NO743667L - - Google Patents

Description

"Antibacterial cephem derivatives and pro-

method of manufacture thereof"

The present invention relates to cephem derivatives.

The compounds are antibacterial agents which are valuable as animal fur additives, as therapeutic agents for the control of infectious diseases caused by gram-positive and gram-negative bacteria, and for sterilizing hospital surfaces, etc. and new intermediates for their production. More particularly, the antibacterial compounds according to the present invention are acylated derivatives of 7-amino-3-substituted-A 3-cephem compounds containing a 5-tetrazolyl group in the 4-position.

Despite the large number of cephem derivatives that have been proposed as antibacterial agents, there still exists a need for new agents.

The antibacterial compounds according to the invention, and the intermediates from which they are produced by acylation, are new compounds, and they could not be predicted from what was known in the field. In U.S. patents 3,427,302 and 3,468,874 penam derivatives are described which comprise a tetrazolyl group as part of the 6-acylamino substituent and in Japanese patent no. 71-38503 cephem derivatives are described which comprise a tetrazolyl group as part of the 7-acylamino substituent. Cephem derivatives with a tetrazolyl-thiomethyl group in the 3-position are also known compounds (U.S. Patent No. 3,641,021). The compounds according to the present invention are characterized in that they have a tetrazolyl group attached directly to the cephem nucleus.

An overview of the biological and non-biological applications of tetrazoles is given by Benson, "Heterocyclic Compounds", Elderfield, Ed., Vol. 8, John Wiley&Sons, Inc., New York,

N.Y., 1967, ch. 1 while a collection of cephem references is set forth in U.S. Patent No. 3,766,176.

It has now been shown that certain 7-acylamino-3-substituted-4-(tetrazol-5-yl)-A 3-cephem derivatives and salts thereof are useful as antibacterial agents/while certain 7-substituted amino-3-substituted- 4-carbamoyl and 4-(tetrazol-5-yl)-A3* and -A<2->cephem derivatives are useful as intermediates for the preparation of these antibiotics.

A preferred group of compounds useful as intermediates are those having the formula

wherein R is hydrogen or an amino protecting group selected from the group consisting of 2,2,2-trichloroethoxycarbonyl, 2,2,2-tribromomethoxycarbonyl, benzyloxycarbonyl and wherein R3, and Rg are each hydrogen, chlorine, bromine, fluorine, methyl, methoxy or phenyl" A is hydrogen, acetoxy, 1-methyltetrazolylthio or 2-methyl-1,3,4-thiadiazolyl-5-thio, and R2 is a potential tetrazole protecting group selected from the group consisting of

where Rg is hydrogen, alkyl containing from 1 to 3 carbon atoms or phenyl, R^ is hydroxy, methoxy, alkanoyloxy containing

from 2 to 4 carbon atoms or benzyloxy and Rq is hydrogen, hydroxy, fluorine, chlorine, bromine, iodo, methyl, methoxy, alkanoyloxy containing from 2 to 4 carbon atoms, phenyl or benzyloxy and

where Rg and R1Q are each hydrogen or methyl and X is oxygen or sulfur. Another preferred class of compounds are intermediates of the formulas and salts thereof where R is hydrogen or an amino protecting group selected from the group consisting of 2,2,2-trichloroethoxycarbonyl, 2,2,2-tribromomethoxycarbonyl, benzyloxycarbonyl and

where R3, R4 and Rg are each hydrogen, chlorine, bromine, fluorine, methyl, methoxy or phenyl, A is hydrogen, acetoxy, 1-methyl-5-tetrazolyl-thio or 2-methyl-1,3,4-thiadiazolyl- 5-thio, ^ is hydrogen, alkanoyloxymethyl containing from 3 to 6 carbon atoms,

1-(alkanoyloxy)ethyl containing from 4 to 7 carbon atoms, methoxymethyl or phthalidyl, and R^ is hydrogen, alkanebyloxymethyl containing from 3 to 6 carbon atoms, 1-(alkanoyloxy)ethyl containing from 4 to 7 carbon atoms, methoxymethyl, phthalidyl or a potential tetrazole protecting group selected from the group consisting of

where Rg is hydrogen, alkyl containing from 1 to 3 carbon atoms or phenyl, R? is hydroxy, methoxy, alkanoyloxy containing from 2 to 4 carbon atoms or benzyloxy and Rg is hydrogen, hydroxy, fluorine, chlorine, bromine, iodine, methyl, methoxy, alkanoyloxy containing from 2 to 4 carbon atoms, phenyl or benzyloxy and where R9 and R1Q each is hydrogen or methyl and X is sulfur or oxygen• A third class of preferred cephem derivatives and salts thereof are those having the formulas

where Ar is cyano, bromo, phenyl mono- or disubstituted phenyl where each substituent is hydroxy, fluoro, chloro, bromo, amino, methoxy or methyl, phenoxy, phentylthio, pyridylthio, thienyl, 2-methyl-1,3,4-thiadiazole -5-ylthio or 1-tetrazolyl, Q is hydrogen, hydroxy, azide, amino or carboxy, n is an integer from 0 to 1,

A is hydrogen, acetoxy, 1-methyltetrazol-5-ylthio or 2-methyl-1,3,4-thiadiazol-5-ylthio, and R. is hydrogen, alkanoyloxymethyl Containing from 3 to 6 carbon atoms, 1-(alkanoyloxy) ethyl containing from 4 to 7 carbon atoms, methoxymethyl or phthalidyl, provided that when Ar is pyridylthio, phenoxy, phenylthio, 2-raethyl-1,3,4-thiadiazol-5-ylthiomethyl, cyano or bromo and n is equal to 1 , Q is hydrogen or carboxy.

The present invention also includes compounds of the third class where A, Q,n and R^ are as indicated and Ar is selected from the group consisting of hydrogen, alkyl containing from 1 to 12 carbon atoms, alkenyl containing from 2 to 12 carbon atoms, cycloalkyl containing from 3 to 7 carbon atoms, cycloalkenyl containing from 5 to 8 carbon atoms, cycloheptatrienyl, 1,4-cyclohexadienyl, 1-aminocycloalkyl containing from 4 to 7 carbon atoms, 5-methyl-3-phenyl-4-isoxazolyl, 5 -methyl-3-(o-chlorophenyl)-4-isoxazolyl, 5-methyl-3-(2,6-dichlorophenyl)-4-isoxazolyl, 5-methyl-3-(2-chloro-6-fluorophenyl)-4 -isoxazolyl, 2-alkoxy-l-naphthyl where the alkoxy group contains from 1 to 4 carbon atoms, oydhonyl, furyl, pyridyl, thiazolyl, isothiazolyl, pyrimidinyl, triazolyl, imidazolyl, pyrazolyl, substituted phenoxy, substituted phenylthio, substituted pyridylthio, substituted thienyl, substituted furyl, substituted pyridyl, substituted tetrazolyl, substituted thiazolyl, substituted isothiazolyl, substituted rt pyrimidinyl, substituted triazolyl, substituted imidazolyl and substituted pyrazolyl, each substituted group may be substituted with up to two groups selected from the group consisting of fluorine, chlorine, bromine, hydroxy, hydroxymethyl,

amino, N,H-dialkylamine where each alkyl group contains from 1 to 4 carbon atoms, alkyl containing from 1 to 4 carbon atoms,

aminomethyl, aminoethyl, alkoxy containing from 1 to 4 carbon atoms, alkylthio containing from 1 to 4 carbon atoms, 2-aminoethoxy and N-alkylamino containing from 1 to 4 carbon atoms.

As those skilled in the art will readily appreciate, the ct carbon atom of the antibacterial oephem side chain to which the amino or hydroxy, (Q) group is attached is an asymmetric carbonatoin which allows the existence of two optically active isomers, the D and L diastereoisomers as well as the racemate , the DL form. According to what has been previously found regarding the activity of such cephem compounds having asymmetric α-carbon atoms, compounds of the present invention having the D configuration are more active than those having the L configuration and are the preferred compounds, although The L and DL forms of the compounds are also covered by the present invention.

It is further worth noting that in the case of asymmetric centers, there are several in the A -céfem core, the basic element from which the compounds of the present invention are derived. These potential additional isomers are not significant in this connection since the 7-amino-A 3-oephera-4-carboxylic acid used leading to the products of the invention is that produced by fermentation and is of one configuration.

Similarly, the term "tetrazole protecting group" or "tetrazolyl-cefera-nitrogeh-protecting group" is intended to mean. all known groups, or which are obvious to the person skilled in the art, can be used (a) to allow the synthesis of compounds where R is an amino-protecting group' and R is the tetrazolylcephem-nitrogen-protecting group, following the procedure starting from 7-( protected amino)cephem-4-carboxylic acid as set forth below" and (b) can be removed from the compound wherein R 2 is the tetrazolyl-cepheity nitrogen-protecting group, wherein R is selected from the group

which consists of hydrogen and an amino-protecting group, and R 2 is the tetrazolylcepheme-nitrogen-protecting group, by using a method in which the cepheme ring system remains substantially intact. It is likewise the ability of the tetrazolylcephem nitrogen protecting group to perform a given function, which will be discussed in more detail below, rather than its exact chemical structure, which is important, and the novelty of the antibacterial agents of the present invention does not depend of the structure of the protecting group. Selection and determination of the suitable protecting groups can easily and simply be done by the person skilled in the art, and examples of a number of applicable groups are given below.

One embodiment according to the invention comprises a method for producing compounds where the araino group in the 7-position is aoylated.

Another embodiment according to the invention comprises the preparation of compounds where the amino group in the 7-position is protected with one amino-protecting group by the transfer of the amide in the 4-position to a tetrazolyl group, protecting the amide and the resulting tetrazolyl group with a tetrazolyl cephem nitrogen-protecting group.

The present invention relates to certain new compounds and new preparations which are usable as antibacterial agents, and as intermediate products for the production of these. For the sake of simplicity, the compounds are designated as derivatives of A 3 -cephem. The term "A 3-cephem" is given by Morin et al. in Journal of

the American Chemical Society, 84, 3400 (1962) and means the structure

Using this terminology, the well-known antibiotic cephalosporin C is designated as 7-(5-amino-5-carboxyvaleramido)-3-acetoxy-3

methyl-A-cephem-4-carboxylic acid.

Many of the compounds according to the present invention are also 5-substituted tetrazoles which can exist in two isomeric forms, namely

Those skilled in the art will understand that when the substituent L means hydrogen, the two forms exist simultaneously in a dynamic, tautomeric, equilibrium mixture. However, in the case where L is different from hydrogen, the two forms represent different chemical structures that do not spontaneously intermingle.

According to the method used to synthesize the cephem intermediates and the antibacterial agents of the present invention, there are two preparative routes, the first being illustrated as follows where R, A, X^, Ar, n and Q are as indicated previously and R^ and R ^ each is hydrogen.

7-carboxamido-3-substituted-'A 3-cephem-4-carboxylic acids (1), prepared experimentally by acylation of the corresponding 7-amino-cephem compound, are transferred to 4-carbamoyl compounds (2) by reacting the 4-carboxy group, activated as 2,4-dinitrophenol ester, with a suitable amine, R^H,,.

The preparation of the compounds 2 where R is derived from triphenylmethyl is achieved either by alkylation of the 7-amino compound, followed by the formation of the 4-carbamoyl group as mentioned above, or by selectively removing the R-acyl group in compounds with the structure 2, such as removing the 2,2,2-trihaloethoxycarbonyl group using acetic acid and zinc dust, and then alkylating the 7-amino-4-carbamoylcepheme compound with the denatured triphenylmethyl chloride.

The conversion of 2 to 3 requires transfer of the 4-carbamoyl group in 2 to the appropriate imino chloride, followed by conversion of this compound to azide. The formation of the iminochloride is conveniently carried out by using phosgene or phosphorus pentachloride in a reaction-inert solvent such as chloroform, while the reaction of the iminochloride with the salt of hydrazoic acid and tetramethylguanidine leads to the formation of the tetrazole ring. As the person skilled in the art will understand, many sources of azide can be used in this reaction and these sources include salts of hydrazonic acid with inorganic bases such as sodium azide, lithium azide, potassium azide and ammonium azide. Since many metal azides are explosive, it is advantageous, and

in this case it is preferred that azides formed from organic bases are used} tetramethylguanidine hydrogen azide is particularly suitable for this purpose.

The sequence step for the transfer of 3> to A requires the removal of "the amino protecting group", R. The reaction conditions used for the removal are given by the type of group to be removed. As mentioned earlier, the 2,2,2-trihaloethoxycarbonyl group can be conveniently removed by using zinc dust and acetic acid, the triphenylmethyl group is removed by using formic acid and the benzdoxycarbonyl group is removed by treating 3^ with a mixture of trifluoroacetic acid/anisole (4 µl v/v ) and trifluoromethylsulfonic acid.

In the latter method, it is preferred that the reaction is carried out at ice bath temperature (0°C) and during a limited time, usually 4-6 minutes. If high temperatures are used, such as 25-40°C, or along reaction times, such as 1-3 hours, it is possible to remove the "tetrazole-blocking" group at the same time.

After the removal of the "amino-protecting" group, the variable R 2 is removed by treating the 4- or p-toluenesulfonyl salt thereof with the previously mentioned mixture of trifluoroacetic acid/anisole.

Aoylation of IS with the appropriate carboxylic acid which is

activated either as an acid halide, activated ester, mixed anhydride or the acid with a carbodiimide give the antibacterial compounds according to the present invention.

Those skilled in the art will readily appreciate that the presence of other functional groups in the acylating acid may require that the groups be masked to prevent them from participating in competing reactions.

When the acylation is finished, the groups can be removed.

In the preparation of compounds with the structure 6,

where Q is amino, it is e.g. necessary that the amino group is blocked, preferably with a t-butoxycarbonyl group, the blocking group being removed by acid treatment after the acylation has ended. A similar practice is required when Q is hydroxy, in which case a formyl group is used to mask the hydroxy group.

The compounds with the structure 6 where Ar is bromine, n is equal to 1 and Q is hydrogen can, in addition to their antibacterial effect, be reacted with mercaptans and additional antibacterial compounds are obtained.

The starting materials for the sequence of reactions are either readily available as commercial reagents or can be prepared according to methods indicated in the literature. The 7-amino-3-substituted-A<3->cephem-4-carboxylic acids are e.g. disclosed in U.S. Patent 3,641,021 and the amines R 2 NIi 2 ^rems:t^lles suitably by one or more of the methods set forth by Wagner and Zook, "Synthetic Organic Chemistry", John Wiley and Sons, Inc., New York, NY, 1956, chapter 24, pp. 653-727" while the triphenylmethyl chlorides used can be prepared according to methods indicated by Bachmann, Org. Synthesis, 23, 100 (1943).

The other method applicable to the synthesis

of the antibacterial compounds according to the present invention and for the production of the intermediate products is illustrated as follows:

where R, A, R 2 , Ar, a and Q are as previously indicated and R 2 is methoxymethyl.

The 7-acylamino-3-methyl-A-cefera-4-carboxylic acid 1 isomerizes to the corresponding 7-acylamlno-3-methyl-A 2-cephem-4-carboxylic acid and allows the activation of 3-methyl in the following reactions.

The production of the 4r-carbamoyl group in the compounds with the structure 8 is carried out according to similar methods to those described for the first synthesis sequence.

In a similar way as for the first sequence, the compounds 8 are also transferred to tetrazoles with the structure 9 according to methods which include the formation of the imino chloride and its reaction with the azide.

The removal of the "tetrazole-protecting" group in the compounds S > leading to 10 is carried out in the same manner as discussed

earlier when it comes to the first synthesis path and consists in bringing 9 into contact with trifluoroacetic acid/anisole at 30-50°c for a few hours.

Before the bromination of the activated 3-methyl substituent, the tetrazole 10 is blocked by alkylation with chloromethyl methyl ether. As the person skilled in the art will understand, the alkylation can take place, and does take place, at both the N^ and Nj positions, the alkylation being mainly at N2. Since the blocking group is removed in a

later steps in the sequence, it is practical that the two isomers are not separated after the alkylation. Both isomers also serve the same purpose and are equally useful.

The 3-methyl substituent in 11 is brominated with N-bromosuccinimide

in the presence of a peroxide, a known bromination method. After the completion of the bromination, the 3-bromoethyl compound is not isolated, but is reacted with a special nucleophilic reagent such as mercaptan or an acetate salt which gives 12.

The removal of the "amino-protecting" group in 12 is carried out according to methods discussed earlier in the first synthesis sequence0

In a similar way, the acylation of 13 is carried out by using the previously known methods, protecting or masking any functional groups in the acid to be acylated and which may compete during the acylation with the 7-amino group in A<?* ->c@five (13).

2 3 The reisomerization of the A -bond of 14 to the A -position is achieved by oxidation of the sulfur atom in the cephem molecule with a peracid such as m-chlorobenzoic acid ©, and gives 15. The treatment of 15 with stannous chloride and acetyl chloride results in the formation of the desired A<3 ->cephem with the structure 16.

The N^ and N2 blocking groups are removed from 16 by

use trifluoroacetic acid/anisole as indicated previously and<w>masking" groups are removed from the acyl group in the 7-acylamino group if these are present.

A modification of the second reaction sequence allows the preparation of compounds where R 1 is derived from phthalidyl or an alkanoyloxymethyl or 1-(alkanoyloxy)ethyl group. The procedure leading to the preparation of these compounds employs alkylation of the tetrazole group in 10 with a suitable alkanoyloxymethyl halide or 1-(alkanoyloxy)ethyl halide (Ulich, et al,

J. Am. Chem. Soc, 43, 660 (1921) and Éuranto, et al. Acta Chem. Scand., 20, 1273 (1966) or phthalidyl halide instead of chloromethyl methyl ether. Phthalidyl-, alkanoyloxymethyl- and 1-(alkanoyloxy)ethyl-substituted tetrazoles of the end products are precursors to the end products and even if these have no effect or have a relatively low effect in and of themselves against microorganisms, these are metabolized to the free tetrazole (R- ^H) when

they are injected parenterally into animals and humans. The speed of the metabolic transfer of these compounds to the free tetrazole

is such that you get an effective and prolonged concentration of the free tetrazole in the body. Such compounds are thus depot sources for the free tetrazole antibacterial agent.

As far as the antibacterial action of these precursor forms is concerned, both N 1 - and N 2 -substituted isomers are effective and useful.

". If phthalidyl-, alkanoyloxymethyl- or 1-(alkanoyloxy)-ethyl-, N^/N^-substituted tetrazoles are synthesized according to the second synthesis route, the reaction step (16-+6) is omitted, and only the masking groups are removed from the acyl part of the side chain.

There are alternative methods for the synthesis of the antibacterial products which contain a phthalidyl, alkanoyloxymethyl or 1-(alkanoyloxy)ethyl group in the Nj/N2 position in the tetrazole group. One method involves the alkylation of the base salt of a suitable 7-amino-3-substituted-4-(tetrazol-S-yl)-A<3->cephem, followed by the acylation of the 7-amino group as previously discussed.

The second method uses alkylation of the base salt of a suitable 7-acylamino-3-substituted-4-(tetrazol-5-yl)-A3-cephem» both methods include, if necessary, removal of "masked" groups from the side chain.

As for the second series of reactions leading to intermediates and final products according to the present invention

the preferred "amino-blocking" groups are the 2,2,2-trihalo-ethoxycarbonyl group.

As previously stated, it is a characteristic feature of them

acidic compounds according to the present invention, those where R^

or R^ is H or Q is carboxy, that they have the oven to form basic salts. The acid classes according to the present invention are transferred to the base salts by reacting the acid with a suitable base in an aqueous or non-aqueous medium. Such basic reagents used for the preparation of the salts can be of different types, and are intended to include such bases as organic amines, ammonia, alkali metal hydroxides, carbonates, bicarbonates, hydrides and alkoxides, as well as alkaline earth metal hydroxides, hydrides, alkoxides and carbonates. Examples of such bases are ammonia, primary amines such as n-propylamine, n-butylamine, aniline, cyclohexylamine, benzylamine, p-toluidln, ethylamine, octylamine, secondary amines such as dlcyclohexylamine and tertiary amines such as diethylaniline, N- methylpyrrolidine, N-methylmorpholine and 1,5-diaea-bicyclo[4.3.0]-5-nonene, sodium hydroxide, potassium hydroxide, ammonium hydroxide, sodium ethoxide, potassium methoxide, magnesium hydroxide, calcium hydride and barium hydroxide.

The person skilled in the art will easily understand that some of the compounds according to the present invention are sufficiently basic, namely the end products where Q©r amino, to form acid addition salts" these salts, especially pharmaceutically acceptable acid addition salts, are also covered by the present invention.

The usable intermediates according to the present invention which contain a free 7-amino group or a free tetrazole group (R^* R^aH) are also capable of forming acid addition salts and base salts. These salts are either used to characterize these intermediates, such as acid addition salts, or they are used in reactions, such as alkylation of the base salt of tetrazoles.

When utilizing the chemotherapeutic effect of these compounds according to the present invention which form basic salts, it is of course preferred to use pharmaceutically acceptable salts. Although insolubility in water, high toxicity, or lack of crystalline property may render some salts unsuitable or less suitable for use as such in a given pharmaceutical application, the water-insoluble or toxic salts may be converted to the corresponding acids by cleavage of the salts, or the can alternatively be transferred to desired pharmaceutically acceptable basic salts. The preferred pharmaceutically acceptable salts include sodium, aluminum, potassium, calcium, magnesium, ammonium and substituted ammonium salts, e.g. procaine, dibenzylamine, N,N-bis(dehydroabiethyl)ethylenediamine, 1-ephenamine, N-ethylpiperidine, N-benzyl-B-phenethylamine, N,N*-dibenzylethylenediamine, triethylamine, as well as salts with other amines which have been used for to prepare salts with the cefem compounds.

The new cephem compounds exhibit in vitro activity against a number of microorganisms, including both gram-positive and gram-negative bacteria. The beneficial effect can be easily demonstrated

by in vitro tests against various organisms 1 a brain-heart infusion medium by the usual double serial dilution technique.

The in vitro activity of the indicated compounds makes them useful for topical applications in the form of ointments, taps, etc., or for sterilization purposes, e.g. hospital equipment.

The new cefem compounds are effective antibacterial agents in vivo in animals, and include humans, not only by the parenteral route of administration, but also by the oral route of administration. The doctor will of course determine the optimal dose that will be most suitable for a given person, and this will vary with age, weight and the particular patient's response as well as the nature and

the degree of the symptoms, the type of bacterial infection to be treated and the pharmacodynamic properties of the particular

agent administered. It is often found that when the preparation is administered orally, larger amounts of it are required

active ingredient to give the same level as obtained with a smaller amount administered parenterally.

On the basis of what has been said above, an effective daily oral dose of the compounds according to the present invention in humans will be approx. 50-1000 mg/kg per day, with a preferred area from approx. 250-750 mg/kg per day in single or divided doses, and a parenteral dose from 25-500 mg/kg per day, with a preferred area from approx. 125-400 mg/kg per day which will effectively relieve the symptoms of infection. These values are only given to illustrate and they can of course be higher or lower in the various cases.

Preferred compounds according to the present invention

which are useful as intermediates are 7-(2',2•,2•-trichloroethoxycarboxamido)-3-methyl-4-[N-(p-methoxybenzyl)carbaroyl]-A3-cephem, 7-(benzyloxycarboxamido)-3- methyl- -[N-(p-methoxybenzyl)-carbamoyl]-A<3->cephem, 7-(N-triphenylmethylamino)-3-methyl-4-[N-(p-methoxybenzyl)carbamoyl]-A<3 ->cephem, 7-(2',2',2•-trichloroethoxycarboxamide)-3-acetoxymethyl-4-[N-(p-methoxybenzyl)carbamoyl]-A -cephem, 7-(benzyloxycarboxamido)-3- acetoxyethyl-4-[N-(p-methoxybenzyl)-carbamoyl]-^<3->cephem, 7-(N-triphenylmethylamino)-3-acetoxymethyl-4-[N-(p-methoxybenzyl)carbamoyl]-A3- cephem, 7-(2<1>,2•,2'-trichloroethoxycarboxamido)-3-raethyl-4- [N-(p-raethoxybenzyl)carbamoyl]-α2-cephem, 7-(2',2',2 '-trichloroethoxycarboxamido)-3-methyl-4-[l-(p-methoxybenzyl)tetrazol-5-yl]-α 3-cephem, 7-(N-triphenylamino)-3-methyl-4-thi-(p- methoxybenzyl)tetrazol-5-yl]-A -cephem, 7-amino-3-methyl-4-[1-(p-methoxybenzyl)tetrazol-5-yl]-A -cephem, 7-(benzyloxy-carboxamido) - 3-acetoxymethyl- 4-[1-(p-methoxybenzyl)tetrazol-5-yl]-A<3->cephem, 7-(2• ,2•,2'-trichloroethoxycarboxamido)-3-acetoxymethyl-4-[1-(p-methoxybenzyl)tetrazol-5-yl]-A -cephem, 7-amino-3-acetoxymethyl-4-[l-(p -methoxybenzyl)tetrazol-5-yl]-α<3->cephem, 7-(benzyloxycarboxamido)-3-(1-methyltetrazol-5-ylthiomethyl)-4-[1-(p-raethoxybenzyl)tetrazol-5-yl ]-A3-cephem, 7-(2',2',2'-trichloroethoxycarboxamido)-3-(2-methyl-1,3,4-thiadiazol-5-ylthiomethyl)-4-[1-(p-methoxybenzyl )tetrazol-5-yl]-A - i -cephem, 7-amino-3-(2-methyl-1,3,4-thiadiazol-5-ylthiomethyl)-4-[1-(p-methoxybenzyl)tetrazol- 5-yl]-A3-cephem, 7-(2',2',2'-trichloroethoxycarboxamido)-3-methyl-4-[1-(p-methoxybenzyl)tetrazol-5-yl]-A -cephem, 7 -(benzyloxy-carboxamido )-3-(2-methyl-1,3,4-thiadiazol-5-ylthiomethyl)-4-[ 1-(p-methoxybenayl)tetrazol-5-yl]-A<3->cephem , 7-amino-3-methyl-4-(tetrazol-5-yl)-A 3 -cephem, 7-amino-3-acetoxymethyl-4-(tetrazol-5-yl)-α<3->cephem, 7 -amino-3-(2-methyl-1,3,4-thiadiazol-5-ylthiomethyl)-4-(tefcrazol-5-yl)-α<3->cephem, 7-amino-3-(l-methyltetrazol-5 -ylthiomethyl)-4-(tetrazol-5-yl)-a 3-cephem, 7-(2',2',2'-trichloroethoxy ycarboxamido)-3-methyl-4-(tetrazol-5-yl)-A 2-cephem, 7-(2',2',2'-trichloroethoxycarboxamido)-3-methyl-4-[1-(methoxymethyl)tetrazole -5-yl]-A 2-cephem, 7-(2',2',2'-trichloroethokaycarboxamido)-3-methyl-4-[2-(methoxymethyl)tetrazol-5-yl]-A 2 -cephem, 7-amino-3-methyl-4-[1-(methoxymethyl)tetrazol-5-yl3-A -cephem, 7-amino-3-methyl-4-[2-(methoxymethyl)tetrazol-5-yl]-A <2->cephem, 7-amino-3-acetoxymethyl-4-t1-(methoxymethyl)tetrazol-5-yl]-A<2>-cephem, 7-amino-3-acetoxymethyl-4-[l-(methoxymethyl )tetrazol-5-yl]-A<2->cephem,

7-amino-3-(1-raethyltetrazol-5-ylthiomethyl)-4-[1-(methoxymethyl)-tetrazol-5-yl]-A 2-cephem, 7-amino-3-(1-methyltetrazol-5- ylthiomethyl)-4-[2-(methoxymethyl)tetrazol-5-yl]-A<2->cephem, 7-amino-3-(2-methyl-1,3,4-thiadiazol-5-ylthiomethyl)-4 -[1-methoxymethyl)tetrazol-5-yl]-2

A -cephem, 7-amino-3-(2-methyl-1,3,4-thiadiazol-5-ylthiomethyl)-4-[2-(methoxymethyl)tetrazol-5-yl]-A<2->cephem, 7-(2',2"^'-trichloroethoxycarboxamido)-3-bromomethyl-4-[1-methoxymethyl)tetrazol-5-yl]-

2

A -cephem and 7-(2',2*,2'-trichloroethoxycarboxamido)-3-bromo-methyl-4-[2-(methoxymethyl)tetrazol-5-yl]-A 2-cephem.

The preferred antibacterial agents according to the present invention are 7-phenylacetamido-3-methyl-4-(tetrazol-5-yl)-A -cephem, 7-phenoxyacetamido-3-methyl-4-(tetrazol-5-yl)-A 3 -cephem, 7-(2-thienylacetamido)-3-methyl-4-(tetrazol-5-yl)-A<3>-cephem, 7-phenylacetamido-3-methyl-4-[2-(pivaloyloxymethyl)tetrazol- 5-yl]-A -cephem, 7-phenoxyacetamido-3-methyl-4-[2-(pivaloyloxymethyl)tetrazol-5-yl]-A<3->cephem and 7-(2-thienylacetamido)-3-methyl -4-[2-(pivaloyloxymethyl)-tetrazol-5-yl J-A -cephem.

Antimicrobial data for a number of compounds according to the present invention against Streptomyces pyogenes are given in the following table. These experiments were carried out under standardized conditions where the nutrient broth containing different concentrations of experimental material was inoculated with the indicated organism, and the minimum concentration (MIC) at which no growth of the microorganism was obtained was determined.

The new antibacterial agents according to the present invention are remarkably effective for the treatment of a variety of infections caused by gram-negative and gram-positive bacteria in poultry and animals, and also humans. For such purposes, pure materials or mixtures thereof with other antibiotics can be used. They can be administered alone or in combination with a pharmaceutical carrier based on the chosen route of administration and standard pharmaceutical practice. They can e.g. administered orally 1 form of tablets containing such excipients as starch, milk sugar, certain types of clay etc, or only in capsules or in mixtures with the same or equivalent excipients. They can also be administered orally in the form of elixirs or oral suspensions which may contain flavoring substances, or coloring substances, or they can be injected parenterally, i.e. intramuscularly or subcutaneously. For parenteral administration, they are best used in the form of a sterile aqueous solution which can either be aqueous such as water, isotonic saline solution, isotonic dextrose, Ringer's solution, or non-aqueous, such as fatty oils of vegetable origin (cottonseed oil, peanut oil, corn oil, sesame oil) and other non-aqueous solvents that do not interfere with the therapeutic effect during preparation and are not toxic in the volume or quantity used (glyoerol, propylene glycol, sorbitol). In addition, preparations suitable for the preparation of preparations to order can be prepared from solutions prior to administration. Such preparations may comprise liquid diluents, e.g. propylene glycol, diethyl carbonate, glycerol, sorbitol, etc., buffering agents, as well as local anesthetics and inorganic salts to provide the desired pharmacological properties.

The following examples are given to illustrate the invention. Infrared (IR) spectra are measured as potassium bromide disks (KBr disks) or as Nujol mulls, and diagnostic absorption bands are indicated by 1 wavelength number (cm~^). Nuclear magnetic resonance spectra (NMR) have been measured at 60 MHz for solutions in deuterochloroform (CDCl3), perdeuterodimethylsulfoxide (DMSO-dg) or deuterium oxide (D2O), and the peaks are expressed in parts per million (ppm) "downfield a from tetramethylsilane or sodium 2,2-dimethyl-2-silapentane-5-sulfonate. The following abbreviations for the shapes of the peaks are used"

s, singlet, d, doublet, t, triplet, q, quartet, m, multiplet.

Example 1

7-(2•,2',2'-trichloroethoxycarboxamido)-3-methyl-4-[N-(p-methoxybenzyl)carbamoyl3~ A<3>- cephem

A. 7-(2',2°, 2°-trichloroethoxycarboxamido)-3-methyl 1-A 3-cephem^-carboxylic acid

To a suspension of 553 g of 7-amino-3-methyl~A3-cephem~4-carboxylic acid in 16.65 1 solution of 1:2 acetone/water, 600 g of sodium bicarbonate is added and the mixture is stirred at room temperature for 30 minutes. To the resulting solution, 600 g of 2,2,2-trichlorotoxycarbonyl chloride is added over 45 minutes and the reaction mixture is stirred for a further 18 hours. The mixture is extracted with ethyl chloride and the aqueous layer is separated and acidified to pH 2 with 5W hydrochloric acid. The acidified aqueous layer is extracted with fresh methylene chloride (3 sc 1.51 and 2 x 500 ml) and the combined organic layers are washed sequentially with 5N hydrochloric acid (2 x 500 ml) and water (3 x 500 ml) and dried over magnesium sulfate . The thick oil, which remains after the solvent has been removed in vacuo, is dissolved in 12 1 diethyl ether and added dropwise to 1.5 1 petroleum ether. The intermediate crystallizes slowly from the solution (70% yield).

NMR (DMSOdg)s 6 » 5.4 (q) 1H> 5.1 (d) 1H? 4.85 (s) 1H?3.42 (s) 2H, 2.0 (a) 3H. ,

IR (KBr disc) y max: 1770 cm""<*>(3-lactam carbonyl).

The product is stored in dioxane solution for use in the following reactions without further purification.

B. 7-(2',2•,2•-trichloroethoxycarboxamido)-3-methyl-4-[N-(p-ethoxy-benzyl)-carbamoyl3-α<3>-cephem.

A solution containing 1.34 g of 2,4-dinitrophenol in 35 ml of methylene chloride is treated with 4 g of 7-(2',2',2'-trichloroethoxycarboxamido)-3-methyl-A<3->cephem-4-carboxylic acid in the smallest possible amount of dioxane, followed by a solution of 2.1 g of dicyclohexylcarbodiimide in 20 ml of methylene chloride. After stirring the reaction mixture for 30 minutes at room temperature, dicycloxylurea is filtered, washed with methylene chloride, and the filtrate and washing solutions are combined and treated with 1.4 g of p-methoxybenzylamine in 14 ml of methylene chloride.

After 30 minutes at room temperature, the reaction mixture is washed with saturated sodium carbonate solution and the organic phase is dried over magnesium sulfate. The thick oil, which remains after the solvent has been removed under reduced pressure, is triturated with petroleum ether. The crystallized product is filtered and dried to give 4.0 g of a slightly yellow solid.

NMR (DMSOdg)s 6 => 8.5 (t) 1H, 7.0 (q) 4H, 5.3 (q) and

4.95 (d) 2H, 4.8 (s) 2H» 4.3 (d) 2H, 3.7 (s) 3H, 3.45 (broad) 2H

and 2 (s) 3Ho

IR (KBr disc) y: 1770 cm""<1>(3-lactam carbonyl).

Example 2

7-(2<8>,2',2'-trichloroethoxycarboxamido)-3-methyl-4-11-p-methoxy-

3

benzyltetrazol-5-yl-A - °cephem

To a suspension of 100 g of 7-(2',2",2'-trichloroethoxy-tarboxamido)-3-methyl-4-[N-(p-methoxybenzyl)carbamoyl]-A 3-cephem 12 1 chloroform is added dropwise 250 ml of toluene containing

30 g of phosgene within 15 minutes. After a further 15 minutes, a solution of 24 g of dry pyridine in 100 ml of chloroform is added dropwise over the course of 45 minutes, during which time a solution is obtained and carbon dioxide slowly evolves. The solution is stirred for 90 minutes and then concentrated to half the volume in vacuo to remove excess phosgene. Chloroform (2 1) is added

and 45 g of tetramethylguanidinium azide in 250 ml of chloroform are then added to the stirred solution. After 2 hours, the chloroform solution is washed successively with water (2 x 500 ml), saturated sodium carbonate solution (2 x 300 ml), 2.5E3 hydrochloric acid and water (1 x 250 ml). The organic phase is dried over magnesium sulphate and concentrated

in vacuo to a gummy solid, 102 g. The product is further purified by chromatography on a column of alumina (306 g) using chloroform as solvent (408 ml) and eluent (1.5 L). The eluate is added with stirring to 6 1 petroleum ether, and this results in precipitation of the product as a light brown solid, 79 g.

NMR (CDCl3)s 6 ■ 6.9 (q) 4Hr 5.4 (m) 3H, 4.9 (d) 1H,

4.7 (s) 2H, 3.7 (s) 3H, 3.3 (q) 3H and 1.4 (s) 3H.

IR (KBr disc) y: 1770 cm"<*1>(fl-lactam carbonyl) „

Similarly, 250 mg of 7-(2',2•,2•-trichloroethoxycarboxamido)-3-methyl-[N-(p-methoxybenzyl)carbamoyl]-A<3->cephem, 0.007 me pyridine and 2.6 mg of phosphorus pentachloride in 6 ml of chloroform the corresponding imino chloride which, on treatment with tetramethylguanidinium azide (lg) in 12 ml of chloroform, gives 220 mg of the desired tetrazole, which is in all respects identical to that obtained above.

Example 3

7~amino-3-methyl-"4-[1-(p-methoxybenzyl)tetrazol-5-yl]-A-cephem

25 g of zinc dust, and the mixture is stirred. vigorously for 3 hours at room temperature. The mixture is filtered and the filtrate is added to a large volume of water and extracted with chloroform.' The organic layer is washed with water and the product is extracted from the organic phase with IN hydrochloric acid. The aqueous phase is washed with chloroform, basified with 2N sodium hydroxide solution to pH 7 and extracted with chloroform. The chloroform layer is dried over magnesium sulfate and concentrated under reduced pressure to a small volume. When the concentrate is added to petroleum ether, the product precipitates. The solid is filtered off and dried, 30 g.

NMR (DMSOdg): 6 « 7(2q) 8H, 5.5(s) 2H, 5.4 (d) 1H, 5.05 (d) 1H, 3.7 (s) 3H, 3.6 ( s) 2H, 2.2 (s) 3H and 1.4 (s) 3H»

Example 4

7- amino- 3- methyl- 4-( tetrazol- 5- yl)- A 3- cephem

A mixture of 7.0 g of 7-amino-3-methyl-4-(1-[p-methoxybenzyl]-tetrazol-5-yl)-A<3->cephem in 50 ml of trifluoroacetic acid/anisole (4sl)

allowed to stand at 50°C for 5 hours. The solution is then poured

into a large volume of stirred diethyl ether and the precipitate that forms is filtered off. The solid is dissolved in water, which is extracted

several times with ethyl acetate, adjusted to pH 7.0 with 2N aqueous sodium hydroxide and extracted again. The aqueous solution containing the product is freeze-dried to give 3.2 g of the desired compound (which contains some sodium trifluoroacetate).

NMR (DMSOdg)s 6 =» 5.0 (d) 1H, 4.6 (d) 1H, 3.4 (q) 2H,

1.9 (s) 3H and 1.3 (s) 3H.

Example 5

3 7-( benzyloxycarboxamido)- 3- methyl- A - cephem- 4~ carboxylic acid

Sodium bicarbonate (500 g) is added in portions during

3

45 minutes to a stirred suspension of 7-amino-3-methyl-A-cephem-4-carboxylic acid (560 g) in cold water (5.6 liters) and acetone (5.6 liters). When the effervescence has ceased, benzyl chloroformate (490 g) is added and the solution is stirred at room temperature overnight. The reaction mixture is then extracted with ethyl acetate (2x1 liter), and the pH of the separated aqueous layer is adjusted to 2 by adding 2N hydrochloric acid. This mixture is then extracted with ethyl acetate

(2x1 liter) and the solvent from the separated organic layer is removed in vacuo. The resulting solid is dissolved in hot methanol (2 L) and diluted with hot water (5 L) to give 7-(benzyloxycarboxamido)-3-methyl-A 3-cephem-4-carboxylic acid as a white solid ( 750 g), which is dried in vacuum at 60°C.

NMR (DMSOdg): δ = 8.3 (d) 1H, 7.3 (m) 5H, 5.4 (g) 1H,

5.1 (d) 1H, 5 (s) 2H, 3.4 (q) 2H and 2.1 (s) 3H,

Example 6

7-(Benzyloxycarboxamido)-3-methyl-4-[N-(p-methoxybenzyl)-carbamoyl3- A 3- cephem

A mixture of 7-(benzyloxycarboxamido)-3-methyl-A<3->cephem-4-carboxylic acid (34.8 g) and 2,4-dinitrophenol (18.4 g)

dissolve 1 dry dioxane (700 ml). Dicyclohexylcarbodiimide (20.6 g) is then added and the solution is stirred for 20 minutes at room temperature. The precipitated dicyclohexylurea is then filtered off, and the filtrate is treated with p-methoxybenzylamine (13.7 g) which is added 1 over 10 minutes. After a further 15 minutes, the reaction mixture is diluted with dry ether (1.4 litres).

The precipitate is filtered off, washed with dry ether (500 ml), and

dried in vacuum at 50°C, and 40 g of the product is obtained as a white,

solid.

NMR (DMSOdg): 6 =8.4(m) 2H, 7 (q) 4H, 5.3 (q) 1H,

5.1 (s) 2H, 5 (d) 1H, 4.3 (d) 2H, 3.7 (s) 3H, 3.3 (s) 2H and 2 (s) 3H.

Example 7

7- amino- 3- methyl- 4-[ N-( p- methoxybenzyl) carbamoyl 3- A^ r-cafem

7-(Benzyloxycarboxamido)-3-methyl-4-[N-(p-methoxybenzyl)-carbamoylQ-A 3-cephem (20 g) is dissolved in hydrobromic acid/acetic acid (200 ml) and the solution is stirred at room temperature until gas evolution ceases . The mixture is poured into a large volume (about 2 liters) of stirred ether and the resulting skimming is collected. The solid is dissolved in water and the aqueous solution is washed with ethyl acetate (2 times)* The aqueous solution is adjusted to

pH 7.5 with sodium bicarbonate and the suspension is extracted with chloroform. The chloroform extract is dried (MgSO4) and evaporated under reduced pressure, and the product is obtained as a pale yellow solid (7.7 g).

NMR (DMSOdg)16 - 8.0 (t) 1H, 7.0 (q) 4H, 4.8 (d) 1H,

4.6 (d) IK, 4.4 (d) 2H, 3.7 (s) 3H, 3.2 (q) 2H, 2.1 (s) 2H and 2.0 (s) 3H<

Example 8

7-(N-triphenylmethylamino)-3-methyl-4-[N-(p-methoxybenzyl)carbamoyl]-A3-cephem To 24 g 7-amino-3-methyl-4~[N-(p-methoxybenzyl) carbamoyl]-A 3-cephem in chloroform (ethanol-free, 300 ml) is added triphenylmethyl chloride (21.0 g) and triethylamine (7.64 g). The mixture stands at room temperature under the exclusion of light for 15 hours. The solution is then diluted to multiple volumes with chloroform and washed twice with water. The organic solution is dried (MgSO^) and

evaporates to dryness, and you get the product as a foam"

Trituration with light petroleum ether gives a pale yellow solid (40 g).

NMR (CDCl3)s 6«7.2, 9H, (q) 1H, 4.4 (d) 2H, 4.1 (d) 1H, 3.7 (s) 3H, 3.0 3H and 2.0 (s) 3H.

Example 9

7-(N-triphenylmethylamino)-3-methyl-4-[1-(p-methoxybenzyl)-tetrazol-5-yl]- A<3>- cephem

7-(N-triphenylmethylamino)-3-methyl-4-[N-(p-methoxybenzyl)-carbamoyl]-A 3-cephem (2.8 g) in ethanol-free chloroform (7.5 mL) containing pyridine (0 .6 g) is treated with phosgene in chloroform (3.7 ml, 1.7M). The reaction mixture is stirred for 30 minutes at room temperature. Excess phosgene is removed under reduced pressure and further chloroform is added until the volume of the solution is 10 ral. Tetramethylguanidine hydrogenazide (2.4 g) in chloroform (5 ml) is added to the solution prepared above and the reaction mixture is stirred at room temperature for 20 minutes. The solution is washed with water, dried (MgSO^) and evaporated under reduced pressure, and an oil is obtained. Trituration with light petroleum ether gives 2.2 g of the product.

NMR (CDCl 3 ): .6 =» 7.2 19 H, 5.3 (s) 2 H, 4.4 (q) 1 H, 4.2 (d) 1 H, 3.6 (3) 3 H, 3.0 3H and 1.2 (s) 3H.

Example 10

7-amino-3-methyl-4-[l-(p-methoxybenzyl)tetrazol-5-yl]- A 3-cephem 7-(N-triphenylmethylamino)-3-methyl-4-[l-(p-methoxyb ©nzyl)-tetrazol-5-yl]-A 3-cephem (1.5 g) is treated with formic acid (30 ml) and the solution is allowed to stand at room temperature for 30 minutes* The solution is poured into water (100 ml) and the aqueous the suspension is washed with ethyl acetate. The aqueous solution is adjusted to pH 7.5 with 2N aqueous sodium hydroxide and the mixture is extracted with chloroform. The organic solution is dried (MgSO^) and evaporated under reduced

pressure, and you get a foam. Trituration with light petroleum ether gives 400 mg of the product as a solid.

NMR (CDCl3)i 6 » 7.0 (q) 4H, 5.4 (s) 2H, 4.9 (d) 1H, 4.6 (d) 1H, 3.7 (s) 3H, 3.3 (q) 2H, 2.0 (s) 2H and 1.4 (s) 3H.

Example 11 7-[D~(a-amino-g-phenyl)acetamido]~ 3-methyl-4-(tetrazol-5-yl)-A3-cephem

A. 7~[D-α-t-butoxycarboxainido-o-phenyl)acetamido]-3-*methyl-4-(tetrazol-5-yl)- A 3-cephem

D-(α-t-butoxycarboxyldo-H-phenyl)acetic acid (0.251 g) in tetrahydrofuran (2 ml) is treated with triethylamine (0.101 g) and

ethyl chloroformate (0.108 g) at -10°C. After 15 minutes at this temperature, the sodium salt of 7-amino-3-methyl-4-(tetrazol-5-yl)-A-cephem (0.400 g) in water (3 ml) is added with stirring in one portion. The mixture is diluted with tetrahydrofuran (2 ml) and allowed to warm to room temperature. After 1 hour, the solution is adjusted to pH 2 with 2N hydrochloric acid and the suspension is extracted with ethyl acetate. The ethyl acetate solution is washed with water, dried (MgSO 4 ), concentrated under reduced pressure and poured into a large volume of light petroleum ether, and a white solid is obtained. The compound is recrystallized from chloroform (316 mL).

NMR (DMSOdg): 6 » 7.6 (d) 1H, 7.2 5H, 6.0 (d) 1H, 5.6 (q) 1H, 5.2 (d) 1H, 4.9 (d) 1H, 3.3 (q) 2H, »,0 (s) 3H.bg 1.2 (s) 9H.

B. 7-[D-(α-ajnino-q-phenyl)acetamido]-3-methyl-4-(tetrazol-5ryl)-A

Cepheme trifluoroacetate 7-[D-(a-t-butoxycarboxamido-o-phenyl)acetamido]-3-methyl-4-(tetrazol-i-yl)-A<3->cepheme (26 mg) is dissolved in trifluoroacetic acid

(0.5 ml) and allowed to stand at room temperature for 20 minutes.

Ether is added to the solution and the precipitate is collected (24 mg).

NMR (DMSOd 6 -D 2 O) t 6 * 7.4(s) 5B, 5.6(d) 1H, 5.1(d) 1H, 5.0(s) 1H, 3.4, 2H and 2.0 (s) 3H.

Example 12 7-[D-o-amino-α-(p-hydroxyphenyl)acetamido]-3-methyl-4-(tetrazol-5-yl)-α3-cephem

The procedure in example 11 Ac?and B is repeated by starting from the necessary D-[o-t-butoxycarboxamido-a-(p-hydroxyphenyl)]-acetic acid instead of D-(a-t-butoxycarboxamido-o-phenyl)acetic acid, and you get the product.

IR (CH2C<1>2)<:><Y>najt8« 1780 cm"<1>(p-lactam-carbonyl)" 1660 cm"1

(-C0NH-).

Example 13 7-(D-α-hydroxyphenylacetamido)-3-methyl-4-(tetrazol-5-yl)-α3-cephem A. 7-(D-o-formyloxyphenylacetamido)-3-methyl-4-(tetrazol-5- yl)-A -

cephem

D-O-formylmandelic acid (1.8 g) in ether (15 ml) is treated with oxalyl chloride (2.5 ml) and a drop of dimethylformamide. After the vigorous reaction has subsided (30 min.), the ether is removed under reduced pressure and the residue in THF (10 ml) is added to a stirred solution of 7-amino-3-methyl-4-(tetrazol-5-yl)- A -cephem (1 g) in a mixture of water/THF (lii, 20 ml) containing an excess of sodium bicarbonate. After stirring for a further 1 hour at room temperature, the mixture is adjusted to pH 2 with 2N hydrochloric acid and the mixture is extracted with ethyl acetate. The organic solution is washed with water, dried (MgSO4) and evaporated under reduced pressure, and the desired product is obtained as a solid (350 mg).

NMR (DMSOdg)» 6 - 8.2 (s) 1H, 7.2(s) 5H, 6.0(s) 1H, 5.5(q)

1H, 5.1(d) 1H, 3.5(s) 2H and 2.0, 3H.

B. 7-( D- g- hydroxyphenylacetamido)- 3- methyl- 4-( tetrazol- 5- yl)- A 3- cephem

7-(D-a-formyloxyphenylacetamldo)-3-methyl-4-(tetrazol-5-yl)-A3-cephem (0.34 g) is dissolved in an excess of aqueous sodium bicarbonate and allowed to stand at 30°C for 3 hours. The solution is adjusted to pH 2 with 2N hydrochloric acid and the suspension is extracted with ethyl acetate. The organic solution is dried (MgSO4) and evaporated under reduced pressure, and the product is obtained as a solid (0.26 g).

NMR (DMSOdg): 6 » 9.0, 1H, 7.3(s) 5H, 5.6(q) 1H, 5.2(d) 1H, 5.1(s) 1H, 3.6(8 ) 2H and 2.0(s) 3H.

Example 14

'3

7-( 2- cyanoacetamido)- 3- methyl- 4-( tetrazol- 5- yl)- A - cephem

To a stirred suspension of 7-amino-3-methyl-4-(tetrazol-5-yl)-α 3-cephemtosylate salt (576 mg) in methylene chloride (20 mL) under nitrogen was added triethylamine (450 mg). After stirring for 20 minutes at room temperature, this solution is treated with N-hydroxy-succinimide ester of cyanoacetic acid (470 mg), all in one portion. After stirring overnight under nitrogen, the reaction mixture is poured into 1 water (30 ml) and the pH of the aqueous phase is adjusted to 8.0. The methylene chloride layer is separated. The aqueous phase is acidified

pH 2 and then extracted with ethyl acetate. The ethyl acetate layer is washed with water, dried (sodium sulfate) and the solvent is removed

under vacuum* Trituration of the residue with dry ether gives the product as a faint yellow solid (55 mg)•

NMR (DMSOdg)i 6 » 5.7 (q) 1H, 5.2 (d) 1H, 3.8 (s) 2H,

3.6 (s) 2H and 2.0 (d) 6H.

Example 15 7-(2-Carboxyphenylacetamido)-3-ethyl-4-(tetrazol-5<-yl)-A3-cephem sodium salt

Phenylmalonic acid (0.79 g) is dissolved in distilled water (25 ml) and the pB of the solution is adjusted to 6.0 by adding 2N sodium hydroxide solution. A solution of 7-amino-3-methyl-4-(tetrazol-5-yl)-A<3>-cephem (0.65 g) in 10 ml of distilled water (pH adjusted to 6 by addition of 2N sodium hydroxide solution) is added then, and the reaction mixture is cooled to 0°C. 1<->ethyl-3-(3-dimethylaminoprop-1-yl)carbodiimide (1.6 g) is then added and the solution is stirred for 3.5 hours, and during this time the pH is kept in the range 6.1-6, 3 by the dropwise addition of dilute hydrochloric acid. The pH is then raised to 7.3 by adding saturated sodium bicarbonate solution, and the reaction mixture is extracted with ethyl acetate. The extract is discarded. The aqueous phase is then acidified to pH 3 by using

0.4M phosphoric acid, and extracted again with ethyl acetate (two 50 ml portions). The latter extract is dried and concentrated to a volume of approx. 25 ml. A solution of sodium 2-ethyl hexanoate (1 g) in 20 ml of ethyl acetate is then added to this solution. The precipitate that forms is filtered, and 0.75 g of the disodium salt of 7-(2-carboxy-2-phenylacetamido)-3-methyl-4-(tetrazol-5-yl)-A 3-cephem is obtained. The infrared spectrum of the product (KBr disc) shows absorptions at 1760 cm""<*>(3-lactam carbonyl), 1660 cm 1 (amide X band) and 1600 cm""<1>(carboxylate carbonyl) .

NMR (D 2 O)i 6 - 7.4 (m) 5H, 5.7 (d) 1H, 5.2 (d) 1H, 3.4(ra) 2H and 1.9(s) 3H.

Example 16

By using 2-thienylmalonic acid instead of phenylmalonic acid in method 1 example 15, 7-ta-(2-thienyl)-a-carboxyacetamido]-3-methyl-4-(tetrazol-5-yl)-<3-> cephem.

NMR (DMSOdg)t 6 - 2.1 (s) 3H, 3.3( ) 2H, 5(s) 1H, 5.6 (m) 1H, 6.8-7.5(m) 3H.

IR (KBr-ski<ve>)<Tm>ax1770cm""<1>(3-lactam-carbonyl),

1670 cm<**1>(-C0NH-).

Example 17

7- f enylacetamldo- 3- methyl~ 4- (tetrazol- 5- yl)- »A3- cef em

Phenoethyl chloride (2.3 g) in acetone (5 ml) is added dropwise to a stirred solution of 7-amino-3-methyl-4-(tetrazol-S-yl)-A<3->cephem sodium salt (1.31 g) in aqueous acetone (20 ml) for 20 minutes at room temperature. Sodium hydroxide (2N) is added at the same time to keep the pH at 8 ± 0.5. After the addition, the solution is stirred for a further 15 minutes. Acetone is removed under reduced pressure and the pH of the aqueous solution is adjusted to 2 with 2N hydrochloric acid. The suspension is extracted with ethyl acetate, the organic solution is dried (MgSO 4 ) and evaporated under reduced pressure, and an oil is obtained which solidifies after trituration with ether (440 mg).

NMR (DMSOdg)i6 - 9.0(d) 1H, 7.2(s) 5H, 5.6(q) 1H, 5.2(d) 1H, 3.5(a) 4H and 2.0( s) 3H.

Example 18

7- phenoxyacetamido- 3- methyl- 4-( tetrazol- 5- yl)- A3- cephem

Fenokay acetyl chloride (2.56 g) 1 acetone (5 ml) is added

dropwise to a stirred solution of 7-amino-3-raethyl-4-(tetrazol-5-yl)-A 3-cephem sodium salt (1.31 g) in aqueous acetone (30 mL) over 20 min at room temperature . Sodium hydroxide (2N) is added at the same time to keep the pH at 8 * 0.5. The solution is then stirred for a further 15 minutes. The acetone is removed under reduced pressure and the pH of the aqueous solution is adjusted to 2 with hydrochloric acid (2N). The suspension is extracted with ethyl acetate and the organic solution is dried (MgSO4)• Evaporation and trituration of the residue gives the product as a solid (370 mg).

NMR (DMSOdg)t 6 - 9.0<d) 1H, 7.0(m) 5H, 5.6(q) 1H, 5.2(d) 1H, 4.6(s) 2H, 3.8 (s) 2H and 2.0(s) 2H.

Example 19

By applying the procedure in examples 17 and 18 and starting from the required reagents, the following A 3 compound is obtained

7-bromoacetamido-3-methyl-4-(tetrazol-5-yl)-A<3->cephem,

NMR (DMSOdg)t 6 - 2.15(s) 3K, 3.7(m) 2H, 4.1(s) 2H, 5.4(d) 1H and 5.2(m) 1H,

7-(o-azido-o-phenylacetamido)-3-methyl-4-(tetrazol-5-yl)-A 3-cephem,

NMR (DMSOdg)s 6° 9.2(s) 1H, 7.35(s) 5H, 5.6 (s) 1H, 5.15(q) 2H, 3.58(s) 2H and 1.98 (a) 3H.

7-Co-(tetrazol-5-yl)acetamido]-3-methyl-4-(tetrazol-5-yl)-A<3->cephem,

NMR (DMSOdg)j 6 - 2.0(s) 3H, 3.6(s) 2H, 5.05-5.15(d) IR, 5.15(3) 2H, 5.5-5.8 (q) 1H and 9.5(a) 1H,

7-(α-phenylthioacetamido)-3-ethyl~4-(tetrazol-5-yl)-α<3->ce£em,

NMR (DMSOdg)i 6 - 9.18(d) LR, 7.23(ra) 5H, 5.63(q) LH, 5.35(d) 1H, 3.77(8) 2B, 3.6 (m) 2B and 2.03(s) 3R,

7-[o-(2,6-dimethoxyphenyl)carboxamido]~3-ethyl-4-(tetrazol-5-yl)-

3

A-cephem,

NMR (DMSOdg) t 6 - 2.0(s) 3H, 3.6(a) 2H, 3.7 (s) 6H, 5.05-5.15(d) LH, 5.5-5.8 (q) 1B, 6.5-6.7(d) 2B and 7.0-7.3(ra) IB,

7-(2-thienyigLyoxyLamido)-3-met<y>L-4-(tetrazoL-5-yl)-A<3>-cephem,

NMR (DMSOdg)i 6 - 2.0(s) 3H, 3.6(s) 2H, 5.1-5.15(d) 1H, 5.4-5.7(q) 1H, 7.05 -7,15(m) IB and 7,8-8,1 (m) 2H, and 7-Co-(2-thienyl)acetamido]-3-methyl-4-(tetrazol-5-yl)-A 3 -cephem,

NMR (DMSOdg)t 6 « 2.0(s) 3H, 3.6(s) 2B, 3.8 (a) 2B, 5.1-5.15(d) 1H, 5.5-5.8 (q) 1H, 6.95-7.0(d) 2H and 7.1-7.2 (m) 1H.

Example 20

7-[α-hydroxy-c-(2-thienyl)acetamido]-3-methyl-4-(tetrazol-5-yl)-α<3->cephem

7-(2-thlenylglyoxylamido)-3-methyl-4-(tetrazol-5-yl)-α-cephem (1.50 g) in water (30 ml) is transferred to sodium urate with 2N sodium hydroxide. This solution is cooled in ice and anhydrous sodium acetate (1.53 g) is added. Sodium borohydride (276 mg) is added in small portions over 45 minutes. The pH is maintained at 8 by alternately adding glacial acetic acid and sodium borohydride. After the additions, the pH is kept at 8 for 45 minutes with the ice bath removed. The solution is layered with ethyl acetate and the pB is adjusted to 2 with 40% phosphoric acid. The organic layer is separated, washed with water, dried (Na 2 SO 4 ), filtered and concentrated in vacuo until the solid crystallizes. The product is filtered, washed with dry ether and dried under vacuum, 360 mg.

NMR (DMSOdg)t 6 7.2 (ra) 1H, 7.0, 2H, 5.4(m) 1H, 5.0(s) 2H, 3.5, 2H and 2.0(s) 3H.

Example 21 7-[2-(2-methyl-1,3,4-thiadiazol-5-yl-thio-acetamido)]-3-methyl-4-(tetrazol-5-yl)-A 3-cephem

Triethylamine (0.2 ml) is added to a cold, 0°C suspension

of 1-(2-bromoacetamido)-3-methyl-4-(tetrazol-5°yl)-A 3-cephem (0.5 g)

1 methylene chloride (15 ral). To the resulting slightly yellow solution is added a suspension of 2-methyl-1,3,4-thiadiazole-5-thiol (0.19 g) and the solution is stirred at 0°C. After 2 hours, the precipitated product is collected by filtration (0.3 g).

NMR (DMSOdg)a 6 - 9.3(d) 1H, 5.6(m) 1H, 5.2(d) 1H, 4.15(s) 2H, 3.6(s) 2H, 2.7 (s) 3H and 2.05(s) 3H.

Example 22

The procedure 1 example 21 is repeated, starting from 7-(2-broaacetamido)-3-methyl-4-(tetrazol-5-yl)-A 3-cephem and 1-m@tyl-5-mercaptotetrazole (Lieber, et al ., Can. J. Chem. 37 101 (1959)), and one gets

7-Ca-(1-methyltetrazol-5-ylthio)acetamido]-3-methyl-4-(tetrazol-5-yl)-zP-cephem.

NMR (DMSOdg)s 6 » 9.28(d) 1H, 5.6(q) 1H, 5.24(d) 1B, 4.13(s) 2H, 3.95(s) 3H, 3.59 (m) 2H and 2.01(©) 3H.

Example 23

The procedure 1 example 21 is repeated by using 4-mercapto-pyridine and 7-(2-bromoacetamido)-3-methyl-4-(tetrazol-5-yl)-A 3-cephem, you get 7-[2-<4 -pyridylthio)acetamido]-3-methyl-4-(tetrazol-5-yl)-A 3-cephem.

NMR (DMSOdg)86 « 9.32(d) 1H, 8.41(m) 2H, 7.33(m) 2H, 5.7-5.6(q) 1H, 5.25(d) 1H, 3.95(d) 2B, 3.6(m) 2H and 2.03(c) 3H.

Example 24

7-(2*,2',2'-trichloroethoxycarboxamido)-3-acetoxymethyl-4-[N-(p-methoxybenzyl)carbamoyl]- A 3- cephem

A. 7-(2•,2',2 1-trichloroethoxycarboxamido)-3-acetoxymethyl-A 3-cephem-4-carboxylic acid

Sodium bicarbonate (300 g) is added portionwise to a stirred suspension of 7-amino-cefaiosporaneyr© (408 g) in aqueous acetone (2.1, 5 11 tert 2.5 liters). 2,*, 2', 2'-trichloroethyl chloroformate (350 g) is then added dropwise over 45 minutes and the mixture is stirred for a further 5 hours at room temperature.

The acetone is removed in vacuo and the aqueous solution is then diluted to 10 liters with distilled water. The pH of the solution is adjusted to 2 with 50% hydrochloric acid, and the mixture is extracted with ethyl acetate (5 at 1.5 litres). The combined organic layers are washed with water (2x2 litres), dried (MgSO 4 ) and the solution is concentrated to a volume of 1.5 litres.

The concentrate is added slowly to petroleum ether (b.p. 60-80°C, 15 litres) and the precipitated solid material is washed with petroleum ether (b.p. 30-40°C, 2x2 litres). The separation is dried in vacuum at 45°C, and 7-(2•,2<1>,2•^trichloroethoxycarbonyl)-amino-3-acetoxymethyl-A 3-cephem-4-carboxylic acid is obtained as a white solid ( 490 g).

NMR (DMSO dg )t. 6 - I0(d) 1H, 5.5(q) 1H, 5.1(d) 1H, 4.8(q) 2H, 4.8(3) 2H, 3.5(s) 2H and 2, 0(s) 3H.

B. 7-(2',2•,2'-trichloroethoxycarboxamido)-3-acetoxy-4-EN-(p-methoxybanzyl)-carbamoyl]-A 3-cofora'

By starting from the acid in example 24A and following the procedure

in example IB, the desired product is obtained in a moderate yield.

NMR (CDCl 3 ) : 6"2, 0{ a) 3H, 3.4(s) 2H, 3.8(s) 3H, 4.45(d) 2H, 4.75(S) 2H, 4.9 (m) 3H, 5.5(q) 1H, 6.2(b) 1H and 7.1(q) 4B2.

Example 25

7-(2',2•,2•-trichloroethoxycarboxamido)-3-(2-methyl-1,3,4-thiadiazol-5-ylthiomethyl)- 4°[ N-( p -methoxybenzyl)carbamoyl3- A - cephem A. 7-(2<1>,2<9>,2'-trichloroethoxycarboxamido)-3-(2-methyl-1,3,4-thia-diazol-5-yl 1Qmethyl)- A 3- cephem- 4- carboxylic acid

By proceeding as indicated in Example 24A, but using the cephem derivative 7-amino-3-(2-methyl-1,3,4-thiadlazol~5-ylthiomethyl)-A-cephem-4-carboxylic acid (U.S. Pat. 3.641.021) you get the desired product.

NMR (CDCl3)s 6 • 5.4(m) 2H, 5.1(d) 1H, 4.85(s) 2H, 4.4(q) 2H, 3.7 (s) 2H and 2.7 (s) 3H.

B. 7-(2',2<8>,2•-trichloroethoxycarboxamido)-3-(2-methyl-1,3,4-thladiazol-5-ylthiomethyl)-4-[ N-( p -methoxybenzyl)carbamoyl ]- A 3- cefem By using the product from example 25A and applying the method indicated in examples 24B and IB, the desired intermediate product can be synthesized.

NMR (CDCl 3 ): 6 = 7.0(q) 2H, 6.1(m) 1H, 5.4(q) 1H, 4.9(d) 1H, 4.7(s) 2H, 4, 4 (m) 4H, 3.75(s) 3H and 3.6(s) 2H.

Example 26

7-(2',2',2'-trichloroethoxycarboxamido)-3-aoethoxymethyl-4-£1-(p-methoxybenzyl)tetrazol-5-yl 3-A-cephem

Fos forpantochloride (1.68 g, 8 mmol) is added to a stirred solution of 7-(2*,2*,2'-trichloroethoxycarylxamido)-3-acetoxymethyl-4-[N-(p-methoxybenzyl-9carbamoyl]-A 3-cephem (2.26 g, 4 mmol) and

pyridine (0.64 g, 8 mmol) in dry ethanol-free chloroform (30 mL)

at 0-5 CV After 30 minutes, the nuclear magnetic resonance spectrum indicates that the amide has been completely converted (the two proton doublets at 6 4.45 have disappeared). A solution of tetramethylguanldinlumazide (6.3 g, 40 mmol) in dry ethanol-free chloroform (20 ml) is added dropwise over 10 minutes to the cooled solution. After a further 10 minutes, the ice bath is removed and the reaction mixture is stirred at ambient temperature for 30 minutes. The reaction mixture is washed with water, aqueous sodium bicarbonate solution (3 x), 6N hydrochloric acid (3 x), water and dried over magnesium sulfate. Evaporation of the solvent gives impure product as a light brown solid (1.5 g)• Column chromatography on silica gel which initially uses hexane as eluent and gradually adds increasing amounts of otter, and finally elutes with 100% otter, gives the purified product (by thin-layer chromatography)•

NMR (DMSOdg) 6 » 1.8(8) 3H, 3.6(q) 2H, 3.9(s) 3H, 4.2(q) 2H, 4.8(s) 2H, 5.1( d) 1H, 5.6(m) 3H, 6.3(d) 1H and 7.4(q) 4H.

Example 27 7-(2',2•,2<1->trichloroethoxycarboxamido)-3-(2-methyl-1,3,4-thiadlazol-5-ylthiomethyl)-4-[1-( p -methoxybenzyl) tetrazole - 5- yl3- A •3- cephem

When starting from the cephem compound in example 25B and

following the procedure in example 26, the desired product is obtained.

NMR (CDCl 3 )s 6° 7.0(q) 4H, 5.5(s) 2H, 5.4(m) 2H, 4.7(s) 2H, 4.1(m) 2H, 5.75 (a) 3Hog5.65(m) 2H.

Example 28

7- amino- 3- aoethokaymethyl- 4-[ 1-( p- methoxybenzyl) tetrazol- 5- yl)- A3-befem

Activated zinc dust (300 mg) is added to a stirred solution of 7-(2',2',2*-trichloroethoxycarboxamido-3-acetoxymethyl-4-[1-(p-methoxybenzyl)tetrazol-5-yl]-A -cephem (300 mg, 0.5 mmol) in 90% acetic acid (3 mL) and the suspension is stirred at ambient temperature for 30 min. The reaction mixture is filtered and evaporated to dryness. The resulting yellow foam is dissolved in chloroform and the solution is extracted twice with dilute hydrochloric acid. The aqueous extracts are mixed with chloroform, and dilute sodium hydroxide is added to the stirred mixture to pH 7.0. The mixture is filtered to remove precipitated inorganic salts, the layers are separated, and the aqueous solution is extracted twice with chloroform. The combined organic extracts are washed with water and dried over magnesium sulfate Evaporation of the solvent gives

the product as a faint yellow solid (110 mg), which is purified

by thin-layer chromatography,

NMR (CDCl 3 ): 6 • 1.9(s) 1H, 2.0(s) 3H, 3/45(q) 2H, 3.8(s) 3H, 4.3(q) 2H, 3.8 (s) 3H,4,3(q) 2H, 4.65(d) 1H, 4.95(d) 1H, S,5(s) 2H and 7, 1(q) 4H*

In a similar way, starting from 7-(2',2',2<*->trichloroethoxycarboxamido)-3-(2-methyl-1,3,4-thiadlazol-5-ylthiomethyl)-4- [ 1-(p-methoxybenzyl)tetrazol-5-yl]-A-cephem and following the procedure in Example 28 above, 7-amino-3-(2-methyl-1,3,4-thiadiazol-5-ylthiomethyl)-4 -[1-(p-Methoxybenzyl)tetraz'ol-5-yl]-A 3-cephem.

NMR (CDCl3)i 6 « 7.0(q) 4H, 5.55(s) 2H, 4.9(d) 1H, 4.6(m) 1H, 3.8(s) 3H, 3.65 (m) 2H and 2.7(s) 3H.

Example 29

7-amino-3-(2-methyl-1,3,4-thiadiazol-5-ylthiomethyl)-4-tetrazol-5-yl)-' A3-cephem

7-amino-3-(2-methyl-1,3,4-thiadiazol-5-ylthiomethyl)-4-[1-(p-methoxybenzyl)totrazol-5-yl3-A 3-cephem (125 mg) is dissolved in acetone (2 ml) and added p-toluenesulfonic acid (60 mg) in acetone (1 ml).

The salt precipitates as a brown oil after adding ether to the mixture. The solvent is decanted and the residue is washed (3 x) with

ether (30 ml). The remaining oil is dissolved in trifluoroacetic acid/anisole (4:1 v/v, 2 ml) and heated on a water bath, in a closed flask (25 ml) for 3 hours at 38°C.

The reaction mixture is then poured into dry ether, the organic solvents are decanted from the resulting slurry and

the remaining solids are washed with further portions of ether (3 x 30 ml). The crude, unprotected material is taken up in acetone/water (pH 2.5), extracted with ethyl acetate, the pH of the

the aqueous phase is adjusted to 7.6 with aqueous sodium hydroxide (2N) and extracted again with ethyl acetate. Concentration of the aqueous phase in vacuo to dryness gives a solid. This compound is characterized by acylation of the above sodium salt with 2',2',2'-trichloroethoxycarbonyl chloride using the method in

example IA, and one obtains 7-(2',2',2'-trichloroethoxycarboxamido)-3-(2-methyl-1,3,4-thiadiazol-5-ylthiomethyl)-4-(tetrazol-5-yl) -A -cephem

NMR (CDCl3)i 6 « 4.7(d) 1H, 4.2(m) 1H, 5.1(s) 2H, 5.35(s) 3H, 6.0(m) 2H and 7.2 (s) 3H.

Example 30

Sodium bicarbonate (42 g, 0.5 mol) is gradually added

to a vigorously stirred suspension of 7-amino-3-acetoxymethyl-A3--eefem-4-carboxylic acid (50 g, 0.18 mol) in acetone/water (250/500 mL).

A solution of benzyl chloroformate (36 g, 0.21 mol) in acetone (70 mL) is added dropwise to a stirred solution over 45 minutes. After stirring for 6 hours, acetone is removed using a rotary evaporator and the aqueous residue is washed with ethyl acetate to remove impurities. The aqueous solution is covered with ethyl acetate and acidified to pH 4.0. The combined ethyl acetate solutions from the extractions of the aqueous solution are washed with water and dried over magnesium sulfate. Evaporation of ethyl acetate gives the product 7-benzyloxycarboxamido-3-acetoxymethyl-A 3-cephem-4-carboxylic acid as an almost white solid (56 g), which is purified by thin-layer chromatography.

NMR (DMSOdg): δ 2.0(s) 3H, 3.5(s) 2H, 4.8-5.2(m) 5H and

7.3(s) 5H.

In a similar way, 7-benzyloxycarboxamido-3-(1-methyltrazol-5-ylthiomethyl)-A 3-cephem-4-carboxylic acid is prepared,

NMR (DMSOdg)s 6 « 7.5(s) 5B, 6.6(d) 1H, 6.2(s) 3B, 4.4(b)

2H, 4.04(s) 3H and 3.8(fl) 2R.

Example 31 7-benzyloxycarboxamido-3-acetoxymethyl-4-[N-(p-methoxybenzyl)-ftarbamoyl.- A 3-cephem

A solution of 2,4-dinitrophenol (22 g, 0.1 mol) in methylene chloride (100 ml) is added to a solution of 7-benzyloxycarbox-araido-3-acetoxymethyl-• A -3-cephem-4-carboxylic acid (41 g, 0.1mol) in dry dioxane (400 ml). To the stirred solution is added a solution of dicyclohexylcarbodiimide (21 g, 0.1 mol) in dioxane (100 ml). After 1.5 hours, the dicyclohexylurea is removed by filtration. To the filtrate is added a solution of p-methoxybenzyl-araine (13.7 g, 0.1 mol) in dioxane (ICO ml). After stirring for 6 hours, the precipitated solid material is filtered, washed with ether and ethyl acetate and dried, and the product is obtained as a white solid (30 g), which is purified by thin-layer chromatography.

NMR (DMSOdg): 6 - 2.0(s) 3H, 3.4(s) 2H, 3.7 (s) 3H, 4.3(d) 2H, 4.9(d) 2H, 5.0 (m) 4H, 5.4(q) 1H and 7.0(m) 9H*

7-Benzyloxycarboxamido-3-(1-methyl-tetrazol-5-ylthiometh<y>l)-4-[N-(p-methoxy<y>benz<y>l)carbamo<y>l] is prepared in a similar manner -A3 cofem.

NMR (CDCl 3 ); 6 » 7.4 (s) 5H, 7.1 (q) 4H, 7.7 (ia) 2H, 5.25 (s) 2H, 5.45 (d) lH, 4.5(m) 4H, 4.0(s) 3H, 5.9(s) 3H and 5.79(3) 2H.

Example 32

Phosphorus pentachloride (4.2 g, 20 mmol) is added to a suspension of 7-benzyloxycarboxamido-3-acetoxymethyl-4-[N-(p-methoxybenzyl)carbamoyl3-A 3-cephem (5.3 g, 10 mmol) in dry ethanol-free chloroform (100 mL) containing pyridine (1.7 g, 20 mmol). After 30 minutes, the nuclear magnetic resonance spectrum indicates that the reaction has ended.

The solution is cooled in an ice bath and a solution of tetramethylguanidinium azide (16.0 g, 100 mmol) in dry ethanol-free chloroform is added dropwise over 20 minutes. After a further 10 minutes, the ice bath is removed and stirring is continued for 30 minutes at ambient temperature. The solution is washed with water, sodium bicarbonate solution (3 x), 6N hydrochloric acid (3 x), water and dried over magnesium sulfate. Evaporation of the solvent gives the crude product as a brown solid (3.7 g), which is purified by column chromatography on silica. Elution with dichloromethanehexane (1:1) and then with ether:dichloromethanehexane (2:1) removes impurities. Elution with ether gives the product, 7-benzyloxy-carboxamido-3-acetoxymethyl-4-[1-(p-methoxybenzyl)tetrasol-5-yl3-α-cephem, as a white solid, which is purified by thin-screen chromatography.

NMR (DMSOdg): 6 « 1.9(s) 3H, 3.65(s) 2H, 3.8(s) 3H, 4.3(s) 2H, 5.15(s) 2H, 5.35 (d) 1H, 5.6(m) 3H, 7.2(m) 9H and 8.6(d) 1H.

In a similar way, 7-benzyloxycarboxamido-3-(1-methyltetrazol-5-ylthiomethyl)-4-[1-(p-methoxybenzyl)tetrazol-5-yl3-3 is prepared

A -cephem.

NMR (DMSOdg) in 6 » 7.3(s) 5H, 7.05(q) 4H, 6.5(s) 3H, 6.2(d) 1H, 5.05(s) 2H, 3.S5 (s) 3H, 3.7(s) 3H, 3.5-4 (m) 2H and 3.3(s) 2H„

Example 33

7- amino- 3- acetoxymethyl- 4-( tetrazol- S- yl)- A 3- cephem

7-Benzyloxycarboxamido-3-acetoxymethyl-4-[1-(p-methoxy-benzyl)tetrazol-5-yl3-A 3-cephem (100 mg) is dissolved in trifluoroacetic acid/anisole (4 ul v/v), (0.5 ml) and to this reddish-brown solution trifluoromethanesulfonic acid is added from a Pasteur pipette (25 drops) • Effervescence is immediately obtained and the solution becomes cherry red.

After 3 minutes at room temperature, the reaction mixture is stopped by the addition of ether (which has been dried down to sodium). A brown gum falls out, the overlying liquid is decanted and the residue is washed again with ether.

The viscous rubber dissolves again in the smallest possible volume

of methylene chloride containing 1% triethylamine and the solution is chromatographed on silica (2 mm, 20 x 20 Kieselgel 6 F2S4) using acetonitrile/water (6 ul v/v) as eluent.

The band at r.f. 0.4 was removed and extracted with 1% triethylamine/methylene chloride (3 x 150 mL). The organic extracts were evaporated to dryness in vacuo to give a white solid, a mixture of 7-amino-3-acetoxymethyl-4-(tetrasol-5-yl)-A<3->cephem and triethylamine hydrogen chloride.

NMR (CDCl 3 )s 6 o 5.25(d) 1H, 4.9 (ra) 2H and 2.05(s) 3H.

Example 34

7- amino- 3-( 1- methyltetrazol- 5- ylthiomethyl)- 4-( tetrazol- 5- yl)- A3~ cephem

Starting from 7-benzyloxycarboxamido-3-(1-methyltetrazol-5-ylthiomethyl)-4-[1-(p-methoxybenzyl)tetrazol»5-yl)-A 3=cephem as in example 33, one obtains the uncleaned product.

The zwitterion is isolated by crystallization of the impure reaction mixture, after sweeping with dry ether, from the smallest possible volume of potassium hydrogen phosphate pH 7 buffer. After the dissolution of the impure reaction mixture, the zwitterion precipitates as a creamy yellow solid and is purified by thin-layer chromatography.

NMR (DMSOdg): 6 » 5.1(d) 1H, 4.8(d) 1H, 4.3(q) 2H, 3„9(s) 3H and 3.7 (s) 2H.

Example 35

7-(2•,2*,20-trichloroethoxycarboxamido)-3-methyl-A2~cef©m-4- carboxylic acid

7-(2*,211,2 • -trichloroethoxycarboxamido)-3-methyl-A3-cephem-4-carboxylic acid (66.6 g) and N-hydroxysuccinimide (21.6 g) in dioxane (200 ml) at room temperature are treated with dicyclohexylcarbodiimide (35.6 g) and the mixture is stirred for 1 hour at room temperature. The mixture is filtered and the filtrate is evaporated under reduced pressure, and a solid oil is obtained which is dissolved in pyridine (400 ml) at 7°C and which is then treated with aqueous sodium hydroxide (7.5 g in 100 ml) with vigorous stirring. The mixture is allowed to warm to 25°C and stirring is continued for an additional 1.75 hours. The pyridine is evaporated under reduced pressure

and the rest is poured over ice. The pH is adjusted to 2 with 6N hydrochloric acid and

. the solution is extracted into ethyl acetate.' The organic solution is washed with IN hydrochloric acid, water and evaporated under reduced pressure. The residue is dissolved in aqueous sodium carbonate solution and the resulting solution is washed with ethyl acetate. The aqueous solution is treated. with charcoal, is filtered and the pH of the filtrate is adjusted to 2 med

6N hydrochloric acid. The skimming is extracted with ethyl acetate and the organic solution is treated with charcoal, filtered and dried (MgSO 4 ). The solution is then evaporated under reduced pressure, and

obtains the product as a foam, which solidifies on trituration with petroleum ether (48.0 g) and which is pure enough after chromatography.

NMR (CDCl 3 ): 6 - 7.5(d) 1H, 6.0(q) 1H, 5.2(m) 2H, 4.7(s) 2H, 4.6 (s) 1H and 1.9 (8) 3K.

Example 36 7-(2',2',2'trichloroethoxycarboxamido)-3-methyl-4-[N-(p-methoxybenzyl)carbamoyl3-A-cephem

7-(2*,2',2'-trichloroethoxycarboxamido)-3-methyl-A 2-cephem-4-carboxylic acid (46.0 g) and N-hydroxysuccinimide (14.3 g) in dioxane (750 ml) are treated after 1 hour with dicyclohexylcarbodiimide (24.3 g) at room temperature with stirring.To the stirred solution is added p-methoxybenzylamine (32.8 g) in dioxane (200 ral) and the mixture is stirred for a further 1 hour at room temperature.

The mixture is filtered and the solution is stirred for a further 1 hour at room temperature. The mixture is filtered and the solution is concentrated to 300 ml under reduced pressure. The concentrate is poured onto a saturated, aqueous NågCOj solution (2 litres) and the skimming is collected. The removal is washed with water, IN HC1 and

finally with water. The solid is dried under reduced pressure at 100°C for 116 hours, and 7-(2',2',2'-trichloroethoxycarboxamido)-3-methyl-4-[N-(p-methoxybenzyl)-carbamoyl3- A 2 -cephem (58.6 g). A sample recrystallized from acetone (m.p. 219-220°C). NMR (DMSOdg): 6 » 7.0(q) 4H, 6.0(q) 1H, 5.2(m) 2H, 4.7(s) 2H, 4.6(s) 1H, 4.2 (d) 2H, 3,7(s) 3H and 1,7(s) 3H.

Example 37 7-(2',2',2'-trichloroethoxycarboxamido)-3-methyl-4-[1r(p-methoxybenzyl)-tetrazol-5-yl3-A 2-cephem', i

7-(2',2',2•-trichloroethoxycarboxamido)-3-methyl-4-[N-(p-methoxybenzyl)carbamoyl3-A 2-pefem (44.2 g) and pyridine (14.06 g)

in chloroform (300 ml, ethanol-free) at 40°C is treated with phosphorus-

pentachloride (27.5 g) and the mixture is stirred at 40°C for 3 hours. The solution is cooled to room temperature and tetramethylguanidinium azide (87.9 g) is added, and the solution is stirred at room temperature for 2 hours. The solution is then washed with water, aqueous sodium bicarbonate, 1N hydrochloric acid, water and dried (MgSO 4 ). The organic solution is evaporated under reduced pressure, and a gum is obtained which is dissolved in ethyl acetate (200 ml) and which is passed through a column of aluminum oxide. The fraction containing the product is collected and the solution is evaporated to dryness under reduced pressure, and the product is obtained as a slightly orange colored solid (42.4 g).

NMR (CDCl 3 ): δ = 7.0(q) 4H, 6.1(d) 1H, 6.0(s) 1H, 5.6(q) 2H, 5.4(s) 1H, 5.2 (q) 1H, 4.8(d) 1H, 4.7(s) 2H, 3.7(s) 3H and 1.5(s) 3H.

Example 38

7-(2<«>,2<»>,2'-trichloroethoxycarboxamido)-3-methyl-4-(tetrasol-5-yl)-A2-e9Zem

7-(2',2•,2*-trichloroethoxycarboxamido)-3-methyl-4-11-(p-methoxybenzyl)tetrazol-5-yl]-A<2->cephem (40.7 g) in trifluoroacetic acid

(280 ml) containing anisole (70 ml) is allowed to stand at 50°C

for 6 hours. The solution is poured onto water (1 litre) and the suspension is extracted with ethyl acetate. The organic solution is extracted with saturated aqueous sodium bicarbonate and the aqueous solution is adjusted to pH 2 with 1N hydrochloric acid, and the supernatant is extracted with ethyl acetate. The organic solution is dried (MgSO4) and evaporated under reduced pressure, and 7-(2<*>,2',2<1->trichloroethoxycarboxamido)-3-methyl-4-(tetrazol-5-yl)- A 2-cephem as a faint brown solid (23.2 g). A sample recrystallized from chloroform (ethanol-free) m.p. 165-167°C.

NMR (DMSOdg): δ = 6.4(q* 1H, 5.8(s) 1H, 5.2(m) 2H, 4.8(s) 2H and 1.8(s) 3H).

Example 39 7-(2*,2*,2'-trichloroethoxycarboxamido)-3-methyl-4-(methoxymethyl-

2

tetrazol-5-yl)- A - cephem

To a suspension of 7-(2',2',2•-trichloroethoxycarboxamido)-3-methyl-4-(tetrazol-5-yl)-A<2->cephem (8.27 g) in methylene chloride (50 ml ) triethylamine (2.22 g) and chloromethyl methyl ether (1.76 g) are added and

the solution is stirred at room temperature for 30 minutes. The solution is then washed with water, aqueous sodium bicarbonate and dried (MgSOj. The evaporation under reduced pressure gives 7-(2',2',2'-trichloroethoxycarboxamido)-3-raethyl-4-(methoxymethyltetrazol-5-yl)-A 2-

cefem (7.3 g) as a cream-colored solid.

NMR-(CDCl1) s 6° 6.4(d) 1H, 6.0(q) 1H, 5.8(s) 2H, 4.7(s) 2H, 3.4(m) 3H and 1, 7(s) 3H.

Example 40

7- (211,2 • ,2 "-trichloroethoxycarboxamido)-3- (1-methyltetrazol-5=yl-thiomethyl)- 4-( methoxymethyltetrazol-5~ yl)- A - cephem

7-(2<*>,2",2'-trichloroethoxycarboxamido)-3-methyl-4-(methoxy-methyltetraaol-5-yl)-A 2-cephem (8.0 g) in carbon tetrachloride (160 ml) and chloroform (16 mL) was deoxygenated. N-bromosuccinimide (3.20 g) and benzoyl peroxide (200 mg) were added and the stirred mixture at 10-15°C was illuminated (250 watts, tungsten lamp) for 2 hours. During of this time additional chloroform was added to maintain a clear supernatant, and an additional amount of benzoyl peroxide (200 mg) was added 30 minutes after starting the illumination.

The succinimide was filtered off and the filtrate was treated with the triethylamine salt of 5-mercapto-1-methyltetrazole (3.40 g) for 2.5 hours at 45°C. The organic solution was then washed with water, aqueous sodium bicarbonate, water and finally dried (MgSOA). The organic solution was evaporated to dryness under reduced pressure. The residue was purified by means of column chromatography (silica gel, ethyl acetate, light petroleum ether, 1:1), and the product (3.0 g) is obtained.

NMR (CDCl 3 ): 6 « 6.6(s) 1H, 6.2(d) 1H, 6.0(s) 1H, 5.9(s) 2H, 5.4(m) 2H, 4.8 (s) 2H, 4.2(q) 2H, 4.0(s) 3H and 3.5(s) 3H.

Example 41

7-(2*,2*,2'-trichloroethoxycarboxamido)-3-(2-methyl-1,3,4-thiadiazol-5-ylthiomethyl)-4- methoxymethyltetrazol-5-yl)- A 2- cephem

7-(2',2',2'-trichloroethoxycarboxamido)-3-methyl-4-(methoxymethyltetrazol-5-ylthiomethyl)-A 2 -cephem (4.0 g) in carbon tetrachloride (60 ml) and chloroform (8 ml ) was treated with N-bromosuccinimide and benzoyl peroxide as in example 40. The product was treated in a similar way with the triethylamine salt of 5-mercapto-2-methylthiadiazole (2 g) g and one obtains, according to column chromatography! (silicon dioxide, ethyl acetate/light petroleum ether 1:1), 7-(2',2•,2*-trichloroethoxycarboxamido)-3-(2-methyl-1,3,4-thiadiazol-5-ylthiomethyl)-4-( methoxymethyltetrazol-5-yl)-A<2->cephem (1.1 g).

NMR (CDCl 3 ): 6 ■ 6.6 (a) 1H, 6.3(d) 1H, 6.1(s) 1H, 5.9 (s) 2H, 5.3-5.7 (m) 2H , 4.8(s) 2H, 4.1(q) 2H, 3.5(s) 3H and 2.7(s) 3H.

Example 42

7-(2',2',2•-trichloroethoxycarboxamido)-3-(1-arethyltetrazol~5-ylthiomethyl)-4-(methoxymethyltetrazol~5-yl)-A<2->cephem (2.0 g) in a'

mixture of acetic acid-water (7:3, 20 ml) at 0°C was treated with activated zinc dust (2.0 g) with stirring. After 1 hour, the mixture was filtered and the filtrate diluted with water (100 ml).

The pH of the mixture was adjusted to 2 with 1N hydrochloric acid and was then washed twice with ethyl acetate. The pH of the clear aqueous solution was adjusted to 6 with sodium bicarbonate and the mixture was extracted with chloroform. The organic solution was washed against pH 7 buffer, dried (MgSO^) and finally evaporated under reduced pressure, and 7~amino~3-(1-methyltetraaol-5-ylthiomethyl)-4-(methoxymethyltetrazol-5-yl) is obtained )-A 2-cephem (0.9 g) as a foam, which is purified by chromatography.

NMR (CDCl 3 ): δ 6.6(s) 1H, 5.95(s) 1H, 5.85(s) 2H, 5.25(d) 1H,

4.70(d) 1H, 4.1(q) 2H, 3.95(s) 3H and 3.5(s) 3H.

In a similar manner, 7-amino-3-(2-methyl-1,3,4-2-thiadiazol-5-ylthiomethyl)-4-(methoxymethyltetrazol-5-yl)-A-cephem is prepared.

NMR (CDCl3)s 6 » 6.8(s) 1H, €>.4(s) 1H, 6.0(s) 2H, 5.4(d) 1H, 4.8(d) 1H, 4, 2(q) 2H, 3.7(s) 3H and 3.2(s) 2H.

Example 43 7-(tetrazol-1-ylacetamido)-3-(2-methyl-1,3,4-thiadiazol-5-ylthiomethyl)-4-(tetrazol-5-yl)-A 3-cephem

A. 7-(tetrazol-1-ylacetamido)-3-(2-methyl-1,3,4-thiadiazol-5-yl-thlomethyl)-4-(methoxymethyltetrazol-5-yl)-A<2>-cephem

7-aroino-3-(2-methyl-1,3,4-thiadiazol-5-ylthiomethyl)-4-(methoxymethyltetrazol-5-yl)-A 2-cephem (430 mg) and tetrazol-1-yl-acetic acid (133 mg) in tetrahydrofuran (10 ml) and acetonitrile (10 ml) is treated with dlcyclohexylcarbodylimide (215 mg) and the mixture is allowed to stand at room temperature for 1 hour. The mixture is filtered and evaporated under reduced pressure, and 7-(tetrazol-1-ylacetamido)-3-(2-ethyl-1,3,4-thiadiaol-5-ylthiomethyl)-4-(methoxy-methyltetrazol-5- yl)~A 2-cephem (520 mg), as a cream-colored solid, which is purified by thin-layer chromatography!.

NMR (CDCl 3 ): 6 « 9.1(s) 1H, 6.6(s) 1H, 6.0(s) 1H, 5.9(s) 2H, 5.4(s) 2H, 5.4 (s) 1H, 5.2(d) 1H, 4.0(q) 2H, 3.4(s) 3H and 2.6(s) 3H.

B. 7-(tetrazol-1-ylacetamido)-3-(2-methyl-1,3,4-thiadiazol-5-ylthio-3^methyl)-4-(methoxymethyltetrazol-5-yl)rA^cefem-1 - oxide 7-(tetrazol-1-ylacetamido)-3-(2-methyl-1,3,4-thiadiazol-5-ylthiomethyl)-4-(methoxymethyltetrazol-5-yl)-A-recefem (500 mg) in chloroform (20 ml) is treated with m-chloroperbenzoic acid (219 mg) at 0°G with stirring. After 1 hour at 0°C, pyridine is added

(40 mg) and the mixture is allowed to warm to room temperature » After a further 1 hour, the mixture is concentrated under reduced pressure and diethyl ether is added to the concentrate. The cream-colored solid that precipitates is collected, dried under reduced pressure and the product (410 mg) is obtained as an ethyl-cream-colored solid, which is purified by chromatography.

NMR (DMSOdg/acetone dg) t 6°8.9(s) 1H, 6.0(s) 2H, 6.0, 1H, 5.6(s) 2H, 5.2(d) 1H, 4, 5(q) 2H, 4.0(s) 2H, 3.5(s) 3H and 2.7(s) 3H.

C. 7-(tetrazol-1-ylacetamido)-3-(2-methyl-1,3,4-thiadiazol-5-ylthio-methyl)- 4-(methocaylmethyltetrazol-5-yl)- A 3- cephem

7-(tetrazol-1-ylacetamido)-3-(2-methyl-1,3,4-thiadiazol?-5-ylthiomethyl)-4-(methoxymethyltetrazol-5-yl)-A 3-cephem-1-oxide ( 400 mg) in dimethylformamide (10 ml) is treated with anhydrous stannous chloride (400 mg) and acetyl chloride (200 mg) at 0°C with stirring. The mixture is stirred for 1 hour and then poured into an excess of water. The aqueous mixture is extracted several times with chloroform and the organic solution is washed several times with water.

The chloroform solution is dried (MgSO^) and evaporated under reduced pressure, and the compound is obtained as a foam (250 mg), which is purified by thin-layer chromatography1.

NMR (CDCl 3 ) 36 « 9.0(s) 1H, 8.7(d) 1H, 6.0(s) 2H, 6.0, 1H, 5.4(s) 2H, 5.2(d) 1H, 4,4(q) 2H, 3,7(s) 2H, 3,4(s) 3H and 2,7(s) 2H.

D. 7-(tetrazol-1-ylacetaraido)-3-(2-methyl-1,3,4-thiadiazol-5-ylthio-methyl)-4-(tetrazol-5-yl)- A 3- cephem

7-(tetrazol-1-ylacetamido)-3-(2-methyl-1,3,4-thiadiazol-5-ylthiomethyl)-4-(methoxymethyltetrazol-5-yl)-A-cephem (70 mg) in trifluoroacetic acid ( 2 ml) and anisole (0.5 ml) is allowed to stand at 20°C for 6 hours. After this, the trifluoroacetic acid is removed under reduced pressure and an oil is obtained. This residue is diluted with ethyl acetate and the resulting organic solution is extracted with

saturated aqueous sodium bicarbonate solution. The aqueous solution is acidified to pH 2 with 2N hydrochloric acid and the solution is extracted with ethyl acetate. The organic solution is dried (MgSO^) and evaporated under reduced pressure, and an oil is obtained which is crystallized by the addition of diethyl ether, and the product is obtained as colorless

needles (35 mg), which are purified by chromatography•

NMR (trifluoroacetic acid-d) t 6 «= 9.5 (o) 1H, 5.8 (d) 1H, 5.5 (s) 2H, 5.2(d) 1H, 4.5(s) 2H , 3.6(s) 2H and 2.8(s) 3H.

Example 44

7-D™ (α-hydroxy-α-phenylacetamido)-3-(1-methyltetrazol-5-ylthiomethyl)-4-(tetrazol-5-yl)- A 3- cephem

A. 7-D-(e-formyloxy-a-phenylacetamido)-3-(1-methyltetraaol-5-yl-thiomethyl)- 4-(roethoxymethyltetrazol-5-yl)- A 2- cephem

By using suitable reagents and applying the procedure in example 43A, the desired intermediate product is prepared"

NMR (CDCl3)t 6 «= 8.2 (s) 1H, 7.3(s) 5E, 6.4(s) 1H, 6.2(s) 1H, 5.9 (s) 1H, 5, 7(s) 2H, 5.5(q) 1H, 5.2(d) 1H, 4.0(q) 2H, 3.9(s) 3H, and 3.4(s) 3H.

B. 7-D-(ci-formyloxy-a-phenylacetamido)-3-(1-methyltetrazol-5-yl-thiomethyl)-4-(methoxymethyltetra20l-5-yl)- A 3-cephem-1^-oxide

By using the above intermediate from Example 44A and following the procedure in Example 43B, the desired compound is isolated and used in the following reaction for further purification.

C. 7-D-(ct-formyloxy-os-phenylacetamido)-3-(1-methyltetrazol-5-yl-thiomethyl)- 4-(methoxymethyltetrazol-5-yl)-A 3- cephem

The method in example 4 3C is used because the suitable compound from example 44B was used as the starting point, and the desired product is obtained.

NMR (CDCl3)s 6 = 8.3(s) 1H, 7.4(o) 5H, 6.3(s) 1H, 6.0(s) 2H, 6.0, 1H, 5.2(d ) 1H, 4.5(q) 2H, 4.0(s) 3H, 3.8(s) 2H and 3.5(s) 3H.

D. 7-D-(ci-formyloxy-ot-phenylacetamido)-3-(1-methyltetrazol-5-yl-

3

thiomethyl) - 4° (tetrazol-5-yl) - A - cephem

The trifluoroacetic acid-anisole reaction in example 43D is repeated starting from the product in example 44C, and the desired product is obtained. _

NMR (acetone-d6/D2O)t 6 » 8.3(s) 1H, 7.5(m) 5H, 6.2(s) 1H, 5.9(d) 1H, 5.3(d) 1H , 4.4(s) 2H and 4.0 (HOD).

(acetone-dg/D 2 O/trifluoroacetic acid-d)s 6 = 4.4(s) 2H, 4.0(s) 3H and 3.9 (s) 2H.

E. 7-D-(a-hydroxy-o-phenylacetamido)-3-(1-methyltetrazol-5-yltholo-3 methyl)-4-(tetrazol-5-yl)-A-cephem

7-D-(a-formyloxy-a-phenylacetamido)-3-(1-methyltetrazol-5-ylthiomethyl)-4-(tetrazol-5-yl)-A -cephem (100 mg) 1 aqueous sodium bicarbonate solution (5 ml) is allowed to stand at room temperature for 3 hours, the pH of the reaction mixture is adjusted to 2 with 2N hydrochloric acid and extracted with ethyl acetate. The organic solution

is dried (MgSO^) and evaporated under reduced pressure, and the desired product is obtained as a cream-coloured, solid material (35 rag)„ which, after purification by means of thin-layer chromatography, is mainly pure..

Example 45

7-D-(α-amino-ε-phenylacetamido)-3-(1-methyltetrazol-5-ylthiomethyl)-4-( tetrazol-5-yl)- A 3- cephem- trifluoroacetate

A. 7-[D-o-(t-butoxycarbonylamino)-phenylacetamido]-3-(1-methyl-tetrazol-5-ylthiomethyl)- 4-(methoxymethyltetrazol-5-yl)- A 2-cephem 7-amino-3- (1-methyltetrazol-5-ylthioraethyl)-4-(methoxymethyltetrazol-5-yl)-A<2>-cephem (100 mg) and N-t-butoxycarbonylphenylglycine (63.4 mg) in ethyl acetate-acetonitrile (1:1, 2 ml) is treated with dicyclohexylcarbodiimide (52 mg) at 15°C. After 1 hour, the mixture is filtered and the filtrate is diluted with ethyl acetate (10 ml) and the resulting solution is washed with 1N hydrochloric acid, aqueous sodium bicarbonate and finally with water. Drying (MgSO^) and evaporation under reduced pressure gives the product (152 mg) as a faint yellow solid.

NMR (CDCl 3 ): 6 « 7.25(s) 5H, 6.5(s) 1H, 6.0(s) 1H, 5.3(s) 2H, 5.6(m) 2H, 5.3 (m) 2H, 4.1(g) 2H, 3.9(s) 3H, 3.5(s) 3H and 1.4(s) 9H.

B. 7-[D-α™ (t-butoxycarbonylamino)phenylacetamido j ~ 3-(1-raethyl-

3 tetrazol-5-ylthiomethyl)-4-(methoxymethyltetrazol-5-yl)-A-cephem-1-oxide.

7-[D-ci-(t-butoxycarbonylamino)phenylacetamido]~3-(1-methyltetrazol-5-ylthiomethyl)-4-(methoxymethyltetrazol-5-yl)-A 2-cephem (140 mg, 0.22 mmol ) in chloroform (2 ml) is treated with m-chloroperbenzoic acid (51 mg, 85%) at 0°C with stirring. The solution is kept at 0°c for 1 hour and then pyridine (20 mg) is added and the solution is heated to room temperature and kept at that temperature for a further 2 hours.

The reaction mixture is diluted with chloroform and washed with 1N hydrochloric acid, aqueous sodium bicarbonate and dried (MgSO 4 ). Evaporation under reduced pressure gives the desired product as a cream solid (120 mg).

This material is used directly in the next reaction without further purification.

C. 7 " i D-a-(t-butoxycarbonylamino)phenylacetamido 3*-3-(1-methyltetrazol-5-yl-thiomatyl)-4-(methoxymethyltetrazol-S-yl)- A 3-cephem 7-[D-a- (t-butoxycarboxylarino)phenylacatamido3-3-(1-methyltetrazol-5-ylthiomethyl)-4-(methoxymethyltetrazol-5-yl)-α 3-cephem-1-oxide (120 mg) was suspended in dimethylformamide (0, 3 ml) and acetonitrile (0.75 ml) and the mixture is treated with acetyl chloride (60 mg) and anhydrous stannous chloride (33 mg) with stirring at 0°C

for 1 hour. The mixture is then allowed to warm to room temperature and stirred for a further 1 hour. The mixture is concentrated under reduced pressure and then diluted with ethyl acetate. The organic solution is washed with water and aqueous sodium bicarbonate and finally dried (MgSO 4 ). Evaporation under reduced pressure gives an oil which was purified by preparative thin-screen chromatography (silicon dioxide, ethyl acetate-light petroleum ether, 3:2), and the desired product is obtained as a faint yellow foam (60 mg).

NMR (CDCl 3 ): 6' = 7.4 (s) 5H, 7.2 (d) 1H, 5.8 (m) 4H, 5.3 (d) 1H, 5.2(d) 1H, 4, 5(q) 2H, 4.0(s) 3E, 3.8(s) 2H, 3.6(s) 3H and 1.5(s) 9H.

D. 7-D-(α-amino-α-phenylacetamido)-3-(1-methyltetrazol-5~ylthiomethyl-4-(tetrazol-5-yl)-α<3>~ceferg trifluoroacetate

7-tD-(a-t-butoxycarbonylamino-o-phenyl)acetamido]-3-(1-methyltetrazol-5-ylthiomethyl)-4-(methoxymethyltetrazol-5-yl)-A<3>-cephem (55 mg) in trifluoroacetic acid (2 ml) and aniseed (0.5 ml) are allowed to stand at 20°C for 6 hours. The trifluoroacetic acid is evaporated under reduced pressure and the residue is treated with ether. The resulting solid is collected and washed with portions of dry ether.

The solid is dried under high vacuum, and the desired product is obtained as a trifluoroacetic acid salt.

NMR (D2O/DMS0d6/trifluoroacetic acid©-d): 6 - 7.4(s) 5H, 5.8(d) 1H, 5.2(d) 1H, 5.1(s) 1H, 4.2( s) 2K, 4.0(3) 3H and 3.6(s) 2H.

Example 46

The following ingredients are mixed together in the specified amounts by weight.

After a good mixture has been achieved, it is prepared

tablets of the mixture and each tablet is of such a size as to contain 100 mg of the cephem compound.

Tablets containing respectively 50 and 250 mg of the active ingredient are also prepared, by selecting appropriate amounts of the cephem compound and the excipient mixture in each case.

Example 47

The following ingredients are mixed together in the indicated amounts by weight.

The well-mixed pharmaceutical mixture is filled into soft gelatin capsules, so that each capsule contains 100 mg of active ingredient.

Capsules containing respectively are also produced

50 and 250 mg of active ingredient by varying the amount of the cephem compound and the excipient mixture.

Example 48

The sodium salt of 7-(2-thienylacetamido)-3-methyl-4-(tetrazol-5-yl)-A-cephem is mixed well and ground with sodium citrate (4% by weight). The ground, dry mixture is sterilized and packaged in sterile,

small medicinal vials, which are sealed with serum capsules under sterile conditions* When this preparation is to be used, sufficient sterile water is injected into the small medicinal vials to dissolve the contents, and a solution containing 25 mg/ml of active ingredient is obtained. For parenteral use, the solution is withdrawn from the small medicinal vials using a syringe.

In a similar way, by varying the amount of added water, solutions containing 10, 50, 100 and 200 mg/ml of active ingredient are obtained respectively. Example 49 7-[D-(α-amino-α-phenyl)acetamido]-3-methyl-4-(tetrazol-5-yl)-α<3->cephem potassium salt

To a stirred solution of 1.94 g of 7-[D-(a-amino-a-phenyl)-acetamido]-3-methyl-4-(tetrazol-5-yl)-A 3-cephem in 100 ml of methanol, cooled to -30°C, a sufficient 1.0N solution of potassium hydroxide in methanol is added dropwise so that 1 equivalent of base is added. The mixture is allowed to warm to 0°C and is then added dropwise with stirring to 700 ml of ether. The solid that precipitates is removed by filtration and dried under high vacuum. This gives the potassium salt of the final product in good yield.

When the above procedure is repeated, except that the calcium hydroxide used is replaced by an equimolar amount of sodium hydroxide, the product is the sodium salt of 7-[D-(α-amino-α-phenyl)acetamido]-3-methyl-4-(tetrazole-5 -yl)-A 3-cephem.

Example 50

7-[D-a-amino-a-(p-hydroxyphenyl)acetamido]-3-methyl-4-(tetrazol-5-yl)- A<3>- cephem- calcium salt

To a stirred solution of 3.87 g of 7-[D-a-amino-a-(p-hydroxy-phenyl)acetamido]-3-methyl-4-(tetrazol-5-yl)-A 3-cephem in 40 ml of dimethylformamide a cloudy solution of 370 mg calcium hydroxide is added over the course of 5 minutes. The mixture is heated at 35-40°C for 1 hour and then a further 30 ml of dimethylformamide is added. The heating at 35-40°C is continued for a further 30 minutes, and the cooled solution is added dropwise to 700 ml of ether. An oil falls out. The solvent is decanted from and until the residue is added 100 ml of ethanol, followed by 400 ml of ether. The oil solidifies slowly and is then obtained by filtration and dried under high vacuum. This gives the calcium salt of the final product.

Example 51 7-[D-(α-amino-α-phenyl)acetamido]-3-methyl-4-(tetrazol-5-yl)-α<3->cephem hydrochloride salt

A suspension of 50 mg of 7-[D-(a-amino-a-phenyl)acetamido]-3-methyl-4-(tetrazol-5-yl)-A 3-cephem in 2 ml of deionized water is stirred for 5 minutes at ambient temperature. The pH is adjusted to 2.45 by using dilute hydrochloric acid and the resulting solution is immediately lyophilized. This gives the hydrogen chloride salt of the compound as a white solid.

Claims (1)

1. Method for producing a cephem derivative with formula and salts thereof; where T is a tetrazolyl precursor or a tetrazolyl group that has the formula wherein R is selected from the group consisting of (1) hydrogen (2) a nitrogen protecting group which is 2,2,2-trichloroethoxycarbonyl, 2,2,2-tribromomethoxycarbonyl, benzyloxycarbonyl, or wherein R 8 , R 1 and R 1 are each selected from the group consisting of hydrogen, chloro, bromo, fluoro, methyl, methoxy and phenyl and (3) an acyl group of the formula A is selected from the group consisting of hydrogen, acetoxy, 1-methyl-5-tetrazolylthio, 2-methyl-1,3,4-thiadiazolyl-5-thio and bromine, and 1*2 is a tetrazolyl cepheme nitrogen protecting group or forerunner thereof and is selected from the group consisting of where Rg is selected from the group consisting of alkyl containing from 1 to 3 carbon atoms and phenyl and R- is selected from the group consisting of hydroxy, methoxy, alkanoyloxy containing from 2 to 4 carbon atoms and benzyloxy and Rg is selected from the group that consists of hydrogen, hydroxy, fluorine, chlorine, bromine, iodine, methyl, methoxy, alkanoyloxy containing from 2 to 4 carbon atoms, phenyl and benzyloxy and wherein Rg and R^Q are each selected from the group consisting of hydrogen and methyl and X is selected from the group consisting of oxygen and sulfur; R^ is selected from the group consisting of hydrogen, alkanoyloxymethyl containing from 3 to 6 carbon atoms, 1-(alkanoyloxy)ethyl containing from 4 to 7 carbon atoms, methoxymethyl and phthalidyl, and R11 is selected from the group consisting of R1 and R2, Ar is selected from the group consisting of cyano, bromo, phenyl, mono- or disubstituted phenyl wherein each substituent is hydroxy, fluoro, chloro, bromo, amino, methoxy or methyl, phenoxy, phenylthio, pyridylthio, thienyl, 2-methyl-1,3,4-thiadiazol-5-ylthio, 1-tetrazolyl , alkyl containing from 1 to 12 carbon atoms, alkenyl containing from 2 to 12 carbon atoms, cycloalkyl containing from 3 to 7 carbon atoms, cycloalkenyl containing from 5 to 8 carbon atoms, cycloheptatrienyl, 1,4-cyclohexadienyl, 1-aminocycloalkyl containing from 4 to 7 carbon atoms, 5-methyl-3-phenyl-4-isoxazolyl, 5-methyl-3-(o-chlorophenyl)-4-isoxazolyl, 5-methyl-3-(2,6-dichlorophenyl)-4- isoxazolyl, 5-methyl-3-(2-chloro-6-fluorophenyl)-4-isoxazolyl, 2-alkyloxy-l-naphthyl having from 1 to 4 carbon atoms in alkoxy, sydnonyl, furyl, pyridyl, thiazolyl, isothiazolyl, pyridyl , thiazolyl, isothiazolyl, pyrimidinyl, triazolyl, imidazolyl, pyrazolyl, substituted phenoxy, substituted phenylthio, substituted pyridylthio, substituted thienyl, substituted furyl, substituted pyridyl, substituted tetraz olyl, substituted thiazolyl, substituted isothiazolyl, substituted pyrimidinyl, substituted triazolyl, substituted imidazolyl and substituted pyrazolyl wherein each substituted group is substituted with up to two groups selected from the group consisting of fluorine, chlorine, bromine, hydroxy, hydroxymethyl, amino, N ,N-dialkylamino where the alkyl groups contain from 1 to 4 carbon atoms, alkyl containing from 1 to 4 carbon atoms, aminomethyl, aminoethyl, alkoxy containing from 1 to 4 carbon atoms, alkylthio containing from 1 to 4 carbon atoms, 2-aminoethoxy and N- alkylamino containing from 1 to 4 carbon atoms, Q is selected from the group consisting of hydrogen, hydroxy, azido, amino and carboxy, n is an integer of 0 or 1, A is selected from the group consisting of hydrogen, acetoxy, 1-methyltetrazol-5-ylthio and 2-methyl-1,3,4-thiadiazol-5-ylthio, provided that when Ar is selected from the group consisting of pyridylthio , phenoxy, phenylthio, 2-methyl-1,3,4-thiadiazol-5-ylthio, bromo and cyano and n is equal to 1, Q is selected from the group consisting of hydrogen and carboxy, provided that R is and A is different from bromine and when A is bromine, T is a tetrazolyl group which, in the reaction sequence for its preparation, comprises at least one of the following steps: (1) acylation of a compound of the above formula, where, R is hydrogen and T is a tetrazolyl group, with one organic acylating agent and you get R equal to (2) transfers the amide group T which is to a tetrazolyl group such as T. (3) alkylates a compound where R is hydrogen or amino-protecting group, T is tetrazolyl and R^ or R^ is hydrogen, with a halide compound, to make R1 different from hydrogen? or (4) brominates one compound wherein R is hydrogen or an amino protecting group, T is tetrazolyl with R 1 or R^ -CH 2 -O-CH 3 , and A is hydrogen to produce a compound wherein A is bromine and, if desired, removes the methoxymethyl group . 2. Procedure as stated in claim 1, characterized in that a compound with the formula is produced where R is the acyl group T is the tetrazolyl group of the formula where R1 is hydrogen, alkanoyloxymethyl containing from 3 to 6 carbon atoms, 1-(alkanoyloxy)ethyl containing from 4 to 7 carbon atoms, methoxymethyl or phthalidyl, is R 1 or a tetrazolylcephem nitrogen protecting group, A is hydrogen, acetoxy, 1-methyl-5-tetrazolylthio or 2-methyl-1,3,4-thiadiazolyl-5-thio, characterized by acylating a compound of formula with an organic acid reacylating agent. 3. Procedure as stated in claim 2, characterized in that, after acylation of the amino group, any tetrazolyl cephem nitrogen protecting group is removed. 4. Procedure as stated in claim 2, characterized in that R^ or is hydrogen. 5. Procedure as stated in claim 2, characterized in that R 1 or R 1 is alkanoyloxymethyl, 1-(alkanoyloxy)ethyl, methoxymethyl or phthalidyl. 6. Procedure as stated in claim 2, characterized in that R^ is where R6 is hydrogen, alkyl containing from 1 to 3 carbon atoms or phenyl, E <_> is hydroxy, methoxy, alkanoyloxy containing from 2 to 4 carbon atoms or benzyloxy, and Rg is hydrogen, hydroxy, fluorine, chlorine, bromine, iodine, methyl, methoxy, alkanoyloxy containing from 2 to 4 carbon atoms, phenyl or benzyloxy, Rg and R^O are each hydrogen or methyl, and X is oxygen or sulfur. 7. Procedure as specified in claim 2. characterized in that A is hydrogen.. 8. Procedure as stated in claim 2, characterized in that A is acetoxy. 9. Procedure as stated in claim 2, characterized in that the organic acid acylating agent has the formula where Y^ is hydroxy, chlorine, bromine, a mixed anhydride group or an activated ester group and Ar, Q and n are as stated in claim 1. 10. Method as set forth in claim 1, characterized in that a compound of formula is prepared where. R is the amino protecting group as defined in claim 1, R is a tetrazolyl group of the formula where R 2 is the tetrazolyl cephem nitrogen-protecting group, A' is hydrogen, acetoxy, 1-methyl-5-tetrazolylthio or 2-methyl-1,3,4-thiadiazolyl-5-thio comprising the step for the transfer of the amide group of a compound with formula where R 2 is a potential tetrazolyl cepheme nitrogen protecting group, to the tetrazolyl group T. 11. Method as set forth in claim 10, characterized in that the tetrazolyl cepheme nitrogen protecting group is removed after the formation of the tetrazolyl ring. 12. Method as stated in claim 10, c characterized in that the amino protecting group is removed after the formation of the tetrazolyl ring. 13. Method as stated in claim 1, for production of a compound with the formula where R is hydrogen or the amino protecting group. A" is hydrogen, acetoxy, 1-methyl-5-tetrazolylthio or 2-methyl-1,3,4-thiadiazolyl-5-thio, T is a tetrazolyl group of the formula where R11 ' or R^^ ' is alkanoyloxymethyl containing from 3 to 6 carbon atoms, 1-(alkanoyloxy)ethyl containing from 4 to 7 carbon atoms, methoxymethyl or phthalidyl, characterized by alkylating the corresponding tetrazolyl compound where R,, 1 or R^ ' is hydrogen, with R^<*> halide. 14. Procedure as specified in claim 13, characterized in that an optional amino protecting group is removed after the alkylation. 15. Procedure as stated in claim 14, characterized in that R^' or R^' is methoxymethyl. 16. Method as stated in claim 1, for production of a compound of formula where R is hydrogen or the amino protecting group, T" is a tetrazolyl group with the formula characterized by brominating a compound with the formula and, if desired, then removes the methoxymethyl group. 17. Cephem derivative with formula and salts thereof; where T is a tetrazolyl precursor or a tetrazolyl group which is either wherein R is selected from the group consisting of (1) hydrogen (2) a nitrogen protecting group which is 2,2,2-trichloroethoxycarbonyl, 2,2,2-tribromomethoxycarbonyl, benzyloxycarbonyl, or wherein R^r and Rg are each selected from the group consisting of hydrogen, chlorine, bromine, fluorine, methyl, methoxy and phenyl and (3) a acyl group with the formula A is selected from the group consisting of hydrogen, acetoxy, 1-methyl-5-tetrazolylthio, 2-methyl-1,3,4-thiadiazolyl-5-thio and bromine, and R 2 is a tetrazolyl cepham nitrogen protecting group or forerunner thereof and is selected from the group consisting of where Rg is selected from the group consisting of alkyl containing from 1 to 3 carbon atoms and phenyl and R- is selected from the group consisting of hydroxy, methoxy, alkanoyloxy containing from 2 to 4 carbon atoms and benzyloxy and Rg is selected from the group that consists of hydrogen, hydroxy, fluorine, chlorine, bromine, iodine, methyl, methoxy, alkanoyloxy containing from 2 to 4 carbon atoms, phenyl and benzyloxy and wherein Rg and R1Q are selected from the group consisting of hydrogen and methyl and X is selected from the group consisting of oxygen and sulfur. is selected from the group consisting of hydrogen, alkanoyloxymethyl containing from 3 to 6 carbon atoms, 1-(alkanoyloxy)ethyl containing from 4 to 7 carbon atoms, methoxymethyl and phthalidyl, and R^^ is selected from the group consisting of R^ and R 2 i Ar is selected from the group consisting of cyano, bromo, phenyl, mono- or disubstituted phenyl wherein each substituent is hydroxy, fluoro, chloro, bromo, amino, methoxy or methyl, phenoxy, phenylthio, pyridylthio, thienyl, 2-methyl-1,3,4-thiadiazol-5-ylthio, 1-tetrazolyl , alkyl containing from 1 to 12 carbon atoms, alkenyl containing from 2 to 12 carbon atoms, cycloalkyl containing from 3 to 7 carbon atoms, cycloalkenyl containing from 5 to 8 carbon atoms, cycloheptatrienyl, 1,4-cyclohexadienyl, 1-aminocycloalkyl containing from 4 to 7 carbon atoms, 5-methyl-3-phenyl-4-isoxazolyl, 5-methyl-3-(o-chlorophenyl)-4-isoxazolyl, 5-methyl-3-(2,5-dichlorophenyl)-4-isoxazolyl . thiazolyl, isothiazolyl, pyrimidinyl, triazolyl, imidazolyl, pyrazolyl, substituted phenoxy, substituted phenylthio, substituted pyridylthio, substituted thienyl, substituted furyl, substituted pyridyl, substituted tetrazolyl, substituted thiazolyl, substituted isothiazolyl, substituted pyrimidinyl, substituted triazolyl, substituted imidazolyl, and substituted pyrazolyl wherein each substituted group is subs titrated with up to two groups selected from the group consisting of fluorine, chlorine, bromine, hydroxy, hydroxymethyl, amino, N,N-dialkylamino wherein the alkyl groups contain from 1 to 4 carbon atoms, alkyl containing from 1 to 4 carbon atoms, aminomethyl, aminoethyl, alkoxy containing from 1 to 4 carbon atoms, alkylthio containing from 1 to 4 carbon atoms, 2-aminoethoxy and N-alkylamino as in holds from 1 to 4 carbon atoms, Q is selected from the group consisting of hydrogen, hydroxy, azido, amino and carboxy, n is an integer equal to 0 or 1, A is selected from the group consisting of hydrogen, acetoxy, 1-methyltetrazol-5-ylthio and 2-methyl-1,3,4-thiadiazol-5-ylthio, provided that when Ar is selected from the group consisting of pyridylthio , phenoxy, phenylthio, 2-methyl-1,3,4-thiadiazol-5-ylthio, bromo and cyano and n is equal to 1 Q is selected from the group consisting of hydrogen and carboxyl under the assumption that R is T is equal and A different from bromine, and when A is bromine, T is a tetrazolyl group. 18. Connection as stated in claim 17, characterized in that it is selected from the group consisting of where R. is selected from the group consisting of 2,2,2-trichloroethoxycarbonyl, 2,2,2-tribromomethoxycarbonyl, benzyloxycarbonyl, hydrogen and wherein R 3 , R 4 and R 8 are selected from the group consisting of hydrogen, chlorine, bromine, fluorine, methyl, methoxy and phenyl; A is selected from the group consisting of hydrogen, acetoxy, 1-methyl-5-tetrazolylthio and 2-methyl-1,3,4-thiadiazolyl-5-thio, and R2 is selected from the group consisting of wherein Rg is selected from the group consisting of alkyl containing from 1 to 3 carbon atoms and phenyl and R„ is selected from the group consisting of hydroxy, methoxy, alkanoyloxy containing from 2 to 4 carbon atoms and benzyloxy and Rg is selected from the group that consists of hydrogen, hydroxy, fluorine, chlorine, bromine, iodine, methyl, methoxy, alkanoyloxy containing from 2 to 4 carbon atoms, phenyl and benzyloxy and wherein Rg and R^Q are each selected from the group consisting of hydrogen and methyl and X is selected from the group consisting of oxygen and sulfur. 19. Connection as stated in claim 18, characterized in that A is selected from the group consisting of hydrogen and acetoxy. 20. Compound as stated in claim 19, characterized in that R 2 is p-methoxybenzyl and A is hydrogen. 21. Commitment as stated in claim 20, characterized in that the compound is 7-(2, ,2',2'-trichloroethoxycarboxamido)-3-methyl-4-[N-(p-methoxybenzyl)carbamoyl]-A <3-> cephem. 22. Connection as stated in claim 20, characterized in that the compound is 7-(benzyloxy-carboxamido)-3-methyl-4-[N-(p-methoxybenzyl)carbamoyl]-A 3-cephem. 23. Connection as stated in claim 20, characterized in that the compound is 7-(N-triphenyl-methylamino)-3-methyl-4-[N-(p-methoxybenzyl)carbamoyl3-A 3 -cephem. 24. Connection as stated in claim 20, characterized in that A is acetoxy and R2 is p-methoxybenzyl. 25. Connection as stated in claim 24, characterized in that the compound is 7-(2 <*> ,2',2'-trichloroethoxycarboxamido)-3-acetoxymethyl-4-[N-(p-methoxybenzyl)-carbamoyl]-A <3-> cephem. 26. Connection as stated in claim 24, characterized in that the compound is 7-(benzyloxy-carboxamido)-3-acetoxy-methyl-4-[N-(p-methoxybenzyl)carbamoyl]-A 3-cephem. 27. Connection as stated in claim 24, characterized in that the compound is 7-(N-triphenyl-methylamino)-3-acetoxy-methyl-4-[N-(p-methoxybenzyl)carbamoyl]-A 3-cephem. 28. Connection as stated in claim 24, characterized in that the compound is 7-(2',2',2'-trichloroethoxycarboxyaido)-3-methyl-4-[N-(p-methoxybenzyl)-carbamoyl]-A <2-> cephem. 29. Compound as stated in claim 17, v characterized in that it is selected from the group consisting of and salts thereof where R is selected from the group consisting of 2,2,2-trichloroethoxycarbonyl, 2,2,2-tribromomethoxycarbonyl, benzyloxycarbonyl, hydrogen and where R^i R^ and R^ are selected from the group consisting of hydrogen, chlorine, bromine, fluorine, methyl, methoxy and phenyl A is selected from the group consisting of hydrogen, acetoxy, 1-methyl-5-tetrazolylthio and 2-methyl-1,3,4-thiadiazolyl-5-thio, R^ is selected from the group consisting of hydrogen, alkanoyloxymethyl containing from 3 to 6 carbon atoms, 1-(alkanoyloxy)-ethyl containing from 4 to 7 carbon atoms, methoxymethyl and phthalidyl, and R^ is selected from the group consisting of hydrogen, alkanoyloxymethyl containing from 3 to 6 carbon atoms, 1-(alkanoyloxy)ethyl containing from 4 to 7 carbon atoms, methoxymethyl, phthalidyl, wherein Rg is selected from the group consisting of alkyl containing from 1 to 3 carbon atoms and phenyl and R- is selected from the group that consists of hydroxy, methoxy, alkanoyloxy containing from 2 to 4 carbon atoms and benzyloxy and Rg is selected from the group consisting of hydrogen, hydroxy, fluorine, chlorine, bromine, iodine, methyl, methoxy, alkanoyloxy containing from 2 to 4 carbon atoms, phenyl and benzyloxy and wherein Rg and R1Q are each selected from the group consisting of hydrogen and methyl and X is selected from the group consisting of oxygen and sulfur. 30. Compound as stated in claim 29 with formula I, characterized in that A is hydrogen, acetoxy, 1-methyl-5-tetrazolylthio and 2-methyl-1,3,4-thiadiazolyl-5-thio and R11 is P"methoxyben3y:1- • 31. Connection as stated in claim 30, characterized in that the compound is 7-(2',2',2 trichloroethoxycarboxamido)-3-methyl-4-[1-(p-methoxybenzyl)-tetrazol-5-yl]-A <3-> cephem. 32. Connection as stated in claim 30, characterized in that the compound is 7-(N-triphenyl- methylamino)-3-methyl-4-[1-(p-methoxybenzyl)tetrazol-5-yl]-a 3-cephem. 33. Connection as stated in claim 30, characterized in that the compound is 7-amino-3-methyl-4-[1-(p-methylbenzyl)-tetrazol-5-yl]-a 3-cephem. 34. Connection as stated in claim 30, characterized in that the compound is 7-(benzyloxy-carboxamido)-3-acetoxy-methyl-4-[1-(p-methoxybenzyl)tetrazol-5-yl]-A <3-> cephem. 35. Connection as stated in claim 30, characterized in that the compound is 7-(2',20,2'-trichloroethoxycarboxamido)-3-acetoxymethyl-4-[1-(p-methoxybenzyl)-tetrazol-5-yl]-A -cephem. 36. Connection as stated in claim 30. characterized in that the compound is 7-amino-3-acetoxymethyl-4-[1-(p-methoxybenzyl)tetrazol-5-yl]-a 3-cephem. 37. Connection as stated in claim 30, characterized in that the compound is 7-(benzyloxy-carboxamido)-3-(1-methyltetrazol-5-ylthiomethyl)-4-[1-(p-methoxy-3 benzyl)tetrazol-5-yl]-A -cephem. 38. Connection as stated in claim 30, characterized in that the compound is 7-(2',2 <0>,2°-trichloroethoxycarboxamido)-3-(2-methyl-1,3,4-thiadiazol-5-ylthio-methyl)-4-[1-( p-methoxybenzyl)-tetrazol-5-yl]-A 3-cephem. 39. Connection as stated in claim 30, characterized in that the compound is 7-amino-3-(2-methyl-1,3,4-thiadiazol-5-ylthiomethyl)-4-[1-(p-methoxybenzyl)-tetrazol-5-yl]-A 3- cephem. 40. Connection as stated in claim 30, characterized in that the compound is 7-(2',21,2'-trichloroethoxycarboxamldo)-3-methyl-4-[1-. ( p -Methoxybenzyl)-tetrazol-5-yl]- A <2> -cephem. 41. Connection as stated in claim 30, characterized in that the compound is 7-(benzyloxy-carboxamido)-3-(2-ethyl-1,3,4-thiadiazol-5-ylthiomethyl)-4-[1-(p-methoxybenzyl)tetrazole-.5 -.yl]-A -cephem.. 42. Connection as stated in claim 29, characterized in that R. and R.sub.3 are each hydrogen. 43. Connection as stated in claim 42, characterized in that the compound is 7-amino-3-methyl-4-(tetrazol~ 5-yl)-A 3-cephem. 44. Connection as stated in claim 42,... characterized in that the compound is 7-amino-3-acetoxymethyl-4-(tetrazol-5-yl)-A-cephem. 45. Compound as stated in claim 42, characterized in that the compound is 7-amino-3- (2-methyl-1,3,4-thiadiazol-5-ylthiomethyl)-4-(tetrazol-5-yl)-A <3-> cephem. 46. Connection as specified in claim 42, - characterized in that the compound is 7-amino-3- (1-methyltetrazol-5-ylthiomethyl)-4-(tetrazol-5-yl) - A 3-cephem. 47. Connection as stated in claim 42, characterized in that the compound is 7-(2',2',2,'-trichloroethoxycarboxamido)-3-methyl-4-(tetrazol-5-yl)-A <2-> cephem. 48. Connection as stated in claim 29, characterized in that R^ and R^ are each methoxymethyl. 49. Compound as stated in claim 48, characterized in that the compound is 7-(2',2•,2,'-trichloroethoxycarboxamido)-3-methyl-4-[1-(methoxymethyl)tetrazol-5-yl]-A 2-cephem and 7-(2*,2',2,-,trichloroethoxycarboxamido)f■■3-methyl-4-[2-(methoxymethyl)tetrazol-5-yl]-A 2-cephem. 50. Connection as stated in claim 48, characterized in that the compound is 7-amino-3-methyl-4-[1-methoxymethyl)-tetrazol-5-yl]-A <2-> cephem. 51. Connection as stated in claim 48, characterized in that the compound is 7-amino-3-methyl-4-[2-(methoxymethyl)-tetrazol-5-yl]-A 2-cephem. 52. Connection as stated in claim 48, characterized in that the compound is 7-amino-3-acetoxymethyl-4-[1-(methoxymethyl)tetrazol-5-yl3-A-cephem, 53. Compound as stated in claim 48, characterized in that the compound is 7-amirio-3-. acetoxymethyl-4-[1-(methoxymethyl)tetrazol-5-yl]-A<2> -cephem. 54. Compound as stated in claim 48, characterized in that the compound is 7-amino-3-(1-methyltetrazol-5-ylthiomethyl)-4-[1-(methoxymethyl)tetrazol-5-yl]-A 2-cephem. 55. Connection as stated in claim 48, characterized in that the compound is 7-amino-3-1-methyltetrazol-5-ylthiomethyl)-4-[2-(methoxymethyl)tetrazol-5-yl]-A 2 - cephem. 56. Connection as stated in claim 48, character! characterized in that the compound is 7-amino-3-(2-methyl-1,3,4-thiadiazol-5-ylthiomethyl)-4-[1-(methoxymethyl)tetrazol-5-yl]-A <2> -c ■efem <.> 57o Compound as stated in claim 48, characterized in that the compound is 7-amino-3-(2- methyl-1,3,4-thiadiazol-5-ylthiomethyl)-4-[2-(methoxymethyl)tetrazol-5-yl]-A <2-> cephem. 58. Compound as stated in claim 48, characterized in that the compound is 7-(2',2',2'-tribromomethoxycarboxamido)-3-methyl-4-[1-(methoxymethyl)tetrazol-5-yl]-A < 2-> cephem. 59. Compound as stated in claim 48, characterized in that the compound is 7-(2',2*,2'-tribromomethoxycarboxamido)-3-methyl-4-[2-(methoxymethyl)tetrazol-5-yl]-• A 2 -cephem.... 60. Connection as stated in claim 17, characterized in that the connection is selected from the group consisting of wherein R is selected from the group consisting of 2,2,2-tri(borethoxycarbonyl, 2,2,2-tribromomethoxycarbonyl, benzyloxycarbonyl, hydrogen and where R^ t R^ and R^ are selected from the group consisting of hydrogen, chlorine, bromine, fluorine, methyl, methoxy and phenyl, R^ is selected from the group consisting of hydrogen, alkanoyloxymethyl containing from 3 to 6 carbon atoms, 1-(alkanoyloxy)ethyl containing from 4 to 7 carbon atoms, methoxymethyl and phthalidyl, and is selected from the group consisting of hydrogen, alkanoyloxymethyl containing from 3 to 6 carbon atoms, 1-(alkanoyloxy)ethyl containing from 4 to 7 carbon atoms, methoxymethyl, phthalidyl, where Rb is selected from the group consisting of alkyl containing from 1 to 3 carbon atoms and phenyl and R- is selected from the group consisting of hydroxy, methoxy, alkanoyloxy containing from 2 to 4 carbon atoms and benzyloxy and Rg is selected from the group consisting of hydrogen, hydroxy, fluorine, chlorine, bromine, iodo, methyl, methoxy, alkanoyloxy containing from 2 to 4 carbon atoms, phenyl and benzyloxy and wherein Rg and R^Q are each selected from the group consisting of hydrogen and methyl and X is selected from the group consisting of oxygen and sulfur 61c Compound as stated in claim 60, characterized in that R is 2,2,2-trichloroethoxycarbonyl. 62o Connection as stated in claim 60, characterized in that the compound is 7-(2",2',2'-trichloroethoxycarboxamido)-3-bromomethyl-4-[1-(methoxymethyl)-tetrazol-5-yl]-A <2-> cephem. 63. Connection as stated in claim 60, characterized in that the compound is 7-(2',20,2 trichloroethoxycarboxamido)-3-bromomethyl-4-[2-(methoxymethyl)-tetrazol-5-yl]-A <2-> cephem.