SE437025B - 1- (2-SULFAMINOETHYL) -TETRAZOL-5-THIOL USED FOR THE PREPARATION OF CEPHALOSPORINE COMPOUNDS WITH ANTIBACTERIAL ACTIVITY - Google Patents

Description

8006414- 0 2 carboxylic acid, 78-D-mandelamido-7α-methoxy-3- [1- (Z-sulfaminoethyltetrazol-5-ythiomethyl] -B-cephem-1-carboxylic acid and 7α-methoxy-7β- (1-tetrazolylacetamido) 3- [1- (Z-sulfaminoethyltetrazol-5-ylthiomethyl] -B-cephem-1β-carboxylic acid.

The compounds of formula II are prepared by acylating 7-aminocephalosporanic acid with an appropriately protected acylating agent and then displacing the β-acetoxy group with the sulfaminoethyltetrazole thiol of formula I or a salt thereof followed by removal of the protecting group or groups, if any. The carboxylic acid group in the acylating agent is activated by any standard method, such as conversion to mixed anhydride, acid chloride, acid imidazolide or activated ester. In addition, a reagent such as dicyclohexylcarbodiimide may be used provided that the carboxylic acid group on the cephem nucleus is protected with an easily removable protecting group such as a benzhydryl, t-butyl, trichloroethyl, benzyl, benzyloxymethyl, p-methoxybenzyl nitrobenzyl ester.

When A is NH 2, the α-amino group of the acylating agent is suitably protected before the acylation with an easily removable protecting group known in the art, such as t-butoxycarbonyl, trichloroethoxycarbonyl, benzyloxycarbonyl, the methylacetoacetate adduct or similar groups commonly used in peptide synthesis. Alternatively, the compounds of formula II are prepared by acylation of a suitable 7-amino-3-sulfaminoethyltetrazolylthiomethylcephalosporin nucleus of formula III: NuzS 'S N /: 1- N 111 f-CH 2 S <|| cooR "ï- N (cnzyz -Nnso3a where W has the above definition, and Rlß with a suitable acylating agent followed by removal of any protecting groups.

The protecting groups can be removed by methods well known in the art, such as with trifluoroacetic acid, when t-butyl or t-butoxy protecting groups are used. is the hydrogen or a protecting ester group. The resulting salt is converted to the amphionic product or to the free acid by means of an ion exchange resin, such as polystyrene amine ion exchange resin (Amberlite 1R-1; or also by forming an aqueous solution of the salt. The acylating agents used as starting materials are either known or can be prepared by known methods.

The starting materials of formula III (7-amino-β-sulfaminoethyltetrazolylthiomethylcephalosporins) are prepared by reacting 7-formamidocephaloporospanic acid, prepared by reacting 7-aminocephalosporanic acid with formic acid and acetic anhydride, and sulfaminoethylene formulation with formic acid. , to remove the formyl group.

The sulfaminoethyltetrazole thiol of formula I is prepared by the reaction of 1-aminoethyl-5- (2,4-dinitrophenylthioethrazole, prepared from Zβ-dinitrofluorobenzene and 1-acetamidoethyltetrazole-5-thiol followed by acid hydrolysis of the acetamido trioxyl trimethylamine group). complex with subsequent cleavage of the 2,4-dinitrophenyl protecting group 1-Acetamidoethyltetrazole S-thiol is prepared by the reaction of an acetamidoethyl dithiocarbamate such as methyl 2-acetamidoethyldithiocarbamate and an azide such as the sodium azide. treatment of N- (Z-aminoethyllacetamide with carbon disulfide and an alkyl halide, such as methyl iodide, in the presence of a base, such as triethylamine.

Some of the compounds of formulas I, II and III may form salts with e.g. alkali metals such as sodium or potassium, alkaline earth metals such as calcium or with the ammonium cation. When A in formula II is NH 2, the compound may exist as an amphoion or as either an acid or base salt. These salts are prepared by standard procedures using a wide variety of non-toxic pharmaceutically acceptable acids and bases known in the art. Salts of the compound of formula I are considered to be the subject of the invention.

It will be appreciated that due to the asymmetric α-carbon atom of the 7-acetamido group of formula I when R 1 is O x-cH-α, α, optical isomers will exist. Racemic or split products are obtained depending on whether a racemic or split side chain acid is used as the acylating agent. The split side chain acids are readily obtained by the racemic compounds by partitioning according to well known methods, including fractional crystallization of a salt formed with an optically active acid or base.

The compounds of formula II have antibacterial activity against both gram-positive and gram-negative organisms. Minimum inhibitory concentrations (MIC) range from 0.2 to> 200 pg / ml in in vitro tests. Test results for representative compounds are given in Table 1 below. In vivo protection values for mice (5550) are given in Table 2. The compounds corresponding to the compound numbers are given in 0006414-0 4 Table 3.

Table 1 MIC (pg / ml) in vitro Compound number Bacteria 1 2 3 4 s. Aureus HH 127 3.1, 3.1 1.6 3.1 1.6 S. aureus SK 23390 0.8, 0.8 0 , 4 3.1 1.6 S. villaluz SK 70390 50, 200 25 200 200 Strep. faecalis HH 34358 100 .50 12.5 50 100 E. C011 sK 12100 0.8, 0.8 3.1 0.8 0 ,: E. coli HH 33779 3.1, 1.6 12.5 0.8 1.6 Kleb. pneumo SK 4200 1.6, 0.8 3.1 0.8 0.8 Kieb. p fi eumo s1 <12o0 0.4, 0.2 0.8 0.0 0.2 Salmonella ATCC 12176 0.2, 1.6 12.5 0.4 0.4 Shigella HH 117 0.4, 0.2 - 0.4 0.8 Pseudo. aerug. HH 63> 200,> 200> 200> 200 i> 200 Serratia, marc. ATCC 13880 100, 100> 200 200 '100 Proteus morgani 179 1.6, 3.1> 200 200 7200 Entero. aerog. ATCC 13048 50, 6.3 25 3.1 6.3 Entero. clocae HH 31254 1.6, 1.6 6.3 0.8 1.6 Table 2 ED50 (mg / kg) in vivo, E. coli SK 12140 Kleb. Pneumo SK 4200 Association number s.c. p.o. s.c. p.o. 1 0,46 50 0,46 - 2 1,02> 50 - - 3 1,56 - - - 4 1,82 50 - - 10 l5 20 25 0006414-0 s ïêbeu z Association number F föreningsnamn l 7-D-mandelamido -3- [1- (2-sulfaminoethyltetrazol-5-ylthiomethyl] -3-cephem-1 + -carboxylic acid 2 7- (2-thienylacetamido) -3- [1- (2-sulfaminoethyl) -tetrazol-5-ylthiomethyl ) -B-Cephem-11-carboxylic acid 3- 7- (1-tetrazolylacetamido) -3-1- (Z-sulfaminoethyl-tetrazol-5-ylthiomethyl) -J-cephem-1-carboxylic acid 4 7-trifluoromethylthioacetamido-3- (1- (2-Sulfaminoethyl) -tetrazol-1-ylthiomethyl) -B-cephem-carboxylic acid Pharmaceutical compositions having antibacterial activity comprise a pharmaceutical carrier containing an active but non-toxic amount of a compound of formula I. Administration may by parenteral injection, such as subcutaneously, intramuscularly or intravenously. injection of appropriately prepared sterile solutions or suspensions containing an effective, non-toxic amount of the novel cephalosporin compound is the preferred route of administration.

The compounds of formula II are prepared and administered in the same manner as other cephalosporins. The dosage comprises administering, suitably by injection, an active but non-toxic amount of a compound of formula I selected from the dosage unit range from 100 to 1,000 mg with the total daily dose ranging from # 00 mg to 6 g. The exact dosages depend on the age of the patient. and weight and the infection being treated and can be determined by those skilled in the art on the basis of the values given herein compared to the available values obtained with known cephalosporins.

The following examples illustrate the invention but are not to be construed as limiting its scope. Temperatures are in degrees Celsius unless otherwise stated. Mono-1 7-D-Mandelamido-3- [1- (2-sulfaminoethyl) tetrazo-fi-ythiomethyl) -B-cephem-11-carboxylic acid To a solution of 20.4 g (0.2O mol) of N- (Z -aminoethylacetamide in 200 ml of 9596 ethanol were added 27.9 ml (0.20 mol) of triethylamine and 12.0 ml (0.2O mol) of carbon disulfide.

The exothermic reaction reached reflux and was then cooled to ambient temperature for 1 1/2 hours. Methyl iodide (28.1 g, 0.20 mol) was added, which again produced an exothermic reaction. After 1.75 hours, the reaction mixture was evaporated to dryness and the solid residue was dissolved in 200 ml of water. The aqueous solution was extracted twice 8006M44 6 times with 250 ml portions of ethyl acetate. The extracts were combined, shaken with sodium thiosulfate, dried (MgSO 4) and evaporated to dryness to give methyl 2-acetamidoethyldithiocarbamate.

To a solution of 38.4 g (0.198 mol) of methyl 2-acetamidoethyldithiocarbamate in 100 ml of 95% ethanol was added a solution of 13.5 g (0.208 mol) of sodium azide in 100 ml of water. The reaction mixture was refluxed for 24 hours, then cooled and concentrated under reduced pressure to about half volume. The solution was cooled to ISO, and 50 ml of 6N sulfuric acid was added. The acidic solution was filtered and concentrated to about 100 ml and cooled at 5 ° to initiate crystallization of 1- (Z-acetamidoethyl) tetrazol-5-thiol, which was collected by filtration, m.p. 139-139.5 °. Additional amounts of the product were obtained by continuous extraction of the filtrate with ethyl acetate.

To a solution of 9.3 g (0.050 mol) of Zyl-dinitrofluorobenzene in 50 ml of acetone was added to a solution 9.35 g (0.050 mol) of 1- (2-acetamidoethyltetrazole-5-thiol and 6.85 ml (0.050 mol) triethylamine in 100 ml of acetone and the reaction mixture was stirred for 1 hour. The solid was collected by filtration and recrystallized from acetonitrile to give 1- (2-acetamidoethyl) -5- (2,1 + -dinitrophenylthioethrazole, mp 197-199 .

A mixture of 6.5 g (0.02 mol) of 1- (2-acetamidoethyl-5- (Zβ-dinitrophenylthio) tetrazole, 100 ml of 12N hydrochloric acid and 100 ml of 9596 ethanol was refluxed for 4.5 hours. The mixture was evaporated to dryness to give a gum-like residue which crystallized on addition of ethanol to give 1- (2-aminoethyl) -5- (2,1 + -dinitrophenylthiohetrazole hydrochloride, mp 217-219 ° (Sun.

To a mixture of 3.5 g (0.01 mol) of 1- (2-aminoethyl) -5- (2,1-dinitrophenylthio) etrazole hydrochloride in 30 ml of dry dimethylformamide was added 1.1 g (0.01 mol) ) sulfur trioxycl-trimethylamine complex followed by 1.1 ml (0.01 mol) of triethylamine.

The mixture was stirred for 1/2 hour and then filtered. The filtrate was evaporated in vacuo, acetone was added to the residue, the precipitate was removed by filtration and the filtrate was evaporated to dryness. Methanol was added to the residue and the solid formed on decomposition was removed by filtration. The methanolic filtrate was brought to pH 11.3 by adding 5% sodium methoxide in methanol, stirred for 1.25 hours, filtered and diluted with 300 ml of ether. The resulting solid was removed by filtration and the filtrate was evaporated to dryness to give a residue which was triturated with 9596 ethanol to give crystallization. The solid product was collected by filtration and dissolved in methanol, and the methanol solution was concentrated to 10 ml, diluted with 75 ml of 95% ethanol and concentrated again to 5 ml to give 1- (2-sulfaminoethyltetrazole-S-thiol disodium salt, mp 122 Calcd: 12.1966 C; 2.72% h; 216496 N 12.2596 C; 2.98% H; 217796 N En-1270.110514 25 30 eoo6414-o C3H5N5O3S2.2. solution of 1- (2-sulfaminoethyltetrazole-5-thiol disodium salt in water Found: passed through an Amberlite IR-120 ion exchange resin column to give 1- (2-sulfaminoethyltetrazole-5-thiol after lyophilization).

To a mixture of 2.71 g (0.006 mol) of 7-D-mandelamidocephalosporanic acid sodium salt and 1.18 g (0.001 mol) of 1- (2-sulfaminoethyl) tetrazole-5-thiol disodium salt in 30 ml water was added 106 g of sodium hydroxide-aqueous solution and then 596-g of sodium bicarbonate-aqueous solution to pH 7.3. The reaction mixture was heated at 700 for 2.66 hours, then cooled, covered with ethyl acetate, acidified to pH 2.5 with 3N hydrochloric acid and extracted twice with ethyl acetate.

The aqueous phase was neutralized to pH 7.0 by adding 1096 g of sodium hydroxide aqueous solution and then 596 g of sodium bicarbonate aqueous solution and chromatographed on XAD-7 resin with water and methanol as eluent.

After removal of the methanol in vacuo, the chromatography fractions were lyophilized to give 7-D-mandelamido-3- [1- (Z-sulfaminoethyl) tetrazol-5-ylthiomethyl] -3-cephem-β-carboxylic acid disodium salt C19H19N7O8S3.2 Na.2 H2 O Calculated: 3150896 C; 3.76% H; 1156496 N 3155696 C; 3.25% H; 139696 N An aqueous solution of 7-D-mandelamido-3- (1- (2-sulfaminoethyltetrazol-5): Found: thiomethyl) -3-cephem-1-carboxylic acid disodium salt was passed through a column of Amberlite IR-IZO ion exchange resin, which gave the title association.

Example 2 7- (2-Thienylacetamido) -3- [1- (Z-sulfaminoethyl) tetrazol-β-ylthiomethyl] -B-cephemolycarboxylic acid A mixture of 1.89 g (0.06l + mol) of 1- ( 2-sulfaminoethyl) tetrazole-5-thiol disodium salt and 2.67 g (0.06l + mol) of 7- (Z-thienylacetamidocephalosporanic acid sodium salt in 40 ml of water were heated at 69 ° for 5.5 hours while maintaining the pH of 7.4 by After cooling, the mixture was extracted with ethyl acetate, the aqueous phase was neutralized, evaporated to dryness and the residue was passed through an XAD-II column, eluting with water and methanol, the methanol was removed by evaporation and the water was evaporated. The solid was suspended in methanol, insoluble matter was removed by filtration and the filtrate was evaporated to dryness to give 7- (2-thienylacetamido) -3- [1- (Z-sulfaminoethyl) tetrazol-5-yltiomethyl ] -3-cephem-1β-carboxylic acid disodium salt. 80061114-0 C17H17N7O7Sw2 Na.1 CH4O Calculated: 33.90% C; 3.31% H; 15.37% N 31501196 C; 3.57% H; N 7- (2-Thienylacetamido) -3- (1- (2-sulfaminoethyl) tetrazol-5-ylthiomethyl] -3-Found: cephem-1-carboxylic acid disodium salt was converted to the title compound as described in Example 1.

Example 3 7- (1-Tetrazolylacetamido) -3- [1- (2-sulaminoethyl] tetrazol-5-ylthiomethyl) -3-cesium-1β-carboxylic acid A mixture of 3.5 g (8.5 mmol) of 7- (1- tetrazolylacetamidocephalosporanic acid sodium salt and 2.96 g (10 mmol) of 1- (2-sulfaminoethyltetrazole-5-thiol disodium salt in 50 ml of water were stirred at 650 for 6.5 hours while maintaining the pH of the reaction mixture at 7.0 by The mixture was cooled to ambient temperature, acidified to pH 1.5 with 3N hydrochloric acid, filtered and extracted three times with ethyl acetate, the pH of the aqueous phase was then adjusted to 7.0 by the addition of sodium hydrogencarbonate. , the solution was chromatographed on an XAD-Z column and the resulting product was lyophilized to give 7- (1-tetrazolylacetamido) -3- [1- (Z-sulfamino-ethyltetrazol-5-ylthiomethyl] -3-cesium-1-carboxylic acid disodium salt.

Cwl-155NUO7S32 Na.2 H2O Calculated: 263096 C; 3.37% H; 2155596 N 27.1 1% C; 3.40% H; N, 7- (1-Tetrazolylacetamido) -3- (1- (2-sulfaminoethyl) tetrazol-5-ylthiomethyl] -3-cesium-1-carboxylic acid disodium salt was converted to the title compound as Found: described in Example 1.

Example 4 7-Trifluoromethylthioacetamido-B- [1- (2-sulfaminoethyl] tetrazol-5-ylthiomethyl] -3-cephem-1-carboxylic acid A solution of 3.05 g (0.01 mol) of 1- (Z-sulfaminoethyl) tetrazole-S -thiol- disodium salt and 14.36 g (0.0l mol) of 7-trifluoromethylthioacetamidocephalosporanic acid sodium salt in 50 ml of water were heated at 700 for 5.5 hours, while maintaining the pH at 7.5 with 5% sodium bicarbonate aqueous solution. The reaction mixture was diluted with 50 ml of water and extracted twice with ethyl acetate.

The aqueous phase was acidified to pH 2 and extracted three times with ethyl acetate.

The aqueous layer was brought to pH 7.1; by adding 5% sodium bicarbonate aqueous solution and the solution was passed through an XAD-1 resin column eluting with water followed by methanol. The methanol solution was evaporated to dryness and the residue was dissolved in 75 ml of water. The aqueous solution was extracted twice with ether and petroleum ether, after which it was lyophilized. The lyophilized material was dissolved in methanol, and the solvent was evaporated to dryness and triturated with ether to give 7-trifluoromethylthioacetamido-3- [1- (2-sulfaminoethyltetrazol-5-ylthiomethyl) -3- cesium-carboxylic acid disodium salt.

C 1 H 14 F 3 N 7 O 7 Sw 2 Na 2 H 2 O Calculated: 25.149% C; 2.75% H; ll #, 8696 N Found: 25.85% C; 2.78% H; N 7-Trifluoromethylthioacetamido-B- (1- (Z-sulfaminoethyltetrazol-S-ythiomethyl) -3-cesium-1-carboxylic acid disodium salt was converted to the title compound as described in Example 1.

Example 5 7- (Da-Aminophenylacetamidw-B- (1- (2-sulfaminoethyltetrazol-5-ylthiomethyl) -3-cesium-1β-carboxylic acid - A solution of 7.58 g (0.05 mol) of 7- (Da- Butoxycarbonylaminoenyl acetamidocephalosporanic acid, 2.96 g (0.01 mol) of 1- (2-sulfaminoethyl) tetx-azole-5-thiol disodium salt and 1.26 g (0.05 mol) of sodium bicarbonate in 125 ml of water were stirred at 60 ° for 5 hours. while maintaining the pH at 7.0-7.2 by adding sodium bicarbonate, the mixture was cooled and extracted with ethyl acetate.

The aqueous phase was acidified to pH 2.5 with BN hydrochloric acid, and the acidic solution was extracted again with ethyl acetate. The aqueous phase was brought to pH 7.1 by adding 596 g of sodium carbonate solution, then passed through an XAD-11 ion exchange column and eluted with water and methanol to give 7- (Dat-butoxycarbonylaminophenylacetamido) -3- (1- ( 2-sultaminoethyl) tetrazol-5-ylthiomethyl] -B-cephem-1β-carboxylic acid disodium salt 7- (Du-L-butoxycarbonylaminoenylacetamido) -3- (1- (Z-sulfaminoethyltetetrazol-S-yltiomethyl] -3- cesium-11-carboxylic acid disodium salt is stirred at 25 ° with 25 ml of tifluoroacetic acid and 25 ml of 1J-dimethoxybenzene for 2.25 hours. The mixture is evaporated to dryness, ether is added to the residue and the precipitate is collected, washed with ether, stirred in acetonitrile for 2 hours. and dried in vacuo to give the title compound as triacetic acid salt.

An aqueous solution of the trifluoroacetic acid salt is brought to pH 5.0 by the addition of a dilute aqueous solution of sodium hydroxide. After lyophilization, the lyolized material is dissolved in methanol, and ether is added to the solution to give 7- (Da-aminophenylacetamide-B- [1- (2-sulfaminoethyltetrazol-5-ylthiomethyl) -B-cephem-1-carboxylic acid sodium salt The sodium salt is dissolved in water, and the aqueous solution is passed through an Amberlite 1R-12OH ion exchange column.

The lyophilization of the eluted material gives the title compound.

Example 6 Reaction of the Nt-butoxycarbonyl derivative of the following cephalosporanic acids: 7- (α-amino-1α-hydroxyphenylacetamidocephalosporanic acid 7- (α-amino-1β-formamidophenylacetamidocephalosporanic acid 7- (α-amino- B-formamidophenylacetamidocephalosporanic acid 7- (α-amino-11L-ureidophenylacetamidocephalosporanic acid 7- (α-amino-β-ureidophenylacetamidocephalosporanic acid 7- (α-amino-11-hydroxymethylenylacetamido) cephalosporinic acid; 7- (α-Amino-11-carboxymethylaminophenylacetamidocephalosporanic acid and 1- (Z-sulfaminoethyltetrazole-i-thiol disodium salt as described for the procedure of Example 5 followed by removal of the protecting group and conversion of the trifluoroacetic acid salts to give the following free acids: - (or-amino-1β-hydroxyphenylacetamido) -3- [1- (2-sulfaminoethyl] tetrazol-fi-ythiomethyl] -B-cephem-1β-carboxylic acid 7- (oL-amino-11-formamidophenylacetamido) -3- (1- (Z-sulfaminoethylhetrazol-5-ylthiomethyl] -Bc ephem-11-carboxylic acid 7- (α-amino-3-formamidophenylacetamido) -3- [1- (Z-sulfaminoethyl) tetrazol-fi-ythiomethyl) -3-cephem-4- [7- (oz-amino-1β- ureidophenylacetamido) -3- [1-Q-sulfaminoethyl) tetrazol-5-ylthiomethyl) -3-cephem-1β-carboxylic acid 7- (or-amino-3-ureidophenylacetamido) -3- [1- (4-Q-sulfaminoethyl) tetrazole-5- ythiomethyl) -B-cephem-11-carboxylic acid 7- (α-amino-11-hydroxymethylenylacetamido) -3- [1- (Z-sulfaminoethyl) tetrazol-S-ythiol] ethyl] -B-cephalom-11-carboxylic acid 7 - (α-amino-1,11-cyclohexadienylacetamido) -3- [10-sulfaminoethylethetrazol-5-ylthiomethyl) -B-cephem-11-carboxylic acid 7- (oL-amino-4-carboxymethylaminophenylacetamido) -3- [1 - (Z-sulfamino-ethylketrazol-S-ylthiomethyl] -B-cephem-11-carboxylic acid.

Example 7 7- (4-Hydroxymandelamido) -3- (1- (2-sulfaminoethyltetrazol-lt-ythiomethyl) -3-cephem-11-carboxylic acid is prepared by reacting 7- (4-hydroxymandel-amidokephalosporanic acid sodium salt with 1- ( 2-sulfaminoethyl) tetrazole-S-thiol disodium salt, followed by treatment of the product with Amberlite IR-IZOI-1-ion exchange resin, as described for the procedure of Example 1.

Example 8 When the sodium salt of 7- (3-sydnonacetamidocephalosporanic acid or 7- (2-aminomethylphenylacetamidocephalosporanic acid is reacted with 1- (Z-sulfaminoethyl) tetrazole-i-thiol disodium salt by the procedure described in Example 1 and the product is converted to free acid as described there, there is obtained 7- (3-sydnon-10-acetamido) -3- [1- (2-sulaminoethyl] tetrazol-5-ylthiomethyl] -3-cephem-1-carboxylic acid. acid or 7- (2-aminomethylphenylacetamido) -3- [1- (2-sulaminoethyl-tetrazol-5-ythiomethyl) -3-cephem-4-carboxylic acid. Example 9 Reaction between the sodium site of 7- (2,2, 2-Trifluoroethylthioacetamido) - cephalosporanic acid or 7-methylthioacetamidocephepiosporanoic acid and 1- (Z-sulfaminoethyltetrazole-S-thiol as described for the procedure of Example 4 gives after conversion of the formed salts to free acids 7- (2,2,2 -trifluoroethylthioacetamide-3- [1- (Z-sulfaminoethyltetrazol-5-ythiomethyl) -J-cephem-1-carboxylic acid or 7-methylthioacetamido-B- (1- (Z-sulfaminoethyltetrazol-j-ythiomethyl) yl] -3-cephem-1i-carboxylic acid.

Example 10 Reaction between a cephalosporanic acid or equivalent salt listed below: 7- (α-hydroxy-2-thienylacetamidocefioosporanic acid 7- (α-carboxy-Z-thienylacetamidocephalosporic acid 7- disodium salt by the above procedures gives, after conversion of the product to free acid, the following compounds: 7- (o-hydroxy-2-thienylacetamido) -3- (1- (2-sulfaminoethyltetrazol-5-ylthiomethyl) - 1-cephem-1-carboxylic acid 7- (α-carboxy-2-thienylacetamido) -3- (1- (2-sulfaminoethyl-tetrazo-5-ylthiomethyl) -3-cephem-11-carboxylic acid 7- (oL-sulfophenylacetamido) -3- [1- (Z-sulfaminoethyl) tetrazol-S-ylthiomethyl] -B-cephem-1-carboxylic acid.

Example 7 7-Amino-3- (1- (Z-sulfaminoethyltetrazol-1-ylthiomethyl) -B-cephem-1-carboxylic acid To a mixture of 97 g (200 ml, 2, in mol) of formic acid, distilled from anhydrous copper sulphate, and 37.5 ml (0.1 .mu.l) of acetic anhydride were added with 25.0 g (0.1 .mu.l) of 7-aminocephalosporanic acid, the mixture was stirred at ambient temperature for 0.5 hour and then evaporated to dryness. The residue was dissolved in ethyl acetate. , and the ethyl acetate solution was filtered and evaporated to dryness to give a residue which was recrystallized from ether-petroleum ether to give 7-formamidocefaiosporanoic acid.

A mixture of 1.0 g (3.3 mmol) of 7-formamidocephalosporanic acid and 0.7 g (2.6 mmol) of 1- (2-sulaminoethylketrazole-5-thio] disodium salt in 15 ml of water was stirred at 65-70 The mixture was cooled, acidified to pH 1.0 with acetic acid and extracted with ethyl acetate, the extract was filtered, and the filtrate was evaporated to dryness to give a residue, which was dissolved in methanol, the methanol solution was filtered and ether was added to precipitate the title compound, which was collected by filtration.

Example 12 7α-Methoxy-7β- (2-thienylacetamido) -3- [1- (2-sulfaminoethyltetrazol-5-ylthionethyl] -3-cephem-4-carboxylic acid A solution of 1.28 g (3 mmol) of 7α- Methoxy-7β- (2-thienylacetamido) cephalosporanic acid sodium salt is dissolved in 50 ml of water, 1.33 g (A55 mmol) of 1- (Z-suliaminoethyl) tetrazole-5-thiol disodium salt are added to the solution, and the heated to 700, until thin layer chromatography indicates that the starting material has been consumed (about 5 hours) The reaction mixture is chromatographed on XAD-L1 ion exchange resin after washing with water, with methanol as eluent, evaporation of the methanol solution gives the title compound as the disodium salt. (2-thienylacetamido) -3- [1- (2-sulaminoethyl) tetrazol-5-ylthiomethyl) -B-cetem-carboxylic acid disodium salt is converted to the title compound as described above.

Example 13 7α-Methoxy-7β-trifluoromethylthioacetamido-B- [1- (2-sultaminoethyl) tetrazol-5-ylthiomethyl] -B-cephem-11-carboxylic acid To a cold solution of 5.25 g (0, 12 mol) 7β-amino-7H-methoxycyalosporanic acid benzhydryl ester in 200 ml of methylene chloride containing 1.79 g (0.012 mol) of N, N-diethylaniline is added dropwise over 20 minutes a solution of 1.82 g (0.012 mol) trifluoromethylthioacetyl chloride in 50 ml of methylene chloride. After stirring for 30 minutes, the mixture is extracted successively with 5% aqueous sodium bicarbonate solution, 596 g of aqueous hydrochloric acid solution and finally with brine. The organic phase is dried (MgSO 4) and the solvent is evaporated to give 7OL-methoxy-7B-trifluoromethylthioacetamidocephalosporanic acid benzhydryl ester. 7α-Methoxy-7B-trifluoromethylthioacetamidocephalosporanic acid benzhydryl ester is dissolved in a cold mixture of tritluoroacetic acid-anisole (2: 1), and the mixture is stirred for 1.5 hours without external cooling. The solvent is evaporated in vacuo, and the residual product is taken up in ethyl acetate, washed with water, dried (MgSO 4) and concentrated in vacuo to a small volume. The solution is added dropwise to stirred petroleum ether to precipitate 7-methoxy-7B-trifluoromethylthioacetamidocephalosporanic acid. 7a-Methoxy-7β-trifluoromethylthioacetamidocephalosporanic acid (2.2 g, 5 mmol) is suspended in 75 ml of water, and 0.4 g of solid sodium bicarbonate is added to complete dissolution. To this solution is added 2.21 g (7.5 mmol) of 1- (Z-sulfaminoethyl) tetrazole-5-thiol disodium salt, and the mixture is heated at 700 for 7 hours.

The pH of the reaction mixture is maintained at 7.5 by dropwise addition of BN hydrochloric acid as needed. The course of the reaction is monitored by thin layer chromatography and is judged to be complete when the thin layer chromatography indicates that the starting material has disappeared. The reaction mixture is chromatographed on a column of XAD-ly resin, and the product is eluted from the column with methanol. Evaporation of the methanol solution gives 7α-methoxy-7β-trifluoromethylthioacetamido-3- [1- (Z-sulfaminoethyl] tetrazol-5-ylthiomethyl) -B-cephem-1β-carboxylic acid disodium salt.

The disodium salt is converted to the title compound by the procedures described above.

Example 14 78-D-Mandelamido-7α-methoxy-3- [1- (Z-sulfaminoethyl] tetrazol-5-ythiomethyl] -B-cephem-1s-carboxylic acid To a cold solution of 2.6 g (6 mmol) of 76-amino -7a-methoxycephalosporanic acid benzhydryl ester in 100 ml of methylene chloride containing 0.9 g (6 mmol) of N, N-diethylaniline dichloroacetylmandeloyl chloride in 25 ml of methylene chloride The reaction mixture is allowed to add dropwise over 15 minutes a solution of 1.7 g (6 mmol) DO- assume room temperature with stirring, after which it is extracted successively with 596 μg of aqueous sodium bicarbonate solution, 596 μg of hydrochloric acid and brine.

The organic phase is dried and evaporated in vacuo. The residue is dissolved in cold trifluoroacetic acid anisole (2: 1), and the mixture is stirred at ambient temperature for 1 hour. The mixture is evaporated in vacuo, and the residue is dissolved in 596% aqueous sodium bicarbonate solution. The pH is raised to 9-9.3 by adding 596 g of aqueous sodium carbonate solution and kept there for 30 minutes so that the cleavage of the dichloroacetyl group is complete. The solution is cooled in ice, layered with ethyl acetate and acidified to pH 2.0 with dilute hydrochloric acid. The layers are separated, and after a second extraction of the aqueous layer with ethyl acetate, the organic phases are combined, dried and evaporated in vacuo to give 7B-D-mandelamido-7a-methoxycetalosporanic acid. 7β-D-MandelamidoJa-methoxycephalosporanic acid (2.2 g, 5 mmol) is suspended in 75 ml of water, and sodium bicarbonate is added until all the acid has dissolved.

To this is added 2.21 g (7.5 mmol) of 1- (Z-sulfaminoethyl) tetrazole-thiol disodium salt and the mixture is heated at 700 for 7 hours. The pH of the reaction mixture is maintained at 7.5 by the addition of 3N hydrochloric acid. solution of XAD-lf resin during dilution with methanol gives, after evaporation of the methanol, the title compound as its disodium salt 7B-D-Mandelamido-Ja-methoxy-B- fl- (Z-sulfaminoethyltetrazol-S-yltiomethyl] -B-cephem-1β-carboxylic acid disodium salt is converted to the title compound by the procedures described above: GOOGMIi-Û lit Example 15 7u-Methoxy-7B- (D-α-aminophenylacetamido) -3- [1- ( Z-sulfaminoethylethetrazol-S-ylthiomethyl) -B-cephem-11-carboxylic acid To a solution of 5.3 g (0.0122 mol) of 7β-amino-7α-methoxycephalosporanic acid p-nitrobenzyl ester in 200 ml of methylene chloride is added 3.0 g (0.02 mol) Da-L-butoxycarbonylaminophenylacetic acid and 2.5 g (0,02 mol) dicyclohexylcarbodiimide The mixture is stirred for 18 hours at ambient temperature and then filtered. t is evaporated in vacuo, and the residue is dissolved in methanol-tetrahydrofuran and hydrogenated over 5% palladium on carbon to give 7B- (D-α-t-butoxycarbonylaminophenylacetamido) -7a-methoxycephalosporanic acid. 76- (D-α-t-butoxycarbonylaminophenylacetamido) -7α-methoxy-cephalosporanic acid (2.68 g, 5 mmol) is dissolved in 75 ml of water by adding 0.4 g of solid sodium bicarbonate. 1- (2-Sulfaminoethyl) tetrazole-5-thiol disodium salt (2.2l g, 7.5 mmol) is added, and the reaction mixture is heated at 700 until thin layer chromatography indicates that the starting material has disappeared. The reaction mixture is chromatographed on XAD-II resin and eluted with methanol. Evaporation of the methanol solution gives 7α-methoxy-7β- (D-oL-t-butoxycarbonylaminophenylacetamido) -3- [1- (Z-sulfaminoethyltetrazol-5-ylthiomethyl] -B-cephem-1-carboxylic acid disodium salt.

The disodium salt is suspended in 1: 1 trifluoroacetic acid anisole and stirred at ambient temperature for 2 hours. Excess trifluoroacetic acid is removed by evaporation. The residue is triturated with ether, and the resulting precipitate is collected by filtration and stirred with acetonitrile to give the title compound as its trifluoroacetic acid salt.

An aqueous solution of the trifluoroacetic acid salt is brought to pH 7 by adding 596 g of aqueous solution of sodium bicarbonate and then chromatographed on XAD-II resin with methanol as eluent. The solid obtained after evaporation of the methanol is dissolved in water, and the aqueous solution is passed through a cation exchange column (IR-IZOH). Lyophilization of the eluted material gives the title compound.

Example 16 7u-Methoxy-7B- (1-tetrazolylacetamide-B- [1- (2-sulfamidoethyltetrazol-5-ylthiomethyl) -B-cephem-1β-carboxylic acid Yield to an equivalent amount of 1-tetrazolylacetyl chloride Example 13 gives 7α-methoxy-7β- (1-tetrazolylacetamido) -cephalosporanic acid benzhydryl ester, which is converted to 7α-methoxy-7β- (1-tetrazolylacetamidolcephalosporanic acid as described therein.

Reaction between 7u-methoxy-7β- (1-tetrazolylacetamidohcephalosporanic acid, 1- (2-sulfaminoethyl) tetrazole-5-thiol disodium salt and sodium bicarbonate such as

Claims (1)

1. w06414-0 15 described in Example 13 gives, after conversion of the disodium salt product to free acid as described above, the title compound. PATENE REQUIRED Intermediate for use in the preparation of therapeutically valuable compounds of the formula u 1 I ä's of-N / C323 / coon NN (cnzlz -nnso3a where W is hydrogen or methoxy, RI is: S? 9 l? X Cn-c-, Y-cHZ-c- or z- fl mm-cne-g. A A is 10 thienyl, dihydrophenyl, phenyl or phenyl monosubstituted with hydroxy, hydroxymethyl, formamido, ureido or carboxymethylamino, A is NHZ, OH, COOH, or SO 3 H, or formyloxy when X is phenyl, Y is thienyl, tetrazolyl, sydnone, cyano or aminomethylphenyl, Z is methyl, trifluoromethyl, trifluoroethyl, cyanomethyl or pyrldyl, and 15 m is zero to two, or a toxic pharmaceutically acceptable salt thereof, characterized in the formula un ns- If-_N (C325 -Nnsoau