CS271490B2 - Method of new acylderivatives production - Google Patents

Abstract

3-Acylated-methyl-cephalosporin derivs. of formula (I), their easily hydrolysable ester, pharmaceutically acceptable salts, and their hydrates are new: X = OCO, SCO, NHCO or OCONH; R1 = acyl; R2 = H or lower alkoxy; R3 = a gp. (II)-(IV); R4, R4' and R4'' = H or COR2; R2 = opt. branched lower alkyl; A and B = halo; a, b, x, y and z = zero (corresp. to H at that position) or 1, at least 2 of x, y and z are 1; m = 0-8. Also new are intermediates (XVIII) of similar formula but with R1 replaced by H and the 4-COOH gp. opt. protected. Pref. m = 0 or 1; R4, R4' or R4'' = H or MeCO; A and B = chloro; R = H.

Description

(57) Compounds of formula I wherein X is -000-, -SCO-, -NHCO- or -OCOMH-, R 1 is acyl of up to 21 carbon atoms, is hydrogen or C 1-8 -alkoxy and R 3 is of formula (a), (b) or (c), wherein R 4, R 4 * and R 4 'are each independently hydrogen or a -COR 200 group wherein R 200 is straight or branched C 1-6 -alkyl, A and V are halogen, and, b, x, у and z are each independently zero, and in this case, at the corresponding position _{w of the} ring, there is hydrogen, or the number 1, with at least two of the indexes x, у and z being l The number from 1 to 8, as well as their pharmaceutically usable salts and hydrates of the compounds of formula I, or salts thereof, are prepared by cleavage of the protecting group R from the ester of formula XIX. The compounds produced have antibacterial activity and can be used as medicaments.

i 1

71490 В2 ej

CO o

Italy (11) (13) B2 (51) Int. Cl. ⁵ c 07 A 07 D 501/26 D 501/36 К 31/525

* 2 ^H _

COOR

CS 271 490 B2

The present invention relates to a process for the preparation of novel acyl derivatives having valuable pharmacological properties.

New acyl derivatives of the formula I have been found

(AND)

in which

X is -OCO-, -SCO-, -NHCO- or -OCONH- *

Rj represents an acyl group * which is used to acylate β-lactam antibiotics and which contains up to 21 carbon atoms * for example a group of formulas

^R 101 - ^CH 2 ^C ° -

R ₁₃₀ -O-N = C-CO ^{1 R} 101 wherein ^R 101 ^{denotes a} 5-membered heterocyclic ring containing 1 or 2 nitrogen atoms and / or sulfur atoms * 'and which is optionally substituted with an amino group * ^R 120 ^denotes an alkyl group having 1 to 2 8 carbon atoms and ^R @ 130 ^marking alkyl group having 1-8 carbon atoms, * optionally substituted with carboxy including a salt form carboxyl, carbamoyl or alkoxycarbonyl with 2-9 carbon atoms *

R ₂ represents hydrogen or alkoxy having 1-8 carbon atoms and

R ₃ represents a group of the general formula

CS 271 490 92

(or ₄ -) ₂

‘®Г>,

Wherein R @ 4 * R @ 4 * ^and R @ 2 are each independently hydrogen or -or ₂₀₀ , wherein:

Rgoo ^ ^represents straight-chain or branched alkyl having 1 to 8 carbon atoms, А а В represents a halogen atom;

and; b; x; у and z are each independently zero (in these circumstances a hydrogen atom is present at the appropriate position of the ring) or the number 1 β by at least two of the indices y; x and z are each 1, and m is zero or an integer from 1 to 8, and the pharmaceutically usable salts of these compounds and the hydrates of the compounds of formula I or their salts;

The term alkyl *; as used herein refers to straight chain or branched saturated hydrocarbon groups; which contain 1 to 8 carbon atoms; and preferably contain 1 to 4 carbon atoms. Deco examples include methyl, ethyl; n-propyl; i-propyl; tert-butyl group, etc. ·

The term alkoxy *; as used herein refers to a straight or branched chain hydrocarbon-oxy group; wherein the alkyl moiety is represented by an alkyl group having 1 to 8 carbon atoms, as defined above; Oako examples include methoxy; ethoxy; n-propoxy, etc. ·

The term halogen *, as used herein, refers to all four forms, ie, · chlorine, bromine, iodine and fluorine, unless otherwise indicated ·

A 5-membered heterocyclic ring containing 1 or 2 nitrogen and / or sulfur atoms means, for example, the following groups: pyrazolyl, pyrrolidinyl; an imidazolyl group, a thlazolyl group; thisnyl group etc ·

As a substituent, for example, a 2-amino-4-thiazolyl group may be present. As acyl groups of up to 21 carbon atoms are meant groups which may be used to acylate antibiotics (1-lactam type; including 6 * aminopenicillanic acid; as well as 7-aminocephalosporanic acids and their derivatives: cf., for example, Cephalosporins and Penicillins, edited by Flynn, Academic Prese (1972), Belgian Patent 866 038, published 17 · 10 · 1978, Belgian Patent No. 867 994 , U.S. Patent No. 11,112,478, U.S. Patent No. 4,152,432, U.S. Patent No. 4,152,432, U.S. Patent No. 4,152,432, U.S. Patent No. 4,171,478, U.S. Patent No. 3,971,778, issued July 27, 1976, and U.S. Patent No. 4,173,199; 23 · 10, 1979 · In these publications, ee lists various acyl groups which ee use in the present case · The following acyl groups further illustrate the term acyl group; dným way limit · H. Examples of acyl groups having up to 21 carbon atoms include:

(a) a carbocyclic aromatic group of the formula

(b) heteroaromatic groups of the general formula

RjqjL “CHg CO”

CS 271 490 B2 wherein ^R 101 is ^an optionally substituted 5-membered hernocyclic ring containing 1 or 2 nitrogen and / or sulfur atoms; examples of hotsrocyclic rings include thienyl, pyrrolyl, pyridinyl, pyrazinyl, thiazolyl and pyrimidinyl;

The amino group may be present, for example, as a 2-amino-4-thiazolyl group.

Preferred heteroaromatic acyl groups include acyl groups of the above formulas; wherein R _lol represents 2-amino-4-thiazolyl, ¹ 4-aminopyridin-2-yl; 2-thienyl or

4-pyridinyl · (c) // (4-aubatituated 2,3,3-dioxo-1-piperazinyl) caronyl / amino / arylacetyl group of formula

wherein ^R 120 ^is 2-C 1 -C 8 -alkyl (d) (substituted oxyimino) arylacetyl groups of the formula

R 23Q - ·· O · C * N ^R 101 CO ^R 101 wherein the above meanings and ^R 130 represents an alkyl ekupinu e 1 to 8 carbon atoms which is optionally substituted with carboxy (karboxyekupiny including in salt form), carbamoyl

or an alkoxycarbonyl group having from 2 to 9 carbon atoms As examples of the general group formulas ^R 1.CO - 0 - N - c i - what - F ^ř 101 UV‘Ul H А «» 1 4 »<iu I i <* t <llM4 | tl4ty«

2- (2-Aminothiazol-4-yl) -2-isopropoxyiminoacetyl, N - (2-aminothiazol-4-yl) -2-methoxy-aminomethyl,

2-t-hionyl-2-mothoxylminoaoyl,

2- (2-aminothiazol-4-yl) -2- (carboxymethoxyimino) asyl, 2- (2-aminothlazol * 4 * yl) -2- (carboxy-propoxyimino) acetyl, etc. ·

A preferred group of compounds are those of formula II

CS 271 490 B2

COOH

CH ₂ XR ₃ (II) wherein

X; R 8 and R ₃ are as defined above; and

Rg1 represents a hydrogen atom, an alkyl group having 1 to β carbon atoms or a group of the general formula

- - COOH * 23 where

Rg & 23 represents a hydrogen atom or an alkyl group having 1 to 8 carbon atoms or in common and a carbon atom; to which they are bound; form a carbocyclic ring having from 3 to 7 carbon atoms. More preferably, the compounds of formula (II) wherein R 8 is hydrogen and R 8 is methyl or formula

COOH in which ^R 22 ^{and R} 23 represent a hydrogen atom or a methyl group ·

A further preferred group of compounds are compounds of formula III

(III)

CS 271 490 B2 in which

X, R _2; R ₃ . 22 ^{and R} 23, the meanings given are advantageous when the grouping of formulas ^R 130 - O - N - C - CO ^R 101 - ^R 21

- c - CO -

^R 22 N 1 - 0 - C - CO - NH ₂ -C - what - ^R 23

present in syn-form, ie · in Z-form, or in mixtures where the syn-form predominates ·

Yet another preferred group of compounds are compounds of formula IV

in which ’

X, R ₂ and R ₃ are as defined above ·

Also preferred are compounds of formula V

(IN)

CS 271 490 B2 in which

X, R ₂ , R ₃ and R ₁₂₀ are as defined above. Examples of salts of the compounds of formula I include alkali metal salts, such as sodium salt and potassium salt, ammonium salts, alkaline earth metal salts, such as salt. calcium, salts and organic bases; such as salts with amines (for example salts with N-methylpiperidine; procaine, dibenzylamine, N, N-dibenzylethylenediamine, alkylamines or dialkylamines), ⁱⁿ As 1 amino acid amine salts such as eoli β arginine or lysines.

Compounds of formula (I) ^: As well as their eoli and easily hydrolyzable esters ee can easily hydrate · hydratacl may occur due to the manufacturing process or the originally anhydrous product may hydrate due to its hygroscopic properties ·

The novel acyl derivatives of the formula I can be prepared according to the invention by reacting from the ester of the formula XIX r ₂ h

COOR in which

R 1, R ₂ and R ₃ are as defined above and

R is a protecting group of a carboxyl group which is used in the chemistry of cephaleporins and which can be readily cleaved under mild conditions, for example a tert-butyl group; p-nitrobenzyl or allyl, cleaves the protecting group R; a preferred p-nitrobenzyl group R by hydrolysis in the presence of sodium air, a tert-butyl group reacting with trifluoroacetic acid in the presence of anisole; or an allyl group by palladium catalyzed transallylation in the presence of a sodium or potassium salt of 2-ethylhexanoic acid / thereafter, optionally recovering the product obtained as a salt; hydrate or salt hydrate ·

Conversion of esters of formula (XIX) to the free carboxylic acid by removal of the ester protecting group according to the invention is accomplished using reagents compatible with the used ester protecting group. Oako ester protecting groups R can be used esters which can be easily converted to the free carboxylic group under mild conditions; examples of which can be exemplified by tert-butyl, p-nitrobenzyl; a benzhydryl group; allyl group, etc. · The abovementioned easily hydrolyzable ester moieties may also be used. to room temperature in a solvent such as dimethylformamide (aqueous); the tert-butyl group is removed by treatment with trifluoroacetic acid in the presence of aiol at a temperature of 0 ° C to room temperature; e or without an additional solvent such as methylene chloride: the allyl group ss removes palladium (O) catalyzed transallylation in the presence of sodium or potassium eoli 2-ethyl

Hexanoic acid (cf., for example, 3-Org. Chem. 1982, 47, 587).

□ if R1 in the compounds of formula XIX contains a protected amino group, it can be cleaved under the conditions described in variant b) below.

The preparation of the salts or hydrates of the compounds of the formula I or the hydrates of these salts by the process according to the invention can be carried out in a manner known per se, for example by reacting the carboxylic acid of the formula I or a salt thereof with organic solvent (for example in ethanol, methanol, acetone, etc.) · The eoli formation temperature is not critical and is generally around room temperature, but can also be operated at a temperature slightly above room temperature or at a temperature slightly below room temperature. room, for example in the range from 0 ° C to +50 ° C.

Hydrates are usually produced automatically as a result of the manufacturing process or as a result of the hygroscopic properties of the initially anhydrous product. In the deliberate manufacture of a hydrate, the wholly or partially anhydrous carboxylic acid of the formula The starting materials of the formula (XIX) can be prepared in various ways, for example by:

a) reacting a compound of formula XVIII ^R 2 H

(XVIII) in which

R, X and Rg are as defined above, and the carboxy and / or amino group may be present in protected form.

with an acid of the general formula

RjCOOH in which

R 1 is as defined above, or with a reactive functional derivative thereof, or

b) in order to prepare a compound of formula I in which R 1 contains an amino group as a substituent, or an ester thereof or a salt thereof, cleaves the amino protecting group in the substituent R 1 q of a compound of formula XX

COOR

CS 271 490 B2 in which

R, R 2 and R ₃ ^R 10 or are as defined above and represent an acyl group containing an amino protecting group,

c) reacting a compound of formula XXI * to produce compounds of formula I wherein X is -OCO- or -SCO-

COOR (XXI) in which

R, Κχ are as defined above, and Y is a leaving group.

with a carboxylic acid salt of the general formula

R ₃ COQH wherein ^R 3

Q is as defined above and represents an oxygen atom or an sulfur atom, or is

d) or for the preparation of -OCONH-, reacting a compound of formula XXII of formula I wherein:

X represents a -OCO- group in which

COOR (XXII)

8Θ ^R 1 ^{and R} 2 ^are as hereinbefore defined, with a compound of formula

R ₃ COZ,

R ₃ NHCO 2 or R ₃ NC0 in which

CS 271 490 B2

Rg is as defined above and

Z is an acyl activating group, or is

e) reacting a compound of formula XXIII to produce a compound of formula I wherein X is -NHCO-

R ₂ h

(ШП)

COOR in which

R @ 1, R @ 1 and R @ ₂ are as defined above and

A is an anion of an acid, with an acid of the general formula

R ₃ -COOH wherein

R ₃ meaning stated above.

The reaction of the compounds of the formula XVIII with acids of the formula R ^COOH or with their reactive functional derivatives according to variant a) can be carried out in a manner known per se. The carboxy group in the compounds of formula XVIII is protected by an R group, for example by esterification to form an easily cleavable ester, such as a tert-butyl ester, a silyl ester (e.g., trimethyleilyl ether). Further, the carboxy group can be protected by salt formation of β with an inorganic or tertiary organic base such as triethylamine. The amino groups present in the R1 groups can be protected. Suitable protecting groups are, for example, protecting groups which are cleavable by acid hydrolysis (for example tert-butoxycarbonyl or trityl) or by hydrolysis under basic conditions (for example trifluoroacetyl). Preferred protecting groups are chloroacetyl, bromoacetyl and iodoacetyl, especially chloroacetyl. These latter ee protecting groups can be cleaved by thiourea. The 7-amino group in compounds of formula XVIII may be protected, for example by a silyl protecting group, such as a trimethyleilyl group.

Příklady examples of reactive functional derivatives of acids of the formula R RCOOH include halides (ie chlorides * bromides and fluorides), azides, anhydrides, especially mixed anhydrides with strong acids, reactive esters (eg N-hydroxysuccinimidesters) and amides (eg imidazolides) .

In the reaction of a 7-amino derivative of a compound of formula XVIII θ with an acid of formula R ^COOH or a reactive functional derivative thereof, for example 8 with a free acid of formula RjCOOH, the above ester of a compound of formula XVIII can be reacted in the presence of a carbodiimide As in ethyl acetate, acetonitrile, dioxane, chloroform, methylene chloride, benzene or dimethylformamide, then the ester 9 group can be cleaved off. In the above-mentioned reCS 271 490 B2, the oxazolium salts (e.g., N-ethyl-5-phenylisoxazolium-3'-sulfonate) may be used instead of carbodilmides.

In another embodiment, a salt of an acid of formula XVIII (e.g., a trialkylammonium salt such as a triethylammonium salt) is reacted with a reactive functional derivative of an acid of formula R 3 COOH as mentioned above in an inert solvent (e.g. one of the above solvents).

According to another embodiment, the acid halides, preferably chlorides, of the acid of the formula R1COOH and the amine of the formula XVIII are reacted. This reaction is preferably carried out in the presence of an acid-binding agent, for example in the presence of an aqueous alkali, preferably sodium hydroxide, or in the presence of an alkali metal carbonate such as potassium carbonate, or in the presence of a lower alkylamine such as triethylamine. Oako solvents preferably use water, optionally in admixture with an inert organic solvent such as tetrahydrofuran or dioxane. The reaction may also be carried out in an aprotic organic solvent such as dimethylformamide, dimethylsulfoxide or hexamethylphosphoric triamide, since the silylated compounds of formula XVIII are used, then the reaction is carried out in an anhydrous medium.

Preferred acylece alternatives in which the amino group present in R 1 may not be protected are the use of a 2-benzthiazolylthioester or 1-hydroxybenzotriazoleeter acid of formula R 3 COOH. For example, the 2-benzthiazolylthioaater may be reacted with a compound of formula XVIXI in an inert organic solvent such as a chlorinated hydrocarbon such as methylene chloride, acetone, ethyl acetate, or a mixture of such solvents and water with 1-hydroxybenzotriazol esters. R 1 COOH e 1-hydroxybenzotriazole and carbodiimide, in particular N, N'-dicyclohexylcarbodiimide or N, N'-dilsopropylcarbodiimide in an inert organic solvent, in particular methylene chloride, dimethylformamide, tetrahydrofuran, acetonitrile or ethyl acetate.

The reaction of the 7-amino derivative of formula XVIII with an acid of formula R 3 COOH or a reactive functional derivative thereof ee can usually be carried out at a temperature between about -40 ° C and * 60 ° C, for example at room temperature.

The cleavage of the amino group in the substituent R1 of the compound of formula (XX) according to variant b) yields the corresponding esters of formula (XIX) containing a compound of formula (XIX). Suitable amino protecting groups are those used in peptide chemistry such as alkoxycarbonyl, for example tert-butoxycarbonyl, etc., substituted alkoxycarbonyl, for example trichloroethoxycarbonyl, etc., substituted aralkyloxycarbonyl, for example p-nitrobenzyloxy. an aralkyl radical such as a trilyl acylamino / hydroxy group (or a haloalkanoyl group, such as a chloroacetyl group, a bromoacatyl group, an iodoacetyl group or a trifluoroacetyl group).

Preferred protecting groups are t-butoxycarbonyl (t-BOC) and trityl.

Amino protecting groups can be cleaved by acid hydrolysis (e.g., tert-butoxycarbonyl or trityl) by nobo hydrolysis under basic conditions (e.g., trifluoroacetyl). The chloroacetyl group, bromoacetyl group and iodoacetyl group are removed by treatment with thiourea.

Amino protecting groups which are cleavable by acid hydrolysis are preferably removed by treatment with an alkanecarboxylic acid which can be halogenated. In particular, formic acid or trifluoroacetic acid is used. The acid hydrolysis is generally carried out at ambient temperature, although ee may be carried out at a temperature that is slightly higher or slightly below the ambient temperature (e.g., at a temperature in the range of about 0 ° C).

CS 271 490 B2 up to +40 ° C). Protective groups which are detachable under basic conditions are generally hydrolyzed with dilute aqueous caustic solution at a temperature of 0 ° C to +30 ° C. The chloroacetyl group, the bromoacetyl group and the iodoacetyl group, which groups are used as protecting groups, can be cleaved off using thiourea in an acid, neutral or alkaline medium at a temperature of about 0 ° C to 30 ° C.

The reaction of the compounds of the formula XXI with the salts of the compounds of the formula R ^COQH according to variant c) is carried out in a solvent which does not contain hydroxyl groups, such as dimethylformamide, methylene chloride or Ν, dim-dimethylacetamide. Other solvents which do not contain hydroxyl groups can also be used. Suitable salts for compounds of the formula R ₃ C0QH e.g. sodium salt, potassium salt, cesium salt, tetrabutylamonlová tetramethylamonlová salt or salt thereof. Hal 'represents a halogen atom, preferably a bromine atom or an iodine atom. The reaction may also be carried out in aqueous bicarbonate with any of the salts of the compound of formula RqCOQH mentioned. The reaction is preferably carried out at temperatures of about 0 to 80 ° C, preferably at room temperature.

The reaction of compounds of formula XXII with compounds of formula R 2 CO 2, R ₄ NHCO 2 or R ₃ NCO according to variant d) ee is carried out in a manner which is further illustrated by reaction schemes II and V ·

The reaction of the compounds of formula XXIII and their esters and their salts with acids of the formula R ₃ COOH according to variant e) is illustrated below in Scheme IV.

In the following reaction schemes, compounds of formula X in Scheme I, compounds of formula XIII in Scheme III, compounds of formula XVI in Scheme IV, and compounds of formula XVII in Scheme V, R ₃ (as defined above) may be hydroxyaryl or its alkanoyl ester In the case of alkanoyl esters, the conversion to the corresponding hydroxyaryl radicals can be carried out using an alcohol and a base, preferably methanol and sodium bicarbonate.

The following reaction schemes combine to further elucidate the processes for producing the compounds of the present invention.

S c hé m a I

CS 271 490 B2

In the above scheme, the substituents R p R ₂ ^{and R} 3 are as defined above.

In the above reaction scheme, according to the choice of the carboxyl function (R) protecting group and the halogen atom used, the double bond in the cefsm ring can occupy the position A 2 or Δ 2 relative to the sulfur atom. The resulting mixed product can, if desired, be purified to give only the desired isomer by conversion to the sulfoxide (IX), which is then subjected to reduction or can be purified by separation of the two components.

Scheme I '

VI - VII + VIII

A compound of formula (VI), which is known or is produced in an analogous manner to known compounds (cf., for example, U.S. Patent No. 4,406,699 and U.S. Patent No. 4,266,049), is reacted with a salt of the selected carboxylic acid. shall be carried out according to the procedure described in shors for variant (c) ♦

VII-> X

The compound of formula (VII) is then deprotected as described for the process of the invention to give the carboxylic acid of formula (X) or a mixture thereof with the corresponding Δ2-isomer.

VIII-I-IX

When the double bond isomerized, the compound of formula (VIII) is oxidized, for example, by a reaction such as meta-chloroperbenzoic acid, in a solvent such as methylene chloride at a temperature of about -20 ° C to +40 ° C, preferably at about 0 ° C.

IX- ^ VII

The compound of formula (IX) is then reduced to the desired end product of formula (VII) using various reactions, for example by treatment with phosphorus trichloride in dimethylformamide or trifluoroacetic anhydride in the presence of sodium iodide in acetone / methylene chloride. in the range of about 0 ° C to -20 ° C, and is preferably about 0 ° C.

Scheme II

(VIII) (VIII)

CS 271 490 Q2

In the above scheme, the general substituents R, Rj, Rg and Rg are as defined above.

Scheme II

XI — VII * VIII

A compound of formula XI which is known or is prepared according to the procedures described in ournal of Antibiotics, 1981, 34, 1300, is reacted with a compound of formula R 1 -CO-Z in which Z is an acyl activating group. A preferred activating group Z is a halogen atom and a group of formulas

CH ₃

wherein it is an alkyl group having 1 to 3 carbon atoms.

The reaction is carried out in a solvent such as methylene chloride at a temperature from about -20 ° C to 100 ° C, preferably at about 25 ° C. The double bond in the cephem ring of the resulting product may, due to iaomerization, occupy a position of Δ 3 or Z к 2 relative to the sulfur atom. The mixed product can be purified to give only one isomer as described above for Reaction Scheme I by conventional separation or oxidation with a peracid to give the sulfoxide (IX) and to be liberated by reduction of this compound to give the protected ester (VII).

CS 271 490 B2

Scheme III

(1И) (ХГI 1) where

R p R g and R 3 are as defined above and

Ac is CH 2 CO-.

Scheme III

XII—> XIII

The compound of formula (XII) or a salt thereof is preferably reacted with aqueous sodium bicarbonate by a compound of formula (ASCO-Rg) wherein R ₃ is as defined above and A is a cation, preferably eodic, at a temperature of from 25 to 100 ° C. , preferably at a temperature of 40 to 60 ° C, to give the desired product (XIII).

The compound of formula (XIII) can also be prepared according to Reaction Scheme I, wherein the selected cyeeline is a cyeeline of the formula R ^COSH.

Scheme IV

H

COOH

(XIV)

(XV)

CS 271 490 B2 i.

whereas

A, A, Rp, Rg and Rg are as defined above.

Scheme IV

XII XIV

The compound of formula (XII) is reacted with an azide (sodium or potassium) salt in an aqueous base such as sodium bicarbonate at a temperature of from 25 to 100 ° C, preferably at 40 to 60 ° C.

XIV — XV

The azide (XIV) is reduced by hydrogen in the presence of a catalyst such as palladium or platinum, or preferably by metal tin in the presence of hydrochloric acid at a temperature of 0 to 50 ° C, preferably at a temperature of 20 to 35 ° C.

XV — xvi

The amide bond is formed by standard methods of coupling an amine with an acid. As is customary in peptide chemistry (cf., for example, Syntheeis, 1972, p. 463), preferably using dicyclohexylcarbodiimide and N-hydroxybenzotriazole or triphenylphosphine and diaryldisulphide (for example 2,2'-dibenzothiazolyldisulphide) at a temperature of about 0 to 50 ° C, preferably at a temperature of 20 to 35 ° C, and in an inert solvent, for example dimethylformamide or dimethyl sulfoxide.

Scheme V

OH

o o

th

OCNHR ₃

COOIř while

R, Rp, Rg and Rg are as defined above.

(XVII)

CS 271 490 B2

Scheme V

XI - XVII

The compound of formula (XI) ee reacts with a compound of formula (O) -C (NR ₃₎ or ZCONHR (4) optionally in the presence of a base (e.g. pyridine, picoline or lutidine), wherein R ₃ is as defined above and Z is an acyl activating group; The reaction is carried out in a solvent, for example methylene chloride, at a temperature of about 0 ° C to 60 ° C, and preferably at 25 ° C. If desired, a protecting group is removed from the compound of formula (XVII).

according to the procedure described in Reaction Echema I. Compounds of general formula I containing groupings of general formulas ^R 130 - 0 - N-C-CO- AND 1 ^R 101 í ^0R 21 - c - CO - a - * 22 N - 0 M - C - CO-NHR _1g -c-what- * 23

(erov. from above) are preferably syn-forms. Such syn-forms ae can be obtained using starting materials in which this syn-form is already present. If desired, the obtained emyne / anti-form can be separated into the corresponding anti-forms by conventional means, for example by recrystallization or chromatographic methods. using a suitable solvent or solvent mixture ·

The compounds of formula (I), their salts and esters, as well as the hydrates thereof, have antibiotic but in particular bacterial activity and can be used as a means of combating bacterial infections (including urinary tract infections and respiratory tract infections) in mammals such as dogs, cats, equines etc. · as well as humans · These compounds have an activity against a broad spectrum of both gram-negative and gram-positive bacteria.

The in vitro activity of the compounds of the invention against a variety of Gram-negative and Gram-negative bacteria, expressed as the minimum inhibitory concentration in micrograms / ml and determined using the agar digestion method, agar dilution method and broth dilution method, is as follows:

Compound A: (6R-) / 6c (1,7 / 3 (Z)) - 7 - [(2-amino-4-thuazolyl) (methoxyimino) acetyl] amino] -3 - [(3, 4-Dihydroxybenzoyl-allyl-methyl-6-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid

Compound B: (6R-) 6,7- (Z) // -3- [3,4-bis (acetyloxy) benzoyl] oxy (methyl) -7- (2-amino-4) monosodium salt -thiazolyl // (2-amino-2-oxethoxy) imino (acetyl) amino] -8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid

Compound C: (6R-) 6or, 7y (Z) H-7 - [(2-amino-4-thiazolyl) -1-carboxy-1-methyl] disodium salt18

CS 271 490 B2

sthoxy) imino (acetyl) amino] -3 - [(3,4-dihydroxybenzoyl) oxy] methyl] -8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-en-2 -carboxylic acids Compound 0: Monosodium salt / 6R / 6 <X 7Д ^(Rx) // - 7 - //// (4-athyl-2,4-dioxo-l-piperazinyl) carbonyl / amino / phenylacetyl / amino / -3 - [(3,4-dihydroxy) benzoyl (oxy) methyl] -8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-an-2-carboxylic acid

Table 1

Minimum in vitro inhibitory concentration (yug / ml) (diffusion method on agar wells)

Microorganism

P. aeruginosa

P. vulgaris 101N

E. coli 1269B

K. pneumonia

S. marcescans

S. aureus 1059B Compound A

0.98

0.06

0.12

0.49

0.98

7, no

Minimum inhibitory concentration in (agar dilution method)

Microorganism

E. coli

K. pneumoniae A

E. cloacae 9570A

E. cloacae P99

P. vulgaris ATCC 6380

P. mirabilis

S. marscescens SM

P. aeruginosa

P. aeruginosa 185 / H

P. aeruginosa Stone

S. aureua Giorgid

S. aureus Smith

S. aureus

S. aureus 10598

S. aureus ATCC 25923

Table 2 vitro (/ Ug / ml) Compound В

0,031

0.125

0.016

0,063

Table 3

Minimum In vitro Inhibitory Concentration (/ Ug / ml) (Broth dilution method)

Microorganism compound

C

P. aeruginosa

P. aeruginosa Stone 130 P. aeruginosa ATCC 27853

P. aeruginosa 8710

P. aeruginosa 503-56

0.1250.25

0.1250.5

320.5

0,0630,125

0.254

CS 271 490 B2 continued Table 3

Microorganism AND Compound C 0 P. aeruginosa 8780. 0.5 <0.008 0.016 P. aeruginosa 6148B 2 0,031 0.125 P. aeruginosa 765 4 8 128 P. aeruginoea 185 / H 0.016 0,031 0.125 P. aeruginosa 19736 4 2 4 P. aeruginoea 5700 8 2 4 P. aeruginosa K77 / WT 1 0.63 0.25 P. aeruginoea K77 / 61 0.25 <0.008 0.016

To combat bacterial infections, a compound of the invention, i.e., a compound of formula I, or a hydrolyzable ester thereof, or a pharmaceutically acceptable salt or corresponding hydrate thereof, can be administered to a mammal in an amount of about 5 mg / kg / day to about 500 mg / kg / day. preferably in an amount from about 10 mg / kg / day to about 100 mg / kg / day, particularly preferably in an amount from about 10 mg / kg / day to about 55 mg / kg / day.

Many of the applications that are currently contemplated in the treatment of infectious diseases of penicillins and cephalosporins are also useful for the compounds of the present invention. Thus, administration can be carried out, for example, orally, such as by means of tablets or capsules, parenterally, for example, intravenously or intramuscularly. Also enterally.

The cephalosporin derivatives obtained by the process of the present invention can be used as medicaments, for example in the form of pharmaceutical preparations containing these compounds in admixture with a pharmaceutically acceptable carrier. The carrier may be organic or inorganic inert carrier materials suitable for enteral or parenteral administration, such as water, gelatin, acacia, lactose, starch, magnesium stearate, talc, vegetable oils, polyalkylene glycols, vaseline, etc. Pharmaceutical compositions may be present. in solid form, for example in the form of tablets, dragees, suppositories or capsules, or in liquid form (for example in the form of solutions, suspensions or emulsions). The pharmaceutical preparations are optionally sterilized and / or may contain adjuvants such as preservatives, stabilizers, wetting or emulsifying agents, salts for varying the osmotic pressure, anesthetics or buffers. The pharmaceutical preparations may also contain other therapeutically valuable substances. The carboxylic acids of the formula I, their salts and hydrates, are particularly suitable for parenteral administration and are preferably prepared in the form of lyophilisates or dry powders intended for dilution by conventional means, such as water or isotonic sodium chloride, with cosolvents such as propylene glycol. Easily hydrolyzable esters or ethers of formula I are also suitable for enteral administration.

In the following examples, e indicates chemical shifts in the NMR spectrum as δ values, and the band positions in the infrared (IR) band are reported in ии *. .

Example 1

Preparation of 1,1-Dimethylethyl ether (6R- [6,7,7] (Z)] - 3- (acetoxymethyl) -7-amino-8-oxo-5-thia-1-azabicyclo [4.2.0] of oct-2-ene-2-carboxylic acid

To 200 ml of ice-cooled dioxane was added 10 ml of concentrated sulfuric acid, followed by 21.78 g (80.0 mml) of 7-aminocephalosporanic acid in one portion. The mixture is cooled to -50 ° C and 50 ml of liquid isobutylene (especially condensed under argon atmosphere) are added. The pressure vessel was sealed and the mixture was agitated overnight (at about 10 to 50 psig). The mixture was then cooled to -50 ° C and the pressure tube opened.

CS 271 490 82

The contents were poured slowly into a solution of sodium bicarbonate in water (100 g NaHCO 3 in 1500 mL water) with stirring. The solution obtained is extracted three times with 500 ml of ethyl acetate each time, and the combined organic extracts are washed with brine and dried over magnesium sulphate. Removal of the solvent under reduced pressure gave a yellow-orange oil. Trituration of this oil with petroleum ether (b.p. 30-60 ° C) afforded a solid product which was filtered off and dried. 17.1 g (65% of theory) of product are obtained, which product is used without further purification.

NMR Spectrum (CDCl3):

1.55 (s) 9H (χ-bu). 2.07 (s) 3 H (OAc). 3.33. 3.54 (dd, 3 to 10 Hz) 2H _(CH2 S), 4.75, 5.02 (dd, 3-20 Hz), 2H (CH ₂ 0), 4.72 (d, 3-10 Hz 1 H (C 6), 4.91 (d, 3 → 10 Hz) 1 H (C 7).

Example 2

Preparation of [6R- / 6oC, 7D (Z)] - 3- (acetoxymethyl) -7 - [(methoxyimino)] - (2-tritylamino) -4-thiazolyl] acetyl / amino] 1,1-dimethylethyl ester -8- oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid

To a stirred suspension of 48 g (0.1 mol) of 2- [2- (tritylamino) thiazol-4-yl] -2-methoxyiminoacetic acid in 750 ml of anhydrous dichloromethane is added 13.94 ml (0.1 mol) of triethylamine and the mixture is stirred mechanically until a clear solution is formed (approximately 45 minutes). After complete dissolution, 20.6 g (0.1 mol) of Ν, d-dicyclohexylcarbodiimide and 13.5 g (0.1 mol) of 1-hydroxybenzotriazole were added and the mixture was stirred at room temperature for 2 hours. Then, a solution of 32.8 g (0.1 mol) of 1,1-dimethylethyl (6R- / 6X, 7R (Z)) - 3- (acetoxymethyl) -7-amino-8, 1,1-dimethylethyl ester is added to the reaction mixture. oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid (from Example 1) in 250 mL of anhydrous dichloromethane and the resulting solution was stirred at room temperature for 16 hours. The mixture is filtered and the filtrate is washed successively with 2 x 250 ml, i.e. 500 ml of saturated aqueous sodium bicarbonate solution and 2 x 250 ml, i.e. 500 ml of saturated aqueous sodium chloride solution, and dried over sodium sulfate to remove the crude product as a gum. The oil was purified by preparative liquid chromatography using methylene chloride containing 0-10% ethyl acetate as eluent to give 57.2 g (76%) of a pale yellow solid.

NMR Spectrum (CDCl3):

1.54 (s) 9H (χ-bu), 2.09 (s) 3H (OAc), 3.35, 3.56 (dd, 3-18 Hz) 2H (CH ₂ S), 4.08 ( s) 3 H (0CH _3), 4.86, 5.04 (dd, 3-14 Hz), 2H (CH ₂ 0), 5.04 (d, 3-6 Hz) 1H (C6),

5.93 (dd, 3-6 Hz, 3-10 Hz), 1H (C7), 6.71 (s) 1H (thiazole), 6.91 (d, 3-10 Hz) 1H (NH), 7 .08 (s) 1H (NH), 7.30 (z) 15H (CPhg). ^;

Example 3

Preparation of [6R-] 6oc, 7/3 (Z) // -3- (iodomethyl) -7- [(methoxyimino)] - (tritylamino) -4-thlazolyl (acetyl) amino] 1,1-dimethyl ester 8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid

To a stirred solution of 23.8 g (0.0316 mol) of 1,1-dimethylethyl ester / 6R- / 6o (.7 / 3 (Z) // - 3- (acetoxymethyl) -7 - // (methoxyimino) / 2- (tritylamino) -4-thiazolyl (acetyl) amino] -8-oxo-5-thia-1-azablicyclo [4.2.0] oct-2-ene-2-carboxylic acid (from Example 2) in 250 mL of anhydrous dichloromethane (distilled from phosphorus pentoxide) at room temperature is gradually added in three portions at ten minute intervals of 2.0 ml (0.014 mol), 2.0 ml (0.014 mol) and 1.5 ml (0.0105 mol) of iodotrimethylsilane. After the last addition, the mixture was stirred at room temperature for 2 hours and then 0.5 ml (0.0035 mol) was added

1.3 equivalents) of iodotrimethylsilane. After the reaction mixture was stirred for an additional 30 minutes, the solvent was evaporated at 0 ° C under reduced pressure (on an ice bath).

CS 271 490 B2

The 21% product was dissolved in 250 mL of ethyl acetate (previously cooled to 0 ° C). The ethyl acetate solution is washed (with all the washings previously cooled to about 0 ° C) successively with 3 125 125 ml of 375 ml of cold aqueous sodium thiosulfate solution, 125 ml of saturated aqueous sodium bicarbonate solution, 2 x 125 ml of saturated aqueous solution. sodium chloride and dried over anhydrous sodium sulfate. The solvent was removed at 0 ° C under reduced pressure, and the obtained gummy product was immediately purified by preparative liquid chromatography (solvent system: ethyl acetate, n-hexane and methylene chloride 1: 4: 6) to give 15.4 g ( 59% of theory) of the desired iodomethylacaphalosporin in the form of a light yellow solid product. The unreacted starting material (5.0 g) is also recovered, increasing the yield relative to the reacted starting material to 75%.

NMR Spectrum (CDCl3):

1.66 (s) 9H (t, -bu), 3.51, 3.76 (dd, 0-20 Hz), 2H (CH3S), 4.11 (s) 3H (OCH3), 4.30,

4.44 (dd, 3 · 10 Hz) 2H (CH ₂ ), 5.03 (d, 3.6 Hz) 1H (C 6), 5.89 (dd, 3 - 6 Hz, 3 · 10 Hz) 1H ( C7), 6.72 (m) 2H (thiazole, NH), 7.02 (s) 1H (NH), 7.30 (b) 15H (CPh ₃ ).

IR (KBr): 3385, 3285, 1787, 1717, 1681, 1524, 701.

Example 4

Preparation of [6R- / 6oc, 7β (2)] - 3 - [(3,4-bis (acetyloxy) benzoyl) oxy] methyl 1,1-dimethylethyl ester-8-oxo-7 - /// 2 - ((triphenylmethyl) amino) -4-thiazolyl] (methoxyimino) acetyl (amino) -5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid

To a solution of 6.98 g (0.085 mol) of 1,1-dimethylethyleatar (6R) 6tf, 7tf (Z) - 3- (iodomethyl) -7 - /// (methoxyitnino) -2 - [(triphenylmethyl) amino] -4-thiazolyl (acyl) -amino] -8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid (from Example 3) in 100 mL of absolute dimethylformamide a stirred solution of 2.29 g (0.086 mol) of (3,4-diacetoxy) benzoic acid sodium salt in 100 ml of absolute dimethylformamide (dried over 1 hour with molecular sieves 4, 10 * 10 &lt; -1 &gt; m) was added dropwise under argon at ambient temperature. . The reaction mixture was stirred for 3 hours and then the solvent was removed under reduced pressure. The oily residue was dissolved in 100 ml of a 9: 1 mixture of methylene chloride and ethyl acetate and transferred to a short column of silica gel. Fractions containing the desired product were combined to give 7.25 crude product. Further purification using preparative chromatography (solvent system: 5: 4 n-hexane: ethyl acetate) gave 3.9 g (42%) of a cream colored solid.

NMR Spectrum (CDCl3):

1.44 (δ) 9H (δ -bu), 2.27 (s) 3 H (OAc), 2.29 (e) 3 H (OAc), 3.63, 3.73 (dd, 3-10 Hz) 2H ( CH ₂ S), 3.81 (s) 3 H (OCH ₃ ), 4.91, 5.19 (dd, 3 · 16 Hz) 2H (CH ₂ O), 5.13 (d, 3 ·

Hz) 1H (C6), 5.99, 6.01 (dd, 3-6 Hz, 3-6 Hz) 1H (C7), 6.68 (b) 1H (thiazole),

7.2-7.4 (m) 15H (CPh ₃ ), 7.44 (d, 3-9 Hz) 1H (Ar), 7.84 (s) 1H (Ar), 7.91 (d, J) 9 Hz) 1H (Ar), 8.86 (s) 1 H (NH), 9.59 (d, 3-10 Hz) 1 H (NH).

IR spectrum (KBr technique): 3300, 1778, 1772, 1682. UV spectrum (ethanol): λ _roax 234 ( _δ · 36500) Mass spectrum: m / z 932 (Μ + H).

Example 5

Preparation of (6R-) - [6,7- (2)] - 3 - [(3,4-dihydroxybenzoyl) oxy] methyl] -8-oxo-7 - (// 2-) 1,1-dimethylethyl ester (triphenylmethyl) amino] -4-thiazolyl] (methoxyimino) acetyl] amino) -5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid.

CS 271 490 B2

Using the procedure and conditions described in Example 4, 374 mg (2.12 mmol) of 3,4-dihydroxybenzoic acid sodium salt in 80 ml of dimethylformamide were added to 1.65 g (2.0 mmol) of 1,1-dimethylethyl ester (6R-). 6 <X, 7 (J) (3-) (iodomethyl) -7 - [(methoxyimino) -2 - [(triphanylmethyl) amino] -4-thiazolyl] acetyl] amino] -8-oso- Of 5-thia-1-azabycyclo [4.2.0] oct-2-ene-2-carboxylic acid in 50 ml of dimethylformamide, the reaction mixture is stirred for 1.5 hours, and then 1.71 g (50% of theory) are obtained. ) after chromatography on silica gel (using methylene chloride / methanol 99: 1) as a cream colored solid ·

NMR Spectrum (CDCl3):

1.22 (s) 9H (£ -Bu), 3.30, 3.54 (dd, 3-16 Hz) 2H (CHGS), 3.82 (s) 3H (0CH _3), 4.04 br s 1 H (OH), 5.07 (d, 3-6 Hz) 1H (C 6), 5.23, 5.32 (dd, 3-12 Hz), 2H (CH ₂ O), 5.95 (dd) 1.6 Hz, 3-10 Hz) 1H (C7), 6.77 (s) 1H (thiazole), 6.87 (d, 3.6 Hz) 1H (Ar), 7.30 (s) 15H (CPh ₃ ), 7.49 (m) 3 H (Ar).

IR (KBr): 3290, 1791, 1690, 1522, 702;

UV spectrum (ethanol) _max 260 (? 23,200).

Mass spectrum: m / z 848 (M + H).

Example 6

Preparation of [6R- (60), 7/2 (Z)] - 3 - [(2,3-dlhydroxybenzoyl) oxy] methyl] -8-oxo-7 - (2) - (1,1-dimethylethyl ester) triphenylmethyl) amino (4-thiazolyl) (methoxyimino) acetyl (amino) -5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid

Following the procedure and conditions described in Example 4, 35.3 mg (0.20 mmol) of 2,3-dihydroxybenzoic acid sodium salt in 5 ml of dimethylformamide were added to 157 mg (0.19 mmol) of 1,1-dimethyl ethyl ester (6R-). 60C, 7/3 (Z) H-3- (iodomethyl) -7 - [(methoxyimino) -2 - [(triphenylmethyl) amino] -4-thiazolyl] acetyl / amino] -8-oxo-5 -thia-1-azabycyclo [4.2.0] oct-2-ene-2-carboxylic acid in 5 ml of dimethylformamide, the reaction mixture is stirred for 2 hours and 15 minutes, after chromatography on silica gel (using methyl chloride) and 0-10% ethyl acetate as solvent system) gave 111 mg (69%) of an off-white solid.

NMR Spectrum (CDCl3):

1.59 (s) 3 H (t-bu), 3.48, 3.68 (dd, 3 H = 20 Hz) 2H (CH ₂ S), 4.07 (s) 3 H (OCH ₃ ), 10 (m) 2H (6, l / 2 CH ₂ 0), 5.48 (d, 3 «14 Hz) 1H (l / 2 CH ₂ 0), 5.99 (m) 1H (C7), 6.67 ( broad з) 1H (OH), 6.72-6.88 (m) 3H (thiazole, 2 Ar), 7.02 (broad s) 1H, 7.16 (dd, 3 → 10 Hz) 1H (NH) 7.28 (s) 15H (CPh ₃ ), 10.71 (s) 1H (OH).

IR (KBr) 3400, 1789, 1722, 1688, 700.

UV spectrum (ethanol): λ _max 240 nm (· 33,900).

Mass spectrum: m / z 848 (M + H).

Example 7

Preparation of [6R- (60C) 7,7 (Z)] - 3 - [(3,4,5-tris (acetyloxy) benzoyl) oxy] -methyl] -7- (3-methoxyimino) 1,1-dimethylethyl ester [2- (triphenylmethyl) amino] -4-thiazolyl (acetyl) amino] -8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid

Using the conditions described in Example 4, 59 mg (0.195 mmol) of 3,4,5-triacetoxybenzoic acid sodium salt in 2 ml of dimethylformamide were added to 160.8 mg of 1,1-dimethylethyl ester (6R- (60C, 7β (Ζ))). [3- (iodomethyl) -7 - [(methoxyimino) -2- [(triphenylmethyl) amino] -4-thiazolyl] acetyl] amino] -8-oxo-5-thia-1-azabicyclo [4], 2, O / oct-2-ene-2-carboxylic acids in 2 ml dimethylformamide, the reaction mixture is stirred for 2 hours and after purification by chromatography on silica gel (using methylene chloride 9 containing 0-15% ethyl acetate),

CS 271 490 В2 gives 64.1 mg (33% of theory) of a cream colored solid.

NMR Spectrum (CDCl3):

1.52 (β) 9 H (^ tBu), 3.37, 3.59 (dd, 3 to 18 Hz) 2H _(CH2 S), 4.06 (β) 3 H (OCHg), 5.02 ,

5.38 (dd, 7.3 Hz) 2H _(CH2 O), 5.05 (d, 3-6 Hz) 1H (C6), 5.95 (dd, 3 »6 Hz, 10 Hz · □) 1H (C7), 6.70 (d, 3-10 Hz) 1H (NH), 6.72 (b) 1H (thiazole), 6.98 (s)

1H (NH), 7.29 (s) 15H (CPhg), 7.77 (b) 2H (Ar).

IR (KBr): 3340, 1788, 1722, 1690, 701.

UV spectrum (ethanol) λ _max 235 nm (Z * 32 750).

Mass spectrum: m / z 990 (M + H).

Example 8

Preparation of [6R- (6 *, 7 ') (Z)] - 3 - [2- (3,4-dihydroxyphenyl) acetoxy] -methyl] -7 - [(methoxyimino)] - 2-dimethylethyl ester ((triphenylmethyl) amino) -4-thiazolyl (acetyl) -amino) -8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid

According to the procedure and conditions described in Example 4, 4.76 mg (0.400 mol) of 2- (3,4-dihydroxyphenyl) acetic acid sodium salt in 4.0 ml of dimethylformamide are added.

328.7 mg (0.400 mmol) of (6R-) - [6,7] (2) H -3- (iodomethyl) -7 - [(methoxyimino) -2 - [(triphenylmethyl)] -dimethyl-ethyl ester 1,1-dimethylethyl ester amino (4-thiazolyl) acyl / amino) -8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid in 4 mL of dimethylformamide, stirred 2 After chromatography over silica gel (using methylene chloride 8 containing 0-20% ethyl acetate), 215 mg (62% of theory) of a cream-colored solid are obtained.

NMR spectrum (perdeuterated dimethylsulfoxide):

1.44 (s) 9H (Jt-bu), 3.23-3.45 (m) 4H (CH 3 S, CH 3 CO), 3.80 (s) 3 H (OCH 3), 4.65, 4.95 (dd) 3-14 Hz) 2H (CH2O), 5.11 (d, 3-4 Hz) 1H (C6), 5.69 (dd, 3-4 Hz, 3-10 Hz) 1H (C7), 6, 46 - 6.65 (m) 3 H (Ar), 6.69 (s) 1 H (thiazole), 7.19 - 7.37 (m) 15 H (CPhg),

8.80 (9) 1 H (OH), 8.86 (β) 1 H (NH) 6.88 (s) 1 H (OH), 9.57 (d, 3-10 Hz) 1 H (NH).

IR (KBr): 3390, 1788, 1723, 1662, 702;

UV spectrum (ethanol): λ 225 nm («19900).

max

Mass spectrum: m / z 861 (M + H).

Example 9

Preparation of (6R-trans) -3 - [(3,4-bis (ashoxy) benzoyl) oxy] methyl] -8-oxo-7 - [(phenoxyacetyl) amino] -5-thia- (1,1-dimethylethyl) ester 1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid

Using the procedure and conditions described in Example 4, 2.33 g (8.95 mmol) of 3,4-diacetoxybenzoic acid sodium salt in 100 ml of dimethylformamide are added to 4.75 g (8.95 mmol) of 1,1-dimethylethyl ester ( 6R-trans) -3- (iodomethyl) -8-oxo-7 - [(phenoxyacetyl} amino) -5-thia-1-azabixyclo [4.2.0] oct-2-ene-2-carboxylic acid in 50 ml of dimethylformamide, the reaction mixture is stirred for 2 hours and after purification by chromatography on silica gel (using n-hexane / ethyl acetate 6: 4), 3.36 g (58% of theory) of a cream-colored solid are obtained.

NMR Spectrum (CDCl3):

1.56 (s) 9H (b-bu), 2.33 (s) (6H) (2 OAc), 3.41, 3.63 (dd, 3 → 18 Hz) 2H (CH 3 S),

4.60 (β) 3 H (OCH ₃ ), 5.06, 5.40 (dd, 3 → 14 Hz) 2H (CH ₂ O), 5.07 (d, 3-6 Hz 1H (C 6), 5 .97 (dd, 36 Hz, 3-10 Hz) 1H (C7), 6.98 (d, 3-8 Hz) 1H (Ar), 7.09 (d, 3-10 Hz) 1H (NH) 7.12-7.40 (m) 5H (Ar), 7.90 (α) 1H (Ar), 7.98 (d, 3-8 Hz) 1H (Ar).

CS 271 490 B2

Example 10

Preparation of monosodium eoli [6R- / 6 *, 74 (Z)] - 7 - [(2-amino-4-thiazolyl) (methoxylmino) acetyl] amino] -3 - /// (3,4-bis ( acetyloxy) benzoyl) (oxy) methyl] -8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid

A solution of 1.90 g (2.04 mmol) of (6R-) - (6), 7 (I) - (3) - (3,4-bis (acetyloxy) -benzoyl) oxy / methyl 1,1-dimethylethyl ester (-8-oxo-7 - [] 2 - [(triphenylmethyl) amino] -4-thiazolyl] (methoxyimino) acetyl] amino] -5-thia-1-azabicyclo [4.2.0] oct-2 -en-2-carboxylic acid (from Example 4) in 38 ml of absolute methylene chloride was cooled to 0 ° C in an ice / water bath and 3.8 ml of anisole was added thereto, followed by 37.5 ml of trifluoroacetic acid. The mixture was stirred at 0 ° C for 6 hours and then the solvent was removed under reduced pressure. The residual oil is taken up in 40 ml of ethyl acetate and the ethyl acetate solution is washed with 9 parts (40 ml each) of a 1% aqueous sodium bicarbonate solution. Fractions 4 to 7 containing the active product according to the UV spectrum were combined and chromatographed over a short column of reverse phase silica gel using water to remove excess sodium bicarbonate and then 25% acetonitrile in water to remove organic material. The desired fractions were combined and lyophilized to give 760 mg of crude product. The lyophilized powder was further purified by preparative reverse phase chromatography (0-30% acetonitrile) to give 580 mg (43% of theory) of a white powder.

NMR spectrum (perdeuterated dimethylsulfoxide):

2.29 (s) 3 H (OAc), 2.30 (s) 3 H (OAc), 3.26, 3.60 (dd, 3-16 Hz) 2H (CH 3 S), 3.84 (s) 3 H (OCH 8) , 4.95, 5.21 (dd, 3-10 Hz), 2H (CH ₂ 0), 4.97 (d, 3-6 Hz) 1H (C6)

5.53 (dd, 3-6 Hz, 3 → 10 Hz) 1H (C7, 6.69 (s) 1H (thiazole), 7.19 (s) 1H (NH), 7.39 (d, 38) Hz) 1 H (Ar), 7.80 (s) 1 H (Ar), 7.87 (d, 3-8 Hz) 1 H (Ar), 9.51 (d, 3 → 8 Hz) 1 Η (NH) .

IR (KBr): 3350, 1769, 1715, 1685, 1610.

Mass spectrum: m / z 656 (M + H).

UV spectrum (water): λ _max 235 nm (<£ 26,500) ·.

HRMS calcd for ^C 25 ^H ₂₃ ^N 5 ^O 5 ^S 2 (Μ ♦ H) 656.0733 found (Μ + H) 656.0732.

Example 11

Preparation of (6R-) 6Cl, 7/3 (Z) // - 7 - [(2-amino-4-thiazolyl) (methoxyimino) acetyl / amino] -3- [(3,4-dihydroxybenzoyl) monosodium salt oxy (methyl) -8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid

Using the procedure and conditions described in Example 10, 52.3 mg (0.092 mmol) of (6R-) - (6) - (6) - (3) - (3 -) - ( 3,4-dihydroxybenzoyl) oxy (methyl) -8-oxo-7 - [(2H) - [(triphenylmethyl) amino] -4-thiazolyl] (methoxylmino) acetyl] amino] -5-thia-1-azabicyclo [4] 2.0 / oct-2-ene-2-carboxylic acids (from Example 5) in 1 mL methylene chloride and 100 µL anisole in 1 mL trifluoroacetic acid, then stirred for 3 hours and 40 minutes. Purification by reverse phase chromatography (0-30% acetonitrile in water) gave 11.3 mg (32%) of a white powder.

NMR spectrum (deuterated water):

3.47, 3.73 (d, 3-18 Hz) 2H _(CH2 S), 3.98 (s) 3H (OCHg), 4.89, 5.11 (dd, 3 to 10 Hz) 2H ( CH ₂ O), 5.21 (d, 3-6 Hz) 1H (C 6), 5.80 (d, 3-6 Hz) 1H (C 7), 6.91 (d, 3-8 Hz) 1H ( Ar), 7.00 (з) 1H (thiazole), 7.48 (s) 1H (Ar), 7.52 (d, 3-8 Hz) 1H (Ar).

IR (KBr) 3320, 1762, 1680, 1610, 1530.

25.

CS 271 490 82

Mass spectrum: M / z 572 (M-H).

UV spectrum (water): λ _max 260 nm (6,221,800).

HRMS (M ♦ H) for ^C 21 ^H ₁₉ NaN ₅ 0 _g S ₂ :

calculated 572.0522 found 572.0533.

Example 12

Preparation of the monosodium salt (6R-trans) -3- [3,4-bis (acetoxy) benzoyl] oxy] methyl-8-oxo-7 - [(phenoxyacetyl) amino] -5-thia-1-azabicyclo] 4,2, O / oct-2-ene-2-carboxylic acid

Following the procedure and conditions described in Example 10, 129 mg of 1,1-dimethylethyl ester (6R-trans) -3- [3,4-bis (acstoloxy) -benzoyl] oxy / methyl] -8 was reacted. -oxo-7 - [(phenoxyacetyl) amino] -5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid (0.201 mmol) (from Example 9) in 2.5 mL methylene chloride and 220 [mu] l of anisole with 2.2 ml of trifluoroacetic acid, and the reaction mixture is stirred for 6 hours and 30 minutes. After reverse phase chromatography (0-30% acetonitrile in water), 42 mg (34% of theory) of a white powder is obtained.

NMR spectrum (deutriated water):

2.37 (s) 6H (2 OAc), 3.42, 3.70 (dd, 3 to 18 Hz) 2H _(CH2 S), 4.96, 5.16 (dd, 3-12 Hz) 2H (CH ₂ OCO), 5.11 (d, 3 × 6 Hz) 1H (C7), 6.70 (d, 3 × 6 Hz) 1H (C7), 7.01 (d, 3 → 8 Hz) 1H (Ar), 7.05 (m) 1 H (Ar), 7.32-7.43 (g) 4H (Ar), 7.92 (s) 1 H (Ar), 8.00 (d, 3 »8) Hz) 1H (Ar) (CH ₂ OAr overshadowed by HOD).

IR (KBr) 3450, 1782, 1718, 1692, 1608.

Mass spectrum: m / z 607 (M + H).

UV spectrum (water: · Λ _max 236 nm (<£ · 13400).

HRMS (M ♦ H) for C ₂₇ H ₂₄ NaN ₂ 0 ₁₁ S ₂ :

calcd. 607.0900 found 607.1007.

Example 13

Preparation of (6R-) 60 ((7) J (2) H - 7 - [(2-amino-4-thiazolyl) (methoxyimino) acetyl] mono-sodium salt of amino] -3- /// (2 3-dihydroxyphenyl) carbonyl / oxy / methyl-8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid

Using the procedure and conditions described in Example 10, 100 mg (0.118 mmol) of 1,1-dimethylethyl ether (6R- / 6 < X, 7/1 (Z) // - 3 - // (2,3) were reacted. -dihydroxybenzoyl) oxy (methyl) -8-oxo-7 - [(2 -) - (triphanylmethyl) amino] -4-thiazolyl ((methoxyimino) acetyl) amino) -5-thia-1-azabicyclo [4.2], Of O / oct-2-ene-2-carboxylic acid (from Example 6) in 3.0 mL of methylene chloride and 300 µL of anisole with 3.0 mL of trifluoroacetic acid, and the reaction mixture was stirred at room temperature. Stir for 6 hours. Purification by reverse phase chromatography (0 to 30% acetonitrile in water) gave 28.2 mg (42% of theory) of a white powder.

NMR spectrum (deuterated water):

3.30, 3.57 (dd, 3 to 16 Hz) 2H (CHGS), 3.78 (э) 3 H (0CH _3), 4.78, 5.02 (dd, 3-12 Hz), 2H (CH ₂ 0C0). 5.04 (d, 3-6 Hz) 1H (C6), 5.66 (d, 3-6 Hz) 1H (C7), 6.64 (m) 1H (Ar)

6.81 (s) 1H (thiazole), 6.91 (d, 38 Hz) 1H (Ar), 7.24 (d, 36 Hz) 1H (Ar).

IR (KBr): 3350, 1765, 1672, 1612.

Mass spectrum: m / z 572 (M + H).

CS 271 490 82

UV spectrum ^(VOT ^ ^a) ^ _roax ^{245 nm} (£ 21,400 *) .lambda.max 299 nm (£ · 8600)

HRMS (M ♦ H) for ^c ₂₁ H _lg NaN ₅ 0 ₉ S ₂ :

calcd. 572.0522 found 572.0505.

Example 14

Preparation of monosodium salt / 6R- / 6 <*, 7/1 (Z) //-3-//3,4,5-tri(acetyloxy)henzoyl/oxy/methyl/-7-//(2-amino-4- thiazolyl) (methoxyimino) acyl / amino] -8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid

160 mg (0.161 mmol) of 1,1-dimethylethyl ester [6R- (6,7) J (Z)] - (3) - (3,4,5-) was reacted by the procedure and conditions described in Example 10 (0.161 mmol). tris (acetyloxy) benzoyl (oxy) methyl] -7- [(methoxyimino)] - 2 - [(triphenylmethyl) amino] -4-thiazolyl (acetyl) amino] -8-oxo-5-thia-1-azabicyclo] Of 4,2,0 / oct-2-ene-2-carboxylic acid (from Example 7) in 3.2 ml of methylene chloride and 300 µl of anisole with 3.0 ml of trifluoroacetic acid and the reaction mixture is stirred for 6 hours and 30 minutes. minutes. Purification by reverse phase chromatography (0-30% acetonitrile in water) afforded 69 mg (60%) of a white solid.

NMR spectrum (deuterated water):

2.38 (s) 3H (OAc), 2.39 (s) 3H (OAc), 3.51, 3.78 (dd, 3-16 Hz) 2H _(CH2 S), 5.00

5.21 (dd, 3-12 Hz) 2H (CH3 O), 5.24 (d, 3-6 Hz) 1H (C6), 5.82 (d, 3-6 Hz) 1H (C7), 7, 03 (9) 1 H (thiazole), 7.79 (9) 2H (Ar).

IR (KBr) 3360, 1772, 1718, 1675, 1610.

Mass spectrum: m / z 714 (M + H).

UV spectrum (water): _.lambda.max »234 nm (<18,100 f).

HRMS (M + H) for ^C ₂ ^H 25 ^NaN 7 ° 13 5 ^S 2 Calculated 714.0788 Found 714.0745.

Example 15

Preparation of (6R-) 6 <X, 7/3 (Z) // - 7 - [(2-amino-4-thiazolyl) (methoxyimino) acetyl] amino] -3- [2- (3, 2, 3, 3, 4, 3, 4, 3)] 4-Dihydroxyphenyl) acethoxy (methyl) -8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid

175 mg (0.203 mmol) of 1,1-dimethylethyl ester (6R-) 60C, 7D (Z) // - 3 - // 2- (3,4-) were reacted according to the procedure and conditions described in Example 10. dihydroxyphenyl) acetoxy (methyl) -7 - [(methoxyimino)] - 2 - [(triphenylmethyl) amino] -4-thiazolyl] acetyl] amino-8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid (from Example 8) in 3.5 mL of methylene chloride and 350 µL of anisole with 3.5 mL of trifluoroacetic acid and the reaction mixture was stirred for 6 hours. Purification by reverse phase chromatography (0-30% acetonitrile in water) gave 22 mg (19% of theory) of a white solid.

NMR spectrum (deuterated water):

3.27, 3.49 (dd, 3-16 Hz) 2H _(CH2 S), 3.62 (s) 2H (CHgO), 4.00 (s) 3H (0CH _3), 5.00 (d , 3-12 Hz) 1H (1/2 CH ₂ O) (another doublet overshadowed by HOO), 5.17 (d, 3-6 Hz) 1H (C6), 5.82 (d, 3 → 6 Hz) 1H (C7, 6.75 (d, 3-8 Hz) 1H (Ar), 6.86 (9) 1H (Ar), 6.90 (d, 3-8 Hz) 1H (Ar, 7.04 (s) 1H (thiazole).

IR (KBr): 3340, 1763, 167P, 1610.

Mass spectrum: m / z 586 (M-H)

564 (M 4 H of free acid).

27 '

CS 271 490 B2

UV spectrum (water): arm 230 nm (<£ »16 900) arm 260 nm (· · 13 300) ·

HRMS (Μ * H) for C ₂₂ H ₂₁ NaN ₅ 0 _g S ₂ calculated 586.0678 found 586.0664

Example 16

Preparation of (6R-tran8) -7-a (n-1-3 - [(3,4-b8 (acetyloxy) benzoyl) -oxy) methyl] -8-oxo-5-thia-1- (1,1-dimethylethyl) 1,1-dimethylethyl ester monohydrochloride azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid.

A solution of pyridine (1.66 mL, 20.0 mmol) in absolute methylene chloride (60 mL) was cooled to 0 ° C under an argon atmosphere in an ice / water bath and 4.16 g (20.0 mmol) of chloride was added thereto. phosphorous. The mixture was stirred at 0 ° C for 45 minutes. Then, a solution of 8.80 g of (6R-trans) -3- [3,4-b] (acetyloxy) -benzoyl (oxy) methyl] -8-oxo-7- (1,1-dimethylethyl ester) is added dropwise to this solution. (phenoxyacetyl) amino] -5-thia-1-azabicyclo [4.2.0-oct-2-ene-2-carboxylic acid (from Example 12) in 40 mL of absolute methylene chloride over 20 minutes. The reaction mixture was stirred at 0 ° C for 1 hour and 30 minutes and then 24 ml of 1-propanol was added dropwise. Stirring is continued for 1 hour and then 60 ml of water are added and stirring is continued for 40 minutes. The volatiles were removed by evaporation under reduced pressure. To the remaining aqueous solution was added 300 ml of diethyl ether with vigorous stirring to precipitate the product as a white crystalline solid. Yield 5.28 g (71% of theory).

NMR Spectrum (perdeutrogenated dimethylsulfoxide):

1.49 (s) 9 H (χ-bu), 2.32 (s) 3 H (OAc), 2.33 (s) 3 H (OAc), 3.78, 3.87 (dd, 3-18 Hz) 2H _(CH2 S), 4.97 (m) 1H (C6), 5.24 (m) ЗН (C7, CHgO), 7.47 (d, 3-8 Hz) 1H (Ar);

7.85 (s) 1H (Ar), 7.92 (d, 3-8 Hz) 1H (Ar), 9.05 (broad) 3H (NH ₃ ⁺ ).

IR (KBr) 3400, 1778, 1772.

UV spectrum (ethanol): λ _max 238 nm (f 1580).

Example 17

Preparation of 1,1-dimethylethyl ester (6R-) - [6 ', 7' (Z)] - 3 - [3,4-bis (acetvloxy) benzoyl / oxy] methyl-7 - [(2-amino- 4-thiazolyl) [1,1-dimethyl-2- (1,1-dimethylethoxy) -2-oxoethoxy] imino (acetyl) amino-8-oxo-5-thia-1-azabicyclo [4.2.0] oct -2-ene-2-carboxylic acid

A suspension of 1.00 g (1.84 mmol) of 1,1-dimethylethyl ester (6R-trans) -7-amino-3 - [(3,4-bus (acstyloxy) benzoyl) oxymethyl] -8-oxo monohydrochloride -5-thia-1-azabicyclo [4.2.0-oct-2-ene-2-carboxylic acid (from Example 16) in 25 mL of methylene chloride was thoroughly washed with 25 mL of a 4: 1 solution of brine. The organic solution was separated and dried over magnesium sulfate. 1.10 g (2.30 mmol) of S-2-benzothiazolyl-2-amino-[(Z) - [1- (tert-butoxycarbonyl) -1-methylenethoxy] imino] thio-4 are added in one portion to this solution. -thiazoleacetate. The resulting solution was stirred for 16 hours and the solvent was removed under reduced pressure. The residue was chromatographed on silica gel using n-hexane / ethyl acetate (1: 1) then ethyl acetate alone to give 1.16 g (77%) of a cream colored solid.

NMR spectrum (dsuterochloroform):

1.41 (s) 9H (t-bu), 1.52 (s) 9H (tbu), 1.56 (β) 3H (CH ₃ ), 1.58 (s) 3H (CH ₃ ),

2.29 (s) 6H (20 Ac), 3.38, 3.59 (dd, 3-18 Hz) 2H (CH 3 S), 5.04 (m) 2H (C 6, 1/2 CH ₂ O)

5.38 (d, 3-12 Hz) 1H (1/2 CHgO), 6.01 (dd, 3-6 Hz, 3-10 Hz) 1H (C7), 6.10 (s)

2H (NH _2), 6.88 (s) 1H (thiazole), 7.25 (d, 3-12 Hz) 1H (NH), 7.68 (d, 3-8 Hz) 1H (Ar), 7 82 (s) 1H (Ar) 7.91 (d, 3-8 Hz) 1H (Ar).

CS 271 490 B2

-) ъ о

IR (KBr): 3300, 1782, 1722, 1688, 1535.

Mass epaction: m / z 818 (M-H).

UV spectrum (ethanol): λ _max 235 nm (S 28 28 450).

Example 18

Preparation of (6R- / 6S, 7D (Z)) - 3 - /// 3,4-bis (acetyloxy) benzoyl / oxy / methyl / -7 - //// 1,1-dimethylethyl ester (2- amino-2-oxoethoxy) imino [4-thiazolyl] acetyl / amino] -8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid

A suspension of 234 mg (0.430 mmol) of 1,1-dimethylathylaether (6R-tans) -7-amino-3 - [(3,4-bis (acetyloxy) benzoyl) oxy] methyl] -8-oxo-5-thia monohydrochloride -1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid in 10 mL of methylene chloride was washed with 10 mL of a 4: 1 mixture of brine and saturated sodium bicarbonate. The organic solution was separated and dried over magnesium sulfate. . This solution was added dropwise to a solution of S- (2-benzothiazolyl) -Z-aminothi-4-thiazole-glyoxylate O- (carbamoylmethyl) oxime (157 mg, 0.400 mmol) in 5 ml of methylene chloride and 8 ml of absolute dimethylformamide. Stirring was continued for 3 hours and the solvents were removed under reduced pressure. The residue is chromatographed using methylene chloride containing 0-10% methanol as eluent to give 220 mg (70% of theory) of a cream colored product.

NMR Spectrum (CDCl3):

1.51 (β) 9 H (t-bu), 2.28 (s) 3 H (OAc), 2.29 (β) 3 H (OAc), 3.48, 3.51 (dd, □ 18 Hz)

2H (CH ₂ S), 4.60, 4.81 (dd, □ 18 Hz) 2H (CH ₂ CO ₂ ), 4.96, 5.37 (dd, □ - 12 Hz) 2H (CH ₂ O) , 5.10 (d, □ π 6 Hz) 1H (C6), 5.83 (br) 2H _(NH2), 5.83 (br) 2H _(NH2). 6.01 (dd, □ 6 Hz, 3> 10 Hz) 1H (C7), 6.76 (s) 1H (thiazole), 7.27 (d, □ - 8 Hz)

1 H (Ar), 7.85 (s) 1 H (Ar), 7.93 (d, □ 8 Hz) 1 H (Ar).

IR (KBr) 3450, 3350, 1778, 1720, 1680, 1532.

Mass spectrum: m / z 733 (M + H).

UV spectrum (ethanol): λ _max 236 nm (£ 28,700).

Example 19

Preparation of biododiol (6R-) 6Z, 7Z- (Z) - (3) - [3,4-b] - (acetyloxy) benzoyl (oxy) methyl] -7 - [(2-amino-4-thiazolyl)] (1-Carboxy-1-methylethoxy) amino (acetyl) amino] -8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid

Following the procedure and conditions described in Example 10, it was reacted

247 mg (0.301 mmol) of 1,1-dimathylethyl ester (6R- / 60o (1.7 / 1 (Z)) - 3 - [] 3,4-bis (acetyloxy) benzoyl (oxy) matyl] -7-] // (2-amino-4-thiazolyl) / 1,1-dimethyl-2- (1,1-dimethylethoxy) -2-oxoethoxy / imino / acetyl / amino / -8-oxo-5-thia-1-azabicyclo Of (4,2,0) oct-2-ene-2-carboxylic acid (from Example 17) in 2.5 ml of methylene chloride and 250 µl of anisole with 2.5 ml of trifluoroacetic acid and the reaction mixture was stirred for 7 hours. After purification by reverse phase chromatography (0 to 30% acetonitrile in water), 61.2 mg (27% of theory) of a white solid is obtained.

NMR spectrum (pardeuterated dimethylsulfoxide):

1.30 (s) 3H ([delta] 1.37 (s) 3H (CMe ₂ ), 2.20 (s) 3H OAc), 2.21 (s) 3H (OAc), 3.49 (d, □ · 18 Hz) 1H (1/2 CH ₂ S, second half shaded by HOD), 4.87, 5.23 (dd, □ 12 Hz) 2H (CH ₂ O), 4.96 (d, □ ») 6 Hz) 1H (C 6), 5.60 (dd, □ 6 Hz, □ 10 Hz) 1H (07), 6.66 (s) 1H (thiazole), 7.11 (broad) 3H (NH, NH ₂ ), 7.37 (d, 3 → 8 Hz) 1 H (Ar), 7.78 (s) 1 H (Ar), 7.85 (d, □ 8 Hz) 1 H (Ar).

IR (KBr) 3400, 1770, 1718, 1672, 1608.

CS 271 490 B2

2 £

Mass spectrum: m / z 650 (M + H).

UV spectrum (ethanol): λ 235 nm (<£ 20,000).

HRMS (M + H) calcd for C ₂₈ H ₂₅ NaN ₅ 0 ₁₃ S ₂ s 772.0584 found 772.0543

Example 20

Preparation of the disodium salt of (6R-) 6 (X, 7) J (Z) H-7 - [(2-amino-4-thiazolyl)] - 1-carboxy-1-methylethoxy (1-amino) acetyl / amino] -3- [(3,4-dihydroxybenzoyl) oxy] methyl] -8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-sn-2-carboxylic acid

A solution of 220 ml (0.227 mmol) of 1,1-dimethylethyl ester (6R- / 6,6,7,7 (Z)) - 3 - [] 3,4-bis (acetyloxy) -benzoyl (oxy) methyl] - 7 - [(amino-4-thiazolyl)] - 1,1-dimethyl-2- (1,1-dimethylethoxy) -2-oxoethoxy (imino) acetyl] amino] -8-oxo-5-thia-1 The azabicyclo [4.2.0-oct-2-ene-2-carboxylic acid in 10 mL of methylene chloride and 1 mL of anisole was stirred at 0 ° C and 10 mL of trifluoroacetic acid was added with stirring. Stirring was continued for 6 hours at 0 ° C and then the volatiles were removed under reduced pressure. 5 ml of ethyl acetate, 5 ml of saturated sodium bicarbonate solution and 5 ml of methanol are added to the oily residue and the biphasic mixture is stirred for 30 minutes. The solution is concentrated under reduced pressure to a volume of about 5 ml and then 5 ml of ethyl acetate and 5 ml of saturated sodium bicarbonate solution are added thereto. The resulting solution is stirred for 15 minutes. The aqueous layer was separated to give 60 mg (33% of theory) of a white solid after purification by reverse phase chromatography (0-20% acetonitrile in water).

NMR spectrum (perdeuterated dimethylsulfoxide):

1.40 (s) 3H (CMe ₂ ), 1.49 (s) 3H (CMe ₂ ), 3.31, 3.49 (dd, □ 16 Hz) 2H (CH ₂ S), 4.96, 5.04 (dd, □ - 12 Hz) 2H (CH 3 O), 5.05 (d, □ 6 Hz) 1H (C 6), 5.68 (m) 1H (C 7), 6.73 (m) 2H (Ar, thiazole), 7.18 (br) 2H _(NH2), 7.27 (d, □ «8 Hz) 1H (Ar), 7.34 (s) 1H (Ar), 11.70 (d 0-10 Hz) 1 H (NH).

IR (KBr): 3350, 1762, 1670, 1598.

Mass spectrum: m / z 666 (M 2 H).

UV spectrum (ethanol) ´ 218 nm (<£ »25 050)‘

Amax ^{262 (} <6 ^{'17 60} °) ·

HRMS (M * H) calcd for C ₂₄ ^H 21 ^N 5 ^O 11 ^S 2 666.0553 found 666.0619.

Example 21

Preparation of [6R- / 60o (1,7fl (S) ^x ) -1-dimethylethyl ester] - 3 - [(3,4-bis (acetyloxy) benzoyl) oxy] methyl] -7- H - ( 4-ethyl-2,3-dioxo-1-piperazinyl) carbonyl (amino) -phenylacetyl] amino-8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid acid

Suspension 275 mg (0.506 mmol) of (6R-trans) -7-amino-3 - [(3,4-b8 (acetyloxy) benzoyl) oxy / methyl] -8-oxo-5-thia 1,1-dimethylethyl ester monohydrochloride The 1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid in 10 mL of methylene chloride was washed with 10 mL of a 4: 1 mixture of brine and saturated sodium bicarbonate. The organic solution was separated and dried over magnesium sulfate. This solution was added to an activated ester solution (prepared by adding 69 mg (0.506 mmol) of N-hydroxybenzotriazole) followed by 104 mg (0.506 mmol) of 1,3-dicyclohexylcarbodiimide to a solution of 161 mg (0.506 mmol) of <X - // (4-ethyl) -2,3-dioxo-1-peperazinyl) carbonyl / amino / phenylacetic acid in 10 ml of absolute tetrahydrofuran and

CS 271 490 82 for 45 minutes). The solution was stirred for 16 hours, filtered and the solvent was removed under reduced pressure. The residue was chromatographed using ethyl acetate as eluent to give 250 mg (61%) of a cream colored solid.

NMR Spectrum (CDCl3):

1.20 (t, J = 3 Hz) 3H (Et), 1.52 (s) 9H, (δ -bu), 2.32 (β) 6H (2OAc), 3.2-3.7 (m) 6H (CH ₂ S, Et, 1 / 2NCH ₂ CH ₂ N), 3.93 (m) 1H (1 / 4NCH ₂ CH ₂ N), 4.11 (m) 1H (1 / 4NCH ₂ CH ₂ N), 4.91 (d, 3 »6 Hz) 1H (C6), 5.00, 5.30 (dd, 3-14 Hz), 2H (CH ₂ 0), 5.58 (d, 3 · 8 Hz) 1H (NCHCPh), 5.84 (dd, 3-6 Hz, 3-10 Hz) 1H (C7), 6.83 (d, 3-10 Hz) 1H (NH),

7.20-7.35 (< n) 6H (Ar), 7.80 (s) 1H (Ar), 7.89 (d, 3-8 Hz) 1H (Ar), 10.04 (d, 3-8) Hz) 1 H (nH).

IR (KBr): 3320, 1780, 1718, 1688.

UV spectrum (ethanol): arm 225 nm (δ = 23,300), arm 270 cm (»» 10,900).

Example 22

Preparation of (6R-) 60C, 7β (S) ^x H -3- (3) 3,4-bis (acetyloxy) benzoyl / oxy / methyl-7 H- (4-ethyl) -2-sodium salt 3-dioxo-1-piperazinyl) carbonyl (amino) phenylacetyl / -amino) -8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid

Following the procedure and conditions as described in Example 10 is reacted with 100 mg (0.124 mmol) of 1 J-dimethylethyl / 6R- / 6 <d, 7 / $ (S) ^x // - 3 - /// 3,4-bis (acetyloxy) benzoyl (oxy) methyl] -7- [alpha] - [(4-ethyl-2,3-dioxo-1-piperazinyl) carbonyl / amino] phenylacetyl] amino] -8- oxo-5-thia-1-azabicyclo [4.2.0] oct-2-an-2-carboxylic acid (from Example 21) in 5 ml of methylene chloride and 500 µl of anisole with 5 ml of trifluoroacetic acid, the reaction being The mixture was stirred for 5 hours and purified by reverse phase chromatography (0-30% acetonitrile in water) to give 48 mg (50%) of a white solid.

NMR spectrum (perdeuterated dimethylsulfoxide):

1.08 (t, 3 → 8 Hz) 3H (NEt), 2.31 (s) 6H (20Ac), 3.21-3.60 (m) 6H (CH ₂ S, NEt, 1/2 NCH ₂ CH ₂ N), 3.91 (cn) 2H (1/2 NCH ₂ CH ₂ N), 4.92 (d, 3-6 Hz) 1H (C 6), 4.95, 5.25 (d, 3 12 Hz) 2H (CH ₂ O), 5.57 (m) 1 H (C 7), 5.75 (d, 3 → 8 Hz) 1 H (NCHCPh), 7.26 7.48 (m) 6 H (Ar) ), 7.82 (s) 1H (Ar), 7.89 (d, 3-8 Hz) 1H (Ar), 9.41 (d, 3-8 Hz) 1H (NH) 9.86 (d, 3-10 Hz 1H (NH).

IR (KBr) 3420, 3305, 1772, 1715, 1682, 1610.

UV spectrum (ethanol): λ _max ²³⁰ nm (δ »22,600).

HRMS (M * H) calcd for ^C 34 ^H 33 ^NaN 5 ° 13 ^S 774.1693 found 774.1729.

Example 23

Preparation of the monosodium salt of (6R-6 ', 7y3 (S) ^x H - 7 - H? X - // (4-ethyl-2,3-dioxo-1-plperazinyl) carbonyl / amino) -phenyl acid 1- (amino) -3 - [(3,4-dihydroxy) benzoyl] oxy] methyl-8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-en-2 -carboxylic acids

According to the procedure and conditions described in Example 20, 150 mg (0.186 mmol) of 1,1-dimethylethyl ester (6R- / 60 °, 17/1 (S) ^x H-3 - /// 3) were reacted. 4-bis (acetyloxy) benzoyl (oxy) methyl 1- (1H) - (4-ethyl-2,3,3-dioxo-1-piperazinyl) carbonyl (amino) -phenylacetyl (amino) - 8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid (from Example 21) in 5 mL of methylene chloride and 500 µL of anisole with 5 mL of trifluoroacetic acid. After completion of the reaction, the mixture was stirred with a mixture of ethyl acetate and saturated aqueous bicarbonate solution

CS 271 490 B2 of sodium citrate and methanol in a 1: 1: 1 ratio. After chromatographic purification using reverse phase (0-20% acetonitrile in water), 68 mg (53% of theory) of a white solid are obtained.

NMR spectrum (perdeuterated dimethyl sulfoxide):

1.01 (t, □ - 8 Hz) 3H (Net), 3.14, 3.39 (dd, □ 18 Hz) 2H (CH ₂ S), 3.30 - 3.60 (m) 4H ( NEt), 1/2 NCH ₂ CH ₂ N), 3.85 (m) 2H (1/2 NCH ₂ H ₂ N), 4.80. 5.08 (dd, □ 12 Hz) 2H (CH ₂ O), 4.83 (d, □ - 6 Hz) 1H (C 6), 5.48 (m) 1H (C7), 5.61 (d Δ → 8 Hz) 1H (NCHCPh), 6.69 (d, δ → 8 Hz) 1H (Ar), 7.16-7.41 (m) 7 H (Ar), 9.33 (broad) 1H ( NH), 9.78 (d, □ 8 Hz) 1H (NH).

IR (KBr) 3400, 3300, 1762, 1712, 1680, 1603.

Mass spectrum: m / z 690 (Μ + H).

UV spectrum (ethanol): _λmax 263 nm (<£ 17,900).

HRMS (Μ + H) for ° 3ο ^Η 29 ^Ν 5 ° 11θ ^Νβ calculated 690.1482 found 690.1517

Example 24

Preparation of (6R-) 6 *, 7 (R) -3- [3,4-bis (acetyloxy) benzoyl] oxymethyl] -7- (2-amino-4-) monosodium salt thiazolyl // (2-amino-2-oxoethoxy) imino (acetyl) amino] -8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid

Following the procedure and under the conditions described in Example 10, 200 mg (0.272 mmol) of (6R-) - (6) - (1) -dimethyl-ethyl ester 1,1-dimethylethyl ester was reacted (3) - (3) - (3) 4-bie (acetyloxy) benzoyl (oxy) methyl] -7- [2-amino-4-thiazolyl] (2-amino-2-oxoethoxy) imino (acetyl) amino) -8-oxo-5- of thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid (from Example 18) in 4 ml of methylene chloride and 400 µl of anisole 8 with 4 ml of trifluoroacetic acid while stirring for 6 hours. hours. Purification by reverse phase chromatography (0-30% acetonitrile in water) gave 40 mg (21% of theory) of a white solid.

NMR spectrum (perduterated dimethylsulfoxide):

2.28 (β) 3H (OAc), 2.29 (s) 3H (OAc), 3.57 (d, 18 18 Hz) 1H (1/2 CH ₂ S, 1/2 CH ₂ S overshadowed by HOD ), 3.99 (β) 2H (CH ₂ O), 4.94, 5.27 (dd, □ 12 12 Η) 2H (CH ₂ OC), 5.01 (d, □ 6 Hz) 1H (C6 ), 5.60 (dd, □ «6Hz, □« 10 Hz) 1H (C7), 6.84 (s) 1H (thiazole), 7.10 (s) 2H _(NH2), 7.28 2H (NH 3, 7.11 (d, □ 8 Hz) 1H (Ar), 7.84 (e) 1 H (Ar), 7.89 (d, □ 8 Hz) 1H (Ar).

IR (KBr): 3440, 1768, 1710, 1710, 1678, 1610.

Mass spectrum: m / z 699 (M + H).

UV spectrum (water); _λmax ²³⁴ nm 26,260).

HRMS (M + H) calcd for ^C 26 ^H 26 ^NaN 6 ° 12 ^S 2 699.0791 found 699.0822.

Example 25

Preparation of (6R-cis) -3 - [(aminocarbonyl) oxy] methyl (-7-methoxy-8-oxo-7- (2-thienylacetyl) amino-5-thia-1-azabicyclo) (4-nitrophenyl) methyl ester 4,2,0 / oct-2-ene-2-carboxylic acid

3,7 3.75 g (8.30 mmol) of p-nitrobenzyl bromide in 100 ml of absolute dimethylformamide

CS 271 490 B2 add 3.75 g (8.30 mmol) of (6R-cis) -3 - [(aminocarbonyl) oxy / methyl] -7-methoxy-α-oxo-7 - / (2- thlenylacetyl) amino] -5- thia-1-azabicyclo [4.2.1.0] oct-2-ene-2-carboxylic acid. The mixture was stirred for 2 hours 30 minutes and then 100 ml of ethyl acetate was added thereto. The resulting solution was washed with brine, washed with magnesium sulfate, and the solvent was removed under reduced pressure. The residue was chromatographed on silica gel using 1: 1 ethyl acetate: n-hexane and then 2: 1 as the eluent. The truncated product ae was crystallized from an acetate / n-hexane mixture to yield 3.4 g (72% of theory) of the product.

NMR spectrum (deutorochloroform):

3.31, 3.49 (dd, 3-18 Hz) 2H (CH 3 S), 3.44 (a) 3 H (OCH ₃ ), 3.79 (x) 2H (CH 3 CON), 4.67 (broad) 2H (NH _2), 4.82, 5.12 (dd, 3-14 Hz), 2H (CH> 0), 5.03 (s) 1H (C6), 5.31, 5.60 (dd, 0 -14 Hz) 2H (CH3), 6.44 (s) 1H (NH), 6.99 (m) 2H (thiophene), 7.25 (m) 1H (thiophene), 7.56 (d, 3-8 Hz) 2H (Ar) 8.19 (d, 3-8 Hz) 2H (Ar).

IR (KBr) 3465, 3335, 1785, 1728, 1698, 1522, 1348 ·

Example 26

Preparation of (6R) -3- (Iodomethyl) -7-methoxy-8-oxo-7 - [(2-thienylacetyl) amino] -5-thia-1-azabicyclo [4.2.0] (4-nitrophenyl) methyl ester oct-2-ene-2-carboxylic acid

(3.0 g (5.33 mmol) of (6R-cis) -3 - [(aminocarbonyl) oxy] methyl) -7-methoxy-8-oxo-7 - [(2-thienylacetyl) (4-nitrophenyl) methyl ester] amino) -5-thla-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid (from Example 25) in 75 mL of methylene chloride and 1.40 mL (10.3 mL) at room temperature mmol) trimethylsilyl iodide. The reaction mixture was stirred for 30 minutes and then washed with aqueous sodium hydrogencarbonate solution and dried over magnesium sulfate. reagents. 2.88 g (86% of theory) of a pale yellow solid are obtained.

NMR Spectrum (CDCl3):

3.45, 3.60 (dd, 3-16 Hz) 2H (CH 3 S), 3.49 (s) 3 H (OCH ₃ ), 3.94 (β) 2H (CH ₂ CO 3),

4.36, 4.50 (dd, 3-8 Hz) 2H _(CH2 3), 5.07 (s) 1H (C6), 5.36, 5.46 (dd, 3-16 Hz) 2H ( CHgAr), 8.46 (β) 1H (NH), 7.05 (m) 2H (thiophene), 7.30 (m) 1H (thiophene), 7.65 (d, 3 · 8 Hz) 2H (Ar 8.27 (d, 3-8 Hz) 2H (Ar).

Example 27

Preparation of (6R-cis) -3- [3,4-bia (acetyloxy) benzoyl] oxy (methyl) -7-methoxy-8 * oxo-7 - [(2-thienylacetyl) amino] - (4-nitrophenyl) methyl ether 5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acids

(1.90 g (3.02 mmol) of (6R-cis) -3- (iodomethyl) -7-methoxy-8-oxo-7 - [(2-thienylacatyl) amino) -5-nitrophenyl) methyl] -5- of thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid (from Example 26) in 15 ml of absolute dimethylformamide at 0 ° C is added 0.78 g (3.00 mmol) in one portion. 3,4-diecetoxybenzoic acid sodium salt · The reaction solution is stirred for 30 minutes and then 100 ml of ethyl acetate are added thereto. The resulting solution is washed with brine (3 x 50 ml), dried and magnesium sulphate and the solvent is evaporated under reduced pressure. . Chromatographic purification using 1: 1 n-hexane: ethyl acetate as eluent gave 1.30 g (58%) of a mixture of 42 α-α-isomers. This product was dissolved in 30 ml of methylene chloride at 0 ° C and 400 mg (2.32 mmol) of m-chloroperbenzoic acid was added in one portion. The mixture was stirred for 2 hours at 0 ° C, then washed with an aqueous solution of sodium bicarbonate, dried over magnesium sulfate, and the solvent removed under reduced pressure. Crystallization from ethyl acetate / n-hexane (5: 2) yields 870 mg (66%) of the sulfoxide in

CS 271 490 B2 to form a single leomer. The mother liquor was chromatographed on silica gel with ethyl acetate / methylene chloride (1: 1) to give 320 mg (24%) of the isomeric sulfoxide. The combined sulfoxides (1.19 g, i.e. 90% of theory) 9Θ are reduced by dissolving in 5 mL of absolute dimethylformamide at -20 ° C and adding 0.34 mL of phosphorous tribromide followed by stirring for 1 hour. Removal of the solvent under reduced pressure followed by chromatography on silica gel using ethyl acetate / n-hexane (1: 1 ee) afforded 967 mg (83%) of the title compound.

NMR Spectrum (CDCl3):

2.29 (β) 6H (20AC), 3.33, 3.64 (dd, 0 to 18 Hz) 2H _(CH2 S), 3.44 (s) 3H (0CH _3), 3.08 (β ) (CH ₂ CO 3), 4.98 (d, □ 12 Hz) 1H (1/2 CH ₂ O), 6.04 (a) 1H (C 6), 5.34 (m) 3 H (1/2 CH ₂ O, CH ₂ ), 6.40 (s) 1H (NH), 6.99 (m) 2H (thiophene), 7.24 (m) 2H (thiophene, Ar), 7.64 (d, □) 8 Η) 1 H (Ar), 7.78 (β) 1 H (Ar), 0.07 (d, □ 0 Hz) 1 H (Ar), 0.16 (d, □ 8 Hz) 1 H (Ar).

MS (KBr): 3325, 1775, 1722, 1700, 1520, 1348.

Mass spectrum: m / z 740 (M + H).

Example 28

Preparation of (6R-cis) -3 - [(3,4-bis (acetyloxy) benzoyl) oxy] methyl-7-methoxy-8-oxo-7 - [(2-thlenylacetyl) amino] -5-thia] monosodium salt 1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid

To a solution of (6R-C 18) -3- [3,4-bis (acetyloxy) benzoyl] oxy (methyl) -7-methoxy-8-oxo-7- (4-nitrophenyl) methyl ether (4-nitrophenyl) methyl (2-Thienylacetyl) amino] -6-thia-1-azabicyclo] oct-2-ene-2-carboxylic acid in 5 mL of tetrahydrofuran was added 100 mg of 5% platinum on carbon. The mixture is hydrogenated at 50 psi for 48 hours. To remove catalyst ee, the mixture was filtered and the solvent was evaporated under reduced pressure. The residue is taken up in 3 ml of ethyl acetate and 3 ml of a 2% aqueous solution of sodium bicarbonate and the solution is stirred vigorously for 15 minutes. The aqueous layer was separated and chromatographed (reverse phase, 0-30% acetonitrile in water). After freeze drying of the appropriate fractions, 45 mg (53% of theory) of a white solid are obtained.

NMR spectrum (perdeuterated dimethylsulfoxide):

2.32 (s) 6H (20AC), 3.19, 3.59 (dd, □ · 18 Hz) 2H _(CH2 S), 3.49 (s) 3H _(OCH3) 3.79;

3.89 (dd, D 16 Hz) 2H (CH ₂ CO), 4.98, 5.17 (dd, □ 14 Hz) 2H (CH ₂ O), 5.00 (а) 1H (C6), 6.96 (m) 2H (thuophene), 7.38 (m) 1H (thiophene), 7.45 (d, □ - 8 Hz) 1H (Ar), 8.84 (s) 1H (Ar), 8 91 (d, □ 8 Hz) 1 H (Ar), 9.41 (m) 1H (NH).

IR (KBr) 3400, 1770, 1618, 1612.

Mass spectrum: m / z 872 (M + H).

UV spectrum (ethanol): Λ _mex 236 nm (<^ · 16 620)

HRMS (m, 4 H) for C ₂₄ H _2S NaN _{2 0] L1} S2 calcd 627.0719 found 627.0676.

CS 271 490 82

Example 29

Preparation of mono-sodium (6R-cio) -3 - [(3,4-dihydroxybenzoyl) oxy] methyl] -7-mothoxy-O-oxo-7 - [(1 H snylaoatyl) amino] -α-thia-1-azabicyclo (4,2,0) oct-2-sn-2-carboxylic acid

65 mg (0.104 mmol) of (6R-cie) -3- [3,4-bie (acetyloxy) banzoyl] oxy / methyl] -7-methoxy-B-oxo-7 - / (2- thianylacetyl) amino] -5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid (from Example 28) in 4 ml of saturated aqueous sodium bicarbonate solution is added 1 ml of methanol. The reaction solution was stirred at room temperature for 1 hour. Methanol ee was removed under reduced pressure and the remaining aqueous solution was chromatographed (reverse phase, 0-20% acetonitrile in water). Lyophilization of the appropriate fractions gave 52 mg (92%) of a white solid.

NMR spectrum (perdeuterated dimethylsulfoxide):

3.11 (dd, 3-18 Hz) 1H (1/2 CH ₂ S, second half overshadowed by HOD), 3.37 (s) 3H (OCH ₃ ), 3.78 (dd, 3 »16 Hz) 2H (CH ₂ CO), 4.83, 5.01 (dd, 3 → 12 Hz, 2h (CH ₂ O)), 4.98 (s) 1H (C 6), 5.40 (broad) ЗН (20H NH 6.38, 6.95 (m) 1H (Ar), (m) 2H (thiophene), 7.11 (s) 1H (Ar), d, 3-8 Hz) 1H (Ar), 7, 36 (m) 1H (thiophene).

IR (KBr): 1762, 1675, 1610.

Mass spectrum: m / z 543 (M + H). '

UV spectrum (ethanol): 222 _max 222 nm (£ 15 15,960)

Λ max ^{255 nm č 10 280} >

HRMS (M ♦ H) for ^C ₂ ^H 20 ^N 2 2 ° 9 ^S 2 ^Na: calculated 543.0508 found 543.0506

Example 30

Preparation of (6R-6 ', 7-Dimethyl-ethyl ester) (f) (Z) 3 - [3 - [(3,4-b8 (acetyloxy) benzoyl) thio] methyl] -8-oxo-7 - (2) - ((triphenylmethyl) amino) -4-thiazolyl] (methoxyimino) acetyl / amino] -5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid

Following the procedure and conditions described in Example 4, 61 mg (0.220 mmol) of 3,4-bis- (acatyloxy) banzancarbothioic acid monosodium salt in 2 ml of absolute dimethylformamide was added to 171 mg (0.208 mmol) of 1,1-dimethylethyl ether. (6R- / 6 (X, 7 (J)) -3- (iodomethyl) -7- [2- (methoxyimino) -2- [2- (tritylamino) -4-thiazolyl] acetyl / amino] -8 * oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid in 2 ml of dimethylformamide, the reaction mixture is stirred for 3 hours and chromatographed on silica gel using methylene chloride; methanol (9: 1) gave a cream solid (112 mg, 57%).

NMR Spectrum (CDCl3):

1.54 (s) 9H (t-Bu), 2.31 (s) 6H (2 OAc), 2.32, 2.62 (dd, 0 to 18 Hz) 2H _(CH2 S), 4.05 , 4.30 (dd, 0-10 Hz) 2H (CH3SCO), 5.02 (d, 3.6 Hz) 1H (C6), 5.90 (dd, 36 Hz, 3 → 10 Hz) 1H ( C7), 6.68 (d, 3-10 Hz) 1H (NH), 6.70 (s) 1H (thiazole), 6.99 (broad) 1H (NH), 7.26 (s) 16H ( CPh ₃ , Ar), 7.81 (α) 1H (Ar), 7.86 (d, 3-8 Hz) 1H (Ar).

IR (KBr): 178, 1715, 1665.

UV spectrum (ethanol): λ _max 214 nm (£ 36,000)

Арах ²⁶⁷ ™ ^{29 750} >

Mass spectrum: m / z 948 (M + H).

CS 271 490 B2

Example 31

Preparation of mono-sodium [6R- / 6 *, 7/1 (Z)] - 7 - [(2-amino-4-thiazazolyl) (m-methoxyimino) acetyl] amino] -3- /// (3,4- bis- (acetyloxy) benzoyl (thio) methyl] -8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid

Following the procedure and under the conditions described in Example 10, 208 mg (0.219 mmol) of 1,1-dimethylethyl 6R- / 6 (X), 74 (Z) // - 3 - // (3,4) was reacted. -bis (acetyloxy) benzoyl) thio (methyl) -B-oxo-7 - [(2 -) - (triphenylmethyl) amino] -4-thiazolyl] (methoxyimino) acetyl (amino) -5-thia-1-azabicyclo] 4.2, O / oct-2-ene-2-carboxylic acid (Example 30) in 4 ml of methylene chloride and 500 µl of anisole with 4 ml of trifluoroacetic acid, the reaction mixture is stirred for 5 hours and then worked up as described in Examples 31, After purification by 8 reverse phase chromatography (0 to 20% acetonitrile in water) to give 77 mg (52% of theory) of a white powder.

NMR spectrum (deuterated water):

2.40 (E) SH (2 OAc), 3.39, 3.73 (dd, IQ 3 Hz), 2H (CHGS), 01 (s), 3 H (OCH _g), 3.96,

4.39 (dd, 3 · 14 Hz) 2H (CH 3SCO), 5.21 (d, 3 · 6 Hz) 1H (C 6), 5.81 (d, 3 → 6 Hz) 1H (C7), 7, 03 (α) 1H (thiazole), 7.45 (d, 38 Hz) 1H (Ar), 7.89 (s) 1H (Ar), 8.98 (s) 1H (Ar), 8.98 (s) а) 1H (Ar) ·

UV 8pectrum (water): A _max 237 nm ((20,200)

Лжах ²⁷⁵ · ^{17 00} °)

Mass spectrum: m / z 672 (M + H). .

Example 32

Preparation of [6R- (60C, 7R) (Z)] - 3 - [(3,4-bie (acetyloxy) -2,5-dichlorobenzoyl) oxy] methyl] -7- (1,1-dimethylethyl ether) ((methoxyimino)) ((triphenylmethyl) amino) -4-thiazolyl (acstyl) amino] -8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-an-2-carboxylic acid

Following the procedure and under the conditions described in aa in Example 4, 87 mg (0.264 mmol) of 3,4-bls (aoetyloxy) -2, β-di-chlorobenzoic acid sodium salt in 2 ml of dimethylformamide and 208 mg (0.253 mmol) were reacted. (6R- / 6 *, 7R (Z)) - 3- (iodomethyl) -7- [2- (methoxyimino) -2- [2- (tritylamino) -4-thiazolyl] 1,1-dimathylothyleator] -aoethyl / amino / 8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid in 2 ml of dimethylformamide, the reaction mixture is stirred for 4 hours and after chromatographic purification on the column of melcogol of use, the malhyianohler was bleached with methanol in a ratio of U to 1 as the reagent and a 1 / n mg (/> theory) of the title compound.

NMR Spectrum (CDCl3) m.p.

1.50 (α) 9H (t-bu), 2.36 (α) 6H (2 OAc), 2.36, 2.60 (dd, 3-18 Hz) 2H (CH 3 S), 4.03 (α) ) 3 H (0CH _3), 5.01 (m) 2H (1/2 CHgO, C6), 5.38 (d, 3 »14 Hz) 1H (1/2 CHgO), 5.92 (dd, 3 · 6 Hz, 3 10 Hz) 1H (C7), 6.68 (m) 2H (NH, thiazole), 7.12 (β) 15H (CPh ₃ ), 7.81 (а) 1H (Ar) ·

Example 33

Preparation of the mono-sodium salt of (6R-) 60,7,7 (J) - (7) - (2-amino-4-thiazolyl) (m-methoxyimino) acetyl / amino] -3 - /// (3,4-bi8) (acetyloxy) -2,5-dichlorobenzoyl) (oxy) methyl] -8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid

A solution of 146 mg (0.146 mmol) of 1,1-dimethylethyl ether (6R-) 6,7,7 (J) - (3) - (3,4-b18 (acG-thyloxy) -2,5-dichlorobenzoyl) oxy [methyl] -7 - [(methoxyimino)] - 2 - [(triphenylmethyl) amino-4-thiazolyl] amino] -8-oxo-5-thia-1-azabioyklo [4.2.0] oct-2-ene Of 2-carboxylic acid (Example 32) in 4.6 ml of 70% aqueous formic acid solution was stirred at room temperature for 3 hours

CS 271 490 82 rooms. The solvent was removed under reduced pressure and the residue was chromatographed on silica gel (eluting with methylene chloride containing 0 to 100% ethyl acetate) to give 80.7 mg (72% of theory) of the detritylated product. A solution of this product (39 mg, 0.051 mmol) in methylene chloride (1.4 mL) and anisole (100 µL) was cooled to 0 ° C and treated with trifluoroacetic acid (1.2 mL). The reaction was stirred at 0 ° C for 6.5 hours and the solvent was removed under reduced pressure. The oily residue is dissolved in 1 ml of acetone and added to 50 ml of n-hexane with stirring. The precipitate aa was filtered off and dissolved in 0.5 ml of ethyl acetate and to this solution was added 0.9 mg (0.054 mmol) of 2-ethyl sodium n-hexanoate. The precipitate was collected and dried to give 7 mg (19%) of a white solid.

NMR spectrum (deuterated water):

2.30 (α) 6H (2 OAc), 3.30, 3.60 (dd, 3-18 Hz) 2H (CH ₂ S), 3.82 (s) 3 H (OCH ₃ ), 4.80 ( d, 3, 12 Hz) 1H (1/2 _CH2 0), 5.06 (m) 2H (1/2 CHgO, C6), 5.66 (d, 3-6 Hz) 1H (C7), 6 84 (α) 1H (thiazole), 7.91 (s) 1H (Ar).

Example 34

Preparation of diphenylmethyleether / (6R-Trana) -3 - [//] 3,4-bia (acetyloxy) phenyl (amino) carbonyl / oxy / methyl) -7 - [(1,1-dimethylethoxy) carbonyl] amino] -8-oxo-5-thla-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid

To a solution of 3,4-bis- (acetyloxy) phenylphenyl cyanate (160 mg, 0.8 mmol) in absolute methylene chloride (7 mL) was added pyridine (0.2 mL) followed by 198 mg (0.4 mmol) of diphenylmethyl ester (6R-trans) -3 - (hydroxymethyl) -7 - [(1,1-dimethylethoxy) carbonyl] amino-8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid at a temperature room and under argon. The reaction mixture was stirred for 5 hours and then the solvent was removed under reduced pressure. The residue was taken up in ethyl acetate and the organic solution and washed with dilute hydrochloric acid and washed with anhydrous sodium sulfate. The crude product was chromatographed on silica gel using 1: 1 ethyl acetate / n-hexane as eluent to give 260 mg (84%) of a white solid.

NMR Spectrum (CDCl3) m.p.

1.48 (α) 9H (1H-bu), 2.28 (e) 6H (2-OAc), 3.43, 3.64 (dd, 3 · 18 Hz) 2H (CH ₂ S), 4, 84, 5.18 (dd, 3 · 12 Hz) 2H (CH 3 O), 4.96 (d, 3 · 6 Hz) 1H (C 6), 5.68 (dd, 3 · 6 Hz, 3 10 Hz) 1H (C7), 5.23 (d, 3-10 Hz) 1H (NH), 6.62 (s), 1H (thiazole, Ar), 7.0 (α) 1H (CH Ar ₂ ), 7.14 (α) 2H (Ar), 7.36 (m) 1H (Ar).

IC spectrum (KBr technique): 3340, 1773, 1722, 1530, 700.

Example 35

Preparation of the monosodium salt of (6R-) 6 (X), 7 (J) (3) - 3 - ///// 3,4-bis (acetyloxy) phenyl / amino / carbonyl / oxy / methyl) -7- // (2-Amino-4-thiazolyl) (methoxyimino) acetyl / amino] -8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid

A solution of 233 mg (0.31 mmol) difenylmathyleeteru / (6R-trans) -3 - ///// 3,4-bis (acetyloxy) _phenoxy-yl / amino / carbonyl / oxy / methy1 / -7 - // (1,1-dimethylethoxy) carbonyl / amino) -8-oxo-5-thia-1-pabicyclo [4.2.0] oct-2-ene-2-carbonyl acid from Example 34 in 5 ml of absolute 1, The 2-dichloroethane was cooled to 0 ° C in an ice / water bath and then treated with 0.6 mL of anionic acid and 4 mL of trifluoroacetic acid. The reaction mixture was stirred at 0 ° C for 5 hours and the volatiles were removed by evaporation under reduced pressure. Anhydrous diethyl ether was added to the residue and the precipitate was filtered off. The solid (130 mg) in 10 ml of absolute methylene chloride and 0.1 ml of triethylamine and at 0 ° C added to 106 mg (0.33 mmol) of S- (2-benzothiazolyl) - (Z) 1-aminothio-4-thiazolglyoxylate and 5 ml acetone. The mixture was stirred under argon at room temperature for 16 hours. The solvents are removed under reduced pressure and the residue is added to 5 ml of othylocotate and 10 ml of a 1% aqueous solution of sodium bicarbonate. 39 mg (20% of theory) of the desired product ·

NMR spectrum (perdeuterated dimethyl sulfoxide):

2.23 (s) 3 H (OAc), 2.26 (s) 3 H (OAc), 3.24, 3.50 (dd, 0-18 Hz) 2H (CH 3 S), 3.84 (s) 3 H ( OCH ₃ ), 4.90, 4.98 (dd, 0 · 12 Hz) 2H (OCH ₂ ), 5.0 (d, 3 · 6 Hz) 1H (C6),

5.60 (dd, 38 Hz, 31 Hz) 1H (C7), 6.78 (s) 1H (thiazole), 7.18 (d, 38 Hz) 1H (Sr), 7.32 (dd) 3 · 3 Hz, 38 Hz) 1H (Ar), 7.44 (d, 3 - 3 Hz 1H (Ar), 7.30 (broad d, 3 - 10 Hz), 1H (NH) ·

IR (KBr): 3330, 1762, 1672, 1732 (shoulder), 1612.

Example 36

Preparation of mono-sodium aoli [6R- / 6o (1,7 / J (Z)] - 3 - ///// 3,4-bis (acetyloxy) phenyl / amino / carbonyl / oxy / methyl] -7 - /// 2-amino - [(2-amino-2-oxoethoxy) imino] -4-thiazolyl] acetyl] amino-8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene -2-carboxylic acids

Following the procedure and conditions described in Example 38, 233 mg (0.31 mmol) of diphenylmethyl ether ((6R-trans) -3 - [(3R) -4-b8 (acetyloxy) phenynyl] amino] was reacted. carbonyl / oxy (methyl) -7 - [(1,1-dimethylsthoxy) carbonyl] amino-8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid of the acid (from Example 34) in 5 ml of absolute 1,2-dichloroethane and 0.6 ml of anisole are treated with 4 ml of trifluoroacetic acid, and then 87 mg (0.22 mmol) of O- (carbamoylmethyl) oxide S- ( 2-benzothiazolyl) -2-aminothio-4-thiazolglyoxylate. Purification by reverse phase chromatography (0-20% acetonitrile in water) gave 48 mg (23% of theory) of a white solid.

NMR spectrum (perdeuterated dimethyl sulfoxide):

2.23 (s) 3H (OAc), 2.26 (s) 3H (OAc), 3.24, 3.52 (dd, 3 · 16 Hz) 2H _(CH2 S), 4.42 (s) 2H _(CH2 CON), 4.93 (s) 2H (0CH _2), 5.03 (d, 3-6 Hz) 1H (C6), 5.64 (dd, 3 «6 Hz, 10 Hz · □ 1 H (C 7), 6.87 (s) 1 H (thiazole), 7.17 (d, 0 9 Hz) 1 H (Ar), 7.32 (dd, 3 Hz, 3 - 9 Hz) 1 H (Ar) ), 7.35 (ěiroký в) 2 H (NH _2), 7.42 (d, 3 · 3 Hz) 1H (Ar), 9.75 (d, 3-10 Hz) 1H (NH), 10.15 (broad β) 1 H (NH).

Example 37

Preparation of dimethyl ethyl ester ((6R-trane) -3 - [(3,4-b6 (acetyloxy) benzoyl) thio) methyl] -8-oxo-7 - [(phenoxyacetyl) amino-5-thia-1-azabicyclo] 4,2, O / oct-2-ene-2-carboxylic acid

According to the procedure and conditions described in Example 4, 230 mg (0.832 mmol) of 3,4-bis (acetyloxy) benzenecarbothioic acid sodium salt in 5 ml of di-methylformamide was reacted with 411 mg (0.775 mmol) of 1,1-dimethylethyl ether [6R- (6 *, 7) (Z) -3-iodomethyl) -7 - [(phenoxyacetyl) amino] -8-oxo-5-thia-1-azabicyclo [4.2.0] oct- Of 2-en-2-carboxylic acid in 5 ml of dimethylformamide, the reaction mixture is stirred for 2.5 hours and chromatographed on silica gel (methylene chloride / ethyl acetate 9: 1) to give 206 mg (40%) of a white solid. ·

NMR (CDCl3):

1.44 (s) 9H (t-Bu), 2.42 (s) 6H (2 OAc), 3.35, 3.61 (dd, 0 to 18 Hz) 2H _(CH2 S), 4.00 ,

4.29 (dd, 3 to 12 Hz) 2H (CHgSCO), 4.55 (s) 3H _(OCH3), 5.01 (d, 3 «6 Hz) 1H (C6),

5.91 (dd, 36 Hz, 3-10 Hz) 1H (C7), 6.90 (d, 3-8 Hz) 1H (Ar), 7.00 (m) 1H (Ar),

7.2, 7.3 (m) 3 H (Ar), 7.79 (s) 1 H (Ar), 7.86 (d, 3-8 Hz) 1 H (Ar).

IR (KBr) 3320, 1780, 1712, 1692, 1662 ·

UV spectrum (ethanol): λ _max 248 nm (£ 17,000),

.Lambda.max ^{nm 2X4 8S} ° ¹⁾ '

CS 271 490 B2

Example 38

Preparation of (6R-trans) -7-amino-3 - [(3,4-bls) (acyloxy) benzoyl (thio) methyl] -8-oxo-5-thia-1-azabicyclo [1,1-dimethyl-ethyl] monohydrochloride (4) 2, O / oct-2-en-2-ene-2-carboxylic acid

Following the procedure and conditions described in ee in Example 16 ee 147 mg (0.223 mmol)

(6R-trans) -3 - [(3,4-bis (acetyloxy) benzoyl) thiomethyl] -8-oxo-7 - [(phonoxyacyl) amino] -5-thia-1, 1-dimethylathylaether (6R-trans) -azabicyclo [4.2.0] oct-2-an-2-carboxylic acid (from Example 37) in 0.7 mL of methylene chloride was added to 27 µL (0.333 mmol) of pyridine and 61.7 mg (0.296 mmol) of chloride 70.5 mg (56% of theory) are obtained in the form of a white powder: NMR spectrum (perdeuterated dimethylsulfoxide):

1.53 (s) 9H (1H + -bu), 2.30 (s) 3H (OAc), 2.31 (s) 3H (OAc), 3.54, 3.83 (dd, 3-18 Hz) 1 H (CH ₂ S), 3.96, 4.33 (dd, □ - 12 Hz) 2H (CH 2 O), 5.17 (d, 0 6 Hz) 1 H (C 6). 5.22 (d, J = 6 Hz) 1H (C7), 7.48 (d, J = 8 Hz) 1H (Ar), 7.84 (s) 1H (Ar), 7.88 (d, J) 8 Hz) 1H (Ar) ·

IR (KBr): 1780, 1712, 1668.

UV spectrum (ethanol): λ _max 277 nm (£ £ 16,760).

Example 39

Preparation of monosodium aoli / 6R- / 60C, 7 /? (Z) - 3 - [(3,4-dihydroxybenzoyl) oxy] methyl-7 - [(2-amino-4-thiazolyl)] (2-amino-2-oxoethoxy) imino (acetyl) amino] -8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid.

It was reacted according to the procedure and conditions described in Example 20

350 mg (0.477 mmol) of 1,1-dimethylethyl ester [6R- [6,7,7] (Z)] - (3 -) - 3,4-bis (acetyloxy) benzoyl (oxy) methyl] -7- (2-Amino) - ((2-amino-2-oxoethoxy) imino) -4-thiazolyl (acetyl) amino) -8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2 -en-2-carboxylic acid in 5 ml methylene chloride and 500 µl anieol β 5 ml trifluoroacetic acid. The reaction mixture was worked up in the usual manner, to the residue were added ethyl acetate, methanol and saturated sodium bicarbonate solution (1: 1). After purification by chromatography and reverse phase (0-20% acetonitrile in water), 165 ml (54% of theory) were obtained. white solids.

NMR Spectrum (Psd-dimethylene sulfoxide):

3.56 (d, □ 18 Hz) 1H (1/2 CH ₂ S), 4.41 (s) 2H (CH ₂ ), 4.89, 5.17 (dd, □ 12 Hz)

2H (CH ₂ OCO), 5.05 (d, 0 → 6 Hz) 1H (C 6), 5.64 (dd, □ · 6 Hz, □ · 10 Hz) 1H (C7),

6.77 (d, □ 8 Hz) 1H (Ar), 6.84 (s) 1H (thiazole), 7.12 (э) 1H (OH, 7.299 (d, □ 8 Hz) 1H (Ar), 7 34 (s) 1 H (Ar), 7.50 (e) 1 H (OH), 9.75 (d, □ 8 Hz) 1 H (NH).

IR (KBr): 1762, 1672, 1608

UV spectrum (water): _{λ m} 218 nm (€ 900),

261 261 nm (C · 14 360).

г max '

HRMS (Ν + H) for ^C 22 ^H 21 ^N 6 ° 10 ^S 2 ^Na calculated 616.0614 found 616.0668.

Example 40

1,1-dimethylethyl ether / 6R- / 6 (X, 7 / J (Z)) - 3 - /// 3,4-bie (acetyloxy) benzoyl (thio) methyl) -7 - /// (2- amino-2-oxo-ethoxy) imino [(2-amino-4-thiazolyl) acetyl] amino-8-oxo-5-thiazabicyclo [4.2.0] oct-2-sn-2-carboxylic acid

Using procedure 2a of the conditions described in Example 18, from 243 mg (0.434 mmol) of (6R-trans) -3- [3,4-bis (acetyloxy) benzoyl] thio / methyl ethyl 1,1-dimethylethyl ester monohydrochloride (7-Amino-8-oxo-6-thla-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid and 158 mg (0.400 mmol) of O- (carbamoylmethyl) oxime S- ( 2-benzothiazolyl) -2-aminothio-4-thiazolylglyo

CS 271 490 B2 xylate in 8 ml of dimethylformamide and 16 ml of methylene chloride gave 180 mg of a white solid (after chromatographic purification using methylene chloride containing 0 to 50% methanol as eluent).

NMR Spectrum (CDCl3):

1.57 (s) 9H (b-Bu), 2.31 (s) 6H (2 OAc), 3.39, 3.63 (dd, □ 18 Hz) 2H _(CH2 S), 3.98, 4.36 (dd, Э 15 Hz) 2H _(CH2 SC0), 5.07 (d, □ · 6 Hz) 1H (C6), 5.60 (br) 2H _(NH2), 5.95 (dd Hz6 Hz, □ 10 Hz) 1H (C7), 6.85 (s) 1H (thiazole), 7.30 (d, □ · 8 Hz) 1H (Ar), 7.80 (d, □ 2 Hz) 1 H (Ar), 7.88 (dd, □ 2 Hz, □ 8 Hz) 1H (Ar), 8.32 (broad) 1H (NH).

IR (KBr): 1778, 1710, 1672.

UV spectrum (ethanol): λ _max 240 nm (# 20,750) λ _max 278 nm (<C 17,300)

Mass spectrum: m / z 749 (M + H).

Example И1

Preparation of monosodium eoli [6R- / 6 (X, 7R (Z)] - 7 - [] 2-amino] (2-amino-2-oxoethoxy) imino-4-thiololyl / Aoatyl / eMno / 3- // (1-Dihydroxybenzenesoyl) thi (methyl) -O-oxo-5-thia-1-aabicyclo [4.2.0] oct-2-ene-2-carboxylic acid

Using the procedure and conditions described in Example 20, ee was reacted with 305 mg (0.408 mmol) of 1,1-dimethylethyl ether (6R- / 6X, 7/7 (2) // - 3 - /// 3,4-). bie (acetyloxy) benzoyl (thio) methyl-7 - [(2-amino-2-oxyethoxy) imino] (2-amino-4-thiazolyl) acetyl] amino] -8-oxo-5-thia-1- azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid (from Example 40) in 6 ml of methylene chloride and 600 µl of anisole with 13 ml of trifluoroacetic acid and the residue was stirred with a mixture of ethyl acetate, methanol and saturated bicarbonate solution. NaCl in a 1: 1: 1 ratio. After chromatographic purification using reverse phase chromatography (0-20% acetonitrile in water), ee yielded 200 mg of a white solid.

NMR spectrum (perdeuterated dimethylsulfoxide):

3.09, 3.51 (dd, 0-18 Hz) 2H (CH ₂ S), 4.03, 4.18 (dd, □ »12 Hz) 2H (CHgSCO), 4.41 (з) 2H ( OCH ₂ ), 5.00 (d, □ - 6 Hz) 1H (C 6), 5.59 (dd, □ - 6 Hz, □ - 10 Hz) 1H (C7),

6.79 (d, □ 8 Hz) 1H (Ar), 6.83 (s) 1H (thiazole), 7.09 (α) 1H (OH), 7.30 (m) 6H (Ar, 2NH ₂ ) 7.48 (s) 1 H (OH), 9.73 (d, 0-10 Hz) 1 H (NH).

IR (KBr): 1760, 1672, 1592, 1162, 1088.

UV spectrum (water): λ _max 230 nm (£ 16,850),

Amax ²⁸² 9 ¹² ∞) HRMS (m, 4 H) for C ₂₂ ^H 19 ^N 6 ^S 3 ° 9 ^Na: calculated 631.0375 found 631.0362.

Example 42

Preparation of 1,1-dimathylethyleatar [6R- (6 ', 7') (2) H] -3- [3,4-bile (acetyloxy) benzoyl] oxy (methyl) -7- (2-amino-) 4-thiazolyl) [2- (1,1-diphenyl-ethoxy) -2-oxo-ethoxy] -amino (acetyl) -amino] -8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2 -en-2-carboxylic acids

Following the procedure and conditions described in Example 18, (6R-trans) -7-amino-3 - [(3,4-bie) -benzyl] -ethyl-ester of 1,1-dimethylethyl ester, 1,20 g (2,37 mmol) was used. (acetyloxy) benzoyl (oxy) methyl] -8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid 46 mL methylene chloride and 1.0 g (222 mmol) 2 - [1- (2-amino-4CS) 271 490 B2 1,1-dimethylethyl ester

-thiazolyl) -2- (benzothiazol-2-ylthio) -2-oxoethyl / imino / oxy / acetic acid in 46 ml of methylformamide and 23 ml of methylene chloride after stirring for four hours and after chromatography Purification on silica gel using methylene chloride / ethyl acetate 8: 2 gives 920 mg of white solid ·

NMR Spectrum (CDCl3):

1.42 (s) 9H (D-α), 1.55 (β) 9H (t-bu), 2.31 (s) 6H (2 OAc), 3.41, 3.62 (dd, 3 18) Hz) 2H _(CH2 S), 4.73, 4.77 (dd, 3 to 17 Hz) 2H _(CH2 0C0Ar), 5.06, 5.39 (dd, 3 · 16 Hz) 2H (0CH ₂ C0), 5.09 (d, 3-7 Hz) 1H (06), 5.97 (dd, 37 Hz, 3 · 10 Hz) 1H (C7), 7.03 (s) 1H (thiazole), 7.29 (d, 3-8 Hz) 1H (Ar), 7.85 (d, 3-2 Hz) 1H (Ar), 7.95 (dd, 3.2 Hz, 3-8 Hz) 1H. (Ar), 8.63 (d, 3-10 Hz) 1 H (NH).

IR (KBr) 3400, 1780, 1722, 1682.

UV spectrum (ethanol): _λflax 236 nm (£ - 27,500)

Mass Spectrum: m / z 790 (M4H) ·

Example 43

Preparation of the disodium salt of [6R- (60G) 7Z- (Z)] - 7 - [(2-amino-4-thiazolyl)] (carboxymethoxy) imino / acetyl / amino] -3- [3,4-dihydroxybenzoyl) oxy (methyl) -8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid

Using the procedure and conditions described in Example 20, by reaction of 920 mg (1.16 mmol) of 1,1-dimethylethyl ester (6R- / 6 ', 7' (Z)) - 3 - /// 3, 4-bie (acetyloxy) benzoyl (oxy) methyl] -7 - [(2-amino-4-thiazolyl)] (2- (1,1-dimethylethoxy-2-oxoethoxy) amino) 1-Amino-8-oxo-6-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid in 24 ml of methylene chloride and 2.3 ml of anisole 8 with 24 ml of trifluoroacetic acid and subsequent stirring of the residue with a mixture of ethyl acetate, methanol and saturated sodium bicarbonate solution and chromatographic purification using reverse phase (0 to 30% acetonitrile in water) gave 385 mg (62%) of a white solid.

NMR spectrum (deuterated water):

3.56 (dd, 3 to 18 Hz) 2H _(CH2 S), 4.42 (s) 2H (CHgCOgNa), 4.67, 4.96 (dd, 3-15 Hz), 2H (CH ₂ 0) 5.07 (d, 3-6 Hz) 1H (C6), 6.68 (d, 3-6 Hz) 1H (C7), 6.71 (d, 38 Hz) 1H (Ar) 6.87 (s) 1H (thiazole), 7.30 (m) 2H (Ar).

IR (KBr): 1755, 1650, 1602 ·

UV spectrum (water): _λmax 203 nm (δ - 17,600),

231тах ²³¹ (- - 15 400), Amax ^{260 nm 9} θ ⁰⁰ ) · Amax ^{302 nm 8 000)} '

Example 44

Preparation of (6R-trans) -7- [(2-amino-4-thiazolyl)] - 1- (aminocarbonyl) -1-methylethoxy (imino) acetyl / amino] -3-dimethylatyl ester 1,1-dimethylatyl ester 4-bis (acetyloxy) benzoyl (oxy) methyl] -8-oxo-6-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid

A solution of 542 mg (1.01 mmol) of (6R-trans) -7-amino-3 - [(3,4-bis (acetyloxy) benzoyl) oxy] methyl] -8-oxo-5 1,1-dimethylethyl ester monohydrochloride -tla-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid in 25 ml of methylene chloride was washed twice with 25 ml of a 1: 4 mixture of saturated sodium bicarbonate and brine. The organic solution was dried over sulfate. magnesium and filtered with DC. To the filtrate was added a solution of 411 mg (1.00 mmol) of 2,2-amino-DC - [(2-amino-1,1-dimethyl-2-oxoethoxy) imino] -4-thiazoleacetic acid, 136 mg (1, 00 mmol) of N-hydroxybenzotriazole and 211 mg (1.00 mmol) of dicyclohexylcarbodiimide in 25 ml of absolute dimethylformamide.

CS 271 490 B2 chromatography on silica gel gave 320 mg (42% of theory) of the title compound.

NMR Spectrum (CDCl3):

1.50 (s) 6H (2 _CH3), 1.54 (в) 9H (t-Bu), 3.42, 3.61 (dd, J = 18 Hz), 5.06 (m)

2H (1/2 CH ₂ O, C 6), 5.32 (d, J = 13 Hz) 11 H (1/2 CH ₃ O), 5.90 (dd, J = 5 Hz, J = 9 Hz)

1H (C7), 6.40 (broad s) 2H (NH3), 6.75 (s) 111 (thiazole), 7.28 (d, < J = 8 Hz) 1П (Ar), 7.83 (d) J = 2 Hz) III (Ar), 7.93 (dd, J = 2 Hz, J = 8 Hz).

IR (KBr): 1780, 1722, 1680, 1532.

UV spectrum (ethanol): λ _max 236 (δ = 29050).

Mass spectrum: m / z 761 (M + H).

Example 45

Preparation of (6R-trans) -7 - [(2-amino-4-thiazolyl)] - 1- (aminocarbonyl) -1-methylethoxy / imino / acetyl / amino] -3 - [(3,4- dihydroxybenzoyl) oxy (methyl) -8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid.

320 mg (0.421 mmol) of (6R-trans) -7 - [(2-amino-4-thiazolyl)] - 1, 1-dimethylethyl ester (6R-trans) -1 was reacted according to the procedure and conditions described in Example 20 - (aminocarbonyl) -1-methylethoxy / imino / acetyl / amino / -3 - [(3,4-bis (acetyloxy) benzoxyl) oxy] methyl] -8-oxo-5-thia-1-azabicyclo [4] > 2.0 / oct-2-ene-2-carboxylic acid in 5 ml methylene chloride and 500 µl anisole with 5 ml trifluoroacetic acid. The reaction mixture was stirred for 3 hours and a mixture of ethyl acetate, methanol and saturated sodium bicarbonate solution was added to the residue. Purification by reverse phase chromatography (0 to 30% acetonitrile in water) gave 123 mg (45%) of a white solid.

NMR spectrum (deuterated water):

1.60 (s) 6H (2 _CH3), 3.53, 3.78 (dd, J = 18 Hz) 2H (CHGS), 4.93, 5.17 (dd, J =

Hz) 2И (CH ₂ O), 5.28 (d, J = 4Hz) 1H (C6), 5.90 (d, J = 4Hz) 1H (C7), 6.88 (d, J = 8Hz) Hz) 1 H (Ar), 7.12 (s) 1 H (thiazole), 7.50 (s) 1 H (Ar), 7.55 (d, J = 8 Hz) 1 H (Ar).

IR (KBr): 1762, 1672, 1605.

UV spectrum (water): λ _max 218 nm (δ ^s 22,300).

HRMS (M + H) for ^C 24 ^H 23 ^N 6 ° 12 ^S 2 ^Na: calculated 643.0947 found 643.0893 ·

The following example illustrates a pharmaceutical formulation which contains as active ingredient a cephalosporin derivative obtained by the process of the invention.

Example A.

Preparation of dry ampoules for intramuscular administration

1 g of the monosodium salt of [6R- (6 [(7) (Ζ)] - 3 - [(3,4-dihydroxybenzoyl) oxy] methyl] -7- [2-amino-4] is prepared in a conventional manner. -thiazolyl) [(2-amino-2-oxoethoxy) amino] acetyl amino] -8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid and charged The sterile water ampoule contains 10% propylene glycol and 2.5 ml of a 2% aqueous lidocaine hydrochloride solution is added to the lyophilisate.

CS 271 490 B2

SUBJECT OF THE INVENTION

Claims (14)

A process for the preparation of novel acyl derivatives of the general formula I (I) in which X is -COCO-, -SCO-, -NHCO- or -OCONH-, is an acyl group that is used to acylate p-lactam antibiotics and that contains up to 21 carbon atoms, for example, the group of formulas ^H 101 ^-CH 2 ^C ° N-CONH-CH-CO- R _13O -ON = C wherein the CTI-101 ^represents a 5-membered heterocyclic ring containing 1 or 2 nitrogen atoms and / or sulfur atoms and which is optionally substituted by amino, ^R 120 ^is an alkyl group having 1 to 8 carbon atoms and R ₁₃₀ is C 1 -C 8 alkyl optionally substituted by carboxy, including carboxy in salt form, carbamoyl or C 2 -C 9 alkoxycarbonyl, hydrogen or C 1 -C 8 alkoxy formulas CS 27 1 4 90 B2 or CS 271 490 B2, in which ^R 4 ^'R 4' and says ^znaraena independently hydrogen or -ΟΟβ θθ ^ where ^ 200 ^ ^represents straight-chain or branched alkyl having 1 to 8 carbon atoms, А а В are halogen atoms, a, b, x, у and z are independently zero and in this case a hydrogen atom is present at the appropriate position of the ring, or they are number 1 with at least two of y, x and z being the number 1 and m represents zero or an integer from 1 to 8, as well as the pharmaceutically usable salts of these compounds and the hydrates of the compounds of formula I or their salts, characterized in that from the ester of formula XIX (XIX) X, R ^, R? and R-j are as defined above and R represents a carboxyl protecting group which is used in the chemistry of cephalosporins and which can be readily cleaved under mild conditions, for example by a tert-butyl group, a p-nitrobenzyl group or an allyl group, cleaves a protecting group R, preferably a p-nitrobenzyl group R by hydrolysis in in the presence of sodium sulfide, a tert-butyl group by reaction with trifluoroacetic acid in the presence of either a sol or an allyl group by palladium catalyzed transallylation in the presence of a sodium or potassium salt of 2-ethylhexanoic acid, then optionally recovering the product as salt, hydrate or hydrate. 2. A process according to claim 1, wherein the corresponding compounds of formula (XIX) are used as starting materials to give compounds of formula (I) wherein m is 0 or 1 and the other substituents are as defined in 1. 3. Process according to claim 1 or 2, characterized in that the corresponding compounds of the general formula (XIX) are used as starting materials to give compounds of the general formula (I) in which R @ 1, R @ 2 and R @ 4 are independently hydrogen. or -COCH3 and the other substituents are as defined in 1 or 2 above. 4. A process according to any one of claims 1 to 3, wherein the corresponding compounds of formula (XIX) are used as starting materials to produce compounds of formula (I) wherein A and V are chlorine and the other substituents are as defined in one of 1 to 3. Process according to one of Claims 1 to 4, characterized in that the corresponding compounds of the general formula (XIX) are used as starting materials to give the compounds of the general formula: CS 271 490 B2 of formula I in which R g represents a hydrogen atom and the other substituents have the meanings given in one of points 1 to 4; Process according to one of Claims 1 to 5, characterized in that the corresponding compounds of the general formula (XIX) are used as starting materials to give compounds of the general formula (I) in which X represents a group -ОСО- and the other substituents have the meaning given in from points 1 to 5 · 7. A process as claimed in any one of claims 1 to 5, wherein the corresponding compounds of formula (XIX) are used as starting materials to give compounds of formula (I) wherein X is -SCO- and the other substituents are as defined in points 1 to 5 « Method according to one of Claims 1 to 5, characterized in. Use of the corresponding olefins of formula XIX as the starting material to form compounds of formula I wherein X is -NHCO- and the other substituents are as defined in one of items 1 to 5. 9. A method as claimed in any one of Claims 1 to 9, wherein the compound of formula (XIX) is used as a substance to produce compounds of formula (I) wherein X is -OCONH- and the other substituents are as defined in one of points 1 to 5. 10. A process according to any one of Claims 1 to 9, wherein the corresponding compounds of formula (XIX) are used as starting materials to produce compounds of formula (I) wherein the acyl group is represented by the formula and the other substituents are as defined in points 1 to 9. Process according to one of Claims 1 to 9, characterized in that the corresponding compounds of the general formula (XIX) are used as starting materials to give the compounds of the general formula (I) in which the heteroaromatic group of the general formula R ₁₀₁ -CH ₂ -CO- R ₁₀₁ represents a 5-membered heterocyclic ring containing 1 or 2 nitrogen and / or sulfur atoms, which is optionally substituted with amino, and the other substituents are as defined in one of items 1 to 9. Process according to one of Claims 1 to 9, characterized in that the corresponding compounds of the general formula (XIX) are used as starting materials to give the compounds of the general formula (I) in which R 1 represents //( 4 - sub-2 - 2,3 -). a dioxo-1-piperazinyl) carbonyl (amino) arylacetyl group of the formula CS 271 490 B2 where ^ 120 ^znaraeaa alkyl group having 1 to 8 carbon atoms and the other substituents are as defined in one of claims 1 to 9 · Process according to one of Claims 1 to 9, characterized in that the corresponding compounds of the general formula (XIX) are used as starting materials to give compounds of the general formula (I) in which R 1 represents a group of the general formula ^R 130 -O- N- C- CO-, ^E 101 wherein ^R 101 ^is = 5-membered heterocyclic ring containing 1 or 2 nitrogen atoms and ^a sulfur or, which is optionally substituted amino group and an alkyl group having 1 to 8 carbon atoms which is optionally substituted with carboxy, carboxy including in the form of a salt, a carbamoyl group or a (C 2 -C 9) alkoxycarbonyl group, and the other substituents are as defined in one of items 1 to 9. 14. The process of claim 13, wherein the corresponding compounds of formula (XIX) are used as starting materials to form the compounds of formula (I) as defined in (13), wherein the acyl group R &lt; 1 &gt; to 8 carbon atoms or a group of the formula COP where