DE2911787C2 - Carbacephem derivatives and their uses - Google Patents

Description

O
-CH-OCR ₄

R ₃

means,

where R represents a hydrogen atom or an alkyl radical having 1 to 5 carbon atoms and R _{4 represents} an alkyl radical having 1 to 6 carbon atoms, an alkoxy radical having 1 to 6 carbon atoms or a phenyl group, and their salts.

2. (±) -cis-7y8- [2- (2-Amino-4-thiazolyl) -2-syn-methoxyiminoacetamido] -4ff-methyl -! - azabicyclo [4.2.0] oct-2-en- 8-one-2-carboxylic acid.

3. (±) -cis-7- [2- (2-Amino-4-thiazolyl) -2-syn-methoxyiminoacctamido] -l-azabicyclo [4.2.0] oct-2-en-8-one- 2-carboxylic acid.

4. Use of the compounds according to Claims 1 to 3 for the treatment of bacterial infections.

The nomenclature of the 7-acylamino-1-carba-3-cephem-4-carboxylic acid derivatives according to the invention of the general Formula I is in J. AM. Chem. Soc, 96: 7584 (1974).

Carbacephem VerbitWüngcn with a substituted methyl group in the 3-position, such as (±) -l-carbacephalotin of the formula ά

f 1

^. r ν

1 IIII IX f

COOH

are known. Antibacterial efficacy has also been reported in some of them (cf. the aforementioned I. Druckschrift and J. Med. Chem., Vol. 20 (1977), p. 551). |

In DE-OS 27 16 707 lactams are of the formula

H ₂ NR ₄

ι I

COORj I

described for the preparation of cephem derivatives of the formula

COOR ₃

be used.

J !; H ^ rLm inpnc, ^ ,, nf ¹ ' ^{m where X is a} methylene group and A is a hydrogen

f ° ^m ^ ₇ 'JV' f ^r ^ ^E -. ² J ^{16 707 In no} case as practical examples described ° ^S In the DE-OS 27 16707, neither the description is a method for preparing such compounds nor description her. A review of the literature shows that all known carbacephem compounds have an optionally substituted methyl group on the CI carbon atom (cf. J. Am.

Γ iTJSn? rr, ⁰ K r, ⁷³⁸⁴ ' ^J v ^e K- ^Chem -' ^Vol . ^{20 (1977)} < ^{S 551 and DE} - ° ^S 23 55 209). One of these compounds substituted on the £ Ι ιΛβη ~ u ^d ° ^{m is} also in the list of compounds in the DE-OS

The preparation of the carbacephem compounds of the above structure by Merck's total synthesis is described, for example, in "Progress in Medicinal Chemistry", Vol. 14 (1977) pp 198-212 Preparation of 3-H-cephem compounds is from HeIv. Chim. Acta, Vol. 57 (1974) pp 2044-2054

? ^{er E} _c ^rr ' ^nd » ⁿ 8 !! ⁶ S ^{1 di} _Ä ^e levy to create new Carbacephemderivaic with good antibacterial effectiveness. This object is achieved by the invention.

The invention relates to carbacephem derivatives of the general formula I

10

, 5

R ₃

(D

in A ₁₁ a hydrogen atom e an alkyl radical with 1 to 6 carbon atoms. Alkenyl radical with 2 to 6 carbon atoms, alkynyl radical with 2 to 6 carbon atoms or a cycloalkyl radical with 3 to 6 carbon atoms or a cycloalkyl radical with 3 to 6 carbon atoms, the radicals optionally being replaced by a halogen atom, a carboxyl, cyano or carbamoyl group or a Nicderalkoxvcarbonvlrest are substituted by 1 to 4 carbon atoms;

R _{2 is} a hydrogen atom, an alkyl radical having 1 to 5 carbon atoms or an acyloxy radical having 1 to 5 carbon atoms and R _{1 is} a hydrogen atom or a radical of the general formula

O -CH-OCR ₄

means,

where R represents a hydrogen atom or an alkyl radical having 1 to 5 carbon atoms and R _{1 represents} an alkoxy radical having 1 to 6 carbon atoms, an alkoxy radical having 1 to 6 carbon atoms or a phenyl group and their salts. "

In the event that R ₁ in general formula 1 denotes a hydrogen atom or a free carboxyl group is present in the 7-substituent, the compounds of general formula I can be in the form of salts with inorganic or organic bases. They can also be present as salts of the amino group in the 7-substituent with inorganic or organic acids.

The compounds of the general formula I also include all stereoisomers with respect to the 6-7- and 4-positions and mixtures thereof. The stereoisomers which are in the cis configuration with respect to the 6- and 7-positions have a higher antibacterial activity than the corresponding trans isomers. Therefore the cis isomers are the more valuable antibiotics. The remainder = N0 A ₁₁ in the acyl residue can be in the form of two geometric isomers (syn and anti)

—C — coil

N-OA ₁₁

(syn)

- C-CO-

A ₁₁ ON
(anti)

50

55

60

65

are present. Since the syn isomer is superior to the anti isomer in antibacterial activity, the syn isomer is the more valuable antibiotic.
It is known that the thiazolyl group of the formula

rearranged reversibly to the thiazolinyl group, as indicated below. Both groups are in the alllu commonly regarded as identical. In the context of the invention, for the sake of simplicity, only the thiazolyl group is in each case specified.

1 ^ HN == <1

N-! J ν-y

Thiazolyl

Thiazoline vl

The compounds of the general formula I are prepared by the corresponding compound of the general formula ΙΊ acylated according to the following reaction scheme I:

Reaction scheme I.

Π COOH H ₂ N

^Ν Ύ ♦

OA ₁

[III]

[Π]

[I]

In Reaction Scheme I, R 1, R 1 and A _{n have} the meanings given above.

The acylation (condensation) is common in the field of penicillins and cephalosporins Way done.

The compounds of the general formula I are obtained by condensing a compound of the general Formula ΙΓ, its salt or a compound functionally equivalent to the 7-amino group with a Carboxylic acid of the general formula III or one of its reactive derivatives prepared, where appropriate subsequently the protective groups are split off in the customary manner.

As protecting groups for z. B. amino or carboxyl groups in compound III are used accordingly protective groups customary in the field of cephalosporins or penicillins. Such protecting groups are also below specified.

Suitable salts of the compounds of the general formula II are inorganic and organic salts, for. B. hydrochlorides, sulfates, carbonates, phosphates, formeates, trifluoroacetates and maleates. Examples of salts of the carboxylic acid of the general formula II, in which R ₁ denotes a hydrogen atom, are sodium, potassium, calcium and ammonium salts and salts with organic amines. The salts are prepared in the usual way.

Examples of compounds which are functionally equivalent to the compounds of the general formula II are the corresponding 7-monosilyl or 7-disilylamino derivatives.
Examples of reactive derivatives of the carboxylic acid of the general formula III are
Acid halides,
Acid anhydrides,
mixed acid anhydrides,
reactive esters,
reactive thioesters and
Acid azides.

The condensation (acylation) using the aforementioned derivatives is explained below.

a) Use of an acid halide

A 7-amino compound and a acid halide are in an inert solvent, preferably in Presence of a prolonger, condensed. The acid halide is prepared in the usual way.

Inorganic or organic bases, preferably /, are used as prolonenuk / .eplor. B. sodium carbonate, Potassium carbonate, sodium hydroxide, potassium hydroxide, triethylamine, N-methylmorpholine and pyridine.

Any solvent that does not impair the reaction can be used as the solvent. Preferably becomes water, an ether such as tetrahydrofuran and dimethoxyethane, a Kstcr such as ethyl acetate, an amide such as

i 55 a) J
i b) C) d) e) η

Dimethylacetamide and hexamethylphosphoric acid triamide. or a sullbxide. like DimeihylsuHbMd. or a mixture of at least two of these solvents is used.
The reaction takes place at temperatures from -20 to -4O ⁶ C, before / ugsu eise 0 ⁰ C to Raumlempcialui-, duivhiu-lülm.

b) Finsat / a.s Saurcunhydrids ν

A 7-amino compound and an acid anhydride are condensed in an inert solvent. That .Acid anhydride is produced in the usual way.

Any solvent that does not impair the reaction can be used as the solvent. Preferably will the same solvent as used in the method given above under (a). The applied Temperatures are also in the range of the aforementioned method.

A typical example of this type of reaction is the use of a carbodiimide, such as dicyclohexylcarbodiiinids.

c) Use of a mixed acid anhydride

A 7-amino compound and a mixed acid anhydride are condensed in an inert solvent. The mixed acid anhydride is prepared in a conventional manner. For example we have a corresponding Carboxylic acid of the formula III and a chloroformate, such as ethyl chlorineate or isobutyl chloroformate, implemented in the presence of a base

Any solvent that does not impair the reaction can be used as the solvent. In general becomes a non-aqueous solvent or a mixture of water and a non-aqueous one Solvent, preferably the same solvent as in the aforementioned methods, used. Of the The temperature range is also in the range that is also used for the aforementioned methods.

d) Use of a reactive η ester

A 7-amino compound and a reactive ester are condensed in an inert solvent. Solvent and reaction temperature correspond to the information given under (a) to (O) above.

A phenyl ester such as p-nitrophenyl ester or trichlorophenyl ester, for example, is used as a reactive ester a methyl ester with an electronegative group, such as a cyanomethyl ester, or an N-oxydiazyl. <» imidesler, such as an N-hydroxysuccinimide ester, is used.

The reactive ester is prepared in the usual way by adding a corresponding carboxylic acid and reacting a corresponding hydroxyl compound in the presence of a dehydrating condensing agent such as dicyclohexylcarbodiimide.

35 e) Use of a reactive thioester

This method is carried out according to the method given under (d). A preferred thioester is for example p-nitrothiophenyl ester used, which according to the method for the preparation of a mixed Acid anhydride or using dicyclohexylcarbodiimide is produced. -!O

f) Use of an acid azide

A 7-amino compound and an acid azide are dissolved in an inert solvent similar to those above condensed under (a) to (e) mentioned methods.

The acid azide is prepared in such a way that a hydrazide of a corresponding carboxylic acid with Nitrous acid reacts at -20 to 0 ° C.

In the event that the reactive derivative of the carboxylic acid used according to the invention, for example has an amino or carboxyl group which is amenable to acylation, these groups are preferred protected from condensation with the amino compound with suitable protective groups. As protecting groups For example, those used are common in the field of penicillins and cephalosporins are.

For example, the tert-butyloxycarbonyMBoc), benzyloxycarbonyl- (Cb /), Trichlorethyloxycarbonyl, trityl, formyl, chloroacetyl or trialkylsilyl group, protons and the like Residues of s-diketones or s-ketoesters are preferred.

Corresponding carboxyl protective groups are, for example, the tert-bulyl, benzyl, p-methoxybenzyl, p-nitrobenzyl or benzhydryl group.

The aforementioned protective groups are split off in the field of penicillins and cephalosporins usual way.

Examples of the splitting off of the amino protective groups are given below. ω

The Boc group is made using an acid such as formic acid, acetic acid, trifluoroacetic acid or hydrochloric acid, cleaved.

The Cbz group is formed by catalytic reduction or using a mixture of hydrobromic acid and acetic acid split off.

The trityl group is produced by catalytic reduction or using an acid such as trilluoroacetic acid (ö cleaved.

The formyl group is split off by hydrolysis using an acid or a base.

The chloroacetyigruppe is split off using thiourea.

A trialkylsilyl group is split off by hydrolysis. A proton is split off by neutralization.

The jJ-diketone or the, / f-ketoester is split off by acid hydrolysis.

Examples of the splitting off of carboxyl protective groups are given below:

The tert-butyl group is split off using an acid such as trifluorocacetic acid.

The benzyl or p-nitrobenzyl group is obtained by catalytic reduction or e.g. B. using a Lewis acid, such as aluminum chloride, split off.

The Benzhydryl- or p-Methoxyben7vlgruppe is by catalytic reduction or using z. B. Trilluoroacetic acid, a mixture of hydrogen bromide and acetic acid or a mixture of hydrogen chloride and methanol, split off.

The acylation reaction according to the invention can optionally be carried out by silylating the starting compound of the general formula II using a silylating agent such as trimethylchlorosilane base, hexamethyldisilazane or Ν, Ο-Bistrimethylsilylaeetamid, are favored to convert the starting compounds into organic Solubilize solvents and activate the amino group.

The starting compounds of the general formula II can be prepared in accordance with DE-OS 29 11 786. An example is given in the reaction schemes V and II 'below.

Reaction scheme Γ

20 R

OHCCH ₂ Ch-CH =

/ 1: R = H 30 V- R = CH

H ₂ NP (OEt) ₂

CO ₂ R ₁

NP (OEt) ₂

CO ₂ R ₁

: R = H \ ': R = CH., Y

N ₃

NjCH ₂ COCI

Close

CO ₂ R ₁

/ 5: R = H \ \ 5 ': R = CH ,;

Saponification

CO ₂ H

Et · ■ =

Π: R = H \ 1,7 ': R = CH _r J

Reaction scheme II '

COOR,

10

OCOCH ₃ H ₂ N

reduction

VYY

OCOCH ₃

COOR ₁

Saponification

Saponification

H ₂ N

OCOCH ₃

reduction

COOH

12th

OCOCH ₃

C0, H

In the reaction schemes Γ and ΙΓ the compounds 6,7,6 ', 7', 11 and 12 are examples of starting compounds of general formula II. The preparation of the starting compounds is in the following Reference examples described.

The compounds of general formula Γ (compound of general formula I, in whichR, a hydrogen atom means), can by splitting off the carboxyl protective group according to the reaction scheme below II are produced:

Reaction scheme II

In Reaction Scheme II, R 1 represents a carboxyl protecting group; Ri and A ₁ have the above meanings.

The saponification reaction takes place in the manner customary in the field of penicillins and caphalosporins.

As a reaction in which the radical -COOR is converted into a carboxyl group, for example the types of reactions

a) catalytic reduction,

b) acidolysis,

c) cleavage using a Lewis acid,

d) hydrolysis,

e) non-catalytic reduction and
Γ) Use of an esterase

in question. These methods are explained below.
5

a) Catalytic reduction

The remainder -COOR) is in the presence of a catalyst in a hydrogen atmosphere in an inert Solvent converted into a carboxyl group. Any solvent can be used as the solvent

the one that does not affect the response. Ethanol, water, tetrahydrofuran, dioxane, ethyl acetate or acetic acid or a mixture of at least two of these solvents is preferably used. As a catalyst z. B. palladium-on-carbon, platinum oxide, palladium-on-calcium carbonate and Raney nickel. The reaction is generally carried out at a pressure of 1 to 50 bar and a temperature of 0 to 100 ^° C., preferably at ambient pressure and room temperature.

This method is preferably used when R, for example a benzyl, p-nitrobenzyl, diphenylmethyl or p-Meihoxybenzylgruppe means.

b) acidolysis

The remainder -COORi is converted into a carboxyl group with an acid in an inert solvent. The acid is such. B. hydrogen chloride, p-toluenesulfonic acid or trifluoroacetic acid. As a solvent any solvent that does not interfere with the reaction can be used. Ethyl acetate is preferred, Benzene, ethanol, acetic acid, dioxane, methylene chloride or chloroform or a mixture of at least two of these solvents are used.

2s The reaction is carried out at a temperature of -15 to 50 ^° C., preferably 0 to 25 ^° C., in 10 minutes to 5 hours, preferably 30 minutes, to 3 hours.

c) cleavage using a Lewis acid

The radical -COOR ₁ is converted into a carboxyl group by cleavage reaction in the presence of a Lewis acid in an inert solvent. Any solvent that does not impair ^the reaction can be used as the solvent. Preferably, a mixture of a nitroalkane, such as nitromethane,

Ί and a haloalkane, such as methylene chloride, are used. As a Lewis acid, for. B. aluminum chloride, boron

trifluoride, titanium tetrachloride or tin tetrachloride can be used. The acid is added in amounts of 1.0 to 1.5 molar equivalents, based on the compound of the general formula I ". The reaction is preferably carried out in the presence of an agent such as anisole which accepts carbonium cations ⁰ C, preferably at room temperature, carried out for 1 to 10 hours.
sf d) hydrolysis

The radical -COOR is converted into a carboxyl group by hydrolysis in the presence of an acid or a base in an inert solvent. The acid can be, for. B. p-ToIuolsulfonsäure, hydrochloric acid or acetic acid _e can be used. Any solvent that does not impair the reaction can be used as the solvent.

Preferably z. B. a 2 percent aqueous Melhanoüösung, N, N-Dimethyiförmamid or a mixture

from acetic acid, water and tetrahydrofuran used. The reaction is conducted at einerTemperaturvonO to 50 ⁰ C, preferably 15 to 25 ° C, for 10 minutes to 2 hours is performed.

The base is preferably calcium carbonate in amounts of 1 to 6 molar equivalents, based on the compounds I ". Any solvent which does not impair the reaction can be used as the solvent. A mixture of tetrahydrofuran and water, dioxane and water or acetone and water is preferably used in each case The reaction is generally carried out at a temperature of from 0 to 30 ^° C. for 30 minutes to 24 hours.

e) Non-catalytic reduction

The remainder -COOR is converted into a carboxyl group by reduction in an inert solvent. For example, zinc can be used together with an acid for the reduction. As a solvent z. B. acetone, water, dioxane, tetrahydrofuran, ethanol, acetonitrile, Ν, Ν-dimethylformamide or acetic acid or a mixture of at least two of the solvents mentioned. The acid can be, for. B. hydrochloric acid, formic acid or acetic acid can be used. The reaction is conducted at a temperature of 0 to 100 ⁰ C.

preferably 0 to 40 ⁰ C, are carried out for 1 to 10 hours. The amount of zinc used is generally 1 to 10 molar equivalents.

If suitable reactants are used, a protective group in the 7-substituent can also be split off at the same time as the ester saponification.
The compounds of general formula I in which R ₁ denotes a hydrogen atom have an excellent

Outstanding antibacterial effectiveness against Gram-positive and Gram-negative bacteria, which is why they can be used as anti-infectious or antibacterial agents. The compounds of the general formula I in which R _{1 is} not a hydrogen atom can also be used as starting compounds in order to prepare the compounds of the general formula 1 in which R _{1 is} a hydrogen atom.

The compounds according to the invention and their salts can be converted into medicaments with customary additives. These are administered in the usual way. The dose of the active ingredient is tuned in ÜR ^K! Icher way.

Corresponding pharmaceutically acceptable salts of the compounds of the general formula I are inorganic or organic salts such as hydrochlorides, sulfates, carbonates, phosphates, formates and maleates. Salts of the carboxylic acid of the general formula I in which R 1 denotes a hydrogen atom are, for example Sodium, potassium, calcium or ammonium salts and salts with organic amines. The salts are in made in the usual way.

The examples illustrate the invention.

example 1

A) Preparation of (±) -cis-7- [2- (2-tritylamino-4-thiazolyl) -2-anti-methoxyiminoacetamido] -2-tert-butoxycarbonyl-1-azabicyciol4,2,0] oct-2 -en-8-on according to the following reaction scheme:

Trityl -NH-

H
CCON

H ₃ CO-N

.y

CO ₂ tert-butyl

CO ₂ tert-butyl

Method a)

73 mg (0.307 mmol) of the compound obtained according to Example 4 and 135.9 mg (0.307 mmol) of 2- (2-tritylamino-4-thiazolyl) -2-anti-methoxyiminoacetic acid are dissolved in 2 ml of anhydrous methylene chloride and, while cooling and stirring, with a solution of 69.6 mg (0.307 mmol) of dicyclohexylcarbodiimide in 1 ml - Anhydrous methylene chloride added. The mixture is stirred for 3 hours. Then leave it 16 to Continue to react for 18 hours at OK ° C. Then the reaction mixture is diluted with an oil-based aqueous phosphate solution, saturated sodium bicarbonate solution and gcsäs.igwr sodium chloride solution. the Solution is dried over magnesium sulfate and evaporated under reduced pressure. 257 mg are obtained of a raw product. This is removed from a column charged with 12 g of silica gel with seed / a mixture η-hexane and ethyl acetate (1: 1) as eluents chromatographically purified. Yield 73 mg (35.9%) of the desired connection in vitreous form with pale yellow color.

C ₁ ;;

1780, 1725, 1695 (shoulder), 1690, 1635 NMR (CDCl,) <5 (ppm):

8.48 (d, 1H, J = 6.4 Hz); 7.25 (s, 15H); 6.30 (t, 1H, J = 3.0 Hz); 5.35 (t, 1H, J = 6 4 H /)
4.05 (s, 3H); 2.5-1.6 (m, 4H); 1.52 (s, 9 H)

Method b)

524.9 mg (1.18 mmol) of 2 - (? - tritylamino-4-lhiazolyl) -2-aiitimethoxyiminoacetic acid are dissolved in 10 ml of anhydrous tetrahydrofuran. The solution is mixed at -30 ⁰ C with 1.18 ml (1.18 mmol) of a solution of N-methylmorpholine in tetrahydrofuran and 1.18 ml (1.18 mmol) of a solution of isobutyl chloroformate in tetrahydrofuran. The mixture is stirred for 40 minutes. The solution is then treated with a mixture of 235 mg (0.987 mmol) of the compound (±) -cis-7-amino-2-part-butioxycarbonyl-1-azabicyclo [4.2.0] oct-2-en-8 -on poisoned dropwise in 5 ml of anhydrous ethylene chloride. It lets the Reaktionsaemisch 30 minutes to react and stirred for 2 hours at 0 ⁰ C. Then w - .. ro n 10 ml of ethyl acetate added. The reaction mixture is washed with water, saturated sodium bicarbonate solution and saturated sodium chloride solution. The solution is dried over sodium sulfate and evaporated under reduced pressure. 865 mg of a crude product are obtained. This is chromatographed on 40 g of silica gel according to method (a). Yield 580 mg (87.5%) of the desired compound. The! R and NMR data of the compound agree with the corresponding data of the compound fertilization. Jio is obtained according to method (a).

B) preparation of (±) -cis-7- [2- (2-amino-4-thiazolyI) -2-antimethoxyiiP ^; r! 0-acetamido] -l-azabicyclo [4,2.0] oct-2-en-8-one-2-carboxylic acid according to the following reaction scheme:

Tritvl-NH

CCONH

Il \

H ₃ CO-N

CO ₂ tert-buty!

CCONH

H ₃ CO-N jf]

O = Un J

CO ₂ H

500 mg (0.754 mmol) of the compound obtained above according to A) are in a mixture of 5 ml Trifluoroacetic acid, 2.5 ml of anhydrous methylene chloride and 2.5 m) anisole dissolved. The solution is left for 3 hours and stand for 40 minutes at 0 ° C and then stand under reduced pressure. The concentrate is made with 5 ml 50 percent aqueous acetic acid is added, then stirred at room temperature for 3 hours and under reduced pressure Pressure restricted. The concentrate is digested and nitrated well with diethyl ether. 244 mg of one are obtained Raw product. This is on a column loaded with 10 ml Diaion HP-IO using a mixture from methanol and water (2: 5 parts by volume; the same applies in the following examples) as Eluent purified by chromatography. Yield 90 mg (32.7%) of a pale yellow powder.

²⁰ IR (KBr) Cl _x ':

1760, 1670, 1630

MMR (CD-.OD) δ (ppm):

7.47 (s, 1H); 6.40 (ni, lH); 5.51 (d, 1H, J = 5.0 Hz); 4.05 (s, 3H); 4.3-3.7 (m, 1H); 2.6-1.1 (m, 4H)

Example 2

A) Preparation of (±) -cis-7 /; - [2- (2-Tritylamino-4-thiazolyl-2-niethoxyiminoacetamido] -4fl'-methyl-2-tert-butyloxycarbonyl-1-azabicyclo [4.2 , 0] oct-2-en-8-one according to the following reaction scheme:

H ₃ NHH

Trilvl -NH

CH ₃

H
CCON HH

H ₃ CON

CH ₃

CO ₂ tert-butyl

Method a)

88 mg (0.35 mmol) of the compound (±) -cis-7jö-amino-4ff-methyl-2-tcrt.-butyloxycarbonyl-1-azabicyclo- [4.2.0] oct-2-en-8-one dissolved in 1.5 ml of anhydrous methylene chloride and treated with 155 mg (0.35 mmol) of 2- (2-tritylamino-4-thiazolyl) -2-anti-methoxyiminoacetic acid. Furthermore, 1.5 ml of anhydrous dioxane are added to make the mixture more homogeneous. A solution of 80 mg (0.39 mmol) of dicyclohexyicarbodiimide in 1 ml of dioxane is then added. The mixture is stirred at 5 to 10 ^° C. for 16 to 18 hours. The white precipitate formed is filtered off. 10 ml of ethyl acetate and 5 ml of diethyl ether are added to the filtrate. The mixture is washed in sequence three times with 5 ml of 1 percent cold phosphoric acid and with saturated sodium bicarbonate solution. The solution is dried over sodium sulfate and evaporated under reduced pressure. 290 mg of the desired compound are obtained in crude form with a semi-solid consistency. The crude product is poured onto a column charged with 27 g of silica gel and treated with a mixture of nI lexane and ethyl acetate (2: eluted. The elual is evaporated under reduced pressure. Yield 170 mg (72%) of the desired compound with a melting point of 214.5 to 215.5 ⁰ C (umkrislallisiert from a mixture of η-hexane and ethyl acetate).

3240.1780 (shoulder). 1762, 1730, 1665, 1636

NNfR (CDCI., - CD, 0D) (5 (ppm):

7.30 (15 1/3 H, s); 6.64 (2/3 H, s); 6.34 (1H, dJ = 7 Hz); 5.48 (2/3 H, dJ = 5.5 Hz);

5.43 (1/3 H, dJ = 5.5 Hz); 4.08 (1H, s); 4.01 (2H, s); 3.90 (1H, m); 2.66 (1H, m); 1.75 (2H, m); 1.54 (3H, s); 1.53 (6H, s); 1.14 (3 H, d, J = 7 Hz)

Method b)

243.9 mg (0.05 mmol) of 2- (2-tritylamino-4-lhiazorylj-2-antimethoxyiminoacetic acid are dissolved in 5 ml of anhydrous tetrahydrofuran and mixed with 0.55 ml (0.55 mmol) of a 1 η solution of N- Methylmorphoün vcrsci / t. to the mixture "aräesj hei 0 ⁰ C was added dropwise with stirring 0.55 ml (0.55 mmol) of a solution of in Chlorameisensäureisobutyiester in tetrahydrofuran. Nacli lSminütigem stirring 0:11 ml (0.5 mmol) of triethylamine and then 144 mg (0.5 mmol) of the hydrochloride of ^ j-cis-T / f-amino ^ c-methyl ^ -tert-butyloxycarbonyl-1-azabicyclo [4.2.0] oct-2-en-8 -one were added. the mixture is stirred for 16 to 18 hours at 5 to 10 ⁰ C and then concentrated under reduced pressure. the concentrate is mixed with 10 ml of ethyl acetate and then sequentially with of 5 per cent hydrochloric acid, saturated sodium chloride solution, saturated sodium bicarbonate solution and saturated sodium chloride solution The solution is dried over sodium sulfate and evaporated under reduced pressure old crude product is placed on a column loaded with 25 g of silica gel and eluted with a mixture of η-hexane and ethyl acetate (5: 3). The eluate is evaporated under reduced pressure. Yield 250 mg (74%) of the desired compound. The properties of the compound agree with the properties of the compound obtained according to method (a).

B) Preparation of (±) -cis-7 /? - [2 (2-Amino-4-thiazolyl) -2-antimethoxyiminoacetamido] -4ff-methyl-2-carboxyl-l-azabicyclo- [4,2,0] oct-2-en-8-one according to the following reaction scheme:

Trityl -NH

H
CCON H

H ₃ CO-N

CO ₂ tert-butyl

CO ₂ H

70 mg (0.103 mmol) of the compound obtained above according to A) are dissolved in 0.5 ml of anhydrous methylene chloride and 0.1 ml of anisole. The mixture is ^{cooled to 0} ° C. and then 0.5 ml of trifluoroacetic acid is added. After standing 3½ hours in an ice bath, the reaction mixture is concentrated under reduced pressure. The concentrate is digested and nitrated with 5 ml of anhydrous diethyl ether, a white powder being obtained. This is dissolved in 2 ml of 50 percent acetic acid. Solution 2 ¹ / is allowed: hours at room temperature and then for 16 to 18 hours at 5 to 10 ⁰ C are. After weitcrem leaving the solution for 6 hours at 25 ⁰ C it is evaporated under reduced pressure to obtain a glassy product is obtained. This is digested well with diethyl ether and filtered. The filtrate is dried. Yield 20 mg (51%) of the desired compound.

IR (KBr) Cx ':

3480, 3300, 1770, 1680, 1635
NMR (DMSO-d ") <5 (ppm):

9.17 (1H, d, J = 8 Hz); 7.50 (1H, s); 7.24 (2H, m); 6.31 (1 H. d, J = 6 Hz); 5.52 (1H, m);

4.00 (3H, s); 2.65 (br, partially overlapping with the signal from DMSO-d ,,); 1.70 (2H, m);

1.06 (3 H, d, J = 7.5 Hz)

Example 3

A) Preparation of (±) -cis-7> [2- (2-tritylamino-4-thiazolyl) -2-methoxyiminoacetamido] -4 ^ -methyl-2-tert-butyloxycarbonyl-la / -abieyclo [4.2 , 0] oct-2-en-8-one according to the following reaction scheme:

Trityl -NH

H
CCON H

CH,

H ₃ CON

CO ₂ tert-butyl

CO ₂ tert-butyl

According to Example 2A), method (b), but using 202 mg (0.7 mmol) of the compound (±) -cis-7j8-amino-4yi-methyl-2-tert.-butyloxycarbonyl-1-azabicyclo [4 2.0] oct-2-en-8-one, are obtained 251 mg (53%) of the desired compound, melting at 201.0 to 202.0 ⁰ C.

(.0

3225, 1780 (shoulder), 1760, 1725, 1668, 1635

NMR (CDCh-CDiOD) δ (ppm):

7 35 U / 2 H, s); 7.30 (15H, s); 6.65 (1/2 H, s); 6.14 (1H, d, J = 2 Hz); 5.38 (1H, dd, J = 5 Hz);
4.09 (3/2H, s); 4.01 (3/2H, s); 3.96 (1H, m); 2.45 (1H, m); 2.05 (2H, m); 1.53 (9/2H, s);
1.52 (9/2H, s); 1.16 (3 H, dd, J = 7.5 Hz)

B) Preparation of (±) -cis-7jS- [2- (2-Amino-4-thiazolyl) -2-methoxyiminoacetamido] -4j? -Methyl-2-carboxyl-azabicyclo [4,2.0] oci-2 -en-8-on according to the following reaction scheme.

Trityl-NH

H CCON H

H ₃ CON

.CH ₃

H CCON H

H ₃ CON

CH ₃

CO ₂ tert-butyl

CO ₂ H

IO

15th

20th

3460, 3280, 1780 (shoulder), 1770, 1670, 1630

NMR (DMSO-d ") δ (ppm):

9.24 (1/2 H, d, J = 8 Hz); 9.17 (1/2 H, d, J = 8 Hz); 7.50 (1/2 H, s); 7.25 (2H, m); 6.78 (1/2 H, sj; 6.10 (1H, d, J = 2 Hz); 5.47 (1H, m); 4.00 (3/2 H, s); 3.85 (3/2H, s); 2.60 (br, partly with the signal from DMSO-d “overlapping); 1.91 (2H, m); 1.12 (3 H, d-d, J = 7.5 Hz)

Example 4

A) Preparation of (±) -cis-7je- [2- (2-chloroacetylamino-4-thiazolyl) -2-syn-methoxyiminoacetamido] -4amelhyl-2-carboxy-l-azabicyclo [4.2.0] oct- 2-cn-8-on according to the following reaction scheme:

According to Example 2B), but using 70 mg of the compound obtained above according to A) 22 mg (56%) of the desired compound are obtained:

ClCH ₂ CONH

40

50

55

60

65

H ₃ NHH

O =)

CH ₃

CF ₁ CO ₂ H

H
CCON H

H ₃ CON

CH ₃

CO ₂ H

CO ₂ H

172 mg. l, to prepare a homogeneous solution. Under stirring and cooling with a mixture of ice and salt, the reaction mixture with 129 mg (0.62 mmol) of phosphorus pentachloride is added. the mixture is stirred ^1] h hours and then 13.8 prepare ml η The supernatant liquid is decanted off and 1.3 ml of anhydrous tetrahydrofuran are added in order to obtain a solution of the acid chloride.

On the other hand, 160 mg (0.52 mmol!) Of the compound obtained in Reference Example 7 in 1 ml becomes 50 percent Dissolved aqueous tetrahydrofuran solution and treated with 209 mg (2.06 mmol) of triethylamine. This The mixture is added to the aforementioned acid chloride solution while cooling with ice and while stirring. To After stirring for 1 hour at the same temperature, the mixture is brought to pH 4 with 1 η hydrochloric acid set to 5 and extracted three times with 10 ml of ethyl acetate each time. The ethyl acetate extracts are combined with Washed sodium chloride solution, dried over sodium sulfate and evaporated under reduced pressure. Yield 147 mg (52.1%) of the desired compound.

IR (KBr) iC ':

1765, 1680

PMR (DMSO-d ") d (ppni):

7.40 (1H, s): 6.32 (1H, d, J = 5.2 Hz); 5.53 (1H, m); 4.35 (2H, s); 3.90 (3H, s); 2.50 (1H, m);
1.90-1.27 (2H, m); 1.10 (3 H, d, J = 7.5 Hz)

12th

B) Preparation of (±) -eis-7jff- [2- (2-Amino-4-thia / .olyl) -2-syn-mclhoxyiminoiicctamido] -4ir-mcthyll-azabicyclo [4,2.01oct-2-ene -8-one-2-carboxylic acid c according to the following scheme:

ClCH ₂ CONH

H ₂ N

H CCON H

H ₃ CON

CH ₃

N '\ H

CCON H H

H ₃ CON

CO ₂ H

CH ₃

CO ₂ H

147 mg (0.321 mmol) of the chloroacctyl compound obtained above according to A) are dissolved in 0.5 ml of dimethylsulfide and 2.5 ml of dimethylformamide and at room temperature with stirring with 47 mg (0.64 mmol) thiourea added. The mixture is stirred for 14 hours and then diethyl ether is added. the supernatant liquid is decanted off. The residue is in a small amount of dimethyl sulphide solved. The solution is applied to a column loaded with 10 ml of Diaion HP-IO. This comes with 240 ml Washed with water and eluted with a mixture of methanol and water (1:10 to 1: 2). The eluates are collected. The methanol is separated off under reduced pressure. The residue is back on a with Add 10 ml of Diaion HP-IO loaded column. This is washed with 500 ml of water. Elution occurs with a mixture of methanol and water (1: 1). The eluates are collected and reduced under reduced pressure Pressure evaporated. Yield 50.2 mg (41.1%) of the desired compound.

4.2 Hz); 5.58 (1H, br);

1760, 1680, 1655 PMR (DMSO-d ₆ ) <5 (ppm):

9.27 (1H, d, J = 9.0 Hz); 7.15 (2H, br); 6.75 (1H, s); 6.31 (1H, d, J 3.85 (3H, s); 2.60 (1H, m); 1.67 (2H, br); 1.08 (3 H, d, J = 8 Hz)

Example 5

A) Preparation of (±) -cis-7jS- [2- (2-chloroacetylamino-4-thiazoIyl) -2-syn-methoxyiminoacetamido] -l-azabicyclo [4,2.0] oct-2-en-8- on-2-carboxylic acid according to the following reaction scheme:

ClCH ₂ CONH

H ₂ NH

O = J

H CCON H

H ₃ CON

CO ₂ H

CO ₂ H

54.2 mg (0.195 mmol) of 2-chloroacetylamino-4-thiazolyl-2-syn-methoxyiminoacetic acid are dissolved in 0.98 ml Suspended anhydrous methylene chloride and mixed with 23.48 mg (0.195 mmol) of triethylamine. In addition 40.8 mg (0.195 mmol) of phosphorus pentachloride are added to the mixture while cooling with ice. After 20 minutes 3.92 ml of η-hexane are added while stirring. The supernatant liquid is decanted off. The residue is dissolved in 1.96 ml of tetrahydrofuran to obtain a solution of the acid chloride.

On the other hand, 45.9 mg (0.155 mmol) of the trifluoroacetate becomes the compound obtained in Referential Example 3 dissolved in 2 ml of 50 percent aqueous tetrahydrofuran solution and treated with 47.4 mg (0.469 mmol) of triethylamine offset. The above-mentioned acid chloride solution is added to this solution while cooling with ice. The mixture is adjusted to pH 2.0 with 10 percent hydrochloric acid and then extracted three times with ethyl acetate. The extracts are washed with saturated sodium chloride solution. The solution is over sodium sulfate dried and evaporated. Yield 80 mg of the desired compound in the form of a pale yellow Powder.

IR (KBr) r ;, ', " _IV ':

1760, 1710, 1660

PMR (DMSO-d ") <5 (ppm):

16.64 (1H, br); 9.39 (1H, d, J = 8.8 Hz); 7.47 (1H, s); 6.31 (1H, t): 5.51 (1H, d-d, J = 5.5; 8.8 Hz); 4.38 (2H, s); 3.89 (3H, s); 2.54-0.8 (4 H, m)

B) Preparation of (±) -cis-7- [2- (2-amino-4-thiazolyl) -2-syn-methoxyiminoacetatenido] -l-azabicyclo [4,2,0] -oct-2-en-8 -one-carboxylic acid according to the following reaction scheme:

5 ClCH ₂ CONH- ^

H ₃ N

H CCON H H

H ₃ CON

CCON H H H, C0N

CO, H

((UI

You-

80 mg of the compound obtained above according to A1 are dissolved in 0.% ml of nimelhylueeumiul
The solution is stirred at room temperature with 27.5 mg Tnioharrtsu> iT \ ersei / l wu! ilan »1! Siiüu! ·. · »Moved. After adding diethyl ether, the liquid overlooked becomes .ilulekaniieil. leaving a rolei nlip.ei residue. This is purified by chromatography on Diaion IIP-IO. Ausheule I ¹ U mg of the desired compound.

IR (KBr) v ™ v ':

1760, 1670, 1630

PMR (DMSO-d ") δ:

9.26 (1H, d, J = 8.6 Hz); 7.11 (2H, br); 6.75 (1H, s); 6.30 (1H, t); 5.47 (1H, d-d, J = 5.4, 8.8 Hz); 3.89 (3H, s); 2.5-1.0 (4 H, m)

The antibacterial effectiveness of the compounds obtained according to Examples 2 to 5 is according to the Heart infusion agar dilution method determined at pH 7.2. The results are in the summarized below table in which cefazolin is given for comparison.

Microorganisms Minimal inhibitory concentration, link • jg / ml link Cefazolin link according to link according to according to Example 3 according to Example 5 Example 2 50 Example 4 12.5 <0.05 Staphylococcus aureus 209-p 12.5 100 12.5 12.5 0.4 Staphylococcus aureus Smith 25th > 100 12.5 12.5 0.78 Staphylococcus epidermidis 25th 0.2 12.5 <0.05 1.56 Escherichia coli NIHJC-2 0.4 1.56 <0.05 <0.05 1.56 Escherichia coli Juhl 0.78 0.2 <0.05 <0.05 0.78 Klebsiella pneumoniae 8045 0.1 1.56 <0.05 <0.05 3.12 Klebsiella pneumoniae Y-60 1.56 6.25 <0.05 0.78 > 100 Serratia marcescens T-26 25th 3.12 1.56 <0.05 50 Serratia marcescens T-55 1.56 0.78 0.1 <0.05 12.5 Proteus mirabilis 1287 0.2 0.78 <0.05 <0.05 12.5 Proteus vulgaris 6897 <0.05 0.78 <0.05 <0.05 > 100 Proteus morganii K.Y4298 0.78 0.4 <0.05 <Q, Q5 25th Froteus reitgeri KY4289 0.1 > 100 £ 0.05 50 > 100 Pseudomonas aeruginosa 145 > 100 0.4 > 100 0.1 > 100 Pseudomonas putida F264 1.56 <0.05

The following reference examples explain the preparation of the starting compounds used in the examples.

Reference example 1

Preparation of (±) -cis-2-tert-butyloxycarbonyl-7-azido-1-azabicyclo [4.2.0] oct-2-en-8-one
(cis compound) of the following formula:
N ₃ HH

O =

COO-tert-butyl

In the following, the terms "eis" and "trans" refer to the 3- or 4- position of the 2-azetidinone ring or based on the 6- or 7-position of the 1-azabicyclol4,2,0] octane ring.

1) Preparation of 2- [4- (3-butenyl) -3-azido-2-oxoazetidin-1-yl | -2-diethylphosphonoacetate tert-butyl ester

447 mg (1.78 mmol) of tert-butyl-a-aminodiethylphosphonoacetate are used, which is an oily liquid and has the following data:

-i

10

IR (undiluted) ν ^ς _η ί

3400, 1735-1745, 1020-1060

NMR (CDCI,) δ (ppm):

4.20 (d-q, 4H); 3.83 (d, 1H, J = 20 Hz); 1.76 (br, 2H); 1.50 (s, 9H); 1.35 (t, 6H)

Masscnspcklrurn (m / e): \ s

268 (M ⁴ )

The compound is dissolved in 25 ml of anhydrous diethyl ether and treated with 164 mg (1.96 mmol) of 4-pentene-1-al offset. The solution is stirred for 1 hour at room temperature. Then 200 mg molecular sieve (4 A; Wako Junyaku Co., Ltd .; the same molecular sieve is also used in the following) and 150 mg of anhydrous Magnesium sulfate added. This mixture is stirred for 1 hour.

The reaction mixture is nitrated under reduced pressure. The filtrate is evaporated under reduced pressure, a pale yellow oily product remaining behind. Anhydrous benzene is added to this. The mixture is evaporated under reduced pressure to a pale yellow oily residue. The presence of a Schiff base in the product is demonstrated by NMR. The product is dissolved in 12.5 ml of cyclohexane and 12.5 ml of anhydrous benzene and 0.369 ml (2.66 mmol) of triethylamine and 200 mg of 4 A molecular sieve are added. Then add to the mixture at room temperature for IV; Hours of stirring and dropwise added a solution of 319 mg (2.66 mmol) of azidoacetyl chloride in 12.5 ml of cyclohexane. The reaction mixture is stirred for 30 minutes and then diluted with 10 ml of benzene. The solution is washed with 5 percent hydrochloric acid, saturated sodium bicarbonate solution, deionized water and saturated sodium chloride solution, dried over sodium sulfate and evaporated under reduced pressure. The remaining brown oily product is identified as the crude product of the desired compound. It is placed on a column charged with 45 g of silica gel and eluted with a mixture of η-hexane and allyl acetate (1: 2). Two isomers are obtained with respect to the 3- and the 4-position. Yield 345 mg of cis isomer, 58 mg of trans isomer. Overall yield 54.2%. ^J5

Cis isomcr:

IR (CHCl ₁ ) Cx ':

2120, 1775, 1770 (shoulder), 1750 1740 (shoulder), 1645

NMR (CDCl,) <5 (ppm):

6.13-6.33 (1H, m); 4.93-5.17 (2H, m): 4.50-4.93 (2H, m); 3.80-4.40 (5H, m); 1.93-2.17 (4H, m); 1.50 (9H, s); 1.33 (6 H, t)

Trans isomer:

IR (CHClOC _x ':

2120, 1780, 1755, 1750 (shoulder), 1650

NMR (CDCl ₃ ) δ (ppm):

5.43-6.20 (1H, m); 4.80-5.30 (2H, m); 3.75-4.75 (7H, m); 2.0-2.50 (4H, m); 1.5t / (9 H. d);

1.17 <6 II, rn)
2) Preparation of (±) -cis-2-tert-butyloxycarbonyl-7-azido-1-azabicyclo [4.2.0] oct-2-en-8-one

298 mg (0.716 mmol) of the compound (cis compound) obtained in (1) are dissolved in 8.5 ml of dioxane and 25 ml of deionized water, and 30 mg of osmium tetroxide are added. After stirring for 30 minutes, 496 mg (2.32 mmol) of powdered Natnumperjo dat are added to the black reaction mixture over the course of 20 minutes. The mixture is stirred for 1½ hours and then extracted three times with 50 ml of diethyl ether each time. The diethyl ether extracts are combined and washed with saturated sodium chloride solution, dried over sodium chloride and evaporated under reduced pressure. The dark brown oily product is applied to a column filled with 5 u silica gel and eluted with a mixture of benzene and ethyl acetate (1: 2). The fractions positive in the 2,4-dinitrophenylhydrazine reaction are collected and evaporated. 235 ml of an oily product are obtained, which is the cis-isomer of the aldehyde. The product is dissolved in 15 ml of anhydrous acetonitrile. The solution is mixed with 27.1 mg (0.563 mmol) SO percent sodium hydride at room temperature while stirring in nitrogen oxide. After stirring for 20 minutes, the mixture is added to 20 ml of 2 per / ml aqueous acetic acid and extracted four times with 50 ml of diethyl ether each time. The dietary ether extracts are combined, washed with saturated sodium chloride solution, dried over sodium sulfate and evaporated under reduced pressure. 180 mg of an oily product are obtained, which as a crude product of the

desired cis compound is identified. This product is applied to a column charged with 5 g of silica gel and eluted with a mixture of η-hexane and ethyl acetate (3.5: 1 parts by volume). Yield 91 mg (51%) of the desired compound as white crystals, melting at 64.5 to 65.5 ⁰ C.

2130, 1790, 1730, 1640

NMR (CDCl,) δ (ppm):

6.30 (1H, t, J = 4 Hz); 4.93 (1H, d, J = 5 Hz); 3.80 (1H, q); 1.6-2.6 (4H, m); 1.52 (9H, s)

Reference example 2

Preparation of (±) -cis-2-carboxy-7-azido-l-azabicyclo [4.2.0] oct-2-en-8-one (cis compound)

of the following formula:

Nj H H

55 mg (0.224 mMoUdergemäß Comparative Example 1, the compound obtained are dissolved in 2 ml of trifluoroacetic acid. The solution is allowed for 10 minutes at room temperature and then concentrated by evaporation under reduced 2> printing. The concentrate is mixed with benzene. The solution obtained is under reduced pressure Yield 51 mg (100%) of the desired carboxylic acid in the form of a yellow semi-solid.

IR (CHCI.,) V ™; ':

2120, 1770 (shoulder), 1760, 1715, 1635
NMR (CD ₃ OD) δ (ppm):

6.48 (1H, t, J = 4 Hz); 5.10 (1H, d, J = 5 Hz); 3.83 (1H, q); 1.1-2.5 (4 H, m)

Reference example 3

Preparation of (±) -cis-2-carboxy-7-amino-l-azabicyclo [4.2.0] oct-2-en-8-one of the following formula:

H ₂ N

O =!

CO ₂ H

91 mg of the compound obtained according to Reference Example 2 are dissolved in 6.5 ml of ethanol and added with 26 mg 10 percent palladium-on-carbon was added. The mixture is at room temperature and ambient pressure Stirred for 2 hours in a stream of hydrogen. After nitriding to remove the catalyst, it will The filtrate was concentrated under reduced pressure. The concentrate is dissolved in 10 ml of methanol and then with 26 mg 10 percent palladium-on-carbon was added. The mixture is at room temperature and ambient pressure Catalytically reduced for 3 hours and 50 minutes. Then, using Hyflo Super CeI as a filter aid filtered. The filtrate is evaporated under reduced pressure. Yield 88 mg (100%) of one semi-solid product. This is identified as the desired amino compound based on the following data.

60 IR (KBr)

3450, 2950, 1770, 1650

Reference example 4

Preparation of (±) -cis-7-amino-2-tert-butyloxycarbonyl-1-azabicycIo [4.2.0] oct-2-en-8-one
according to the following reaction scheme:

H ₂ NH

CO ₂ tert-butyl

CO ₂ tert-butyl

178 mg (0.67 mmol) of the compound obtained according to reference example 1, item 2, are dissolved in 10 ml of ethanol and treated with 25 mg of 10 percent palladium-on-carbon. The mixture is stirred at room temperature in a hydrogen stream for 50 minutes and then filtered to separate the catalyst. The filtrate is evaporated under reduced pressure. Yield 159.5 mg (100%) of the desired compound as a yellow oil. "

IRiCHCl ₃ ) C « ¹ :

1775, 1725, 1640

NMR (CDCl,) δ (ppm):

6.27 (m, 1H); 4.50 (m, 1H); 4.2-3.1 (m, 3H); 2.6-1.7 (m, 4H); 1.5 (s, 9 H)

If the catalytic reduction is carried out in ethanol, which contains an equivalent amount of hydrogen chloride, the hydrochloride of the aforementioned compound is obtained.

Reference example 5 U

Preparation of (±) -cis-2-tert-butyioxycarbonyl-4-methyl-7-azido-1-azabicyclo [4.2.0] oct-2-en-S-one

of the following formula:

H CH ₃ , ₀

1) Preparation of the tert-butyl ester of 2- [4- (2-methyl-3-butenyl) -3-azido-2-oxoazetidin-1-yl] -

2-diethylphosphonoacetic acid of the following formula:

CH ₃

CO ₂ tert-butyl

2.13 g (8 mmol) of tert-butyl-a-aminodiethylphosphonoacetate are dissolved in 80 ml of anhydrous diethyl ether dissolved and then treated with 902 mg (9.2 mmol) of 3-methyl-4-pentenal with stirring. The mixture is 1 hour stirred at room temperature. Then 900 mg molecular sieve 4 A and 700 mg magnesium sulfate are added. The mixture is stirred for 1½ hours and then filtered under reduced pressure. The filtrate will evaporated, leaving a pale yellow oil. This is mixed with 30 ml of anhydrous benzene. The -15 The mixture is evaporated to give 2.82 g of an oil. The presence of a SchifP's base detected. The oily product is dissolved in 56 ml of anhydrous cyclohexane and 56 ml of anhydrous Dissolved benzene and added 900 mg of molecular sieve 4 A and 1.67 ml (12 mmol) of triethylamine. then a solution of is added dropwise to the mixture with stirring at room temperature over a period of 1/2 hour 1.43 g (12 mmoles) of azidoacetyl chloride were added to 56 ml of anhydrous cyclohexane. After stirring for 30 minutes the reaction mixture is mixed with 30 ml of benzene in a separatory funnel. The Ben7 (ihchicht is with 30 ml each of 10 percent citric acid, saturated sodium chloride solution, saturated sodium bicarbonate solution and saturated sodium chloride solution, dried over sodium sulfate and under reduced Pressure evaporated. 2.8 g of an oil are obtained. By thin layer chromatography (silica gel, mixture from n-hexane and ethyl acetal (1: D) the presence of 2 isomers in the product is demonstrated. This will added to a column charged with 300 g of silica gel and with a mixture of η-hexane and ethyl acetate (1: 1) cluicrt. Yield 380 mg (11.0%) of the less polar isomer, 570 mg (16.7 "..) of the more polar isomer and 201 mg (5.8%) of a mixture of the two isomers. It is found that the more polar isomer represents the c's form of the desired compound.

Less polar isomer (trans compound);

2110, 1770, 1745

NMR (CDCl ₃ ) O (ppm):

5.40-6.10 (1H, m); 5.27-4.90 (2.5H, m); 4.68 (0.5H, ei ;; 4.23 (6H, nv): 2.60-1.77 (3H, m);
1.53 (9H, s); 1.37 (6H, t, J = 7.0 Hz); 1.10 (3 H, d, J = 6.0 Hz)

17th

More polar isomer (cis compound): ¹

2110, Π65, 1745

NiViR (CDCl ₅ ) i5 (ppm):

5.45-6.13 (. H, m); 4.83-5.20 (2.5H, m); 4.67 (0.5H, d); 3.97-4.45 (6H, m); 1.77-2.55 (3 II. M); 1.50 (9H, s); 1.33 (6H, t); 1.08 (3H, d)

2) Preparation of (±) -cis-7-azido-2-tert-butyloxycarbonyI-4-methyl-1-azabicyclo [4.2.0] oct-2-en-8-one

240 mg (0.56 mmol) of the compound (cis compound) obtained in (1) is dissolved in 6.6 ml of dioxane and 2 ml Dissolved deionized water and then mixed with 20 g of osmium tetroxide. After stirring for 10 minutes this will be black reaction mixture in 30 minutes with 390 mg (1.82 mmol) of powdered sodium periodate in small Portions staggered. The mixture is stirred for 40 minutes and then extracted three times with 30 ml of diethyl ether each time. The diethyl ether layers are combined, washed with saturated sodium chloride solution, over Dried sodium sulfate and evaporated. 230 mg of an oil are obtained. This is on one with 6 g of silica gel charged column and eluted with a mixture of benzene and ethyl acetate (1: 2). The one in the 2,4-dinitrophenylhydrazine reaction positive fractions are pooled and evaporated. 185 mg of an oil are obtained which is the aldehyde of the desired compound. The product is immediately anhydrous in 8 ml Dissolved acetonitrile and at room temperature with stirring in a stream of nitrogen with 21.6 mg (0.45 mmol) 50 percent sodium hydride added. After stirring for 30 minutes, the reaction mixture is poured into 15 ml Added 2 percent aqueous acetic acid and then extracted twice with 20 ml of diethyl ether each time. The diethyl ether layers are washed with saturated sodium chloride solution, dried over sodium sulfate and evaporated under reduced pressure. The remaining oil is loaded onto a 20 g of silica gel Given column and eluted with a mixture of η-hexane and ethyl acetate (3.5: 1 parts by volume). yield 70 mg (48.17 ») of the desired compound in the form of a colorless oil which crystallizes on standing.

IR (KBr) Cv ':

2110, 1784, 1715, 1623

NMR (CDCI,) δ (ppm):

6.30 (4/5 H, d, J = 5.1 Hz); 6.10 (1/5 H, d, J = 2.7 Hz); 4.98 (4/5 H, d, J = 5.0 Hz);

4.89 (1/5 H, d, J = 5.0 Hz); 3.60-3.90 (1H, m); 2.65 (1H, m); 1.70-1.80 (2H, m); 1.51 (9H, s);

1.20 (3/5 H, d, J = 8.0 Hz); 1.13 (12/5 H, d, J = 8.0 Hz)

From the NMR data it can be seen that the crystals obtained are a mixture of the 4 <z-methyl isomer and represent the 4_ / i-methyl isomer in a ratio of about 4: 1. The compounds can be determined by chromatography on silica gel using a mixture of η-hexane and ethyiaceta! (3: 1) as solvent be separated. The more polar isomer corresponds to the 4jß-methyl compound, the less polar isomer the 4a-methyl compound.

CH ₃

CO ₂ tert-butyl 4> CH, isomer

CO ₂ tert-butyl 4a-CH! Isomer

4./?-CH, -isomer:

F. from 84.0 to 86.5 C ⁰

2135, 1783, 1715, 1622

NMR (CDCl,) (5 (ppm):

6.13 (1H, i.e. J = 2.7 Hz); 4.90 (1H, d, J = 5.0 Hz); 3.93-3.73 (1H, m); 2.53 (1H, m);
2.16-1.75 (2H, m); 1.53 (9H, s); 1.20 (3 H, d, J = 6.0 Hz)

4ff-CH -, - isomer:

F. from 82.0 to 84.0 C ⁰

IR (KBr) Cv ":

2120, 1790, 1721, 1630

NMR (CDCI,) Ö (ppm):

6.33 (1H, d, J == 5.0 Hz); 5.00 (1H, d, J = 5.5 Hz); 3.89-3.68 (1H, m); 2.66 (1H, m);
1.82-1.57 (2H, m); 1.53 (9H, s); 1.12 (3 H, d, J = 7.0 Hz)

29 Il 787

Reference example 6

Preparation of (±) -cis-7 / (- amino-4ff-methyl-2-tert-butyIoxycarbonyl-1-azabicyclo [4,2,0] uct-2-en-8-one

according to the following reaction scheme:

CH ₃

H ₂ NH

CH ₃

CO ₂ tert-butyl

CO ₂ tert-butyl

255 mg (0.67 mmol) of the compound obtained according to reference example 5, item 2 (less polar isomer) are dissolved in 10 ml of ethanol, and 100 mg of 10 percent palladium-on-carbon are added. The mixture is catalytically hydrogenated 1 h hours Urd then filtered to remove the catalyst. The catalyst is washed with methanol. The filtrate and the washing solutions are combined and evaporated under reduced pressure. The pale yellow oil that remains is dissolved in 8 ml of ethyl acetate. The solution is extracted five times with 3 ml of 10 percent citric acid each time. The aqueous layer is adjusted to pH 6 to 7 with potassium carbonate to give a white suspension. This is extracted twice with 5 ml of ethyl acetate each time and washed with saturated sodium chloride solution. The solution is dried over sodium sulfate. Yield 177 mg (76.6%) of an oily product.

IR (CHCI ₃ ) C _x ':

3400, 1770, 1720, 1630

NMR (CDCI.,) Δ (ppm):

6.23 (1H, d, J = 5.0 Hz); 4.53 (1H, d, J = 5.8 Hz); 3.93-3.47 (1H, m); 2.56 (1 H, m) 1.92 (2H, br); 1.80-1.60 (2H, m); 1.50 (9H, s); 1.31 (5 H, d, J = 7.0 Hz)

If the catalytic hydrogenation is carried out with an equivalent amount of hydrochloric acid, the corresponding Obtain hydrochloride of the compound.

Reference example 7

Preparation of the trifluoroacetate of (±) -cis-7> amino-4ff-methyl-2-carboxy-1-azabicycIo- [4.2.0] oct-2-en-8-one according to the following reaction scheme:

H ₃ NH

CH ₃

O =! N

CO ₂ tert-butyl

CH ₃

■ CF ₃ CO ₂ H

CO ₂ H

196 mg (0.78 mmol) of the compound obtained according to Comparative Example 6 are dissolved in 4.2 ml of anhydrous dichloromelhan, and 1.8 ml of trilluoroacetic acid are added at room temperature while stirring. After 1 k hours, the mixture is concentrated under reduced pressure. The concentrate is azeotroped with anhydrous benzene. The oil obtained is digested with diethyl ether and nitrated. Yield 167 mg (69.3%) of the desired compound in the form of a powder.

IR (KBr) C ':

3460, 2980-2500, 1780, 1685, 1630 PMR (D _: O, with DDS as internal standard) <5 (ppm):

'6.77 (1H, d, J = 5.8 Hz); 5.00 (1H, d, J = 5.6 Hz); 4.10 (1H, m); 2.83 (1H, m) 1.86 (2H, m) 1.15 (3 H, d, J = 8.0Hz)

Test report

The superiority of compounds having a carbacephem nucleus with a hydrogen atom in the 3-position is demonstrated by the data in the tables below. These are comparative data on the antimicrobial effectiveness between acylated 3-H carbacephem compounds (these are prepared from the compounds of the invention) and the corresponding compounds with known carbacephem nuclei with optionally substituted methyl groups on the C ₃ carbon atom, as well as those compounds with conventional cephalosporin nuclei.

The antimicrobial activities of the compounds given in the tables below were determined by the heart infusion agar dilution method (pH 7.2).

10

15th

20th

25th

30th

•! 0

50 55 60 65

19th

Table I.

Comparison of the minimum inhibitory concentrations

H ₂ N

L ₂ H

H*) Table II CH j «) Ν — Ν 12.5 Microorganism CH ₁ S ^ f N «·) 0.4 Comparison of the minimum inhibitory concentrations V ' 0.05 25th > 100 CH ₅ 0.05 S. aureus 209P 0.05 3.1 0.1 St. pyogenes 0.05 6.3 0.2 E. coü NlH JC-2 0.01 6.3 K. pneumoniae SRC-I 0.02 6.3 P. mirabilis PR-4 0.05 12.5 P. vulgaris CN-339 Carbacephemous nucleus; *) Connection with 3-H I **) Connection with known core.

Microorganism

H, N

X = H

CONH

N o

OCH,

-N ^ y-H

X = CH, CONH

NO = JNJ-CH ₂ OAC

\ γ

^OCH 'CO ₁ H

Staphylocossus aureus 209-P 12.5 Staphylococcus aureus Smith 12.5 Staphylococcus epidermidis 12.5

Ash'richia coli NIHJC-2 <0.05

Escherichia coli Juhl < 0.05

Klebsieüa pneumoniae 8045 i 0.05

Klebsieila pneumoniae Y-60 i 0.05 Serratia marcescens T-26 0.78

Serratia marcescens T-55 s 0.05

Proteus mirabilis 1287 <0.05

Proteus vulgaris 6897 <0.05

Proteus morganii KY 4298 ^ 0.05

Proteus rettgeri KY 4289 s 0.05 Pseudomonas aeruginosa 145 50 Pseudomonas putida F 264 0.1

* l compound with 3-H carbacephemous nucleus; * ·) Connection with known core.

12.5

12.5

12.5

<0.05

<0.05

<0.05

<0.05

<0.05 <0.05 <0.05 <0.05 100 <0.05

50
0.2 0.1 0.05 0.1 1.56 0.4 0.2

<0.05 0.05 0.4 50
<0.05

20th

Table III

■ - Sftyl

Microorganism

R, = H

Η, Ν— <Il CONH CO _: H Ri = OH

O = J N J-CHv

CO ₁ H

Ccl'uwxim

E. coli NIHJC-2
E. coli Juhl
K. pneumoniae 8045
K. pneumoniae Y-60
S. marcescens T-26
S. marcescens T-55
P. mirabilis 1287
P. vulgaris 6897
P. vulgaris 51 255 *)
P. morganii 4298
P. rettgeri 4289
P. inconstans F 429
E. cloacae F 1510
E. aerogenic F1948
C. freundii F 1526
C. freundii S 1256 *)
S. typhimurium E-9
S. sonnei ATCC 9290

<0.01

<0.01

<0.01

<0.01

0.4 0.05 <0.0!

<0.01 0.02 0.02

<0.01 0.02 0.2

25 0.1 3.12 0.02 0.01

*) ./i- Lactamase producing strain.

Table IV

Comparison of the effectiveness against Pseudomonas MIC ( _: xg / ml)

<0.01

<0.01

<0.01

<0.01

0.4 0.05 <0.0! 0.02

<0.01 <0.01 <0.01 0. Oi ö.05 1.56 0.02 0.78 0.02

<0.01

0.1 0.05

<0.01 0.05 1.56 0.2 0.02 0.02 0.02 0.05

<0.01 0.1 0.4

25 0.2 6.25 0.1 0.02

VCONH ^Χ

OCH ₃

OH

Microorganism Q

CO ₃ H

present application

-N J-CHOAX X = H

CO ₂ H

O = I

N — N —N — JX CH, S— <? ^ N

CHjOAC "|

CH-.

(Ceftizoxime) (Cefotaxime!

(Cermenoximl

P. aeruginosa 0.78 6.25

! = 1
P. aeruginosa 3.12 50

145
P. aeruginosa 1.56

S 1252 *)

*) Cephalosporinase producing strain.

6.25 3.12 12.5 12.5 6.25 3.12

3.12 12.5 3.12

Pseudomonas aeruginosa is known to be one of the most important pathogenic microorganisms and a Main goal of current antibiotic chemotherapy. The table above shows that one of the compounds with the 3-H carbacephem core has a much higher activity against Pseudomonas than that

21

Compound with the known nucleus or cephalosporins with the same acyl group. The superiority of the Compound is also confirmed in vivo, see Table V.

The values for ED ₅₀ are determined by in vivo infection tests on mice. Mice are infected intraperitoneally with Pseudomonas aeruginosa F 199 with a cell suspension of 2.5 × 10 ″ cells / ml of pathogenic cells and 5% mucin. After 1.3 and 5 hours, various doses of the compounds to be tested are injected subcutaneously and the ED _{50 is} determined, which expresses the dose at which 50% of the test animals survive.

Table V

Chemotherapeutic activity against Pseudomonas

aeruginosa infection in mice

H, N

MHK m

OCH,

OK
\ / V / , J \ / \ Microorganism Y
COjH P. aeruginosa
F 199 157
(6.25) CO ₂ H
Ccflizoxime 472
(25

The 4-methoxy (R ₂ = OCH ₁ , an alkoxy radical), 4-acetoxy (R ₂ = OCOCH ₃ , an acyloxy radical) and 4-azido (R ₂ = Nj) carbacephem compounds of the present invention have another unexpectedly beneficial property as shown in Table VI below.

Table VI

H ₃ CON

CONH

/ \ y

CO ₂ H

Microorganism

Claimed connections

X - »-OCH., X = jü-OAc

X = JS-N,

C +)

CHjOAC

COjH

corresponding connections with known core

Staphylococcus aureus 209-P 6.25 3.13 3.13 25th Staphylococcus aureus Smith 6.25 3.13 3.13 50 Staphylococcus epidermidis 6.25 6.25 12.5 50 Pseudomonas aeruginosa l = 1 6.25 6.25 6.25 6.25

The compounds with the claimed core have higher activity against gram-positive bacteria than the chicken compounds with the same level of activity against Pseudomonas aeruginosa. In the professional world it is known that cephalosporins of the new generation with a narrow and broad untimicrobial spectrum against gram-negative bacilli an undesirable tendency towards reduced activity against gram-positive bacilli Have germs. Because the compounds with the claimed core have an increased antimicrobial activity against

22nd

have gram-positive bacteria (S. aureus and S. pidermidis) while maintaining the high effectiveness against gram-negative bacteria (P. aeruginosa), the claimed compounds have preferred properties as chemotherapeutic agents.

Another beneficial feature of the 3-H carbacephem compounds is that they are relatively higher Stability against yj-lactamase, which is the main mechanism of resistance to pathogenic microorganisms represents jö-lactam antibiotics; see Table VII.

Table VII

Comparison of the stability against _ # - lactamase
(Relative; degree of hydrolysis)

Eiuymqucllc H, N- ζ ~ |] CONH (/dblu.xim

* Y Ia

P. vulgaris F 715 <0.1 1.3

S. marcescens F 266 <0.1 1.1

C. freundii F 620 18.5 25.0

C. freundii S 1256 0.4 0.7

The compounds with halogen atoms as the radical R ₂ are valuable intermediates which can be converted into the 3-H carbacephem compounds with a hydroxyl group, a lower alkoxy group or a lower acyloxy group as the radical R ₂ .

The valuable properties of the 3-H carbacephem compounds according to the invention explained above were not known on the priority date of the application and are unexpected for experts.

Claims (1)

Patent claims:
1. Carbacephem derivatives of the general formula I. H ₂ N- in which A ₁ 1 is a hydrogen atom, an alkyl group having 1 to 6 carbon atoms, alkenyl of 2 to 6 carbon atoms, alkynyl having 2 to 6 carbon atoms or a cycloalkyl group having 3 to 6 carbon atoms, wherein said radicals are optionally substituted by a halogen atom, a carboxyl , Cyano or carbamoyl group or a lower alkoxycarbonyl radical having 1 to 4 carbon atoms are substituted, R-. a hydrogen atom, an alkyl radical with 1 to 5 carbon atoms or an acyloxy radical with 1 to 5 carbon atoms and R ₁ a hydrogen atom or a radical of the general formula