC and ^D, and ¹⁵ ш ¹ stodenéta ⁽⁰ ° C) acetone AVM ^{Ch ck} was continued for ²⁰ m ⁱ nu ^t at ⁰ Deň: 32 ° C.

The resulting suspension was filtered and the residue washed with cold acetone and dried in vacuo to give 0.345 g of a beige powder. This material was taken up in a small amount of phosphate buffer pH 7 (0.05M) and transferred to a short reverse phase column (c? G Bondapak).

Elution with water and lyophilization of the appropriate fractions gave 0.255 g of a pale yellow solid. This material was rechromatographed as before to afford (after filtration) pure 24A (0.195 g, 60%) as a pale yellow solid.

¹ HNMR (I-O) d ^{8, 85, 7,} 83 (AB q, J = 6.4 Hz, 2НЬ ^7, 87 (s, 1H), ^{4, 32-3,} 95 (m, 2Η) 4, 22 (s, 2H), 4.17 (s, 3H), 3.32 (dd, J = 2.6 Hz, J ₂ = 6.1 Hz, 1H), 3.06-2.93 (m ^2H), 2. 74 ^(s, 3 ^ ^1, 22 (d, J = 6.4 ^Hz, 3H); IR (KBr) ^1757, 1590 cm ^-1; uv (phosphate buffer @ pH 7.4) 296 nm (epsilon 7,746).

F. executions ^llyl - ⁽⁴ R-5R-6S) - ³ - ^ff (2-met. ^Hylp yri ^d in ^- 4 ^- yl) ^methyl]] thio ^{j -} 6- ^(l- (R) -h ^yd roxyethyl 4-Methyl-7-cxo-O-azabicyclo [3.2.0] hept-2-ene-2-carboxylate

107

A solution of the alpha-diazoester (1.50 g, 0.00508 mol) in 12 mL of ethyl acetate and hexane (3: 1) was heated under gentle reflux under a nitrogen atmosphere, then 0.020 g of rhodium octanoate was added in one portion.

Nitrogen evolves rapidly for about 5 minutes and after boiling for a further 10 minutes the reaction is complete (thin layer chromatography). The solvents were then removed under reduced pressure and the remaining gum was taken up in 15 ml of acetonitrile.

^The organic solution was oc ^{hl DI} at ^- 5 ° ^C and not read rea ^g OVA ^T ^ fenyktáortostátem ^(1, 10 m ^1,

0.00533 mol), diisopropylethylaninen (0.927 ml, 0.00533 mol) and ^4-y dinethylaninop ridίnem (0.6 mg, 0.1 mol%).

Reaction ^mixture to m ^{d d} CHA, nu totúnu ^{at 0} ° ^C and then ortho position-to ^-2 ° ^C and NEC ^IIa is treated with a solution of 4-mer ^to aptonethyl- ² -neth ^ylpyridine (0.656 g, 0, 00533 mol) in 1 ml acetonitrile and then with 0.927 ml (0.00533 mol) disisopropylethylamine.

It was originated ^and m ^'C and ^{1.5 h} it ^di ny p ^s -l ⁰ ° ^C was ^{added p} Otom ^d and ^LSI TMO. ¹ (0.06 ^{6 (0,} 0.53 mmol) and dšisopropylethylamin (0.093 mL, 0.53 mmol). The reaction mixture was allowed to warm up over a n ^{d s h i} n ^g from about ¹⁰ ° ^C and ^p is Otom dilute ^{says 75} m ^one hundred ^d en ^Star by e ^t hy ^{1 -acetate} in ^p rom ^y Mou and ^extensible st ch ¹ or ⁱ du sočného) in ^y su ^width (magnesium Sa ^ YMB and ^dp and ^R ^ EPDO ^{Mp 30} ° ^C) .

The resulting gum was chromatographed on silica gel. Elution with ethyl acetate removed impurities and subsequent elution with acetonitrile gave the product (1.04 g, 53%) as a light yellow foam.

¹ 1 HNMR (CDCl ^g) deltéi: ⁸ 4 ³ 7.07 (AB q, J = 5 Hz, 2H), 7.10 1H), ^{6, 2,} 0-5,75 (m, 1H),

5.51-5.29 (m, 2H); 4.81-4.69 (m, 2H); 4.29-4.03 (m, 2H); 3.96 (s, 2H); 35-3.05 (m, 2H).

2.53 (s, 3HU, 2.16 (broad s, 1H), 1.33 (cd, J = 6.3 Hz, 3H), 1.22 (d, J = 7.3 Hz, 3H);

IR (o1e ^j) 1770, 1705 CIF ^first

G. rava at ^p (4R, 5R, 6S) -3-H (1, ² - ^{dimethylphenyl y1py} ri ^{d i} in m-4- ^y1) metoyf ^'t h ⁱ o! ^-6- [1- (R) -hydroxyethyl] -4-methyl-7-oxo ^{Y1 -} bazaMc ^ o ^{[з, 2 hept-2- ene-2-1AT} at ^the arboxy

WITH

108

A solution of allyl ester (0.582 g, 0.0015 mol) in 15 mL of anhydrous acetonitrile was treated under nitrogen with metiultrOfluormetiаnsulflnátem (0.178 mL, 1.575 mmol) at ^-5 ° ^{O. P 15} m ^ UTA ^ is p ^Rida roztoJí totoaJás ^ Rhen ^ tosf ^ PAH ^ a (0 ^ ³⁵ g. ² mo ^1%) and trifenulflsfόnu (0.035 g) in 1 mL methylenc ^ C ^ ode and, after 5 0.131 mL (1.575 mmol) of purr10done was added in minutes.

MMN ^Ikla mixture MIC ^{h and 20 t} mine ^at 0 ° ^C was ^added and the bow ³⁰ m ¹ STUDÉNKA (0-∞) of acetone. The mixture ^{E is in some of} ntenн m ^s c ^HA 15 mi N? ^At 0 ° ^C, and ^p and ^{k d} f sraž.enba about ^{0 l Truji,} promj ^t smoked with acetone and drying in the ^ Kuu afforded 0.520 g of beige powder.

Dilution of the filtrate with ether gave an additional 0.041 g of crude product. The combined solids were dissolved in a small amount of phosphate buffer pH 7.4 (0.05M) and loaded onto a reverse phase column (C ^ g Bondapak).

Elution with water followed by 2% acetonitrile and water gave, after isolation, 24B (0.413 g, 76%) as a yellow solid.

¹ H NMR ((3, 0) delta: ^8, 55 ^7, 76 (ABC ^ J 6? 3 ^{Hн, 2H),} 7. 81 ^(s, 1H), ^{4, 4-3,} 7 (m, 2H )

4.19 (s, 2H), 4.16 (s, 3H), 3.47-3.14 (m, 2H), 2.73 (s, 3H), 1.24 (d, J = 6, 4 Hz, 3H), 1.16 (d, J = 7.3 Hz, 3H); IR (KBr) 1750, 1595 cm @ -1; UV (phosphate buffer, pH 7.4) 293 nm (epsilon 7 170).

Example 25 ^Prep Rav (5R, 65) ^-3-one, O-chloride chmettyh ^ um ^ ² - ^) mettytoluo] -6- (R- ^ -hydroxy ^ t ^ l) metiul 4R-7-oxo- -l-azabicyclo ^to bicyclo [3,2,0] hept-ene-carboxylate -

A. Trifluromethanesulfonate salt (1,6-dimethylampurodon-2-ul) methultol

MeOTf ether

HOI, 6N.

Δ

CH ₃

A solution of (6-methylpyridin-2-yl) methulti0lαeetátu (1.0 g, 5.52 mmol) in anhydrous ether (5 mL) was treated under nitrogen with metiultrOfluormetiansulfonátem (0.74 mL, ^6.5 mmol ¹⁾ and m ⁱ C ^{the HA 4 and the} LD n p ^{s at 23} ° ^O.

The ether was decanted and the white solid was prlmuзe twice with ether (2 mL) and dissolved in an atmosphere with ^CART dus to ⁷⁰ ° ^C .and then zatosh for SNP / enélio ^{t), and} with at ^{Ps and} the UTA ru ^p. Stop hydrochloric acid by distillation with water (2 x 10 mL).

The crude material was purified by reverse phase chromatography on a reverse phase column (2.2 x 13.0 cm, PrepPak O-18) with water as the olefin solvent. Appropriate fractions were combined and ^ o ^ tion of a white powder, 1.43 g, 85.4%, iO (KBr) ^{n ymax:} 2565 (sh), 1626 (pyridinium), 1585 (pyridinium) cm - UV (λmax) λ max: 278 (epsilon 7355);

109

Calc'd for, N0, S2F, ^C 35.64, H 3.99, N 4.62, S 21.14 Found: C 35.49, H 4.05, N 4.56, S 20, 99.

B. ^(5R, 6S) ^{-3- [(M d d} imC'thylpyyi inium- ² - ^yl) methyl t ^h iq] ^- 6- ^(1R - ^{hydroxyacetophenone Y} et ^Hy1 U- ^4R- lip ^Hy1 - ⁷ ~

-ox3- ¹ -azabic ^YKi Q ^[3,2, O ³ hept-2 ^- ene-2 ^- carboxylic ^b ox ^y repent

O

1- (PhO) ₂ PC1

2- No (iPr) ₂

3- Thiol

4-Net (iPr) 2

H _2, Pd / C Buffer 7.0 ether

THF

To a cold (5 ° C) solution of ^(5R, 6S) п ^ ro ^ ^b enz ^y lj ^^ - DOCx-C l] ^Rh hydrox ^y e ^Thy1) -4 ^R S ^-

-meth^hl-¹-and for^biC^yk1Q £^3,2^, O]^heptan-2-^R-kar^box^ylatu (1.1.1 g^, 3,^{06 /} mmm^O, R / S:^86/14) in Icezvo Deacletoniril (90 ml) was added simultaneously with diphenylchlorophosphate (0.68 µm, 3.3 µmL) and n-propylethylamine (0.57 µl, 3.3 µmol) in a nitrogen atomizer over 10 minutes.

Wells (5 ° C) stirred nu ^d is one hour, cooled to ^-30 ° C and ^run 15 minutes eo ^p RID ^and simultaneously trfllurrmethansulfonátové salt solution (1.6 ^{direthhlpylidiniur-22-y} 1) reehhlthio1i (1.03 g, 3.4 mmol) in anhydrous acetoriniline (2 mL) and dlisopropylethylamine (0.59 µm, 3.4 mmol).

Vzzňklá mixture was stirred for about ^{0.5 h <di} ny p ^{s "30} ° C, warmed to ⁰ ° C к and ^p o dilute the cold water (35 ol) was stirred for one hour. The resulting emulsion is poured onto the top of a reverse phase column (PrepPak C-18, 2.5 x 18 cm) and then eluted with a mixture of 25-50%. in the water.

L ^y yUflinací appropriate fractions yielded a sticky yellow solid, 1.69 g, which was dissolved in wet tetarhydrofuranu (40 RL). To the solution was added ether (70 µL), potassium dihydrogen phosphate buffer and sodium hydroxide buffer (pH 7.0, 0.2 M, 50 mL) and 10% palladium on carbon ( ^1.6 9 ^g ) and the resulting mixture was hydrogenated ^{for 2 h hy} Odin p ^{s 290 kp} aa ²³ ° C., ^and filtered through otoo Cdit.

Two F ^and that άεΐ ί ^ ^ and the aqueous ^{one stage} of the slop etterer ⁽² x ^20mL) and curved with straight ^{L-C 233} C under high vacuum to 15 ml and applied on top of a reverse phase column (PrepPAK C- 18).

Elution with 4% acetonitrile in water yielded 0.23 g of the title compound mixed with diphenylphosphate potassium carbonate (24% roo) after liophilization of the appropriate fractions.

By further purification on a reverse phase column (2.5 x 14 cm, PrepPak C-18) with water (400 ml) and a salt of 10% and 50% strength in water (200 ml) ) as eluent, after člnideer of the relevant fractions gave a yellow powder, 0.17 g, 15.3% IR (KBr) _x Nym.: 1750 (C = O beta - 1aktar) ^{1625 P} yndiniur) 1 ^{600 (} C = ^O, N-oxyl; cm @ -1;

¹ H NMR (D ~ O) ^{d 1,} 1E, J = ^7, 2 Hz, CH at C ^ ^{-4, 1,} J = 24 ^6, 4 Hz, ² CIPCHOHb ⁸⁰

Z 3 (s, CH, on C-6 pyridinium), 4.18 (CH, on N pyyidinia), 4.41 (center of AB quartet, CH 2 S), 7.5-8.4 (hydrogens on pyridine) ); UV (buffer 0.05 M, pH 7.0) lambda _χ: 278 (epsilon 11504);

D -256 ^3, 4 (c ^0, 2 ^2, H ^{2 O),} tai ^{j / 2} = ^{2 0,} 8 h measured BC ^{1 37 3} C ^P i ^{f ru) p} H ^7, 4) Ex. endsetzrac 10 ^3S M.

110

Example 26

Preparation of (5R, 6S) -3- ^{[(1,6-DIMETHYLPYRIDINIUM} · 2-yl) methyl thičj --6- (lR-hydroxyethyl) ^{-7-oxo-l-azahicyklo [3, 2, 0] h} ept- 2 ^- ene-2-carboxylic ^yl methacrylate

CH ₃

A. Three! LuormctHéinsullonatová and (Hfenylíosfátová salt (5R, 6S) útrobenz ^ -b ^ ^Q1 6-dimotkyl. ^Py ri. ^{Of di-ammonium-2-yl)} MMT ^hy l Hno] -6 - ^{(i R} - ^hy droxyet ^hy l) - 7-oxo-aza R ^b ic ^YKL [3,2, θ | ^h ept-2-ene-2-carboxylate

MeOTf —7 ether

1- NaOH. /

ΚΗΏΓΟ4

CH3

OH 1- (PhO) 2 POCl ^CH 3 | ~ 2- Nee (iPr) 2 1> = o / 3- hУiel ČOOpNB 4- Nee (iPr) 2

coopNe

To the cold (⁵ ° C) Pitch (^5R,^6S)^p-ni.tro^benz^yl ~⁶- (^lR-^hydrox^yy^Eh^yyů-3^{, 7}-^dioxo-l -azabicyclo 3,2,0 heptane-2R-carbox^y(2.14 g, 6.14 mmol) in anhydrous acetonitrile (18 mL) was added diphenylchlorophosphate (1.37 µl, 6.6 mmO1), diisopropylethylamine (1.15 mL, 6 mL) in an atmosphere of nitrogen. 6 omo1, dilsopropylethylanine (1.15 mL, 6.6 mmO) such that it was^plota maintains^por 5 ° C (⁷ and^of 10 minutes) and 6-dimethylaminopyridine (6 mg,^0.05 mnm ^.

^MES was stirred for ^1.5 hours at ⁵ ° ^Y C ^{P F} OUZ is ^t and ^{the network,} the I ^ka. For the next ' ^p ' this mixture is called solution A '. A solution of (6-netУylpyridin-2-y1) neehyllhieacetátl (1.23 g, 6.8 mMol) in anhydrous ether (10 mL) under nitrogen atmosphere, treated with mehyMr ^ ^f ifluormeehansul He ^and tmii ^{(0, 85>} п ^(7.5 nunml) and OIC ^hook laodiny ^1.5 at ²³ ° C.

The ether was decanted off and the white powder was washed twice with ether (2 x 10 mL) and resuspended in water ( ²⁰ µl). Vvni ^kl ý aqueous solution was ocřiladí at 0 ° C free of ahnoefé ^R e ^s u ^phorous acid is reacted with sodium hydroxide (4N, 3.4 mL, 13.6 nnnol.

^The MES is níciaá nu ^d ^ pyrrolidine at ² ° C and then the ID ^{and BC} ^ nin d.il drogenfos potassium ^sulfate with the pH adjusted with ¹ to ^7.6 This mixture is for another ¹ označovdna procedure as extended ^S-B herein ^d with ⁽⁵ ° ^C) solution A was 0.5 hours to the solution B with dropwise addition of a 4N solution of

1 L sodium hydroxide maintains a pH between 7.25 and 7.35. The mixture was stirred for 0.5 h and poured onto the top of a reverse phase column (4.0 x 18 cm, Prep-Pak C-18). The column was eluted with 25-50% acetonitrile in water.

Lyophilization of the appropriate fractions gave the title compound in a), 80%; IR (KBr): 3700-3-1T (pyridinium) cm; HNMR (DMSO, a yellow powder)

000 (OH), 1772 (C = 0d-6) .delta .: 1.15 (d,

2.82 g (51% (PhO) ₂ PO ₂ , 49%, CF ₃ SO ₃ ester), 1,625 (pyridinium), 1,590

J = 6.2 Hz, CH ₃ CHOH), 2.84 (s, CH ₃ on C-6 pyridinium), 4.16 (s, CH ₃ on N pyridinium), 4.79 (s, SCH ₂ ), 6.6-7.5 (PhO) ₂ PO ₂ , 7.5-8.7 (pyridinium hydrogens and pNB ester hydrogens).

B. (5R, 6S) 3 - [] 1,6-dimethylpyridinium-2-yl) methylthio-6- (1R-hydroxyethyl) -7-oxo-1-azabicyclo [з, 2, 6] hept-2-ene -2-carboxylate

CF 2 SO 4 - ^J (PhO) ₂ PO ₂

COOpNB

H ₂ , Pd / C buffer

To a solution of trifluoromethanesulfonate and diphenylphosphate salt (49:51) (5R, 6S) p-nitrobenzyl-3- (1,6-dimethylpyridinium-2-yl) methylthio -6- (1R) -hydroxyethyl) -7-oxo-1- azabicyclo P, 2, (J Hept-2-ene-2-carboxylate, (0.87 g, 1.27 mmol) in wet tetrahydrofuran (50 mL) was added ether (50 mL), potassium dihydrogen phosphate buffer, and sodium hydroxide (0.1 M, 40 mL, pH 7.0) and 10% palladium on carbon (0.87 g).

The mixture was hydrogenated for 2 hours at 23 ° C at 10 psi and filtered through Celite.

The two phases were separated and the aqueous phase was washed with ether (2 x 15 ml), concentrated under high vacuum to 30 ml and poured onto the top of a reverse phase column (PrepPak C-18, 2.2 x 13 cm).

The column is eluted with water. Appropriate fractions were combined and allyophilized to give a yellow powder, 0.179 g, 40%; IR (KBr) ^{n ýRiax: 1755} (C = 0 of beta-lactam), 1628 (pyridinium), 1590 C = O carboxylate) cm ^{"1; 1} H NMR (D 2 O) δ: 1.25 (d, J = 6.4 Hz, CH 2 CH 2), 2.82 (s, CH ₃ na to C-6 pyridinium), 3.12 ('dd', J = 9.2 Hz, J = 2.9 Hz, H-4), 3.39 (dd, J = 6.0 Hz, J = 2.8 Hz H-6), 3.7-4.4 (CH ₃ CHOH, H-5, CH ₃ on N pyridinium), 4.48 (s, CH ₂ S), 7.6-8.4 (hydrogen on pyridinium); UV (Н _Э ) lambda. 279 (epsilon 9 628) with inflex at 296; I- ³ 55.0 ° (c 0.63,

H ₂ O); tau _3/2 = 12.5 h measured at 37 ° C in buffer pH 7.4 to a concentration of 10 M.

Example 27

Preparation of 3- {-2- (N-methylpyridinium) methanthiol-6alpha- [1 (R) -hydroxyethyl] -4-beta-methyl-7-oxo-1-azabicyclo (3.2.0) hept-2-cn-2 -carboxylate

112

NB

A. p-nitrobenzyl-2-diazo-3-oxo-n-valerate (4)

A solution of 50 g (0.35M) of ethyl 3-oxo-n-valerate and 54 g (° 35M) of p-nitrobenzyl alcohol in 400 ml toluene at ^{HRI 130-140} V ^{and 18 di-} n ^b ez ^PE TNE ^h o ch ^l a ^dce . ^Dp Arene ^m dispersant solvent ^ gave a yellow crystalline solid which was recrystallized from ether-pentane to give ⁷⁵ g ^(86% above ^{Photogene, white} e ^k) ^ nkroloenzybl-oxo-N-vatototu ^(3), mp 33-3 ⁴ ° ^C .

IR (KBr) ^g and ^m and 1,740 and 1,705 cm ^-1; NMR (CDCl ^{j) and delta:} 1.20 (3H, t, J = ^7, H ^from 0 b 2.65 (2H, q, J = 7.0 Hz), 3.60 (2H, s), 5 28 (2H, s), 7.45 (2H, d, J = 9.5 Hz) and 8.18 (2H, d, J = 9.5 Hz).

^The mites ^to in ^{55.5 g (°} 22M) with the amine compound act ^{l f} ny ³ in ⁵⁰⁰ m ^l acetoiňtrHu ^FS is 0 ° ^{C 45 g} of p ^RFDA (0,44м) of TEA and then 50 g (° 22M) p- carboxylsenesulfonylazide. The ice bath was filtered, washed with acetonitrile, and the filtrate was concentrated to about 100 mL volume, then diluted with 800 mL ethyl acetate.

The organic solution was washed with aqueous sodium bicarbonate, brine and dried (MijSO ^. Evaporation of solvent afforded 55 g (90% yield) of ^l ouěen ^l n ^{y 4 f} Orme St. ^ETL e ^Zl utýc ^HK RYSTA] _u, m.p. ^{96 DEG} - ^{97 DEG} C.

IR (KBr) ^g 2120 and 1710 cm ^-1 . NMR (CDCl ^j) d ^e LT ^and 1.20 (3H, t, J = ^7. 0 H ^z b ^2, 85 (2H, q, J = 7.0 Hz), 5.40 (2H, s) 7.50 (2H, d, J = 8.0 Hz) and 8.15 (2H, d, J = 8.0 Hz).

Β. 1-p-nltrobenzyloxyk ^and rbonyl-L-diaza-2-tert-butyldimethylsilyloxy-2-butene (5) ^To ocHazenému ⁽⁰ ° ^C) solution of ^{54 g} (0.2M) of ^L ou en ^{No F} N ^Y A in ⁴⁰⁰ m ^{l methylcarbazole} enc ^hl or ^ld in added 41.4 g (° 4M) of TEA followed by 56 g (° 21M) t-butyldlmethylsilylchloridu in 30 ml of methylene chloride and during 40 minutes.

The solution was stirred for 120 minutes and then washed with ice water. The methylene chloride was dried (MgSO₄), filtered and evaporated in vacuo to give 68 g (89% yield) of compound jj, as ^TL in ^hydrous solid ^l let ^{K y} TT ⁵⁴ and ^{55 LC} (KBr) gamma ²⁰⁸⁰ and ^1,695 cm ¹

NMR of Compound 5 shows that Compound 5 is obtained as an E / Z mixture in the olefinic position.

113 9: 1 NMR (CDCl ₃ major isomer): delta 0.15 (6H, s), 0.90 (9H, s), 1.58 (3H, d, J = 7.0 Hz), 5.15 (2H, s), 7.30 (2H, d, J = 9.0 Hz) and 8.0 (2H, d, J = 9.0 Hz).

I OSi +

(

OSi 4 *

CO ₂ pNB

C. 4beta-1-Methyl-3-diazo-3-p-nitrobenzyloxycarbonyl-2-oxo-propyl) -3alpha-f- (R) -tert-butyldimethylsilyloxyethyl | -azetidin-2-one

To a suspension of 12.5 g (0.1 M) of anhydrous zinc chloride in 700 ml of methylene chloride was added 60.4 g (0.21 M) of compound 6, stirred at 23 ° C for 15 minutes and then cooled to 0 ° C. A solution of 106 g (0.27M) of compound (5) in 200 ml of methylene chloride was added dropwise to the reaction mixture over 90 minutes and then stirred for 120 minutes without cooling bath.

The reaction mixture was washed with aqueous sodium bicarbonate solution (4 x 150 mL), water, brine and dried over magnesium sulfate. Evaporation of the solvent gave a dark oil which was purified on a silica gel column.

Elution with 1: 9 ethyl acetate: methylene chloride gave 51.5 g (54%) of compound 2 ^as a white crystalline solid, mp 112-114 ° C. IR (KBr) gamma 2130, 1760 and 1720 cm ^1st

NMR at 360 MHz of compound ukazuje_ shows that compound Ί_ is a 2: 1 mixture of methyl isomers. NMR (CDC1 ₃₎ delta: 0.3-0.6 (6H, 2s), 0.82 (9H, 2s), 1.05-1.15 (6H, m), 2.68 (0.66 H , q, J = 6.6 and 2.0 Hz), 2.88 (0.34, q, J = 6.6 and 2.0 Hz), 3.57 (1H, m), 3.84 ( 1H, m), 4.09 (1H, m), 5.17 (2H), 5.84 (0.66 H, s), 5.95 (0.34H, s), 7.52 (2H, d, J = 8.5 Hz) and 8.23 (2H, d, J = 8.5 Hz).

OH b

CH ₃

NH ó

7>

CO ₂ pNB

114

D. 4beta- (1'Methyl-3-diazo-3-p-nitrobenzyloxycarbonyl-2-oxopropyl) -3alpha- [1. (R) -Hydroxyethyl] azetidin-2-one (8)

To a solution of Compound 7 (30 g, 59.5 ml) in 400 ml of thienyl was added 150 ml at 23 ° C.

1N hydrochloric acid and stirred for 18 hours. The reaction mixture was concentrated to about 200 mL and extracted with ethyl acetate (3 x 200 mL).

The combined ethyl acetate extracts were washed with water, aqueous sodium hydrogencarbonate solution and brine. Evaporation of the solvent dried over magnesium sulfate gave 22.3 g (96%) of Compound 2 as a white crystalline solid, m.p. Mp 147-148 ° C.

IR (KBr) ^g ama 3 ⁴ 0 ^{0 2} 35 ^{1 1} 7 ⁵ 0 cm ^-1 NMR ^EXAMPLE 360 MHz Compound ^Y it indicates that Compound # 8 was obtained as SMMS methyl at position 4 in the ratio 2: 1 . NMR (CMSO-d) δ: 1.07-1.10 (6H, m), 2.75 (0.66 H, q, J = 6.6 and 2.0 Hz), 2.85 (0, 34 H, J = 6.6 and 2.0 Hz), 3.55-3.90 (3H, m), 5.25 (2H, s), 7.70 (2H, d J = 9.0) Hz, 8.05 (0.66 H, s), 8.10 (0.34 H, s) and 8.27 (2H, d, J = 9.0 Hz).

Co ^ pNB

E. ppyitrobeyzylp6alpha [1- (R) phydropoyethyl)] p4pothylp3,7pdroo-1pazabicyclo [3.2.1 (T, heptane-2-carboxylate) (9)

A solution of 14.0 g (35.86 mmol) of compound (I) and 70 mg of rhodium (11) octanoate in ethyl acetate was refluxed for 20 minutes in a nitrogen atmosphere. The mixture was evaporated in vacuo to give compound 2 as a foam.

IC (CHCl3) ^g ama 3400 and 1750 cm ^-1 . ^EXAMPLE NMR 360MHz compound ^ n ^ y 2 shows ^that, with ^l ou en ^{i No.} 2 was obtained in the form of ISI may 4pmmthylu in position 2: 1st Nuclear Ovvrhauser effect is used to determine the 4-methyl configuration.

When it is irradiated with the H ₅ isomer, which is more, there is an approximately 7% increase in signal for 4-methyl protons, indicating the cis configuration and 4-methyl. Conversely, if there is less Η isomer of less, there will be no signal increase for 4-methyl, indicating a trans configuration and 4-pentyl for an isomer of less.

NMR (CDCl 3) of the isomer which is the more dylta: 1.24 (3H, d, J = 7.35 O, 1.40 (3H, d, J = 6.3 O, 2.40 (1H, m, 3, 24 (1H, q, J = 6.6 and 7.2 Hz), 3.67 (1H, q, J = 8.0 and 2.2 O, 4.18 (1H, m, 4.82 (1H) s), 5.24 (1H, d, J = 6.3Hz), 6.18 (1H, d, J = 6.3), 7.60 (1H, d, J = 8.5Hz) and 8.22 (1H, d, J = 8.5Hz).

NMR (CCCl-j) isomer of less delta: 1.0 (3H, d, J = 7m 35 Hzb 1.40 (3H, d,

J = 6.3 Hzb 2.83 (1H, m, 3.25 (1H, q, J = 6.6 and 1.50 Hz)), 4.14 (1H, q, J = 7.36 and 1, 50 HaO, 4.67 (1H, s), 5.24 (1H, d, J = 6.3 Hz, 6.18 (1H, d, J = 6.3 Hz) and 7.60 (1H, d, J = 8.5 Hz) and 8.22 (1H, d, J = 8.5 Hz).

115

₂ pNB

^{F. p} n ^{b of} TRL enн ^{yl -3 Dimethylpropy} phenoxymethyl ^{y phosphite yl} N - ⁶ and ^LF A- ^{£ l} - (R) -H ^ rox ^ th ^ J ^ beta-meth ^^ - oxo-lazabicykll (3,2,0) hept-2-ene-2-carboxylate (10) ^{to the} compound. ^{(O 0)} solution of ^{20.0 g} (5 ^5, 2 nunol) tetomez ^ roduHu Э_ ¹⁵ m ¹ 0 ^ HLU acetone was added 7.18 g (55 mmol)) dOisoprlpylethylαminu and then 14.85 g (55 mmol of diphenyl chlorophosphonate in 20 ml of acetonitrile over 5 minutes.

MMN ^^ m is ^I C 6 ° ^hook idnut p ^s 0 θ ^{0, C} o ^t om of a ^row of ^copper 6 ⁰ m ¹ et ^hyl αcet ^plus that rubs ice-cold 10% phosphoric acid and brine. The mixture was dried over magnesium sulphate and evaporated to give a crude oil, which was purified on a silica column.

Elution of the column with 10% ethyl acetate in methyleneiloride gave 3.7 g (11.5%) of phosphonate 10 as a white foam. IR (CHOlg) gamma 2400, 1790 and 1720 cm ^1st

NMR (CDCl3) δ: 1.20 (3H, d, J = 7.2 Hz), 1.38 (3H, d, J = 7.3 Hz), 3.35 (1H, l,

J = 6.7 and 2.0 Hz), 3.50 (1H, mu, 4.2-4.25 (2H, m)), 5.20 (1H, d, J = 10.5 Hz), 5 37 (1H, d)

J = 10.5 Hz), 7.1-7.4 (10H, m), 7.56 (1H, d, J = 9.0 Hz) and 8.10 (1H, d, J = 9.0 Hz).

Nuclear Overhauser effect was used to determine the 4-methyl configuration of compound 10. When irradiated with R -, there was no signal increase for 4-methyl- indicating the trans configuration - $ and 4-methyl.

CO ^ jNB

116

G. p-Nitrobenzyl-3- [pyridin-2-yl-methanthio] -6alpha-11- (R) -hydroxyethyl] -4-beta-methyl-7-oxo-1-azabicyclo [3.2.1] hept-2 -en-2-carboxylate (11)

To a cooled -15 ° C) solution of 1.2 g (2 mmol) of phosphonate 10 in 10 mL of acetonitrile was added 390 mg (3 mmol) of diisopropylethylamine followed by 370 mg (3 mmol) of 2-mercaptomethylpyridine under a nitrogen atmosphere.

The reaction mixture was stirred at -15 ° C for 60 minutes and then at 0 ° C for a further 60 minutes. The reaction mixture was diluted with ethyl acetate, washed with ice water, brine and dried (MgSO ₄ ).

Evaporation of the solvents in vacuo afforded a yellow oil which was purified on a silica column. Elution of the column with 20% ethyl acetate in methylene chloride afforded 375 mg (40% yield) of compound 11 as a white amorphous foam.

IC (KBr) gamma 3400, 1775 and 1710 cm ^-1 . NMR (CDCl ₃ ) δ: 2.14 (3H, d, J = 6.7 Hz), 2.19 (3H, d, J = 6.7 Hz), 4.14 (1H, q, J = 6) , 2 and 2.0 Hz), 3.40 (1H, m), 4.0 (1H, d, J = 7.6 Hz), 4.18 (1H, q, J = 6.7 Hz) and 2.0 Hz), 4.25 (1H, m), 5.25 (1H, d, J = 11.3 Hz), 5.40 (1H, d, J = 11.3 Hz), 7, 15-8.2 (4 H, m).

OH CH ₃

H. 3- [2- (N-methylpyridinium) methanthio] -6alpha- [1- (R) -hydroxyethyl] -4-beta-methyl-7-oxo-1-azabicyclo [3.2.0] hept-2 -en-2-carboxylate (12)

To a solution of compound 11 (1.0 g, 2 mmol) in methylene chloride (10 mL) was added methyl trifluoromethanesulfonate (450 mg, 3.3 mmol) and stirred at 23 ° C for 90 min. Evaporation of the methylene chloride in vacuo gave the quaternized pyridine as a foam which was hydrogenated immediately without further purification.

The crude pyridinium salt was dissolved in TMF-ether buffer pH 7 (1 - 1, 100 ml each), then 600 mg of 10% palladium on carbon was added. The mixture is hydrogenated for 45 minutes at 20 psi on a shaker. The mixture was filtered through celite and the catalyst was washed with water (2 x 10 mL).

The combined filtrate and washings were extracted with ether (2 x 100 mL) and lyophilized to give a yellow powder which was purified on a reverse phase C 10 g Bondapak column, eluting with 5% acetonitrile in water at 55 kPa.

Each 15 ml fraction was subjected to high pressure liquid chromatography and the ultraviolet absorption fractions at lambda _max 300 nm were combined and lyophilized to give 50 g (11% yield) of the title compound as a pale yellow powder.

117

Μ. "1 ^IR (KBr) GA 3400 and ^{m ',} 1" 750 ^{and 1590} cm. MBDA ^ ^ (H ₂ O) 292 nm (EPS-one ^{7 08} 1).

NMR (D ₂₀ ) δ: 1.13 (3H d, J = 6.5 Hz), 1.23 (3H, d, J = 6.5 Hz), 3.18 (1H, m), 3.45 ( 1H q, J = 6.0 and 2.1 Hz), 4.0-4.4 (4H, m), 4.65 (3H, s), 7.79 (2H, m)

8.30 (1H, m) and 8.60 (1H,

(m)

he said

Execution 3- ^£ 2- (l, 4- ^di me ^t h ^{ylpyridine i} s ⁱ n ⁱ m ^ methanthwj alpha ^{- [1} (R) ^{- j} h ^yd roxyethyl -4beta meth ^{yl-1-7-oxo-l-} azabicylo (3.2.0) hept-2-ene-2-carboxylic ^yl methacrylate

2) CF 3 SO 3 CH 3

3) Pd / H ₂

3 ^{- (2 -} (d ^^ ^ et ^ LP ^ r ^ CHR ^ umúmethanthio] ^-6 alpha ^- [l ^{- (R)} -h ^{y y} Hydroxy eth ^{yl] -4b} ETA ^- meth ^y l- oxo ⁷ -l-azabicyclo (3,2,0) hept-2-ene-carboxylate ^yl methacrylate (13)

This compound was obtained as a yellow powder in 17% yield from the compound in the same manner as described in Example 27. IR gamma 3400, 1755 and 1600 cm

Ю

UV

UV λ _max (λ 0) 300 nm (epsilon 7,600).

NMR (1 H 2 O) δ 1.20 (3H, d, J = 6.7 Hz), 1.28 (3H, d, J = 6.7 Hz), 2.60 (3H, d, J = = -6.7 Hz), 1.28 (3H, d, J = 6.7 Hz), 2.60 (3H, s,) 3.4-3.5 (2H, m), 4.2-4 4 (4H, m), 4.52 (3H, s), 7.82 (1H, t, J = 6.5 and 4.2 Hz), 8.32 (1H, d, J = 6.5 Hz) and 8.60 (1H, d, J = 4.2 Hz).

Example 29 Preparation 3 ^{- [4 - (l-methylpyridin} ^^ wmmetihantihiq] ^- 6a ^{LF -} [1- ^(R) ^ Hydroxy ^ tihylj ^- 4beta ^- meth ^{yl-7 -}

oxo-1-azabicyclo (3.2.0) hept-2-ene-2-carboxylate

U HS

2) CF ₃ SO ₃ CH ₃

3) Pd / H2

118

3- (4- (1-methylpyridinium methanthio) 6 alpha - [1- (R) -hydroxyethyl] -4-beta-methyl-7-oxo-1-azabicyclo (3.2.0) hept-2-ene-2-carboxylate (14)

This compound was obtained as a yellow powder in 15% yield from compound 10 in the same manner as described in Example 27.

IR (KBr) gamma 3410, 1750 and 1650 cm ^-1 . UV λ _max (H ₂ O) 293 nm (epsilon 7,295).

NMR (D ₂ O) δ: 1.15 (3H, d, J = 6.5 Hz), 1.20 (3H, d, J = 6.5 Hz), 3.20 (1H, m), 3 45 (1H, q, J = 6.0 and 2.0 Hz), 4.11 (1H, q, J = 8.0 and 2.0 Hz), 4.20 (1H, m) and 4, 35 (3H, s);

7.95 (2H, d, J = 5.2 Hz) and 8.72 (2H, d, J = 5.2 Hz).

Example 30

Preparation of 3- [α- (1-methylpyridinium methanthiq) -6alpha. Q- (R) -hydroxyethyl] -4-beta-methyl-7-oxo-1-azabicyclo (3.2.0) hept-2-ene-2-carboxylate

«'-Q

2) CF ₃ SO ₃ CH ₃ --------->

3) Pd / H ₂

3- (α- (1-methylpyridinium) methanthioyl-6α-α- (R) -hydroxyethyl) -4abeta-methyl-7-oxo-1-azabicyclo (3,2,0) hept-2-ene-2-carboxylate (15)

This compound was obtained as a yellow powder in 27% yield from compound 60 in the same manner as described in Example 27.

IR (KBr) gamma 3420, 1750 and 1610 cm UV λ _max (H ₂ O) 295 nm (epsilon 8 750). NMR (D ₂ O) δ: 1.10 (3H, d, J = 6.9 Hz), 1.25 (3H, d, J = 6.9 Hz), 1.27 (1H, m), 1 43 (1H, q, J = 6.2 and 1.8 Hz), 4.1-4.35 (4H, m), 4.39 (3H, s), 8.0 (1H, ie, J = 8.5 and 6.2 Hz), 8.45 (1H, d, J = 8.5 Hz), 8.70 (1H, d, J = 6.2 Hz) and 8.82 (1H, s) ).

119

Annlyza: calcd for C ₁₇ H ,, ^ θΝ δ. 2 2.1 H ₂ 0: C, 51.90, H, 6.36, N, 7.12; found: C 51.92, H 5.71, N 6.88 · Example 31

Preparation

3- ^[3- (1, R₁ 'dimethyl ^ y) at ^d L ^p L Hiimúm}! ^? ^^^ and h.Ío] -6alfa- ^[1- (R) -hydroxyethyl] -4beta-methyl- 7-oxo-1-azabicyclo (3,2,0) hept-2-ene-2-carboxylate

</.

3) СТзБОзСН ₃

3) Pd / H ₂

''and

³ ··· [з ~ (^] ,²-dimt^thy^pyrri ^ rdumúnitthariyhi ^^-6al^Fand^{- [1-} (^RIbetamia^thyl^-7-ooo -1 -a2sbic^yklo (3,2, 0) Уерк-2-on-Ž-carbo ^ y L (16)

This compound is obtained as a yellow powder in 14¾ yield from compound 1Q as described in Example 27,

Tc (КВг) ^{3 41 g} AMA °, ¹⁷⁵⁰ NMR (B ₂ O) delta: 1.25 (311, d, J (1H, m), 3.50

J = 8.5 and 6.3 (1H, q, J = 6, -2 and

H, 8.40 (1H, d, and ¹⁶⁰⁰ cm ³ UV lambddd ma).

= 6.5 H), 1.30 (3H, d, J 1.8 Hz). 4.2-4.4 (4 H, m),

J = 8.5 Hz) and 8.72 (1H, d _at (1 ^H ₂ )) 29 ⁶ nm (epsilon 8 500).

= 6.5 Hz), 2.95 (3H, s), 3.40

4.35 (3H, s), 7.82 (1H, t, J = 6.3 Hz) ·

Example

Preparation of [3- (2? ^ • drniiet.bu / i · L 2 - lt.LiazolÁum) -mithanthl-oj --6al ^F N- [l- ^{(R) '-} H ^y ílrox ^y eth ^y l ] - ^4b eta ^ -m · Ithylo ^ - 7 · ·· oxo-1 - azabicyclo · '3, 2.0) 2-- hcpt- tn-2 - carboxylate

Pl / ll?

120

[3- (2,4-dimethyl-1,2,4-triazolium) methanthio] -6alpha- [1 (R) -hydroxyethyl] -4-beta-methyl-7-oxo-1-azabicyclo (3.2.0) Hept-2-ene-2-carboxylate (17)

This compound was obtained as a yellow powder in 9% yield from compound 10 in the same manner as described in Example 27.

IR (KBr) gamma 3420, 1756 and 1725 cm UV λ _max (H ₂ O) 291 nm (epsilon 7 850),

NMR (D ₂ O) δ: 1.15 (3H, d, J = 6.3 Hz), 1.22 (3H, d, J = 6.3 Hz), 3.35 (1H, m), 3 48 (1H, (1H, q, J = 6.0 and 1.8 Hz), 3.90 (3H, s), 4.05 (3H, s), 4.2-4.4 (4H, m), 8.80 (1 H, s).

Example 33

Preparation of 3- {2- (1,3-dimethylimidazolim) methanthioj-6alpha- [1- (R) -hydroxyethyl] -4beta

methyl 7-oxo-1-azabicyclo (3.2.0) hept-2-ene-2-carboxylate; ^H 3

OH 1) 2) hs ^ <N ^CF 3 ^SO 3 ^CH 3 0 ^ 1 CO ₂ pNB IQ 3) Pd ^{/ H} ₂

3- {2- (1,3-Dimethylimidazolium methanthio) -6alpha- [1- (R) -hydroxyethyl] -4-beta-methyl-7-oxo-1-azabicyclo (3,2,0) -hept-2-en-2 carboxylate (18)

This compound was obtained as a yellow powder in 32% yield from compound 60 in the same manner as described in Example 27.

IR (KBr) gamma 3400, 1758 and 1600 cm UV λmax (H ₂ O) 294 ^nm (epsilon 7,194).

NMR (D ₂ O) δ: 1.10 (311, d, J = 6.3 Hz), 1.25 (3H, d, J = 6.3 Hz), 3.30 (1H, m), 3 42 (1H, 1, J = 6.0 and 2.2 Hz), 3.85 (6H, s), 4.2-4.6 (4H, m) and 7.40 (2H, s).

121

Example 34

The following carbapenem products are prepared according to the general procedures of the intermediate of formula 1 to ₂ pnb.

Example

No. A

34a-OH ₂ 34b -CH ₂ CH ₂ 34c 'C ^H 2

4d -ch ₂ ;

ch ₃ \ e

34e -CH ₂ CH ₂ -

34f

-ch ₂ -

122 continued

Example

C.

tables

34g ^CH 2 “

CH ₃

34h “ ^CH 2”

34i ^CH 2 “

CH ₃

34j

-CH ₂ -

34k

-CH ₂ -

341

4m

- ^CH ₂ '

34n

CH ₃

34o

(mixture of 3 possible isomers)

CH ₃

123 continued table

Example

No. A

34p — N и Θ _S xM-CH ₃

34q “CH ₂ -

34r

TO

34s ^CH 2 “

I®

Y _N —N

AND

CH ₃

34t

-ch ₂ -

Example 35

Following the general procedures of Examples 1 to 33, the following carbapenem products were prepared from the intermediate of formula

Example #

35a

-C ^CH ₂ 35b

-ch ₂ ch ₂ 124

35c ^-c V 35d ^{-C 2-} 35e ^-C V

^ ° ⁿ ®CH ₂ CH ₂ CH ₃

CH ₃ ® ^c '4 ~ l ^ch '

35f - ^CH 2- - /] N- N- ³ 1 CH ₃ 3 5g ? ^H3 -CH- O- ^c j 35h CV X-) CH ₃ ^Z 35i - ^CH 2- ^{, Shrinkage of 3} possible isomers) CH ₃

35j

—N

II Θ s' ⁿ -ch ₃

125

continued table

Example

č.A

35k ~ CH2351 ~

¹ A ch ₂ -coo

35m

-ch ₂ -

35n -CH ₂ СНз

N-N-CH, -4 II

CH ₃

Example 36

When the ketone intermediate is replaced by an equimolar amount of the corresponding 1-alpha-methylmethyl intermediate, the above carbapenem product is obtained.

Claims (7)

SUBJECT OF THE INVENTION 1 8 2 ' 1) reacting rea ^g OVA ^t between ^P ro ^d u ^k t general ^ o formula Ы1 wherein ^R, R and R · have ^the meanings given in point 1 and R ¹⁵ is a hydrogen atom in an inert organic solvent with a reagent capable of introducing a normal the leaving group L at position 2 of the formula III intermediate to give neziprodu ^kt in general ^ o formula ^{I, to} de ^r \ ^r8 and ^R2 approached ^the meanings Uve ^d said preceding ^b of ^{dE 1} and ^R18 is hydrogen, and L is a conventional Outgoing group 1 8 2 * 1) reacting an intermediate of formula III (III) wherein R ^1, R 8 and R @ ¹ are as defined above and 1-4 nitrogen, oxygen or sulfur atoms and attached to A through a ring carbon atom and having a ring nitrogen atom which is quaternized by R and their pharmaceutically acceptable salts, characterized in that: A process for the preparation of carbapenem derivatives of the general formula I wherein g R is hydrogen and R ^{1 is R a} Y b not that ^when n ^y SKUs for rnuj ^{H I} C ^I by treatment with ^, ^^ Subs alkylated ^or nesub ^{STI's o} ý ^an alkyl, alkenyl, and alkynyl having up to 10 carbon atoms, cycloalkylalkyl having (3-6C) cycloalkyl ring and (1-6C) alkyl, phenyl, 126 arylalkyl, arylalkenyl and arylalkinyl wherein the aryl moiety is phenyl and the aliphatic moiety has up to 6 carbon atoms, heteroaryl, heteroarylalkyl, heterocyclyl and heterocyclylalkyl, wherein heteroaryl is a mono-, di- or polycyclic aromatic heterocyclic radical having 5 or 6 members in individual rings and heterocyclyl is a mono-, di- or polycyclic saturated or unsaturated non-aromatic heterocyclic radical having 5 or 6 members in individual rings and wherein the heteroatom or heteroatoms in the above-mentioned moieties are selected from the group consisting of 1 to 4 oxygen, nitrogen or sulfur atoms and alkyl moieties attached to said heterocyclic moieties have 1 to 6 carbon atoms, wherein the substituent or substituents of the above radicals are independently selected from the group consisting of alkyl of 1 to 6 carbon atoms, optionally substituted with amino, halogen, hydroxyl or carboxyl, halogen -0Y II 3 4 -OCNR R O Ί 34 4 -CNR ^J R 2 'optionally replacing anion X' with another anion and optionally removing the carboxyl protecting group R to form the desired unprotected compound of formula I or pharmaceutically acceptable salts thereof. 2) then was allowed to intermediate of formula IV to react in an inert organic Inhalants and in the presence of a base with a mercaptan of the formula specified in point 1, to give 18 2 'of the intermediate of formula II wherein R, R, A, R, and R ¹⁵ has the meaning given in this point, 2 ', the anion X is replaced by another anion and optionally the R 1 carboxyl group is removed to give the desired unprotected compound of formula (I), or a pharmaceutically acceptable salt thereof. 2) further reacting the intermediate of formula IV wherein R ^1, R ^8, L and R are as defined in ^b of ^DE 1 and ^{R 2 8} is Aton in ^thanks UV iner ^t n ^e n organtokém solvents: ^{eliminated in} EAV pMtdnnosti Base, nerkaptanem general formula specified in point 1, to give the intermediate of formula II ^to de ^{R2, R8,} a, R ² and Z 'have the meanings given in point 1, and ^OR28 represents a ^{thank u} 2. ^A method of making a ^{p yk} arba enenových obecnéto derivative ^of the formula I according to item ² wherein ^{R, R 2, R 8, R.} and ^the meanings and NAJ Uve ^{d b} of said preceding ^{DE 1} and R ²⁸ is a hydrogen water ^ u, ^wherein labeled ^by 2 ', the carboxyl protecting group R is removed to give the desired unprotected compound of formula I or pharmaceutically acceptable salts thereof. 2) The intermediate of formula IV is reacted in an inert organic solvent and in the presence of a base with a mercaptan of the formula wherein A is as defined above and represents a substituted or unsubstituted mono- or polycyclic Ы- aromatic heterocyclic radical containing at least one nitrogen atom ^{in the} ring and a total of 1 to 4 nitrogen, oxygen or sulfur atoms and containing a quaternizable ring nitrogen atom attached to A via a carbon atom to form an intermediate of formula (II) wherein they are as defined above, 2, R 'is a conventional carboxyl protecting group which is easily cleavable and selected from the group consisting of benzhydryl, allyl, p-nitrobenzyl, 2-naphthylmethyl, benzyl, trichloroethyl, trialkylsilyl wherein alkyls have 1 to 4 carbon atoms, phenacyl, p-methoxybenzyl, acetonyl, o -nitrobenzyl, 4-pyridylmethyl, C 1 -C 6 alkyl, pivaloyloxymethyl, acetoxymethyl, phthalidyl, indanyl, methoxymethyl and preferably p-nitrobenzyl, in an inert organic solvent with a reagent capable of introducing a common leaving group L to the 2-position of the intermediate of formula III to give intermediate IV 130 (IV ^) where R ^1, R ^8, R ¹⁵ and R ² are as defined above Uve ^d Eny ^{EA L BE} is Zn ^and ODS upuj ^{t H} C ^H skuptoa, 3) The intermediate of formula II is reacted in an inert organic with an alkylating agent of the formula R $ - X _'where а X' have the meanings indicated 18 2 'to give the intermediate of formula wherein R, R, A, R, f \ _{c 15 Dec} X and X are as defined in point 1 and R is as defined in point 1 and solvent is in point 1 3. A process karbapenenových derivatives of general formula I according to claim 1, where R ', R ^2, R ⁸ and R ⁸ and A approached ^the meanings defined in ^b of ^DE-A and ^R32 is substituted and the net ^b b ^b itucvaný and ^lkyl, alkenyl and alkynyl having up to 10 carbon atoms, cycloalkyl, cyкOcal ^к ylalk ^к yl alkylcycloalkyl having 3-6 carbon atoms in ^CY 0cal ^to ylovéш ring and 1-6 carbon atoms in the alkyl moieties, tpirccyklcalkyO having 3-6 carbon atoms, phenyl, , arylalkyl, arylalkenyl and arylalkinyl wherein the aryl moiety is phenyl and the aliphatic moiety has 1 to 6 carbon atoms, heteroaryl, heteroarylalkyl, heterocyclyl and heterocyclylalkyl, wherein the heteroaryl is a mono-, di- or polycyclic aromatic heterocyclic radical having 5 or 6 members in each and heterocyclyl is a mono-, di- or poly-saturated or unsaturated non-aromatic heterocyclic radical having 5 or 6 members in each ring and wherein the heteroatom or heteroatones in the above-mentioned heterocyclic moieties are selected from and the alkyl moieties attached to these heterocyclic moieties have from 1 to 6 carbon atoms. carbon atoms wherein the substituent (s) of the above radicals are selected from the group consisting of amino, mono-, di- and trialkylamino, hydroxyl, alkoxy, mercapto, alkylthio, phenylthictocupine, sulfamoyl, amidino, guanidino, nitro, chloro, bromo, bromo , cyano and carboxyl and wherein the alkyl moieties in the above substituents have 1 to 6 carbon atoms,. characterized by: 18 2 '/ z. (1) reacting an intermediate of formula (III) wherein R, R and R are as defined in (1) and R ( ¹⁾ is as defined in an inert organic solvent with a reagent capable of introducing a common leaving group L to the 2-position of the nonproduct ^to de R ^3, R ⁸ and ^{R 2} are as defined in ^{b of} from 1 · ²⁸ m R ^and importance ^d Uve said preceding -tomto b ^ E 132 3) The intermediate of formula II is reacted in inertnín organic with an alkylating ^ rea ^ m of the formula R ^8, X ', ^w e ^{R 8} and X' have ^the meanings Uve ^d Eny 18 2 for MMN ^also with remembered again ^P ro ^d u ^{k eh} of the general formula ^{I 'kd} e ^{R, R, a,} R ^2, and R ¹⁵ is optionally a solvent in point 1 of hydrogen and protecting Aton 3) reacting the intermediate compound of formula II with an alkylating agent of formula II in an inert organic solvent - X 'where c _ I R is as defined above and X 'is a conventional leaving group, such that R is quaternized with a ring nitrogen atom in the intermediate of formula II to form a compound of formula I xO d) 131 is as defined above, and optionally replaces anion X 'with another anion and optionally 3 4 -CONR ³ R ⁴ -CN, or phenyl optionally substituted with 1 to 3 fluorine, chlorine, bromine, alkyl of 1-6 ΊΊ4 3 3 34349 at ^om y carbons -OR "NR ³ R \ -80 C ^% ^<O, R ³ or -CONR ³ R 1. where R ³ ,. R ⁴ and R ⁹ in the substituents R · have the meanings indicated above, no ^b of R ⁵ m ^already be ^connected, onnection ^E on the other side of the ring so as to form a fused heterocyclic or hetercarcnatický ring which may contain further heteroatoms selected from oxygen, nitrogen and sulfur groups, R ¹⁵ is hydrogen, substituted and unsubstituted alkyl, alkenyl and C 1 -C 10 alkyl, cycloalkyl, cycloalkyl and C 3 -C 6 alkylcycloalkyl having 1 to 6 carbon atoms in the alkyl moieties, spirocycloalkyl. 129 having 3 to 6 carbon atoms, phenyl, arylalkyl, arylalkenyl and arylalkinyl wherein the aryl moiety is phenyl and the aliphatic moiety has up to 6 carbon atoms, heteroaryl, heteroarylalkyl, heterocyclyl and heterocyclylalkyl, wherein heteroaryl is a mono-, di- or polycyclic aromatic heterocyclic the ring having 5 or 6 members in each ring and the heterocyclyl is a mono-, di- or polycyclic saturated or unsaturated non-aromatic heterocyclic ring having 5 or 6 members in each ring and wherein the heteroatom or heteroatoms in the above-mentioned heterocyclic moieties are selected from the group consisting of 1 to The 4 oxygen, nitrogen or sulfur atoms, and the alkyl moieties attached to these heterocyclic moieties, have 1 to 6 carbon atoms, wherein the substituent (s) of the above moieties are selected from the group consisting of amino, mono-, di- and trialkylamino, hydroxyl, alkoxy, mercapto, alkylthio, phenylthio, sulfamoyl, amidinos upine, guanidino, nitro, chloro, bromo, fluoro, cyano and carboxyl, and wherein the alkyl moieties in the above substituents have 1 to 6 carbon atoms, A is a straight or branched alkylene having 1 to 6 carbon atoms, R is a hydrogen atom, an anionic charge, or a conventional carboxyl protecting group which is easily cleavable, selected from the group consisting of benzhydryl, allyl, p-nitrobenzyl, 2-naphthylmethyl, benzyl, trichloroethyl, trialkylsilyl wherein alkyls have 1 to 4 carbon atoms, phenacyl, p- methoxybenzyl, acetonyl, o-nitrobenzyl, 4-pyridylmethyl, alkyl of 1 to 6 carbon atoms, pivaloyloxymethyl, acetoxymethyl, phthalidyl, indanyl, methoxymethyl and preferably p-nitrobenzyl, wherein when R is hydrogen or a protecting group, an ion is also present and the group represents a substituted or unsubstituted mono-, bi- or polycyclic aromatic heterocyclic radical containing at least one ring nitrogen atom and overall 3 ' ¹ 9 -NR ³ SR ⁹ clay O -SR S ³ -SO ³ -R ³ -CO ² R ³ Wherein, in the above substituents, R and R are independently selected from the group consisting of hydrogen, alkenyl and alkynyl of up to 10 carbon atoms, cycloalkyl, eucloalkulalkyl, and alkyl of 3 to 6 atoms in the eucloalkyl ring and s; C 1 -C 6 alkyl moieties, phenyl, arylalkyl, arylalkonyl and arylalkinyl wherein the aryl moiety is phenyl and the aliphatic moiety has 1 to 6 carbon atoms and helene ^ 1, heteroarylalkyl wherein the heteroaryl is a mono-, di- or semi-nuclic aromatic ieterocuccinic a ring having 5 or 6 members in single rings and a helical ring is a mono-, di- or polyeucyl saturated or unsaturated non-aromatic 5 or 6 membered ring hetero ring and wherein the heteroatom or heteroatoms are selected from the group consisting of 1 to 4 atoms oxygen, nitrogen or sulfur, and the alkyl 3 4 moieties associated with the ieterocuclic moieties have 1 to 6 carbon atoms or R and R together with the nitrogen atom to which they are attached at least one is attached may form _zz 9 five or six membered nitrogen-containing heterleuklický ring, R is uvede3 18 Ny for ^{R to} ROM ^E is water ^Cart with Neto ^R and ^R in ⁱ rlmαd ^u constitutes an AU ^ Hen with ^2-10 C atoms or (C 2 -C 10) alkylidene substituted with hydroxy; R ⁵ is substituted or unsubstituted alkyl, alkenyl and alkynyl of up to 10-carbon atoms, cycloalkyl and (C 1-6 -alkyl) of (C 1 -C 6) -cycloalkyl ring and C 1 -C 6 -alkyl, phenyl, arylalkyl, arylalkenyl and arylalkonyl wherein the aryl moiety is phenyl and the aliphatic moiety has up to 6 carbon atoms, heteroaryl, heteroarylalkyl, helenyl, and heteroaryl, wherein heteroaryl is a mono-, di- or pllycucyl aromatic heterocyclic radical having 5 or 6 members in individual rings and the heterocyclyl is a mono-, di- or semi-carbon saturated or unsaturated non-aromatic heterocyclic radical having 5 or 6 members in individual rings and wherein the heteroatom or heteroatoms in the above said alkyl moieties are selected from the group consisting of 1 to 4 oxygen, nitrogen or sulfur atoms, and the alkyl moieties associated with said heterocyclic moieties have 1 to 6 carbon atoms, wherein the aforementioned R radicals are the case; substituted with 1 to 3 substituents independently selected from the group consisting of alkyl of 1 to 6 carbon atoms, optionally substituted with amino, fluorine, chlorine, carboxyl, hydroxyl or carbannyl, fluorine, chlorine or bromine; OR ³ -OOO ² R ³ -ocor ³ 128 -OCONR ³ R ⁴ II -OS-R -0X0 R ³ CONR · 34 _NR ³ CO2R 33 4 -NR ³ CONR ³ R O 3 4 -NR ³ CO2R ⁴ - ^NO 2 3 P 9 -NR ° S-R 11 O -NR 3 4 -NHCNR R ' ³ ' ⁴ ' ^CO 2 ^r3 -0P (0) (OR ³ ) (OR ⁴ ) = 0 ¹¹ 3 -OCR ii -SR sr ' -CN 127 - ^N 3 -OSO ₃ R ³ O H 9 -OS-R II O O 3 4 -SO ₂ NR R O 4. The process according to claim 1, wherein the quaternization step is carried out 2 'after the removal of the carboxyl protecting group R. 4 O = ^NR4 l3 5. The process of claim 2, wherein the corresponding starting compounds of step 2 are reacted together to give 3- (N-methylpyridin-2-yl-methanthio) -6alpha- [1- (R) -hydroxyethyl] - 7-oxo-1-azabicyclo (3.2.0) hept-2-ene-2-carboxylate. 6. The process of claim 3, wherein the corresponding starting compounds of point 3 are reacted to give 3- (N-methylpyridin-2-yl-methanthio) -4alpha-methyl-6alpha- [1- (R)]. -hydroxyethyl] -7-oxo-1-azabicyclo (3.2.0) hept-2-ene-2-carboxylate. 7. The process of clause 3, which comprises reacting the corresponding starting compounds of clause 3 to form 3- (N-methylpyridin-2-yl-methanthio) -4beta-methyl- 6-alpha - [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo (3.2.0) hept-2-ene-2-carboxylate.