NO780835L - PROCEDURE FOR PREPARATION OF CEPHALOSPORINE DERIVATIVES - Google Patents

Description

"Analogy method for the preparation of

antibacterially active cephalosporin derivatives"

This invention relates to the production of new 7(3-[(2-amino-1,2-dioxb-ethyl)amino]acyl-cephalosporin derivatives which have the formula:

R means hydrogen, lower alkyl, phenyl-lower alkyl, diphenyl-lower alkyl, tri(lower alkyl)silyl, tri(lower alkyl)stannyl, trihaloethyl or a salt-forming ion.

means hydrogen, lower alkyl, saturated or unsaturated cycloalkyl, phenyl, phenyl-lower alkyl, substituted phenyl or certain heterocyclic groups.

R2 means hydrogen or methoxy. The R2 substituent is in the α-configuration as indicated by the broken line.

R₁ means hydrogen>lower alkyl, phenyl-lower alkyl or cycloalkyl.

Rg means hydrogen or -CR4R5~CN.

R 4 and R 4 are each hydrogen or lower alkyl.

X means hydrogen, lower r.alkanoyloxy, carbamoyloxy > lower alkoxy, lower alkylthio, certain heterothio groups,

When X is pyridinium or carbamoyl-substituted pyridinium, the compounds can be structurally represented by the formula:

where Z is hydrogen or carbamoyl.

The asterisks denote asymmetric carbon atoms.

The different groups denoted by the symbols have the meanings given below, and these definitions apply to the whole of this description.

The lower alkyl groups comprise linear or branched hydrocarbon groups containing 1 to 7 carbon atoms, preferably 1 to 4 carbon atoms, and especially 1 or 2 carbon atoms.

Examples of the groups referred to here are methyl, ethyl, propyl, isopropyl, butyl and t-butyl. The lower alkoxy and lower alkylthio groups comprise such lower alkyl groups bound to an oxygen or sulfur atom respectively, e.g. methoxy, ethoxy, propoxy, methylthio, ethylthio and propylthio. Phenyl-lower alkyl and diphenyl-lower alkyl groups include such lower alkyl groups attached to one or two phenyl rings, preferably benzyl, phenethyl and diphenylmethyl.

The saturated and unsaturated cycloalkyl groups are the alicyclic groups having up to 7 carbon atoms. and up to 2 double bonds in the ring, e.g. the cycloalkyl groups cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl, the cycloalkenyl groups with up to 7 carbon atoms with one double bond, cyclobutenyl, cyclopentenyl, cyclohexenyl and cycloheptenyl, and the cycloalkadienyl groups with up to 7 carbon atoms with two double bonds arranged in different positions, such as 1, 4-cyclohexadienyl which is particularly preferred.

The substituted phenyl groups contain one or two substituents selected from halogen (preferably chlorine or bromine), lower alkyl (preferably with 1 to 4 carbon atoms, especially methyl or ethyl), lower alkoxy (preferably with 1 to 4 carbon atoms, especially methoxy or ethoxy) and hydroxy, e.g. 2-, 3- or 4-chlorophenyl, 2-, 3- or 4-bromophenyl > 2-, 3- or 4-hydroxy-phenyl, 3,5-dichlorophenyl, 2-, 3- or 4-methylphenyl, 2 -, 3- or 4-ethoxyphenyl.

The salt-forming ions denoted by R are metal ions, e.g. aluminum, alkali metal ions such as sodium or potassium, alkaline earth metal ions such as calcium or magnesium, or an amine salt ion, a number of which are known for this purpose, e.g. phenyl-lower alkylamines such as dibenzylamine, N,N-dibenzylethylenediamine, lower alkylamines such as methylamine, triethylamine, and N-lower alkylpiperidines such as N-ethylpiperidine. Sodium and potassium are the preferred salt-forming ions.

The halogens are the four common halogens, of which chlorine

and bromine are preferred. As for the trihaloethyl group denoted by R, 2,2,2-trichloroethyl is preferred.

Trimethylsilyl is the preferred tri(lower alkyl)silyl group.

The heterocyclic groups denoted by R^ are thienyl,

furyl or pyridyl, ie 2-thienyl, 3-thienyl, 2-furyl, 3-furyl, 2-pyridyl>3-pyridyl or 4-pyridyl.

Lower alkanoyloxy- denotes a group of the formula -OCO-lower alkyl, preferably where the lower alkyl group is methyl. The lower alkanoyloxy groups have up to 7 carbon atoms,

and those having up to 4 carbon atoms are preferred, and acetyloxy

is particularly preferred.

The heterothio groups denoted by X are

where Rg is hydrogen or lower alkyl (preferably with 1 to 4 carbon atoms, especially methyl or ethyl). Rg is preferably hydrogen.

R'7 is carboxy (COOH) or C00~ ion<+>, -COO-lower alkyl,

-SO^H, -SC^-lower alkyl or cyano.

The new compounds of formula I are prepared according to the invention by acylation of a cephem compound with the formula

with an acid of the formula

or an activated derivative thereof such as an acid halide, an activated ester such as the nitrophenyl ester or dinitrophenyl ester, or a mixed anhydride, and/or in the presence of a coupling agent such as dicyclohexylcarbodiimide.

The compound of formula II is preferably in the form of an ester, ie R is a group which can be easily removed, such as diphenylmethyl, which is preferred, t-butyl and trimethylsilyl.

A preferred synthesis comprises that the acid of formula III is first reacted with chloroformic acid alkyl ester in the presence of a base such as triethylamine and then with the diphenylmethyl ester of the compound of formula II. The resulting ester is then hydrolyzed with •trifluoroacetic acid and anisole to form the free carboxyl group in the 4-position. A salt can be obtained from the acid by reaction with a base with the desired cation.

This turnover can e.g. performed by resolution

or suspending the acid in a neutral organic solvent such as chloroform, tetrahydrofuran, methylene chloride, dioxane, benzene

or similar, and addition, at a reduced temperature of approx. 0-5°C, of an equimolar amount of the compound of formula II. The reaction product is then isolated by usual methods, e.g. by concentrating or evaporating the solvent.

According to an alternative method, a compound of the formula

preferably in the form of a salt such as trifluoroacetic acid; the urea salt, dissolved or suspended in an organic solvent such as acetonitrile, methylene chloride, chloroform, dimethylformamide, tetrahydrofuran, dioxane, benzene or the like, and converted into an ester, e.g. by forming the trimethylsilyl ester by reaction with bis-(trimethylsilyl)acetamide. The product is then brought to

react with a compound of the formula

where hal means a halogen atom, preferably chlorine, in an organic solvent such as that mentioned above, at a reduced temperature, e.g. about. 0°C, and in the presence of a neutral acid scavenger such as propylene oxide, butylene oxide or the like.

The compounds of formula I wherein X is pyridinium or carbamoyl-substituted pyridinium can be prepared by reacting a compound of formula i wherein X is acetoxy with pyridine or carbamoyl-substituted pyridine in a polar solvent such as water, and in the presence of a catalyst such as an alkali metal thiocyanate by the process described in US Patent 3,792,047 and German Publication No. 2,234,280.

Compounds of formula I where X is a heterothio group can also be prepared by first preparing a compound of formula I where X is acetoxy and then reacting this product with a mercaptan of the formula

elizes an alkali metal (preferably sodium) salt of the formula by the methods described in US Patents 3,855,213, 3,890,309 and 3,892,737. The starting material with formula III where Rg is hydrogen is prepared from an α-amino acid ester with the formula where R-^ has the above meaning and Y is a group that can be easily removed, e.g. diphenylmethyl, nitroferryl, dinitrophenyl, t-butyl or trimethylsilyl, which is reacted with an oxalic acid derivative of the formula where Hal means halogen, preferably chlorine, and Z is lower alkyl, in the presence of a base such as triethylamine. This reaction gives an intermediate product with the formula Treatment of this intermediate product with an acid, e.g. trifluoroacetic acid and anisole, gives the free acid with the formula

Treatment of the product of formula IX with ammonia or an amine NHR^ and then acidification affords the acylating agent III. Activated derivatives thereof are produced by reaction with thionyl chloride, esterification agent, anhydride etc. in a regular way.

Alternatively, an α-amino acid ester of formula VII, preferably the diphenylmethyl ester, the nitroferryl ester or the dinitrphenyl ester, is reacted with an oxalyl halide such as oxalyl chloride to form a compound of the formula

where hal means halogen, preferably chlorine, and Y is one of the preceding ester groups such as diphenylmethyl, p-nitrophenyl or 2,4-dinitrophenyl. Reaction of this derivative with ammonia or an amine HNR^gives a product of the formula

When Y is nitrophenyl or dinitrophenyl, the intermediate of formula XI can be reacted with the compound of formula II.

When Y is diphenylmethyl in formula XI, it is preferred to react this intermediate with eri acid, e.g. hydrochloric acid in glacial acetic acid, to form a compound of the formula

which is then reacted with the compound of formula II, preferably in the form of its diphenylmethyl ester, the ester group then being removed as described.

The carboxylate salts of the compound of formula I are formed by reaction of the carboxyl group in the cephalosporanic acid moiety, i.e. R is hydrogen, with any of the salt-forming ions indicated above.

The starting material of formula III wherein Rg is different from hydrogen; is prepared from an α-amino acid ester with the formula

where R^ has the meaning given above and Y is a group which can be easily removed, e.g. diphenylmethyl, nitrophenyl, dinitrophenyl, t-butyl, trimethylsilyl or the like, which is reacted with an oxalyl halide such as oxalyl chloride. to form an intermediate of the formula where hal means halogen, preferably chlorine, in a solvent such as dioxane. This intermediate is then reacted with a compound of the formula

e.g. in an organic solvent such as those mentioned above in the presence of an organic base such as dimethylaniline at reduced temperature, e.g. about. -20°C, and treatment of this intermediate with an acid, e.g. trifluoroacetic acid and anisole, gives the free acid of formula III. By reacting the compound of formula X with an oxalyl halide such as oxalyl chloride in a solvent such as dioxane at elevated temperature, e.g. 60 to 70°C, a compound of formula V is formed.

Alternatively, the starting material of formula III can be prepared from the starting material of formula VII by reacting the latter with a compound of the formula

The carboxylate salts of the compound of formula I are formed by reaction of the carboxyl group in the cephalosporanic acid part,

i.e. R is hydrogen, with any of the salt-forming ions described above.

It will be seen that the compounds of formula I are optically active due to the presence of asymmetric carbon atoms denoted by the asterisks. By choosing the appropriate starting material, it is possible to obtain the compounds of formula I as a mixture of optically active isomers or isolated as a single isomer.

The various isomers as well as their mixtures can be prepared according to the invention.

Preferred compounds produced according to the invention are the acids and alkali metal salts of formula I (ie R is hydrogen, sodium or potassium) where X is acetoxy or heterothio, especially 1-methyl-1H-tetrazol-5-yl-thio, R is cyclohexadienyl ,

phenyl or a heterocyclic group selected from 2-thienyl, 3-thienyl, 2-furyl, 3-furyl, 2-pyridyl, 3-pyridyl and 4-pyridyl; R 2 is hydrogen or methoxy, especially hydrogen; and R 1 , R 1 and R 1 - are each hydrogen or lower alkyl, especially O-hydrogen, and especially the D-isomers thereof.

The most preferred compounds of formula I are the acids

and the alkali metal salts where R 1 is 2-thienyl or phenyl, especially 2-thienyl; R 1 , R 1 , R 4 and R 1 are each hydrogen; and X is heterothio,

especially

The new compounds of formula I have a broad spectrum of antibacterial activity against both gram-positive and gram-negative organisms such as Staphylococcus aureiis, Salmonella schottmuelleri, Psudomonas aeruginosa, Protéus rettgeri, Escherichia coli, Enterobacter hafniae, Enterobacter cloacae, Klebsiella pneumoniae, Serratia marcescehs, etc. They can be used as antibacterial agents for combating diseases caused by

organisms such as the above; and in general they can be used

in the same way as cephradine and other cephalosporins. E.g. can

a compound of formula I or a physiologically acceptable salt thereof is used in various animal species in an amount of approx. 1 to 100 mg/kg daily, orally or parenterally, in a single dose or two to four divided doses, to treat infections of bacterial origin, e.g. 5.0 mg/kg for mice.

Up to approx. 600 mg of a compound of formula I or

a physiologically acceptable salt thereof can be introduced into an oral dosage form such as a tablet; capsule or elixir or in an injectable form in a sterile aqueous vehicle, prepared according to ordinary pharmaceutical practice.

The following examples shall serve to further illustrate the invention. All temperatures are in °C.

Example 1

[ D- a-[( 2- ethoxy- 1, 2- dioxoethyl) amino]- 2- thiophenacetic acid di ifenyl methyl ester

3.2 g (20 mM) of 2-D-thienylglycine diphenyl methyl ester are dissolved in 50 ml of methylene chloride. 1 g of triethylamine is added, and 1.3 g (20 mM) oxalic acid chloride ethyl ester in 20 ml of methylene chloride is added dropwise at -20°. After 2 hours, the reaction solution is shaken with water, dried over sodium sulphate and the solvent is distilled off in a vacuum. The remaining syrup is the pure product, [D-α-[(2-ethoxy-1,2-dioxoethyl)amino]-2-thiophenacetic acid diphenylmethyl ester.

Example 2

D- a- [ (2- ethoxy-l, 2- dioxoethyl) amino]- 2- thiophenacetic acid

To 9.5 g of D-a-[(2-ethoxy-1,2-dioxoethyl)amino]-2-thiopheneacetic acid diphenyl methyl ester is added 50 ml of a mixture of trifluoroacetic acid and anisole (4:1) at -10°. The trifluoroacetic acid and the anisole are distilled off to give an oily residue which is taken up in 100 ml of saturated sodium bicarbonate solution. This is extracted twice with 20 ml of ether, and the aqueous phase is acidified with hydrochloric acid. Repeated extraction with ethyl acetate and evaporation of the ethyl acetate solution gives D-α-[(2-ethoxy-1,2-dioxoethyl)-amino]-2-thiophenacetic acid as a grayish syrup which does not crystallize, yield 6.5 g.

Example 3

D- a-[( 2- amino- 1, 2- dioxoethyl) amino]- 2- thiophenacetic acid

6.5 g of D-a-[(2-ethoxy-1,2-dioxoethyl)amino]-2-thio.phen-acetic acid are dissolved in 100 ml of 5N alcoholic ammonia solution and kept in a glass autoclave at 40-50° for 10 hours. The reaction mixture is evaporated, the residue is dissolved in water and acidified to give D-α-[(2-amino-1,2-dioxoethyl)amino]-2-thiophenacetic acid in the form of white crystals which are recrystallized from water, yield 4.8 g; sm.p. 173-175°.

Example 4

7- 3-[[ D-[( 2-amino- 1, 2- dioxoethyl) amino]- 2- thienylacetyl] amino]- 3-[[( 1- methyl- 1H- tetrazol- 5- yl) thio] methyl ]- 8- oxo- 5- thi- l- azabicyclo-[ 4. 2. 0] oct- 2- ene- 2- carboxylic acid- diphenyl methyl ester

1.2 g (5 mM) of D-a-[(2-amino-1,2-dioxoethyl)amino]-2-thiophenedacetic acid is dissolved in 50 ml of absolute tetrahydrofuran, and 0.5 ml of triethylamine is added. 0.55 g (6 mM) chloroformate ethyl ester in 10 ml tetrahydrofuran is added dropwise at -10°. After 30 minutes, this reaction solution is added dropwise to a solution of 2.5 g (5 mM) 7-amino-3-[[(1-methyl-1H-tetrazol-5-yl)thio]-methyl]cephalosporanic acid diphenylmethyl ester in 30 ml of tetrahydrofuran. The mixture is stirred for 3 hours at 5°. The reaction solution is filtered and the filtrate is evaporated to give a brown solid foam which is dissolved in 25 ml of methylene chloride and treated with charcoal. 200 ml of ether are added, after which 7-3-[[D-[(2-amino-1,2-dioxoethyl)amino]-2-thienylacetyl]amino]-3-[[(1-methyl-1H-tetrazol-5 -yi)thio]methyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid diphenyl methyl ester precipitates as a beige powder, yield 3.2 g, m.p. . 106° (split.).

Example 5

73~[[ D~[( 2- amino- 1, 2- dioxoethyl) amino]- 2- thienylacetyl]- amino]-3- [[( 1- methyl- 1H- tetrazol- 5- yl) thiojmethyl]- 8 - oxo-5-thia-l-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid

2.3 g of the product from example 4 is stirred for 10 minutes at 0° in a mixture of trifluoroacetic acid and anisole (4:1). After evaporation of the trifluoroacetic acid and the anisole in vacuum, ether is added to the remaining oil, which then solidifies. The solid is dissolved in 3N sodium bicarbonate solution, filtered and acidified.

with 2N hydrochloric acid to pH 3. The product, 70-[[D-[(2-amino-1,2-dioxoethyl)amino]-2-thienylacetyl]amino]- 3-[[(1-methyl-1H-tetrazol- 5-yl)typ]methyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, precipitates as a brown powder which is dried, dissolved in tetrahydrofuran, treated with charcoal , and the acid is precipitated with ether as a light beige powder, yield 0.78 g, m.p. 158-16i°.

Example 6

D- a-[( 2- amino- 1, 2- dioxoethyl) amino]- 2- thiophene acetyl chloride

2.3 g of O-α-[(2-amino-1,2-dioxoethyl)amino]-2-thiopheneacetic acid are suspended in 80 ml of acetonitrile, and 1.2 g of thionyl chloride are added all at once at -20°. The temperature is allowed to rise to room temperature, and the solvent is distilled off in a vacuum. The sticky brownish residue is treated with ether to give a light beige solid powder. The infrared spectrum shows the desired acid chloride which is used immediately since decomposition takes place during storage.

Example 7

73~[[ D-[( 2- amino- 1, 2- dioxoethyl) amino]- 2- thienylacetyl] amino]-3-[[( 1- methyl- 1H- tetrazol- 5- yl) thio] methyl]- 8- oxo- 5- thia- l- azabicyclo-[ 4. 2. 0] oct- 2- en- 2- carboxylic acid

2.2 g (7.5 mM) of D-α-[(2-amino-1,2-dioxoethyl)amino]-2-thiopheneacetyl chloride are added at -10° to a solution of 2.5 g (7.5 mM) 7 -amino-3-f[(1-methyl-1H-tetrazol-5-yl)thio]methyl]-cephalosporanic acid and 3.7 g of bis-trimethylsilyl acetamide in 100 ml of acetonitrile. The mixture is stirred for 1 hour, and the solvent is then distilled off. The residue is taken up in 50 ml of methanol and 1 ml of 2N hydrochloric acid and treated with charcoal; The crude product 7f3-[ [D-[(2-amino-1,2-dioxoethyl)amino]-2-thienylacetyl]amino]-3-[[(1-methyl-1H-tetrazol-5-yl)thio]methyl] -8-oxo-5-thia-1-azabicyclo[4.2.0]-oct-2-ene-2-carboxylic acid is obtained as a beige powder which is reprecipitated from tetrahydrofuran/ether, m.p. 156-158°, and is identical to the product according to example 5.

Example 8

73- - 8-oxo-5-thia-l-azabicyclo [ 4. 2 . 0] oct- 2- ene- 2- carboxylic acid potassium salt

The product from Example 5 is reacted with an equimolar aqueous solution of potassium bicarbonate to give 73~t[D-[(2-amino-1,2-dioxoethyl)amino]-2-thienylacetyl]amino]-3-[[(1 -methyl-1H-tetrazol-5-yl)thio]methyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid potassium salt as a brownish powder,

sm.p. 174-175°.

Example 9

73~ amino- 3-[[( l- methyl- lH- tetrazol- 5- yl) thio] methyl]- 8- oxo-5- thia- l- azabicyclo[ 4. 2. 0] oct- 2- en- 2- carboxylic acid

To a stirred suspension of 27.2 g of 7-aminocephalosporanic acid (0.1 mol) in 150 ml of acetone and 100 ml of H20 at 0-5°, add 50 ml

2N NaOH, being careful to keep the pH below 8.5. A solution of 12.7 g (0.11 mol) of 1-methyl-5-mercapto-1H-tetrazole in 50 ml of 2N NaOH is added and the mixture is warmed to room temperature. The stirred mixture is then kept at 60° (internal temperature) under nitrogen for 3 hours at pH 7-7.5 with periodic addition of dilute aqueous NaOK. The mixture is cooled in an ice-water bath, and with stirring, 3N HCl is added to adjust the pH to 3.9. Stirring is continued for 15 minutes, and the precipitate is collected by filtration, washed with water and then acetone and finally dried to give the desired product as a powder (18.4 g).

Example 10

73- amino- 3-[[( l- methyl- lH- tetrazol- 5- yj) thio] methyl]- 8- oxo- 5- thia-1- azabicyclo[ 4. 2. 0] oct- 2- en- 2- carboxylic acid diphenyl methyl ester

A mixture of 16.4 g (0.05 mol) of the acid product of Example 9, 10.3 g (0.054 mol) p-toluenesulfonic acid monohydrate, 350 ml dioxane (dried by passing through basic alumina) and dry CH 2 OH are stirred at room temperature under nitrogen for 30 minutes. The clear solution is evaporated to a residue,

and water and CH^OH are removed by four evaporations of 100 ml of dioxane. Fresh dioxane (300 ml) is then added to the residue, followed by

solution of crystalline diphenyldiazoethane (19.4 g, 0.10 mol)

in 150 ml of dry dimethoxyethane. The mixture is first shaken vigorously for 10-15 minutes and then stirred at room temperature for 3 hours. Methanol (25 ml) is added, and the red solution therein is stirred until complete

it has turned yellow-orange. The solvents are removed in vacuo,

and the residue is treated with 400 ml of Cf^C^ and a solution of 20 g of K^WPO^ in 250 ml of water. The CH^^ layer is washed with water and saturated NaCl and finally dried (MgSO^) to give a residue after removal of the solvent in vacuo. Treatment of the residue with Et 2 O gives a solid (2 7 g). Bubble chromatography of this solid on silica gel eluting with CHCl3 and then EtOAc-CHCl3 (4:1) gives the desired product as a residue (12.9 g). Treatment with EtOAc then gives 8.0 g of the desired product

as a pale yellow powder.

Example 11

DL-g-[(2-ethoxy-1,2-dioxoethyl)amino]phenylacetic acid diphenylmethyl ester By using DL-α-aminobenzeneacetic acid diphenylmethyl ester instead of 2-D-thienylgycine diphenylmethyl ester in the method according to Example 1, DL-α-[(ethoxy-1,2-dioxoethyl)amino]-phenylacetic acid diphenylmethyl ester is obtained as a thick, colorless oil. Example 12 DL-α-[(2-amino-1,2-dioxoethyl)amino]phenylacetic acid diphenylmethyl ester A mixture of 10 mM DL-α-[(2-ethoxy-1,2-dioxoethyl)amino]phenylacetic acid diphenylmethylester in 50 ml of ethanol containing 13 mM ammonia is stirred for 15 minutes. After a short time a thick slurry forms. The product, DL-α-[(2-amino-1,2-dioxoethyl)-amino]phenylacetic acid diphenylmethyl ester is filtered off and recrystallized from toluene in the form of white filaments, m.p. 168°.

Example 13

DL-g-[(2-amino-1,2-dioxoethyl)amino]phenylacetic acid

13 g of the product from example 12 is added to 250 ml of

a 6N solution of hydrochloric acid in glacial acetic acid. After stirring in

15 minutes everything is dissolved. The reaction solution is evaporated to room temperature, and the white, crystalline residue is triturated with ether, filtered under suction and recrystallized from water to

give DL-α-[(2-amino-1,2-dioxoethyl)amino]phenylacetic acid as white crystals, m.p. 193°.

Example 14

73- oxo- 5- thia- l- azabicyclo-[ 4. 2. 0] oct- 2- en- 2- carboxylic acid- diphenyl methyl ester

i.85 g (7.5 mM) DL-g<->[(2-amino-1,2-dioxoethyl)amino]-phenylacetic acid is reacted with chloroformic acid ethyl ester and 7-amino-3-[[(1-methyl -1H-tetrazol-5-yl)thio]methyl]cephalosporanic acid diphenylmethyl ester by the method of Example 4 to give the product, 73-[[DL-[(2-amino-1,2-dioxoethyl)amino]phenylacetyl]amino] -3-[[(1-methyl-1H-tetrazol-5-yl)thio]methyl]-8-oxo-5-thia-1-azabicyclo-[4.2.0]oct-2-ene-2-carboxylic acid- diphenyl methyl ester, as a beige powder, m.p. 146°.

Example 15

73~[[ DL-[( 2- amino- 1, 2- dioxoethyl) amino] phenylacetyl] amino]- 3-[[( 1- methyl- 1H- tetrazol- 5- yl) thiojmethyl]- 8- oxo- 5 - thia-l-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid

By treating the product from example 14 with trifluoroacetic acid and anisole by the method according to example 5, 73-[[DL-[(2-amino-1,2-dioxoethyl)amino]phenylacetyl]amino]-3-[[(l- methyl-1H-tetrazol-5-yl)thio]methyl]-8-oxo-5-thia-1-azabicyclo-[4.2.0]oct-2-ene-2-carboxylic acid as a beige powder,

sm.p. 153° (decomposition).

The sodium salt is obtained as a light, beige powder by freeze-drying an equimolar, aqueous solution of the above-mentioned acid and sodium bicarbonate, m.p. 175° (split.).

Example 16

DL- g-[( ethoxy- 1, 2- dioxoethyl) amino]- 2- furanoacetic acid- diphenyl methyl ester

By using 2-DL-furylglycine diphenyl methyl ester

instead of 2-D-thienylgycine diphenyl methyl ester in the method according to example 1, DL-g<->[(ethoxy-1,2-dioxoethyl)amino]-2-furanacetic acid diphenyl methyl ester is obtained as a colorless oil which does not crystallize.

Example 17

DL- g-[( 2- amino- 1, 2- dioxoethyl) amino]- 2- furanoacetic acid- diphenyl methyl ester

By using DL-α<->[(ethoxy-1,2-dioxoethyl)amino]-2-furanacetic acid diphenylmethyl ester in the method according to example 12, DL-α-[(2-amino-1,2-dioxoethyl) amino]-2-tetraacetic acid diphenylmethyl ester as white crystals, m.p. 168-170° (toluene).

Example 18

DL- a-[( 2- amino- 1, 2- dioxoethyl) amino]- 2- furanoacetic acid

By using DL-α-[(2-amino-1,2-dioxoethyl)amino]-2-furoacetic acid diphenyl methyl ester in the method according to example 13, DL-α-[(2-amino-1,2- dioxoethyl)amino]-2-furanacetic acid as white crystals, m.p. 174° (water).

Example 19

( 6R- trans )- 7-[[ DL-[( 2- amino- 1, 2- dioxbetyl) amino]- 2- furanyl-acetyl] amino]- 3-[[( l- methyl- 1H- tetrazole- 5 - yl) thio] methyl]- 8-bxo- 5- thia- l- azabicyclo[ 4. 2. 0] oct- 2- ene- 2- carboxylic acid diphenyl methyl ester

By using DL-α-[(2-amino-1,2-dioxoethyl)amino]-2-furanacetic acid instead of D-α-[(2-amino-1,2-dioxoethyl)amino]-2-thiopheneacetic acid in the method according to example 4, (6R-trans)-7-[[DL-[(2-amino-1,2-dioxoethyl)amino]-2-furanyl-acetyl]amino]-3-[[(methyl-1H -tetrazol-5-yl)thiol-methyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid diphenyl methyl ester as a beige powder, m.p. 81° (cleavage).

Example 20

( 6R- trans )- 7-[[ DL-[( 2- amino- 1, 2- dioxoethyl) amino]- 2- furanyl-acetyl] amino]- 3-[[( 1- methyl- 1H- tetrazole- 5 -yl)thio]methyl]-8-oxo-5-thia-l-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid. By using the diphenylmethyl ester obtained in example 19 by the method according to example 5, you get mari (6R-trans)-7-[[DL-[(2-amino-1,2-dioxoethyl)amino]-2-furariylacetyl]amino]-3- [[(1-methyl-1H-tetrazol-5- yl)thio]methyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid as a beige powder, m.p. 154° (decomposition).

The sodium salt is obtained as a light powder by the method according to example 15, m.p. 171° (decomposition).

Example 21

L- a-[( 2- amino- 1, 2- dioxoethylj amino] thiophenacetic acid- diphenyl methyl ester

a) 3.5 g (10 mM) g<->thienyl gylcine diphenyl methyl ester hydrochloride is suspended in 50 ml of a mixture of carbon tetrachloride

and methylene chloride (2:1). 1.2 g (10 mM) oxalyl chloride is added. The mixture is stirred, and a stream of nitrogen is passed through the wood

35° until a clear solution is obtained. The greenish reaction solution containing L-[(2-chloro-1,2-dioxoethyl)amino]-thiophenacetic acid diphenylmethyl ester is decolorized with activated charcoal and used directly in the next step. b) The reaction solution from part a) is added dropwise to a cooled solution (-20°) of 10 mM ammonia and 10 mM dimethylaniline in 500 ml methylene chloride. After the addition, the reaction mixture is stirred for a further 15 minutes, washed with 100 ml of water, 100 ml of 2N hydrochloric acid and again with 100 ml of water, dried and evaporated. The residue is recrystallized from ethanol and then from toluene to give the product, L-g-[(2-amino-1,2-di'oxoethyl)amino]-thiophenacetic acid diphenylmethyl ester as white crystals,

sm.p. 157-160° [a]p° = +59.0° (1% in methylene chloride).

Example 22

L-g-[(2-amino-1,2-dioxoethyl)amino]-2-thiophenedacetic acid L-g<->[(2-amino-1,2-dioxoethyl)amino]-2-thiophenedacetic acid is obtained as white crystals on treatment of the diphenylmethyl ester obtained in example 21 with ammonia by the method according to example 12, m.p. 140-141° [ct]^° = +152° (1% in THF).

Example 2 3

73~[[ L-[( 2- amino- 1, 2- dioxoethyl) amino]- 2- thienylacetyl] amino]-3-[[( 1- methyl- 1H- tetrazol- 5- yl) thio] methyl]- 8- oxo- l- azabicyclo-[ 4. 2. 0] oct- 2- ene- 2- carboxylic acid

By using L-g<->[(2-amino-1,2-dioxoethyl)amino]-2-thiophenacetic acid by the method according to example 4 and recrystallization from isopropanol, one obtains 7j3- [ [L- [ (2-amino -1, 2-dioxoethyl)amino]-2-thienylacetyl]amino]-3-f[(1-methyl-1H-tetrazol-5-yl)thio]methyl]-8-oxo-1-azabicyclo[4.2.0 ]oct-2-ene-2-carboxylic acid diphenyl methyl ester as a beige powder, m.p. 94° (cleavage).

By treating this product as in the method according to example 5, 70-[[L-[(2-amino-1,2-dioxoethyl)amino]-2-thienyiacetyl]amino]-3-[[(1-methyl -1H-tetrazol-5-yl)thio]methyl]-8-oxo-1-azabicyclo[4.2.O]oct-2-ene-2-carboxylic acid, m.p. 150°

(fission). The sodium salt is obtained by the method according to example 15, m.p. 167-170°C (decomposition).

Examples 24-75

By following the procedure according to example 7, but by

to use the below indicated acylating agent A with the substituents in the following table (prepared as described in examples 1-3 and 6) and 70-aminocephalosporanic acid compound B below, one obtains the product C which has the substituents indicated in the table. When necessary, the protecting group and the ester group are removed as in examples 5 or 7. The salts are prepared as in example 8.

The acylating agents A can be in either D or L form

or may be a mixture of D and L isomers.

. Example 76

[(cyanomethyl) amino] oxoacetyl chloride

20 mM cyanomethylamiric hydrochloride and 24 mM oxalyl chloride

in 100 mM absolute dioxane is heated at 60-70° while a stream of nitrogen is passed through the mixture. After 1 hour, a clear solution is obtained, and there is no sign of hydrogen chloride in the nitrogen flow. The solvent is evaporated under vacuum, and the remaining light brown oil is taken up in methylene chloride, filtered over charcoal and stored at -30° until it is to be used.

Example 77

D- a-[[[( cyanomethyl) amino] oxoacetyl] amino]- 2- thiophenacetic acid diphenyl methyl ester

10 mM 2-D-thienylglycine diphenyl methyl ester is dissolved in 20 ml methylene chloride, 10 mM dimethylaniline is added, and half of the product from example 76 is added dropwise with stirring at -20°. After 30 minutes, the reaction solution is first washed with 50 ml

IN hydrochloric acid<p>g then with 50 ml of water. After drying over sodium sulphate, it is concentrated by evaporation. A viscous, yellow mass is obtained which hardens when treated with ether. It is crystallized from a little ethanol to give D-α-[[[(cyanomethyl)amino]oxoacetyl]amino]-2-thiopheneacetic acid diphenylmethyl ester as white crystals, yield 63%, m.p. 160-161°.

Example 78

D- a-[[[( cyanomethyl) amino] oxoacetyl] amino]- 2- thiophenacetic acid

By treating the diphenylmethyl ester obtained in example 2 with trifluoroacetic acid and anisole (4:1), D-a-[[[(cyanomethyl)-amino]oxoacetyl]amino]-2-thiopheneacetic acid is obtained in 74% yield. The product is recrystallized from absolute ethanol to give white crystals, m.p. 199-200°.

Example 79

73-[[ D-[[ 2-[( cyanomethyl) aminoj- 1> 2- dioxoethyl] amino]- 2- thienylacetyl ] amino]- 3-[[( l- methyl- lH^ tetrazbl- 5- y1) thio ] methyl]- 8- oxo-5- thia- l- azabicyclo[ 4. 2. 0] oct- 2- ene- 2- carboxylic acid 10 mM D-a-[[[(cyanomethyl)amino]oxoacetyl]amino]-2- thiophenacetic acid and 10 mM triethylamine are dissolved in 100 ml of tetrahydrofuran; is cooled to -5°, and a solution of 11 mM chloroformic acid ethyl ester is slowly added dropwise. The mixture is allowed to react for 20 minutes, and then a solution of 10 mM 7-amino-2-[[(1-methyl-1H-tetrazol-5-yl)thio]methyl]cephalosporanic acid diphenylmethyl ester in tetrahydrofuran is added. After stirring for 8 hours at -5° and 1 hour at room temperature, the solution is concentrated under vacuum and then taken up with methylene chloride and water. The organic phase is washed with 2N phosphoric acid solution and water, dried, treated with charcoal and concentrated. The product, 73-[[D-[[2-[(cyanomethyl)amino]-1,2-dioxoethyl]amino]-2-thienylacetyl]amino]-3-[[(1-methyl-1H-tetra zol-5 -yl)thio]methyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid diphenyl methyl ester is obtained as a solid, brownish foam. By treating this with trifluoroacetic acid and anisyl (4:1), mari obtains 7-3~[[D-[[2-[(cyanomethyl)amino]-1,2-dioxoethyl]amino]-2-thienylacetyl]-amino] -3-[[(1-methyl-1H-tetrazol-5-yl)thio]methyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid in the form of a beige powder that is recrystallized from isopropanol/ethyl acetate,

sm.p. 154°.

Example 80

Alternative synthesis of 7~ 3~[[ D~ l [ 2 -[( cyanomethyl) amino]- 1, 2- dioxoethyl] amino]- 2- thienylacetyl] amino]- 3-[[( l- methyl- lH- tetrazole - 5- yl)-thio] methyl]- 8- oxo- 5- thia- l- azabicyclo[ 4. 2. 0] oct- 2- ene- 2- carboxylic acid

10 mM 73-amino^3-[[(1-methyl-1H-tetrazol-5-yl)thio]-methyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene -2-carboxylic acid trifluoroacetic acid salt is suspended in acetonitrile. After adding 3 ml of bis(trimethylsilyl)acetamide and 15 mM propylene oxide, a clear solution is obtained within a few minutes. This is cooled to 0°, and 12 mM chlorooxoacetic acid-cyanomethylamide dissolved in methylene chloride is added dropwise. The mixture is stirred for 1 hour, and then the solvent is removed in vacuo. The residue is stirred for 1 hour in 300 ml of ethyl acetate and 50 ml of water. The organic phase is then dried, treated with charcoal and concentrated to 30 ml, whereby the product 7-3-[[D-[[2-[(cyanomethyl)amino]-1,2-dioxoethyl]amino]-2-thienylacetyl]amino ]-3- [ [(1-methyl-1H-tetrazol-5-yl)thio]methyl-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid crystallises. By pouring the mother luteh into ether, a further quantity of the product is obtained, m.p. 153-155°.

Example 81

73~[[ D-[[ 2-[( cyanomethyl) amino]- 1, 2- dioxoethyl] amino]- 2- thienyl-acetyl] amino]- 3-[[( l- methyl- lH- tetrazole- 5 - yl) thio] methylj- 8- oxo-5- thia- l- azabicyclo[ 4. 2. 0] oct- 2- en- 2- carboxylic acid sodium salt

By freeze-drying an equimolar aqueous solution of the acid from example 79 or example 80 and sodium bicarbonate, 73~[[D-[[2-[(cyanomethyl)amino]-1,2-dioxoethyl]amino]-2-thienylacetyl] is obtained amino]-3-[[(1-methyl-1H-tetrazoi-5-yl)thio]methyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid -sodium salt as a pale yellow powder, m.p. 178° (splint.).

Examples 82-145

By following the procedure according to example 79, but by using the below indicated acylating agent A which has the substituents in the following table (prepared as described in examples 76 to 78) and the 73-aminocephalosporanic acid compound B below, one obtains the product C which has those in the table indicated substituents. When necessary, the protecting group and the ester group are removed as in Example 79. The salts are prepared as in Example 81.

The acylating agents A can be in either D or L form or can be a mixture of D and L isomers.

Claims (1)

Analogy method for the preparation of an anti bacterially active compound with the formula where R is hydrogen, lower alkyl, phenyl-lower alkyl, diphenyl- lower alkyl, tri(lower alkyl)silyl, tri(lower alkyl)stannyi, trihaloethyl or a salt-forming ion; R^ is hydrogen, lower alkyl, saturated or unsaturated cycloalkyl, phenyl, phenyl-lower alkyl, substituted phenyl wherein the phenyl substituent is one or two residues selected from halogen, lower alkyl, lower alkoxy and hydroxy, or a heterbicyclic group selected from 2 -thienyl, 3-thienyl, 2-furyl, 3-furyl, 2-pyridyl, 3-pyridyl and 4-pyridyl; R 2 is hydrogen or methoxy; R 3 is hydrogen, lower alkyl, phenyl-lower alkyl or cycloalkyl; Rg is hydrogen or -CR4 R5~ CN; R4 and R5 are each hydrogen or lower alkyl,1 and X is hydrogen, lower alkanoyloxy, carbamoyloxy, lower alkoxy, lower alkylthio, or a heterothio group selected from Rg is hydrogen or lower alkyl, and R-, is carboxy, C00 ion+, -COO-lower alkyl, SO^ H, -S02 -lower alkyl or cyano, characterized by ata) a compound with the formula is acylated with a compound of the formula or its chemical equivalent, or b) a compound of the formula is reacted with a compound of the formula where hai means halogen, and the other substituents have the meanings indicated above, and eventually, for the preparation of a compound of formula I where X is pyridinium or carbamoyl-substituted pyridinium, a compound of formula I where X is acetoxy is reacted with pyridine or carbamoyl-substituted pyridine in a polar solvent and in the presence of a catalyst, or for the preparation of a compound where X is a heterothio group, a compound of formula I where X is acetoxy is reacted with a mercaptan of the formula or an alkali metal salt of the formula wherein hetero-S is a heterothio group as defined above.