CH539077A - Antibiotic cephalosporin cpds. - Google Patents

Abstract

Cehalosporin derivatives of the general formula (I): where R = hydrogen, or a group of formula (II), (III), (IV), (V), or (VI). In (II), X = protected amino, hydroxy, C1-C3 alkoxy, -COOH, or C1-C3 alkanoyl group. In formula (III), X = sulphur, oxygen, or a chemical bond, y = 0-2, z = 1-3. In (II), (III) and (IV), Q = hydrogen, C1-C3 alkyl, or alkoxy, cyano, nitro, hydroxy, chlorine, - bromine, fluorine, trifluoromethyl, -amino-(C1-C3)-alkyl, carboxymethyl (HOOC-CH2-) or carboxamidomethyl (H2NC(O)CH2-). In (V), X = oxygen or sulphur, and Z = hydrogen or together form another benzene ring. In figure (VI), R1 and R2 = hydrogen, methyl, 2- sydnone-3-(C1-C3)-alkanoyl group, (C1-C10)alkanoyl group, (C3-C10) alkenoyl group, (C1-C7)-alkyl-X-(C1-C3)-alkanoyl group (in which X = O or S), (C3-C7)-alkenyl-X-(C1-C3)-alkanoyl group (where X = O or S), (C2-C10)-halogenalkanoyl group (where the halogen is fluorine, chlorine or bromine), or a C2-C10 cyanoalkyl group, also In (I) R' = cyano group or the group OY where Y = C1-C10 alkyl, C3-C10 alkenyl, C3-C10 alkynyl, C5-C7 cycloalkyl, (C5-C7)-cycloalkyl-(C1-C3)-alkyl, C2-C10 haloalkyl, or a (C3-C10)-haloalkenyl or haloalkynyl group in which the halogen is bromine, chlorine, fluorine or iodine, a (C1-C3)-alkyl-X-(C1-C6)-alkyl (where X = O or S), (C2-C4)-alkanoyloxy (C1-C6)-alkyl, (C2-C4)-alkanoyl-(C1-C6)-alkyl, 2'-furyl (C1-C3)-alkyl, (2'-thienyl)-(C1-C3)-alkyl, 3'-tetrahydrofuryl, tetrahydrofuryl-(C1-C3)-alkyl, or by a phenyl, benzyl or phenylethyl group which may be substituted by either a (C1-C3)-alkyl, a (C1-C3)-alkoxy, (C1-C3)-alkoxycarbonyl, a fluorine, chlorine, bromine, nitro, cyano, or trifluoromethyl group. R" = hydrogen, a zwitterion charge, a salt with a pharmaceutically acceptable cation, a C4-C6 tert. alkyl group, a C5-C7 tert. alkenyl group, A C5-C7 tert. alkynyl group, a benzyl, benzhydryl, phthalimidomethyl, methoxybenzyl, nitrobenzyl, succinimidomethyl, trimethylsilyl or a phenacyl ester group. Antibiotics.

Description

  

  
 



   The present invention relates to a process for the preparation of t3-cephem derivatives, which are valuable as intermediates in a chemical process for the preparation of antibiotically active t3-cephalosporanic acid derivatives or which exert antibiotic effects as such.



   The semisynthetic production of 7-acylamidodeacetoxycephalosporin antibiotics from starting materials penicillin derivatives has recently become important because a process for converting penicillin sulfoxide esters into desacetoxycephalosporanic acid esters has been disclosed in US Pat. No. 3,275,626. Cephalosporin compounds, which are obtained from penicillins by this process, correspond to the following general formula:
EMI1.1
 where R is the remainder of an acylamido group in the 7-position and Rt is a hydrogen atom, a salt-forming cation, an ester group or a negative charge when the group COO- forms a salt with a cation either inside or outside the molecule.



   In an effort to expand and improve the properties and possible uses of these semi-synthetic cephalosporin substances derived from penicillin, attempts have been made to change the 3-methyl group of the above-mentioned t3-deacetoxycephalosporins into a group which of the resulting cephalosporin compound has a higher antibiotic effect against one or confers several gram-positive or gram-negative microorganisms.



  However, it has not hitherto been possible to convert a t3-deacetoxycephalosporin directly into a t3-cephalosporin, which contains a methyl radical carrying a functional group in the 3-position, in significant yield.



  There is therefore a need for another method for the production of more effective t3-cephalosporin antibiotics containing such a methyl group in the 3-position, which up to now could only be produced from cephalosporin C obtainable by fermentation and from the 7-aminocephalosporanic acid (7-ACA) obtained therefrom .



   The aim of the present invention is to create a key method for avoiding the difficulty encountered in converting t3-deacetoxycephalosporins into n3-cephalosporanic acid antibiotics which contain a methyl group carrying a functional group in the 3-position.



   The process according to the invention consists in that an A2-cephemdenvate of the formula
EMI1.2
 in which R is hydrogen, the group
EMI1.3
 wherein X denotes the amino or a protected amino group, the hydroxyl group, a (Cl-C3) alkoxy group, the carboxyl group or a (Cl-C3) alkanoyloxy group, the group
EMI1.4
 where X is oxygen, sulfur or a single bond, y is 0 or an integer from 1 to 2 and z is an integer from 1 to 3, or the group
EMI1.5
 and Q in each of the above groups is hydrogen, an alkyl or alkoxy group with 1 to 3 carbon atoms, the cyano, nitro, hydroxy or trifluoromethyl group, a chlorine, bromine or fluorine atom,

   an α-amino (Ct-Ca) alkyl group, the carboxymethyl or carbamoylmethyl group, or the group
EMI1.6
 where Y is sulfur or oxygen and each of the symbols Z is hydrogen or both Z together with the carbon atoms to which they are attached form a benzene ring, or the group
EMI1.7
 wherein each of R1 and R2 is hydrogen or the methyl group, or a sydnon-3-carbonyl, 2- (sydnon3) acetyl or 2- (sydnon-3) propionyl group, a (C1-Cio) -alkanoyl group, a (C-Cto) alkenoyl group, a (Cs-C7) alkyl-X- (C1-C3) alkanoyl group, where X is oxygen or sulfur,

   a (C3-C7) Alkenvl-X- (Ct to C3) alkanoyl group, where X is oxygen or sulfur, a (C2Clo) -haloalkanoyl group, where the halogen is a fluorine, chlorine or bromine atom, or a (C-Cto ) Cyanoalkanoyl group means, R 'the cyano group, the group OY, where Y is a (Ci-Cio) - alkyl group, a (C3-Ct0) alkenyl group, a (C3-C10) - alkynyl group, a (CsC7) cycloalkyl group, a (Cs up to C7) cycloalkyl (CtC3) alkyl group, a (C2-Cio) haloalkyl group,

   a (C-Cio) haloalkenyl group or a (C3-Cio) haloalkynyl group, where the halogen is a fluorine, chlorine or bromine atom, a (Ct-C3) alkyl-X- (C-Cs) alkyl group, where X is oxygen or Sulfur means a (C2-C) alkanoyloxy (C2-C6) alkyl group, a (C2-C4) alkanoyl (Ci-C6) alkyl group, a furyl (C1 to C3) alkyl group, a thienyl (C1- Cs) alkyl group, the 3 tetrahydrofuryl group, a tetrahydrofuryl (Ci-C3) alkyl group, the phenyl,

   Benzyl or phenylethyl group or one on the phenyl ring by a (C1-Cs) -alkyl group, (C1 to C3) alkoxy group, (Ci-C3) alkoxycarbonyl group, the fluorine, chlorine or bromine atom, the nitro, cyano or trifluoromethyl group substituted Phenyl, benzene or phenylethyl group and R "denotes hydrogen, one equivalent of a pharmaceutically acceptable cation, a (C4-Co) tert-alkyl group, a (Cs-C7) tert-alkenyl group, a (CsC7) tert-alkynyl group , denotes a benzyl, methoxybenzyl, nitrobenzyl, benzhydryl, phthalimidomethyl, succinimidomethyl, trimethylsilyl or phenacyl group,

   or an inner salt of a compound of the formula II having a free carboxyl group is converted into the corresponding X3 sulfoxide with an oxidizing agent and this is then reduced, with a corresponding cephem derivative of the formula
EMI2.1
 is obtained.



   The process according to the invention also comprises the preparation of internal salts of compounds of the formula I with a free carboxyl group and of acid addition salts of compounds of the formula I.



   The starting product of the formula II can be obtained by adding a t2-cephem-4-carboxylic acid ester of the formula
EMI2.2
 where R is an amino protecting group and R1 is the residue of an alcohol R1OH, in an essentially anhydrous organic solvent medium at a temperature in the range from about 0 C to the reflux temperature, preferably in the range from about 400 C to about

   1000 C, reacted with N-bromosuccinimide in order to obtain the corresponding 3-bromomethyl-t2-cephem-4-carboxylic acid ester as an intermediate, whereupon (b) the ester obtained as an intermediate is reacted with a nucleophilic substance to remove the bromine to displace the 3-bromomethyl intermediate product by the rest of the nucleophilic substance, a 2-cephalosporin ester product having a methyl group with a functional group in the 3-position being obtained.



  A nucleophilic substance is to be regarded as a substance which supplies a negatively charged group, or a neutral molecule which carries a lone pair of electrons and which enters into a nucleophilic substitution reaction with the intermediate 3-bromomethyl-t2-cephalosporin ester, namely below Formation of a 3- (nucleophilic methyl) -t2-cephalosporin ester. Countless examples of nucleophilic substances which can be used for this purpose have already been described in the cephalosporin antibiotic patent literature in connection with the formation of cephalosporin antibiotics which have the nucleophile on the methyl group in the 3-position.



   The starting product obtained (II) is then used in the process according to the invention, which (1) the oxidation of the t2 sulfide ester product by means of a peracid to a corresponding ns sulfoxy ester, (2) the reduction of the t3 sulfoxy ester with a reducing agent, such as. B.



  Sodium bisulfite or sodium dithionite, in the presence of an activator, such as. B. acetyl chloride, in an organic solvent such. B. acetic acid or dimethylformamide, to form the t3 sulfide ester.



   If desired, the antibiotic t3 sulfidic acid can be obtained by hydrolytic cleavage of the ester group of the t3 sulfide ester, e.g. B. by treatment with trifluoroacetic acid alone or with formic acid or acetic acid, whereby the effective cephalosporin antibiotic is obtained. If desired, mixtures of the antibiotically active 3-cephalosonic acid and the inactive 2-cephalosonic acid, as well as pharmaceutically acceptable salts thereof, in admixture for certain antibiotic purposes, e.g. B. for a topical antibiotic for open wounds in veterinary applications, in which one can dust the mixture on the wound, use.

  However, the materials can also be used in the form of ointments in order to inhibit the growth of various gram-positive or gram-negative microorganisms. On the other hand, the A2-cephalosporin ester can first be converted into a 3-ester and then this ester can be split into an A3-7-amino-3 nucleophilic-methyl) -cephalosporin ester, which in turn is N-acylated and becomes a t3-cephalosporin can be saponified.

 

  An example of such a compound which can be prepared by this working method is cephalothin, which is a much used, commercially available cephalosporin antibiotic and can currently only be obtained by fermentation from cephalosporin C and 7-ACA.



   The t2-deacetoxycephalosporin esters used in the preliminary stages can be obtained from various penicillin sources or cephalosporin sources by methods known per se. They can be obtained, for example, by treating corresponding 3-methyl-, 3-cephem-4-carboxylic acid esters as described in Example 4 of US Pat. No. 3,275,626 using bases.



  They can also be obtained by hydrogenating a cephalosporin ester which conducts itself from Cephalosporin C to give the corresponding t3-Desacetoxycep- halosporin ester, which is then treated with a base, such as. B. pyridine, in the cold, e.g. B. at 0 to 100 C, treated to isomerize the t3 double bond to a, 2-double bond. Attempts to brominate the allylic 3-methyl group of the t3-deacetoxycephalosporanic acid ester have so far failed. The present successful bromination of the t2-deacetoxycephalosporanic acid ester was therefore surprising and not to be foreseen.



   The amino protecting group represented by R in Formula III above may be any group known to protect the nitrogen atom to which it is attached against attack by the N -Bromosuccinimide. If the nitrogen is in the form of a free amino group, an excess of brominating agent is required to carry out the first work stage. The group R can e.g. B.



  a triphenylmethyl group (trityl group), a trimethylsilyl group or, best of all, an acyl group. Many of the acyl groups suitable for the present purposes are already known in the literature for the preparation of penicillin and cephalosporin antibiotics. Some such acyl groups can be subjected to bromination, a molar excess of N-bromosuccinimide being required in order to completely brominate the 3-methyl group in the 2-deacetoxycephalosporanic acid ester.

  If, for example, the thienylacetyl radical or the furylacetyl radical is used as the acyl protective group, the ring systems of these groups can be brominated in the first stage, such conditions not being disadvantageous for the present process, in particular if the 7-acyl group is split off in the further course of the process is said to form the corresponding 7-aminocephalosporin ester derivative. The preferred amino protecting group is an acyl group represented by the following formula
EMI3.1
 where m is an integer from 0 to 4, n is an integer from 1 to 4 and X is an oxygen or sulfur atom or where X is a chemical bond.

  Such acyl groups which have fluorine, chlorine, bromine or iodine atoms or alkyl radicals with 1 to 6 carbon atoms, alkoxy radicals with 1 or 2 carbon atoms, nitro groups, cyano radicals or trifluoromethyl groups as substituents on the phenyl carbon atoms are likewise preferred.



  The acyl group can also have methyl groups instead of one or two hydrogen atoms on the carbon atoms in the m or n groups. Some examples of such preferred acyl groups are: phenylacetyl, phenoxyacetyl, Phenylmercaptoacetyl, benzyloxyacetyl, 3 -Methylphenylbutyryl, 4-Propylb enzylmercaptoacetyl, B enzylmercaptopropionyl, phenylpropionyl, Phenyläthylmercaptopropionyl, Phenylbutoxybutyryl, 3-Fluorphenoxyacetyl, 4-bromophenylacetyl, 2-Chlorbenzyloxypropionyl, phenyl-a, a -dimethylacetyl, 4-nitrophenylmercaptoacetyl, 3-cyano-phenylpropionyl, 4-trifluorophenoxyacetyl, etc.



   Countless other compounds which form amino groups protecting acyl groups which can be used in the R position belong to the prior art. For example, such compounds are described in U.S. Patent Nos. 2,479,295 to 2,479,297 and 2,562,407 to 2,562,411, and 2,623,876.



   The radical R1 represents the radical of an ester-forming alcohol. The alcohol used to form these esters should be one which can be removed by methods known per se, e.g. B. by means of dilute aqueous base or by using trifluoroacetic acid or by hydrogenation in the presence of a palladium catalyst or rhodium catalyst on a suitable carrier, such as. B.



  Carbon, barium sulfate or aluminum oxide so that the cephalosporin is not broken down. The preferred ester groups are those mentioned above.



   In the first step in the production of the starting product, the esterified 2-deacetoxycephalosporin (III) used as the starting material is mixed with N-bromosuccinimide in an organic liquid medium and stirred or otherwise kept in motion until the 3-bromomethyl product is formed . The reaction is preferably carried out in the presence of azo-bis-isobutyronitrile as a catalyst or initiator to increase the yield, but the process can also be carried out without a catalyst. The reaction proceeds more quickly when the mixture is heated, which is why it is preferred to work in a temperature range of 400.degree. C. to 1000.degree. The temperature should be below the decomposition point of the starting material and product.

  Temperatures that are too low also cause the reaction to take place too slowly for effective operation. The N-bromosuccinimide is generally present in at least the stoichiometric amount based on the G2-deacetoxycephalosporanic acid ester, since the latter is more expensive. The amount of azobis-isobutyronitrile and other initiators that are used is not important. Organic liquid diluents for the reaction mixture are carbon tetrachloride or mixtures thereof with chloroform, tetrachloroethane, methylene chloride, benzene, toluene, xylene, heptane, etc.



   Examples of the new t2-3-bromomethyl compounds which arise as intermediate products include: 4-methoxybenzyl-7-phenoxyacetamido-3-bromomethyl-t2-cephem-4-carboxylate, 4-methoxybenzyl-7-phenylacetamido-3-bromomethyl -t, 2-cephem-4-carboxylate, benzhydryl-7-phenylmercaptoacetamido-3-bromomethyl-t, 2-cephem-4-carboxylate, tert.

   -Butyl-7- (4'-nitrophenoxyacetamido) -3 -bromomethyl-n2- cephem-4-carboxylate, 3,5 -dimethoxybenzyl-7- (3'-chlorophenylmercaptoacetamido) -
3-bromomethyl-t2-cephem-4-carboxylate, tert-butyl-7- (4 'iodophenoxyacetamido) -3-bromomethyl-t2-cephem-4-carboxylate, 3-methoxybenzyl-7- (3', 5'- dibromophenoxyacetamido) -3 -bromo- methyl-Q2-cephem-4-carboxylate, phthalimidomethyl-7-benzamido-3 -bromomethyl- / X2-cephem-
4-carboxylate, succinimidomethyl-7- (4'-trifluoromethylphenoxyacetamido) -
3-bromomethyl-2-cephem4-carboxylate, tert-butyl-7- (4'-fluorophenylmercaptopropionamido) -3-bromomethyl-t2-cephem-4-carboxylate, tert-butyl-7-phenoxyacetamido-3-bromomethyl- t2-cephem
4-carboxylate, p-nitrobenzyl-7-phenylacetamido-3-bromomethyl-t2-cephem-
4-carboxylate,

   1,1-Dimethyl-2-propenyl-7-phenoxyacetamido-3-bromo methyl-nz-cephem-4-carboxylate and 1,1-dimethyl-2-propynyl-7-phenoxyacetamido-3-bromo methyl-t2-cephem- 4-carboxylate.



   After the end of the bromination, the 2-3-bromomethylcephalosporin ester obtained as an intermediate can either be isolated from the reaction mixture or treated in the same reaction mixture for the second step with a nucleophilic reagent, the bromine in the 3-methyl position being replaced by the nucleophile. In the organic medium, the ti2-3-bromomethylcephalosporin ester obtained as an intermediate product is mixed or mixed with the nucleophile in question until the bromine atom is replaced by the nucleophile. With certain nucleophilic reagents, the mixture should be heated to accelerate the reaction.



  Heating the reaction mixture to reflux in a low boiling solvent, e.g. B. at less than 1000 C, for example in acetone, acetonitrile, dimethylformamide, dimethyl sulfoxide, benzene, heptane or the like, favors the conversion. Depending on the type of nucleophilic reagent used, the bromide may appear in a by-product or it may be present as a salt of a quaternary nitrogen in the cephalosporin compound. The products can be in the form of oils or crystalline, solid substances. The product can be purified by washing with nonsolvents or by chromatographic methods. On the other hand, the product (11) in the crude reaction mixture can be immediately treated with a suitable oxidizing agent, e.g.

  B. m-chloroperbenzoic acid, treat to form the t3-cephalosporin sulfoxydester therefrom, whereupon it is treated with a suitable reducing agent, such as. B. sodium bisulfite or sodium dithionite in a mixture with acetyl chloride and dimethylformamide, whereby the corresponding cephalosporin ester (I) is formed with a function in the position. Certain t3-cephalosporin esters, e.g. B. the acetoxymethylcephalothin esters, can be used as such for antibiotic therapy to combat bacterial infections. In most cases, however, it will be preferred to remove the protective ester group in a manner known per se to obtain ns-cephalosporanic acid or a zwitterion derivative. The obtained cephalosporanic acid or



  the zwitterion can be used as such or in pharmaceutically acceptable salts, e.g. B. in the alkali metal salts, such as the sodium or potassium salt, or in soluble or insoluble amine salts, can be converted, which depends on the type of treatment with the desired salt. Insoluble salts with 1,4-bis- (aminomethyl) -cyclohexane are valuable as depot antibiotics for the administration of cephalosporin, since in this way the cephalosporin antibiotics are made slowly accessible in the bloodstream. In such form, the cephalosporin is generally injected intramuscularly, in doses of 0.5 to 1 g. Water soluble
Salts of cephalosporin antibiotics, such as.

  B. the sodium,
Monoethanol or diethanolamine cephalosporin salts can be administered parenterally using total doses of about 1 to about 6 g of active cephalosporin antibiotic per day for a patient weighing about 70 kg.



   The nucleophilic reactants which can be used to replace the bromine from the 3-bromomethyl group can be any compounds or substances which provide a negatively charged group, or neutral molecules which carry a lone pair of electrons, such as pyridine, and which have a nucleophilic substitution with the 3-bromomethyl group. Many compounds containing nucleophilic groups are already known which are suitable for producing cephalosporin antibiotics which carry a nucleophilically substituted methyl group in the 3-position. This is evident in particular from the patent literature.

  Some nucleophilic reactants are more suitable than others, since the nucleophilic groups which the reactants provide produce an increased antibiotic effect of the resulting cephalosporin compound compared to the corresponding cephalosporin compounds which do not contain nucleophilic groups.



   Suitable nucleophilic reactants of the type defined above are those compounds which contain nucleophilic oxygen, sulfur, nitrogen or carbon atoms, as is illustrated by the following groups of nucleophilic reactants, which are preferred.



   1. Compounds of the formula:
EMI4.1
 wherein M denotes a hydrogen atom or an alkali metal, Rt denotes an alkyl group having 1 to 6 carbon atoms, an alkenyl group having 3 to 6 carbon atoms or a carbo-Ct- or -C2-alkoxy group and A denotes a hydrogen atom or a carboxyl group. Such compounds are e.g. B. 4-methyl-, 4-ethyl-, 4-isopropyl-, 4-n-propyl-, 4-isobutyl-, 4-n-butyl-, 4-n-amyl-, 4-neopentyl-, 4- n-Hexyl-, 4-allyl-, 4- (2-butenyl) -, 4-carbomethoxy- and 4-carboethoxy-piperazino-dithio-carboxylates and other compounds, which are described in detail in US Pat. No. 3,239,516 are.



   2. Pyridine-type compound of the following formula:
EMI4.2
 wherein Rs is a hydrogen, fluorine, chlorine or bromine atom, an alkyl radical having 1 to 3 carbon atoms, a carboxy radical, a carboxamide residue (-CONH2), an N-methylcarboxamide residue or an N, N-dimethylcarboxamide residue and R4 is a hydrogen atom, a fluorine atom , a chlorine atom or a bromine atom or an alkyl radical having 1 to 3 carbon atoms, such as. B. pyridine, a-picoline, ß-picoline, 3,5-dichloropyridine, 4-bromopyridine, 3-fluoropyridine, lutidine, picolinic acid, nicotinic acid, isonicotinic acid, nicotinamide, N-methylnicotinamide, N-methyl-isonicotinamide, N, N-dimethylnicotinamide and N, N-dimethylisonicotinamide, as e.g. B. in US Pat. No. 3,280,118.

 

   3. Thiourea, thioacetamide, thiosemicarbazide, N-AI- kylthioureas with 1 to 6 carbon atoms in the alkyl radical, N-phenylthiourea and alkali metal salts of such Ver compounds as they are, for. B. in US Pat. No. 3,278,531.



  4. Dithiocarbamates and xanthates of the formulas:
EMI5.1
 and
MS-CS-OR7 in which each of the radicals R5 and R6 is an alkyl radical with 1 to 6 carbon atoms or both radicals R5 and Re together with the nitrogen atom to which they are connected represent a ring with 4 or 5 methylene carbon atoms, while R7 is a primary one An alkyl group having 1 to 12 carbon atoms, a secondary alkyl group having 3 to 12 carbon atoms, a cycloalkyl group having 3 to 7 carbon atoms and M is an alkali metal, such as. B.

  Na trium N, N-dimethyldithiocarboxylate, sodium piperidino-dithiocarboxylate and other dithiocarboxylate reactants, as they have been known from US Pat. No. 3,258,461, also sodium ethoxydithiocarboxylate and other alkali metal xanthogenates, which according to the method of Drawert, Born, Ber. 93, 3064 (1960), as well as the alkali metal xanthate reagents with nucleophilic character.



   5. M-N3, M-SCN, M-NO2 or M-CN, where M is an alkali metal, such as e.g. B. sodium and potassium azide, thiocyanate, nitrite or cyanide, as disclosed for example in US Pat. No. 3,274,186, or where M is divalent copper, such as cuprocyanide, or a silver ion, such as. B. silver cyanide, which can also be used.



   6. Compounds of the formula
EMI5.2
 where each symbol X is an oxygen or sulfur atom and Rs is a hydrogen atom, an alkyl radical with 1 to 6 carbon atoms, an alkenyl radical with 2 to 6 carbon atoms, a phenyl radical, a benzyl radical or a phenylethyl radical or such phenyl radicals, benzyl radicals and phenethyl radicals which are attached to their ring carbon atoms Trifluoromethyl, nitro, cyano radicals, alkyl radicals with 1 to 3 carbon atoms, alkoxy radicals with 1 or 2 carbon atoms, fluorine atoms, chlorine atoms or bromine atoms, or where Rs denotes the following radical:
EMI5.3
 wherein Y) is NH, -0- or -S-, y is the number 0 or 1 and M is an alkali metal or an alkaline earth metal.

  Such compounds are, for example, alkali metal or alkaline earth metal alkanoate salts of formic acid, acetic acid, propionic acid, butyric acid, pentanoic acid, hexanoic acid, acrylic acid, methyl acrylic acid, crotonic acid, 3-hexenoic acids, and the thio analogs of such acids, such as. B. thioacetic acid, thiopropionic acid, dithiobutyric acid, dithioacetic acid, thiophenecarboxylic acid, 3-thiophenecarboxylic acid, 2-furancarboxylic acid, etc., also the alkali metal or alkaline earth metal salts, such as. B. the calcium and magnesium salts, benzoic acid, thiobenzoic acid, dithiobenzoic acid, phenylacetic acid, phenylthioacetic acid, phenyldithioacetic acid, phenylpropionic acid, phenylthiopropionic acid, phenyldithiopropionic acid, and substituted acids such.

  B. 4-trifluoromethylbenzoic acid, 3-nitrophenyl acetic acid, 2-cyanophenylthioacetic acid, 4-methylphenyldithioacetic acid, 3-ethylphenylpropionic acid, 4-methoxyphenyl acetic acid, 3-chlorophenylthioacetic acid, 2-fluorophenyldithio-pyro acetic acid, 2-fluorophenyldithio-pyro-acetic acid, 2-fluorophenyldithio-pyro-acetic acid, 2-fluorophenyldithio-pyro-acetic acid, 2-fluorophenyldithio-pyro-acetic acid, 2-fluorophenyldithio-pyro-acetic acid, 2-fluorophenyldithio-pyro-acetic acid, 2-dithio-3-bromopylacetic acid, 2-cyanophenylthioacetic acid, 4-methylphenylthioacetic acid -Pyrrolyldithioacetic acid, 2-thienyl acetic acid, 3-thlenyl acetic acid, 2-thienylthioacetic acid, 2-furyl acetic acid, 3-furyl acetic acid and 3-furyl thioacetic acid, as described, for example, in US Pat. No. 3,218,318 and 3,261,832.



   7. Amines selected from primary alkylamines with 1 to 6 carbon atoms in the alkyl group, dialkylamines with 1 to 6 carbon atoms in each alkyl group and trialkylamines with a total of 3 to 12 carbon atoms, monocyclic, cycloaliphatic amines with 5 to 7 carbon atoms, monocyclic, saturated , nitrogen-containing heterocyclic amines with 4 to 6 carbon atoms and a total of 5 to 7 ring-forming atoms, as well as aromatic amines of the formula:
EMI5.4
 wherein each of the symbols Rs and Rio denotes hydrogen or methyl and R11 denotes a hydrogen atom, an alkyl radical with 1 to 6 carbon atoms or an alkoxy radical with 1 or 2 carbon atoms, such as amines of the following type:

  Methylamine, ethylamine, isopropylamine, hexylamine, dimethylamine, diethylamine, dibutylamine, dihexylamine, piperidine, morpholine, 4-methylpiperazine, thiomorpholine, trimethylamine, triethylamine, tributylamine, clopentylamine, cyclohexylamine, hexine, piperidine, aniperazine, N-pyrrolamine N-methylaniline, N, N-dimethylaniline, 4-methylaniline, 4-methyl-N-methylaniline, 3-propylaniline, etc.



   8. Compounds of the formula
Rt2-X-H where X is oxygen or sulfur and R12 is the residue of an alcohol or mercaptan, which preferably has a molecular weight below about 200; R12 can, however, have the following meanings, for example: hydrogen, C1- to Cio-alkyl, C3- to Cie-alkenyl, Ci- to Cto-alkynyl, C2 to Cio-haloalkyl, C3- to Cie-haloalkenyl and Csb to Cie-haloalkynyl, where the halogen is fluorine, chlorine, bromine or iodine, C1- or Ci-cyanoalkyl, C5- to C7-cycloalkyl, C5- to C7-cycloalkyl-Ct- to Ce-alkyl, phenyl, benzyl,

   Phenylethyl, where the phenyl, benzyl and phenylethyl are substituted on ring carbon atoms by Ci- to Cs-alkyl, Ci- to Cs-alkoxy, C1- to Cs-alkyloxycarbonyl (i.e.



  -COO-alkyl with 1 to 3 carbon atoms in the alkyl group), fluorine, chlorine, bromine, nitro, cyano or trifluoromethyl can be substituted, Ci- to Cs-alkyl-X-C2- to -Ce-alkyl, where X is oxygen or Sulfur means Ci- to C4-alkanol-, Ci- to Ce-alkyl, C2- to C4-alkanoyloxy- Ci- to -Ce-alkyl, C1- to C6-alkyloxycarbonyl-Cr- to -Ce- alkyl,
EMI6.1
 where Y is -0-, -S- or> NH and y is 0 to 2, and the like, as well as the sodium, potassium and calcium salts of all these compounds have the pKa values of less than 11.



   Examples of such compounds are water, hydrogen sulfide, alcohols and mercaptans such as methanol, ethanol, isopropanol, tert-butanol, hexanol, octanol, decanol, allyl alcohol, 3-butenyl alcohol, 3-hexenyl alcohol, 4-octenyl alcohol, 9-decenyl alcohol, propargyl alcohol, 3-butynol, 3-hexynol, 4-octynol, 4-chloro-1-butanol, cyclohexanol, cyclopentanol, cycloheptanol, cyclopentylethanol, cyclohexylpropanol, 2-chloroethanol, 2-bromopropanol, 2-chloro-2-butenyl alcohol, 2-chloro-3 -butynyl alcohol, 2-pyrrolylethanol, 3-pyrrolylpropanol, furanol, 2-thienylmethanol, thiophen-ol, 2-thienylethanol, 2-furylmethanol and 3-furylethanol, 2-cyanoethanol, 3-cyanopropanol, benzyl alcohol, anisyl alcohol, p-nitrobenzyl ethanol -Trifluoromethylphenylethanol, p-cyanobenzyl alcohol, propyloxyethanol, methoxyethanol, ethoxyethanol,

   Propionylethanol, butanoyloxypropanol, butoxycarbonylethanol, phenol, p-toluene, 3-chlorophenol, 4-ethoxyphenol, resorcinol and the corresponding mercaptan analogues thereof; A few examples of such mercaptan analogs are methyl mercaptan, allyl mercaptan, phenyl mercaptan, benzyl mercaptan, furanethiol, thiophenethiol, 2-thienylethyl mercaptan, 3-thienylmethyl mercaptan, 2-furylethyl mercaptan and the alkali metal or alkaline earth metal salts, which have lower values than the phenols and alkaline earth metal salts of phenols and alcohols of such alcohols .



   9. N-Aminoalkyldithiocarbaminate compounds of the formula:
EMI6.2
 where z is 2 or 3, R19 is C1- to Ce-alkyl and Rt4 and R15 are each C1- to Co-alkyl or Rt4 and Rts together with the nitrogen atom to which they are bonded form a ring with 4 to 6 ring carbon atoms, where R14 and Rts together contain not more than 8 carbon atoms, and M is hydrogen or an alkali metal; Examples of these compounds are M-methyl-N (2'-dimethylaminoethyl) dithiocarbaminate, N-methyl-N- (2'-diethylaminoethyl) -dithio-carbaminate, N-hexyl-N- (2'-di-n-propylaminoethyl ) dithiocarbaminate, N-methyl-N- (2'-morpholinoethyl) -dithiocarbaminate, N-methyl-N- (2'-piperidinopropyl) -dithiocarbaminate, as they are, for.

  As described in U.S. Patent No. 3,239,515.



   Specific examples of starting products of the formula II and of nucleophilic reagents which can be used to produce the same by replacing the bromine from the 3-bromomethylcephalosporin intermediates are: 7- (4'-trifluoromethylphenoxyacetamido) -3 - (4-methylpiperazi nodithiocarboxylate) -t2 -cephem-4-carboxylic acid ester from 4
Methyl piperazinodithiocarboxylate;

   7- (2'-Phenoxyacetamido) -3 - (4 ', 4'-dimethylpiperazinodithiocarboxylate) -t2-cephem-4-carboxylic acid ester iodide from 4-methylpiperazinodithiocarboxylate with subsequent treatment with methyl iodide; 7- (3'-chlorophenoxyacetamido) -3- (pyridinomethyl) -A2-cep-hem-4-carboxylic acid ester bromide from pyridine; 7- [2 '- (3-chlorophenylmercaptoacetamido)] - 3- (3'-carbamylpyridinomethyl) -, 2-cephem-4-carboxylic acid ester bromide
Nicotinamide; 7- (4'-Phenylbutoxyacetamido) -3- (2'-pseudothioureidomethyl) - t2-cephem-4-carboxylic acid ester bromide from thiourea; 7- [4 "4-tert-butoxycarbamidomethyl) -benzamido] -3- (2'-imidazolylthiomethyl) -t2-cephem-4-carboxylic acid ester from 2
Mercaptoimidazole;

  ; 7- (Phenoxyacetamido) -3 - (azidomethyl) -2-cephem4-carboxylic acid ester from sodium azide and the product obtained by reducing the 3-azidomethyl group to the 3-aminomethyl group by catalytic hydrogenation with palladium or platinum as catalyst; 7- (phenylmercaptoacetamido) -3-aminomethyl-t2-cephem-4-carboxylate from sodium azide with subsequent reduction of the azide group to the amino group;

   7- (Phenylmercaptoacetamido) -3-N, N-dimethyldithlocarbamyl methyl-t2-cephem-4-carboxylic acid ester from N, N-dimethyl dithiocarbamate, sodium salt; 7- (ethylmercaptoacetamido) -3 -N, N-dimethyldithiocarbamyl-ethyl-2-cephem4-carboxylic acid ester from N, N-dimethyl dithiocarboxylate, sodium salts; 7- (3'-chlorophenoxyacetamido) -3- (picolinoylthiomethyl) -A2-cephem-4-carboxylic acid ester from sodium thiopicolinate; 7th <(4'-Nitrobenzyloxyacetamido) -3 - (4'-nitrobenzoylthiomethyl) -t2-cephem-4-carboxylic acid ester from sodium 4-nitro thiobenzoate; 7- (3'-Methoxyphenylacetamido) -3- (4'-tert-butylpiperazino-thiocarbonylthiomethyl) -t2-cephem-4-carboxylic acid ester from sodium 4'-tert-butylpiperazinodithlocarboxylate;

  ; 7- (3'-Chlorophenoxyacetamido) -3- (4'-methylpiperazinothlocar bonylthiomethyl) -ssi2-cephem-4-carboxylic acid ester from sodium 4-methylpiperazinodithiocarboxylate; 7- [5 '- (4 "-Cyanophenoxy) -pentanamido] -3 - (4-isopropylpiper azinothiocarbonyl-thiomethyl) -t2-cephem-4-carboxylic acid ester from sodium 4-isopropylpiperazinodithiocarboxylate; 7- (2'-fluorophenylacetamido ) -3- [N-sec.-butyl-N- (2'-dimethyl- aminoätltyD-aminothlocarbonylthiomethylj-2-cephem-4-carboxylic acid ester from sodium N-sec.-butyl-N- (2'-dimethylaminoethyl) -aminodithiocarboxylate; 7- (caprylamido) -3- [N-methyl-N-2- (N'-ethyl-Nn-hexylamino-ethyl) -aminothiocarbonylthiomethyl] -na-cephem4-carboxylic acid ester from sodium N-methyl N- [2- (N'-ethyl-Nn hexylamino) ethyl] aminodithiocarboxylate;

  ; 7- [2 '- (phenylisopropoxy) acetamido] -3- (acetoxymethyl) -A2-cephem-4-carboxylic acid ester from potassium acetate; 7- (Phenoxyacetamido) -3- (propionoxymethyl) -t2-cephem-4-carboxylic acid ester from sodium propionate, and 7- (naphthylmercaptoacetamido) -3- (hexanoyloxymethyl) -A2-cephem-4-carboxylate from potassium hexanoate, the
Ester groups are as defined above.

 

   According to the present process, for example, p-methoxybenzyl-7-phenoxyacetamido-3-cyanomethyl-t3cephem-4-carboxylate can be obtained by oxidizing p-methoxybenzyl-7-phenoxyacetamido-3-cyanomethyl-t2-cephem-4-carboxylate to the corresponding 1-oxide (sulfoxide), e.g. B. with m
Chloroperbenzoic acid in a suitable solvent, and subsequent reduction of the sulfoxide with sodium dithionite or equivalent reducing agents in the presence of acetyl chloride.



   The 7-amino-3-cyanomethyl- and -3-oxymethyl ether-t3-cephem-4-carboxylic acid esters, which are referred to in English as nuclei type compounds, can also be obtained by splitting off the 7-acyl group or another 7-amino blocking group, which was present during the above oxidation and reduction stages can be prepared. For example, p-nitrobenzyl-7-amino-3-ethoxymethyl-t3-cephe.m-4-carboxylate can be obtained by p-nitrobenzyl-7-phenoxyacetamido-3-ethoxymethyl-t3-cephe.m-4-carbo - Treated xylate in the presence of a tertiary amine with phosphorus pentachloride and then with an alkanol, such as methanol, and finally with water to bring about the cleavage.



   The 7-amino-3-cyanomethyl- and -3-oxymethyl ether-t3-cephem-4-carboxylic acids and the 7-acylamido-3-cyanomethyl and -3-oxymethyl ether-t5-cephem-4-carboxylic acids can be obtained by cleaving the ester groups from the corresponding esters are prepared by known methods or by means of the methods described here.



   Production of the starting products t2 acid production
3.63 g (0.01 mole) methyl-7- (phenoxyacetamido) -3-methyl.



     t5-cephem-4-carboxylate (prepared from Desacetoxyceph V and diazomethane) in 100 cm3 of a mixture of pyridine and water in the ratio 1: 1 were cooled in an ice water bath. One equivalent of 1N sodium hydroxide solution was added and the mixture was stirred in the cold for 5 hours. After dilution with 100 cm of water and 100 cm3 of ethyl acetate, the mixture was cooled and acidified to pH = 2.5 with 20% hydrochloric acid. The ethyl acetate was removed and the aqueous layer extracted once with ethyl acetate. The combined organic layers were cooled, water was added and the pH was adjusted to 8.2 with solid sodium bicarbonate.

  The aqueous layer was separated, washed once with ethyl acetate, then cooled, layered with ethyl acetate and acidified to pH 2.5. The ethyl acetate was separated and the aqueous layer washed with ethyl acetate and the combined organic layers washed twice with sodium chloride solution, dried over magnesium sulfate, filtered and evaporated to give a foam.



   The foam-like product was dissolved in ethyl acetate and crystallized therefrom, giving 1.6 g (45 O / o) 7- (phenoxy acetamido) -3-methyl-t2-cephem-4-carboxylic acid with a melting point of 180 to 1830 C (decomposition) .



   Esterification of t2-deacetoxycephalosporanic acid a) p-methoxybenzyl-3-methyl-7-phenoxyacetamido-3-cephem-4-carboxylate.



   A suspension of 1.75 g (0.005 mol) of A3-desacetoxyceph V (7-phenoxyacetamido-3-methyl-t2-cephem-4-carboxylic acid) and 700 mg (0.005 mol) of p-methoxybenzyl alcohol in 20 cm3 of methylene chloride a condensation agent solution was added with stirring, which consisted of 1.23 g (5% excess) of dimethylformamide-dineopentyl acetal (DMF-dineopentyl acetal) in methylene chloride. Complete dissolution occurred in a few minutes. The reaction mixture was stirred at room temperature overnight. The solvent was removed and benzene added. After heating to effect dissolution, the mixture was allowed to stand at room temperature.



   After a crystalline secondary reaction product had been separated off, the benzene mother liquor was diluted, washed three times with bicarbonate solution and twice with sodium chloride solution, dried over magnesium sulfate, filtered and evaporated. The residue was crystallized from carbon tetrachloride and gave 4-methoxybenzyl-7-phenoxyacetamido-3-methyl-, -cephem-4-carbo.xylate. First, 1.15 g of melting point 108 to 1120 ° C. and 0.16 g of melting point 107 to 1110 ° C. from the mother liquor were obtained; the yield was 55%.



   b) Introduction of functional groups in p-methoxy benzyl-3-meth-yl-7-phenoxyacetamido-t2-cephem-4-carboxylate.



   A mixture of 235 mg of p-methoxybenzyl-3-methyl-7-phenoxyacetamido-t2- and -t3-cephem-4-carboxylate, 90 mg of N-bromosuccinimide (NBS), 14 mg of azobisisobutyronitrile and 30 cm3 of carbon tetrachloride was added for 14 hours under nitrogen heated to reflux, protecting it from light. The reaction mixture was cooled and filtered and the golden yellow filtrate was evaporated to dryness. 25 mg of potassium acetate and 15 cm 3 of acetone were added to the residue, and this mixture was heated to reflux for 6 hours under nitrogen and protected from light and then stirred overnight at room temperature.

  The dark red-brown solution was evaporated to dryness, and the residue was taken up in chloroform, filtered and evaporated to give 250 mg of a red-brown oil.



   This oil was purified twice by preparative thin layer chromatography to give 103 mg of a mixture of the 2- and t3-p-methoxybenzyl esters of Ceph V.



   Cleavage of the mixed -L1 "ester product with trifluoroacetic acid in benzene gave a material containing Ceph V acid, a known antibiotic, as shown by thin layer chromatography and a bioautograph of a paper chromatogram.



   Example 1 a) The oxidation of the t2-t4 ester mixture with m-chloroperbenzoic acid in chloroform gave the corresponding p methoxybenzyl-3-2cetoxymethyl-7- (2'-phenoxyacetamido) - / 3-cephem-4-carboxylate-1-oxide of Melting point 161 to 163 C (from methanol) in high yield.



   b) reduction.



   A solution of 500 mg of the sulfoxide, namely 4-methoxybenzyl-3-acetoxymethyl-7- (2'-phenoxyacetamido) -t, 2-cephem4-carboxylate-1-oxide, in 40 cm3 of dimethylformamide (DMF) was 10 cm3 Acetyl chloride and then mixed with 3 g of sodium dithionite. An exothermic reaction resulted. After stirring for 4 hours at room temperature to ensure complete reaction, the mixture was cooled. diluted with benzene and mixed with aqueous sodium bicarbonate solution.

  After the vigorous evolution of gas had ceased, more benzene was added and the organic layer separated and washed well with aqueous sodium bicarbonate solution and then with sodium chloride solution, dried over magnesium sulfate, filtered and evaporated to leave 635 mg of a brown oil.

 

   This product was purified on a column of silica gel with 15% water content. 253 mg of an oil were obtained whose magnetic nuclear magnetic resonance, infrared and ultraviolet spectrum was the same as the corresponding spectra of p-methoxybenzyl-7- (2'-phenoxyacetamido) -t3-cephem-4-carboxylate, which was obtained by fermentation of 7-aminocephalosporanic acid (7-ACA). This oily ester product could be crystallized from ether and gave 195 mg of a material with a melting point of 118 to 119 ° C. A mixed melting point of this material with known samples of the same compound gave no melting point depression.



   Example 2
Cleavage of the p-methoxybenzyl ester to form Ceph V.



   A solution of 105 mg of p-methoxybenzyl-7- (phenoxy-acetamido) -3-acetoxymethyl-t3-cephem-4-carboxylic acid ester and 22 mg of anisole in 10 cm3 of dry benzene was mixed with 0.5 cm3 of trifluoroacetic acid. The mixture was stirred under nitrogen at room temperature for 2 hours and then evaporated to dryness. The residue was taken up in ethyl acetate and extracted three times with aqueous sodium bicarbonate solution.



   The ethyl acetate solution was evaporated to give 55 mg of a neutral product.



   The bicarbonate extracts were cooled, covered with ethyl acetate and brought to pH = 2.4 with 20% hydrochloric acid. The ethyl acetate layer was separated and the aqueous portion extracted once with ethyl acetate. The combined organic layers were washed twice with sodium chloride solution, dried, filtered and evaporated to give 86 mg of a colorless oil which, in a thin layer chromatogram, showed the same Rf value as 7- (phenoxyacetamido) -3 -acetoxymethyl-n3-cep- hem- 4-carboxylic acid. The chloroform solution of this oil separated a solid which was removed by filtration.



  The filtrate was evaporated, leaving 65 mg of product which was found to be 7- (phenoxyacetamido) -3-acetoxy methyl-, L3-cephem-4-carboxylic acid, since its nuclear magnetic resonance spectrum is identical to that of a known sample of this compound, a known one Anti biotilsums, which previously could only be made from the product obtained by fermentation of 7-ACA.



   Preparation of nP- and t3-methyl-3-acetoxymethyl-7-phenoxyacetamido-2-cephem-4-carboxylate mixture
91 mg (0.25 millimoles) of crystalline methyl 3-methyl-7-phe noxyacetamido-t2-cephem-4-carboxylate, 45 mg (0.25 millimoles) of N-bromosuccinimide, 7 mg of azobisisobutyronitrile and 15 cm3 of carbon tetrachloride were in a Brought 50cc round bottom flask to make methyl 3-bromomethyl-7-phenoxyacetamido-t2-cephem-4-carboxylate. The mixture was refluxed under nitrogen for 14 hours, protected from light, cooled, filtered and evaporated to dryness. 25 mg (0.25 millimoles) of potassium acetate and 15 cm3 of acetone were added to the residue.



  The mixture was refluxed under nitrogen for 6 hours, protected from light, and stirred at room temperature overnight. After the solution had been evaporated to dryness, the residue was taken up in chloroform and filtered. Evaporation of the solvent gave 130 mg of a golden colored oil.



   This oil was separated into its components by preparative thin layer chromatography, with 11 mg of starting material being recovered and 70 mg (yield 65%) of a mixture of methyl 3-acetoxymethyl-7-phenoxyacetamido-t2-cephem-4-carboxylate and Methyl 3-aceto xymethyl-7-phenoxyacetamido-ZS3-cephem-4-carboxylate was obtained; the structure was confirmed by nuclear magnetic resonance, and the ratio of 02 to t3 compound was 3: 1.



   Preparation of a mixture of A2- and t3-methyl-3-azidomethyl-7-phenoxyacetamido-ceph-2-em-4-carboxylate methyl 3-bromomethyl-7-pheno; acetamido-2-cephem-4-carboxylate was as in previous section herge. After evaporation of the carbon tetrachloride solvent, 16 mg of sodium azide (0.25 millimoles) and 15 cm3 of acetone were added to the residue. The mixture was refluxed for 6 hours and then stirred at room temperature overnight. The acetone was evaporated and the residue taken up in chloroform and filtered. Removal of the solvent gave 123 mg of a brown oil.



   The brown oily crude product was purified by preparative thin layer chromatography. There was thus obtained 79 mg of a material containing both the starting material and the azido product as shown by the infrared spectrum. The investigation by nuclear magnetic resonance and infrared spectrum showed that this material had 16/0 starting material (mixture of A2 and A3 compounds in a ratio of 1: 1) plus the desired mixture of methyl-3-azidomethyl-7-phenoxyacetamido-t2- cephem-4-carboxylate and methyl-3-azidomethyl-7-phenoxyacetamido t3-cephem-4-carboxylate (t2 to t3 compound in a ratio of 3: 1) in a yield of 65%.



   A solution of p-methoxybenzyl-7-phenoxyacetamido 3-bromomethyl-t2-cephem-4-carboxylic acid ester in dry benzene is mixed with a solution of 2 molar equivalents of anhydrous trimethylamine in dry benzene and stirred for 3 hours at room temperature. After this time, the benzene solvent is separated from the precipitated product by decantation. The precipitated residue is dissolved in a mixture of water and acetone in a volume ratio of 1: 1 and separated on an ion exchange resin column (Dowex 1 X 1 in the acetate form).

  The fraction flowing out first is freeze-dried and gives a glass-like product which is composed of a mixture of p-methoxybenzyl-7-phenoxyacetamido-3- (N, N, N-trimethylammoniummethyl) -, 2-cephem-4-carboxylic acid ester and its t3 - Isomers in the form of the acetate salts.



   According to the procedure of the above section, p-methoxybenzyl-7- (phenoxyacetamido) -3- (bromomethyl) -A2- cephem-4-carboxylate is converted with methylamine to p-methoxybenzyl-7 (phenoxyacetamido) -3- (N-methylaminomethyl) - , 2-cephem-4-carboxylic acid ester, mixed with the corresponding t3 ester, reacted.



   According to the procedure of the above section, the ester p-methoxybenzyl-7- (p-methylphenylacetamido) -3bromomethyl-2-cephem-4-carboxylate with 2 molar equivalents of semicarbazide of the formula I12N-NH-CO-NH2 is converted into p methoxybenzyl-7- (p-methylphenylacetamido) -3 - [(N'-carboxylic acid amido) -hydrazinomethyl] -t, 2-cephem-4-carboxylate reacted.



   This ester can be converted into a valuable antibiotic by treating it with dilute base to form a mixture which contains the antibiotic 7- (phenoxyacetamido) -3 - [(N'-carboxamido) -hy drazinomethyl] -t3-cephem -4-carboxylic acid ester, whereupon the ester group is removed by treating the ester mixture with trifluoroacetic acid.

 

   According to the procedure of the above section, the following t2-cephalosporic acid esters are prepared with the specified nucleophilic reagents: B enzyhydryl-7- (phenoxyacetamido) -3- (N-hydroxyaminomethyl) -ssS2-cephem-4-carboxylate from benzhydryl-7- ( phenoxyacetamido) -3-bromomethyl-2-cephem-4-carboxylate and
Hydroxylamine; p-Methoxybenzyl-7- (4'-methylphenoxyacetamido) -3- (hydrazino-methyl) -t2-cephem-4-carboxylate from p-methoxybenzyl-7- (4'-methylphenoxyacetamido) -3- (bromomethyl) -2 - cephem-4-carboxylate and hydrazine;

   p-Methoxybenzyl-7- (4-methoxyphenylmercaptoacetamido) -3 [N- (4-methylpiperazino) -1-aminomethyl] - A2-cephem-4-carboxylate from p-methoxybenzyl-7- (4-methoxyphenylmer captoacetamido) -3- (bromomethyl) -t2-cephem-4-carboxylate and 1-amino-4-methylpiperazine; Methoxybenzyl 7- (4'-nitrophenylacetamido) -3 - (N-morpholino methyl) -t2-cephem-4-carboxylate from bis- (3,5-dimethoxy-phenyl) -methyl-7- (4'-nitrophenylacetamido) - 3 -bromomethyl a2-cephem-4-carboxylate and morpholine; and tert-butyl 7- (4'-bromothiophene-2'-acetamido) -3 - (N-methylanilinomethyl) -t2-cephem-4-carboxylate from tert-butyl-7 (4'-bromothiophenacetamido) -3- (bromomethyl) -t2-cephem-
4-carboxylate and N-methyl aniline.



   Other nitrogen-containing nucleophilic reagents used in
Preparation of the t2 esters with functional groups in 3
Position that can be used are piperazine, imidazole,
Imidazoline, pyrazole and other compounds, such as nucleophilic oxygen, sulfur, nitrogen or carbon atoms, their use for the production of
Cephalosporin antibiotics is known from the literature.



   A solution of p-methoxybenzyl-7- (phenoxyacetamido) -3-bromomethyl-t, 2-cephem-4-carboxylate in dry benzene is mixed with a benzene solution of 2 molar equivalents
Methyl mercaptan mixed. The mixture is during 3
Stirred for hours at room temperature to allow the reaction to take place
End runs. The reaction mixture is evaporated in vacuo to remove the solvent. The residue contains the crude product p-methoxybenzyl-7- (phenoxy acetamido) -3- (methylthiomethyl) -t, 2-cephem-4-carboxylate.



   According to the procedure of the above section, the following 2-cephalosporin esters with functional groups in the 3-position are obtained with the specified nucleophilic sulfur reagents: 4-methoxybenzyl-7- (phenoxyacetamido) -3- (phenylthiomethyl) -t2-cephem -4-carboxylate from 4-methoxybenzyl-7 (phenoxyacetamido) -3-bromomethyl-t2-cephem-4-carboxylate and thiophenol;
Benzhydryl 7- (phenyl-a, a-dimethylacetamido) -3- (mercapto methyl) -A2-cephem-4-carboxylate from benzhydryl-7- (phenyl-a, a-a-amethylacetamido) -3- (bromomethyl) - n2-cephem-
4-carboxylate and hydrogen sulfide;

  ;
4-methoxybenzyl-7- (4'-nitrophenoxyacetamido) -3- (allylthio methyl) -t, 2-cephem-4-carboxylate from 4-methoxybenzyl 7- (4'-nitrophenoxyacetamido) 3-bromomethyl-t2-cephem-
4-carboxylate and allyl mercaptan; tert-Butyl-7- [p-tert-butoxycarbamido) -phenylmercaptoacet- amidol -3- (carbomethoxymethylthiomethyl) -t2-cephem-4-carboxylate from tert-butyl-7- [p- (tert-butoxycarbamido) phenylmercaptoacetamido] -3-bromomethyl-t2-cephem-4-carboxylate and methylthioglycolate;

  ; Benzyl 7- (3'-chlorophenoxyacetamido) -3 (N, N-dimethylaminoethylthiomethyl) -t2-cephem-4-carboxylate from benzyl-7 (3'-chlorophenoxyacetamido) -3-bromomethyl-t2-cephem-4-carboxylate and N , N-dimethylaminoethyl mercaptan.



   These t2-cephalosporin esters with functional groups in the 3-position can be converted into a mixture containing antibiotically active 3-cephalosporanic acid derivatives by treating the t2-cephalosporin esters with a dilute base, such as, for example, triethylamine, whereby a mixture which contains the t3-cephalosporin ester and the corresponding t2-cephalosporin ester, which mixture is then treated with trifluoroacetic acid in benzene to produce the mixture of the corresponding A2- and 3-cephalosporanic acids.



   A solution of p-methoxybenzyl-7- (phenoxyacetami do) -3-bromomethyl-t2-cephem-4-carboxylate in dry
Benzene is admixed with a thoroughly dried suspension which contains 1.1 molar equivalents of sodium N, N-dimethyldithiocarbaminate suspended in benzene, while stirring.



  The mixture is stirred for about 3 hours at room temperature. The insoluble solid material is removed by filtration and the benzene solution of the reaction product is concentrated to a small volume and chromatographed over silica. The eluate fractions, which contain p-methoxybenzyl-7-phenoxyacetamido-3-methyl-t2- and 3-cep- hem-4-carboxylic acid esters, are discarded, while the p-methoxybenzyl-7- (phenoxyacetamido) -3- (N , N-dimethylaminothiocarbonylthiomethyl) -t2-cephem-4-carboxylic acid ester-containing eluate fractions are collected and evaporated to dryness.

  The product obtained is treated with pyridine to form an equilibrium mixture of the p-methoxybenzyl-7-phenoxyacetamido-3-N, N-dimethylami- nothiocarbonylthiomethyl-a2- and -t3-cephem-4-carboxylic acid esters, and then the Ester mixture treated with trifluoroacetic acid in benzene.



   According to the procedure of the above section, the following t2-cephalosporin esters with a functional group in the 3-position are prepared with the specified nucleophilic reagents: 1,1-Dimethylpropinyl-7- (phenylmercaptoacetamido) -3- (N'-methyl-N-piperazinothiocarbonylthiomethyl) - / S2-cephem-
4-carboxylate from 1,1-dimethylpropynyl-7- (phenylmercap toacetamido) -3-bromomethyl-2-cephem-4-carboxylate and
N-methyl piperazinodithiocarboxylate; 4-Methoxybenzyl-7- (4'-nitrophenoxyacetamido) -3 - (thioaceto-xymethyl) -t2-cephem-4-carboxylate from 4-methoxybenzyl 7- (4'-nitrophenoxyacetamido) -3-bromomethyl-nr-cep- hem -4-carboxylate and sodium thioacetate;

  ; tert-Butyl 7- (octanoylamino) -3- (benzoylthiomethyl) -A2-cephem-4-carboxylate from tert-butyl-7- (octanoylamino) -3 bromomethyl-T2-cephem-4-carboxylate and potassium thio benzoate; 4-methoxybenzyl-7- (phenoxyacetamido) -3 - (picolinoylthiomethyl) -t2-cephem-4-carboxylate from 4-methoxybenzyl-7 (phenoxyacetamido) -3-bromomethyl-t2-cephem-4-carboxylate and sodium thiopicolinate; and benzhydryl-7- (phenyl-a, a-dimethylacetamido) -3- [N- (s-dimethylaminoethyl) -N-methylthiocarboxylthiomethyl] 2 cephem-4-carboxylate from benzhydryl-7- (phenyl-a, a- dimethyl acetamido) -3-bromomethyl-t2-cephem-4-carboxylate and sodium N- (β-dimethylaminoethyl) -N-methyldithio carboxylate.



   These t2-cephalosporin esters with a methyl group bearing a functional group in the 3-position can easily be converted into the corresponding tS3-cephalosporanic acid derivatives with an antibiotic effect by means of the procedure described above.



   A solution of p-methoxybenzyl-7- (phenoxyacetamido) 3-bromomethyl-t2-cephem-4-carboxylate in dry benzene is treated with a solution of 1.1 molar equivalents of thiourea in dry benzene. After stirring for 5 hours at room temperature so that the reaction is complete, two drops of tetramethylammonium hydroxide in methanol are added and the mixture is allowed to stand for a further hour, the product being a crude mixture of p-methoxybenzyl-7 - (phenoxyacetamido) -3 -formamidinothiomethyl-t2- and -t3-cephem-4-carboxylate hydrobromide is obtained.

 

   In accordance with the procedures of the above section, the specified products are obtained if the thiourea is replaced by the following nucleophilic reagents: 4-methoxybenzyl-7- (phenoxyacetamido) -3 - (2'-pyrirnidylthio-methyl) -, 2-cephem-4 carboxylate from 2-mercaptopyrimide in; 4-methoxybenzyl-7- (phenoxyacetamido) -3 - (2'-thiazolylthio-methyl) -n2-cephem-4-carboxylate from 2-mercaptothlazole; 4-methoxybenzyl-7- (phenoxyacetamido) -3- (2'-benzimidazolyl-thiomethyl) -t2-cephem-4-carboxylate from 2-mercapto benzimidazole;

  ; 4-methoxybenzyl-7- (phenoxyacetamido) -3- (2'-pyridylthiomethyl) -, 2-cephem-4-carboxylate from 2-mercaptopyridine; 4-methoxybenzyl-7- (phenoxyacetamido) 3 - (2'-benzothiazolyl-thiomethyl) -t2-cephem-4-carboxylate from 2-mercaptobenz thiazole; 4-methoxybenzyl-7- (phenoxyacetamido) -3- (2'-imidazolylthio-methyl) -, 2-cephem-4-carboxylate from 2-mercaptoimidazole; 4-Methoxybenzyl-7- (phenoxyacetamido) -3- [3 '- (1', 3 ', 4'-thiadiazolyl)] - thiomethyl-t2-cephem-4-carboxylate from 2-Mer capto-1,3 , 4-thiadiazole;

  ; 4-methoxybenzyl-7- (phenoxyacetamido) -3- [2 '- (1', 3 ', 4'-oxadiazolyD] 4hlomethyl-2-cephem-4-carboxylate from 2-mer capto-1,3,4 -oxadiazole; and 4-methoxybenzyl-7- (phenoxyacetamido) -3 - (2'-pyrazinylthio-methyl) -a2-cephem-4-carboxylate from 2-mercaptopyrazine.



   These n2-cephalosporin esters are suitable as intermediates for the production of the corresponding t3-sulfide acid antibiotics by means of the above-described equilibrium mixture formation / ester cleavage process.



   A solution of 4-methoxybenzyl-7- (phenoxyacet amido) -3-bromomethyl-n2-cephem-4-carboxyl in dry acetonitrile is mixed with 1.1 molar equivalents of dry sodium nitrite and the mixture is stirred for 30 hours at room temperature, with 4 Methoxybenzyl-7 (phenoxyacetamido) -3- (nftromethyD-2-cephem-4-carboxylate is formed. If this t2-cephalosporin ester is converted into the corresponding t3-cephalosporanic acid derivative, the latter shows antibiotic action against S. lutea on eioautography The nuclear magnetic resonance spectrum of this product shows a 3'-methylene group.



   By following the procedure of the above paragraph, replacing the sodium nitrite with the indicated nucleophilic alkali metal, copper or silver salts, the following A2-cephalosporin-4-esters are obtained: 4-methoxybenzyl-7- (phenoxyacetamido) -3 - ( cyanothiomethyl) - na-cephem-4-carboxylate from sodium thiocyanate; 4-methoxybenzyl-7- (phenoxyacetamido) -3- (cyanomethyl) -A2-cephem-4-carboxylate from cuprocyanide in dimethyl sulfoxide; and 4-methoxybenzyl-7- (phenoxyacetamido) -3 - (cyanomethyl) -t2-cephem-4-carboxylate from silver cyanide.



   These t2-cephalosporin esters can be used as starting products for the formation of the corresponding 3-cephalosporic acid antibiotics or the salts of such acids according to the procedures described in Example 1.



   A mixture of 4-methoxybenzyl-7-phenoxyacetamido 3-bromomethyl-t2-cephem-4-carboxylate in dry benzene and 1.1 molar equivalents of pyridine is mixed at room temperature for 5 hours, with 4-methoxybenzyl 7-phenoxyacetamido-3- ( l-pyridinomethyl) -t2-cephem-4-carboxylate hydrobromide is formed. The t2-cephalosporan ester product can easily be converted into a known 3-cephalosporanic acid derivative by means of the procedures described above.



   Aliquot portions of a solution of tert-butyl-7- (phen oxyacetamido) -3-bromomethyl-¯2-cephem-4-carboxylate in dry benzene are mixed with 1.1 molar equivalents of one of the amines listed below and for 3 to 5 Stirred for hours at room temperature, the specified cephalosporin ester salts being formed:

  : tert-butyl-7- (phenoxyacetamido) -3- (3'-picolino-1'-methyl) -t2-cephem-4-carboxylate bromide from 3-picoline; tert-butyl-7- (phenoxyacetamido) -3- (4'-carbamyl-1'-pyridino-methyl) -t2-cephem-4-carboxylate bromide from isonicotin amide; tert-butyl-7- (phenoxyacetamido) -3 - (3'-carbamyl-1'-pyridinomethyl) -t2-cephem-4-carboxylate bromide from nicotin amide; tert-Butyl-7- (phenoxyacetamido) -3- [3 '- (N'-methylcarbamyl) -1-pyridino] -methyl-t2-cephem-4-carboxylate bromide from N.
Methylnicotinamide; tert-butyl-7- (phenoxyacetamido) -3- (3'-bromo-N-pyridinomethyl) -t2-cephem-4-carboxylate bromide from 3-bromopyridine;

  ; tert-Butyl-7- (phenoxyacetamido) -3- (2 ', 3'-dimethyl-N-pyridinomethyl) -t2-cephem-4-carboxylate bromide from 2,3-dimethylpyridine; tert. -Butyl-7- (phenoxyacetamido) -3- (3 '-carboxy-N-pyridino-methyl) t2-cephem-4-carboxylate bromide from nicotinic acid; tert. -Butyl-7- (phenoxyacetamido) -3- (4'-cyano-N-pyridinomethyl) -t2-cephem-4-carboxylate bromide from 4-cyanopyridine; tert-butyl 7- (phenoxyacetamido) -3- (3'-sulfonamido-N-pyridinomethyl) -t2-cephem-4-carboxylate bromide from 3-sulfonamidopyridine; tert-butyl 7- (phenoxyacetamido) -3- (1'-pyrimidinomethyl) -t2-cephem-4-carboxylate bromide from pyrimidine;

   tert-Butyl-7- (phenoxyacetamido) -3 - (3'-thiazolylmethyl) -t2-cephem-4-carboxylate bromide from thiazole.



   These t2-cephalosporin ester salts, which are obtainable from penicillin V by means of the Morin-Jackson process and can be converted into t3-cephemic derivatives by means of the method according to the invention, can be processed into antibiotic materials by means of the procedure described above.



   According to the procedure already described, but with the potassium acetate used there being replaced by other oxygen-containing nucleophilic reagents, the specified t2-cephalosporin esters are obtained from the phthalimidomethyl or succinimidomethyl-7- (phenoxyacetamido) -3-bromomethyl-t2-cephem-4-carboxylate intermediates manufactured;

   the following cephalosporin products bearing a functional group in the 3-position are obtained: phthalimidomethyl-7- (phenoxyacetamido) -3- (benzoyloxymethyl) -L2-cephem-4-carboxylate from sodium benzoate; Succinimidomethyl 7- (phenoxyacetamido) -3- (phenylacetoxymethyl) -t2-cephem-4-carboxylate from sodium phenyl acetate; Phthalimidomethyl-7 (phenoxyacetamido) -3- (thiophene-2-carboxymethyl) -t2-cephem-4-carboxylate from sodium thiophene-2-carboxylate; Succinimidomethyl 7- (phenoxyacetamido) -3- (indole-2-acetoxy methyl) -t2-cephem-4-carboxylate from sodium 2-indolacetate;

  ; Phthalimidomethyl 7- (phenoxyacetamido) -3- (carbomethoxy-carboxymethyl) -t2-cephem-4-carboxylate from sodium methyl oxalate; Succinimidomethyl 7- (phenoxyacetamido) 3 (hydroxymethyl) -t2-cephem-4-carboxylate from water;
Phthalimidomethyl 7- (phenoxyacetamido) -3- (methoxycarbo nyloxymethyl) -a2-cephem-4-carboxylate from sodium methyl carbonate; Succinimidomethyl 7- (phenoxyacetamido) -3- (methylthiocarbonyloxymethyl) -t2-cephem-4-carboxylate from sodium S-methyl monothiocarbonate.



   The t2-cephalosporin esters are converted into 3-cephalosporic acid antibiotics by the above-described process of oxidation and reduction.



   1 molar equivalent of 4-methoxybenzyl-7- (phenoxyacetamido) -3-bromomethyl-n2-cephem-4-carboxyl dissolved in absolute methanol was treated with 2 molar equivalents of anhydrous powdered calcium carbonate. The mixture was stirred at room temperature for 24 hours to allow the reaction to proceed, filtered and the filtrate evaporated to dryness. The residue, which contained the 4-methoxybenzyl-7- (phenoxyacetamido) -3-methoxymethyl-A2-cephem-4-carboxylate, was dissolved in benzene. A small amount of pyridine was added to the benzene solution to shift the double bond to form an equilibrium mixture of the n2 and t3 cephalosporin esters.

  After several hours, trifluoroacetic acid in molar excess, based on the product, and anisole were added, whereupon the mixture was stirred for 2 hours, with a mixture of the antibiotic 7-phenoxyacetamido-3-methoxymethyl-, t3-cephem-4-carboxylic acid with 7-phenoxyacetamido-3-methoxymethyl- # 2-cephem-4-carboxylic acid.



   According to the procedure of the above section, but using other oxygen-containing nucleophilic reagents, the following t2-cephalosporin ester products are obtained from the specified nucleophilic reagents: 4-methoxybenzyl-7- (phenoxyacetamido) -3-thiophene-2-methoxymethyl) -t2-cephem-4-carboxylate from thiophene-2-carbinol; 4-Methoxybenzyl-7- (phenoxyacetamido) -3-t4- (tert-butoxy-ca * onylpiperidinyD-oxymethylj-2-cephem-4-carboxylate from 4-hydroxypiperidine-N-tert-butylcarbaminate; 4-methoxybenzyl-7- ( phenoxyacetamido) -3- (diphenylmethyl idenaminooxymethyl) - # 2-cephem-4-carboxylate from benzophenone oxime;

   and 4-methoxybenzyl-7- (phenoxyacetamido) -3- (isopropylideneaminooxymethyl) - # 2-cephem-4-carboxylate from acetone oxime.



   The t2-cephalosporin ester derivatives can easily be converted into the corresponding t3-sulfidic acid antibiotics by means of the procedure described in Example 1 for sulfoxide formation and reduction and subsequent ester cleavage.



   Esterification
A suspension of 6.98 g (0.002 mol) of t2-desacetoxy-ceph-V-acid [3-methyl-7- (phenoxyacetamido) -A2-cephem4-carboxylic acid] and 2.8 g (0.002 mol) of 4-methoxybenzyl alcohol in 100 cm3 of methylene chloride was admixed with a solution of 4.62 g of dimethylformamideineopentyl acetal in 25 cm3 of methylene chloride while stirring. Complete dissolution occurred within a few minutes. The reaction mixture was stirred at room temperature for 48 hours to allow the reaction to proceed to completion. The solvent was removed and benzene added. The resulting solution was washed three times with aqueous sodium bicarbonate solution and twice with aqueous sodium chloride solution, dried over magnesium sulfate, filtered and evaporated.

  The residue was crystallized from carbon tetrachloride and gave 6.74 g (92 / o yield, based on the 2¯ acid not recovered) of 4-methoxybenzyl-3-methyl-7-phenoxyacetamido t2-cephem-4-carboxylic acid ester with a melting point of 108 until 1120 C.



   The consumed aqueous sodium bicarbonate liquids were covered with a layer of ethyl acetate, cooled and brought to pH = 2.5. The organic layer gave 1.56 g of colorless, crystalline 3-methyl-7-phenoxyacetamido- # 2-cephem-4-carboxylic acid, which could be used again for the esterification process.



   Introduction of a functional group on the 3-methyl group
1.17 g of 4-methoxybenzyl-3-methyl-7-phenoxyacetamido-t2-cephem-4-carboxylic acid ester were treated with N-bromosuccinimide in carbon tetrachloride in the presence of azobisisobutyronitrile, with 4-methoxybenzyl-3-bromomethyl-7-phenoxyacetamido , 2-cephem-4-carboxylic acid esters were formed.



   This t2-allyl bromide ester was dissolved in 100 cm3 of absolute methanol which contained 2 molar equivalents of N, N-diethylaniline, in order to absorb the hydrogen bromide formed as a by-product. The mixture was stirred for 24 hours at room temperature to ensure complete conversion, and then evaporated to dryness, taken up in benzene, extracted twice with cold 50% hydrochloric acid solution, sodium bicarbonate solution and sodium chloride solution, dried over magnesium sulfate, filtered and evaporated, whereby 970 mg a brown oil was obtained. This brown oil was chromatographed on a column of silica gel containing 15% water using a mixture of benzene and ethyl acetate as the eluent.

  With a benzene solution containing about 4 O / o ethyl acetate, about 15 O / o unconverted deacetoxy starting material was eluted. 4-Methoxybenzyl-3-m ethoxymethyl-7-Dhenoxyacetamido-t2-cephem-4-carboxylic acid ester with a melting point of 116 to 1180 ° C. (from methanol) was eluted in a yield of 40% with a benzene solution containing about 8/0 ethyl acetate. The structure of the same was confirmed by nuclear magnetic resonance spectrum and elemental analysis.



   Example 3 a) Sulphoxy formation
A cooled solution of 215 mg of 4-methoxybenzyl-3-methoxymethyl-7-phenoxyacetamido-t2-cephem-4-carboxylic acid ester in 5 cc of chloroform was treated with a solution of 85 mg of m-chloropenzoic acid (88 o / o purity) in chloroform . The mixture was stirred and allowed to warm slowly over an hour. The chloroform solution was washed with sodium bicarbonate solution and sodium chloride solution, dried over magnesium sulfate, filtered and evaporated.

  The solid residue was crystallized from benzene containing methylene chloride to give 82 mg of 4-methoxybenzyl-3-methoxymethyl-7-phenoxyacetamido-3-cephem-4-carbo. ylate-1-oxide with a melting point of 183 to 185 C. which gives a satisfactory elemental analysis.

 

   Further experiments have shown that solvents containing hydroxyl groups, such as, for example, isopropanol or tert-butanol, give better results in this reaction than chloroform.



   b) Reduction of the sulfoxide
A solution of 1.028 g (0.002 mol) of 4-methoxybenzyl 3-methoxymethyl-7-phenoxyacetamido-3-cephem-4-carboxylate-1-oxide in 75 cm3 of dry dimethylformamide was mixed with 15 cm3 of acetyl chloride and then with 6 g of sodium dithionite ( Na2S2Q4) added. After stirring at room temperature for 4 hours, the resulting mixture was cooled, and benzene and aqueous sodium bicarbonate solution were added. After the excess acetyl chloride was completely decomposed, water was added and the mixture was extracted twice with benzene.

  The combined benzene extracts were washed with aqueous sodium bicarbonate solution and then with sodium chloride solution and then dried over magnesium sulfate, filtered and evaporated to give 1.32 g of crude, dark brown, semi-solid 4-methoxybenzyl-3-methoxymethyl-7-phenoxyacetamido-n-cephem-4-carboxylate were.



   This crude t3-cephalosporin ester was purified by column chromatography on a column of silica gel containing 15% water and eluted using benzene containing 5% ethyl acetate. 300 mg of 4-methoxybenzyl-5-methoxymethyl-7-phenoxyacetamido-A3-cephem-4-carboxylate with a melting point of 116 to 117.50 ° C. (after crystallization from ethyl ether) were obtained. The assumed structure was verified by nuclear magnetic resonance spectrum and elemental analysis.



   Example 4
Ester cleavage
A solution of 174 mg of 4-methoxybenzyl-3-methoxy methyl-7-phenoxyacetamido-t3-cephem-4-carboxylate and 119 mg of anisole in 25 cm3 of dry benzene was treated with 1.25 cm3 of trifluoroacetic acid. After stirring for 2 hours at room temperature, the reaction mixture was evaporated, the residue was taken up in ethyl acetate and the resulting ethyl acetate mixture was extracted three times with aqueous sodium bicarbonate solution. The aqueous bicarbonate extracts were cooled, covered with ethyl acetate and acidified to pH = 2.8.

  140 mg of a foam were obtained from the ethyl acetate layer by evaporation, which after crystallization from ether gave 86 mg of 3-methoxymethyl-7-phenoxyacetamido-t3-cephem-4-carboxylic acid with a melting point of 135 to 1370.degree. This acid gave an antibiotic active zone on a bioautograph of your paper chromatogram. The stain of the derivative moved slightly more slowly than that of desacetoxyceph-V.



      p-Methoxybenzyl-3-cyanomethyl-7-phenoxyacetamido-2-cephem-4-carboxylate
A portion of crude p-methoxybenzyl-3-bromomethyl 7-phenoxyacetamido / X2-cephem-4-carboxylate obtained from 585 mg of the corresponding p-methoxybenzyl-3-methyl-7-phenoxyacetamido-t2-cephalosporanic acid ester was dissolved in 10 cm3 of dry dimethyl sulfoxide and added to a suspension of 55 mg of cuprocyanide (0.5 molar equivalents, based on the starting material) in 5 cm3 of dimethyl sulfoxide. The mixture was stirred at room temperature for 6 hours.

  Benzene was then added and the resulting mixture was extracted as follows: three times with aqueous sodium chloride solution, twice with cold aqueous 50% hydrochloric acid solution, twice with aqueous sodium bicarbonate solution and twice with aqueous sodium chloride solution. The residue was dried over anhydrous magnesium sulfate, filtered and evaporated to give 392 mg of crude product.



   This crude product was applied to a column of silica gel containing 15% water. The starting material, namely the p-methoxybenzyl-3-methyl-7-phenoxy-acetamido-t2-cephem-4-carboxylic acid ester, was eluted with benzene containing 2% ethyl acetate and benzene containing 4% ethyl acetate. Benzene containing 4 O / o ethyl acetate and benzene containing 8 o / o ethyl acetate gave an eluate which contained the desired p-methoxybenzyl-3-cyanomethyl-7-phenoxyacetamido-3-cephem-4-carboxylate, which after crystallization from ethyl ether contained Melting point 119 to 1200 ° C.

  The structure of this desired product was confirmed by nuclear magnetic resonance, mass spectrometry and elemental analysis.



   Example 5 a) 4-Methoxybenzyl-3-cyanomethyl-7-phenoxyacetamido-, 3-cephem-4-carboxylate-1-oxide
A solution of 372 mg of 4-methoxybenzyl-3-cyanomethyl-7-phenoxyacetamido-t2-cephem-4-carboxylate in 300 cm3 of isopropyl alcohol was treated with a solution of 133 mg of m-chloroperbenzoic acid in isopropyl alcohol. This mixture was stirred at room temperature overnight.



  The sulfoxide which precipitated, namely 4-methoxybenzyl-3-cyano methyl-7-phenoxyacetamido-a3-cephem-4-carboxylate-1-oxide (210 mg, melting point 204-2070 ° C.) was filtered off with suction.



  The filtrate was evaporated to dryness. The residue was taken up in ethyl acetate, washed with aqueous sodium bicarbonate solution, then with aqueous sodium chloride solution and dried over magnesium sulfate, then filtered and evaporated, 160 mg of an oil being obtained from which, after addition of methanol, a further 45 mg of 3-sulfoxy ester crystallized. The structure of 4-methoxybenzyl-3-cyanomethyl-7-phenoxyacetamido-A3-cephem-4-carboxylate-1-oxide was verified by spectroscopic methods.



  b) 4-methoxybenzyl-3-cyanomethyl-7-phenoxyacetamido-t3-cephem-4-carboxylate
A solution of 400 mg of 4-methoxybenzyl-3-cyanomethyl-7-phenoxyacetamido-t, 3-cephem-4-carboxylate-1-oxide in 25 cm3 of dry dimethylformamide was treated with 1.6 g of sodium dithionite (Na2S204) . The mixture was cooled in an ice water bath and 4.5 cm 3 of acetyl chloride was added. The mixture was stirred in the cold bath for 1 hour and then poured into a cold mixture of aqueous sodium bicarbonate solution and benzene.

  After the excess acetyl chloride had completely decomposed, further benzene was added and the mixture was washed with aqueous sodium bicarbonate solution and then with aqueous sodium chloride solution, dried over magnesium sulfate, filtered and evaporated, whereby 620 mg of crude 4-methoxybenzyl-3-cyanomethyl-7-phenoxy acetamido- t3-cephem-4-carboxylate was obtained as the product.



   This crude product was applied to a column containing 40 g of silica gel with 15% water content. The pure 4-methoxybenzyl-3-cyanomethyl-7-phenoxyacetamido-t3-cephem-4-carboxylate was eluted with a mixture of benzene and 4 O / o ethyl acetate and crystallized from ethyl ether. It had a melting point of 145 to 1470 C. The structure was confirmed by means of spectroscopic methods.

 

   Example 6
The 4-methoxybenzyl-3-cyanomethyl-7-phenoxyacetami do-t3-cephem-4-carboxylate was treated at room temperature for 2 hours with a molar excess of trifluoroacetic acid in benzene solution, the corresponding 3-cyanomethyl-7-phenoxyacetamido-t3cephem- 4-carboxylic acid was formed, which has an antibiotic effect against Bacillus subtilis, as a bioautograph of your paper chromatogram shows.



   Production of tert-butyl-7-phenoxyacetamido-3-bromomethyl-A2-cephem-4-carboxylate
A solution of 8.08 g (20 millimoles) of 4-tert-butyl-7 (phenoxyacetamido) -3-methyl-t2-cephem-4-carboxylate, 5.0 g of N-bromosuccinimide and 50 mg of azobisisobutyronitrile in 800 cm3 of carbon tetrachloride was refluxed under nitrogen until the iodine starch test was negative (5 hours).



  The reaction mixture was cooled, the succinimide was removed by filtration and the solvent was distilled off under reduced pressure to give 11.8 g of a crude product. A nuclear magnetic resonance spectrum showed that only tert-butyl-7-phenoxyacetamido-3-bromomethyl-, 2-cephem-4-carboxylate was present, which was contaminated with traces of succinimide (yield <90 O / o).



      tert-butyl 7-phenoxyacetamido-3-isopropoxymethyl-t2-cephem-4-carboxylate
A solution of 3.51 g of 4-tert-butyl-7- (phenoxyacetamido) -3-bromomethyl-t, 2-cephem-4-carboxylate in 100 cm3 of isopropanol was heated on a steam bath for 20 minutes to approx . 700 C heated.



  The excess isopropanol was removed under reduced pressure. The residue was dissolved in benzene, washed with aqueous 50% sodium bicarbonate solution and then with water, dried over sodium sulfate and evaporated to dryness, yielding 2.32 g of crude tert-butyl7-phenoxyacetamido-3-isopropoxymethyl-t2-cephem-carbo - xylate resulted as a product. Spectral analysis proved the assumed structure.



      tert-Butyl-7-phenoxyacetamido-tert-butoxymethyl-t2-cephem-4-carboxylate
A solution of 1.00 g of tert-butyl-7-phenoxyacetamido 3-bromomethyl-t2-cephem-4-carboxylate in 50 cm3 of tert-butyl alcohol was refluxed for 90 minutes (approx.



   830 C) stirred. The alcohol solvent was then removed under reduced pressure and the residue dissolved in ethyl acetate. The ethyl acetate solution was warmed, treated with decolorizing charcoal, filtered and evaporated to dryness to give 1.36 g of crude tert-butyl 7-phenoxyacetamido-3-tert-butoxymethyl-t2-cephem-4-carboxylate as the product. The crude product was chromatographed over 75 g of silica gel with 15 o / o water content and gave 0.563 g of tert. Butyl 7-phenoxyacetamido-3-tert-butoxymethyl-t2-cephem-4-carboxylate as the product obtained in the eluate with benzene containing 10% ethyl acetate. The structure of the product was confirmed by spectral data.



      tert-Butyl-7-phenoxyacetamido-3-cyclopentyloxymethyl-t2-cephem-4-carboxylate was prepared by adding tert. Butyl 7-phenoxyacetamido-3-bromomethyl-, 2-cephem-4-carboxylate in excess cyclopentyl alcohol as described above, heated.



   Similar 3-oxyether-t2-cephalosporin esters are prepared by heating the corresponding 7-acylamido-3-bromo methyl-n2-cephem-4-carboxylate in admixture with a stoichiometric excess of the desired primary, secondary or tertiary alcohol.



   Example 7 a) Oxidation
Using tert-butyl-7-phenoxyacetamido-3-isopropoxymethyl-t2-cephem-4-carboxylic acid ester, this example shows how the 3-oxyether-n2-cephalosporin esters described above can be converted into the corresponding t3-cephem-1- oxydesters are oxidized and reduced to the corresponding t3 cephemesters.



   Crude tert-butyl-7-phenoxyacetamido-3-isopropoxymethyl-t2-cephem-4-carboxylic acid ester (2.32 g) obtained above was dissolved in a mixture of methylene chloride and 50 cm3 of isopropanol and stirred at ice temperature while a solution of 1.00 g of m-chloroperbenzoic acid in 50 cm3 of methylene chloride was added dropwise.



  After the addition was complete, the solvent was removed under reduced pressure. The residue was dissolved in ethyl acetate, washed with aqueous 50% sodium bicarbonate solution, dried over sodium sulfate and evaporated to dryness, yielding 2.16 g of the crude sulfoxydester, namely tert-butyl-7-phenoxyacetamido-3-isopropoxymethyl- t3-cephem-4-carboxylate-1-oxide. The crude sulfoxy ester was chromatographed over 200 g of silica gel with 15% water content and gave 1.10 g of the pure sulfoxy ester, which was eluted with a mixture of 40% ethyl acetate and 60% benzene. The structure of the pure sulfoxy ester was verified by spectral data.



   b) reduction
A solution of 0.891 g of the sulfoxydester tert-butyl 7-phenoxyacetamido-3-isopropoxymethyl, 3-cephem-4-carboxylate-1-oxide, 0.71 g of stannous chloride, 0.5 cm3 of acetyl chloride, 15 cm3 of acetonitrile and 5 cm3 of dimethylformamide was stirred at 250 ° C. for 2 hours. The solvents were removed under reduced pressure and the residue was suspended in ethyl acetate.

  The ethyl acetate suspension was washed twice with aqueous 30% hydrochloric acid solution, aqueous 50% sodium bicarbonate solution and then with water, dried over sodium sulphate and evaporated to dryness, with 0.781 g of the reduced ester (sulphide ester), namely tert.-butyl-7- phenoxyacetamido-3-isopropoxymethyl-t3-cephem-4-carboxylate.



  The structure was verified with the help of spectral analysis.



   Example 8
Ester cleavage
A solution of 0.156 g of tert-butyl-7-phenoxyacetamido 3-isopropoxymethyl-t3-cephem-4-carboxylic acid ester in 7 cm3 of 980% formic acid and water was stirred at 250 ° C. for 25 hours in order to bring the reaction to completion . The reaction mixture was poured into water and extracted with ethyl acetate. The ethyl acetate solution was washed with water and then extracted with aqueous 5% sodium bicarbonate solution. The resulting solution was acidified with aqueous, 30% hydrochloric acid and extracted with ethyl acetate.

  The resulting ethyl acetate solution was dried over sodium sulfate and evaporated to dryness and gave 0.088 g of the free acid, namely 7 phenoxyacetamido3 -isopropoxymethyl-n3-cephem-4-carboxylic acid, which shows in vitro antibiotic activity against a group of penicillin-sensitive and penicillin-resistant staphylococci.



   According to the procedures described above, the following other 3-oxymethyl-, 2-cephem-4-carboxylic acid esters (II) were prepared by adding tert-butyl-7-phenoxyacetamido-3-bromomethyl-A2-cephem-4- carboxylate dissolved and warmed in an excess of the specified alcohol:

   28. tert-Butyl 7-phenoxyacetamido-3-benzyloxymethyl-A2-cephem-4-carboxylate from benzyl alcohol; 29. tert-Butyl-7-phenoxyacetamido-3- (1'-cyclohexylmethoxy) -methyl-t2-cephem-4-carboxylate from cyclohexylmethanol; 30. tert-Butyl 7-phenoxyacetamido-3-n-butoxymethyl-A2-cephem-4-carboxylate from n-butanol; 31. tert-Butyl-7-phenoxyacetamido-3- (2'-tetrahydrofurylmethoxy) -methyl-t2-cephem-4-carboxylate from 2-tetrahydrofurylmethanol; 32. tert-Butyl-7-phenoxyacetamido-3- (acetylmethoxy) -methyl-2-cephem-4-carboxylate from α-hydroxyacetone; 33. tert-Butyl-7-phenoxyacetamido-3- (2'-methoxyethoxy) methyl-2-cephem-4-carboxylate from 2-methoxyethanol;

   34. tert-Butyl-7-phenoxyacetamido-3- (p-methoxybenzyloxy) methyl-t2-cephem-4-carboxylate from p-methoxybenzyl alcohol; 35. tert-Butyl-7-phenoxyacetamido-3-propargyloxymethyl t2-cephem-4-carboxylate from propargyl alcohol; 36. tert-Butyl-7-phenoxyacetamido-3- (2'-trifluoroethoxy) methyl-t2-cephem-4-carboxylate from 2,2,2-trifluoroethanol; 37. tert-Butyl-7-phenoxyacetamido-3- (carboethoxymethoxy) methyl-t, 2-cephem-4-carboxylate from ethyl-a-hydroxy acetate; 38. tert-Butyl-7-phenoxyacetamido-3 - (2'-acetoxyethoxy) methyl n2-cephem-4-carboxylate from 2-acetoxyethanol;

   39. tert-Butyl-7-phenoxyacetamido-3- (2-bromoethoxy) -methyl t2-cephem-4-carboxylate from 2-bromoethanol; 40. tert-Butyl-7-phenoxyacetamido-3- (3'-bromopropoxy) methyl, 2-cephem-4-carboxylate from 3-bromo-n-propanol; 41. tert-Butyl-7-phenoxyacetamido-3 - (2'-allyloxy) -methyl-A2 cephem-4-carboxylate from allyl alcohol; 42. tert-Butyl-7-phenoxyacetamido-3- (2'-methylthioethoxy) methyl-3cephem-4-carboxylate from 2-methylthioethanol; 43. tert-Butyl-7-phenoxyacetamido-3- (4'-nitrophenoxy) methyl-t2-cephem-4-carboxylate from 4-nitrophenol; 44. tert-Butyl-7-phenoxyacetamido-3-phenoxymethyl-A2-cephem-4-carboxylate from phenol;

   45. tert-Butyl-7-phenoxyacetamido-3- (4'-methylphenoxy) -methyl-t2-cephem-4-carboxylate from p-methylphenol; 46. tert-Butyl-7-phenoxyacetamido-3 - (2'-cyanoethoxy) methyl-t2-cephem-4-carboxylate from 2-cyanoethanol; 47. tert-Butyl-7-phenoxyacetamido-3- (3'-tetrahydrofuryloxymethyl) -a2-cephem-4-carboxylate from 3-furyl alcohol; 48. tert-Butyl-7-phenoxyacetamido-3- [1 '- (carboethoxy) -ethoxy] -t2-cephem-4-carboxylate from ethyl 2-hydroxypropionate;

   49. tert-Butyl-7-phenoxyacetamido-3- (2'-chlorallyloxymethyl) -A2-cephem-4-carboxylate from 2-chloroallyl alcohol; 50. tert -Butyl-7-phenoxyacetamido-3 - (2'-thienylmethoxy) -methyl-na-cephem-4-carboxylate from 2'-thienylmethanol.



   The following table contains the nuclear magnetic resonance data corresponding to group 3 of some of the new starting products of formula II. These data are given as size 8 in 10-6 units (p. P. M), based on tetramethylsilane in deuteriochloroform as solvent. The data are given in the following form: a) = chemical shift (ppm) b) = description of the signals of the spectrum: s = singlet d = doublet t = triplet m = multiplet q = quartet c) = number of protons
The first group of compounds were tert-butyl-7 phenoxyacetamido-3-oxymethyl ether-3-cephem-4-carboxylic acid esters of the formula:
EMI14.1


 <tb> S <SEP> GÜ
 <tb> <SEP> --M <SEP> nl
 <tb> <SEP> t <SEP> b1tyi
 <tb> where R had the meanings given.



   R 3-methylene bound to the 3-methylene group -H 4.20 (s) 2.85 (s) (1) -CH3 4.0 (d) 3.29 (s) (3) -CH2CH3 3, 92 (s) 3.30 (q) (2), 1.2 (t) (3)
EMI14.2
 4.05 (s) 3.60 (m) (1), 1.18 (d) (6) 3.95 (s) 1.20 (s) (9) 4.10 (q) 4.50 ( s) (2), 7.35 (s) (5) 4.20 (m) 3.60 (t) (2), 4.20 (t) (2),
2.10 (s) (3) -CH2CH2CN 4.12 (q) 3.60 (t) (2), 2.60 (t) (2) -CH2CH3OCH3 4.20 (q) 3.30 (s) (4), 3.45 (s) (3)
EMI14.3
   4.0-4.2 (m) 4.2 (m) (1) -CH2CH2CH2CH3 4.0 (q) 0.90 (t) (3), 1.50 (m) (4),
3.30 (q) (2)
EMI14.4
 4.10 (m) 1.8 (m) (4), 3.30 (d) (2),
3.8 (m) (3) -CH2CH2SCH3 3.80 (m) 2.1 (s) (3), 3.6 (m) (4)
EMI14.5
 4.20 (m) 1.25 (t) (3), 4.0 (s) (2),
4.2 (q) (2) 4.12 (q) 2.15 (s) (3),

   4.00 (s) (2) -CH2CH2Br 4.10 (q) 3.3-3.9 (m) (4) -CH2ECH 4.17 (s) 2.5 (t) (1), 4, 10 (d) (2) -CH2C8H5 4.0 (q) 3.2 (2) (d), 1.01.9 (m) -CH2CH = CH2 4.0 (m) 4.0 (m) ( 2), 5.0-5.5 (m) (3) -CH2CH2CH2Br 4.10 (m) 2.0 (m) (2), 3.3-3.8 (m) (4)
EMI 14.6
 4.10 (m) 1.0-1.5 (m) (6), 4.2 (m) (3) -CsH5 3.41 (s) 6.7-7.4 (m) Further connections of the Formula:
EMI15.1


 <tb> ¸
 <tb> <SEP> CO2 <SEP> tert0 <SEP> -butyl
 <tb> where X has the following meaning, were also prepared and analyzed by nuclear magnetic resonance spectra.



   X 3-methylene group attached to the 3-methylene group (X) -Br 4.20 (q) -SCH3 3.30 (m) 1.98 (s) (3)
EMI15.2
 3.20 (m) 2.2 (s) (6) -CN 2.01 (s)
Production of tert. -Butyl 7- (phenoxyacetamido) -3 hydroxymethyl-D2-cephem-4-carboxylate
The tert-butyl 7-phenoxyacetamido-3-bromomethyl-n2-cephem-4-carboxylate prepared from 20 millimoles as above prepared 4-tert-butyl-7-phenoxyacetamido-3-methyl-t3-cephem-4-carboxylate was dissolved in 200 cm3 of benzene and treated with 200 cm3 of dimethyl sulfoxide and 5 cm3 of water.



  The reaction mixture was stirred at 250 ° C. for 20 hours. The reaction mixture was then poured into 1000 cc of water containing 20 g of sodium chloride. The benzene layer which separated from the water was then washed four times with water and twice with aqueous 50% sodium bicarbonate solution, dried over sodium sulfate and evaporated to dryness. The residue crystallized from benzene in the form of prisms (3.9 g, 46 o / o) with a melting point of 118 to 1200 C. The spectral data confirmed the proposed structure.



   When this reaction was investigated further, it was found that better yields are obtained with a mixture of benzene and dimethyl sulfoxide in a volume ratio of 50:50 as a diluent for the reaction of this 7-acylamido-3-bromomethyl-a2-cephem-4-carboxylic acid ester with water although good yields are also obtained with mixtures of benzene and 25 to 75 percent by volume of dimethyl sulfoxide. Thin-layer chromatography proved in further experiments that tetrahydrofuran, acetonitrile, ethyl acetate, nitromethane, acetone, dioxane, methyl chloride and similar organic diluents can be used instead of benzene, while dimethylformamide and hexamethylphosphoric acid amide can be used instead of dimethyl sulfoxide.



   Similar 3-hydroxymethyl compounds have been prepared by reacting water with 7-phenoxyacetamido-3-bromo methyl-C2-cephem-4-carboxylic acid esters, which are p-methoxybenzyl, tert-butyl, p-nitrobenzyl and 1 , 1-dimethyl-2-propynyl ester.



   The 3-hydroxymethyl compounds prepared as above can be used to convert the corresponding 3 acyloxymethyl-ft2-cephemesters by reaction with an alkanoic anhydride, e.g. B. acetic anhydride to produce; these can then be converted into known antibiotic compounds by the process described above (oxidation, reduction) and, if necessary, ester cleavage.



   Production of tert-butyl 7- (phenoxyacetamido) -3-dimethylaminomethyl-t2-cephem-4-carboxylate
A solution of crude t2-bromide (tert-butyl-7-phenoxy cetamido-3-bromomethyl-t2-cephem-4-carboxylate) in 50 cmS of ethyl ether was mixed with about 5 cm3 of dimethylamine in 20 cmS of ether at about 0 C offset. The reaction mixture was stirred at 0 ° C. for 3 hours. The solvent was removed under reduced pressure and the residue was suspended in ethyl acetate. The ethyl acetate solution was washed twice with aqueous 50% sodium bicarbonate solution and then extracted with 30% hydrochloric acid.



  The acidic solution was made basic with 5 lo sodium bicarbonate solution and then extracted with ethyl acetate.



  The ethyl acetate solution was dried over anhydrous potassium carbonate and evaporated to dryness to give 2.3 g of crude product. A nuclear magnetic resonance spectrum showed that the pure dimethylamine derivative was present, namely tert-butyl-7-phenoxyacetalmido-3- (N, N-dimethylaminomethyl) -t2-cephem-4-carboxylate.



   A solution of tert-butyl 7-phenoxyacetamido-3-bromomethyl-ft3-cephem-4-carboxylate in dry benzene was mixed with excess methyl mercaptan and subjected to the procedure already described. The product obtained was tert-butyl 7-phenoxyacetamido-3-methylthiomethyl-t2-cephem-4-carboxylate. A nuclear magnetic resonance spectrum gave defined signals for the product, namely a multiplet at 3.3 × 10-o (3.3 ppm) for the 3-methylene group and a singlet at 1.98 × 10-c for the three protons of the thiomethyl group , based on tetramethylsilane in deuterochloroform as solvent standard.

 

   A summary of the properties of process products is given below.
EMI16.1




   Calculated Found Rt R2 mp, OC CHNCHN p -nitrobenzyl CH3 201-205 56.92 4.78 4.97 56.92 4.83 8.05 2-benzothienylmethyl CH3 182-185 60.22 4.87 5.20 60.02 4.97 5.19 3-chlorobenzyl CH3 171-174 58.07 4.87 5.42 57.82 4.91 5.36 3-chlorophenylthiomethyl CH3 138-143 54.68 4.59 5.10 54.84 4.81 5.32
EMI16.2
 CH3 158-165 52.38 4.80 11.11 50.35 5.04 10.00 CH3 149-150 59.77 4.24 15.36 59.70 4.06 5.19 2-Thienylmethyl Et 159- 162 57.37 5.22 5.58 57.64 5.33 5.35
Example 9
Hydrolysis:
From the table above, the p-methoxybenzyl-7 (4'-nitrophenylacetamido) -3-methoxymethyl-a3-cephem-4-carboxylic acid ester was chosen to illustrate this step.



   A solution of 120 mg (0.23 millimoles) of p-methoxybenzyl-7- (4-nitrophenylacetamido) -3-methoxymethyl-3-cephem 4-carboxylate, 0.4 cm3 of trifluoroacetic acid and 20 drops of anisole in 10 cm3 of benzene was added during 1 , Left to stand for 5 hours at room temperature. The solution was then added to 100 cm3 of phosphate buffer (pH = 7). After separating the layers, the aqueous layer was washed with water and saturated sodium chloride solution, dried over magnesium sulfate and concentrated. The residue was dissolved in 15 cm3 of ethanol and 0.4 cm3 of a one-normal solution of sodium acetate in methanol was added.

  After 2 hours at 0 C, the solid material was isolated, washed with ethanol and dried in vacuo at 450 C; it gave 76 mg (83 / o) of the crystalline product, namely 7- (4 'nitrophenylacetamido) -3-methoxymethyl-ns-cephem4-carboxylic acid. The nuclear magnetic resonance and ultraviolet spectra confirmed the structure of this compound. The microanalysis was also in agreement with the calculated values.

  The following table contains a summary of the physical and chemical data of the sodium salts of the above and other acids of the formula given in the table:
EMI17.1

Calculated Found Rt R2 C H N C H N 5
EMI17.2
 CH3 47.54 3.75 9.78 47.70 4.15 9.51 217 (10,800)
265 (16,300) CH3 51.81 3.89 6.36 51.91 3.73 6.30 227 (17,000)
258 (9,550)
297 (2,000) CH3 49.08 4.76 6.02 48.81 4.69 5.73 258 (7,000) CH3 45.28 3.58 6.21 44.99 3.78 5.92 252 (12,300) CH3 41.37 3.72 13.79 41.40 4.01 13.50 CH3 50.99 4.28 7.00 45.56 4.76 6.37 258 (6,800)
Et 57.37 5.22 5.58 57.64 5.33 5.35 CH3 48.89 4.37 7.60 48.60 4.50 7.63
The above new sodium salts of 7-acylamido-3-alcoxymethyl-t3-cephem-4-carboxylic acids are antibiotic effective against a wide variety of gram-positive and gram-negative organisms.



   Example 10 tert -Butyl 7-amino-3-cyanomethyl-3-cephnm-4-carboxylate-p-toluenesulfonate
A solution of 2.15 g (5 millimoles) of tert-butyl-7-phenoxyacetamido-3-cyanomethyl-t3-cephem-4-carboxylate in 100 cm3 of dry benzene was mixed with 590 mg (1.5 equivalents) of dry pyridine and 1.56 g (1.5 equivalent) of phosphorus pentachloride were added. This mixture was heated to 560 ° C. for 2 hours under nitrogen. The benzene was removed and 100 cc of ice cold methanol was added. After standing at room temperature overnight, the methanol was removed under reduced pressure, whereupon 75 cm3 of tetrahydrofuran and, after cooling, 75 cms of cold buffer solution (pH = 4.5) were added.

  After standing for 20 minutes, the tetrahydrofuran was removed under reduced pressure and ethyl acetate and a little water were added. The pH of this mixture was brought to 6.5 with sodium bicarbonate. The organic layer was separated, washed twice with brine, dried over magnesium sulfate, filtered and evaporated to a small volume. A solution of this tert-butyl-7-amino-3-cyanomethyl-t3-cephem-4-carboxylate in 50 cm3 of ethyl acetate was treated with a solution of 951 mg of p-toluenesulfonic acid in 50 cm3 of ethyl acetate. The p-toluenesulfonic acid salt was left to crystallize out overnight and then filtered off with suction; 856 mg with a melting point of 1170 ° C. (decomposition) were obtained.

  The structure was confirmed by infrared, ultraviolet and nuclear magnetic resonance spectra and elemental analysis.



   Example 11 tert -Butyl 7-amino-3-methoxymethyl-t3-cephem-4-carboxylate
A solution of 364 mg (0.84 millimoles) of oily tert. Butyl 7-phenoxyacetamido-3-methoxymethyl, 3-cephem-4-carboxylate in 30 cm3 of dry benzene was mixed with 83 mg of dry pyridine and 218 mg of phosphorus pentachloride (1.25 equivalent). The mixture was heated to 56-580 ° C under nitrogen for 2 hours. The benzene was removed and 35 cc of ice cold methanol was added. This solution was left to stand at room temperature for 24 hours. The methanol was removed under reduced pressure and 20 cm 3 of tetrahydrofuran was added. After cooling, 20 cm3 of pH 4.5 buffer were added, whereupon the mixture was left to stand for 20 minutes at room temperature.

  After removing the tetrahydrofuran under reduced pressure, dilute sodium chloride solution and ethyl acetate were added and the pH was brought to 6.5 with solid sodium bicarbonate. The organic layer was separated, washed twice with saturated brine, dried over magnesium sulfate, filtered and evaporated to small volume. For the purpose of identification, a solution of the tert-butyl-7-amino-3-methoxymethyl-t3-cephem-4-carboxylate in 20 cm3 of ethyl acetate was mixed with a solution of 160 mg of p-toluenesulfonic acid monohydrate.

  The p-toluenesulfonic acid salt of tert-butyl-7-amino-3-methoxymethyl-Q3-cephem-4-carboxylate crystallized spontaneously in an amount of 177 mg (yield 45%). This solid salt was recrystallized from isopropyl alcohol and gave a cleaner sample with a melting point of 183 to 1870 C, the structure of which was confirmed by infrared, ultraviolet and nuclear magnetic resonance spectra and elemental analysis.



   Example 12 7- (2'-Thienyl) -acetamido-3-cyanomethyl-t3-cephem-4-carboxylic acid
A solution of 455 mg of tert-butyl-7-thiophenacetamido 3-cyanomethyl-t3-cephem-4-carboxylate in 40 cm3 of 98-1000 / above formic acid was stirred for 2.5 hours at room temperature under nitrogen. The formic acid was removed under reduced pressure. Ethyl acetate and water were added and the pH was brought to about 7.5 with sodium bicarbonate. Ethyl acetate was added to the aqueous layer, the mixture was cooled and acidified to pH = 2.8 with 200% strength hydrochloric acid.

  The organic layer was removed, washed twice with saturated brine, dried over magnesium sulfate, filtered and evaporated to give 234 mg of a golden colored foam.



   This foamy material was taken up in boiling ethyl ether (10 mg remaining insoluble).



  After evaporation to a small volume, 97 mg of the desired acid, namely 7- (2'-thienyl) -acetamido-3-cyanomethyl-t3-cephem-4-carboxylic acid with a melting point of 114 to 1170 ° C., crystallized. The structure was determined by the infrared spectrum , Ultraviolet spectrum and nuclear magnetic resonance spectrum confirmed. On a bioautograph of a paper chromatogram, the substance showed an antibiotic active zone that moved a little faster than sodium cephalothin.



   Example 13 p-Methoxybenzyl-7-amino-3-methoxymethyl-cephem-4-carboxylate
A solution of 996 mg (2 millimoles) of p-methoxybenzyl 7-phenoxyacetamido-3-methoxymethyl-t3-cephem-4-carboxylate and 221.5 mg (1.4 equivalents) of dry pyridine in 75 cm3 of dry benzene was treated with 601 mg (40 O / o excess) phosphorus pentachloride added. The mixture was heated to 500 ° C. for 2 hours under nitrogen. The benzene was evaporated under reduced pressure and 100 cc of ice-cold methanol was added. After standing at room temperature overnight, the methanol was removed by evaporation under reduced pressure.

  The residue was treated with 40 cm3 of tetrahydrofuran and, after cooling, with 40 cm3 of water. This homogeneous mixture was left to stand for 25 minutes; the tetrahydrofuran was removed under reduced pressure, ethyl acetate and a little water were added and the pH was brought to 6.5 with solid sodium bicarbonate. The organic layer was separated, washed twice with saturated brine, dried with magnesium sulfate, filtered and evaporated to a small volume.



  For identification purposes, a solution of the crude p-methoxybenzyl-7-amino-3-methoxymethyl ti3-cephem-4-carboxylate obtained therefrom in a total of 80 cm3 of ethyl acetate with a solution of 380 mg (2 millimoles) of p-toluenesulfonic acid monohydrate in 10 cm3 ethyl acetate added. The p toluenesulfonic acid salt began to crystallize out within a few minutes. After 3 hours the crystals were filtered off with suction; 781 mg (73 0 / o) crystals with a melting point of 160 to 1640 ° C. were obtained. The structure was confirmed by infrared, ultraviolet and nuclear magnetic resonance spectra and elemental analysis.



   Example 14
7- [2- (sydnone-3) -acetamidoj-3-methoxymethyl- / S3-cephem-4-carboxylic acid
86 mg of tert-butyl 7- [2- (sydnon-3) acetamino] -3-methoxymethyl-t3-cephem-4-carboxylate were dissolved in 2 cm 3 of 980 / above formic acid and stirred at room temperature for 2 hours . The solution was evaporated to dryness. The semi-solid residue that remained was triturated with ethyl ether and the solid substance isolated and dried in vacuo, giving 52 mg of 7- [2- (sydnone-3-) acetamino] -3-methoxymethyl-t3-cephem-4-carboxylic acid received. Elemental microanalysis and ultraviolet analysis were consistent with the stated structure.



   Example 15
The tert-butyl-7- (2'-carboxy-2'-phenylacetamido) -3-methoxymethyl-t3-cephem-4-carboxylic acid ester from Example 62 was stirred with 1 cm3 of cold trifluoroacetic acid. After 15 minutes the solution was evaporated. The residue was stirred for 1 hour in 5 cm3 of a mixture of petroleum hydrocarbon (boiling range 60 to 680 C) and ethyl ether in a volume ratio of 1: 1 and then isolated and dried in vacuo, with 56.1 mg of 7- (2'-carboxy 2'-phenylacetamido) -3-methoxymethyl-t3-cephem-4-carboxylic acid were obtained. The elemental analysis and the ultraviolet spectrum were in agreement with this structure.

 

   The above new cephalosporins gave characteristic Rf values in paper chromatography and were microbiologically tested in vitro, with all compounds showing excellent activity against various staphylococcal strains. Oral administration of these derivatives to mice protected the animals from death from streptococcal infections.



   Example 16 7-Amino-3-methoxymethyl- # 3-cephem-4-carboxylic acid
To 268 mg (1 millimole) of the p-toluenesulfonic acid salt of p-methoxybenzyl-7-amino-3-methoxymethyl- # 3-cephem-4-carboxylate, which were stirred in water, sodium orthophosphate was added until pH = 7, while the Solution was overlaid with ethyl acetate. The ethyl acetate solution was dried over magnesium sulfate and evaporated to dryness. The oily residue was dissolved in 25 cm3 of benzene containing 40 drops of anisole and 0.8 cm of trifluoroacetic acid. After stirring the mixture for 2 hours at room temperature, the benzene was evaporated and the residue was taken up in 10 cm of water and covered with a layer of 10 cm of ethyl acetate. The pH was brought to 3.6 by adding triethylamine.

  After cooling the mixture overnight, the solid material was isolated, washed with water and ethyl acetate and dried in vacuo at 450 ° C. to give 75 mg of 7-amino-3-methoxymethyl- # 3-cephem-4-carboxylic acid.



  The structure was confirmed by infrared, ultraviolet and nuclear magnetic resonance spectrum.



   A similar procedure was used to prepare 7-amino-3-ethoxymethyl- # 3-cephem-4-carboxylic acid from its p-methoxybenzyl ester.



   The following table summarizes the antibiotic data obtained with the solutions of the compounds with the indicated residues R and the Rf values obtained with the bioautograms.
EMI19.1




  Example R G + G + G + Tb G- G- G- G
Rk * SA X-12 X-186 X-85 X-45 X-142 X-161 X-48
70 CHa (CH2) s 1.1 32 20 30 - 11 - - -
71 CHa (CH2) 4-CH = CH- 1.5 27 22 32 - 12 - -
EMI19.2
 0.8 24 26 31 - 14 - - - 0.9 29 28 35 - 17 - track 2.2 30 29 45 = 13 - - - 75 CHs (CH2) sSCH2- 1.0 30 33 40 - 14 - 16 - 76 (CHs) 2CH (CH2) 2SCH2- 1.4 35 34 39 - 16 - 16 - 77 CH2 = CHCH2SCH2- 0.9 25 21 29 - 17 trace - 78 p-CHsO-CsH4-CH2- 0.8 29 34

   40 - 16 - 15 79 p-NO2-CoH4-CH2CH2- 1.0 32 30 37 - 12 - 11 - 80 m-CI-CGH4-OCH2- 1.3 36 34 42 - 15 - 13 81 p-Cl-CGH4 -OCH2- 1.1 40 35 37 - 13 - trace 82 m-NO2-CGH4-OCH2- 1.0 35 35 40 - 12 - 12 83 m-CF3-CeH-OCH3- 1.8 34 34 40 - 12 - Lane 84 CGH4-CH2OCH2- 1.0 27 25 32 - 14 - - - 85 pF-CeH4-CH2SCH2- 1.3 35 32 45 lane 15-14 86

   C6H5-CH2CH2SCH2- 1.4 29 34 40 - 14 - 16 - 87 C6H5- (CH2) 3SCH2- 1.7 31 34 37 - 10 - 13 - 88 CeH5-CH = CH- 1.0 25 17 20 - - - - -
Comments * The Rli value is defined as the ratio between the distance of the relevant spot in the bioautogram from the starting point and the distance between the spot of the standard antibiotic, namely sodium cephalothin, and the starting point.



   Bioautograms of paper chromatograms of the test cephalosporin compound and the standard compound (sodium cephalothin from Eli Lilly, brand name Keflin, which is commercially available and has antibiotic activity) were used to determine the Rk values. The method of Bird and Pugh in Antibiotics and Chemotherapy, Volume IV, No. 7 (July 1954), but using Whatman No. 4 paper, which was buffered to pH 4.6 with 0.1 molar sodium acetate, a mixture of 600 parts of methyl ethyl ketone and 400 parts of water as the chamber fluid, and the developing solvent a mixture of 92 parts of methyl ethyl ketone and 8 parts of water was used. Each chromatogram was run for 2.5 to 3.5 hours.



   Paper chromatography techniques are generally described in Paper Chromatography, 3rd Edition, edited by I. M. Hais et al., Academic Press, New York and London 1963, p. 155.



   Furthermore, the zone sizes are given in mm which were obtained in an agar plate test with the following microorganisms: 5. A. = Staphylococcus aureus X12 = B. subtilis X186 = Sarcina sp.



  X85 = Sacharomyces pastorianum X45 = Proteus vulgaris X142 = Salmonella gallinarum X161 = Escherichia coli X48 = Pseudomonas aeruginosa
If a zone size of more than 8 mm is obtained, this is an indication of the antibiotic activity of the test compound against the test organism in this qualitative test.



   PATENT CLAIM 1
Process for the preparation of a cephem derivative
EMI20.1
 in which R is hydrogen, the group
EMI20.2
 where X denotes the amino or a protected amino group, the hydroxyl group, a (C1-Cs) alkoxy group, the carboxyl group or a (Ci-Cs) alkanoyloxy group, the group
EMI20.3
 where X is oxygen, sulfur or a single bond, y is 0 or an integer from 1 to 2 and z is an integer from 1 to 3, or the group
EMI20.4
 and Q in each of the above groups is hydrogen, an alkyl or alkoxy group with 1 to 3 carbon atoms, the cyano, nitro, hydroxy or trifluoromethyl group, a chlorine, bromine or fluorine atom, an a amino (ClC3 ) alkyl group, the carboxymethyl or carbamoylmethyl group,

   or the group
EMI20.5
 where Y is sulfur or oxygen and each of the symbols Z is hydrogen or both Z together with the carbon atoms to which they are attached form a benzene ring, or the group
EMI20.6
 where each of Rt and R2 is hydrogen or the methyl group, or a sydnone-3-carbonyl, 2- (sydnon-3) -acetyl or 2- (sydnon-3) -propionyl group, a (Ct to Cto) alkanoyl group, a ( C-Cio) alkenoyl group, a (C9-C7) alkyl-X- (C1-Cs) alkanoyl group, where X is oxygen or sulfur, a (C3-C7) alkenyl-X- (Ct to C3) alkanoyl group, where X is oxygen or sulfur, a (Cz-Clo) haloalkanoyl group, where the halogen is a fluorine,

   Is a chlorine or bromine atom, or a (C2-Clo) -cyanoalkanoyl group, R 'denotes the cyano group, the group OY, where Y is a (Ci-Cto)' alkyl group, a (C-Cto) alkenyl group, a (C8-Cto ) - Alkynyl group, a (Cs-C7) cycloalkyl group, a (C5 to C7) cycloalkyl- (CtC3) alkyl group, a (C2Cto) haloalkyl group, a (C-Cto) haloalkenyl group or a (C3-Cto) haloalkynyl group, where the halogen is a fluorine, chlorine or bromine atom, a (Ct-C3) -alkyl-X- (C-Cs) alkyl group, where X is oxygen or sulfur,

   a (Ct-C) alkanoyloxy (C-Ce) alkyl group, a (C2-C4) alkanoyl (Ci-Ce) alkyl group, a furyl (Cr

** WARNING ** End of DESC field could overlap beginning of CLMS **.



   

 

Claims (1)

** WARNING ** Beginning of CLMS field could overlap end of DESC **. manure and the standard compound (sodium cephalothin from Eli Lilly, brand name Keflin, which is commercially available and has antibiotic activity) is used. The method of Bird and Pugh in Antibiotics and Chemotherapy, Volume IV, No. 7 (July 1954), but using Whatman No. 4 paper, which was buffered to pH 4.6 with 0.1 molar sodium acetate, a mixture of 600 parts of methyl ethyl ketone and 400 parts of water as the chamber fluid, and the developing solvent a mixture of 92 parts of methyl ethyl ketone and 8 parts of water was used. Each chromatogram was run for 2.5 to 3.5 hours. Paper chromatography techniques are generally described in Paper Chromatography, 3rd Edition, edited by I. M. Hais et al., Academic Press, New York and London 1963, p. 155. Furthermore, the zone sizes are given in mm which were obtained in an agar plate test with the following microorganisms: 5. A. = Staphylococcus aureus X12 = B. subtilis X186 = Sarcina sp. X85 = Sacharomyces pastorianum X45 = Proteus vulgaris X142 = Salmonella gallinarum X161 = Escherichia coli X48 = Pseudomonas aeruginosa If a zone size of more than 8 mm is obtained, this is an indication of the antibiotic activity of the test compound against the test organism in this qualitative test. PATENT CLAIM 1 Process for the preparation of a cephem derivative EMI20.1 in which R is hydrogen, the group EMI20.2 where X denotes the amino or a protected amino group, the hydroxyl group, a (C1-Cs) alkoxy group, the carboxyl group or a (Ci-Cs) alkanoyloxy group, the group EMI20.3 where X is oxygen, sulfur or a single bond, y is 0 or an integer from 1 to 2 and z is an integer from 1 to 3, or the group EMI20.4 and Q in each of the above groups is hydrogen, an alkyl or alkoxy group with 1 to 3 carbon atoms, the cyano, nitro, hydroxy or trifluoromethyl group, a chlorine, bromine or fluorine atom, an a amino (ClC3 ) alkyl group, the carboxymethyl or carbamoylmethyl group, or the group EMI20.5 where Y is sulfur or oxygen and each of the symbols Z is hydrogen or both Z together with the carbon atoms to which they are attached form a benzene ring, or the group EMI20.6 where each of Rt and R2 is hydrogen or the methyl group, or a sydnone-3-carbonyl, 2- (sydnon-3) -acetyl or 2- (sydnon-3) -propionyl group, a (Ct to Cto) alkanoyl group, a ( C-Cio) alkenoyl group, a (C9-C7) alkyl-X- (C1-Cs) alkanoyl group, where X is oxygen or sulfur, a (C3-C7) alkenyl-X- (Ct to C3) alkanoyl group, where X is oxygen or sulfur, a (Cz-Clo) haloalkanoyl group, where the halogen is a fluorine, Is a chlorine or bromine atom, or a (C2-Clo) -cyanoalkanoyl group, R 'denotes the cyano group, the group OY, where Y is a (Ci-Cto)' alkyl group, a (C-Cto) alkenyl group, a (C8-Cto ) - Alkynyl group, a (Cs-C7) cycloalkyl group, a (C5 to C7) cycloalkyl- (CtC3) alkyl group, a (C2Cto) haloalkyl group, a (C-Cto) haloalkenyl group or a (C3-Cto) haloalkynyl group, where the halogen is a fluorine, chlorine or bromine atom, a (Ct-C3) -alkyl-X- (C-Cs) alkyl group, where X is oxygen or sulfur, a (Ct-C) alkanoyloxy (C-Ce) alkyl group, a (C2-C4) alkanoyl (Ci-Ce) alkyl group, a furyl (Cr to C3) alkyl group, a thienyl (C1-Cs) alkyl group, the 3 tetrahydrofuryl group, a tetrahydrofuryl (Ci-Cs) alkyl group, the phenyl, benzyl or phenylethyl group or one on the phenyl ring by a (Ct-C) Alkyl group, (C1 to C3) alkoxy group, (Ci-Ca) alkoxycarbonyl group, the fluorine, chlorine or bromine atom, the nitro, cyano or trifluoromethyl group, substituted phenyl, benzyl or phenylethyl group and R "denotes hydrogen, one equivalent of one pharmaceutically acceptable cation, a (C4-Ce) -tert.-alkyl group, a (Cs-C,) tert.alkenyl group, a (Cs-C7) tert.alkynyl group, a benzyl, methoxybenzyl, nitrobenzyl, Benzhydryl, phthalimidomethyl, succinimidomethyl, trimethylsilyl or phenacyl group, or an inner salt of a compound of the formula I with a free carboxyl group or an acid addition salt of a compound of the formula I, characterized in that a corresponding compound of the formula: EMI21.1 or an inner salt of a compound of the formula II having a free carboxyl group is converted into the corresponding t3 sulfoxide with an oxidizing agent and this is then reduced. SUBCLAIMS 1. The method according to claim I, characterized in that a compound of the above formula in which R is hydrogen, R 'is cyano and R "is a (C-Ce) tert-alkyl group, or an acid addition salt of this compound with an aromatic ( Cs-Cla) sulfonic acid. 2. The method according to dependent claim 1, characterized in that 7-amino-3-cyanomethyl-t3-cephem-4-carboxylic acid tert-butyl ester or the p-toluenesulfonic acid salt thereof is prepared. 3. The method according to claim I, characterized in that a compound of the above formula in which R is hydrogen, R 'is the group OY, Y is a (C1-Clo) alkyl group and R "is a (C-Ce) tert-alkyl group mean, or an acid addition salt of this compound with an aromatic (Ce-Clz) sulfonic acid. 4. The method according to dependent claim 3, characterized in that 7-amino-3-methoxymethyl-t3-cephem-4-carboxylic acid tert-butyl ester or the p-toluenesulfonic acid salt thereof is prepared. 5. The method according to claim I, characterized in that a compound of the above formula is prepared in which R is a sydnone-3-carbonyl, 2- (sydnone-3) acetyl or 2- (sydnone-3) propionyl group, R 'signifies the group OY, Y signifies a (Ct-Cio) alkyl group and R "signifies hydrogen. 6. The method according to dependent claim 5, characterized in that 7- [2- (sydnone-3) acetamino] -3-methoxy methyl-t3-cephem-4-carboxylic acid is prepared. 7. The method according to claim I, characterized in that a compound of the above formula in which R is the group EMI21.2 where X is an amino or protected amino group, a (Ct-C) alkoxy group, the carboxyl group or a (Ct to Cs) alkanoyloxy group and Q is hydrogen, R 'is the group OY, Y is a (Ci-Cto) alkyl group and R "What - mean hydrogen, or the sodium or potassium salt thereof. 8. The method according to dependent claim 7, characterized in that 7- (D-2'-amino-2'-phenylacetamino) -3-methoxymethyl-t3-cephem-4-carboxylic acid is prepared. 9. The method according to dependent claim 7, characterized in that 7- [D-2'-amino-2 '- (3 "-hydroxyphenyl) acetamino] -3-methoxymethyl-t3-cephem-4-carboxylic acid is prepared. 10. The method according to dependent claim 7, characterized in that 7- [D-a-hydroxy-a-phenylacetamino] -3-methoxymethyl-as-cephem-4-carboxylic acid or the sodium or potassium salt thereof is prepared. 11. The method according to claim I, characterized in that the 3-cephem-4-carboxylic acid esters obtained are converted into corresponding t3-cephem-4-carboxylic acids by hydrolysis. 12. The method according to dependent claim 11, characterized in that the hydrolysis is carried out by treatment with trifluoroacetic acid or with formic acid or acetic acid. PATENT CLAIM II Use of the 7-acylamino-group-containing compounds obtained by the process according to claim I for the preparation of the corresponding 7 amino compounds, characterized in that said process products are treated in the presence of a tertiary amine with phosphorus pentachloride, then with an alcohol and finally with water.