HU179967B - Process for preparing 7-acyl-amido-3-thiomethyl-3-cefem-carboxylic acid derivatives - Google Patents

Abstract

The invention relates to a process for the preparation of 3- (thiomethyl) cephalosporins. For example, 3- ((W-ethyl-1-1H-tetrazol-5-yl) thio) methyl) -7- (2- (2-thienyl) acetamide) -3-c ephem-A-carboxylic acid is used in the present invention in acetonitrile, by heating трт 7- (2 "· (2-thienyl) acetamido) cephalosporanic acid and 1-methyl-1-n-tetrazole-5-thiol in dry acetonitrile at reflux temperature, then evaporating the reaction mixture to remove the solvent and the ethanol and dicyclohexylamine are added to the resulting residue, whereby the desired product precipitates as Dicyclohexylaminsalζ.

Description

Process for the preparation of 7-acylamido-3-thiomethyl-3-cephem-4-carboxylic acid derivatives

This invention relates to the displacement of an acetoxy group of 7-acylamido-3-acetoxymethyl-3-cephem-4-carboxylic acids (cephalosporanic acids) with a sulfur-containing nucleophilic reagent using an organic solvent and anhydrous conditions.

Substitution of the acetoxy group of 7-Acyl-α-amido-3-acetoxymethyl-3-cephem-4-carboxylic acids with a sulfur-containing nucleophile reagent is known from U.S. Patent No. 3,278,531. According to this patent and other publications (J. D. Cocker, J. Cbem. Soc. 1965, 5015), the reaction takes place only in aqueous medium. The substitution reactions were performed in water and the salt of the starting cephalosporanic acid and the sulfur-containing nucleophilic reagent or a salt thereof were reacted in an aqueous medium at pH 5-8. In many cases, the combination of the aqueous medium, the higher temperature (35-70 ° C) and the near neutral or alkaline pH caused the decomposition of the cephem core, and the products obtained in this way often required a high degree of purification. Attempts have been made to displace the cephalosporanic acid in water at low pH (pH = 2-3), but as a side reaction there is a high level of lactone formation which impairs the yield of the desired product.

It has been found that the acetoxy group of cephalosporanic acids (and other 3-acyloxy groups) can be substituted by the sulfur-containing nucleophilic reagent in an organic solvent under anhydrous conditions. Reactions carried out in this way are not accompanied by lactone formation, the yield is generally higher and the products are more easily isolated. Some products will spontaneously precipitate out of the reaction mixture. It is contemplated that the process of the invention is generally applicable and applicable to any sulfur-containing nucleophilic reagent and cephalosporanic acid.

The present invention relates to an improved process for the preparation of 7-acylamido-3-thiomethyl-3-cephem-4-carboxylic acids (i.e., (3-thiomethyl) -cephalosporins).

The present invention relates to a novel process for the preparation of compounds of formula I wherein:

R ^{13 is} amidino, benzoyl, phenyl, C 1 -C 4 alkyl, or 4;

R ^{1 is} hydrogen or methoxy;

R ² is a group of formula 6; wherein R ³ is -CH ₂ - (1-3 chloroalkyl) -alkyl,

1-4 Cyano-cyanoalkyl, 4-amino-4-carboxy-20-butyl optionally protected by a halo-substituted benzoyl group, wherein

¹ represents an amino group protected as above - a group of the formula 8 wherein a and ¹ independently represent hydrogen or hydroxy and m = 0 or 1; tetrazolyl-methyl; A group of formula 9 wherein

P is thienyl or phenyl of formula 10 wherein a and ¹ are as defined above and and

Q is a hydroxy or formyloxy group, an amino group protected by a tert-butoxy30 carbonyl group, or

-1179967

Represents an acylated amino group of the formula: wherein

T 13 is a group;

¹³ report, the group R 4 of the formula R ⁶ is a unit with which the group of formula 22 5-optionally substituted 5- or 6-membered one nitrogen and two oxygen or sulfur, or three or four nitrogen atoms in a heteroaromatic ring, or means a unit by which the group of formula 22 forms a 2-benzothiazolyl group. 10

The process of the invention is carried out by reacting a compound of formula II wherein R ¹ and R ² are as defined above and

R 1 is C 1-3 alkyl, C 4-6 cycloalkyl, amino, mono- or di- (C 1-3) alkyl-15-amino, 24, 25 or 26; With a sulfur-containing nucleophilic reagent of formula R ¹³ -S-R ¹⁴ - wherein

R ¹³ is as defined above and R ^{14 is} hydrogen or 20 when R ^{13 is} methylene-ammonium,

R ¹⁴ in combination with R ¹³ forms thiourea - in an organic solvent, under anhydrous conditions.

The compounds of the present invention exhibit antibacterial activity; moreover, most of them can be used as intermediates for the preparation of other cephalosporins with antimicrobial activity (see, for example, U.S. Patent No. 3,932,393).

Both the 3-acyloxymethyl-cephalosporins of formula II and the amino groups of the sulfur-containing nucleophilic reagent are preferably protected. Methods of protecting amino groups are well known (Protective Groups in 35 Organic Chemistry, ed. JTW McOmie, Plenum Press, London and New York, 1973). Generally, amino group protection involves the removal of protecting groups; however, this is not necessary for the 7-acylamido group of formula 27, which can subsequently be cleaved from the cephalosporin. Suitable amino protecting groups include, for example, C ₂ -C ₄ alkanoyl, C ₃ -C ₄ chloro or fluoroalkanoyl, benzoyl and substituted benzoyl. "C ₂ -C ₄ alkanoyl" may be acetyl, propyl, butyryl and the like. The term "C ₃ -C ₄ chloro or 45 fluoroalkanoyl" refers, for example, to 3-chloropropionyl, 3-fluoropropionyl. By "substituted benzoyl" is meant, for example, 4-chlorobenzoyl, 4-bromobenzoyl and 2,4-dichlorobenzoyl. These groups may also be used to protect amino groups present in sulfur-containing nucleophilic reagent 50 and other positions of 3-acyloxymethyl-cephalosporins. But if it is necessary to liberate the amino groups, an easily removable protecting group should be used (see McOmie, cited above). Easily removable protecting groups include benzyloxycarbonyl, p-nitrobenzyloxycarbonyl, 2,2,2-trichloroethoxycarbonyl, tert-butoxycarbonyl, ρ-methoxybenzoyl. oxycarbonyl; and diphenylmethoxycarbonyl. The amino group of formula 27 can also be protected according to Belgian Patent No. 771,694, 60.

By "halogen" is meant fluorine, chlorine, bromine or iodine. The term "C 1-4 alkoxy" refers, for example, to methoxy, ethoxy, isopropoxy, n-butoxy 65. The term "alkali metal" preferably includes sodium, potassium and lithium.

Examples of the groups of formula 6 wherein R ^{3 has} the meaning given above are, for example, formamido, acetamido, propionamido, butyramido, alpha-methylpropionamido, valeramido, alpha-methylbutyramido, trimethylacetamido, hexanamido, heptanamido, 3-chloropropionamido, 3-chlorobutyramido, 4-fluorobutyramido, 5-chloro valeramido, cyanoacetamido, 2-cyanopropionamido, , 3-cyanopropionamido, 4-cyanobutyramido, hydroxyacetamido, 2-hydroxypropionamido, 3-hydroxybutyramido, ρ-nitrobenzyloxycarbonylamino, tert-butoxy, -carbonylamino, (2,2,2-trichloroethoxy) carbonylamino, 5- (2,5-dichloro-benzamido) -5-carboxy-valeramido, 5-acetamido-5-carboxy-; valeramido, 5-carbomethoxy-5- (2,4-dichloro-benzamido) -valeramido, 5-carbon-n-butoxy-5- (2,4-dichloro-benzamido) -valeramido, 5-carboxy-5 - (2,4-dichlorobenzamido) -valeramido, 5- (p-chlorobenzamido) -5-carboxy-valeramido, 5-propionamido-5-carboxy-valeramido, 5- (3-chloropropionamide) ) -5 -carboxy-valeramido, 5-benzamido-5-carboxy-valeramido, 5-oxo-5-carboxy-valeramido and 4-carboxy-butyramido.

For groups of formula 6, wherein

R ³ is a group of the formula wherein the latter groups include, by way of example, phenylacetamido, 2- (p-hydroxyphenyl) acetamido, 2- (m-hydroxyphenyl) acetamido- and 2- (m-hydroxyphenyl) acetamido.

When m = 1 in the above formula, a typical group is phenoxyacetamido.

When R ³ 9 is a group, typical examples of 6 are:

2-OH-2-phenylacetamido and its O-formyl derivative; protected 2-amino-2-phenylacetamido - where the protected amino group is, for example, carbonylamino, tert-butoxycarbonylamino, trichloroethoxycarbonylamino or p-nitrobenzyloxycarbonylamino -; and 2- (acylated amino) -2-phenylacetamido. These groups include the 2-substituted 2-thienylacetamido and 3-thienylacetamido groups where the phenyl group is replaced by a 2-thienyl or 3-thienyl ring.

Typical members of the above acetamido groups are 2-OH-2-phenylacetamido, 2-p-hydroxyphenyl-2-hydroxyacetamido, 2-m-hydroxyphenyl-2-hydroxyacetamido, 2-O-formyl-2. -phenylacetamido, 2-hydroxy-2- (3-thienyl) acetamido, 2-formyloxy-2- (2-thienyl) acetamide, 2-formyloxy-2- (3- thienyl) -acetamido, 2- (p-nitrobenzyloxycarbonylamino) -2-phenylacetamido, 2- (tert-butoxycarbonylamino) -2- (2-thienyl) acetamide -, 2- (benzyloxycarbonylamino) -2- (m-hydroxyphenyl) acetamido-, 2-tert-butoxycarbonylamino) -2- (p-hydroxyphenyl) acetamido- 2-ureido-2-phenylacetamido, 2-ureido-2- (2-thienyl) acetamide and 2- (3-guanyl-1-ureido) -2-phenylacetamido.

Most of the cephalosporanic acids used as starting materials for the process of the invention are known compounds and can all be prepared using methods well known in the art (Cephalosporins and Penicillins, Chemistry and Biology, ed. Flynn, EH, Academic Press, New York, 1972). , Chapters 4 and 15 and the literature cited therein See also U.S. Patent Nos. 3,914,157 and South African Patent Application 71/3229).

TTI is a sulfur containing nucleophilic reagent

Among the residues -2179967, those wherein R ¹³ is 4 are preferred.

Of the groups of formula 4, we prefer the following 5-membered heteroaromatic rings:

1.2.3-Oxadiazole

1.2.4-Oxadiazole

1.2.5-Oxadiazole

1,3,4-oxadiazole

1.2.3- Thiadiazole

1.2.4- Thiadiazole

1.2.5- Thiadiazole

1,3,4-thiadiazole

1.2.3- Triazole

1.2.4- Triazole tetrazole

Suitable 6-membered rings include pyrazine

1.2.3- Triazine

1.2.4-triazine

1.3.5-triazine

1.2.3.4-Tetrazine

1.2.4.5- tetrazine

Certain heteroaryl thiols of formula III, as well as certain alkyl thiols and phenyl thiols, exist in the form of both thions and tautomeric mixtures of thiols and thions. For example, 2-methyl-1,3,4-thiadiazol-5-yl-thiol exists in tautomeric forms a and b. However, the reaction of the present invention with the said group of sulfur-containing nucleophilic reagents will in any case take place, either in thiol or thionic form. Accordingly, it is within the scope of the present invention to carry out the reaction with a reagent in the form of thion or thiol / thionomer.

The heteroarylthiol may be unsubstituted or may contain one or more substituents. In general, the quality of the substituents is not critical. Best results are obtained when all primary or secondary amino groups are protected. If the heteroaryl thiol contains more than one thiol group, the thiol group which is not desired to be reacted must be protected. There are known methods of protecting thiols (see, for example, McOmie, cited above). When protecting these groups, the reaction proceeds regardless of the nature of the substituents.

It is preferable to avoid too many and large substituents; it is generally preferred that the substituents have a molecular weight of not more than 500. Typically, the substituents of conventionally substituted heteroaromatic rings have a total molecular weight of less than 250.

Suitable substituents include those wherein the substituent is C 1-4 alkyl, benzyl, phenyl, oxo, hydroxyl, benzyloxycarbonylaminomethyl, carboxymethyl or N-methylacetamido.

Typical sulfur-containing nucleophilic reagents of formula III include, for example:

Thioureas Allyl thioureas

N, N'-di-n-butylthiourea N, N'-di-tert-butylthiourea N, N'-dicyclohexylthiourea N, N'-diisopropylthiourea N, N '-dimethyl- thiourea N-methylthiourea

Ν, Ν, Ν ', Ν'tetraethyl thiourea

Ν, Ν, Ν ', N'-tetramethylthiourea thiourea

N-phenyl-thiourea

N, N '-diphenylthiourea

N, N-dimethyl-thiourea

thiobenzoic

Alkylthiols Methanethiol Ethanethiol

1- or 2-propanethiol

1- or 2-butanethiol

2-methyl-p-propanethiol

thiophenol

Heteroaryl thiols

1-methyl-1,2,3-triazol-5-yl-1-thiol

2-methyl-1,2,3-triazol-4-yl-thiol

3,5-dimethyl-l, 2,3-triazol-4-yl-thiol

1.2.4-Triazol-3-yl-thiol

4-methyl-1,2,4-triazol-5-yl-thiol

3-methyl-1,2,4-triazol-5-yl-thiol

3,4-Dimethyl-1,2,4-triazol-5-yl-thiol

2-methyl-1,2,4-triazol-5-yl-thiol

2-benzyl-1,2,4-triazol-5-yl-thiol

2,3-dimethyl-1,2,4-triazol-5-yl-thiol

3- (Carboxymethyl) -4-methyl-1,2,4-triazol-5-yl-thiol

3-hydroxy-4-methyl-l, 2,4-triazol-5-yl-thiol

5-methyl-1,2,3-oxadiazol-4-yl-thiol

1.2.4-Oxadiazol-3-yl-thiol

3-methyl-1,2,4-oxadiazol-5-yl-thiol 3-methyl-1,2,5-oxadiazol-4-yl-thiol

2-methyl-1,3,4-oxadiazol-5-yl-thiol

3-methyl-1,2,4-thiadiazol-5-yl-thiol

1.3.4-Thiadiazol-5-yl-thiol 2-Methyl-1,3,4-thiadiazol-5-yl-thiol

2- (N-Methylacetamido) -1,3,4-thiadiazol-5-ylthiol 5-Tetrazolylthiol

-methyl-tetrazol-5-yl-thiol 1-benzyl-tetrazol-5-yl-thiol 1- (carboxymethyl) -tetrazol-5-yl-thiol

1- (2-Dimethylaminoethyl) -tetrazol-5-yl-thiol

1.2.3-Triazin-4-yl-thiol

1.2.4-Triazin-3-yl-thiol

4.5-Dihydro-6-hydroxy-4-methyl-5-oxo-1,2,4-triazin-3-yl-thiol

1.3.5-Triazin-2-yl-thiol

1.2.4.5-Tetrazin-3-yl-thiol

2-benzimidazolylthiol

2-benzofuran-tiazoliltiol

2-Benzoxazolylthiol tetrazolo [1,5-b] pyridazin-6-yl thiol

Most of the sulfur-containing nucleophilic reagents used in the process of the invention are known compounds, and each can be prepared by known methods. For the preparation of heteroaryl thiols, reference is made to several volumes of Heterocyclic Compounds, ed. Robert C. Elderfield, John Wiley and Sons, Inc., N.Y., and The Chemistry of Heterocyclic Compounds, ed. Weissberger et al., John Wiley and Sons, N.Y.

It is critical for the process of the invention to carry out the reaction in an organic solvent. However, the type of solvent is not critical as it is numerous

-3179967 organic solvent was found to be satisfactory. In general, the following classes of solvents are preferred: aliphatic and aromatic hydrocarbons, alcohols, amides, ethers, ketones, carboxylic acids, carboxylic acid esters, halogenated hydrocarbons, nitro compounds, nitriles and thioethers. Because some nucleophilic reagents are liquid, they can also be used as solvents when used in excess.

The solvent must be inert in the sense that it will not react with any of the reagents. Preferred solvents are: Hydrocarbons:

benzene toluene ο-, m- or p-xylene cumene mesitylene

Halogenated hydrocarbons:

chloroethane

1- or 2-chloropropane

1- or 2-chlorobutane isobutyl chloride

1,2- or 3-chloropentane methylene chloride chloroform

1.2-dichloroethane carbon tetrachloride

1.1.1-trichloroethane

1.1.2-tri chloroethane

1.1.2.2- tetra chloroethane fluorobenzene chlorobenzene bromobenzene ο-, m- or p-chlorotoluene ο-, m- or p-bromotoluene ο-, m- or p-dichlorobenzene

acids:

acetic acid propionic acid butyric acid isobutyric acid valeric acid

Esters:

methyl acetate ethylene glycol diacetate ethyl acetate propyl acetate isopropyl acetate butyl acetate isobutyl acetate sec-butyl acetate amyl acetate ethyl propionate methyl butyrate n-butyl formate propylene carbonate ethylene carbonate

Nitro Compounds:

nitrobenzene nitromethane nitroethane nitropropane

nitrites:

acetonitrile propionitrile butyronitrile isobutyronitrile valeronitrile benzonitrile t'enylacetonitrile

ketones:

methyl isobutyl ketone methyl ethyl ketone

Particularly preferred solvents are acetonitrile, 1,2-dichloroethane, methylene chloride, propionhitrile, nitromethane, nitroethane, acetic acid, isopropyl acetate, butyl acetate and methyl isobutyl ketone.

The 3-acyloxymethyl-cephalosporin of formula II is defined as an acid. It is believed that the acid reacts because cephalosporin salts are generally insoluble in organic solvents. An exception is the solubility of cephalosporin salts in carboxylic acids such as acetic acid. Therefore, the reaction according to the invention is carried out by dissolving 3-acyloxymethyl-cephalosporin salts, such as metal salts, in a carboxylic acid solvent.

Regardless of the type of solvent, the reaction according to the invention must be carried out under anhydrous conditions. Generally, the reaction mixture contains less than 5% water, preferably less than 1% water. More preferably, the water content is less than 0.5%. If commercial grade reagents and solvents are not sufficiently anhydrous, water is removed by known methods, including azeotropic distillation and drying agents such as alumina, silica gel, anhydrous calcium sulfate, and the like.

The reaction according to the invention can be carried out over a wide range of temperatures. In general, a reaction temperature of 50-140 ° C may be used, but better results are obtained at 70-120 ° C. The reaction can be carried out at higher pressures, but the advantage has not been found. Therefore, atmospheric pressure is preferred for its simplicity.

The amount of reagents is not critical. It is generally preferred to use an excess of sulfur-containing nucleophilic reagent, for example, 1.0 to 5.0 molar equivalents of nucleophilic reagent for 1 molar equivalent of 3-acyloxymethyl-cephalosporin.

The process of the invention is particularly suitable for introducing into the cephalosporins the following 3-heteroarylthio groups: 5-methyl-1,3,4-thiadiazol-2-yl-thiol-methyl-1H-tetrazol-5-yl-thio-1-yl. carboxymethyl-1H-tetrazol-5-ylthio-

4-methyl-6-hydroxy-5-oxo-l, 2,4-triazin-3-ylthio

1-methyl-1,3,4-triazol-2-yl-thio-

2-methyl-1,3,4-triazol-5-yl-thio-

1,2-dimethyl-1,3,4-triazol-5-yl-thio-

3-methyl-1,3,4-triazol-5-yl-thio-

2,3-Dimethyl-1,3,4-triazol-5-yl-thiol-methyl-2-trifluoromethyl-1,3,4-triazol-5-yl-thiol-methyl-2-carboxamido-1, 3,4-Triazol-5-yl-thiol-methyl-2-carboxymethyl-1,3,4-triazol-5-yl-thio-4179967-1-methyl-2-carboalkoxy-1,3,4 -triazol-5-yl-thiol-methyl-2- (protected aminomethyl) -1,3,4-triazol-5-yl-thiol-methyl-2- (protected amino) -1,3,4-triazole -5-yl-thiol-methyl-2-hydroxy-1,3,4-triazol-5-yl-thio-1H-tetrazol-5-yl-thio-1- (2-dimethylamino-ethyl) -1H-tetrazole -5-ylthio

1- (sulfomethyl) -1H-tetrazol-5-ylthio, alkali metal salt

5-methyl-J, 4-oxadiazol-2-ylthio

1,3,4-tiúdiazöl-2-ylthio

3-methyl-1,2,4-thiadiazol-5-ylthio;

2- (protected aminomethyl) -1,3,4-thiadiazol-5-ylthio

Important known cephalosporin compounds for the synthesis of the present invention include, for example:

cefamandol: compound c: cefarttandol naphthalate: compound d: cefazolin: compound e: ceftezole: compound cefazaflur: g compound SCE-963: h compound BLS-786: i compound CS-1170: j compound SQ- 67,590: k Compound SKF-75,073: BBS-226: m SQ-14,359 Compound

The products of the present invention, their unprotected derivatives and their pharmaceutically acceptable salts can be used to combat infections in warm-blooded mammals in parenteral, non-toxic doses of about 10 to 500 mg / kg body weight. The compounds are processed into pharmaceutical compositions in the conventional manner.

The following examples illustrate the preparation of the compounds of the invention.

Example 1

Preparation of 3 - {[(1-Methyl-1H-tetrazol-5-yl) thio] methyl} -7- [2- (2-thienyl) acetamido] -3-cephem-4-carboxylic acid in acetonitrile

7- [2- (2-Thienyl) acetamido] cephalosporanic acid (2.2 g;

5.5 mmol) and 1-methyl-1H-tetrazole-5-thiol (1.0 g;

8.6 mmol) to 25 ml of acetonitrile in a flask equipped with a condenser with anhydrous calcium sulfate ("Drierite"). The reaction mixture is heated to reflux with stirring and the progress of the reaction is monitored by thin layer chromatography (TLC). After refluxing for 90 minutes, TLC showed about ^2/3 parts of the cephalosporin starting material and V3 parts of the desired product. After 3 hours at reflux, TLC indicates about V3 parts cephalosporin starting material and ^2/3 parts product. After 6 hours, when TLC indicates complete reaction, the reaction mixture was cooled to room temperature and allowed to stand overnight. The solvent was removed by rotary evaporation and the residual foam was dissolved in 10 mL of ethanol. After the dropwise addition of 1 ml of dicyclohexylamine in 10 ml of ethanol, the product precipitates as the dicyclohexylamine salt which is filtered off after 15 minutes of stirring. The isolated product was washed with 15 ml of ethanol and dried in vacuo at 45 ° C for 2 hours to give 2.50 g (76.1% yield);

mp 183-184 ° C (dec.). The product was analyzed by nuclear magnetic resonance (NMR), infrared (IR) spectroscopy, and layer chromatography (TLC), and the results were identical to the analysis of a credible product prepared by a known aqueous substitution reaction.

NMR (DMSO-d6):

δ 3.52 (m, 2, 2-C ₂ ),

3.76 (s, 2, -C / 7 ₂ CONH-),

3.92 (s, 3, tetrazole-C ₃ ),

4.35 (s, 2, 3-C / 7 ₂ S).

5.00 (d, 1, C ₆ -ti, J = 5 Hz),

5.55 (q, 1, C ₇ -ti, j = 5 Hz, J = 9 Hz),

6.95 (d, 2, thiophene 3- and 4- / 7, j = 3 Hz),

7.35 (t, 1, thiophene 5-H, J = 3 Hz),

8.75 (d, 1, -CH ₂ C ₆ N 7 -, J = 9 Hz).

Example 2

Preparation of 3 - {[(1-Methyl-1H-tetrazol-5-yl) thio] methyl} -7- [2- (2-thienyl) acetamido] -3-cephem-4-carboxylic acid in acetonitrile

7- [2- (2-Thienylacetamido] cephalosporanic acid (4.0 g; 10 mmol) and 1-methyl-1H-tetrazol-5-thiol (5.8 g; 50 mmol) in 50 mL of anhydrous acetonitrile (" Amberlite 15 "anhydrous sulfonic acid resin treated), a dry atmosphere boiled for 8 ^3/4 hours at reflux temperature. the reaction was followed by TLC and found that at the end of 8 ^3/4 hours, the reaction was essentially complete.

The solvent was evaporated and ethanol (125 mL) was added to the residue. After addition of 6 ml of dicyclohexylamine in 75 ml of ethanol, the product precipitates as the dicyclohexylamine salt. Filtration separated, washed and dried gave 4.50 g (71% yield) of product. IR, NMR, and TLC showed the product to be identical to the authentic sample prepared by aqueous substitution. The NMR spectrum was identical to that of Example 1.

Example 3

Preparation of 3 - {[(1-Methyl-1H-tetrazol-5-yl) thio] methyl} -7- [2- (2-thienyl) acetamido] -3-cephem-4-carboxylic acid 1,2-dichloro ethane

7- [2- (2-Thienyl) acetamido] cephalosporanic acid (2.0 g, 5 mmol) and 1-methyl-1H-tetrazol-5-thiol (1.2 g, 10 mmol) in 25 mL of 2 in dichloroethane under reflux for a total of 5.5 hours. The solvent was then removed on a rotary evaporator and 25 mL of ethanol was added to the residue followed by dropwise addition of 2 mL of dicyclohexylamine in 25 mL of ethanol. The product crystallizes in the form of its dicyclohexylamine salt, which is stirred for 20 minutes at room temperature, then filtered, washed with 25 ml of ethanol and dried at 40 ° C in vacuo. 2.34 g of an off-white solid are obtained (73.8% yield). IR and NMR spectra; zones with authentic samples obtained by substitution in aqueous medium. A. The NMR spectrum is identical to that of Example 1.

-5179967

Example 4

Preparation of 3 - {[(1-Methyl-1H-tetrazol-5-yl) thio] methyl} -7-amino-3-cephem-4-carboxylic acid

1,2-dichloroethane

7-Formamidocephalosporanic acid (3.0 g; 10 mmol) and 1-methyl-1H-tetrazole-5-thiol (2.4 g; 20 mmol) in 50 mL

It is stirred in 1,2-dichloroethane and refluxed for 7 hours. The reaction mixture was then cooled to room temperature and allowed to stand overnight at room temperature. A solid red gummy material precipitates. The product is identified by removing the 7-formyl group as follows:

The solvent was removed on a rotary evaporator and the residue was dissolved in methanol (25 mL) and concentrated hydrochloric acid (2.8 mL) and allowed to stand at room temperature overnight. The solution was then diluted to 50 ml with water and the initial pH of 0.9 was increased to 3.6 by the addition of triethylamine. The resulting light brown crystals were collected by filtration, washed with water and dried to give 2.10 g (64% yield) of product. The NMR spectrum was identical to that obtained by substitution in an aqueous medium.

NMR (D ₂ O, NaHCO 3):

δ 3.65 (q, 2, 2-C / 7 ₂ , J _ab = 16.5 Hz),

4.08 (s, 3, -CH ₃ in tetrazolone),

4.16 (q, 2, 3-Cff ₂ S, J _AB = 12.5 Hz),

5.05 (d, 1, C ₆ -H, J = 5 Hz) and

5.45 (d, 1, C ₇ -H, J = 5 Hz).

Example 5

Preparation of 3 - {[(1-Methyl-1H-tetrazol-5-yl) thio] methyl} -7- [2- (2-thienyl) acetamido] -3-cephem-4-carboxylic acid in isopropyl acetate

7- [2- (2-Thienylacetamido) -ephephalosporanic acid (2.0 g;

and 1-methyl-1H-tetrazole-5-thiol (1.2 g, 10 mmol) in isopropyl acetate (25 mL) was refluxed (90 ° C) for 23 hours.

The reaction mixture was then cooled to room temperature, and little cephalosporanic acid starting material remained by TLC. The light cream solid was collected by filtration, washed with isopropyl acetate and dried to give 1.60 g (70.8% yield) of product. NMR confirmed product identity and found that cephalosporanic acid starting material is present in an amount of less than 1%.

NMR (DMSO-dg):

δ 3.72 (s, 2, 2-CH ₂ ),

3.80 (s, 2, -Ctt ₂ CO ₂ NH),

3.95 (s, 3, -CH ₃ tetrazole),

4.30 (s, 2, 3-CTF ₂ S-)

5.10 (d, 1, C ₆ -H, J = 5 Hz),

5.70 (q, 1, C ₇ -H, J = 5 Hz, J = 8 Hz),

6.92 (d, 2, thiophene 3- and 4-H, J = 3 Hz),

7.35 (t, 1, thiophene 5-H, J = 3 Hz) and

8.78 (d, 1, -CH ₂ -CON / 7-, J = 8 Hz).

Example 6

Preparation of 3 - {[(1-Methyl-1H-tetrazol-5-yl) thio] methyl} -7- [2- (2-thienyl) acetamido] -3-cephem-4-carboxylic acid in propionitrile

7- [2- (2-Thienyl) acetamido] cephalosporanic acid (2.0 g;

millimoles) and 1-methyl-1H-tetrazole-5-thiol (1.2 g;

millimoles) in 25 ml of anhydrous propionitrile was refluxed (97 °) until TLC indicated complete disappearance of the cephalosporanic acid starting material (4.5 hours). The reaction mixture was then cooled to room temperature and the solvent removed on a rotary evaporator; during this operation, 15 small portions of the solution are lost due to foaming. The residue was dissolved in warm ethanol (35 ml) and dicyclohexylamine (2 ml) dissolved in ethanol (10 ml). The product precipitated as the dicyclohexylamine salt was stirred for 10 minutes at room temperature and filtered, washed with ethanol and dried, yielding 1.24 g (39.0%, after deduction of loss on rotary evaporator). NMR analysis showed the product to be identical to the product of Example 1.

Example 7

Preparation of 3 - {[(1-Methyl-1H-tetrazol-5-yl) thio] methyl} -7- [2- (2-thienyl) acetamido] -3-cephem-4-carboxylic acid in acetonitrile, 3 With the addition of 5-dichlorophenyl dihydrogen phosphate

7- (2- (2-Thienyl) acetamido] cephalosporanic acid (2.0 g;

millimoles), 1-methyl-1 H -tetrazol-5-thiol (0.87 g;

7.5 mmol) and 3,5-dichlorophenyl dihydrogen phosphate (0.122 g, 0.5 mmol) were heated at 70 ° C overnight in 25 mL of anhydrous acetonitrile. The mixture was evaporated to a volume of 8-10 ml using a rotary evaporator. The product begins to crystallize. ^! / 40 after ₂ hours stirring dropwise over * / ₂ h 25 ml of isopropyl acetate are added dropwise in order to promote crystallization. After stirring for a further ₂ hours, the product was isolated by filtration, washed with isopropyl acetate and dried. 0.98 g (43.4% yield) is obtained. The NMR spectrum showed the product to be identical to the product of Example 5.

Example 8

Preparation of 3 - {[(1H-tetrazol-5-yl) thio] methyl} -7- [2- (2-thienyl) acetamido] -3-cephem-4-carboxylic acid in acetic acid

7- [2- (2-Thienyl) acetamido] cephalosporanic acid (2.0 g;

millimoles) and 1-methyl-1H-tetrazole-5-thiol (0.87 g;

7.5 mmol) was added to 25 ml glacial acetic acid. The reaction mixture was heated to 60 ° C and maintained at this temperature for 1 hour. The TLC on the sample taken shows only traces of the product 60. The reaction mixture was heated to 80 ° C and maintained at this temperature for 5 hours. The reaction mixture was allowed to stand overnight, the product was isolated by filtration, washed with acetic acid and dried.

The product obtained (0.71 g, 31% yield) had the same NMR spectrum as in Example 5.

-6179967

Example 9

3 - {[(4,5-dihydro-6-hydroxy-4-methyl-5-oxo-l, 2,4-triazin-3-yl) thio] methyl} -7- [2- (2- thienyl) acetamido] -3-cephem-4-carboxylic acid in acetonitrile

In an oil bath flask at 84-85 ° C, to 25 ml of anhydrous acetonitrile was added 7- [2- (2-thienyl) acetamido] cephalosporanic acid (2.0 g, 5 mmol) and 4,5-dihydro-6-hydroxy-4-methyl -5-Oxo-1,2,4-triazine-3-thiol (1.2 g, 7.5 mmol) was added. The flask was previously fitted with a condenser fitted with a drying tube containing anhydrous calcium sulfate ("Drierite"). The reaction mixture was maintained under slow magnetic stirring for 16 hours under the same conditions.

The product crystallizes from the hot solution. The reaction mixture was cooled to room temperature, filtered, washed with acetonitrile and then with acetone and dried in vacuo. 1.81 g (72.6% yield) of off-white crystals are obtained; mp 161 ° C (dec.).

On standing, further crystals precipitate out of the filtrate. These crystals were collected by filtration, washed with acetonitrile and dried. The resulting crystal (0.26 g, 10.5% yield) also had a melting point of 161 DEG C. (dec.). Thus, the total yield was 83.1%.

Example 10

Preparation of 3 - {[(5-Methyl-1,3,4-oxadiazol-2-yl) thio] methyl} -7- [2- (2-thienyl) acetamido] -3-cephem-4-carboxylic acid 1,2-dichloroethane

7- [2- (2-Thienyl) acetamido] cephalosporanic acid (2.0 g;

millimoles) and 5-methyl-1,3,4-oxadiazole-2-thiol (0.87 g;

7.5 mmol) in a 25 ml flask equipped with a magnetic stir bar and a condenser with a drying tube containing anhydrous calcium sulfate (Drierite). Immerse the flask in an oil bath of 84-85 ° C for 8 hours and then cool for two days.

A portion of the product crystallizes out, which is separated by filtration, washed with 1,2-dichloroethane and dried. The resulting material (1.10 g, 48.7% yield) was dissolved in 15 ml of acetone and the insoluble product was removed by filtration. To the solution was added dropwise 75 ml deionized water and the product was filtered off and dried. 0.61 g, m.p. 114 DEG C. (dec.).

The filtrate was evaporated to give 0.23 g of a mixture of the product and the two starting materials.

Example 11

Preparation of 3 - {[(1-Methyl-1H-tetrazol-5-yl) thio] methyl} -7- [2- (2-thienyl) acetamido] -3-cephem-4-carboxylic acid in nitromethane

To a flask of 25 ml of anhydrous nitromethane was added 7- [2- (2-thienyl) acetamido] -ephalosporanic acid (2.0 g; 5 mmol) and 1-methyl-1H-tetrazole-5-thiol (0.87 g; 7 5 mmol); immerse the flask in an oil bath of 100-101 ° C

4.5 hours and then cooled to room temperature. TLC performed indicates complete reaction.

The solvent was removed on a rotary evaporator and the residue was dissolved in 75 mL of warm ethyl acetate. The solution was filtered to remove any insolubles and extracted with 2x25 mL of 5% aqueous sodium bicarbonate. The combined extracts were shaken with 50 mL of ethyl acetate and acidified to about pH 1.0 with 70% aqueous methanesulfonic acid. The ethyl acetate layer was separated and the aqueous layer was extracted with 25 mL of ethyl acetate. The ethyl acetate layer was combined, dried over anhydrous sodium sulfate and concentrated to 25 ml on a rotary evaporator. Addition of 50 ml of diethyl ether causes the product to crystallize. The product is isolated by filtration, washed with diethyl ether and dried. 1.27 g (56.2% yield) of off-white crystals were obtained; 156-159 ° C (dec.). The product was identified by its NMR spectrum.

Example 12

Preparation of 3 - {[(Methyl-1 H -tetrazol-5-yl) thio] methyl} -7- [2- (2-thienylacetamido] -3-cephem-4-carboxylic acid in methylene chloride

7- [2- (2-Thienyl) acetamido] cephalosporanic acid (11.9 g, 30 mmol) and 1-methyl-1H-tetrazol-5-thiol (7.0 g, 60 mmol) in cyclohexane were stabilized with 300 mL of methylene chloride and placed in a 1 liter stainless steel autoclave fitted with a heater. The reaction mixture was stirred and heated at 2,856 atm at 83-86 ° C for 16 hours. The reaction mixture was then cooled to room temperature. According to TLC, the conversion is almost complete with only trace amounts of cephalosporanic acid starting material remaining. After leaving the reaction mixture at room temperature, the product crystallizes. The mixture was concentrated to 200 mL and the crystals were filtered off and washed with methylene chloride. 7.37 g (54.3% yield) of white crystals are obtained; mp 163.5-164 ° C (dec.).

Second, crystallization occurs when the filtrate is diluted with 100 mL of diethyl ether. Filtration, washing with diethyl ether and drying gave 1.40 g (10.3% yield) of a tan crystal.

The filtrate was diluted with isopropyl acetate to give a third crystalline solid: 1.18 g (8.7% yield) of crystals.

Thus, the overall yield was 73.3%.

Example 13

Preparation of 3 - {[(1-Methyl-1H-tetrazol-5-yl) thio] methyl} -7- [2- (2-thienyl) acetamido] -3-cephem-4-carboxylic acid in fluorobenzene

7- [2- (2-Thienyl) acetamido] cephalosporanic acid (2.0 g, 5.04 mmol) and 1-methyl-1H-tetrazol-5-thiol (1.2 g, 10 mmol) in 75 mL of fluorine of benzene (boiling point 85.1 ° C) in a flask equipped with a condenser containing anhydrous calcium sulfate ("Drierite"). The mixture was heated to reflux and the reaction monitored by TLC (ethyl acetate: acetic acid 7: 1). The reaction was completed in 72 hours.

The reaction mixture was cooled to room temperature and

-7179967 filtered to separate the precipitated product. The product was washed with fluox-bgnzc> 1141 and dried at 40 ° C for 5 hours under vacuum. 2.13 g (93.4%) of an off-white crystal are obtained; mp 161-162 ° C (dec.).

Example 14 _is the

3 - {[(l-Methyl-JH | ptrazol _5-yl) thio] methyl} -7- [2- (2 _thienyl) -apftamido] -3-cephem-4-karbpnsqv elcí.állításq tiofépbep

7- [2- (2-Thienyl) acetamido (19) -cephalosporan-4-one (2.0 g; 5 mL) -R 1) and 1-methyl-1H-tetrpzq-5-thiol (0.87 g;

7.5 ml) of 25 ml of thiofpnbqp is refluxed for 7 hours while the product is crystallized. The reaction mixture was cooled to room temperature, stirred for 30 minutes until crystallization occurred, and filtered. The separated product was washed with 0.5 ml of thiophene and dried in vacuo for 2 hours. 1.82 g (80.2% yield) of white crystals are obtained; 162-163 ° C (dec.). TLC indicated that the filtrate contained further product.

Example 15

3 - {[(5-Methyl-l, 3,4-thiadiazol-2-il) -tip] methyl} -7- [2- (lH-te | pyrazole: _l: yl) -acetarriido] -3 Preparation of -cgfem-4-carbpnsgy in acetyltitrile

7- (2- (1H-Tetrazol-1-yl) -acetamido] -ephaphosporanoic acid (0.76 g, 2 mmol) and 5-methyl-1,3,4- (ia (thiazol-2-thiol (0.33 g)) g; 2.5 mmol) in IQ ml of acetonitrile of purity purity was refluxed for 2 40, ηι for 40 minutes. The product crystallized out. (Vg (67% yield) identified by nmr spectra: NMR δ 2.68 (s, 3, tetrazole-C77 ₃ ),

3.72 (s, 2, 2-Cff ₂ ),

440 (q, 2, 3-C & S-, J _ab- 13 Phys),

5.12 (d, 1, C _e -Ji, J = 5 Hz),

5.38 (s, 2, -CH ₂ CONH-),

5.72 (q, 1, Cy-H, J-5 Hz, J = 9 Hz),

9.00 (s, 1, tjrazole 5-H) and

9.17 (d, 1, - CH ₂ CON / 7 -).

Example 16

Preparation of 3 - {[(1-Methyl-1 H -tetrazol-5-yl) thio] methyl} -7- (2-formyloxy-2-phenylacetamido) -3-cefcm-4-carboxylic acid, 2-diklpr etánbap

7- (2-Formyloxy-2-phenylacetamido) -cephalosporanic acid (2.17 g, 5 mmol) and 1-methyl-1H-thiazol-5-thiol (0.87 g;

7.5 mmol) of 1,2-dichloroethane-hpz (25 ml) was added and the reaction mixture was refluxed for 6 hours. After 3 and 6 hours, a sample is taken for TLC. The reaction mixture was heated for an additional 1 hour and then allowed to cool overnight.

The solvent was evaporated. Diethyl ether was added to the residue. The product, which had first become gum-like, was collected by filtration, washed with diethyl ether and dried. 1.90 g (77% yield) of product are obtained. NMR _(DMSO-d6):?

δ 3.52 (s, 2, 2-0-¾).

3.88 (s, 3, tetrazole -CH ₃ ),

4.1Q (s, 2, 3-C # ₂ S-),

4.92 (d, 1, C ₆ -H),

5.62 (q, 1, C-J-H, J = 5 Hz, 1 = 9 Hz),

6.06 (s, 1, - C //C0NH),

7.26 (s, 5, phenyl-H) and

8.28 (s, 1, -OC-H).

II

Example 17

3? - {[(l-methyl-lH-tjraZQl-5-yl) methyl} -tÍ0j -7- [5-aminocarbonyl-5- (2,4-dichlorobenzamido) valeramido] -3-cephem Preparation of -4-carboxylic acid

A) to acetonitrile, n and B) to 1,2-dichloroacetate

A) 7- [5-Carboxy-5- (2,4-dichlorobenzamido) -valeramido] -cephalosporic acid (2.9 g, 5 mmol), 1-methyl-1H-tgtrazpl-5-type (1.2 g) (IQ millimoles) and 50 mL of acetonitrile was refluxed overnight (18 hours). TLC of the withdrawn sample indicated that the reaction was about 90% complete. The reaction mixture was evaporated. the residue was suspended in ethyl acetate and filtered. 1.51 g (48.3% yield) of product are obtained.

B) 7- [5-Carboxy-5- (2,4-dichloro-benzoquino) -valeramido] -cephalosporanic acid (2.9 g, 2 mmol), 1-methylethyl-1 H -tetrazol-5-thiol (1,2) g; 10 mmol) and 50 ml of 1,2-dichloroethane was refluxed for 5.5 hours. According to TLC, the reaction was 90% complete.

The solvent was decanted from the reaction mixture and the residue was triturated with ether under reflux. The product was isolated by filtration to give 2.83 g (89.3% yield) of a tan crystal. The product was identified by its NMR spectrum. NMR (DMSO _d6):?

δ 1.78 and 2.26 (both s, adipoyl side chains),

3.66 (m, 2, 2-CV ₂ ),

3.95 (s, 3, tetrazole -CH ₂ ),

4.30 (m, 2, _3-CH2 S);

5.08 (d, 1, C ₆ -H, J = 5 Hz),

5.68 (q, 1, C-j-ff, J-5 Hz),

7.50 and 7.62 (both s, 2,4-dichlorophenyl) and 9.00 (m, 2, two -CONH).

Example 18

Preparation of 3 - {[(1-Methyl-1H-tetrazol-5-yl) thio] methyl} -7- [2- (2-thienyl) acetamido] -3-cephem-4-carboxylic acid 1,2- dichloroethane

7- [2- (2-Thienyl) acetamido] cephalosporanic acid (0.99 g;

2.5 mmol) and 1-methyl-1H-tetrazol-5-thiol (0.58 g, 5 mmol) were added to 12.5 ml 1,2-dichloroethane and the reaction mixture was refluxed. Refluxing is performed to remove the acetic acid by-product; to this end, the 1,2-dichloroethane reflux is recycled through calcium oxide.

After _^3/4 h reflux the reaction was cooled to room temperature down-8179967 and filtered. The fluffy needle crystals were washed with 10 mL of 1,2-dichloroethane and dried to give 0.77 g (68.1% yield) of product.

Concentration of the filtrate afforded another product (0.04 g, 3.5%).

The product was identified by TLC and, for the main product, by NMR. The NMR spectrum was identical to that of Example 5.

Example 19

For the preparation of 3 - {[(5-Methyl-1,3,4-thiadiazol-2-yl) thio] methyl} -7- [2- (2-thienyl) acetamido] -3-cephem-4-carboxylic acid acetonitrile

7- [2- (2-Thienyl) acetamido] cephalosporanic acid (2.0 g; 5 mmol) and 5-methyl-1,3,4-thiadiazole-2-thiol (2.64 g; 20 mmol). reflux (79 ° C) overnight in acetonitrile. TLC showed only trace amounts of cephalosporanic acid starting material. The reaction mixture was filtered to remove an oily residue and the solvent was evaporated on a rotary evaporator. The residue was crystallized from a 1: 1 mixture of isopropyl acetate / acetonitrile, separated by filtration, washed and dried in vacuo. 1.59 g (33.9% yield) of product are obtained, m.p. 166 DEG C., which is identified by IR, UV (UV), NMR and mass spectroscopy and elemental analysis.

NMR _(DMSO-d6):?

δ 2.60 (s, 3, tetrazole -CH ₃ ),

3.68 (s, 2, 2-CH ₂ ),

3.76 (s, 2, -CZ ^ CONH-),

4.38 (q, 2, 3-CH ₂ -S-, J = 13Hz),

5.10 (d, 1, C ₆ -H, J = 5 Hz),

5.70 (q, 1, C ₇ -H, J = 5 Hz, J = 9 Hz),

6.92 (d, 2, thiophene 3- and 4-H),

7.37 (t, 1, thiophene 5-H) and

9.10 (d, 1, - CH ₂ CON / 7, J = 9 Hz).

Example 20

Preparation of 3 - {[(1-Methyl-1H-tetrazol-5-yl) thio] methyl} -7- [2- (2-thienyl) acetamido] -3-cephem-4-carboxylic acid 1.1 , 2-tri chloroethane

7- [2- (2-Thienyl) acetamido] cephalosporanic acid C 2.0 g; 5 mmol) and 1-methyl-1H-tetrazole-5-thiol (1.2 g, 10 mmol) were added to 25 ml 1,1,2-trichloroethane and heated to 100-101 ° C. The reaction was monitored by TLC. When heated at 100 to 101 ° C for 4 hours with stirring, TLC shows the starting material cephalosporanic acid. The reaction mixture was cooled to room temperature with stirring. After seeding with crystalline particles, the reaction mixture is stirred overnight. The product crystallizes. The solvent was evaporated and to the product was added 25 ml 1,2-dichloroethane. The solvent was again removed by filtration, and the product was washed and dried under vacuum.

0.98 g (43.4% yield) of 3 - {[(1-methyl-1H-tetrazol-5-yl) thio] methyl} -7- [2- (2-thienyl) acetamide] -3 -cefem-4-carboxylic acid is obtained; mp 158 ° C (dec.).

Example 21

Preparation of 3 - {[(1-Methyl-1H-tetrazol-5-yl) thio] methyl} -7- [2- (2-thienyl) acetamido] -3-cephem-4-carboxylic acid ethyl ketone and

B) 1,1,2-trichloroethane

A) 7- [2- (2-Thienyl) acetamido] cephalosporanic acid (2.0 g, 5 mmol) and 1-methyl-1H-tetrazol-5-thiol (1.2 g, 10 mmol) in 25 mL of methyl ethyl ketone and refluxed for 48 hours. TLC only shows trace amounts of cephalosporanic acid starting material. The reaction mixture was washed with 2.5 g of NaHCO _{3 in} 50 ml of water. The aqueous layer was treated with 1 g of carbon and 50 mL of ethyl acetate was added. The pH was reduced to 1.6 with 4 mL of methanesulfonic acid in 30 mL of water. The ethyl acetate layer was dried over sodium sulfate and concentrated by rotary evaporation to give an oil.

The product was crystallized by dropwise addition of ether (50 mL). The product was isolated by filtration and dried in vacuo. 0.94 g (41.6% yield) of the product is identical to the product of Example 5.

B) The reaction is repeated with the above starting materials and amounts, except that 50 ml of 1,1,2-trichlorethylene are used as solvent. The reaction mixture was refluxed for 16 hours, at which time TLC showed no cephalosporanic acid starting material. The product was isolated as described in A above to give 0.47 g (20.8% yield). The NMR spectrum was identical to that of Example 5.

Example 22

Preparation of 4 - {[(1-Methyl-1H-tetrazol-5-yl) thio] methyl} -7- (2-phenylacetamido) -7-methoxy-3-cephem-4-carboxylic acid 1,2- dichloroethane

7- (2-Phenylacetamido) -7-methoxy-cephalosporanic acid (210 mg, 0.5 mmol), 1-methyl-1H-tetrazol-5-thiol (87 mg, 0.75 mmol) and 15 mL of 1, 2-Dichloroethane was added and refluxed under a nitrogen atmosphere. According to TLC, only a trace amount of cephalosporanic acid starting material is present in the reaction mixture. To the reaction mixture was again added 1-methyl-1 H -tetrazol-5-thiol (29 g, 0.25 mmol) and refluxed for 3 hours. TLC shows no appreciable change.

The reaction mixture was washed with 4x saturated sodium bicarbonate solution and the hydrocarbonate layer was washed 3x with ethyl acetate, fresh ethyl acetate was added, cooled to 0 ° C and pH 20% with hydrochloric acid.

Set to 2.2. The layers were separated and the aqueous layer was washed with ethyl acetate. The acetate layers were combined, washed with saturated sodium chloride solution, dried over magnesium sulfate, filtered and evaporated to give 199 mg (83.6% yield) of a light green foam. The product was identified by TLC and NMR. NMR (CDCl ₃ + 1d acetone-d ₆ ):

3.45 (s, 3, - OCH ₃ ),

3.55 (s, 2, 2-Cflj),

3.75 (s, 2 0CH ₂ CO-);

3.9 (s, 3, tetrazole -CH ₃ ),

4.40 (s, 2, _3-CH2 S);

-9179967

5.15 (s, 1, C ₆ -H),

7.35 (s, 5, 0),

8.00 (s, 1, - CH ₂ CONH-) and ~ 11.20 (s, 1, J = 0 Hz, - COOH).

Example 23

Preparation of 3 - {[(2-Benzothiazolyl) thio] methyl} -7- [2- (2-thienyl) acetamido] -3-cephem-4-carboxylic acid

1.2 in dichloroethane

7- [2- (2-Thienyl) acetamido] cephalosporanic acid (1.0 g;

2.5 mmol) and 2-mercaptobenzothiazole (0.625 g).

(3.75 mmol) was added to the flask, the air was evacuated under a stream of nitrogen, 25 ml of 1,2-dichloroethane was added and the reaction mixture was stirred and refluxed for 24 hours. The reaction mixture was cooled and filtered to give 1.1 g (88% yield) of an off-white solid; 190.5-191 ° C (dec.). Dry overnight in vacuo at 40 ° C and analyze. The product was identified by NMR, UV, IR, mass spectroscopy and elemental analysis.

NMR _(DMSO-d6):?

δ 3.74 (s, 2, 2-CH ₂ ),

3.80 (s, 2, -CH ₂ -CONH-);

4.58 (q, 2, 3-CH ₂ -S-, J = 13Hz),

5.14 (d, 1, C ₆ -H, J 5 Hz),

5.73 (q, 1, C ₇ - H, J 5 Hz, J = 8 Hz),

6.96; 7.43; 7.96 (both m, phenyl and thienyl rings),

9.10 (s, 1, -CH ₂ CONH-, J = 8Hz).

Example 24

3 - {[(5- / N-methylacetamido / -l, 3,4-thiadiazol-2-yl) thio} methyl-7- [2- (2-thienyl) acetamido] -3-cephem Preparation of 4-carboxylic acid in 1,2-dichloroethane

7- [2- (2-Thienyl) acetamido] cephalosporanic acid (2.0 g, 5 mmol) and 5- (N-methylacetamido) -1,3,4-thiadiazole-2-thiol (1.42 g; 7.4 mmol) was added to 50 ml of 1,2-dichloroethane and the reaction mixture was refluxed for 10 hours. The reaction mixture was cooled and filtered

It was washed with 1,2-dichloroethane and dried in vacuo. 2.06 g (78.3%) of product are obtained, m.p. 178-179 ° C, which is identified by IR, UV, NMR, mass spectroscopy and elemental analysis.

NMR (DMSO-dg):

δ 2.42 (s, 3, -COCH3),

3.74 (m, 7, 2-CH ₂ , tetrazole -CH ₃ and -CH ₂ CONH-),

4.35 (q, 2, 3-CH ₂ , J = 13 Hz),

5.10 (d, 1, C ₆ -H, J = 5 Hz),

5.70 (q, 1, C ₇ -H, J = 5 Hz, J = 9 Hz),

6.96 and 7.36 (both m, 3, thiophene H) and

9.10 (d, 1, -CH ₂ CONH-, J = 9 Hz).

Example 25

3 - (((1-Methyl-1H-tetrazol-5-yl) thio] methyl} -7- (N-tert-butoxycarbonyl-2-phenylglycylamido) -3-cephem-4 -Carboxylic acid in 1,2-dichloroethane

1.2-Dichloroethane (45 mL) was refluxed to remove the azeotropic water / solvent, then distilled to a mL remaining and the residue cooled. 7- (N-tert-Butoxycarbonyl-2-phenylglycylamido) -cephalosporanic acid (253 mg, 0.5 mmol) was added and the solution was distilled to leave a good amount of 15 mL, then the residue cool. 1-Methyl-1H-tetrazol-5-thiol (87 mg, 0.75 mmol) was added, the reaction mixture was refluxed under nitrogen and the reaction was monitored by TLC, which showed only 10 traces of starting material after 16 hours. .

The reaction mixture was washed 4 times with saturated sodium bicarbonate solution, the hydrocarbonate layers were combined and washed twice with ethyl acetate. Freshly distilled ethyl acetate is added to the washed bicarbonate phase, the solution is cooled to 0 ° C and the pH is adjusted to 2.4 with 20% hydrochloric acid. The layers are separated. The aqueous layer was washed with ethyl acetate.

The ethyl acetate layers were combined, washed with saturated sodium chloride solution, dried over magnesium sulfate, filtered and evaporated. 224 mg (80% yield) of a white foam are obtained. The product was identified by TLC and NMR. NMR showed little (10% or less) of cephalosporanic acid starting material.

The product was suspended in diethyl ether (10 ml) and rubbed with it for 1 hour. The ether is decanted, 10 ml of fresh ether are added and the mixture is rubbed for 1 hour. The ether was decanted and evaporated to dryness. The product remaining as a white powder was identified by TLC and NMR.

NMR (CDCl3):

δ 1.45 (s, 9, -COO-t-C ₄ H ₉₎

3.60 (s, 2, 2-CH ₂ ),

3.95 (s, 3, _CH3 on tetrazole);

4.30 (s, 2, _3-CH2 S);

4.90 (d, 1, J = 6 Hz, C ₆ -H),

5.40 (d, 1, J = 8Hz, 0CH--),

5.75 (q, 1, J = 4 Hz, C ₇ -H),

6.20 (d, 1, J = 8Hz, 0CH--),

NFF

7.40 (s, 5.0),

7.65 (d, 1, J = 8 Hz, - CONH) and - 9.30 (s, 1, COOH).

Example 26

3 - {[(1-Methyl-1H-tetrazol-5-yl) thio] methyl} -7- [N- (tert-butoxycarbonyl) -2- (p-hydroxyphenyl) 50-glycyl- amido] -3-cephem-4-carboxylic acid

1.2 in dichloroethane

1.2- Dichloroethane (45 mL) is refluxed with water separator and 15 mL solvent is evaporated.

The remaining 30 ml was cooled. 7- [N- (tert-Butoxycarbonyl) -2- (p-hydroxyphenyl) glycylamido] cephalosporanic acid (260.6 mg, 0.5 mmol) was added and the solution was further distilled to 15 ml. and cooling the residue. 1-Methyl-1H-tetrazol-5-thiol 60 (87 mg, 0.75 mmol) was added and the reaction mixture was heated to 65-70 ° C. Following the course of the reaction by TLC, it is found that starting material is barely present after 3 hours. The reaction mixture was worked up at the end of 4 hours.

The processing and purification is essentially the same as the previous one

-10179967. Yields were 189 mg crude product (65.5%) and 46 mg purified product (approximately 16%).

NMR (CDCl ₃ + 2d acetone d ₆ ):

δ 1.45 (s, 9, COO tert-C ₄ ffg),

3.35 (s, 2, 2-CH ₂ ),

3.85 (s, 3, _CH3 on tetrazole);

4.30 (s, 2, _3-CH2 S);

4.90 (d, 1, J = 6 Hz, C ₆ -H),

5.30 (q, 1, J = 3 Hz, C ₇ -H),

5.40 (s, 1, HO-0CH-),

5.75 (d, 1, J = 6Hz, 28),

6.40 (s, 1, COOH),

6.80 (d, 2, J = 8 Hz, 29) and f 7.90 (d, 1, - CONtf).

Example 27

Preparation of 3 - {[(1-Methyl-1H-tetrazol-5-yl) thio] methyl} -7- (2-phenoxyacetamido) -3-cephem-4-carboxylic acid in 1,2-dichloroethane

1,2-Dichloroethane (60 mL) was refluxed with water separator and 15 mL solvent was evaporated. The remaining 45 ml was cooled. 7- (2-Phenoxyacetamido) -cephalosporanic acid (406 mg, 1 mmol) was added and the solution was further distilled to 30 mL and then cooled. 1-Methyl-1H-tetrazole-5-thiol (174 mg, 1.5 mmol) was added and the reaction mixture was refluxed under nitrogen for 12 hours. A sample of the reaction mixture showed TLC (diethyl ether: acetic acid: water 1: 3: 1) as the product, excess thiol reagent, little cephalosporanic acid and an unknown component.

The reaction mixture is then worked up using the methods described in the previous two examples. TLC of the final product indicates that the product is identical to a validated sample prepared according to known prior art procedures.

NMR (CDCl3):

δ 3.65 (s, 2, 2-CH ₂ ),

3.90 (s, 3, tetrazole CH ₃ ),

4.35 (s, 2, 3-C / 7 ₂ S-),

4.65 (2, 0OCH ₂ -),

5.10 (d, 1, J = 4 Hz, C ₆ -H),. 5.90 (q, 1, J 4 Hz, C ₇ -H),

7.10 (m, 5, 0-),

7.60 (d, 1, J = 10 Hz, - CONtf -) and

-9.00 (s, 1, - COOH).

28-31. example

Preparation of 3 - {[(1-Methyl-1 H -tetrazol-5-yl) thio] methyl} -7- [2- (2-thienyl) acetamido] -3-cephem-4-carboxylic acid in fluorobenzene

3 - {[(1-Methyl-1H-tetrazol-5-yl) thio] methyl} -7- [2- (2-thienyl) acetarido] -3-cephem-4-carboxylic acid is prepared by four different reactions using a different cephalosporanic acid starting material for each of them:

28: 3- (Propionyloxymethyl) -7- [2- (2-thienyl) acetamido] -3-

-cefem-4-carboxylic acid (41 mg, 0.1 mmol),

29.: 3- (2-Methyl-propionyloxymethyl) -7- [2- (2-thienyl) -

-acetamido] -3-cephem-4-carboxylic acid (42 mg, 0.1 mmol),

30. 3- (n-Butyryloxymethyl) -7- [2- (2-thienyl) acetamido] -3-

-cefem-4-carboxylic acid (42 mg, 0.1 mmol),

31.: 3- (Cyclobutylcarbonyloxymethyl) -7- [2- (2-thienyl) -

-acetamido] -3-cephem-4-carboxylic acid (43 mg, 0.1 mmol).

In each case, the starting material was suspended in 10 ml of fluorobenzene (previously dried over a 4-A Linde aluminosilicate molecular sieve). To the reaction mixture was added 1-methyl-1 H -tetrazol-5-thiol (18 mg, 0.15 mmol) and refluxed for about 24 hours. For each reaction, the product precipitates, which is filtered off and dried. TLC of the product (in ethyl acetate / acetic acid) indicated that the substitution was complete. The NMR of the products is identical to the authentic sample produced by displacement in aqueous medium.

Example 32

Preparation of 3 - {[(3-Methyl-1,2,4-oxadiazol-5-yl) thio] methyl} -7- [2- (2-thienyl) acetamido] -3-cephem-4-carboxylic acid 1,1,2-trichloroethane

7- [2- (2-Thienyl) acetamido] cephalosporanic acid 40 ml

Suspend in 1,1,2-trichloroethane and add 3-methyl-1,2,4-oxadiazole-5-thiol (116 mg, 1 mmol). The reaction mixture was heated in an oil bath at 113 ° C for about 3 hours, then allowed to cool overnight and evaporated. To the residual oil was added ethyl acetate and a saturated NaHCO3 solution. The ethyl acetate layer was washed again with saturated NaHCO ₃ solution, the aqueous layers were combined and extracted again with ethyl acetate. The aqueous phase was shaken with fresh ethyl acetate, cooled in an ice bath and adjusted to pH 2.5 with 20% hydrochloric acid. The layers were separated and the aqueous layers re-extracted. Wash the combined ethyl acetate layers were washed with brine, dried over _MgSO4, filtered and evaporated. To the residue was added 1,1,2-trichloroethane to give 220 mg of solid (48% yield). The product was identified by NMR, IR, UV spectroscopy and bioautogram.

NMR _(DMSO-d6):?

δ 2.35 (s, 3, CH _{3 on} the oxadiazolone),

3.70 (q, 2, J = 18 Hz, 2-CH _2);

3.80 (s, 2, -CH ₂ CONH-),

4.40 (q, 2, J = 14Hz, _3-CH2 S);

5.10 (d, 1, J = 5 Hz, C ₆ -H),

5.60 (q, 1, J = 4 Hz, C ₇ -H),

6.90; 7.30 (t, d, 3, thiophene H) and

9.10 (d, 1, J = 8Hz, -CH ₂ CONtt-).

Example 33

Preparation of 3 - {[(1H-1,3,4-triazol-5-yl) thio] methyl} -7- [2- (2-thienyl) acetamido] -3-cephem-4-carboxylic acid

1,1,2-trichloroethane

7-6- (2- (2-Thienyl) acetamido) cephalosporanic acid 396 mg;

millimoles) were suspended in 30 ml 1,1,2-trichloroethane and 1 H -1,3,4-triazol-5-thiol (100 mg; 1 millimole) was added. The reaction mixture was heated to 105 ° C. The product precipitates in 30 minutes. The reaction mixture was heated at 100 ° C for a further 6 hours and then cooled to room temperature

-11179967 stool. The product was collected by filtration and washed with 1,1,2-trichloroethane. 380 mg (87% yield) of product are obtained which are identified by IR, NMR, UV spectroscopy and bioautogram. The bioautogram also identifies cephalosporanic acid starting material, which is 6.8% based on HPLC.

NMR _(DMSO-d6):?

3.70 (s, 2, 2-C H ₂ ),

3.80 (s, 2, - CH ₂ - CONH -), 10

4.20 (q, 2, J = 5 Hz, C-3/7 S _2),

5.10 (d, 1, J = 5 Hz, C ₆ -H),

5.70 (q, 1, J = 4 Hz, C ₇ -H), 7.00; 7.40 (t, d, 3, thiophene H),

8.45 (s, 3, thiazole N77) and 15

9.13 (d, 1, J = 8Hz, -CH ₂ CONtf-).

NMR _(DMSO-d6):?

1.97 (s, 3, CT / JCONH--),

3.70 (s, 2, 2-CN ₂ ),

4.00 (s, 3, CH ₃ a tetrazolone), 4.35 (s, 2, 3-CH ₂ S-),

5.10 (d, 1, J = 5 Hz, C ₆ -H),

5.70 (q, 1, J = 4 Hz, _C7 --H) and

8.80 (d, 1, J = 8 Hz, - CH ₂ CON / 7).

Example 36

Preparation of 3 - {[(3-Methyl-1,2,4-thiadiazol-5-yl) thio] methyl} -7- [2- (2-thienyl) acetamido] -3-cephem-4-carboxylic acid 1,2-dichloroethane

Example 34

Preparation of 3 - {[(1-Methyl-1H-tetrazol-5-yl) thio] methyl} -7- (2-phenylacetamido) -3-cephem-4-carboxylic acid in chloroform

7- (2-Phenylacetamido) -cephalosporanic acid (195 mg, 25 0.5 mmol) was suspended in chloroform (75 mL) and 1-methyl-1H-tetrazol-5-thiol (75 mg, 0.65 mmol) was added. The reaction mixture was heated in an oil bath at 80-85 ° C for 3 hours and 60 ml was distilled off. TLC showed only a small amount of product, so 20 mL of 1,2-30 dichloroethane was added and heating was continued overnight. The following morning, the TLC sample from the reaction mixture showed little cephalosporanic acid starting material. After a total reaction time of 26 hours, the reaction mixture was cooled to room temperature. The product was subjected to the procedure of Example 35 to obtain 80 mg of purified material (35% yield). The product was identified by NMR, IR, UV spectroscopy and bioautogram.

NMR _(DMSO-d6): 40

3.60 (s, 2, 2-C / 7 ₂ ),

3.70 (s, 2, -C / 7 ₂ CONH-),

4.00 (s, 3, Cff _{3 on} the tetrazolone),

4.30 (s, 2, 3-C / 7 ₂ S-),

5.10 (d, 1, J = 5 Hz, C ₆ -H), 45

5.70 (q, 1, J = 4 Hz, C ₇ -H),

7.30 (s, 5.0) and

9.13 (s, 1, J = 8Hz, --CH2 CONONF -).

Example 35

Preparation of 3 - {[(1-Methyl-1H-tetrazol-5-yl) thio] methyl} -7-acetamido-3-cephem-4-carboxylic acid

In 1,2-dichloroethane 55

7-Acetamidocephalosporanic acid (314 mg, 1 mmol) and 1-methyl-1H-tetrazol-5-thiol (98 mg, 0.65 mmol) were mixed with 70 mL of 1,2-dichloroethane. 50 ml of the reaction mixture are distilled off and the residue is refluxed for about 24 hours.

The reaction mixture was worked up as described in Example 32 to give 120 mg (32% yield) of product which was identified by NMR, IR, UV spectroscopy and bioautogram. 65

7- [2- (2-Thienyl) acetamido] cephalosporanic acid (200 mg; 0.5 mmol) and 3-methyl-1,2,4-thiadiazole-5-thiol (85 mg; 0.65 mmol) It was stirred in 1,2-dichloroethane (ml) and heated to 95 ° C in an oil bath. The bath temperature was maintained at 90 ° C for 19 hours. The reaction flask was then removed from the bath, allowed to cool, and cooled for 1 day. Add 3 volumes of ethyl acetate and wash with 2x50 mL of saturated sodium bicarbonate solution. The combined aqueous washings were extracted with ethyl acetate, shaken with fresh ethyl acetate, and cooled in an ice bath. The pH is adjusted to 2.0. The layers were separated and the aqueous layer was re-extracted with ethyl acetate. The combined ethyl acetate extracts were washed with saturated brine, dried over magnesium sulfate, filtered and evaporated. The foam (252 mg) was crystallized from acetone / diethyl ether to give 153 mg (65%). The product was identified by NMR, UV, IR spectroscopy and bioautogram.

NMR _(DMSO-d6):?

3.70 (s, 2, C-CH ₂ ),

3.80 (s, 2, - CH ₂ CONH),

4.50 (q, 2, J = 15 Hz, 3-C77 ₂ -S-),

5.10 (d, 1, J = 5 Hz, C ₆ -H),

5.70 (q, 1, J = 4 Hz, C ₇ -H \

6.96; 7.38 (t, d, 3, thiophene H) and

9.13 (d, 1, J = 8 Hz, --CH2 CONONF).

Example 37

Preparation of 3 - {[(2-pyrimidinyl) thio] methyl} -7- [2- (2-thienyl) acetamido] -3-cephem-4-carboxylic acid in acetomtril

7- [2- (2-Thienyl) acetamido] cephalosporanic acid (2.0 g; · 5 mmol) and 2-mercaptopyrimidine (0.62 g; 5.5 mmol) in 25 mL of anhydrous acetonitrile overnight (16 hours) with stirring and reflux. TLC indicates complete conversion to product. The reaction mixture was cooled to room temperature and the product was isolated by filtration, washed with 50 ml of acetonitrile and dried in vacuo at 50 ° C for 4 hours. 1.86 g of an off-white crystal are obtained (83% yield); mp 217 ° C (dec.). Nothing shows the NMR spectrum (in _DMSO-d6) a starting material. The product was identified by IR, UV and NMR spectroscopy.

-12179967

NMR (DMSO-dg):

δ 3.55 (q, 2, 2-CH ₂ , J = 18 Hz),

3.74 (s, 2, - CW ₂ CONH),

4.28 (9, 2, 3-CH ₂ S -, 3 = 13 Hz),

5.00 (d, 1, Cg-H, J = 5 Hz),

5.64 (q, 1, C ₇ -H, J = 5 Hz, J = 9 Hz),

7.08 (m) and 8.52 (d) (6, thiophene and pyrimidine H) and

9.00 (d, 1, -CH ₂ CON7 / -, J = 9 Hz).

Example 38

Preparation of 3 - {[(2-pyrimidinyl) thio] methyl} -7- [2- (2-thienyl) acetamido] -3-cephem-4-carboxylic acid in acetic acid

7- [2- (2-Thienyl) acetamido] cephalosporanic acid (2.0 g, 5 mmol), 2-mercaptopyrimidine (0.6 g, 5.4 mmol) and sodium acetate (0.41 g) (5 mmol) in 25 ml of glacial acetic acid was heated at 85 ° C for 4 hours. The product crystallizes during the reaction. The reaction mixture was cooled to room temperature and the product was isolated by filtration, washed with acetic acid and dried. 1.58 g (70.5% yield) of white crystals are obtained; mp 218 ° C (dec.). The product was identified by IR, UV, NMR spectroscopy and elemental analysis. The NMR spectrum was identical to that of Example 37.

Example 39

Preparation of 3 - {[(1-Methyl-1 H -tetrazol-5-yl) thio] methyl} -7- [2- (2-thienyl) acetamido] cephalosporanic acid in acetic acid

7- [2- (2-Thienyl) acetamido] cephalosporanic acid (2.0 g, 5 mmol), 1-methyl-1H-tetrazol-5-thiol (0.81 g, 7 mmol) and sodium acetate ( 0.41 g (5 mmol) in 25 ml glacial acetic acid was heated at 75-77 ° C for 8 hours. Each hour, samples are taken for TLC and at 8 hours, the reaction is complete. The reaction mixture was cooled to 40-45 ° C and the acetic acid was removed in vacuo. To the residue were added ethyl acetate (50 mL) and water (50 mL). The aqueous layer was acidified to pH 1.5 with sulfuric acid In. The ethyl acetate layer was separated, dried over anhydrous sodium sulfate and the ethyl acetate removed on a rotary evaporator. The light oily residue was dissolved in ethanol (50 mL) and dicyclohexylamine (10.2 mmol) dissolved in ethanol (10 mL). The product precipitated as dicyclohexylamine salt almost immediately, but was stirred for a further 15 minutes, then filtered off and washed with ethanol. After drying, 1.2 g (37.8% yield) of product are obtained, m.p. 185-186 ° C. The IR, NMR and UV spectra are identical to those of the previously prepared samples of the product.

Example 40

Preparation of 3 - {[(1-Benzyl-1H-tetrazol-5-yl) thio] methyl) -7- [2- (2-thienyl) acetamido] -3-cephem-4-carboxylic acid

1,2-dichloroethane

7- [2- (2-Thienyl) acetamido] cephalosporanic acid (2.0 g, 5 million mol) and 1-benzyl-1H-tetrazol-5-thiol (1.44 g;

7.5 mmol) was added to 25 ml of 1,2-dichloroethane, the reaction mixture was heated in an oil bath to 85 ° C and kept under stirring overnight at this temperature. TLC does not detect cephalosporanic acid starting material. Removal of the solvent on a rotary evaporator leaves foam. Methanol (25 mL) was added to the foam and the mixture was heated on a steam bath until the sample dissolved. The solvent was evaporated on a rotary evaporator and the product crystallized. It is stirred for 15 minutes, filtered off, washed with methanol and dried in vacuo. 1.6 g (60.6% of theory) of product are obtained with a melting point of 171-1711.5 ° C.

Example 41

Preparation of 3-amidinothiomethyl-7- [2- (2-thienyl) acetamido] -3-cephem-4-carboxylic acid in acetonitrile

7- [2- (2-Thienyl) acetamido] cephalosporanic acid (3.12 g;

and thiourea (912 mg; 12 mmol) in 15 ml of acetonitrile (previously dried over a 4-A Linde aluminosilicate molecular sieve) was heated at 87 ° C for 24 hours with stirring. Within 1 hour, a precipitate began to precipitate out of the reaction mixture.

At the end of 24 hours, the product is separated from the hot reaction mixture by filtration. After drying, 2.5 g (76% yield) of product are obtained.

Elementary analysis:

C H N calculated 43,68 3.91 13.58 Found 43.50 4.03 13.29

The product was identified by its NMR spectrum.

Example 42

Preparation of 3-Benzoylthiomethyl-7- [2- (2-thienyl) acetamido] -3-cephem-4-carboxylic acid in isopropyl acetate

7- [2- (2-Thienyl) acetamido] cephalosporanic acid (1.0 g;

2.5 mmol) and thiobenzoic acid (510 mg; 3.7 mmol) are suspended in 20 ml of anhydrous isopropyl acetate and the suspension is refluxed for 31 hours. During this time all the reagents are dissolved · The solvent is evaporated off under vacuum. The residue, according to TLC, consists of a 60:40 mixture of product and unreacted starting material.

Prepare by layer chromatography on silica gel plates (acetone: acetic acid 16: 1) to obtain a purified sample of the product.

NMR (DMSO-d 6 / D ₂ O) δ 3.50 (ABq, 2H, 2-Cf ₂ , J = 6 Hz, J = 19 Hz),

3.82 (s, 2H, - CH ₂ CONH-),

4.20 (d, 2H, C-3/7 ₂ S-, J = 5 Hz);

5.03 (d, 1H, C ₆ -H, J = 5Hz),

5.69 (d, 1H, _C7-H, 3 = 5 Hz);

6.89-8.0 (m, 8H, aromatic).

Example 43

Preparation of 3 - [(phenylthio) methyl] -7- [2- (2-thienyl) acetamido] -3-cephem-4-carboxylic acid in 1,2-dichloroethane

7- [2- (2-Thienyl) acetamido] cephalosporanic acid (2.0 g;

million moles) and benzolthiol (0.75 mL, 7.5 mmol) were added

-13179967 ml to 1,2-dichloroethane. The reaction mixture is refluxed overnight; No residual starting material was detected by TLC.

The solvent was evaporated on a rotary evaporator and the residue was partially dissolved by addition of ethanol (25 ml) and heating. Methanol (25 mL) was added to dissolve the residue and heated. The resulting solution was filtered through cotton wool, treated with 2.0 g of carbon, stirred for 5 minutes and filtered through filter paper. The solvent was evaporated on a rotary evaporator. The residual NMR spectrum showed little thiol starting material.

To the residue is added 25 ml of isopropyl acetate, the suspension is filtered and cooled overnight. The filtrate was diluted with ethyl acetate (25 mL) and washed with dilute sodium bicarbonate (25 mL) to remove the thiol. The ethyl acetate layer was added to 25 mL of water and the pH adjusted to 1.4 with sulfuric acid (8.5). The ethyl acetate layer was dried over magnesium sulfate and filtered. Concentration on a rotary evaporator gave 0.92 g (19.4% yield) of a foam.

NMR _(DMSO-d6) δ 3.58 (m, 2, _2-CH2);

3.75 (s, 2, -CH ₂ CONH-),

4.12 (q, 2, 3-CH ₂ S-J = 13 Hz),

5.08 (d, 1, C ₆ -H, J = 5 Hz),

5.66 (q, 1, C ₇ -H, J = 5 Hz, J = 9 Hz),

7.15 (m, 8, thiophene H and phenyl H) and

9.10 (d, 1.1, - CH 2 CONH -, J = 9 Hz).

Example 44

Preparation of 3 - {[(5-Methyl-1,3,4-thiadiazol-2-yl) thio] methyl} -7- (2-phenylacetamido) -3-cephem-4-carboxylic acid 1,2- dichloroethane

7- (2-Phenylacetamido) -cephalosporanic acid (1.95 g, 5 mmol) and 5-methyl-1,3,4-thiadiazole-2-thiol (0.99 g;

7.5 mmol) in 25 ml 1,2-dichloroethane was refluxed for 16 hours. TLC only shows trace amounts of cephalosporanic acid starting material. The product crystallizes during the reaction.

The reaction mixture was cooled to 0-5 ° C and filtered, and the product was washed with cold 1,2-dichloroethane. Drying in vacuo gave 1.82 g (78.8% yield) of product, mp 171-172 ° C.

NMR (DMSO-dg) δ 2.70 (s, 3, thiazolidyl-CH ₃ ),

3.58 (s, 2, -CH ₂ CONH-),

3.70 (s broad, 2, 2-CH ₂ ),

4.40 (q, 2, 3-CH ₂ S-, J = 13 Hz),

5.12 (d, 1, C ₆ -H, J = 5 Hz),

5.77 (q, 1, C ₇ -ff, 1 = 5 Hz, J = 9 Hz),

7.28 (s, 5, phenyl ff) and

9.08 (d, 1, -CH ₂ CONH-, J = 9 Hz).

Example 45

Preparation of 3 - [(methylthio) methyl] -7- [2- (2-thienyl) acetamido] -3-cephem-4-carboxylic acid in methylene chloride

7- [2- (2-Thienyl) acetamido] cephalosporanic acid (24 g, 60 mmol), 7.0 mL of methanethiol and 600 mL of methylene chloride were heated in a bomb tube at 84-86 ° C for 18 hours with stirring. The reaction mixture was then cooled to room temperature. The remaining insoluble material is filtered off and the filtrate is evaporated. The residue was taken up in ethyl acetate and filtered. About 150 mL of water was added to the filtrate and stirred; The pH is adjusted to 5.5 by the dropwise addition of sodium hydroxide solution. The aqueous phase was separated and concentrated to a volume of about 75 mL, diluted to 700 mL with water and glacial acetic acid was added to pH 3.8. An amorphous solid precipitates, which is stirred in an ice bath for 3 hours and then cooled overnight.

The solid was filtered off. To the filtrate was added 100 mL of ethyl acetate and the pH was adjusted to 2.0 with concentrated hydrochloric acid. The organic layer was separated and extracted with ethyl acetate (1 x 100 mL, 15 x 150 mL). The combined organic layers were dried over magnesium sulfate and the solvent was evaporated to give an amber foam. The latter is dissolved in 50 ml of ethyl acetate, inoculated with the precipitate of the precipitated product and kept overnight in a refrigerator. The resulting crystals were collected by filtration, washed with cold isopropyl acetate and dried in vacuo at 50 ° C to give 3.2 g (14% yield) of product, identified by NMR.

NMR δ 2.00 (s, 3, -3-CH ₂ SCH ₃ ),

3.74 (m, 3-CH ₂ S-, 2-CH ₂ , and -CH ₂ CONH -),

5.14 (d, 1, C ₆ -H, 1 = 5 Hz),

6.64 (q, 1, C ₇ H, 1 = 5 Hz, J = 9 Hz),

7.15 (m, 3, thiophene ff) and

9.12 (d, 1, - CH ₂ CONH-, J = 9 Hz).

Example 46

Preparation of the sodium salt of 7- (2-Formyloxy-2-phenylacetamido) -3 - {[(1-methyl-1H-tetrazol-5-yl) thio] methyl} -3-cephem-4-carboxylic acid benzene

7- (2-Formyloxy-3-phenylacetamido) -cephalosporanic acid (2.17 g, 4.65 mmol) and 1-methyl-1H-tetrazol-5-thiol (0.87 g, 7.5 mmol) 25 ml of benzene are refluxed (about 80 ° C) for 12 hours. During the reaction, a soft plastic-like substance is deposited in the flask, which solidifies in the glass upon cooling.

According to TLC, the solution contains only traces of product and the plastic mainly consists of the desired product and little deformylated product. After addition of acetone, the reaction mixture was evaporated. The residual foam was dissolved in 50 ml of anhydrous acetone and dissolved in acetone

Treat with 1.25 g (7.5 mmol) of sodium 2-ethylhexanoate. The product crystallizes in the form of its sodium salt. After 1 hour, the product is filtered off, washed with 20 ml of acetone and dried. 1.38 g (58% yield) of product are ka55 red, identified by TLC and NMR spectroscopy.

NMR (D ₂ O) δ 3.48 (q, 2, 2-CH ₂ , J = 18 Hz),

4.00 (s, 3, tetrazolyl-C / 7 ₃ ),

4.10 (m, 2, 3 — Cff ₂ S—),

5.05 (d, 1, C ₆ -H, 1 = 5 Hz),

5.70 (d, 1, C ₇ - H, 1 = 5 Hz),

6.24 (s, 1, -CHCONH),

7.50 (m, 5, phenyl H) and

8.33 (s, 1, 30).

-14179967

Example 47

Preparation of 7- (2-Formyloxy-2-phenylacetamido) -3 - {[(1-methyl-1H-tetrazol-1-yl) thio] methyl} -3-cephem-4-carboxylic acid in methane 5

7- (2-Formyloxy-2-phenylacetamido) -cephalosporanic acid (2.17 g, 4.65 mmol) and 1-methyl-1H-tetrazol-5-thiol (0.87 g, 7.5 mmol) ) In 25 ml of tetrachloromethane at reflux (about 77 ° C) for 12 hours. The reaction mixture was cooled to room temperature and the supernatant was decanted from the semi-solid. 25 ml of 1,2-dichloroethane are added to the latter while heating. The product crystallizes. After cooling to room temperature and stirring for about 1 hour, the product was isolated by filtration, washed with 10 ml of 1,2-dichloroethane and dried overnight at 45 ° C in vacuo. 1.54 g (65% yield) of white crystals are obtained which are identified by NMR spectroscopy and TLC.

The latter also shows little deformylated product. The NMR spectrum was identical to that of Example 16.

Example 48

3-methyl-l, 2,4-oxadiazol-5-thiol Preparation

Acetamide oxime (30 g, 0.4 mol), carbon disulfide (100 mL, 1.66 mol) and triethylamine (56 mL, 0.4 mol) were stirred in 1 L of pyridine. A stream of nitrogen was bubbled through the carbon disulphide solution and passed over the reaction mixture. The mixture was heated in an oil bath at 70 ° C for 3 days and then evaporated to an oil to which ethyl acetate and saturated sodium bicarbonate solution were added. The layers were separated and the organic layer was washed again with saturated sodium bicarbonate solution. The combined sodium bicarbonate washings were extracted with ethyl acetate and the ethyl acetate extracts discarded. To the aqueous phase was added fresh ethyl acetate and cooled in an ice bath. The pH is adjusted to 2.5 with 20% hydrochloric acid and the solution is saturated with sodium chloride. It was then extracted with ethyl acetate, and the extract was washed with saturated sodium chloride solution and dried over anhydrous magnesium sulfate. The solution was filtered and concentrated to half its original volume. An equal amount of tetrachloromethane was added and evaporation continued until the product crystallized. 24.8 g (52% yield) of product are obtained.

Example 49

Preparation of 3 - {[(3-methyl-1,2,4-oxadiazol-5-yl) thio] methyl} -7- [2- (2-thienyl) acetamido] -3-cephem-4-carboxylic acid 1,2-dichloroethane

7- [2- (2-Thienyl) acetamido] -3- (carbamoyloxymethyl) -3-cephem-4-carboxylic acid (100 mg, 0.25 mmol) in 25 mL

It is suspended in 1,2-dichloroethane and 1,2,4-oxadiazol-5-thiol (35 mg, 0.30 mmol) is added with stirring. The reaction mixture was heated in an oil bath at 110 ° C and 5 mL of solvent was distilled off to remove traces of water. The oil bath temperature was then reduced to 90-95 ° C and the mixture was maintained at this temperature for 19 hours and then cooled to room temperature. The insoluble material was removed by filtration and washed with 1,2-dichloroethane and diethyl ether. TLC was found to consist of unreacted starting material containing only traces of product.

The filtrate was evaporated and to the residual oil was added sodium bicarbonate solution and ethyl acetate. After shaking, fresh ethyl acetate was added to the separated aqueous layer, cooled in an ice bath, and the pH was adjusted to 2.5 with 20% hydrochloric acid. The organic layer was removed. The aqueous layer was extracted with fresh ethyl acetate and the combined ethyl acetate solutions were washed with saturated sodium chloride solution, dried over magnesium sulfate, filtered and evaporated. The residual oil was crystallized from 15: hexane: diethyl ether to give 13 mg (11.5% yield). The product was identified by NMR and IR; these spectra are identical to those obtained in Example 32.

Example 50

3 - {[(l-methyl-lH-tetrazol-5-yl) thio] methyl} -7- [2- (tert-butoxycarbonylamino) -2-phenylacetamido] -3-cephem-4- preparation of carboxylic acid in nitromethane

7- [2- (tert-Butoxycarbonyl) -2-phenylacetamido] -cephalosporanic acid (490 mg, 1 mmol) and 1-methyl-1H-tetrazole-5-thiol (145 mg, 1.25 mmol) Heat in anhydrous nitro30 methane (85 ml) at 85-90 ° C under nitrogen for 8 hours. It indicates TLC product, excess thiol reagent and decarboxylated product but does not detect cefolosporanic acid starting material. Nitromethane was removed by evaporation. The residual orange foam was dissolved in 10 mL of saturated sodium bicarbonate solution, water (20 mL) was added and the mixture was washed with ethyl acetate until the washings remained clear. The ethyl acetate washings were combined, water (20 mL) was added, cooled to 0 ° C, and the pH was adjusted to 2.2 with 20% HCl. The layers were separated and the aqueous layer was washed with ethyl acetate. The ethyl acetate layers were combined, washed with saturated sodium chloride solution, dried over magnesium sulfate, filtered and evaporated. 455 mg (83% yield) of a brown foam are obtained, which according to TLC 45 consists of product, traces of thiol and traces of decarboxylated product.

The product (455 mg, 0.833 mmol) was dissolved in ethyl acetate (7 mL) and lithium acetate (0.833 mL) was added dropwise with stirring. The brown tap of the lithium salt precipitates in the form of a dude. It was separated by filtration, washed with ethyl acetate and dried overnight in vacuo at room temperature. 368 mg (80% yield) of product are obtained which is identified by TLC, NMR spectroscopy, bioautogram, elemental analysis, IR and UV spectroscopy. TLC shows traces of decarboxylated material.

NMR (CDCl ₃ ) δ 1.4 (s, 9, - COO - C ₄ H ₉ ),

3.6 (s, 2, _2-CH2 \ 3.85 (s, 3, tetrazol CW _3), 4.3 (s, 2, 3 ₂ CFF-S-)

4.44 and 4.45 (2s, 1, 0C7T-), 4.9 (d, 1, J = 6Hz, C ₆ -H), 5.8 (q, 1, J = 6Hz, C ₇ -H), 7.35 (s, 5, 0),

-15179967

7.36 (s, 5.0),

8.2 and 7.8 (2d, 1, J = 9 Hz, - CONH--) and

9.3 (s, l, COO //).

Example 51

3 - {[(l-methyl-lH-tetrazol-5-yl) thio] methyl} -7- {N - [(l, 3-dimethyl-ureido) -karbpnil] -2-phenyl-glycyl-arnidp } Preparation of -3-cephem-4-carboxylic acid in nitromethane

7- {N - [(1,3-Dimethyl-ureido) -carbonyl] -2-phenylglycyl-amidp} -cephalosporanic acid (130 mg, 0.25 mmol) was added to 5 ml of n-trimethane-methane suspension and 1-methyl-1 1H-Tetrazpl-5-thiol (43.5 mg, 0.375 mmol). The slurry was heated at 85 ° C for 12 hours under nitrogen and then cooled to room temperature over the weekend. The reaction mixture was then filtered and the non-blocked material was washed with a small amount of nitromethane and dried in a vacuum oven at 35 ° C. The product (83 mg, 58% yield) was identified by TLC, NMR, IR, UV spectroscopy, elemental analysis and bioautogram. NMR _(DMSO-d6) δ 2.65 ((1, 3, J = 4Hz, - CONHC77 _3);

3.15 (s, 3, CONHCJ / jCO),

3.6 (s, 2, 2 CÉ ₂ \

3.9 (s, 3, tetrazole CHJ,

4.3 (s, 2, 3 — Cfi ₂ S—),

5.0 (d, 1, J 5 Hz, C ₆ H),

5.5 (d, 1, J = 7Hz, 31), (q, I, C ₇ H),

5.8 (q, 1, - CON // CH 3),

7.4 (s, 5.0), '

9.3 (d, 1, J = 8 Hz, OCHCONW--) and (d, 1, j = 8 Hz, 32).

Example 52

Preparation of 3 - {[(1-Methyl-1 H -tetra-1 P-5-yl) thio] methyl} -7- [2- (2-thienyl) acetaridol-3-cephem-4-carbpnsay apptpnjtrilbgn, tetrabutyl] by adding -ammóniprn-jodjd

7 - [(2-Thiepyl) acetamido] -ephephalosporanic acid (2.0 g; 5 mmol), 1- (methyl-1 H -tetrazol-5-thiol (1.2 g, 10 mmol) and tetrabutylammonium iodide (0.2 g) was heated to reflux in 25 mL of 45 mL of anhydrous acetonitrile and refluxed for 8 h. The product precipitated out of the alkali of the light kernel-like crystals was filtered off, dried over isopropyl acetate and dried to give 1.30 g (57.5% yield) of product having the same NMR spectrum as in Example 5.

Example 53

Preparation of 3 - {[(1-Methyl-1H-tetrazol-5-yl) thio] methyl} -7- [2- (2-thienyl) acetamido] -3-cephem-4-carboxylic acid 1,2- dichloroethane with addition of tetrabutylammonium iodide

The reaction described in Example 52 was repeated, except that 1,2-dichloroethane was used as solvent. Addition of dicyclohexylamine (2 mL) gave the product as the dicyclohexylamine salt (1.55 g, 48.9% yield). The NMR spectrum was identical to that of Example 1.

Example 54

Preparation of 3 - [(phenylthio) methyl] -7- [2- (2-thienyl) -acetarmo] -3-cephpn) 10 -4-carboxylic acid 1,2-dichloroethane-1-naphthyl-5 - (methyl-type) -1H-tetrazole

7- [2- (2-Thienyl) acetamido] cephalosporanic acid (2.0 g;

of benzylthiol (0.75 mL; 7.5 mL) and 0.65 g of 15-methyl-5- (methylthio) -1H-tetrazole in 15 mL of 1,2-dichloroethane was heated to reflux. By TLC, it is found that the 14 µg substrate is completely complete. The solvent was removed on a rotary evaporator and the residue q was repeatedly extracted with diethyl ether. After complete removal of the product, 1.66 g of a tan solid was obtained. (74% yield). The IR and NMR spectra were similar to those obtained in the previous examples.

Example 55

Preparation of 3 - {[(1-Methyl-1 H -tetrazol-5-yl) -type] methyl} -7- [2- (2-thienyl) acetamido] -3-cephem-4-carboxylic acid in isopropanol

7- [2- (2-Thienyl) -acetamoyl (o) -ephephosphatase (2.0 g; millimoles) and 1-methyl-1H-ether-3β-5-ol (0.87 g, 7.5 mmol) ) in 25 ml izopropanplban lpmbjkt> Kunena 35 of [the flask in dry-kglejqni szulfátpt ( "Dricrite) equipped with a condenser with a drying tube containing]. the reaction követjük- 82-83 ^c C. cefalosppránsavnak only half the TLC under 40 hours responds.

Example 56

Preparation of 3 - [(2-oxazolylthio) methyl] -7- [2- (1H-tetrazin-1-yl) acetamido] -3-cephem-4-carboxylic acid in pyromethane

7- [2- (1H-1'-tetrazol-1-yl) -acetamido] -cephalosporanic acid (0.38 g, 1 mmol) and 2-oxazolyl thiol (0.11 g, 1.1 mmol) in 5 mL of nitro- in methane in an oil bath at 90-91 ° C. A dry nitrogen atmosphere is maintained over the reaction mixture. After 20 minutes, all reagents dissolve and after 35 minutes the product begins to crystallize. After 6 hours, the req.c. mixture is cooled to room temperature and the product is filtered off, 7 m. it was washed with nitromethane, air-dried for 55 hours and then vacuum dried for 1 hour at 40 ° C.

Obtained as an off-white crystal (0.36 g, 85.7%); mp 196 ° C (dec.). The NMR spectrum consists of 70% desired product and 30% cephalosporanic acid starting material.

The product was recrystallized from 5 ml of DMSO-d ₆ and 10 ml of water, filtered off, washed with 5 ml of water: pMSO-d ₆ 2: 1 and air-dried at 0 ° C for 6 hours under vacuum at 0.26 g. 87% of the desired product and 13% of the 65 cephalosporanic acid starting materials were obtained. The recrystallized 16

-16179967 was repeated with 3 ml of DMSO-d ₆ and 6 ml of water and stirred for 1 hour to give 0.21 mg of product, identified deep by NMR; according to this spectrum, the cephalosporanic acid starting material was reduced to 5%.

NMR (DMSO-d6 δ 3.75 (s, 2, 2-0¾).

4.34 (q, 1, 3 — C # ₂ S—, 3 = 14 Hz),

5.16 (d, 1.06 - //, 3 = 5 ten),

5.76 (<j, 1, C ₇ -H, J = 5 Hz, J = 9 Hz),

5.44 (s, 2, -C ^CÖNfa—),

7.28 (s, 1, oxazole t ₄ -77);

8.14 (s, 1, oxazole C ₅ -77),

9.37 (s, 1, tetrazole II) and

9.53 (d, 1, - CON / 7 -, J = 9 Hz).

Example 57

Preparation of 3 - [(2-oxazolthio) methyl] -7- (2-formyloxy-2-phenylacetamido) -3-cephem-4-carboxylic acid in 1,2-dichloroethane

7- (2-Formyloxy-2-phenylacetamido) -cephalosporanic acid) methylene chloride solvate (0.52 g, 1 mmol) and 2-oxazolethiol (0.12 g, 1.1 mmol) in 20 mL of Dissolve in 2-dichloroethane, reflux for about 16 hours, then cool to room temperature. TLC shows complete conversion to product. The reaction mixture was evaporated to 10 ml on a rotary evaporator; on standing, slightly gelatinous crystals are formed, deep filtered, washed with 1,2-dichloroethane and dried. 12 g of a gray solid are obtained.

The solvent was completely removed from the filtrate to give 0.45 g of a pale yellow foam, which was triturated with 25 ml of diethyl ether, filtered, washed with diethyl ether and dried. 0.21 g of a light yellow powder are obtained. The product was identified by NMR spectroscopy:

δ 3.56 (m, 2, 2-CT7 ₂ ),

4.24 (q, 2, 3 - C / J ₂ S -, J = 13 Hz),

5.00 (d, 1, C ₆ - H, J = 5 Hz),

5.70 (q, 1, C ₇ -H, J = 5 Hz, J = 9 Hz),

6.14 (s, 33),

7.25 (s, 1, oxazole C ₄ -H),

7.45 (m, 5, phenyl 77),

8.12 (s, 1, oxazole ₅ -77 C),

8.35 (s, 1, - CHO) and

9.40 (d, 1, - CON77 -, J = 9 Hz).

Example 58

Preparation of 3 - {[(4-Phenyl-2-thiazolyl) thio] methyl} -7- (2- (2-thienyl) acetamido] -3-cepherno-4-carboxylic acid in acetonitrile

7- [2- (2-Thienyl) acetamido] cephalosporanic acid (2.0 g, 5 mmol) and 4-phenyl-2-thiazolethiol (1.44 g, 7.5 mmol) in 35 mL of acetonitrile for 16 h reflux and protect from air moisture with a drying tube containing anhydrous calcium sulfate ("Drierite"). TLC indicates a complete transformation to a product with a single spot.

The reaction mixture was cooled to room temperature and stirred for 2 hours. The crystallized product is filtered off, washed with acetonitrile and dried. 2.25 g (85.6% yield) of product are obtained as a solid; mp 180 ° C (burping). The product was identified by NMR spectroscopy.

Example 59

3 - {[(5-Methyl-l, 3,4-thiadiazol-2-yl) thio] methyl} -7- [2- (lH-tetfazől-l-yl) -acetamide] -3-cephem Preparation of 4-cafanoic acid in nitrite

7- (2- (1H-Tetrazol-1-yl) -acetamidol-cephalosporanic acid <1.92 g, 5 rmillol) and 5-methyl-1,3,4-thiadiazol-2-thiol (0.797 g, 6 ril mole) (25 ml) was added to 25 ml of 15 ml of nitroethane in alumina and the mixture was heated in an oil bath to 95 ° C and stirred for 4 volumes. According to TLC, the product contains only trace amounts of cephalosporanic acid starting material (less than 2%). The reaction mixture was cooled to room temperature, filtered, washed with nitromethane and dried in vacuo. 2.11 g (92.5% yield) of product are obtained; mp 183.5 ° C (dec.). The product was identified by NMR spectroscopy.

Example 60

3 - {[(5-Methyl-l, 3,4-thiadiazol-2-yl) thio] methyl} -7- [2- (lH-tétraáol-l-yl) acetamido] -3-cephem Preparation of 4-carboxylic acid in propionitrile

7- (2- (1H-Tetrazol-1-yl) -acetamido] -cephalosporanic acid (1.92 g; 5 rtmol) and 5-methyl-1,3,4-thiadiazol-2-thiol (0.99 g) 7.5 ml) was added to 25 ml of propionitrile in 35 ml of alumina and heated to reflux (97 ° C) for 9 hours with stirring. Trituration and drying in vacuo gave 40 2.04 g (89.5% yield) of product;

186.5 ° C (dec.). The product was identified by NMR spectroscopy.

Example 61'- {[(1-Methyl-1 H -tetrazol-5-yl) thio] methyl} -7- (2-phenyloxy-2-phenylacetamido) -3-cephem-4 Preparation of carboxylic acid in 1,2-dichloroethane

7-Aminocephalosporanic acid (7-ACA) (13.62 g, 0.05 mol) was suspended in 100 ml 1,2-dichloroethane and trimethylsilylacetamide (26, z5 g, 0.2 mol) was added. . The reaction mixture was heated to 40 ° C and after the solid 55 dissolved, a cloudy solution was obtained which was cooled to 250 ° C.

2-formyloxy-2-phenylacetyl chloride (10.92 g, 0.055 mol) dissolved in 1,2-dichloroethane (10.92 g, 0.055 mol) was added dropwise over 20 minutes while the temperature was raised to 30 ° C. The reaction mixture was stirred for 2 hours at 100 mL

1.2-Dichloroethane was added and washed with water (3 x 100 mL). The aqueous washes were combined and 2 x 50 mL

1.2-dichloroethane. The dichloroethane phases are washed with 40 ml of water, combined, mixed with 2.0 g of activated carbon ("Darco G-60") for 20 minutes and treated with diatomaceous earth.

-17179967

Example 64 filtered through earth ("Hyflo"). The total volume of the solution containing the desired 7- (2-formyloxy-2-phenylacetamido) -cephalosporanic acid product is 375 ml.

The solution, consisting of about 250 ml of 1,2-dichloroethane and 22 g of 7- (2-formyloxy-2-phenylacetamido) -cephalosporanic acid, is concentrated to 336 g. 1-Methyl-1 H -tetrazol-5-thiol (6.39 g, 55 mmol) dissolved in 250 ml of 1,2-dichloroethane was added and the reaction mixture was heated to reflux. The water, presumably left over from previous washes, begins to drip back into the flask, forming an azeotrope in the refrigerator; a water trap is used to trap the water. TLC after 12 hours indicates near normal course of the reaction. The reaction mixture is then cooled to room temperature and seeded with 3 - {[(1-methyl-1H-15-tetrazol-5-yl) thio] methyl} -7 - [(2-formyloxy) -2-phenyl] for crystallization. -acetamido] -3-cephem-4-carboxylic acid product. After 2 hours, the product is filtered off and washed with 63 ml of 1,2-dichloroethane to give 13.90 g of product (56.7% yield based on 7-ACA). The product had a uniform TLC of 20 spots. The product was identified by its NMR spectrum, which was identical to that obtained in Example 16.

Example 62

Preparation of 3 - {[(1-Methyl-1H-tetrazol-5-yl) thio] methyl} -7- (2-formyloxy-2-phenylacetamido) -3-cephem-4-carboxylic acid 1, In 2-dichloroethane 30

7- (2-Formyloxy-2-phenylacetamido) -cephalosporanic acid (2.33 g, 5 mmol) and 1-methyl-1H-tetrazole-5-thiol (0.64 g;

5.5 mmol) is added to 25 ml of 1,2-dichloroethane and the mixture is refluxed for 12 hours. The reaction mixture was cooled to room temperature, then heated to reflux again and 10 mL of solvent was distilled off. Carbon tetrachloride (10 ml) was then added dropwise at reflux temperature (77 ° C). The mixture was allowed to cool to room temperature and stirred for 1 hour. The product is isolated by filtration,

Wash with 40% carbon tetrachloride in 1,2-dichloroethane (14 mL) and dry under vacuum at 50 ° C.

2.12 g of solid are obtained (86.5% yield). The NMR spectrum was identical to that obtained in Example 16.

Example 63

3 - {[(3-Benzyloxycarbonylaminomethyl) -1,2,4-triazol-50-5-yl) thio] methyl} -7- [2- (2-thienyl) - acetamido] -3-cephem-4-carboxylic acid in nitromethane

7- [2- (2-Thienyl) acetamido] cephalosporanic acid (2 g, 5 mmol) and 3- (benzyloxycarbonylaminomethyl) -1,2,4-55-triazole-5-thiol (2 g; 7.6 mmol) in 35 ml of nitromethane was heated at 80-90 ° C for 6 hours with stirring. The reaction mixture was cooled and the product was isolated by filtration. It was recrystallized twice from aqueous acetone.

1.2 g of cream-colored crystals are obtained; 174-60

178 ° C (dec.) (40% yield). Analysis for C ₂₅ H ₂₄ N ₆ O ₆ S ₃ :

C H N calculated 49.99 4.03 13.99 Found 50.20 4.03 13.76 18

Preparation of 3 - {[(1-carboxymethyl) -1H-tetrazol-5-yl) thio] methyl} -7- [2- (2-thienyl) acetamido] -3-cephem-4-carboxylic acid in acetonitrile

7- [2- (2-Thienyl) acetamido] cephalosporanic acid (1.0 g;

2.5 mmol) and 1- (carboxymethyl) -1H-tetrazol-5-thiol (0.61 g, 38 mmol) in 75 mL of acetonitrile was heated to reflux and 35 mL of solvent was distilled off. The reaction mixture was refluxed for 13 hours, cooled, filtered and concentrated under reduced pressure. The residue was dissolved in ethyl acetate, washed with 1 N hydrochloric acid and brine, and dried over sodium sulfate. Addition of hexane gives a precipitate which is separated and triturated with ether. 0.475 g of a tan solid were obtained (38% yield). The product is identified by comparing it with a certified sample obtained by other synthetic means.

Example 65

Preparation of 3 - {[(5-Methyl-1,3,4-thiadiazol-2-yl) thio] methyl} -7- (3-chloropropionamido) -3-cephem-4-carboxylic acid in nitromethane

7- (3-Chloropropionamido) cephalosporanic acid (1.0 g;

2.5 mmol) and 5-methyl-1,3,4-thiadiazol-2-thiol (0.4 g, 3 mmol) in 20 ml of nitromethane were immersed in an oil bath at 95-96 ° C. According to TLC, the reaction was complete in 3 hours. The reaction mixture was cooled to room temperature, filtered and concentrated in vacuo. A light red oil was obtained which crystallized at room temperature in 2 hours. The reaction mixture was triturated with ethyl acetate (15 mL), filtered, washed with ethyl acetate and dried. 0.64 g (55.2% yield) of product is obtained which is identified by IR, UV spectroscopy, titration, microanalysis and NMR spectroscopy.

Analysis for C ₁₆ H ₁₉ ClN ₄ O ₄ S ₃ :

C H N S cl Theoretical 41.51 4.14 12.10 20.78 7.66 Found 41.70 4.23 11.84 20.51 7.88

Example 66

- Preparation of {[(1-Benzyl-1H, 2,3-triazol-5-yl) thio] methyl} -7- [2- (2-thienyl) acetamido] -3-cephem-4-carboxylic acid 1,2-dichloroethane

7- [2- (2-Thienyl) acetamido] cephalosporanic acid (200 mg; 0.5 mmol) and 1-benzyl-1H-1,2,3-triazol-5-thiol (140 mg; 0.7 mmol) ) In 15 ml of 1,2-dichloroethane was heated at 65-70 ° C for 21 hours. The solvent was evaporated under reduced pressure, 25 ml of saturated sodium bicarbonate solution were added to the residue and extracted twice with ethyl acetate. To the remaining aqueous solution was added fresh ethyl acetate, cooled in an ice bath, and the pH was adjusted to 2.5 with 20% hydrochloric acid. The acidic product was extracted twice with ethyl acetate, and the combined ethyl acetate extracts were washed with brine and dried over anhydrous magnesium sulfate. The magnesium sulfate was filtered off and the ethyl acetate was evaporated under reduced pressure to yield 76 mg of ka-18179967. Compared to the authentic sample of the product and the starting material, TLC shows that the material consists of the product and starting material.

67-72. example

Replacement of 7-Methoxy-7- [2- (1H-tetrazol-1-yl) -acetamido] -cephalosporanic acid with Different Thiols

7-Methoxy-7- [2- (1H-tetrazol-1-yl) -acetamido] -cephalosporanic acid is reacted with various thiols in nitromethane. The procedures described above are used except that the products are purified from the unreacted starting material by high performance liquid chromatography. The products and their identification are as follows:

3 - {[(l-methyl-lH-tetrazol-5-yl) thio] methyl} -7-methoxy-7- [2- (lH-tetrazol-l-yl) acetamido] -3-cephem 4-carboxylic acid; UV maximum (ethanol) 273 Γημ (ε 9,082);

3 - {[(5-methyl-l, 3,4-thiadiazol-2-yl) thio] methyl} -7-methoxy-7- [2- (lH-tetrazol-l-yl) acetamido] - 3-Cephem-4-carboxylic acid, UV maximum (ethanol) 273 ιημ (ε 12,785).

Analysis for _C15 H ₁₆ N ₈ O ₅ S ₃ :

C H N calculated 37.18 3.33 23.13 Found 36.99 3.31 22.86

3 - {[(5-methyl-1,3,4-oxadiazol-2-yl) thio] methyl} -7-methoxy-7- [2- (1H-tetrazol-1-yl) acetamide] - 3-Cephem-4-carboxylic acid, UV maximum (ethanol) 269 mu. (ε 8,910). Analysis for C i jH ₁₆ N _g O ₆ S ₂ :

C

38.46

38.58

H

3.44

3.69

N

23.92

23.91

Calculated Found

3 - {[(l-carboxymethyl) -lH-tetrazol-5-yl) thio] methyl} -7-methoxy-7- [2- (lH-tetrazol-l-yl) acetamido] -3 -cefem-4-carboxylic acid, UV maximum (ethanol) 269 ηιμ (ε 7669). Analysis for C ₁₅ H ₁₆ N ₁₀ O ₇ S ₂ :

C H N calculated 35.16 3.15 27.33 Found 35.42 3.38 27.39

3 - {[(4,5-Dihydro-6-hydroxy-4-methyl-5-oxo-1,2,4-triazin-3-yl) thio] methyl} -7-methoxy-7- [2 - (1H-Tetrazol-1-yl) -acetamido] -3-cephem-4-carboxylic acid, UV 274 ιημ (s 11,967). Analysis for C ₁₆ H ₁₇ N ₉ O ₇ S ₂ :

maximum (ethanol)

C H N calculated 37,08 3.33 27.80 Found 36.98 3.43 27.79

Example 73

3 - {[(1-Methyl-1H-tetrazol-5-yl) thio] methyl} -7- [2- (4-ethyl-2,3-dioxo-1-piperazinylcarbonylamino) -2 -phenylacetamido] -3-cephem-4-carboxylic acid in nitromethane

7- [2- (4-Ethyl-2,3-dioxo-piperazinylcarbonylamino) -2-phenylacetamido] cephalosporanic acid hydrate (0.3 g, 0.5 mmol) and 1-methyl-1 H- Tetrazole-5-thiol (0.0725 g, 0.625 mmol) was added to 6 ml of nitromethane dried over alumina and the mixture was heated at 85 ° C for 12 hours under nitrogen. Nitromethane was evaporated and the residual brown foam was redissolved in sodium bicarbonate solution and washed twice with ethyl acetate. After addition of fresh ethyl acetate, the mixture was cooled to 0 ° C and adjusted to pH 2.3. The layers were separated and the aqueous layer was washed with ethyl acetate. The ethyl acetate layers were combined, washed with saturated sodium chloride solution, dried over anhydrous magnesium sulfate, filtered and evaporated. 130 mg (40% yield) of a pale yellow powder are obtained. The product was identified by its NMR spectrum:

81.2 (t, 3, -NCH ₂ CH ₃ , J = 7 Hz),

3.65 (m, 6, N-C / f ₂ CH ₃ and piperazinyl H),

4.0 (s, 3, tetrazole -

4.4 (s, 2, 3-Cf / ₂ S),

5.1 (d, 1, C ₆ -H, J = 6 Hz),

5.8 (d, 1, C ₇ -H, J = 8 Hz),

6.0 (d, 1, 34, J = 6 Hz),

7.4 (m, 5, phenyl-H),

8.4 (d, 1, -CHCON / 7-, J = 8 Hz) and

10.0 (d, 1, 35, J = 6 Hz).

Example 74

3 - {[(l-carboxymethyl) -lH-tetrazol-5-yl / thio] methyl} -7- [2- (4-ethyl-2,3-dioxo-piperazinyl-carbonylamino ) Preparation of -2-phenylacetamido] -3-cephem-4-carboxylic acid in 1,2-dichloroethane

7- [2- (4-Ethyl-2,3-dioxo-piperazinylcarbonylamino) -2-phenylacetamido] cephalosporanic acid hydrate (0.3 g, 0.5 mmol) was dissolved in alumina, previously dehydrated. 45 ml of 1,2-dichloroethane and the solution was heated to 95 ° C to remove water, 40 ml of solvent were distilled off with nitromethane (10 ml) and 1- (carboxymethyl) -1H-tetrazole-5. -thiol (0.160 g, 1 mmol) was added and the mixture was heated at 90 ° C for 12 hours under nitrogen. The reaction mixture was filtered, the solvent was evaporated, and ethyl acetate and a saturated sodium bicarbonate solution were added to the resulting gum. The mixture was washed twice with ethyl acetate, fresh ethyl acetate was added, cooled to 0 ° C and adjusted to pH 2.3 with 20% hydrochloric acid. The layers were separated and the aqueous layer was washed with ethyl acetate.

The ethyl acetate layers were combined and washed with a saturated sodium chloride solution, dried over anhydrous magnesium sulfate, filtered, concentrated and dried at room temperature. The product (123 mg) was identified by NMR spectroscopy:

1.2 (t, 3, -NCH ₂ CH ₃ , J = 7 Hz),

3.6 (m, 4, N-CH ₂ CH ₃ and piperazinyl 5-ff),

4.1 (m, 2, piperazinyl 4-H),

4.4 (s, 2, 3 — Cff ₂ S—),

5.1 (d, 1, C ₆ -H, J = 5 Hz),

5.3 (s, 2, tetrazole 1CZ7 ₂ COOH),

5.75 (d, 1, J = 6Hz),

5.9 (d, 1, 34),

7.4 (m, 5, phenyl H) and

9.9 (d, 1.36; J = 7 Hz).

Example 75

3 - {[(4,5-Dihydro-4-methyl-6-hydroxy-5-oxo-1,2,4-triazin-60-yl) thio] methyl} -7- [2- (4- Preparation of ethyl 2,3-dioxo-1-piperazinylcarbonylamino) -2-phenylacetamido] -3-cephem-4-carboxylic acid in nitromethane

7- [2- (4-Ethyl-2,3-dioxo-1-piperazinylcarbonylamino) -65-2-phenylacetamido] -ephephalosporanic acid hydrate (0.3 g; 0.6 g);

-19179967 mmol) and 4,5-dihydro-4-methyl-6-hydroxy-5-oxo-1,2,4-thiazin-3-thiol (0.111 g, 0.625 mmol) were dissolved in nitrogen at room temperature. .pitro-p) §tanban- é $, a. The batch roll is heated at 85 ° C for about 1 hour. The brown gum is poured out. The solvent is slowly evaporated to a yellow precipitate. Δ cooled for the rest of the time; filtered, washed with diethyl ether. The off-white solid product precipitates in two portions, which are identical by layer chromatography. All extensions ?. 80 mg (24%). The product was identified by NMR. - .1 δ 1.1 (t, 3, -NCH ₂ CH ₃ , J = 6 Hz),

3.3 (s, 3, triazine-C H ₃ ),.

3.65 (m, 4, N-C77 ₂ CH ₃ and piperazinyl 5-H), 4.0 (m, 2, piperazinyl 6-H),.

4.6 (s, 2, 3-CH ₂ S-),> \

5.1 (d, Ι, Ο, -τ-Η, J ~ 5Hz),

5.75 (m, .2,, 34 ^ ship with _beta-7 and C-csaport -Η), i

7.45 (m, 5, phenyl H),

9.5 (d, 1, -CHCONtt-, J = 8 Hz) and

10.0 (d, 1, 36).

76 · Example:

t'o'.í, '... . .....

3 - {[(5-Methylthio-1,3,4-thiadiazol-2-yl) thio] methyl} -7- [2- (1H-tetrazol-1-yl) acetamidoyl-3 cephem-4-carbonoS0V.QÍttO.-inétáll.tÍaÍl'.i:. 'u,, .'l · ..,. N, 7 ', 7', 7 '- [2r (1H-Tetrazol-1-yl) -acetamido] -cephalosporanic acid (15.0 g; 3S; 2, jnmol) and 5 - (methyl-). tip) -1., 3,4-thiadjazo-2i · -. >. λ Λ H>>'1, ·. ·, · _; ; ..:, i; . >.;>> 4, 3.

• -thiol (6.43 g, 39.2 mmol) was suspended in 500 ml of nitromethane previously passed through neutral alumina. The mixture was heated to 5 ° C. kept at this temperature for 6 hours, then cooled overnight, filtered, washed with 250 ml of nitromethane and dried at 45 ° C for 4 hours. 18.08 g (94.9%) of product are obtained.

This product was dissolved in 150 mL water and 4.0 mL acetic acid. PH in In sodium hydroxide solution (1-7 ml)

Adjust to 6.3 and turn the mixture to 1 for 1 hour. 257 ml of 10% 60% sodium lactate and 65 ml of ethanol are added and the mixture is allowed to stand for 45 minutes. It is stirred for 45 minutes, filtered, washed with 3x ethanol and dried.

The resulting product (18.1 g) was resuspended in 300 ml of ethanol for 3 hours, filtered, and washed with gtanol.

After drying, 1.4,85 g of product are obtained, which is identified by high pressure liquid chromatography (HPLC) ... , ..........,. ....... II. . , ..

77-84. example

3 - {[(5-Methyl-1,3,4-thiadiazol-2-yl) -thio] -methyl} -7- [2- (1H-tetrazol-1-yl) -acetamido] -3-cephem- 4 karbqnsav. , ...

in various solvents. j i,

7- [2- (1H-Tetrazol-1-yl) -acetamido] -cephalosporanic acid is reacted with 5-methyl-1,3,4-thiadiazole-2-thiol in various solvents. The essential details of the reaction are summarized in the following 30 (other reaction conditions are the same as in the previous examples). . ...

The amount of cephalosporanic acid Thioi. how much ¹ » ^; Solvent '' Ά. Temperature Time Yield, ° / o Analysis 1.91 g, i, · 1: 0.86 g Acetonitrile (molecule., Sieved). reflux 24hrs 87 190 ° C J>, 90 g - · 0.363 g nitXQrfeppzoj ,. 85 ° C and 100 ° C 16h 83 181 ° C (dec.) ' ^! i '1' ·. . H.i- ', ·· · 7 -í - 1 -. y; - ~ 4 · ^! 1 · t. 72h THE; THE, 0.96 g .. 03 g ... 1,2-dichloroethane and nitromethane 1,:, 1. 85 ° C 14hrs 88 182-183 ° C  rn; ' ' »Í E.g _t (decomposition), 0.96 g -0.5 g nitroethane 90 ° C 16h 84 177 ° C (dec.) £ 3.82, · i 1.98 g propylene carbonate 95 ° C 8h 6? 1.91 g 0.99 g of ethylene. _: 95 ° C 6hrs 22 2.02g * l, 0g epic acid. , _t .... _ti 85 ° C 5hrs 55 - 1.92 g 0.79 g. _52.5% and 47.5% nitromethane: a mixture of t-jjinitrq Ptop 95 ° C 3h 50 ' 89 -

* as the sodium salt

Example 85

Preparation of lithium 3 - {[(1-methyl-1H-tetrazol-5-yl) thio] methyl} -7- (2-hydroxy-2-phenylacetamido) -3-cephem-4-carboxylate glacial acetic acid,.

7- (2-Hydroxy-2-phenylacetamido) -cephalosporanic acid (0.48 g, 1 mmol) and 1-methyl-1H-tetrazol-5-yl (0.5 g;

(4.3 mmol) in 15 mL of glacial acetic acid at 84-86 ° C for 8 hours. The reaction mixture was cooled to room temperature and iodine (0.4 g, 1.6 mmol) was added to convert unreacted thiol to disulfide. After stirring for one minute at room temperature, the mixture was poured into a mixture of 100 ml of ethyl acetate and water and the excess iodine was removed by addition of sodium sulfite. The layers were separated and the ethyl acetate layer was washed with water (2.times.100 ml) and 20% sodium chloride solution (60.times.50 ml), then dried over anhydrous sodium sulfate. The ethyl acetate residue was removed on a rotary evaporator and the residue Dissolve in 10 ml of methanol. Ltipm acetate acetate dihydrate (0.21 g;

millimoles) was added and the mixture was stirred for 20 minutes at room temperature. The crystallized product

-20179967, washed with 5 ml of methanol and dried. White crystals (0.34 g, 69.4%) are obtained. The product was identified by its NMR spectrum:

δ 3.50 (ABq, J = 17 Hz),

3.92 (s, tetrazole -CH _} ),

4.20 (s, 3, - CH ₂ S-);

5.04 (d, C ₆ -H, J = 5 Hz),

5.24 (s, 0CWCONH -),

5.64 (d, _C7-H, J = 5 Hz), and

7.44 (s, 0- / 7).

The NMR spectrum is identical to the authentic sample prepared by reaction in aqueous medium.

Example 86

3 - {[(l-methyl-lH-tetrazol-5-yl) thio] methyl} -7- {2- [o- (tert-butoxycarbonyl-aminomethyl) phenyl] acetamido} - Preparation of 3-cephem-4-carboxylic acid in nitromethane

7- {2- [o- (tert-Butoxycarbonylaminomethyl) phenyl] acetamido} cephalosporanic acid (0.2 g, 0.386 mmol) and 1-methyl-1H-tetrazole-5-thiol (0.047 g). (0.405 mmol) in 5 ml of nitromethane at 85 ° C under nitrogen for 12 hours 3 - {[(1-Methyl-1H-tetrazol-5-yl) thio] methyl} -7- {2- [o- (tert-butoxycarbonylaminomethyl) phenyl] acetamido} -3-cephem-4-carboxylic acid is obtained. The product was identified by NMR spectroscopy:

δ 1.45 (s, 9, -C (C / 7 ₃ ) ₃ ,

3.65 (s, 4, 2-CH ₂ and - C // ₂ CONH),

3.85 (s, 3, tetrazole -CE7 ₃ ),

4.3 (m, 4, 3-CH ₂ S- and -C / 7 ₂ NHCOO-C, H ₉ ),

5.0 (d, 2, C ₆ -H, J = 4 Hz),

5.7 (q, I, C ₇ -H),

7.25 (s, 5, phenyl-H and -CON / -) and

8.9 (s, 1, -COO / 7).

Example 87

Preparation of 3 - {[(1-Methyl-1H-tetrazol-5-yl) thio] methyl} -7- (2-ureido-2-phenylacetamido) -3-cephem-4-carboxylic acid in nitromethane

7- (2-Ureido-2-phenylacetamido) -cephalosporanic acid (0.224 g, 0.5 mmol) and 1-methyl-1H-tetrazol-5-thiol (0.061 g, 0.525 mmol) in 8 mL of nitromethane at 85 ° C. Reaction at C for 12 hours 3 - {[(1-methyl-1H-tetrazol-5-yl) thio] methyl} -7- (2-ureido-2-phenylacetamido) -3-cephem-4 Carboxylic acid is obtained. The product was identified by NMR spectroscopy:

δ 3.5 (d, 2, - CH ₂ , J = 3 Hz),

3.83 (s, 3, tetrazole -CH ₃ ),

4.17 (d, 2, 3-CH ₂ S-, 1 = 3 Hz),

4.9 (d, 1, C ₆ -H, J = 5 Hz),

5.4 (d, 1, - CaffeCONH -, J = 8 Hz),

5.6 (m, 3, C ₇ -H and -Nff ₂ ),

6.7 (d, 1, 37, J = 8 Hz),

7.25 (m, 5, phenyl H) and

9.2 (d, 1.1 - CH2 COON -, J = 8 Hz).

Example 88

Preparation of 3 - {[(1-Methyl-1H-tetrazol-5-yl) thio] methyl} -7- (2-cyanoacetamido) -3-cephem-4-carboxylic acid in nitromethane

7- (2-Cyanoacetamido) -cephalosporanic acid (0.339 g, 1 mmol) and 1-methyl-1H-tetrazol-5-thiol (0.122 g, 1.05 mmol) in 10 mL of nitromethane at 85 ° C under nitrogen Reaction for 12 hours gives 3 - {[(1-methyl-1H-tetrazol-5-yl) thio] methyl} -7- (2-cyanoacetamido) -3-cephem-4-carboxylic acid. The product was identified by its NMR spectrum:

δ 3.5 (8.2.2-CH ^),

3.65 (s, 2, CNC / 7 ₂ CONH—),

3.95 (s, 3, tetrazole -CZ / ₃ ),

5.35 (s, 2, 3-CH ₂ S-),

5.1 (d, 1, C ₆ -H, J = 6 Hz),

5.75 (q, 1, (% -H, J = 4 Hz),

7.8 (s, 1, - COOH) and

8.7 (d, 1, -CH ₂ CON / 7-, J = 9 Hz).

Example 89

Preparation of 3 - {[(1-Methyl-1H-tetrazol-5-yl) thio] methyl) -7- [2- (m-chlorophenylthio) acetamido] -3-cephem-4-carboxylic acid nitromethane

7- [2- (m-Chloro-phenyl-thio) -acetamido] -cephalosporanic acid (0.390 g, 0.855 mmol) and 1-methyl-1H-tetrazole-5-thiol (0.104 g, 0.9 mmol) in 10 mL of nitro in methane at 85 ° C under nitrogen for 12 hours, 3 - {[(1-methyl-1H-tetrazol-5-yl) thio] methyl} -7- [2- (m-chlorophenylthio) - acetamido] -3-cephem-4-carboxylic acid is obtained. The product was identified by its NMR spectrum:

δ 3.6 (s, 2, 2-CH ₂ ),

3.75 (s, 2, - CWjCONH—),

3.9 (s, 3, tetrazole -CH ₃ ),

4.4 (s, 2, 3c / 7 S _2),

5.0 (d, 1, C ₆ -H, J = 5 Hz),

5.8 (q, 1, C ₇ -H, J = 4Hz),

7.15 (m, 4, phenyl-H),

7.7 (d, 1, -CH ₂ CON77-, J = 8 Hz);

8.5 (s, l, COO / 7).

Example 90

Preparation of 3 - {[(1-Methyl-1H-tetrazol-5-yl) thio] methyl} -7- [2- (phenylthio) acetamido] -3-cephem-4-carboxylic acid in nitromethane

7- [2- (Phenylthio) acetamido] cephalosporanic acid 0.422 g; 1 mmol) and 1-methyl-1H-tetrazol-5-thiol (0.122 g, 1.05 mmol) in 10 mL of nitromethane at 85 ° C under a nitrogen atmosphere for 12 hours were reacted with 3 - {[(1-methyl-1H) -tetrazol-5-yl) thio] methyl} -7- [2- (phenylthio) acetamido] -3-cephem-4-carboxylic acid is obtained. The product was identified by its NMR spectrum:

δ 3.7 (s, 4, - CZ _{7 2} CONH - and 2-C // ₂ ),

3.9 (s, 3, tetrazole -CH2),

4.35 (s, 2, 3-CH ₂ S-), '

4.9 (d, 1, C ₆ -H, J 6 Hz),

5.8 (q, 1, C ₇ -H, J = 4 Hz),

-21179967

7.25 (s, 5, phenyl-fi),

7.7 (d, 1, -CH ₂ CONff-, J = 8 Hz);

9.4 (s, 1, - ZOOM / Z),

Example 91

Preparation of 3 - {[(1-Methyl-1H-tetrazol-5-yl) thio] methyl} -7- [2- (2-dimethyl) acetamido] -3-cephem-4-carboxylate in n-butyl formate

7- [2- (2-Thienyl) acetamido] cephalosporanic acid (3.96 g, 10 mmol) and 1-methyl-1H-tetrazin-5-thiol (1.28 g, yl mmol) in 98 mL of n-butyl in formate at 84-86 ° C for 48 hours 3- {1- (1-methyl-1H-tetrazol-5-yl) thio] methyl} -7- [2- (2-thienyl) acetephamido] -j-cephem-4-carboxylic acid is obtained; mp 1673-168 ° C, yield 79.9%. The NMR spectrum of this product is identical to that of Example 5.

Example 92

Preparation of 3 - {[(1-Methyl-1H-tetrazol-5-yl) thio] methyl} -7- [2- (2-thienyl) acetamido] -3-cephem-4-carboxylic acid in toluene

The reaction of Example 91 was repeated, but toluene (150 mL) was used as solvent and the reaction was carried out at 84-86 ° C (136). The yield of the product with a melting point of 163-1633.5 is 88.5%. The NMR spectrum was identical to that of Example 5.

Example 93

Preparation of 3 - {[(1-Methyl-1H-tetrazol-5-yl) thio] methyl} -7- (tert-butoxycarbonylamino) -3-cephem-4-carboxylic acid band in nitromethane

7- (tert-Butoxycarbonylamino) -ephalosporanic acid (0.372 g, 1 mmol) and 1-methyl-10-tetrazol-5-thiol (0.122 g, 1.05 mmol) in 10 ml of nitromethane at 12 ° C for 12 hours. reacting to give 3 - {[(1-methyl-1H-tetrazol-5-yl) thio] methyl) -7- (tert-butoxycarbonylamino) -3-cetem-4-carboxylic acid. The product was identified by its NMR spectrum:

1.45 (s, 9, -C (Ctt ₃ ) ₃ ),

3.7. (s, 2, 2-CH3,

3.95 (s, 3, tetrazole -CH ₃ ),

4.4 (s, 2, 3— C # ₂ S—),

5.0 (s, 1, C ₆ -H), (5.6 (s, 2, Cy-H and -CH ₂ CON ₁ -) and (s, 1, · CO 2 H).

Example 94

3 - {[(l-methyl-lH-tetrazol-5-yl) thio] methyl} -7- {2- [4-ethyl-2,3-dioxo-piperazinyl-carbonylamino] -2- ( Preparation of p-hydroxyphenyl) acetamido} -3-cephem-4-carboxylic acid

7- [2- (4-Ethyl-2,3-dioxo-piperazinylcarbonylamino) -2- (p-hydroxyphenyl) -acetamide, p] -ephephosphoric acid (0.80 g, 0.306 mmol) and 1 -methyl-1 H -tetrazol-5-thiol (0.0374 g, 0.322 mmol) in 6 ml of nitromethane at 80 ° C for 12 hours was reacted with 3 - {[(1-methyl-1 H -tetrazol-5-yl) - thio] -methyl} -7- [2- (4-ethyl-2,3-dToxo-piperazinylcarbonylamino) -2- (p-hydroxy-phenyl) -acetamido] -3-cephem-4-carboxylic acid . The product is identified by its NMR spectrum:

δ 1.2 (t, 3, N-CH ₂ CH ₃ ),

3.6 (m, 6, 2-CH ₂ , piperazinyl 6-H and NC / 7 ₂ CH ₃ ). '^;'

3.9 (m, 2, piperazinyl 5-ff),

3.95 (s, 3, tetrazole -CH ₃ ),

4.4 (s, 2, 3-CH ₂ S-),

5.0 (d, 1, C _{6 -} H, J = 6 Hz), (<i, i, C ₇ - H, J = 6 Hz),

5.8 (d, 1, C //CÖNH, J = 8Hz),

6.8 (d, 2, phenyl-H, J = 8 Hz),

7.3 (d, 2, phenyl-H, J = 8 Hz),

8.3 (d, 1, CHCON / 7 -, J = 8 Hz) and

9.9 (d, 1, 35, J = 6 Hz).

Example 95

Preparation of 3 - {[(1,3,4-Thiadiazol-2-yl) thio] methyl} -7- [2- (1H-tetrazol-1-yl) acetarido] -3-cephem-4-carboxylic acid in Acetonitrile

7- (2- (1H-Tetrazol-1-yl) acetamido] cephalosporanic acid (0.38 g, 1.00 mmol) and 1'3,4-thiadiazol-2-thiol (0.15 g; 1 (27 mmol) in acetonitrile (15 mL) under nitrogen at reflux for 24 hours 3 - {[(1,3,4-thiadiazol-2-yl) thio] methyl} -7- [2- (1H-tetrazole) -1-yl) -acetamido] -3-cephem-4-carboxylic acid is obtained, which is identified by its NMR spectrum:

δ 3.74 (m, 2, 2-CH ₂ -),

4.48 (q, 2, 3 — C77 ₂ S—, J _AB = 14),

5.16 (d, 1, C ₆ - H, J = 5 Hz),

5.42 (s, 2, tetrazolyl-Ctt ₃ ),

5.76 (q, 1, C ₇ H, J ₇ = 5, J ₇ j> m ⁼ 8),

8.87 (s, 1, tetrazole ring),

9.40 (s, 1, thiadiazolthiol starting material),

9.53 (d, 1, - CH 2 CON 3 -, J = 8 Hz) and

9.57 (s, 1, thiadiazole ring).

Example 96 ^r '

3 - {[(5-Methyl-l, 3,4-thiadiazol-2-yl) thio] methyl} -7- [5-carbomethoxy-5- (2,4-dichlorobenzamido) -vaIeramido] - Preparation of 3-cephem-4-carboxylic acid in 1,2-dichloroethane

7- [5-Carbomethoxy-5- (2,4-dichlorobenzamide-p) -valeramido] -cellalosporanic acid (0.3 g; δ, 5 milliliters) and 5-methyl-1,3,4-thiadiazolyl-thiol (Ό (Ι g; 75 75 mmol) in 1,2-dichloroethane refluxed for ₁ h for ₂ h 3- (t-5-methyl-1,4,4-thiadiazol-2-yl) thio] methyl } -7- [555-carbomethoxy-5- (2,4-dichlorobenzamido) -valeramido] -3-cephem-4-carboxylic acid is obtained.

Example 97

Lithium 3 - {[(1-methyl-1H-tetrazol-5-yl) thio] methyl} -7- [2- (1H-tetrazol-1-yl) acetamido] -3-cephem- Preparation of 4-kafboxylate in acetonitrile

7- [2- (1H-Tetrazpl-1-yl) -acetamido] -cephalosporanic acid 65 (1.91 g, 5 mmol) and 1-methyl-1-t-tetrazol-5-thiol (0.7 g;

-2217996 'mmol) in acetonitrile (50 mL) was refluxed for 24 hours and cooled to room temperature. The solvent was removed on a rotary evaporator until approximately 10 mL of solution remained. After adding ethanol (40 mL), 0.3 g of lithium hydroxide in 10 mL of methanol was added. Lithium 3 - {[(1-methyl-1H-tetrazol-5-yl) thio] methyl} -7- [2- (1H-tetrazol-1-yl) acetamido] -3-cephem- The 4-carboxylate product crystallizes. The reaction mixture was stirred at room temperature for 45 minutes, the product was collected by filtration, washed with ethanol and dried at 40 ° C in vacuo to give 1.60 g (72% yield) of product which was identified by NMR.

Example 98

Preparation of 3 - {[(5-Methyl-1,3,4-thiadiazol-2-yl) thio] methyl} -7- (2-formyloxyphenylacetamido) -3-cephem-4-carboxylic acid 1,2-dichloroethane

7- (2-Formyloxy-2-phenylacetamido) -cephalosporanic acid (4.6 g; 8.8 mmol) and 5-methyl-1,3,4-thiadiazole-2-thiol (1.52 g, 11.5 mmol) in 1,2-dichloroethane (50 ml) was refluxed for 12 hours and then cooled to room temperature. 3 - {[(5-methyl-1,3,4-thiadiazol-2-yl) thio] methyl} -7- (2-formyloxy-2-phenylacetamido) -3-cephem-4 -Carboxylic acid product precipitates in the form of a dense paste. The mixture was diluted with 50 ml of 1,2-dichloroethane and stirred at room temperature for 4 hours. The product is isolated by filtration, washed with 1,2-dichloroethane until the filtrate is clear and dried in vacuo at 40-45 ° C. 2.8 g (63%) of product were identified by NMR.

Example 99

3 - {[(4,5-Dihydro-6-hydroxy-4-methyl-5-oxo-l, 2,4-triazin-3-yl) thio] methyl} -7- [2- (lH- Preparation of tetrazol-1-yl) -acetamido] -3-cephem-4-carboxylic acid in acetonitrile

7- (2- (1H-Tetrazol-1-yl) -acetamido] -cephalosporanic acid (8.9 g, 23.2 mmol) and 4,5-dihydro-6-hydroxy-4-methyl-5-oxo -1,2,4-Triazin-3-thiol (4.1 g, 25.8 mmol) in acetonitrile (200 mL) was refluxed for 23 hours, cooled to room temperature, and the product precipitated during the reaction was filtered, washed with acetonitrile (50 mL) and Dried at -50 ° C to give 9.70 g (86.6% yield) of product which was identified by its NMR spectrum.

Claims (15)

A process for the preparation of 7-acylamido-3-thiomethyl-3-cephem-4-carboxylic acids of formula I, wherein R ^{13 is} amidino, benzoyl, phenyl, C 1 -C 4 alkyl, or 4; R ^{1 is} hydrogen or methoxy; R ² is a group of formula 6; wherein -R ³ is -CH ₂ - (C 1-3 chloroalkyl), C 1-4 cyanoalkyl; The benzoyl group optionally substituted by halogen atoms of formula 7 is a protected 4-amino-4-carboxybutyl group ^: - where ''. * Represents an amino group protected as above; A group of Formula 8 wherein a and ¹ are independently hydrogen or hydroxy and m is 0 or 1; tetrazolyl-methyl; A group of Formula 9, wherein P is thienyl or a phenyl group of a compound of Formula 10, and a * is as defined above, and Q is a protected amino group of tert-butoxycarbonyl group, or an acylated amino group of formula wherein T is 13; R ^{13 in} R ⁴ represents a unit of R ⁶ with which the R 22 is optionally substituted with 5 or 6 membered 2 nitrogen and 1 oxygen or sulfur, or 3 or 4 nitrogen atoms, carbon atom -hepheroaromatic ring: or represents a unit with which the group 22 forms a 2-benzothiazolyl group, wherein the substituent is C 1-4 alkyl, benzyl, phenyl, OXO, hydroxy, a benzyloxycarbonylaminomethylcafboxymethyl or N-methylacetamido group, a compound of formula II wherein R ¹ and R ² are as defined above and R 1 is C 3 -C 3 alkyl, C 4 -C 6 cycloalkyl, amino, rriönovagy di (Cl-C3) alkylamino group is, formula 25 or formula 26 formula 24 group - and the formula R ¹³ -S-R ¹⁴ III a sulfur nucleophilic reagent - wherein R ¹³ is as defined above and R ^{14 is} hydrogen or a thiourea reaction, wherein the reaction is carried out in an organic solvent under anhydrous conditions. (Priority: March 7, 1977) - - =; The method of tooth opening according to claim 1, which is 3 - {[(1,3,4-thiadiazol-2-yl) thio] methyl} -7- (2- (1H-tetrazol-1-yl) - acetamido] -3-cephem-4-carboxylic acid by reaction of 7- [2- (1H-tetrazol-1-yl) acetamido] cephalosporanic acid with 1,3,4-thiadiazol-2-thiol, characterized in that reaction is carried out under organic solvent conditions in anhydrous conditions (Priority 7 March 1977). A process for the preparation of a compound according to claim 1 or 2 for the preparation of a compound of formula I wherein R ^{1 is} hydrogen and R ^{2 is a} 2-hydroxy-2-phenylacetamido group of a compound of formula II wherein R ¹ and R ² is a reaction of a compound of formula III and III, wherein the reaction is carried out in an organic solvent under anhydrous conditions. (Priority: March 7, 1977) The process of claim 3, which is 3 - {[(1-methyl-1H-tetrazol-5-yl) thio] methyl} -7- (2-hydroxy-2-phenylacetamido) -3- cephem-4-carboxylic acid by reaction of 7- (2-hydroxy-2-phenylacetamido) -ephalosporanic acid with 1-methyl-1H-tetrazole-5-thiol, characterized in that the reaction is carried out in an organic solvent under anhydrous conditions. (Priority: March 7, 1977) 5. The process of claim 1, wherein the temperature is from 50 ° C to 140 ° C. (Priority: March 7, 1977) The method of claim 5, wherein -23179967, characterized in that a temperature of 70-120 ° C is used. (Priority: March 7, 1977) 7. A process according to any one of claims 1 to 6, wherein the solvent is a hydrocarbon, alcohol, amide, ether, ketone, carboxylic acid, ester of carboxylic acid, halogenated hydrocarbon, nitro compound, nitrile or thioether. (Priority: March 7, 1977). 8. The process of claim 7, wherein the solvent is acetonitrile, 1,2-dichloroethane, methylene chloride, propionitrile, nitromethane, nitroethane, acetic acid, isopropyl acetate, butyl acetate, methyl isobutyl ketone, fluorobenzene, thiophene, methyl ethyl ketone, 1,1,2-trichloroethane, chloroform, benzene, carbon tetrachloride, isopropanol, nitrobenzene, propylene carbonate, ethylene carbonate, 2-nitropropane or n-butyl formate is used. (Priority: March 7, 1977) 9. Figures 1-8. A process for the preparation of a compound of the formula I as claimed in any one of claims 1 to 3 by reacting a compound of the formula II wherein R is methyl or amino and a compound of the formula III, wherein the reaction is carried out in an organic solvent under anhydrous conditions. (Priority: March 7, 1977) 10. Figures 1-9. A process for the preparation of a compound of formula I according to any one of claims 1 to 3, wherein R ^{1 is} hydrogen and R ^{2 is} 2- (2-thienyl) acetamido, a compound of formula II wherein R ¹ and R ² are as defined above, and III. by reaction of a compound of formula (I), characterized in that the reaction is carried out in an organic solvent under anhydrous conditions. 11. Figures 1-9. A process for the preparation of a compound of formula I according to any one of claims 1 to 3, wherein R ^{1 is} hydrogen and R ^{2 is} 2- (1H-tetrazol-1-yl) -acetamido, a compound of formula II wherein R ¹ and R ² are by reaction of the above compound and the compound of formula III, wherein the reaction is carried out in an organic solvent under anhydrous conditions. (Priority: March 7, 1977) 1.2. The process according to claim 11 is carried out on 3 - {[(5-methyl-1,3,4-thiadiazol-2-yl) thio] methyl} -7- [2- (1H-tetrazol-1-yl) ) -acetamido] -3-cephem-4-carboxylic acid 7- (2- (1H-tetrazol-1-yl) acetamido] -cephalosporanic acid and 5-methyl-1,3,4-thiadiazole-2-thiol characterized in that the reaction is carried out in an organic solvent under anhydrous conditions (Priority: March 7, 1977). 13 13. The method described in FIGS. A process for the preparation of a compound of formula I according to any one of claims 1 to 3, wherein R ^{1 is} hydrogen and R ^{2 is a} 2-formyloxy-2-phenylacetamido group, a compound of formula II - wherein R ¹ and R ² are as defined above. - a reaction of a compound of formula 10 and III, wherein the reaction is carried out in an organic solvent under anhydrous conditions. (Priority: March 7, 1977) The method of claim 13, which is an embodiment 15 Methods for Preparation of 3 - {[(1-Methyl-1 H -tetrazol-5-yl) thio] methyl} -7- (2-formyloxy-2-phenylacetamido) -3-cephem-4-carboxylic acid , 7- (2-formyloxy-2-phenylacetamido) -3-cephem-4-carboxylic acid and 1-methyl-1H-tetrazole-5-thiol, characterized in that the reaction is carried out in an organic solvent under anhydrous conditions. between. (Priority: March 7, 1977) 15. A process for the preparation of a compound of formula I according to any one of claims 1 to 3, wherein R ^{1 is} methoxy and R ^{2 is} 2-phenyl. 25-acetamido group by reaction of a compound of Formula II, wherein R ¹ and R ² are as defined above, with a compound of Formula III, wherein the reaction is carried out in an organic solvent under anhydrous conditions. (Priority: March 7, 1977) 30 16. Figures 1-9. A process for the preparation of a compound of formula I according to any one of claims 1 to 3, wherein R ^{1 is} hydrogen and R ^{2 is} 2-phenoxyacetamido, a compound of formula Π, wherein R ¹ and R ² are as defined above, and a compound of formula III. wherein the reaction is carried out in an organic solvent under anhydrous conditions. (Priority: March 7, 1977) 17. A method according to any one of claims 1 to 6 A process for the preparation of 40 compounds of Formula I wherein R * is hydrogen and R ^{2 is} 5-amino-5-carboxyvaleramido group, a compound of Formula II wherein R ¹ and R ² are as defined above and Formula III compound Characterized in that the reaction is carried out in an organic solvent under anhydrous conditions. (Priority: December 27, 1977)