BE831787A - NEW CEPHALOSPORIN DERIVATIVES - Google Patents

Description

       

   <EMI ID = 1.1>

  
The present invention relates to a new class of cephalosporin compounds having in position 3 a group-

  
 <EMI ID = 2.1>

  
a process for their preparation. These compounds exhibit a broad spectrum of gram-positive and gram-negative antibiotic activity.

  
The present invention provides novel cephalosporin compounds of formula I

  

 <EMI ID = 3.1>


  
 <EMI ID = 4.1>

  
inferior ; R2 is hydrogen, an alkali metal cation, or an easily removable ester-forming group; and R is hydrogen or the group

  
0

  
not

  
R'-C- wherein R 'is: hydrogen; an alkyl group in

  
 <EMI ID = 5.1>

  

 <EMI ID = 6.1>


  
formula in which A is the diphenylmethyl group, p-

  
 <EMI ID = 7.1>

  
methoxybenzyl and A 'is a C2-C4 alkanoyl, C2-C4 haloalkanoyl, benzoyl, halobenzoyl, 2,4-dinitrophenyl, or phthaloyl group;

  
or R 'is a group of formula
 <EMI ID = 8.1>
  <EMI ID = 9.1> halogen, hydroxyl or aminomethyl;

  
Z is 0 or S; and

  
m is equal to 0 or 1:

  
or R 'is a group of formula

  

 <EMI ID = 10.1>


  
formula in which P is 2-thienyl, 3-thienyl,

  
 <EMI ID = 11.1>

  

 <EMI ID = 12.1>


  
formula wherein a and a 'are as defined above; and

  
Q is the hydroxyl, formyloxy, acetoxy, carboxy, sulfo, amino, or -NHY group in which Y is the benzyloxycarbonyl, t-butyloxycarbonyl group

  
 <EMI ID = 13.1>

  
 <EMI ID = 14.1> <EMI ID = 15.1> it

  
0

  
 <EMI ID = 16.1>

  
2-thienyl group; 3-thienyl; 2-furyl; 2-oxazyl; 2thiazyl; 1-tetrazyl; benzotriazolyl; 1,3,4-thiadiazolyl-2thio; 1,2,5-thiadiazolyl-3-thio; 1,3,4-oxadiazolyl-2-thia;

  
 <EMI ID = 17.1>

  
triazolyl.

  
The present invention also provides a process for the preparation of the cephalosporin compounds of formula I, which comprises:

  
 <EMI ID = 18.1>

  
formula II
 <EMI ID = 19.1>
 formula wherein R and R2 are as defined above, with a triazinylthio derivative of formula.

  

 <EMI ID = 20.1>


  
 <EMI ID = 21.1>

  
the acylation of the compounds thus obtained in which R is hydrogen or Q is the amino group;

  
and if desired, removing the protecting groups from the amino group and / or the carboxyl group.

  
 <EMI ID = 22.1>

  
laughing "means an alkyl group containing from 1 to 4 atoms

  
carbon, for example, the methyl, ethyl, n-propyl, isopropyl, n-butyl, secondary butyl or isobutyl group. Preferably, R- is a methyl or ethyl group and, more particularly

  
 <EMI ID = 23.1>

  
Examples of resulting substituents at position 3 of the

  
 <EMI ID = 24.1>

  
 <EMI ID = 25.1>

  
3-ylthiomethyl; 4-methyl-5-oxo-6-hydroxy-4,5-dihydro-1,2,4-. triazin-3-ylthiomethyl; 4-ethyl-5-oxo-6-hydroxy-4,5-dihydro-1,2, 4-triazin-3-ylthiomethyl; 4-n-propyl-5-oxo-6-hydroxy-4,5dihydro-1,2,4-triazin-3-ylthiomethyl; 4-isopropyl-5-oxo-6-

  
 <EMI ID = 26.1>

  
ylthiomethyl.

  
The R2 group of formula I is hydrogen, an alkali metal cation, such as lithium, sodium or potassium, <EMI ID = 27.1>

  
or'a group forming an easily removable ester. The term "easily removable ester-forming group" refers to the carboxylic acid protecting groups usually used and which are used to block the

  
 <EMI ID = 28.1>

  
sporin. Such groups are easily removed by conventional techniques, and they include, for example,

  
 <EMI ID = 29.1>

  
methoxybenzyl, 2,2,2-trichloroethyl, phenacyl, trimethylsilyl,

  
 <EMI ID = 30.1>

  
dissociable identical.

  
Examples of tert-C 1 -C 8 alkyl groups include, for example, t-butyl, t-amyl, and t-hexyl. Of

  
 <EMI ID = 31.1>

  
t-pentenyl, and t-hexenyl. Examples of tertio-

  
 <EMI ID = 32.1>

  
When R2 is a group giving an easily removable ester, it is preferred that it be the t-butyl group,

  
 <EMI ID = 33.1>

  
trichloroethyl, trimethylsilyl, acetoxymethyl, pivaloyloxymethyl, or phthalidyl. Preferably, when R2 is a group giving an easily removable ester, it is the '[pound] nitrobenzyl, acetoxymethyl or pivaloyloxymethyl group.

  
However, it is more particularly preferred that R2 is hydrogen or an alkali metal cation. These substituents of

  
 <EMI ID = 34.1>

  
on the antibiotic level.

  
Cleavage of the ester moiety to the free 4-carboxylic function is desirable to give a cephalosporin in which R2 is hydrogen or an alkali metal cation. The cut is made by conventional treatment. This includes, for example, treating the ester with an acid such as trifluoroacetic acid, or hydrochloric acid, or with zinc and an acid, such as formic acid, acetic acid. or hydrochloric acid. In the same way, the cleavage can be carried out by hydrogenation of the ester in the presence of palladium, rhodium or one of their compounds, in suspension or on a support such as barium sulfate, carbon or alumina.

  
In the previous definition of the group R 'the term <EMI ID = 35.1>

  
n-amyl, isoamyl, n-hexyl and 2,3-dimethylbutyl. The term

  
 <EMI ID = 36.1>

  
chloromethyl, bromomethyl, 2-iodoethyl, 2-chloropropyl and 3-bromopropyl group. The term "cyanoalkyl group in

  
 <EMI ID = 37.1>

  
methyl, 2-cyanoethyl, 3-cyanopropyl and 2-cyanopropyl. The term

  
 <EMI ID = 38.1>

  
such as the azidomethyl, 2-azidoethyl, 3-azidopropyl group,

  
 <EMI ID = 39.1>

  
refers to groups such as hydroxymethyl, 2-hydroxyethyl, 3-hydroxypropyl, and 2-hydroxypropyl.

  
In the definition of the residue of substituted 4-amino ester-

  
 <EMI ID = 40.1>

  
C2-C4 nuclei "," C2-C4 haloalkanoyl ", and" halobenzoyl ". The term" C2-C4 alkanoyl group "refers to the acetyl, propionyl, or butyryl group. The term" haloalkanoyl group "

  
 <EMI ID = 41.1>

  
2-chloropropionyl, or 3-bromobutyryl. The term "halobenzoyl group" refers to chloro- and bromosubstituted benzoyl groups such as 4-chlorobenzoyl, 4-bromobenzoyl, and 2,4-dichlorobenzoyl.

  
As used herein, the term "halogen" and the term "halo" each refer to fluoro, chloro, bromo and iodo radicals.

  
 <EMI ID = 42.1>

  
lower straight chain and branched chain alkyls such

  
that the methyl, ethyl, n-propyl, isopropyl, n-butyl group

  
 <EMI ID = 43.1>

  
methoxy, ethoxy, isopropoxy, or n-butoxy group.

  
The following groupings are illustrative groupings of the grouping

  
0

  
and

  
 <EMI ID = 44.1>

  
formamido, acetamido, pro-

  
pionamido, butyramido, a-methylpropionamido, valeramido,

  
 <EMI ID = 45.1> <EMI ID = 46.1>

  
chlorobenzamido) valeramido.

  
The following groupings are illustrative groupings of 0

  
not

  
R'-C-NH in the previous definition in which R 'is

  

 <EMI ID = 47.1>


  
and in which m is equal to zero: the grouping

  
 <EMI ID = 48.1>

  
When, in the preceding formula, m is equal to 1 and Z represents
-0-, illustrative groups include the following groupings <EMI ID = 49.1>

  
When, in the preceding formula, m = 1 and Z represents S, illustrative groups include the following groups:

  
 <EMI ID = 50.1>

  
 <EMI ID = 51.1> <EMI ID = 52.1>

  
When R 'represents a group of formula P-CH-, the

  
Q 0 illustrative groups exhibiting the overall formula R'-C-NH include: the mandelamido group of formula

  

 <EMI ID = 53.1>


  
its O-formyl and O-acetyl derivatives represented by the general formula

  

 <EMI ID = 54.1>


  
formula in which T is hydrogen or a methyl group;

  
 <EMI ID = 55.1>

  

 <EMI ID = 56.1>


  
 <EMI ID = 57.1>

  

 <EMI ID = 58.1>


  
 <EMI ID = 59.1>
 <EMI ID = 60.1>
 <EMI ID = 61.1>

  
by the formula

  

 <EMI ID = 62.1>


  
formula in which Y is the benzyloxycarbonyl, tbutyloxycarbonyl group,

  
 <EMI ID = 63.1>

NH

  
example, the phenyl, halophenyl, furyl, monomethylamino, dimethylamino, monoethylamino, diethylamino, methylethyl group

  
 <EMI ID = 64.1>

  
amino, or diphenylamino. In each of the preceding formulas, R "'can be hydrogen or a C1-C3' alkyl group plus

  
 <EMI ID = 65.1>

  
 <EMI ID = 66.1>

  
they are attached, forming a heterocyclic ring such that Y has the structure:

  

 <EMI ID = 67.1>


  
 <EMI ID = 68.1>

  
Also included are 2-thienylacetamido, 3thienylacetamido, and 1-tetrazylacetamido "-substituted groups in which, in the preceding formula, the phenyl group is replaced.

  
by a 2-thienyl, 3-thienyl or 1-tetrazyl ring.

  
Illustrative examples of the above acetamido groups are the <EMI ID = 69.1> group

  
 <EMI ID = 70.1>

  
acetamido.

  
 <EMI ID = 71.1> previous definition in which R 'is R "-CH2- are the following groups:

  
 <EMI ID = 72.1>

  
 <EMI ID = 73.1>

  
represented by the following formula III:

  

 <EMI ID = 74.1>


  
 <EMI ID = 75.1>

  
fications as defined above. Illustrative examples of these preferred compounds shown in the form of their free acid are as follows:

  
 <EMI ID = 76.1>

  
carboxylic

  
7- (2,5-dichlorophenylthioacetamido) -3- (4-isopropyl- acid

  
 <EMI ID = 77.1>

  
4-carboxylic, <EMI ID = 78.1>

  
 <EMI ID = 79.1>

  
is that represented by formula IV

  

 <EMI ID = 80.1>


  
formula in which R "represents the 2-thienyl group,

  
 <EMI ID = 81.1>

  
meaning as defined above. Illustrative examples of the foregoing compounds represented by formula IV and presented in their free acid form are as follows:

  
7- (2-thienylacetamido) -3- (4-methyl-5-oxo-6-hydroxy-4 acid,

  
 <EMI ID = 82.1>

  
 <EMI ID = 83.1>

  
the one represented by the following formula V
 <EMI ID = 84.1>
  <EMI ID = 85.1>

  
phenyl, or substituted phenyl as defined below and Q is a hydroxyl, formyloxy, acetoxy, carboxy or amino group. Illustrative examples of the foregoing compounds represented by formula V and presented in their free acid form are as follows:

  
 <EMI ID = 86.1>

  
 <EMI ID = 87.1>

  
carboxylic <EMI ID = 88.1>

  
oxo-6-hydroxy-4,5-dihydro-1,2,4-triazin-3-ylthio) methyl-3-cephem 4-carboxylique.

  
Another preferred group of cephalosporins of formula I is that represented by the following formula VI

  

 <EMI ID = 89.1>


  
These structures are eminently useful as intermediates in the preparation of other compounds according to the present invention. They are easily transformed into other compounds by

  
customary acylation methods in which the free amino substituent in position 7 is acylated with one of the other acyl substituents defined herein. Illustrative examples of those compounds represented by formula VI and presented in their free acid form are as follows:

  
 <EMI ID = 90.1> <EMI ID = 91.1>

  
The compounds of Formula I can be prepared by well known techniques for the preparation of cephalosporin. The. The products are readily available by displacement of an acetoxy function present in position 3 of a 3-acetoxymethyl compound of cephalosporin. Therefore, any of the

  
 <EMI ID = 92.1>

  
corresponding cephalosporins by the well-known displacement reaction. Therefore, a typical sequence of preparations yielding compounds of formula I from readily available 7-aminocephalosporanic acid (7-ACA) may be as follows:

  

 <EMI ID = 93.1>
 

  
 <EMI ID = 94.1>

  
 <EMI ID = 95.1>

  
7-ACA with the defined triazinylthio derivative. The resulting product is the corresponding 7-amino cephalosporin, a compound of formula I. This compound can be converted, by means of well known acylation techniques, into any of the defined 7-acylamidocephalosporins of formula I.

  
The preceding sequence can be modified such that the 7-ACA is first acylated in position 7 to give the corresponding 7-acylamido cephalosporanic acid or its aster. The resulting product, if it is an ester, is then dissociated to remove the C4 protective ester group, and the resulting acid is then subjected to the displacement reaction to obtain the compound of formula I. All the methods used to carry out these transformations are well known in the art.

  
The particular compound which is used to displace the acetoxy function in position 3 is a 3-mercapto-5-oxo-6-hydroxy4,5-dihydro-1,2,4-triazine unsubstituted in position 4, or substituted with a suitably in position 4 with a lower alkyl group as defined herein. These 3-mercapto1,2,4-triazines are prepared by the method described by Pesson et al., Bulletin de la Société Chimique de France, (1970), pages
1590-1599. According to the process described in this publication, 1,2,4-triazine is obtained by reaction of a thiosemicarbazide

  
with diethyl oxalate in the presence of sodium ethoxide. The reaction sequence is as follows:

  

 <EMI ID = 96.1>
 

  
 <EMI ID = 97.1>

  
lower alkyl group as defined herein. It should be noted that the product exists in tautomeric forms. The tautomeric aspect is detailed by Pesson et al., Bulletin de la Société

  
Chimique de France (1970), pages 1599-1606.

  
The displacement reaction mentioned above which is used in the preparation of compounds of formula I comprises the reaction of the corresponding 3-acetoxymethyl compound, preferably in a polar medium, with the 3-mercapto-1,2 derivative, Appropriate 4-triazine so that the acetoxy group is displaced.

  
The desired derivative of cephalosporin is then recovered. substituted in position 3.

  
This reaction can be carried out in a suitable manner by keeping the reactants in solution at a temperature such that,

  
 <EMI ID = 98.1>

  
until the desired derivative is obtained with optimum performance. The reagents are advantageously used in the ratio of about 1 molar equivalent of 3-acetoxymethyl cephalosporin to about 1 to about 10 molar equivalents of 3-mercapto-1,2,4-triazine. Preferably, the reaction is carried out using equivalent molar amounts of these reagents or a slight excess of the triazine compound. The pH of the reaction solution is advantageously maintained between about 5.0 and about 8.0.

  
It seems that the reaction occurs by a polar or ionic mechanism. It is therefore preferable to use

  
a polar medium for the reaction. This facilitates easy progress of the desired reaction. Water can be used as a polar medium and of course it is preferred. Any of the other typical polar solvents can also be used.

  
and well known. It is also possible to use as a maintenance medium for the reaction an excess of the displacement derivative of

  
 <EMI ID = 99.1>

  
The resulting reaction product can be separated from the reaction medium using a number of methods including crystallization, iontophoresis, paper chromatography and ion exchange resin chromatography.

  
The displacement reaction can be applied either to 7-amino acid or to 7-acylamino cephalosporanic acid. When using 7-ACA acid, the product can then be acylated <EMI ID = 100.1>

  
is a well known reaction and can be carried out by reacting cephalosporin with an acid halide or mixed anhydride representative of the desired acyl function. The particular acylation process is not important to the present invention.

  
The compounds of formula I are in themselves antibiotically active products and / or are synthetic intermediates of active antibiotic compounds. The compounds which exhibit more particularly an antibiotic activity are those in which R2 is hydrogen or an alkali metal cation. Compounds in which R 2 is different from hydrogen or of an alkali metal cation are readily converted into compounds containing them by cleavage techniques described above.

  
In the following table, the minimum inhibitory concentrations (MIC) in micrograms per milliliter (mcg / ml) of the compounds of formula I are given against a gram-negative organism, Escherichia coli, and against a gram-positive organism Staphylococcus aureus, resistant to penicillin. The MIC values are determined by the gradient plate technique

  
 <EMI ID = 101.1>

  
 <EMI ID = 102.1>

BOARD , ........

  
Antibiotic activity

  

 <EMI ID = 103.1>


  
Conc. inhibitory
 <EMI ID = 104.1>
  <EMI ID = 105.1>

BOARD

  
Antibiotic activity (continued)

  
Conc. inhibitory

  
minimal
 <EMI ID = 106.1>
  <EMI ID = 107.1>

  
Antibiotic activity (continued)

  
Conc. inhibitory

  
minimal
 <EMI ID = 108.1>
  <EMI ID = 109.1>

  
invention by the following examples.

  
Preparation of 3-mercapto-4-methyl-5-oxo-6-hydroxy-4,5-

  
dihydro-1,2,4-triazine

  
230 g of sodium are slowly added to a flask with a capacity of 22 liters containing 12.5 liters of anhydrous ethanol maintained under a nitrogen atmosphere. The mixture is kept at room temperature overnight, admitting a small but constant stream of nitrogen through the flask. The mixture is then heated to 50 ° C., and 1050 g (10 moles) of 4-methylthiosemicarbazide are added. The temperature of the mixture drops to 40 [deg.] C. Then 1530 g of diethyl oxalate was added to the mixture through a funnel. The addition is carried out at a speed sufficient to

  
 <EMI ID = 110.1>

  
 <EMI ID = 111.1>

  
The mixture is refluxed with stirring for 4 hours, during which time a precipitate forms. The precipitate changes appearance during the reflux period. By stopping the heating and the stirring, a white solid is gradually deposited in the bottom of the flask which leaves a clear pale yellow supernatant liquid. The mixture is left overnight. The mass of clear supernatant liquid is removed by suction and the crystalline product in the form of its sodium salt is collected from the residual mixture by filtration and then washed with dry ethanol.

  
The product is dissolved in water and the pH of the solution is adjusted to 5.5. It is filtered off and a small amount of the product is discarded. The pH of the filtrate is then adjusted to 1.5,

  
it is cooled and filtered to obtain 211.5 g of product,

  
 <EMI ID = 112.1>

Example 1

  
1.44 g of 3-mercapto are added to 36 ml of water with stirring.

  
 <EMI ID = 113.1>

  
 <EMI ID = 114.1>

  
and the pH is brought to 7.6 by addition of dilute acid. We add to

  
 <EMI ID = 115.1>

  
carboxylic acid, and the pH of the resulting mixture is adjusted to 6.9 while the cephem compound slowly dissolves. The mixture is stirred at 55 [deg.] C. After 1.75 hours, the pH of the mixture is <EMI ID = 116.1>

  
of dilute sodium hydroxide. After 17.5 hours the pH of the mixture is 6.1 and it is adjusted to 6.95 by addition of dilute sodium hydroxide. The mixture is heated in total for 21 hours.

  
The mixture is cooled in ice, filtered, and the filtrate is adjusted to pH 1.5. A solid precipitates. The solid was filtered, washed with dilute acid (pH 1.5), and air dried to afford 3.60 g of a light yellow powder. The powder is dissolved in 15 ml of a 2: 1 mixture of water and methanol at pH 6.5. The mixture is evaporated to a low volume (about 3 ml) on a rotary evaporator with the addition of isopropyl alcohol to prevent freezing until the end of evaporation on the rotary apparatus. The residual solution is placed in a column containing 50 g of Sephadex G-10 and eluted with water,

  
the first fraction being 5 ml and the following other fractions being approximately 15 ml. Fractions 5 to 8 are mixed, and the pH of the mixture is adjusted to 1.5 by adding dilute hydrochloric acid. The precipitated product was collected by filtration, washed with dilute acid (pH 1.5), and dried at

  
 <EMI ID = 117.1>

  
3-ylthio) methyl-3-cephem-4-carboxylic acid in the form of a light yellow powder.

  
 <EMI ID = 118.1>

  
2-CH2); 3.27 (s, 3H, N-CH3 triazine); and 1.36 ppm

  
 <EMI ID = 119.1>

Example 2

  
404 mg of the product of Example 1 are stirred in 4.5 ml of acetonitrile; however, we do not obtain a solubilization

  
 <EMI ID = 120.1>

  
after 5 minutes the solubilization is complete, and a solid begins to separate. After 1.5 hours, 0.5 ml of water is added, then the pH of the mixture is adjusted to 5.0 by addition of a saturated aqueous solution of ammonium carbonate. Most of the acetonitrile was removed by evaporation on the rotary apparatus, and isopropyl alcohol was added to the stirred residue until a slight precipitation appeared. We eliminate

  
the precipitate by filtration with a filter aid. There is a certain tendency for additional precipitation per passage <EMI ID = 121.1>

  
filtrate and diluted with isopropyl alcohol to obtain a further precipitate which is collected by filtration, washed with a mixture of isopropyl alcohol and water, and dried with water. air to obtain 0.18 g of a light yellow powder. This powder is mixed with 2.0 g of a similar product from a

  
 <EMI ID = 122.1>

  
 <EMI ID = 123.1>

  
30 ml of methanol. The pH of the mixture is slowly adjusted to 7.0 by

  
 <EMI ID = 124.1>

  
a small amount of insoluble product. Concentrate the filtrate to about 5 ml on a rotary evaporator, using isopropyl alcohol in the later stages to avoid freezing. The residual solution is placed in a column of Sephadex G-10 (28 g, 1.5 cm) in water, and 25 ml fractions are collected, water being used as eluent. Fractions 3 to 9 were combined. The pH of the combined fractions was adjusted to 3.5, and the product which precipitated was collected by filtration. The solid was washed with water which had been acidified to pH 3.6 by adding HCl, then dried to obtain 620 g of 7- (-amino) acid.

  
 <EMI ID = 125.1>

  
triazin-3-ylthio) methyl-3-cephem-4-carboxylic acid in the form of a light brown powder.

  
 <EMI ID = 126.1>

  
(s, 1H, C6H5-CH); 5.21 (d, 1H, 60 (-H); 4.43 (q,

  
 <EMI ID = 127.1>

  
(s, 3H, N-CH3 triazine).

Example 3

  
 <EMI ID = 128.1>

  
6-hydroxy-4,5-dihydro-1,2,4-triazine (15 mmol). The mixture, which has a pH of 2.8, is stirred, then the pH is adjusted to 7.25 by

  
 <EMI ID = 129.1>

  
 <EMI ID = 130.1>

  
carboxylic acid, and the pH of the resulting mixture is 3.2. The pH is carefully adjusted to 7.2 by adding 14.8 ml of hydroxide

  
 <EMI ID = 131.1>

  
4.25 hours, during which time the pH of the mixture increases to 5.35. The pH is raised to 7.17 by adding 0.7 ml of sodium hydroxide

  
 <EMI ID = 132.1>

  
during which the pH of the mixture becomes 5.99. We then adjust <EMI ID = 133.1>

  
 <EMI ID = 134.1>

  
the mixture in an ice bath, diluted to about 120 ml by adding water, and its pH is adjusted to 1.5 by addition, under

  
 <EMI ID = 135.1>

  
The resulting precipitate was filtered off, washed with dilute hydrochloric acid (pH 1.5), and air dried to obtain an off-white powder (5.28 g).

  
This product is stirred in 40 ml of water, during which time

  
 <EMI ID = 136.1>

  
concentrate the resulting solution by adding isopropyl alcohol and evaporation is carried out on a rotary apparatus until

  
that isopropyl alcohol has been removed. The final volume of the mixture is approximately 10 ml. The mixture is placed in a column of Sephadex G-10 (110 g, 2.5 cm, in water). We elect the column

  
with water and collected fractions of 15 to 16 ml each. Four fractions or mixtures of fractions are prepared, more particularly, fractions 5 and 6, fractions 7 and 8, fraction 9, and fractions 10 to 12. Each is stirred in the cold and the pH of each is adjusted to 1.5. by addition of hydrochloric acid

  
 <EMI ID = 137.1>

  
each precipitate by filtration, washed with dilute acid

  
(pH 1.5), then it is dried in air to obtain whitish powders (fractions 5 and 6, 1.26 g), (fractions 7 and 8:

  
0.84 g), (fraction 9: 0.31 g), and (fractions 10 to 12: 0.41 g). Thin layer chromatography (TLC) shows that each product is the same, and they are all combined to obtain 2.80 g of an off-white powder.

  
This product is dissolved in water, and the solution is acidified.

  
at pH 1.5. The precipitate is filtered off and air dried to give an off-white powder. The powder is dissolved in 25 ml

  
of tetrahydrofuran and isopropyl alcohol is added with stirring until a total volume of 200 ml is obtained. We

  
filter the mixture, 50 ml of isopropyl alcohol is added to the filtrate. The mixture is stirred and then filtered. The filtrate is evaporated to dryness and the residue is dissolved in water at pH 7.

  
The solution is acidified to pH 1.5, filtered and air dried.

  
 <EMI ID = 138.1>

  
as desired product.

  
 <EMI ID = 139.1>

  
2H, 3-CH2); and 3.31 (s, 3H, N-CH3 triazine).

Example 4

  
3.46 g of 7-formamido-3- acid are added to 20 ml of water.

  
 <EMI ID = 140.1>

  
(12.55 mmol). The resulting mixture is stirred and 1N sodium hydroxide is gradually added until the pH remains constant, 7.0. The mixture is then stirred at

  
 <EMI ID = 141.1>

  
to 20 ml and acidified to pH 1.2 by addition with cooling 3N hydrochloric acid. The resulting precipitate is filtered and immediately placed under a bell to dry in vacuo. 2.75 g of the dry product are ground in a mortar and then triturated 3 times each time with 150 ml of boiling isopropyl alcohol. The isopropyl alcohol solution is evaporated to dryness and the residue is triturated twice with 30 ml of ethyl acetate. The insoluble product is filtered off, washed with ethyl acetate and then dried to obtain 1.56 g of 7-formamido-3- (4-methyl-5-oxo-

  
 <EMI ID = 142.1>

  
4-carboxylic.

  
 <EMI ID = 143.1>

  
3.40 ppm (s, 3H, N-CH3 triazine).

Example 5

  
0.74 g of the product of Example 4 is stirred in 12 ml of dry methanol, and 1.5 ml of concentrated hydrochloric acid are added, during which time complete solubilization occurs. After a short time, a white solid begins to precipitate. Stirring is continued for 1.7 hours. and the mixture becomes cloudy with a white precipitate. The precipitate is filtered off and dried. This product is shown by TLC on 0.346 g of product to be an extremely pure sample of 7-amino-3- acid hydrochloride.

  
 <EMI ID = 144.1>

  
3.77 (b, 2H, 2-CH2); and 3.31 ppm (s, 3H, N-CH3 triazine).

  
 <EMI ID = 145.1>

  
To 36 ml of water is added 1.44 g (9 mmol) of 3-mercapto-4methyl-5-oxo-6-hydroxy-4,5-dihydro-1,2,4-triazine. The pH of the resulting mixture is 2.9 and is adjusted to 7.20 by the addition of

  
 <EMI ID = 146.1>

  
(9 mmol) of the sodium salt of 7- (2-thienyl) acetamido3-acetoxymethyl-3-cephem-4-carboxylic acid. The pH of the resulting mixture is 6.8 and is adjusted to 7.12 by the addition of 2 drops

  
 <EMI ID = 147.1>

  
hours, time during which the pH varies up to 6.03. We go back

  
 <EMI ID = 148.1>

  
 <EMI ID = 149.1>

  
during which the pH varies up to 6.02. We bring the pH to 7.09

  
 <EMI ID = 150.1>

  
heating for 1.5 hours (the total time at 55 [deg.] C being 20 hours). The resulting reaction mixture is then stirred in

  
 <EMI ID = 151.1>

  
 <EMI ID = 152.1>

  
filtration, washed with dilute acid (pH 1.5) and air dried to obtain a light yellow powder (3.51 g).

  
The solid is stirred in 25 ml of water, and slowly added
10 ml of methanol. The pH of the resulting mixture is adjusted to 6.8 by

  
 <EMI ID = 153.1>

  
an intermediate treatment with ultrasound on the resulting mixture. The resulting solution is concentrated to about 5 ml on a rotary evaporator, which allows the methanol to be removed from the mixture, then the concentrate is placed in a column of Sephadex G-10 (70 g, 2 cm column in water. ). The column is eluted with water, collecting fractions of 10 to 12 ml. Combine fractions 3 to 5, fractions 6 to 7, and fractions 8 to 10, and the resulting three fractions stirred cold and the pH adjusted to

  
 <EMI ID = 154.1>

  
the three resulting precipitates by filtration, washed with dilute hydrochloric acid (pH 1.5), and dried in air to obtain whitish powders, weighing respectively 2.13 g, 0.69 g and 0 , 17 g. Thin layer chromatographies on

  
each of the products indicate that they are virtually identical. Therefore, the products are combined and dissolved by stirring and boiling in a large volume of acetone (approximately
1200 ml), then the resulting solution is allowed to evaporate for <EMI ID = 155.1>

  
Crystalline 4-carboxylic acid.

  
NMR, DMSO d6: 9.1 (d, 1H, 7 &#65533; -NH): 7.36 & 6.95 (m, 3H, thienyl);

  
 <EMI ID = 156.1>

  
2H, 3-CH2); 3.79 (bs, 2H, 7-CH2); 3.68 (m, 2H, 2-CH2); and 3.33 ppm (s, 3H, N-CH3 triazine).

Example 7

  
To 20 ml of water is added 0.96 g of 3-mercapto-4-ethyl-5-oxo-6hydroxy-4,5-dihydro-1,2,4-triazine. The pH of the resulting mixture is 2.3 and is adjusted to 6.5 by the addition of 5 ml of sodium hydroxide.

  
 <EMI ID = 157.1>

  
7- (2-thienyl) acetamido-3-acetoxymethyl-3-cephem-4carboxylic acid. The pH of the resulting mixture is 5.7 and is adjusted to 7.2 by addition of 1N sodium hydroxide. The resulting mixture was stirred at 60 ° C. for a total of 18 hours, the final pH being 5.9. The mixture was cooled in ice and stirred, diluted to about 10 ml, and adjusted to pH 1.5 by addition.

  
 <EMI ID = 158.1>

  
filtration, washed with dilute HCl (pH 1.5), and air dried to obtain an off-white powder (2.01 g). We dissolve

  
the product at pH 6.8 in 20 ml of a 1: 1 mixture of water and methanol. The solution is evaporated on a rotary evaporator to a low volume (about 3 ml), and the residual solution is placed in a column of Sephadex G-10 (40 g, 1.6 cm column). The column is eluted with water, the first fraction being about 18 ml and the subsequent fractions about 5 ml. Fractions 4 to 11 were combined, and their pH was lowered to 1.5 by adding dilute hydrochloric acid to give 1.171 g of solid. The solid is dissolved in about 30 ml of a 3: 1 mixture of acetone and methanol. Air is blown over the surface of the solution to evaporate it. They are filtered and washed with acetone.

  
 <EMI ID = 159.1>

  
5-oxo-6-hydroxy-4,5-dihydro-1,2,4-triazin-3-ylthio) methyl-3-cephem4-carboxylique.

  
 <EMI ID = 160.1>

  
 <EMI ID = 161.1>

  
531 mg (3 mmol) of 3mercapto-4-methyl-5-oxo-6-hydroxy-4,5-dihydro-1,2,4-triazine are added to 12 ml of water with stirring. The

  
The pH of the resulting mixture was 2.3 and was adjusted to 6.5 by the addition of 2.9 ml of 1N sodium hydroxide. 1.54 g (3 mmol) are added

  
 <EMI ID = 162.1>

  
cephem-4-carboxylic acid with stirring and a mass is formed

  
 <EMI ID = 163.1>

  
solubilization and, while the mixture is maintained at this) temperature, the pH is adjusted to 7.0 and then the mixture is maintained at this value for 19 hours. After the first 16 hours the mixture is thick and gelatinous, so much so that stirring is stopped. The mixture is thoroughly mixed using a spatula, and heating is continued without stirring during the

  
 <EMI ID = 164.1>

  
the mixture is cooled in an ice bath and acidified to pH 1.5. The fluidity of the mixture is maintained by dilution at approximately
100 ml by addition of water. After stirring at pH 1.5 at room temperature for 1.5 hours, the mixture is filtered, washed with

  
 <EMI ID = 165.1>

  
cream colored. The powder is dissolved in tetrahydrofuran
(THF), filtered, and diluted with ethanol. The resulting solution is placed in a stream of air whereby a solid settles followed by a yellow gum. The gum is removed using a spatula. The resulting concentrate was filtered, and the product was washed with ethanol and air dried to give 861 m of an off-white solid. The recrystallization procedure is repeated

  
 <EMI ID = 166.1>

  
(4-methyl-5-oxo-6-hydroxy-4,5-dihydro-1,2,4-triazin-3-ylthio) methyl3-cephem-4-carboxylic acid in the form of an off-white powder.

  
NMR, DMSO d6-D20; 7.4 (bm, 4H, 2,5-dichlorophenyl); 5.70 (d, 1H,

  
 <EMI ID = 167.1>

  
3-CH2); 3.92 (b, 2H, 7-CH2); 3.69 (b, 2H, 2-CH2); and 3.33 (s, 3H, N-CH3 triazine).

Example 9

  
To 12 ml of water, 571 mg of 3-mercapto-

  
 <EMI ID = 168.1>

  
resulting mixture was 2.3 and adjusted to 6.6 by addition of 2.9 ml of 1N sodium hydroxide. 1.54 g (3 mmol) of

  
 <EMI ID = 169.1> <EMI ID = 170.1> cephem-4-carboxylic acid.

  
NMR, DMSO d6-D20: 7.4 (bm, 4H, 2,5-dichlorophenyl); 5.68 (d, 1H,

  
 <EMI ID = 171.1>

  
 <EMI ID = 172.1>

Example 10

  
350 mg (0.695 mmol) of acid are added to 12 ml of anhydrous THF

  
 <EMI ID = 173.1>

  
the addition of 917 mg (7 mmol) of N-trimethylsilylacetamide.

  
After dissolving, 1 ml of propylene oxide is added followed by

  
 <EMI ID = 174.1>

  
(chloroformyl) -methylamine dissolved in 2 ml of dry THF. The addition is carried out while maintaining the mixture at a temperature of -10 [deg.] C. The resulting mixture was stirred at -10 [deg.] C for 10 minutes then at room temperature for 15 minutes. Then 1 ml of water is added followed by the addition of 30 ml of an aqueous solution of sodium bicarbonate. The mixture is then cooled with

  
50 ml of a 6: 1 mixture of ethyl acetate and THF. We lower

  
 <EMI ID = 175.1>

  
50 ml of a 6: 1 mixture of fresh ethyl acetate and THF. The organic layer is separated and dried over magnesium sulfate. The mixture is then filtered and evaporated to dryness. The powdery residue is then dissolved in hot ethyl acetate and the ethyl acetate solution is concentrated until it becomes cloudy. Isopropyl alcohol is then added, and after completely redissolving, the mixture is again concentrated until the solution becomes cloudy. The concentrated solution is cooled overnight to obtain 234 mg

  
 <EMI ID = 176.1>

  
methyl-3-cephem-4-carboxylic.

  
NMR, DMSO d6-D20: 7.60 and 7.45 (two b, 9H, aromatic);

  
 <EMI ID = 177.1>

  
5.05 (d, 1H, 6 &#65533;-H); 3.66 (m, 2H, 2-CH2);

  
3.40 (s, 3H, N-CH3 triazine); and 3.17 ppm
(s, 3H, CON (CH3) CO) ..

Example 11

  
 <EMI ID = 178.1>

  
mix at room temperature for 2 hours. After dissolving, 1 ml of propylene oxide is added and the mixture is cooled.

  
 <EMI ID = 179.1>

  
 <EMI ID = 180.1>

  
in 5 ml of dry THF, and the reaction mixture is stirred for

  
 <EMI ID = 181.1>

  
then add 5 ml of water, and the THF is removed in vacuo. Then 50 ml of an aqueous solution of sodium bicarbonate are added to the residue. The resulting mixture is washed with ethyl acetate, and

  
 <EMI ID = 182.1>

  
The acidified aqueous mixture is then extracted with 100 ml of a 6: 1 mixture of ethyl acetate and THF. The organic layer is separated, dried over magnesium sulfate, filtered,

  
and the filtrate is evaporated to dryness. The residue obtained as a foam was dissolved in ethyl acetate, and ether was added to precipitate 140 mg of 7- - 3- (2-chlorobenzoyl) - acid.

  
 <EMI ID = 183.1>

  
which is collected by filtration.

  
NMR, DMSO d6-D20: 7.54 and 7.41 (two b, 9H, aromatic); 5.73

  
(d, 1H, 7 &#65533;-H); 5.59 (s, 1H, 7-CH); 4.07 (m, 2H, 3-CH2); 3.58 (m, 2H, 2-CH2); 3.28 (s, 3H, N-CH3 triazine); and 2.94 ppm (s, 3H, CON (CH3CO).

Example 12

  
350 mg (0.695 mmol) of acid are added to 20 ml of dry THF

  
 <EMI ID = 184.1>

  
the addition of 786 mg (6 mmol) of N-trimethylsilylacetamide. The resulting mixture is stirred at room temperature for 2 hours, after which time a solution is obtained. We then add
97.5 mg (0.71 mmol) of furoyl-2-isocyanate, and the reaction mixture was stirred for 1 hour at room temperature. 20 ml of water are then added to the solution. The volume of the mixture is reduced until cloudiness develops, after which time 40 ml of an aqueous solution of sodium bicarbonate are added. The <EMI ID = 185.1>

  
ethyl. The aqueous layer is separated, and the pH is reduced to 2.0

  
 <EMI ID = 186.1>

  
50 ml of ethyl acetate. The ethyl acetate extracts were combined, dried over magnesium sulfate, filtered and evaporated to give a residue as a foam. The residue is dissolved in hot ethanol, and the ethanolic solution is concentrated until cloudiness develops. The ethanolic solution thus concentrated is then cooled to

  
 <EMI ID = 187.1>

  
triazin-3-ylthio) methyl-3-cephem-4-carboxylic acid.

  
 <EMI ID = 188.1>

  
 <EMI ID = 189.1>

  
2-CH2); and 3.32 ppm (s, 3H, N-CH3 triazine). Example_13

  
A mixture of 381 mg (1 mmol) of 7- (1H-

  
 <EMI ID = 190.1>

  
200 mg (1.2 mmol) of 3-mercapto-4-methyl-5-oxo-6-hydroxy-4,5dihydro-1,2,4-triazine, and 1.25 mmol of sodium bicarbonate in 30 ml of buffer to pH 7. A small amount of solid remains insoluble in the mixture, the pH of the resulting mixture being about 6.6. The mixture is heated to about 64 [deg.] C. During the first hour, the pH of the mixture rises to 7.5 and then gradually decreases to about 6.7. Heating is continued for 5 more hours, after which time no further change in pH occurs. The mixture is then cooled, decanted with ethyl acetate, and the pH of the mixture is reduced to 2.7 by adding 20% hydrochloric acid. The organic layer is separated, then the aqueous layer is washed with additional ethyl acetate.

   Combine the original ethyl acetate layer and the washing ethyl acetate layer, and dry.

  
the complete mixture over magnesium sulfate, filtered, and evaporated to obtain 100 mg of 7- (1H-tetrazoleacetamido) -3- acid

  
 <EMI ID = 191.1>

  
Theory: C, 37.42; H, 3.14; N, 26.18 <EMI ID = 192.1>

Example 14

  
To 50 ml of dry tetrahydrofuran (THF), 1.85 g is added

  
(5 mmol) of 7-amino-3- (4-methyl-5-oxo-6-hydroxy-4,5-dihydro- acid

  
 <EMI ID = 193.1>

  
until a solution is obtained. The solution is then cooled to -20 [deg.] C.

  
Separately, 1.97 g (5 mmol) of the sodium salt of

  
 <EMI ID = 194.1>

  
glycine to a solution of 0.085 g (0.5 mmol) of N-trimethylsilylsuccinimide in 75 ml of dry tetrahydrofuran. 6 drops of N, N-dimethylbenzylamine are added to the resulting mixture. The resulting suspension is then cooled to -15 [deg.] C, and 0.52 g (5.5 mmol) of methyl chloroformate is added with stirring.

  
 <EMI ID = 195.1>

  
previous solution containing the cephalosporin core product. The resulting reaction mixture was stirred for 2 hours at -20 [deg.] C then for 1 hour at room temperature. High pressure liquid chromatography of the reaction mixture indicates the presence of approximately 60% of the desired product and 40% of the cephalosporin core starting material.

  
The reaction mixture is taken up by adding to the mixture of
10 ml of methanol. A precipitate forms which is removed by filtration. 10 ml of water are then added to the filtrate. The mixture is stirred for 15 minutes, and the resulting precipitate is filtered. The filtrate is evaporated in vacuo to about 40 ml, then the concentrated mixture is cooled overnight in a freezer. A precipitate forms which is collected by filtration.

  
The resulting collected solid is stirred in 15 ml of water. The pH of the mixture is adjusted to 1.1 by adding 3 drops of concentrated hydrochloric acid. The acidified mixture is then stirred for 5 minutes, and the insoluble products are filtered off. The pH of the filtrate is brought to 3.0 by the addition of sodium hydroxide, and the resulting mixture is stirred for 10 minutes at ice bath temperature. A precipitate forms which is collected by filtration. To the resulting filtrate is added 1 volume of isopropyl alcohol, and the mixture is evaporated in vacuo to about 10 ml. The concentrated solution was stirred in an ice bath for 15 minutes, and the resulting precipitate was collected by filtration, <EMI ID = 196.1>

  
cephem-4-carboxylic.

  
 <EMI ID = 197.1>

  
 <EMI ID = 198.1>

  
3.54 ppm (b, 5H, N-CH3 triazine and 2-CH2).

Example 15

  
252 mg (0.5 mmol) of acid are added to 25 ml of dry THF

  
 <EMI ID = 199.1>

  
650 mg (5 mmol) of N-trimethylsilylacetamide. The mixture is stirred for about 3 hours, at the end of which time a

  
 <EMI ID = 200.1>

  
 <EMI ID = 201.1>

  
the resulting mixture was stirred for 1 hour at room temperature, and the mixture was evaporated in vacuo to an oil. 20 ml of water was added to the oil, and the pH of the resulting mixture was brought to 8.0 by the addition of an aqueous solution of sodium bicarbonate. The resulting mixture was washed with ethyl acetate, and the pH of the aqueous layer was lowered to 2.0 by addition.

  
 <EMI ID = 202.1>

  
 <EMI ID = 203.1>

  
carboxylic.

  
NMR, DMSO d6-CD30D: 7.43 (b, 5H, CgHg); 5.76 (d, 1H, 7 "-H);

  
5.61 (s, 1H, C6H5-CH); 5.05 (d, 1H, 6 &#65533; -H) 4.16 (m, 2H, 3-CH2); 3.64 (b, 2H, 2-CH2);

  
 <EMI ID = 204.1>

  
 <EMI ID = 205.1>

Example 16

  
To 25 ml of dry THF is added 554 mg (1.1 mmol) of acid

  
 <EMI ID = 206.1>

  
for 3 hours until a solution is obtained. We

  
 <EMI ID = 207.1>

  
of 1-chloroformylimidazolin-2-one. The resulting mixture is stirred

  
 <EMI ID = 208.1>

  
ambient. 2 ml of water are then added to the mixture. We evaporate <EMI ID = 209.1>

  
25 ml of water in oil, and the pH is brought to 7.5 by adding sodium bicarbonate. The solution is washed with ethyl acetate, then the aqueous layer is acidified to pH 1.8 by adding acid.

  
 <EMI ID = 210.1>

  
with methanol, then the insolubles are filtered off with methanol. The resulting methanolic solution is slowly evaporated under vacuum until an oil is obtained. 25 ml of water are added to the oil, and the pH is brought to 7.5 by adding sodium bicarbonate. The solution is washed with ethyl acetate, then the aqueous layer is acidified to pH 1.8 by addition of IN hydrochloric acid. The resulting solid is filtered off and triturated with methanol, then the insolubles are filtered off with methanol. The resulting methanolic solution is slowly evaporated in vacuo until a precipitate is obtained. The precipitate is filtered off and dried to obtain

  
 <EMI ID = 211.1>

  
ppm (s, 3H, N-CH3 triazine).

Example 17

  
 <EMI ID = 212.1>

  
(5 mmol) of N-trimethylsilylacetamide. The mixture is stirred for about 3 hours, after which time a

  
 <EMI ID = 213.1>

  
 <EMI ID = 214.1>

  
2-one. The reaction mixture is stirred for about 30 minutes

  
 <EMI ID = 215.1>

  
add 2 ml of water to the mixture. The solution is then evaporated in vacuo until an oil is obtained. 25 ml of water are then added, and the pH of the mixture is increased to 7.5 by adding sodium bicarbonate. The solution is then washed with ethyl acetate, and the aqueous layer is separated and acidified to pH 1.8 by addition.

  
 <EMI ID = 216.1>

  
 <EMI ID = 217.1> <EMI ID = 218.1>

  
(b, 4H, N-CH2-CH2-N); and 3.37 ppm (b, 6H,

  
 <EMI ID = 219.1>

Example 18

  
 <EMI ID = 220.1>

  
(10 mmol) of hydroxybenzotriazole and 2.57 g (10 mmol) of acid <EMI ID = 221.1> <EMI ID = 222.1>

  
carbodiimide. The mixture was stirred while cooling in an ice bath for 2.75 hours. The mixture was then quickly filtered, and the solids washed with 10 ml of dry THF. The filtrate which has been collected is kept in an ice bath.

  
To 60 ml of dry THF containing 9.8 g (75 mmoles) of N-trimethylsilylacetamide, 3.73 g (10 mmoles) of 7-amino-3- acid are added.

  
 <EMI ID = 223.1>

  
approximately 1 hour, at the end of which almost complete solubilization occurs. The resulting solution was then stirred in an ice bath and after 15 minutes was added rapidly to the filtrate prepared above containing the acylating agent. The resulting mixture was stirred under cooling in an ice bath for 30 minutes and then at 32 [deg.] C for 3 hours. The resulting brown solution was quickly poured into ice water with stirring. Ether is added, and the pH of the aqueous phase is adjusted to 8.2. The phases are separated, and the aqueous layer is washed with an additional volume of ether. The aqueous phase is then subjected to evaporation in a rotary apparatus to remove residual ether. Ice is added to the resulting aqueous solution, and the mixture is quickly stirred while adjusting the pH.

  
 <EMI ID = 224.1>

  
resulting, and the solid was washed with dilute hydrochloric acid (pH 1.8), air dried, then vacuum dried.

  
 <EMI ID = 225.1>

  
pale yellow-brown.

  
 <EMI ID = 226.1>

  
 <EMI ID = 227.1> <EMI ID = 228.1>

  
N-CH3).

Example 19

  
1.60 g of the product of Example 18 is placed in a flask fitted with a stirrer. Gold then cools the flask in a bath

  
of ice-acetone. Then 60 ml of trifluoroacetic acid which has been cooled beforehand in an ice-acetone bath is quickly added to the flask. The resulting mixture is stirred for 15 minutes, during which time complete solubilization occurs. Thin layer chromatography (TLC) on the mixture shows that the reaction is complete. The reaction mixture is then subjected to evaporation in a rotary evaporator to obtain a gum. 60 ml of ethyl acetate are added to the gum, and the mixture is subjected to treatment with ultrasound, which gives a powder which is collected by filtration, and which is washed with acetate. ethyl, and air-dried to obtain 155 g

  
of a light brown product. A mixture of

  
75 ml of water and 10 ml of methanol. The resulting mixture is subjected

  
 <EMI ID = 229.1>

  
the mixture, and the pH of the filtrate is adjusted to 3.7. The resulting mixture was filtered again, and the solid which was collected was washed with dilute acid (pH 3.8) which was added to the filtrate. Isopropyl alcohol is then added to the filtrate and the total is evaporated in a rotary apparatus until reduction to a small volume. Additional isopropyl alcohol is added to the residue, and the mixture is filtered. The solid collected is washed with a 1: 1 mixture of water and isopropyl alcohol maintained at pH 3.8. The resulting solid is dried to

  
 <EMI ID = 230.1>

  
methyl-5-oxo-6-hydroxy-4,5-dihydro-1,2,4-triazin-3-ylthio) methyl3-cephem-4-carboxylique.

  
 <EMI ID = 231.1>

  
(d, 1H, 7 &#65533;-H); 5.38 (s, 1H, 7-CH); 4.99

  
(d, 1H, 6 "-H); and 3.28 ppm (bs, 3H, N-CH3 triazine).

Example 20

  
To 24 ml of dry tetrahydrofuran, 408 mg (0.8 mmol) of the product of Example 19, and 919 mg (7 mmol) of N-trimethylsilylacetamide are added. The solubilization is complete after 10 minutes, and after 15 minutes, the solution is placed in an <EMI ID = 232.1>

  
remove the resulting mixture from the ice bath and hold it

  
 <EMI ID = 233.1>

  
low volume (about 10 ml) and ice water is added. The resulting suspension is then stirred, and the pH is adjusted to 6.5. The resulting solution is washed with 2 volumes of ether, then the aqueous phase is subjected to evaporation in a rotary apparatus until the ether has been removed. The aqueous mixture is then acidified to pH 1.7, and the resulting solid is collected by filtration and partially dried on the filter. The wet product is then dried under vacuum to obtain 96 mg of acid.

  
 <EMI ID = 234.1>

  
3- (4-methyl-5-oxo-6-hydroxy-4,5-dihydro-1,2,4-triazin-3-ylthio) methyl-3-cephem-4-carboxylic acid in the form of a yellow-brown powder blade.

  
 <EMI ID = 235.1>

Example 21

  
Using the same process as that described in Example 20, but replacing N-chloroformyl-N, N'-dimethylurea with 217 mg

  
 <EMI ID = 236.1>

  
hydroxy-4,5-dihydro-1,2,4-triazin-3-ylthio) methyl-3-cephem-4carboxylic.

  
NMR, DMSO d6: 9.52 (d, 1H, NH); 8.72 (d, 1H, NH); 7.45, 7.10 and

  
6.99 (m, 3H, thienyl); 5.88 (d, 1H, 7-CH);

  
5.74 (q, 1H, 7 &#65533;-H); 5.08 (d, 1H, 6 &#65533;-H); 4.08

  
(m, 2H, 3-CH2); 3.78 (b, 4H, N-CH2-CH2-N); 3.60
(m, 2H, 2-CH3); 3.3'4 and 3.28 ppm (two s, 6H

  
 <EMI ID = 237.1>

Example 22

  
To 15 ml of dry THF is added 317 mg (0.5 mmol) of the salt of

  
 <EMI ID = 238.1>

  
 <EMI ID = 239.1>

  
resulting 0.59 g of N-trimethylsilylacetamide. The mixture is stirred for 2 hours at room temperature, after which time solubilization occurs. The

  
 <EMI ID = 240.1>

  
of propylene oxide. 123 mg (0.5

  
 <EMI ID = 241.1>

  
The resulting solution then stirred at room temperature for 1.5 hours, after which time the reaction mixture was filtered. 1 ml of water is added to the filtrate; however, no precipitate occurs. The mixture is then evaporated under vacuum at approximately

  
10 ml, and thereto was added 50 ml of ethyl acetate followed by the addition of 50 ml of water. The pH of the mixture is brought to 7.5 by adding sodium bicarbonate. Add to the aqueous layer
50 ml of fresh ethyl acetate and 15 ml of THF, and the pH of the aqueous layer is lowered to 2.5 by addition of hydrochloric acid

  
 <EMI ID = 242.1>

  
dried over magnesium sulfate, and filtered. The filtrate is then evaporated in vacuo to about 10 ml, then 20 ml of ether is added. The mixture is then filtered to obtain 150 mg of acid.

  
 <EMI ID = 243.1>

  
3-ylthio) methyl-3-cephem-4-carboxylic acid.

  
NMR, DMSO d6: 7.35 (b, 4H, o-chlorobenzoyl); 6.96 (q, 4H,

  
p-hydroxyphenyl); 5.83 (q, 1H, 7 &#65533;-H); 5.47

  
 <EMI ID = 244.1>

Example 23

  
To 25 ml of dry THF containing 1.18 g (9 mmol) of N-trimethylsilylacetamide, 634 mg (1.0 mmol) of the trifluoro salt is added.

  
 <EMI ID = 245.1>

  
3-cephem-4-carboxyliqua. Solubilization occurs 15 minutes after the end of addition of the cephalosporin compound. We

  
 <EMI ID = 246.1>

  
ice-acetone, and 84 mg (1 mmol) of sodium bicarbonate is added followed by the addition of 1 ml of propylene oxide. 226 mg (1 mmol) of 1-chloro- <EMI ID = 247.1> are then added to the resulting mixture.

  
reaction mixture at room temperature and stirred at room temperature for 1.5 hours. The reaction mixture is then filtered, and 1 ml of water is added to the filtrate. He ... not

  
no precipitation occurs. Then evaporated under vacuum

  
the mixture to about 10 ml, and 100 ml of a 6: 1 mixture of ethyl acetate and THF are added along with 50 ml of water. The pH of the mixture is brought to 7.0 by adding sodium bicarbonate. The aqueous layer is then separated from the organic layer, and the pH of the aqueous layer is lowered to 2.0 by addition of 1N hydrochloric acid. The mixture is then filtered and the solid collected is washed with isopropyl alcohol and dried to obtain

  
 <EMI ID = 248.1>

  
(q, 2H, 3-CH2); 4.08 (b, 4H, N-CH2-CH2-N); 3.7-3.5
(b, 5H, 2-CH2 and N-CH3 triazine); and 3.44 ppm (s, 3H, SO2CH3).

Example 24

  
Using the method of Example 22 on a 1 mmol scale, the cephalosporin is reacted with 151 mg (1 mmol) of N-chloroformyl-N, N'-dimethylurea in the presence of sodium bicarbonate at

  
the place of the triethylamine used in Example 22, for

  
 <EMI ID = 249.1>

  
dihydro-1,2,4-triazin-3-ylthio) methyl-3-cephem-4 -carboxylique. NMR, DMSO d6-D20: 7.08 (q, 4H, p-hydroxyphenyl); 5.72 (d, 1H,

  
 <EMI ID = 250.1>

Example 25

  
Using the method of Example 24, but using

  
 <EMI ID = 251.1>

  
 <EMI ID = 252.1>

  
 <EMI ID = 253.1>


    

Claims (1)

<EMI ID = 254.1> <EMI ID = 255.1> <EMI ID = 256.1> <EMI ID = 257.1> inferior ; R &#65533; is hydrogen, an alkali metal cation, or a group giving an easily removable ester; and R is 0 hydrogen or the R'-C- group in which R 'is: hydro- <EMI ID = 258.1> <EMI ID = 259.1> <EMI ID = 260.1> <EMI ID = 261.1> nitrobenzyl, benzyl, 2,2,2-trichloroethyl, t-butyl, or p-methoxybenzyl and A 'is ur. C2-C4 alkanoyl, C2-C4 haloalkanoyl, benzoyl, halobenzoyl, 2,4-dinitrophenyl, or phtaLoyl group; or R 'is a group of formula <EMI ID = 262.1> <EMI ID = 263.1> halogen, hydroxyl, or aminomethyl; Z is 0 or S; and m is 0 or 1; <EMI ID = 264.1> <EMI ID = 265.1> <EMI ID = 266.1> 1-tetrazyl, or a substituted phenyl group of the formula <EMI ID = 267.1> formula wherein a and a 'are as defined above; and Q is the hydroxyl, formyloxy, acetoxy, carboxyl, sulfo, amino, or -NHY group in which Y is the benzyloxycarbonyl group, <EMI ID = 268.1> <EMI ID = 269.1> <EMI ID = 270.1> you 0 <EMI ID = 271.1> <EMI ID = 272.1> mono- or diphenylamino, or R "'and V taken together form <EMI ID = 273.1> <EMI ID = 274.1> <EMI ID = 275.1> <EMI ID = 276.1> R "-CH2 formula wherein R" is 2-thienyl; 3-thienyl; 2-furyl; 2-oxazyl; 2-thiazyl; 1-tetrazyl; benzo- <EMI ID = 277.1> triazolyl; or 1- (3-cyano) -1,2,4-triazolyl. 2. A compound according to claim 1, which is called the acid. <EMI ID = 278.1> <EMI ID = 279.1> dihydro-1,2,4-triazin-3-ylthio) methyl-3-cephem-4-carboxylique. 6. A compound according to claim 1 which is called the acid. <EMI ID = 280.1> 3-ylthio) methyl-3-cephem-4-carboxylic acid. 7. A compound according to claim 1 which is called the acid. <EMI ID = 281.1> dihydro-1,2,4-triazin-3-ylthio) methyl-3-cephem-4-carboxylic acid. 9. A compound according to claim 1 which is called the acid <EMI ID = 282.1> 3-cephem-4-carboxylic. 11. A compound according to claim 1 which is called the acid. <EMI ID = 283.1> 3-cephem-4-carboxylic. 12. A compound according to claim 1 which is called the acid. <EMI ID = 284.1> carboxylic. 13. A compound according to claim 1 which is called the acid. <EMI ID = 285.1> 3-cephem-4-carboxylic. 15. A compound according to claim 1 which is called the acid. <EMI ID = 286.1> <EMI ID = 287.1> 3-ylthio) methyl-3-cephem-4-carboxylic acid. 16. A compound according to claim 1 which is called the acid. <EMI ID = 288.1> methyl-3-cephem-4-carboxylic. 17. A compound according to claim 1 which is called the acid <EMI ID = 289.1> triazin-3-ylthio) methyl-3-cephem-4-carboxylic acid. 18. A compound according to claim 1 which is called the acid <EMI ID = 290.1> 3-ylthio) methyl-3-cephem-4-carboxylic acid. 19. A compound according to claim 1 which is called the acid. <EMI ID = 291.1> 1,2,4-triazin-3-ylthio) methyl-3-cephem-4-carboxylic acid. <EMI ID = 292.1> <EMI ID = 293.1> 3-ylthio) methyl-3-cephem-4-carboxylic acid. 21. A compound according to claim 1 which is called the acid. <EMI ID = 294.1> 3-ylthio) methyl-3-cephem-4-carboxylic acid. 22. A compound according to claim 1 which is called the acid <EMI ID = 295.1> 1,2,4-triazin-3-ylthio) methyl-3-cephem-4-carboxylic acid. 23. A compound according to claim 1 which is called the acid <EMI ID = 296.1> 1,2,4-triazin-3-ylthio) methyl-3-cephem-4-carboxylic acid. 24. A compound according to claim 1 which is called the acid <EMI ID = 297.1> 1,2,4-triazin-3-ylthio) methyl-3-cephem-4-carboxylic acid. 25. A compound according to claim 1 which is called the acid <EMI ID = 298.1> 26. A process for the preparation of a cephalosporin compound of <EMI ID = 299.1> <EMI ID = 300.1> <EMI ID = 301.1> with a triazinylthio derivative of formula <EMI ID = 302.1> optionally the acylation of the compound thus obtained in which R is hydrogen, or Q is the amino group; and, if desired, removing the protecting group from the amino group and / or the carboxyl group. 27. Process for the preparation of cephalosporin compounds from <EMI ID = 303.1> claim 1, essentially as described above with particular reference to the examples. 28. Cephalosporin compounds of formula I, wherein <EMI ID = 304.1> as defined above with particular reference to the examples.