DE1445831C3 - 7-Phenylacetamido-cephalosporanic acid derivatives, processes for their preparation and pharmaceutical formulations - Google Patents

Description

// X

CH ₇ -CO-NH-CH

^ -CH ₂ OCOCH ₃

wherein R _{1 has} the meaning given in claim 1, or a water-soluble salt of this compound at a pH of 5 to 8 in water or a mixture of water and a water-miscible organic solvent with a compound of the general formula

CO ₂ H

lating agent, which is different from a compound of the general formula

y ν

35

wherein R _{2 has} the meaning given in claim 1, or that one converts the compound of the formula in a manner known per se

40

CH, -COOH

derives and, if desired, a compound of the general formula I obtained in this way, in which R _{1 is} a nitro group, treated with hydrogen in the presence of palladium-carbon.

4. A pharmaceutical formulation consisting of a compound according to claim 1 and usual auxiliary and / or carrier materials!

H, N-CH

O = C

CH, OCOCH,

45

CO, H

with a compound of the general formula

The invention relates to new derivatives of 7-aminocephalosporanic acid (7-ACA) i.e. H. a compound whose structure is usually represented as follows will:

wherein R _{2 has} the meaning given above, and reacts the resulting compound of the general formula

55

H ₂ N-CH

O = C N

60

CH, -]

NH ₂ CH-CHe 2CH ₂
O = C-Ns 3C-CH ₂ OCOCH ₃

CO ₂ H

65 A derivative of 7-ACA, which has a valuable antibiotic activity, in particular against gram-positive _{organisms wherein R 2 has} the meaning given in claim 1, and reacts with a common phenylacetyl attack by penicillinase-producing bacteria

is stable, is N- (7-Phenylacetamidoceph-3-em-3-ylmethyl) -pyridinium-4-carboxylate dar:

QH ₅ • CH ₂ -CO-NH

this compound I a in addition to its resistance to attack by penicillinase producing staphylococci generally have considerably improved activity against both important have gram-positive organisms as well as against gram-negative organisms. Some of these derivatives are also more resistant to attack by penicillinase from staphylococci than compound I a is.

The invention relates to new compounds of the following

It has now been found that certain derivatives obey the general formula

- ^ V-CH ₂ -CO-NH-CH- ^{/ S} "

CH ₂ -N

(II)

COI

wherein R _{1 is} a hydrogen atom, a lower-alkoxycarbonyloxy, hydroxy, amino, nitro or lower-alkanoyloxy group and R _{2 is} a carbamoyl, carboxy or lower-alkoxycarbonyl radical in the 3- or 4-position of the pyridinium ring.

The compounds of this type generally have beneficial activity against gram negative organisms.

The compounds in which R ₂ denotes a carbamoyl group are of particular interest.

Under the terms "lower alkanoyloxy" and "lower alkoxy" as used herein are To understand groups with 1 to 4 carbon atoms, preferably with 1 or 2 carbon atoms.

The invention also relates to a process for the preparation of the compounds of the general formula II, which is characterized in that a compound of the general formula is used in a manner known per se

/ V

-CH, -CO-NH-CH

CH, OCOCH,

(III)

CO, H

wherein R _{1 has} the meaning given above and the resulting compound is omnidirectional, or a water-soluble salt of this compound
at a pH of 5 to 8 in water or a
Mixture of water and a water-miscible organic solvent with a compound of the general formula

my formula H, N - CH

45

= C N

(IV)

H, N-CH

= C N

CH ₂ OCOCH ₃

CO ₂ H

with a compound of the general formula

wherein R _{2 has} the meaning given above,

in which R _{2 has} the meaning given above, or that the compound of the formula is converted in a manner known per se

wherein R _{2 has} the meaning given above, with a conventional phenylacetylating agent which is different from a compound of the general formula

55

CH ₇ COOH

derives, reacts and, if desired, a compound of the general formula II obtained in this way, in which R _{1 is} a nitro group, treated with hydrogen in the presence of palladium-on-carbon.

The preferred method according to the invention consists in using the compound of the general formula III to implement with the nucleophilic agent of the general formula IV given above. If Salts of the compounds of the general formula III are used, suitable salts are, for example the sodium, potassium or ammonium salts. In those cases where the nucleophilic agent is in

If water is not very soluble, the aqueous medium may become miscible with water organic solvent such as Ν, Ν-dimethylformamide or acetone, added, although the yield may occasionally be reduced.

The reaction temperature is usually at least 15 ⁰ C, but is generally lower than 70 ° C.

The required reaction time depends on the temperature used, but can be changed by a previous experiment.

The reactants are advantageously used in a ratio of about 1 molar equivalent of the Compound III applied to 1 to 10 molar equivalents of the nucleophilic agent IV. The pH the solution is preferably maintained at 6-7. If necessary, the pH of the solution should . to the desired value by adding a buffering agent, for example ammonium acetate, or when using an alkali salt of the compound corresponding to the general formula III Addition of, for example, acetic acid can be adjusted.

The reaction product can be obtained from the reaction mixture, which is, for example, unreacted compound of the general formula III or other substances, by customary methods, such as Crystallization, electrophoresis, paper chromatography or by treatment with an ion exchange resin, be separated.

A convenient method is to use an excess of the compound of the general formula IV is reacted with the compound of general formula III in water until the optimum yield the desired compound II is obtained. The excess of compound IV and some unreacted Compound III are then with an organic solvent, for example methylene chloride, extracted and the remaining solution through an anion exchange resin, for example in the acetate form, given to all compounds with free carboxyl groups including the remainder to remove the unreacted compound III. The resulting solution is then concentrated, for example in a rotary evaporator, and freeze-dried. The residue can then be taken from a suitable Solvent, for example methanol, recrystallized or by precipitation, for example from a methanolic solution by adding acetone.

If the nucleophilic agent IV contains a carboxyl group, for example in the case of nicotinic and isonicotinic acids, it is desirable to add a base such as ammonium hydroxide to the reaction medium in order to ensure complete dissolution and a favorable pH adjustment. In the case of ammonium hydroxide, after passing through an anion exchange resin column, in acetate form, the ammonium ions are obtained in the form of ammonium acetate, which then contaminates the product. In order to free the product from ammonium acetate, the product can be ^{treated with a cation exchange resin in the H +} form. At this time, while the products are not adsorbed on an anion exchange resin, they are adsorbed on the cation exchange resins at low pH values due to the protonation of the carboxyl group and its presence as cations. When the resin is subsequently treated with a base, such as ammonia, with careful pH adjustment (pH 7), the products are converted back to their original zwitterionic form and displaced from the resin by the ammonium ions.

The compounds corresponding to the general formula III given above can by Reaction of 7-ACA with a conventional phenylacetylating agent, conventional phenylacetylating agent are in general those as they are also used for the phenylacetylation of aminoamides, for example peptides, are suitable, and these include in particular phenylacetyl halides, in particular the chloride or bromide, and mixed anhydrides, derived for example from a phenylacetic acid and derive an alkyl haloformate.

The acylation is conveniently carried out in an aqueous solution Medium, for example an aqueous, water-miscible ketone, such as acetone, or in aqueous Tetrahydrofuran, preferably also in the presence of an acid binder, for example sodium bicarbonate, accomplished. The pH is preferably kept between 5 and 7 during the reaction, which can be carried out at temperatures between 0 and 25 ° C. The acylation can also be carried out in an organic solvent as the medium, for example ethyl acetate, where z. B. is simply heated to reflux.

On the other hand, these compounds can be obtained according to the invention by first adding 7-ACA the selected nucleophilic agent of the general formula IV to a compound of the general formula V is reacted, whereupon the compound obtained is then acylated with a phenylacetylating agent will. These reactions can generally be carried out under the above for the nucleophilic and acylation reactions, when carried out in the preferred order, the conditions described be performed. , ■ · ..-., In addition, the connections can be made accordingly of the general formula V from a compound of the general formula

HOOC • CH (CH ₂ J ₃ CONH-CH

NH ₂

CH ₂ N

(VI)

wherein R _{2 has} the meanings given above by employing the methods known for the conversion of cephalosporin C to 7-ACA. The compound corresponding to the general formula VI can be obtained from cephalosporin C by reaction with nucleophilic compounds

of the general formula IV are prepared using corresponding processes as described above for the Reaction of compounds of general formula III with compounds of general formula IV should be applied.

The quaternary ammonium compounds specified here are made with reference to the

CH ₂ -CO-

Substance cepham (see J.A. C. S, 1962, 84, 3400).

A particularly important compound obtainable according to the invention is N- (7-Phenylacetamidoceph - 3 - em - 3 - ylmethyl) - 3 '- carbamoylpyridinium-4-carboxylate are:

This compound shows marked biological activity in vivo against Staph. aureus, E. coli, B. proteus and Salmonella sp. It is also active against penicillin-sensitive strains of Stap. aureus and active against penicillin-resistant strains of the same organism. It is also shown against staphylococcal penicillinase extremely resistant. The compound also maintains high serum levels in vivo. Another benefit of this connection is the greater stability in vivo than is the case with compound I a.

Other compounds of particular interest according to the invention are compounds of the general type formula

// V

CH ₂ -CO-NH

CH, -

CONH,

wherein R _{1 is} an amino group, a hydroxyl group, a lower-alkoxycarbonyloxy group, especially the methoxycarbonyloxy group, or lower-alkanoyloxy group, especially the acetoxy group, the carbamoyl group being in the 3- or 4-position of the pyridinium ring.

The compounds obtainable according to the invention can be administered in human or veterinary medicine in any usual manner as other antibiotic substances, e.g. B. penicillin and neomycin, be formulated. Such masses can in the usual way with the help of the necessary pharmaceutical Carriers or absorption means are represented.

Thus, the compounds can be used for injection preparations both in solution and in suspension Media, e.g. B. sterile, pyrogen-free water, or as dry preparations that are used for the instant Manufacture of injectable preparations are suitable to be manufactured. The connections can continue in the form of preparations for topical use, e.g. B. lotions, ointments or creams are produced, 'which are formulated with suitable carriers for such preparations. The crowds can also be in shape tablets or capsules or liquids for oral administration.

For veterinary medicine, the compounds can be adjusted in the manner customary in veterinary medicine especially for injection as animal cereals.

In general, the. doses used in human medicine in adults in the range from 200mg per dose upwards, administered e.g. four times a day.

The following examples serve to further illustrate the invention. The ultraviolet absorption values were obtained from solutions in water or aqueous phosphate buffer at pH 6.0.

The paper chromatography in Examples 1 to 4 was buffered with 0.1 N sodium acetate (pH 5) Paper carried out. The solvent system used was with the volume in brackets are set, ethyl acetate (8), n-butanol (1), 0.1 n-sodium acetate (8); it was run downhill. In the other examples, the paper chromatography was also carried out on the same paper carried out, either buffered to pH 6 using a solvent system of n-butanol (4), water (5), ethanol (1) or buffered to pH 5 using ethyl acetate as solvent.

The electrophoresis was carried out on papers with a gradient of 10 to 30 volts per centimeter. Buffer solutions were (parts by volume in brackets) (a) pH 1.9, acetic acid (84), formic acid (17), acetone. (105), water (495); (b) pH 7, 0.05 n-disodium hydrogen phosphate, the pH with phosphoric acid was discontinued.

The fractions were shown as dark spots on the paper chromatograms and electrophoretograms found when the papers were irradiated with ultraviolet light.

example 1

N- (7-PhenylacetamidoCeph-3-em-3-ylmethyl) -S'-carbamoyl-pyridinium ^ -carboxylate

5 g of 7-phenylacetamidocephalosporanic acid were added with stirring in 55 ml of water containing 10.3 g of nicotinamide (6.6 equivalents) dissolved. The solution obtained of pH 4.0 was held under nitrogen at 35 ° C for 65 hours and then was extracted with four times 20 ml of methylene chloride, giving some 7-phenylacetamidocephalosporanic acid was removed, but the nicotinamide was not removed

609 515/490

60

would. The aqueous solution was then poured down a column of anion exchange resin in the acetate form, and the eluates containing the nicotinamide derivative, judged polarimetrically, were pooled and freeze-dried. The freeze-dried solid was first stirred with four 100 ml portions of acetone to dissolve the nicotinamide, and the residue was then stirred with about 20 ml of methanol, some of which dissolved and separated from an insoluble, gummy solid. When this methanol solution was added to a large volume of acetone (about 500 ml), the nicotinamide derivative precipitated out (494.5 mg), A _max 262n ^ (El * _m 236). When rubbed with water, the gummy solid turned into an almost white solid, which was the desired nicotinamide derivative (307.2 mg), Z ₁₁₁₃₁ 261 ηΐμ (Ej? _M 293), [α] Ϊ + 157 ° (c 0.57 ) R _PACA / p _y 1.0.

Analysis for C ₂₂ H ₂₀ N ₄ O ₅ S ■ 2H ₂ O:

Calculated ... C 54.1, H 4.95, N 11.5, S 6.6%; found .... C 53.9, H 4.8, N 11.5, S 6.7%.

On freeze-drying, the filtrate gave a further amount of the desired derivative (268.7 mg), A _max 261 πΐμ (EJ * 266). i _S

Example 2

N- (7-Phenylacetamidoceph-3-em-3-ylmethyl) -3'-carboxypyridinium-4-carboxylate

30th

7.5 N ammonium hydroxide was added with stirring to a suspension of 3.46 g of nicotinic acid (about 2.5 equivalents) in 40 ml of water until the acid had dissolved, a solution with a pH of 6, 2 was created. 5 g of 7-phenylacetamidocephalosporanic acid was added, and the pH of the mixture was adjusted from 4.9 to 6.35 by further addition of 7.5 n-ammonium hydroxide. The solution was kept under nitrogen at 37 ^° C. for 64 hours and then extracted with 100 ml of methylene chloride. The aqueous phase was concentrated to about 24 ml in a rotary evaporator at 40 ⁰ C and was then added to a column of an anion exchange resin in the acetate form (25 χ 2.5 cm). The eluates containing the desired derivative, which was estimated polarimetrically, were pooled and freeze-dried. The solid obtained in this way was redissolved in about 10 ml of water and this solution was stirred with strongly acidic cation exchange resin in the hydrogen form at a pH of 3.2. More resin was added until the pH was 1.85, whereupon the mixture was filtered and the resin washed with water. The resin was then suspended in water and stirred with a magnet while the pH was adjusted from 2.4 to 7.1 by adding 7.5N ammonium hydroxide. After the resin had been removed by filtration, the filtrate was freeze-dried and gave the desired nicotinic acid derivative (0.616g, 9.5%), k _max 260πΐμ (E {* „255). This material was stirred with 50 ml of dry acetone, filtered off and dried (553 mg), / _max 261 πΐμ (EU 258), [«] ₀ + 16 ° (c 1.0 m 40% aqueous dimethylformamide), R _PACA / _Py 0.0.

Analysis for C ₂₂ H ₂₁ N ₃ O ₆ S · 4H ₂ O: ^{6 $}

Calculated ... C 50.0, H 5.5, N 8.0, S 6.1%;
found .... C 50.1, H 5.2, N 9.5, S 7.2%.

Similarly, N- (7-Phcnylacetamidoceph - 3 - em - 3 - ylmethyl) - 4 '- carbamoylpyridiniuin-4-carboxylate from 7-phenylacetamidocephalosporanic acid and isonicotinic acid amide (Example 3) and N - (7 - Phenylacetamidoceph - 3 - em - 3 - ylmethyl) -4'-carboxypyridinium-4-carboxylate from 7-phenylacetamidocephalosporanic acid and isonicotinic acid (Example 4) produced. The following results were obtained:

at base % Out Km _x Π 1 "ü M ° game prey No. (last Step) (ma) 3 Isonicotine 10 261 288 + 3O ⁰¹ ) acid amide 4th Isonicotine 9 260 - 258 + 15 ⁰² ) acid 261

') 50% aqueous dioxane.
² ) 40% aqueous DMF.

B ei s ρ i e1 3

N- (7-Phenylacetamidoceph-3-em-3-ylmethyl) -4'-carbamoylpyridinium-4-carboxylate

Isonicotinic acid derivative:
Analysis for C ₂₂ H ₂₀ N ₄ O ₅ S · H ₂ O:

Calculated ... C 56.2, H 4.7, N 11.9, S 6.8%;
found .... C 56.7, H 5.0, N 12.0, S 5.5%.

Other sample:
Analysis for C ₂₂ H ₂₀ N ₄ O ₅ S ■ 2H ₂ O:

Calculated ... C 54.1, H 5.0, N 11.5, S 6.6%;
found .... C 54.6, H 5.0, N 11.7, S 7.0%.

Example 4

N- (7-Phenylacetamidoceph-3-em-3-ylmethyl) -4'-carboxypyridinium-4-carboxylate

Isonicotinic acid derivative:
Analysis for C ₂₂ H ₂ IN ₃ O ₆ S 4H ₂ O:

Calculated ... C 50.0, H 5.5, N 8.0, S 6.1%;
found .... C 49.6, H 5.2, N 9.5, S 7.5%.

Example

N- [7- (p-Hydroxyphenylacetamido) -3-em-3-ylmethyl] -4'-carbamoylpyridinium-4-carboxylate

a) p-Hydroxyphenylacetic acid was in the Tetrahydropyranyläther, F, = 98 to 100 ° C, converted in 78% yield via the.-Methyl ester. 3.56 g this ether was dissolved in 230 ml of dioxane and 2.04 ml of triethylamine were added, and to the well-cooled 1.40 ml of ethyl chloroformate were added to the solution. After 30 minutes a A solution of 4.1 g of 7-ACA in 16 ml of water and 2.04 ml of triethylamine was added over the course of 10 minutes and the solution was brought to room temperature over the course of 2 hours. After implementation was completed 800 ml of water were added and the suspension was washed twice with 100 ml of diethyl ether, with a gave a clear solution, which was adjusted to pH 2.5 and extracted three times with 200 ml of ethyl acetate. the collected organic extract was washed with water, dried and distilled, whereby

about 50 ml resulted, which after dilution with 500 ml petroleum ether 3.77 g of crude 7- (p-hydroxyphenylacetamido) - cephalosporanic acid tetrahydropyranyl ether resulted.

3.63 g of the crude oily derivative were dissolved in 40 ml of ethanol, 30 ml of water and 0.70 ml of hydrochloric acid and left to stand at room temperature for 3 hours; the ethanol was removed under reduced pressure at 25 ^° C., the suspension was diluted with 50 ml of water and the product was extracted three times with 50 ml of ethyl acetate. After removal of the solvent, 2.06 g (35%) of 7- (p-hydroxyphenylacetamidocephalosporanic acid remained in the form of a colorless, hygroscopic glass.

λ 227 πΐμ max.Ej * = 268 1 0.005% ige ' ⁵

λ 262 ηΐμ max. Ej * = 176 [methanol
λ 269 ΐημ max.Ej * _m = 174 j solution

Rf (butanol system) = 0.55.

20th

b) 1.50 g of 7- (p-hydroxyphenylacetamido) cephalosporanic acid, 0.9 g Isonikotinamid and 10 ml of water were reacted together C for 16 hours at 5O ^0; the suspension obtained was diluted with 30 ml of water and washed with 40 ml of dichloromethane. The aqueous solution was placed in a column of an anion exchange resin (acetate form) and the column was washed out with water; 200 ml of the eluate were collected and concentrated to 5 ml at 25 ° C. under reduced pressure. After dilution with 400 ml of acetone, 0.21 g (12.1%) of N - [7 - (p - hydroxyphenylacetamido) - ceph - 3 - em-3 - ylmethyl] - 4 '- carbomoylpyridinium - 4 - carboxylate were obtained in the form a very finely divided white solid.

λ 260 ηΐμ max.Ej * _m = 228 λ water
λ 266 ηΐμ max.Ej * = 285 J 0.0025%
Rf (ethyl acetate system) = 0.00.

Example 6

N- [7- (p-methoxycarbonyloxyphenylacetamido) -

ceph-S-em-S-ylmethyrj ^ '- carbamoylpyridinium-

4-carboxylate

a) p-Methoxycarbonyloxyphenylacetic acid was after I. N. Gorgache νa et al., Zhur. Obshchei. Khim, 27, 2276-82 (1957) and converted into the acid chloride transferred by refluxing with thionyl chloride. 5.85 g of the acid chloride were condensed with 6.0 g of 7-ACA in aqueous acetone in the presence of an excess of sodium bicarbonate, 10.0 g (98%) of 7- (p-methoxycarbonyloxyphenylacetamido) cephalosporanic acid resulted.

λ 263 ηΐμ max.Ej * = 182 (methanol 0.004%).
R _f (butanol system) = 0.81.

b) 4.64 g of 7- (p-Methoxycarbonyloxyphenylacetamido) cephalosporanic acid were dissolved in 70 ml of water and 2.42 g Isonikotinsäureamid at 50 ⁰ C and .. ,,. the solution at 5O ⁰ C for 20 hours implemented ". The resulting mixture was washed twice with 50 ml dichloromethane and added to aqueous liquids over a column of an anion exchange resin (acetate form). eluate and washings were concentrated to about 5 ml under reduced pressure and with Acetone diluted to 500 ml, whereby

35

40

45 gave 1.588 g (30.2%) of N- [7- (p-methoxycarbonyloxypheny lacetamido) - ceph - 3 - em - 3 - ylmethyl] - 4 '- carbamoylpyridinium-4-carboxylate.

λ 264 ηΐμ max.Ej * _m = 229 (water solution
0.003%) = 0.00.

R _f (ethyl acetate system)

B ei s ρ i e 1 7

N- [7- (p-acetoxyphenylacetamido) -ceph-3-em-3-yimethyl] -4'-carbamoylpyridinium-4-carboxylate

a) 4.0 g of p-acetoxyphenylacetic acid were converted into the acid chloride with thionyl chloride and reacted with 5.0 g of 7-ACA in aqueous acetone in the presence of excess sodium bicarbonate. 7.0 g (84.5%) of 7- (p-acetoxyphenylacetamidocephalosporanic acid became a white crystalline solid receive.

λ 260 ηΐμ max.Ej * = 190 (sodium salt
in water 0.003%).

Rf (butanol system) = 0.63.

b) 3.0 g of 7- (p-acetoxyphenylacetamido) -cephalosporanic acid were mixed with 1.64 g of isonicotinamide reacted as in Example 5, whereby 0.60 g (17.5%) N - [7 - (p - acetoxyphenylacetamido) - ceph - 3 - em-3 - ylmethyl] - 4 '- carbamoylpyridinium - 4 - carboxylate.

/ 263 ηΐμ max. Ej * „= 233 (aqueous solution
0.0025%).

R _f (ethyl acetate system) = 0.00.

Example 8

N- [7- (p-Nitrophenylacetamido) -ceph-3-em-3-ylmethyl] -4'-carbamoylpyridinium-4-carboxylate

a) p-nitrophenylacetyl chloride was made from 6.0 g of p-nitrophenylacetic acid produced and used to convert 7.5 g of 7-ACA into 10.3 g. (85%) 7- (p-Nitrophenylacetamido) -cephalosporanic acid used.

b) 6.6 g of 7- (p-nitrophenylacetamido) -cephalosporanic acid were mixed with 3.36 g of isonicotinamide such as implemented in Example 5, whereby 0.80 g (10.6%) of the isonicotinamide derivative resulted.

λ 265 ΐημ max.Ej * = 385,
λ 268 m μ max.Ej * = 389.
Rf (ethyl acetate system) = 0.00.

55

Example 9

N- [7- (p-Aminophenylacetamido) -ceph-3-em-3-ylmethyl] -4'-carbamoylpyridinium-4-carboxylate

0.640 g of N - [7 - (p - nitrophenylacetamido) - ceph-3 - em - 3 - ylmethyl] - 4 '- carbamoylpyridinium - 4 - carboxylate were dissolved in 80 ml of warm water; the solution was cooled and placed in a pressure vessel; 0.640 g of palladium on carbon (5%) were added and the suspension was shaken _{under hydrogen at a pressure of 4.2 atmospheres for 2 hours.} The catalyst was filtered off and the aqueous solution freeze-dried, 0.200 g (26.6%) N - [7 - (p - aminophenylacetamido) - ceph - 3 - em-

3 - ylmethyl] -4 '- carbamoylpyridinium - 4 - carboxylate.

λ 242 ΐτίμ max. E | * _m = 242 \ (water solution λ 263 ηΐμ inflexion E \ J _a = 224 I 0.002%)
Rf (ethyl acetate system) = 0.00.

Example 10

N- (7-Phenylacetamidoceph-3 _r en-3-ylmethyl) -4'-methoxycarbonylpyridinium-4-carboxylate

7-Phenylacetamidocephalosporanic acid (3.3 g), 2.3 g of methyl isonicotinate and 30 ml of water were mixed well and kept at 50 ^° C. for 16 hours, after which the reaction mixture was cooled and diluted to 50 ml. The suspension was extracted three times with 20 ml of methylene chloride and poured into a small column of an anion exchange resin in the acetate form. The column was washed with water and 200 ml of eluate was collected, concentrated to 3 ml under reduced pressure and poured into 300 ml of acetone. The finely divided precipitate was collected, washed with acetone and diethyl ether and dried in a desiccator. Yield: 0.136 g, 3.4%.

λ 263 m μ max.Ej * _m = 250 (aqueous solution
0.002%).

Rf (ethyl acetate system) = 0.00.

shaken for a long time, the catalyst was filtered off and the filtrate was concentrated to 5 ml under reduced pressure. When added to 400 ml of acetone a white precipitate resulted which was collected with acetone and diethyl ether washed and dried in a desiccator. Yield 0.450g (36.3%).

/ 242 m μ max.Ej * _m = 270,

/ 258 mμ inf. Ej * "= 246.

Rf (ethyl acetate system) = 0.00.

Example 13

N- [7- (p-Hydroxyphenylacetamido) -ceph-3-em-3-ylmethyl] -3'-carbamoylpyridinium-4-carboxylate

6.5 g of 7 - (p - hydroxyphenylacetamido) - cephalosporanic acid were mixed with 3.87 g of nicotinic acid amide in 40 ml of water reacted at 50 ° C. for 17 hours. After cooling, the suspension was extracted twice with 30 ml of methylene chloride and on a Column of an anion exchange resin cast in the acetate form. The column was filled with water washed, the eluate concentrated to 5 ml under reduced pressure and added 5 ml of methanol; this Solution was poured into 500 ml of acetone and the white precipitate collected, well with acetone and Washed diethyl ether and dried in a desiccator. Yield 2.0g (26.7%).

λ 264 ΐημ max.Ej * _m = 190 (water 0.002%).
Rf (ethyl acetate system) = 0.00.

Example 11

N- [7- (p -nitrophenylacetamido) -ceph-3-em-3-ylmethyl] -3'-carbamoylpyridinium-4-carboxylate

20.0 g of 7 - (p - Nitrophenylacetamido) - cephalosporanic acid were added to a solution of 11.2 g of nicotinic acid amide in 200 ml of water and the suspension is kept at 50 ⁰ C for 18 hours. After cooling, the suspension was washed twice with 100 ml of methylene chloride, separated from a rubbery deposit and poured onto a column of an anion exchange resin in the acetate form. The column was washed with water, and the combined eluates were concentrated to 5 ml under reduced pressure and poured into 400 ml of ethanol. The precipitate was separated off, washed with acetone and diethyl ether and dried in a desiccator. Yield: 1.45 g (6.35%).

λ 266 max.EJ * = 368 (aqueous solution
0.002%).

R _f (ethyl acetate system) = 0.00.

Example 12

N- [7- (p-aminophenylacetamido) -ceph-3-em- • S-ylmethyVJ ^ -carbamoylpyridinium ^ -carboxylate

1.32 g of N - [7 - (p - nitrophenylacetamido) - ceph-3 - em - 3 - ylmethyl] - 3 '- carbamoylpyridinium - 4 - carboxylate were dissolved in 40 ml of water and 1.32 g of palladium-carbon (5.0% Pd.) were added. The suspension was under hydrogen at 4.2 atmospheres for 4 hours at spi e 1 14

N- [7- (p-acetoxyphenylacetamido) -ceph-3-em-3-ylmetnyl] -3'-carbamoylpyridinium-4-carboxylate

The above compound was made in exactly the same manner as in Example 13. yield 1.56 g (34%).

A264n ^ max. Ej * = 219

(0.002% in 50% aqueous methanol).
⁴ SR _f (ethyl acetate system) = 0.00.

The biological properties of the compounds prepared according to the examples are as follows Table summarized. The Staph. Aureus strains A and C were penicillin resistant, while strain B was penicillin sensitive.

The compounds T ₁ , T ₂ and T ₃ , the measured values of which are given in the bottom three columns of the table below, belong to the prior art. "Compound T ₁ " means 7-thienyl-2'-acetamidocephalosporanic acid (Journal of the American Chemical Society, 84, 3402, Table 1, second compound), "Compound T ₂ " means 7-phenylacetamidocephalosporanic acid (Journal of the American Chemical Society, 84, 3402, Table 1, penultimate compound), and "Compound T ₃ " means the 7-phenylacetamidocephalosporanic acid-C _A compound, which is taken from "The Biochemical Journal", Vol. 79, 1961, p. 414, is known. The table below shows that the products obtainable according to the invention are superior to the compounds belonging to the prior art in at least one of the tests carried out.

Example no. Pipe thinning test (; Staph. • / ml) gram negative S. typhi- Pr. Vulgaris Protection of mouse aureus E. coli munum (EDjo / mg / kg / dose) gram positive Strain B subcutaneous administration Staph. 0.08 Staph. 31 31 S. aureus E. coli aureus 0.3 aureus 31 8th 31 Strain B Strain A 0.16 Strain C 62.5 62 250 1 1.25 0.16 62 8th 16 2.5 10 2 2.5 0.16 31 16 31 6.25 50 3 0.62 0.3 16 62 62 1.5 25th 4th 2.5 0.6 31 8th 31 3 40 5 b) - 0.02 8th 31 125 250 2.5 10 6 b) - 0.3 8th 125 16 62 3 35 7 b) 0.6 0.16 2 16 4th - 10 35 8 b) 0.6 0.04 4th 31 - 250 - - 9 - 0.3 8th 31 - 31 3 12.5 10 0.16 0.3 6.25 16 - 31 <1.5 15th 11th 1.25 0.15 0.5 33 - 31 3 > 50 12th 0.6 0.16 4th 16 16 8th <1.5 20th 13th 1.25 0.25 1.0 125 16 16 3 35 14th 1.25 3.1 1.0 62 125 1.5 20th Connection T ₁ 1.25 1.25 62 > 25 > 50 Connection T ₂ 1.6 0.8 > 25 > 50 Connection T ₃ 3.1 3.1 1.5 15th

Claims (3)

Patent claims: 1. T-phenylacetamido-cephalosporanic acid derivatives of the general formula CH, - CO - NH - CH CH, -N wherein R _{1 is} a hydrogen atom, a lower-alkoxycarbonyloxy, hydroxy, amino, nitro or lower-alkanoyloxy group and R _{2 is} a carbamoyl, carboxy or lower-alkoxycarbonyl radical in the 3- or 4-position of the pyridinium ring. 2. N- (7-Phenylacetamidoceph-3-em-3-ylmethyl) -3'-carbamoylpyridinium-4-carboxylate. 3. Process for the preparation of 7-phenylacetamido-cephalosporanic acid derivatives according to claim 1, characterized in that a compound of the general formula is used in a manner known per se