DE1670119A1 - Non-anaphylactic cephalosporins and processes for their preparation - Google Patents

Description

¹¹ Non-anaphylactic cephalosporins and process for their preparation "

Priority: July 31, 1965, Great Britain, No .: 32 857/65

The invention relates to non-anaphylactic cephalosporins and a process for their preparation.

Patent application B 77 957 IVd / 12p already describes a process for the production of non-anaphylactic benzylpenicillin has been described, the high molecular weight, strongly antigenic substance contained in the penicillin by purification processes Will get removed.

The allergic symptoms that occur when penicillin is administered to certain people are well known, and one of the dangers of penicillin therapy is that it can cause severe reactions in susceptible individuals can cause death by shock.

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It is known to have the same effects when administered of cephalosporins occur in certain people can.

It is an object of the invention to provide non-anaphylactic cephalosporins that do not cause any reactions when administered, i.e. they are likely to have a lower one Awareness potential.

The non-anaphylactic cephalosporins according to the invention correspond to the general formula:

RCO. NH. CH- CH CH ₉

Ii I ² (D

CO N ^ JZ - CH ₂ A

COOM

in which RCO is an acyl group, A is a lower alkanoyloxy, Benzoyloxy, hydroxyl or pyridinium group or together with M a simple carbon-oxygen bond and M. hydrogen, a non-toxic, pharmaceutically acceptable cation, has an anionic charge when A is a pyridinium group, or together with A denotes a simple carbon-oxygen bond.

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The erfinaun ^ sgecäase process for the manufacture of these substances is characterized in that the cephalosporins or those used for their production and obtained by fermentation Starting products are subjected to one or more purification stages in order to reduce the high molecular weight, strongly antigens contained therein Remove substance.

The cephalosporin is produced either by fermentation or directly obtained by converting a starting ephalosporin which has itself been obtained by a fermentation process is. The removal of the high molecular weight substance can be from the original fermentation product or one of its Derivatives take place regardless of whether this is a starting product or the end product.

When the substituent A in the given formula is a pyridinium group represents, the cationic charge on this group is determined by the anionic charge on the carboxyl group

BATH

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balanced, so that the molecule has the structure of a zwitterion and M thus means an anionic charge. The hydroxyl group which A may represent, the carboxyl group of the heterocyclic ring may further terephthalate tert intramolecularly duroh be, wherein a Laotonring is formed, according to the indication representing that A and 11 together form a simp _o he carbon-oxygen bond. The invention also includes the non-toxic, pharmaceutically acceptable salts, where H is a cation. Suitable cations are alkali metal and alkaline earth metal ions, the ammonium ion and organic amine cations, e.g. lower alkylammonium groups,

The cephalosporins can for example by acylation of 7-aminooephalosporanic acid or one of its suitable derivatives, depending according to the type of substituents A and M, with a functional derivative of the acid RCOOH or by reacting the acid itself can be obtained with the 7-amino compound. Both reactions are carried out in a manner known per se. The 7-aminocephalosporanic acid can by acid hydrolysis from Cephalospor ■ can be produced in C. By treating cephalosporin C with the tertiary base pyridine before acid hydrolysis, an inner salt of the 3rd pyridinium methyl-7-aminodeoephalosporanic acid is formed. These acids themselves can be purified, or they are made from purified cephalosporin O. produced, or after the acylation, the acyl compounds are purified. In all cases, cleaning the

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high molecular weight, strongly antigenic substance removed.

The high molecular weight, strongly antigenic substance is likely to exist essentially from protein or polysaooharide compounds, to which the groups derived from cephalosporin are covalently bound * These groups form during fermentation by reaction of the cephalosporin with the protein or polysacoharide compound in the medium.

The preferred purification methods are based on the molecular weight differences, for example when using molecular sieve material or in dialysis, other suitable methods are differential adsorption and electrophoresis _{or the like}

A method which is carried out with a modified dextran as molecular sieve material is particularly suitable. This dextran is obtained by fermentation of sugar, 'whereby the linear macromolecules of the dextran become three-dimensional Network of polysaccharide chains are linked o Sin suitable material is commercially available under the. Came "Sephadex" available.

Test with commercially available cephaloridin (formula I, in which H is a 2-thienylmethyl group, Λ is a pyridinium group and M

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16 7 O i 19 - 6 -

mean an anionic charge) have the presence of a showed high molecular weight fraction which reacts with an antiserum directed against the fenioiloyl determinant.

The invention is illustrated by the following examples,

example 1

An aqueous solution of 20 g. Commercially available cephaloridin is placed in a sack of a Visking dialysis tube and dialyzed extensively for 5 days with distilled water. The residue, which has largely been freed from cephaloridin, is concentrated by evaporation and freeze-dried, until 66 days of a solid substance is obtained (a high percentage of this amount of substance is probably residual cephaloridin or its breakdown products). A solution of 2 mg of this solid substance in 0.5 ml of water causes passive anaphylactic skin ■ ...

reaction emerged in guinea pigs that were rioted against the penioilloyl determinant by intradermal injections Rabbit antibodies have been sensitized. The Penicilloyle Specificity is further confirmed by inhibiting the reaction with N-penioilloyl-f-aminocaproic acid.

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Table I.

Diameter (in am) of the blue coloration in the passive anaphylactic skin reaction

Evocative
A _n t L / 50 Antiserum dilutions 2 mg cephaloridin
residue * 26th 1/250 1/1250 1/6250 2 mg cephaloridin
arrears + IO ng
N ~ penicilloyl- £ -
aminocaproic acid - 21 + w «M mm

* That means the residue of less than 0.5 g.

Example 2 '_' '. ■

A solution of 5 g of commercial cephaloridin in 25 ml of water is treated in a column of 6.35 µm in diameter and 91.44 cm in length made of Sephadex G 10. The material is eluted with water flowing through it at a rate of 1 liter / hour. A graphic representation of the elution behavior shows that the cephaloridin, which has strong ultraviolet absorption and is recognizable through the hydroxylamine sample because of the β-lactam ring, was eluted from the column with a peak activity at 2600 ml.

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The ultraviolet absorption at 254 nyu, however, shows a pronounced maximum value (peak) at 1200 ml (the void volume the column). This indicates the presence of a substance contained in the origin * High Cephaloridin, which is due to their molecular weight has been excluded from the Sephadex G 10 molecular sieve material. This faction shows one clear yellow color. A second, smaller peak is also found at 1600 ml. The test of the first harps After the combined fractions have been concentrated to 10 ml, the tip of the high molecular weight substance shows that 1.0 ml (corresponding to 0.5 g cephaloridin) are capable of passive anaphyiactosis To evoke an eal reaction,

Table II

Diameter (in mm) of the blue coloration in the passive anaphylactic skin reaction »

Evoking antigen

1.0 ml high molecular weight Xon concentrate from Sephades Q 10

Anti-serum dilutions

1/100

16

1/500

3/2500

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The experiments described in Examples 1 and 2 show that the commercially available cephaloridin contains a high molecular weight substance which can be separated off by dialysis or gel filtration with Sephadex G 10. This high molecular weight Substance reacts with antibodies of the penicilloyl determinant. The cephaloridin exiting through the dialysis membrane and also the fractions containing cephaloridin from the Sephadex O 10- * Columns are recovered, evaporated and 'freeze-dried, whereby one obtains products / which are practically free of high molecular antigenic impurities «'

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Claims (5)

Patent claims 1. Non-anaphylactic cephalosporins of general formula N. RCO. NH. CH CH CH CO NC CH ₉ A (D "COOM. and their non-toxic salts, where RCO is an acyl group means A is a lower alkanoyloxy, benzoyloxy, hydroxyl or pyridinium group or together with M a simple carbon-oxygen bond and M represents hydrogen, a non-toxic, pharmaceutically acceptable cation, a anionic charge when A is a pyridinium group, or together with A a simple carbon-oxygen bond means. 2. Non-anaphylactic cephaloridine (R = 2-thienylmethyl group, A = pyridinium group, H = anionic charge). 3. A process for producing non-anaphylactic cephalosporins according to claim 1, formula I, characterized ge -kennze rest t that the cephalosporins or the starting products used for their preparation and erhalte- NEN by fermentation are subjected to one or more purification steps contained therein, high molecular weight to remove strong antigenic substance. 009849/1944 4. The method according to claim 3, characterized in that that the high molecular substance is separated by a method based on molecular weight difference will. 5. The method according to claim 4, characterized in that that the high molecular weight substance is removed by a molecular sieve material or by dialysis, 6 «The method according to claim 3,« characterized in that that the high molecular substance is removed by differential adsorption or electrophoresis 009849/1944