PROCEDURE FOR THE PRODUCTION OF CEPHALOSPORANSAEUREDERIVATEN.

The invention concerns a procedure for the production of a 7-Aminocephalosporansäuredeñvates of the formula HOOC (CH2) 3 - CONHì (3 " CH2 X C OObí III where X hydrogen, hydroxyl or Acetoxy and M hydrogen or X Pyñdinio and M a negative load mean ®, by the fact characterized that one Cephalosporin C or a Deñvat of the same the formula converted into the connection of the formula III wird.

2. Procedure according to requirement 1, by the fact characterized that the reaction mixture treats to decomposition of surplus hydrogen peroxide with a Thiosulfat wird.

3. Procedure according to requirement 1, by it characterized that in the ct-Ketoderivat of the formula II RA is hydrogen and R carboxyl, Aroyl, in particular Benzoyl or Carbamoyl and that formula II in particular Glyoxylsäure, Brenztraubensäure, Phenylglyoxal, A-Ketophenylessigsäure, Glyoxylsäureamid or - Keto n butancarbonsäure means, ss 4th procedures according to requirement 1, by the fact characterized that in the et-Ketodeñvat of the formula II g 1 carboxyl and R2 Phenyl ist.

Procedure according to requirement 1, by the fact characterized that the ct-Ketoderivat of the formula uses II or its salt in Ibis 50fach molecular portion of the Cephalosporins C or its derivative of the formula I or its salt wird.

6. Procedure according to requirement 1, by it characterized that one accomplishes the conversion at a pH value from 3,0 to 8.0, it 7th procedure according to requirement 1, by the fact characterized that one the Umsetmmg in presence of a more zweioder trivalent metal ion, in particular of copper, manganese, Kal “OOC-CH (CH2) 3 - CONH I I ÷ G00 so or a salt of it converts R with a A-Ketoderivat of the formula: C-R2 II IL O where g 1 carboxyl, Aroyl or Carbamoyl is, if RA is hydrogen, and/or g 1 carboxyl is, if RA is alkyl or Aiyl, or with a salt of it, in presence of Wasserstoffperoxià or with additional treatment with hydrogen peroxide, whereby the n (et-Ketoadipoyl), developed as intermediate product - 7-nminocephalosporansäure of the formula IV C OOM IV converted into the connection of the Formel/li wird.

A goal of the invention is a commercially usable Herstellnngsveffahren, which works with low costs and with good yield and to Deñvaten of the 7-Aminocephalosporansäure leads which, those for their part output products to s the synthesis of antibiotics of the type of the Cephalospoñne with niedñger toxicity and broad pharmakologischem range of application darstellen.

When manufacturing processes for 7-Aminocephalosporansäurederivate of the formula III are such well-known, with those the J0 CephalosporinverbindungI, received by fermentation, are umgcwandclt into the 7-Aminocephalosporansäurederivat III, using D-Aminosäureoxidase (Belgian patent 736,934, Japanese patent Kokai (publication) No. 39595/72 and the Japanese patent No. 7158/75). With Js these procedures a large portion of C becomes - Ketoadipoyl-7aminocephalosporansäure or Deñvaten of it the formula IV HOOC jí (CH2) 3-CONH {--[-- S " O 0 “CH2 X COOH as intermediate product in an educated manner. In the following this connection shortened as t - Ketoadipoyl-7-ACA-Derivat designates. To the Inhibiernng of the education of this t-Ketoadipoyl-7-ACA-Deñvates IV, Natñumazid or similar enzyme inhibitors in the reaction system was suggested too verwenden.

The procedures mentioned exhibit meanwhile different Nac teiÂe. So Iür the commercial production enormous Ansrüstungen are necessary, for production of the DAminosäureoxidase. Furthermore the Nata'iumazid or ähnas a licher enzyme inhibitor, which must be used in large quantities, is gef'åhrlich for the human body. And finally the production of a sturdy final product is connected with difficulties, so that all in all the procedures mentioned no commercially favorable production erlauben.

Due to the aforementioned disadvantages the inventors undertook expanded studies for the location of a completely new procedure, which belongs not to the vorgenaunten Kategore, and found thereby that the 7-Aminocephalosporan4s will receive acid derivatives with high yield can by treatment of the Cephalosporinverbindung with one iKetoderivat, in presence of Wasserstoffperoxid.

During the production of the e-Ketoadipoyl-7-ACA-Derivates by conversion of the Cep alospoñnverbindung with the C - in such a way Ketoderivat develops besides anch a 7-Aminocephalosporansäuredeñvat. Meanwhile a-Ketoadipoyl-7-ACADeñvat is chemically extreme as intermediate product instabil.

In accordance with the procedure according to invention the developed intermediate product transferred in a stable 7-Aminoces5 phalosporansäurederivat by treatment with hydrogen peroxide, whereby itself the 7-Aminocephalosporansäuredeñvat selectively and in good yield bildet.

The conversion for the formation of the ct-Ketoadipoyl-7-ACADerivates from the Cephalospoñnverbindung is accomplished, by one the latter preferably in aqueous solution with the ct-KetoderJvat for reaction bringt.

Preferential ct-Ketoderivate for the procedure according to invention is Glyoxylsäure, Brenztraubensäure, Phenylglyoxal, Glyoxylsäureamid, t-Ketophenylessigsäure and Natriumsc ct-keto-n-butanoat. Under these the Glyoxylsäure is suitable best for the production of 7-Aminocephalosporansäure with high yields. Favourably the reaction 621,124 durchgefülårt at a pH within the range of 3,0 to 8.0, in particular from 3,5 to 6,0. If the pH value in the reaction mixture exceeds 8.0, the Cephalosporinverbindung and the ot-Ketoadipoyl-7-ACA-Derivat will decompose unstably and itself by secondary reactions. With the result that the yield at desired a-Ketoadipoyl-7-ACA-Deñvat or at the derivative of the 7-Aminocephalosporansäure become unwanted low. The portion of the ct-Ketoderivat is stöchiometñschäquimolar with the assigned Cephalosporinverbindung, however in an appropriate way is appropriately at least increased, according to the reaction conditions, e.g. the kind of the Cephalosporinverbindung of the Reakfionstemperatur and the kind and quantity of the existing impurities, so that on a mol of the Cephalospoñnverbindung i to 50 mole at Ketoderivat can find use. An unnecessarily large Übersc uss is not economic against it although he no special influence on the reaction ausübt.

As particularly favourable, the conversion speed of the Cephalosporinverbindung with a-Ketoderivat proved dnrc to addition of a metal salt, as e.g. from copper, manganese, calcium, aluminum, iron, nickel or cobalt, as well as an acid, like sulfuric acid, acetic acid, to increase Propionsäure or hydrochloric acid. The Metallsaiz preferably used in a molecular portion from I/t0 to 20 times of the molecular quantity of the assigned Cephalospoñnverbindung. Wishing case can be used an organic base together or in place of the metal salt mentioned, in order to shorten the response time. In this case the training is inhibiert by Nebertreaktionen and it arises better results. Preferential ones organic bases are tertiary cyclische amines, like Pyñdin, secondary cydisehe amines, like Imidazol or aliphatic amines, like tri ethyl amine, Dimethylamin or Methylarnin. The portion of organic base depends thereby with advantage after their kind as well as on the reaction conditions, i to 30 times the molecular portion of Cephalosporinverbindung amounts to however favourable. Unnecessarily large surplus of organic bases makes the cleaning more difficult of the desired 7-Aminocephalosporansäurederivates and is not therefore not erwünscht.

The procedure according to invention favourably accomplished at a temperature within the range of 0 to 50°C. Since meanwhile the Cephalosporinverbindung used as output products and the ct-Ketoadipoyl-7-ACA-Deñvat, which will receive as intermediate product, are chemically unstable, the observance of not unnecessarily high temperatures is recommended. Best the reaction accomplished below room temperature. Further the reaction duration of the procedure according to invention affected by different Reaktionsbedingnngen, e.g. by present or absence of the metal salt or the organic base, by the hydrogen ion concentration and by the reaction temperature. If no metal salt or no organic base present is, the response time usually lies between 3 and 24 hours, against what with presence of a metal salt or an organic base the response time within the range of 5 minutes until 4 hours lie kann.

Metal set organic base response time yield * (hours) (of %) no no 5.0 56.4 no 2.0 77.1 no 2.0 77.3 1.0 92.0 • yield at 7-Aminocephalosporansäurederivat.

As mentions above, a-Ketoadipoyl-7-ACA-Deñvat is chemically extremely instable and can be also converted 621,124 into a 7-Aminocephalosporansäurederivat by treatment with acid or by extension of the response time. Meanwhile the et-Ketoadipoyl-7-ACA-Derivat disappears not completely from the reaction liquid and its separation is quite schwieñg. If the 7-Aminocephalospos ransäurederivat are therefore manufactured soli, “- the Ketoadipoyl-7-ACA-Deñvat in the Reaktiouslösung converted according to invention with Wasserstoffperoxid.

To the Einbringnng of the hydrogen peroxide into the reaction mixture it and/or its aqueous solution can do added werlo that or it can a connection be used, the hydrogen peroxide in the Reaktioussystem forms, like e.g. Natfiumperborat.

The necessary quantity of hydrogen peroxide depends on the Reaktiousbedingungen, e.g. on the pH value of the Reakas fionsgemisches, the Reakfionstemperatur as well as on kind and concentration of the Cephalospoñnverbindung, which were used as Ausgangsmateñal, however preferably lies within the range of 1 to 30 times the molecular portion to the used Cephalosporinverbindung. As Ausgangsmatefial zo the used Cephalosporinverbindung and the received 7Aminocephalosporansäurederivat are easily oxidized, under decomposition of their Cephemñnges. Therefore the use of an unnecessarily large Übersehusses at hydrogen peroxide is unwanted. The reaction for the production of the 7-Aminozs of cephalosporansäuredeñvates by conversion of the n (aKetoadipoyl) - 7-ACA-Derivates with hydrogen peroxide runs best at sour pH in the Béíeich from 2,0 to 6.0, vo ugsweise from 3,0 to 5,5. It is very favourable that the optimal pH range of this reaction is essentially a0 same, like that the reaction for the production of the aKetoadipoyl-7-ACA-Deñvates from the Cephalosporinverbindung. It is of advantage further that the MetaUsalz or the organic base, as it is not used gewünschtenfalls with the production of the 7-Aminocephalosporansäuredefivates out as Cephalospoñnverbindung, harmful effect on the reaction for the production of the 7-Aminocephalosporansäurederivates by treatment of the C - Ketoadipoyl-7-ACA-Derivates with hydrogen peroxide exercises. At alkaline pH value the Reakfionsgeschwindigkeit decreases “- Ketoadipoyl-7-ACA-Defivates with hydrogen peroxide and the hydrogen peroxide decompose in the Reaktiousgemisch. On the other hand a-Ketoadipoyl-7-ACA-Deñvat is unstable, with the result at a sanren pH value below 2,0 that the yield at the desired 7-Aminocephalosporansäuredeñvat s in strong extent decreases wird.

The use of the hydrogen peroxide can take place in such a manner that it after the education of the n (A-Ketoadipoyl) - 7-ACADerivates from the Cephalosporinverbindung is added, or it can also equal to beginning of the reaction of the Cephaloso spofinverbindung with “- Ketodeñvat present to be, so that the received OE-Ketoadipoyl-7-ACA-Defivat is transferred into the 7-Aminocephalosporansäuredeñvat. Since a-Ketoadipoyl-7-ACA-Defivat, how it will receive as intermediate product, is chemically unstable, it is not always desired, Wasss serstoffperoxid with this Deñvat after the cleaning for reaction too bñngen.

The reaction broken off by Emiedrigung of the pH value of the reaction liquid below the value of 2,0. In case of the use of hydrogen peroxide this reacts with the 7-Aminocephalosporansäurederivat under training of secondary reactions. Therefore it is desired to remove the surplus hydrogen peroxide previous by reduction. Meanwhile the addition of the usual Redukfiousagenzien causes conversions with the 6s as Ausgan product used Cephalosporinverbindung, as well as with the received 7-Aminocephalosporansäurederivat under change of the Cephemfings. It proved that îür this purpose the use of Thioschwefelsäure or their salts favourably ist.

After a particularly favourable Ausfülmmgsform of the invention the Cephal0sp0rinverbindung I treated with a C - a Ketoderivat in presence of hydrogen peroxide and according to it in an isolated manner in presence of a Thiosulfats, whereby the desired 7-_Arninocephalosporansäurederivat] II selectively and with ausserordeutlich high yields to be received kann.

At most preferential Ausfühnmgsform of the ed_indungsgemässen procedure the conversion of an aqueous solution, the Cephalosporinverbindung, is containing with a A-Ketodeñvat in presence of hydrogen peroxide, on which one converts with a Thiosulfat, for production of the 7-Aminocephalosporansäurederivates. In accordance with these Ausführtmgsform can the 7-Aminocephalosporansäurededvat selektir be received by dissolving hydrogen peroxide in an aqueous solution, which contains the Cephalosporinverbindung, to addition of a ct-Ketoderivates to this solution, conversion of the received mixture during a certain time and afterwards addition from Thioschwefelsäure or a salt from it to the Reaktionsflüssigkeit.

Of this execution form it is characteristic that the Cephalospofinverbindung is converted with a ct-Ketoderivat in presence of hydrogen peroxide, on which the conversion with the Thiosulfat takes place. One receives seleklåv the 7-Aminocephalosporausäuredefivat without Nebenprodukte.

With this execution form the portion of the Thiosulfats hangs, like it the Reakfionsgemisch is added, in certain extent off by the remaining Reaktiousbedingungen, e.g. used the quantity hydrogen peroxide in the Reakfionslösung, the pH value, the Reaktionstemperamr as well as by axe and concentration of the Cephalosporinverbindung used as Ausgangsmatefial. Meanwhile it is usually mindestem äquimolar the used quantity of H20 according to invention received the 7-Aminocephalosporansäuredeñvat can in usual procedures be cleaned, e.g. by extraction from the Reaktionslõstmg using an organic solvent, like ethyl acetate, butyl acetate or n-Butanol.

A further Charaktefistikum of the procedure according to invention consists of the fact that the reaction can be implemented in an aqueous solution. Therefore the 7Aminocephalosporausäuredeñvat can be received, directly e.g.

from a cultural liquid of the Cephalosporinverbindung, without their previous isolation and cleaning. From Standpunk' the t of a commercial Durctfführbarkeit this is extreme vorteilhaft.

In such a way received 7-Aminocephalosporansäurederivate can be transferred, e.g. by application of the Iminoäthermethode, in 3-substituierte Methyl-715-Aminocephalosporansäuren, like it itself among other things as Ausgangsmateñalien for the production of Pharrnaceutica eignen.

Example 1 production of the intermediate product IV 3 g of crystals of the Natrinmsalzes of Cephalospofin C with a purity of 74,2%, according to 2.2 g fine substance, solved in 100 valley water. The received solution added under agitating to 200 valley of an aqueous solution, which contained 4.7 g Glyoxylsäure, 0.3 g copper acetate and 41 mi Pyñdin. The mixture was long agitated at room temperature one hour and decreased after the pH value of the solution by addition by hydrochloric acid under ice cooling on 2,0, for completion of the reaction. Now the reaction solution was combined five times with approximately 200 mi cold ethyl acetate extracted and the excerpts. They were restricted under decreased pressure below 40°C on a volume of approximately 200 ml and after over night with 5°C using water-free Natñumsulfat getrocknet.

Subsequently, the solution was filtered and the arrears washed with a small quantity ethyl acetate. Filtrate and washing solutions were combined and the received solution under decreased pressure below -40°C on a volume was rapidly restricted of 20 valley. Subsequently, it to introduce in drops calmly in 120 mi petroleum ether and under strong Rühren.

The liquid was cooled and centrifuged, on which the supernatant solution was abdekantiert. Whereupon the F'ållung was centrifuged again suspended in petroleum ethers and the received suspension again under cooling. After pouring off the supernatant solution the FäUung over alumina in the vacuum was dried rapidly and resulted in 2.5 g of lightyellow crystals of 3-Acetoxymethyl-71] - (5-carboxy-5-oxopentanamido) - 3-eephem-4-earbonsäure (purity 52%).

TLC: R-S = 0,30; PPC: R-S = 0,52.

621,124 completion of the addition the pH value of the solution 3,5.<br amounted to/> After further 30 minutes one had long agitated, the pH value of the solution was stopped by addition by sulfuric acid atti 1.5, in order to break the reaction off. The solution s was analyzed and contained of 3-Acetoxymethyl-7 [3 (4-carboxybutanamido) - 3-cephem-4-carbonsäure in a portion of 87,8%.

The reaction liquid was treated similar to example 2 and it resulted from it 0.86 g of lightyellow KristaUe of 3lo Acetoxymethyl-7 [3 - (4-carboxybutanamido) -3-oephem-4-carbonsäure. Purity 95,1%.

LS example 2 4 g of crystals of the sodium salt of Cephalosporin C with a purity of 74,2% and 3,3 g copper acetate dissolved in 120 ml water. The pH value of this solution amounted to 3,9. This mixture drop by drop in the process by one hour under agitating at room temperature a 2s aqueous solution was added, which contained 6.24 g Glyoxylsäure and 5.5 ml Pyridin. After completion of the addition the pH value of the solution amounted to 4,6. The pH value was stopped by addition by 1-n sulfuric acid to 3,0, according to which valley 15% ige wässñge hydrogen peroxide solution in the process by one hour into the liquid were introduced in drops. The reaction mixture was long agitated further 10 minutes and adjusted after by addition by sulfuric acid for completion of the reaction to be pH value to 1,5. In this stage the reaction mixture was analyzed and resulted in a raw yield at 3-Acetoxymethyl-7 [3 (4-carboxybutanamido) - 3-cephem-4-carbonsäure of 92%. 3-Acetoxymethyl7 [5 (5-carboxy-5-oxopentanamido) - 3-cephem-4-carbonsäure could not therein discovered werden.

The reaction solution was extracted three times with 500 mi ethyl acetate and the excerpts were combined. They wttrden under decreased pressure and below 40° (3 dried on a volume restricted of 200 ml and after over night with sodium sulfate. The liquid was filtered afterwards and the arrears washed with a small quantity of Äthylace4s did. Filtrate and thereby united washing solution under decreased pressure at a temperature of under 40°C ani a volume of 10 valley were restricted. This concentrate to introduce in drops calmly in 200 mi petroleum ether under strong agitating. The received mixture was centrifuged and the so supernatant solution was abdekantiert. Hernach was again centrifuged the failed product suspended in petroleum ethers and the received suspension. After separating the supernatant solution the F'àllung was dried rapidly over alumina in the vacuum and resulted in 2.38 g lightyellow crystals ss from 3-Acetoxymethyl-7 [3 - (4 - carboxybutanamido) - 3 - cephem4-carbonsäure. Purity 96,9%.

Example 4 from a fermentation solution of Cephalosporitun acremonium the cells were filtered off, on which the solution was stopped to pH 2.8 for decomposition of penicillin N contained in it. following became the solution filtered and the arrears washed. Filtrate and washing solutions were combined and resulted in 4.6 litres of a liquid, which contained 3223 tg/ml Cephalosporin C. To receive this solution given up on a activated charcoal column, which was washed to 700 mi 0,3-n sodium hydroxide afterwards with water and eluiert sehliesslich with 7 litres 3% igem butanol, containing, in order the desired parliamentary group. This parliamentary group was stopped to pH 6.0 and restricted after under decreased pressure at a temperature under 40°C. In this way 275 mi of a lightyellow solution was received, which contained 40 mg/mi Cephalosporin C. In 100 mi of the solution the pH value atti 6.5 adjusted by addition of Natriurnhydroxid, according to which 2.1 g zinc acetate were registered. Then into the solution 500 ml a wässñgen solution introduced in drops by 4,6 g Glyoxylsäure and 9.6 ml y-Picolin in the process of 11/2 hours under agitating at room temperature. During the reaction sodium hydroxide was added, so that the pH value of the liquid amounted to 5.0. In this stage a part of the reaction solution was taken off and subjected to the paper chromatography, whereby it showed up that it 3-Acetoxymethyl-7 - (5-carboxy-5-oxopentanamido) - 3-cephem-4-carbonsäure and 3-Acetoxymethyl-7 [3 - (4-carboxybutanamido) - 3-cephem-4-carbonsäure in raw yields of 61% and/or 18% enthielt.

Afterwards the reaction liquid was long agitated further 30 minutes and adjusted ansehliessend the pH value in it atti 3.0 by addition by sulfuric acid. Now mi 35% iges wässñges Wasserstofîperoxid into the solution were introduced in drops. After further 15 minutes one had long agitated, adjusted in the solution a pH value of 1,5 by addition of sulfuric acid for completion of the reaction. The analysis resulted in a raw to booty of 76,1% at 3-Acetoxymethyl-7 - (4-carboxybutanamido) - 3-cephem-4carbonsäure. 3-Acetoxymethyl-7 [3 (did not know 5 - carboxy-5-oxopentanamido) - 3-cephem-4-carbonsäure in the solution found werden.

The reaction liquid was in the same way treated as in example 2 and resulted in 3.38 g of lightyellow crystals of 3Acetoxyrnethyl-7 [3 (4-carboxybutanamido) - 3-cephem-4-carbonsäure. Purity 78,8%.

Example 3 example 1.5 g of crystals of the sodium salt of Cephalospoñn C with 6o a fermentation solution of Cephalosporium acremonium a purity of 74,2% and 406 mg copper sulfate dissolved by the cells by filtering in 100 mi water, contained in it. The pH value of the solution amounted to liberating and afterwards the pH value adjusted in it on 2,8 3,9. To this liquid 100 valley of an aqueous for decomposition of penicillin N. following was filtñert solution with 2,3 g Glyoxylsäurc and 1.5 ml tri ethyl amine in and the arrears were washed. Filtrate and wash RST portions added by ever 20 ml every 30 minutes. 10 Minu6s sung were combined and resulted in 3 litres of a solution, ten after everyone such addition 3 valley each 20% igen containing 4762 tg/ml Cephalosporin C. in 200 mi of this aqueous hydrogen peroxide solution in addition-given and the solution the pH value atti 7.0 adjusted by addition liquid under agitating 30 minutes react were let. After of sodium hydroxide, according to which 350 mg cobalt sulfate and 621,124,194 mg Piperazin were solved in it and the solution was warmed up to 37°C. Now into the liquid a quantity of 3,4 g Phenylglyoxal in the process was introduced in drops by one hour under agitating. Subsequently, by NatñumhydroxidSchwefelsäure the pH value au/5,0 was stopped. The solution was continued to agitate with 37°C during 30 minutes and adjusted on that by means of sulfuric acid the pH to 3,0. Into this Flüisigkeit 43.9 was introduced in drops ml a satisfied wässdgen sodium perborate solution. After further 10 minutes one had long agitated, the liquid was adjusted by addition by sulfuric acid for completion of the reaction on a pH value of 1,5. Analysis resulted in a raw yield at 3-Acetoxymethyl-7 - (4-carboxybutanamido) - 3-cephem-4carbonsäure of 70,1%.

This Reaktíonslösung aufgearbeítet in accordance with example 2 and resulted in 790 mg of lightyellow Kñstalle of 3-Acetoxymethyl715 (4-carboxybutanamido) - 3-cephem-4-carbonsäure. Purity 72,2%.

Example ó 1.25 g of crystals of the sodium salt of Cephalospoñn C with a purity of 74,2% and 791 mg nickel sulfate dissolved in 100 valley water. To this liquid 2.16 g Glyoxylsäure and 204 mg Imidazol under agitating were added with 5°C. By means of addition of wässñger sodium hydroxide solution the pH value was stopped in the solution to 5,0. After the solution had been long agitated two hours, 53.0 mi of a satisfied wässdgen Kalinmperboratlösung was introduced in drops. Afterwards the solution was long agitated further minutes and adjusted anschliesseud the pH value to 1,5, míttels addition of hydrochloric acid for completion of the reaction. Analysis resulted in a raw yield of 81,4% at 3-Acetoxymethyl-715 (4-carboxybutanamido) - 3-cephem-4carbonsäure.

The reaction mixture regenerated in accordance with example 2 and resulted in 822 mg of lightyellow crystals of 3-Acetoxymethyl715 (4-carboxybutanamido) - 3-cephem-4-carbonsäure. Purity 80,3%.

In exactly same way as managing descriptive, the reaction was accomplished, whereby however in place of nickel sulfate the metal salts indicated in the following table were added. In the table are the salt quantities and the Rohausbetrten received thereby angegeben.

Metal salt rawexploit kind mixes (mg) (mg) FeC13 • 2H20 436,838 A1Cb6H20 725,838 CACI2 • 2I-t20 442,811 FeSO47H20 836,834 MUSO4 • H20 509,831 example 7,2.3 g of crystals of the Nåtriumsalzes of Cephalosporin C with a purity of 74,2% dissolved in 300 mi of a 2m of phosphate buffer of the pH value 4,6. Into this liquid 300 mi of the same buffer solution was registered, which contained 4.7 g Natñumglyoxylat. The addition took place drop by drop in the process from 30 minutes under agitating with 5°C. The solution was agitated two hours long and reacts left, whereby kontinttierlich altogether 33 valley of a 15%igen aqueous hydrogen peroxide solution registered wurden.

Subsequently, further 15 minutes and adjusted then the pH value was long agitated by addition by sulfuric acid for abort of the reaction to 1,5. In this stage analysis of the solution resulted in a raw yield of 76,4% at 3Acetoxymethyl-7 [3 (4-carboxybutanamido) - 3-cepehem-4-carbonsäure.

The reaction mixture was antgearbeitet in accordance with example 2 and resulted in 1.65 g of lightyellow Kñstalle of 3-Acetoxymes thyl-7 - (4-carboxybutanamido) - 3-cephem-4-carbonsäure.

Purity 68.7%.

Example 8,375 mg of crystals of the sodium salt of Cephalospoñn C l0 with a purity of 74,2% and 237 mg nickel sulfate dissolved in 45 mi water. To this solution 620 mg Brenztraubensäure and 61.2 mg Imidazol under agitating were added at room temperature. The pH value was stopped by suitable Ztlgabe of an aqueous Natriumhydroxidlöstmg is August 5.0. After the Flißsigkeit had been long agitated two hours, were drop by drop registered to 15.9 valley of a satisfied aqueous potassium perborate solution. The F! üssigkeit further 301Wmuten was long agitated and after their pH value atti 1.5 by addition adjusted by sulfuric acid for abort of the reaction. In this stage the raw yield at 3-Acetoxymethyl-71 amounted to] - (4-carboxybutanamido) - 3-cephem-4-carbousäure 77,6%.

The reaction mixture wm: regenerated and 238 mg lightyellow crystals at 3-Acetoxymethyl2s 7 - (4-carboxybutanamido) - 3-cephem-4-carbonsäure resulted in de in accordance with example 2. Purity 79,6%.

Example 9 example 2 was done over again, with the difference that ao 3 g of crystals of sodium salt was used by Deaeetyl Cephalosporin C with a purity of 56,3% in place of the sodium salt of Cephalosporin C. From the reaction solution 3 became - In-Iydroxymethyl-7f! - (4-carboxybut anamido) ~3 - cephem-4-carbonsäure in a raw arm booty of 90% enthalas ten. Regenerate resulted in 1.2 g of lightyellow crystals of 3-I-Iydroxymethyl-7 [3 - (4-carbox-ybutanamido) -3 - cephem-4-carbonsäure with a purity of 92,5%.

Example example 2 was done over again, with the difference that g of crystals of sodium salt were used by Deacetoxy Cephalosporin C with a purity of 83% in place of the Natritunsalzes of Cephalosporin C. the reaction ñihrte for 3Methyl-713 (4-carboxybutanamido) - 3-cephem-4-carbonsäure 4s in a raw yield of 95,2%. After regenerating 3.0 g became lightyellow crystals of 3-Methyl-7 [3 (4-carboxybutanamido) - 3-cephem-4-carbonsäure receive with a purity from 96,7%.

thus example ii example 2 was done over again, with the difference that 3 g of crystals of n [7-D (5-Aminoadipinamido) - 3-cephem-3ylmethyl] - pyridinium-4-carbonsäure with a purity were used of 83,9% in place of the sodium salt of ss Cephalospoñn C. the reaction ñihrte for n [7 (4-Carboxybutanamido) -3 - cephem-3 - ylmethyl] - pyñdinium-4earb onsäure in a raw yield of 82,0%. Regenerate the reaction mixture resulted in 2.1 g of lightyellow crystals of n [7 (4-Carboxybutanamido) - 3-cephem-3-ylmethyl] - pyridinium-4-carbonsäure o with a purity of 86,0%.

Example 12 example 7 was done over again, with the difference that 6s 4.6 g Glyoxylsäureamid were used in place of Natriumglyoxylat. The reaction ñihrte to the 3-Acetoxymethyl-715 (4carboxybutanamido) - 3-cephem-4-carbonsäure in a raw yield of 75,9%. With regenerating 1.65 became g of lightyellow crystals of 3-Acetoxymethyl-713 (4-carboxybutanamido) - 3-cephem-4-carbonsäure receive with a purity from 67,9%.

Example 13,1.3 g of crystals of the Natrinmsalzes of Cephalosporin C with a purity of 74,2% dissolved in 250 valley Wasser.

To this solution 441 mg Kupfersulgat were admitted and the pH value was stopped to 5,0. 506 mg Glyoxylsäure under agitating became subsequently, at room temperature zugegeben.

After two hours it had been long agitated the pH value was brought to the solution by means of 1-n sulfuric acid on 3,0, on which 1.9 was registered ml to a 15%igen aqueous hydrogen peroxide solution. Further 10 minutes were long agitated and brought ansehliessend the pH value by addition by sulfuric acid on 1,5, for abort of the reaction. In this stage the solution contained a raw yield of 77,3% at 3-Acetoxymethyl-71 - (4-carboxybutanamido-3-cephem-4-earbonsäure.

Regenerate in accordance with example 2 resulted in 892 mg lightyellow crystals of 3-Acetoxymethyl-7 [3 (4-carboxybutanamido) - 3cephem-4-carbonsäure. Purity 68.8%.

62I 124 earboxybutanamido) - 3-eephem-4-carbonsäure. Purity 68,7%.

Example! 6 s 6.7 g of crystals of the Natñumsalzes of Cephalosporin C with a purity of 51,3% and 1,6 g copper acetate dissolved in 600 valley of a 2-n of phosphate buffer of the pH 4,6. Introduced in drops into this liquid 600 valley of the same phosphate buffer, containing 8.4 g Natñum ct keto n butanoat gradually in lo the process of 30 minutes under agitating with 5°C. The solution was agitated and reacts left, during minutes, whereby gradually 30 wässñgen ml a 35%igen hydrogen peroxide solution was registered. Hernach was long agitated further 10 minutes and afterwards priis the value on 1,5 decreased by addition of sulfuric acid for abort of the reaction. In this stage the analysis resulted in a raw to booty of 63,2% at 3-Acetoxymethyl7 - (4-carboxybutanamido) - 3-cephem-4-carbonsäure in the Reaktionsgemisch.

Regenerate the mixture in accordance with example 2 resulted in 1.92 g of lightyellow crystals of 3-Acetoxymethyl-7 [3 (4-earboxybutanamido) - 3-cephem-4-carbonsäure. Purity 66.3%.

Example 14,1.9 g of crystals of the Natñumsalzes of Cephalosporin C with zs a purity 74.2% dissolved in 350 mi water. To this solution 2 valley Pyridin was admitted and the pH value was brought in it on 4,6. Now 736 mg Glyoxylsäure under agitating were admitted at room temperature. After two hours it had been long agitated pHso the value was brought to the solution by means of 1-n sulfuric acid on 3,0, on which 2.5 mi igen 25% wässñgen hydrogen peroxide solution was registered. The solution was agitated further 10 minutes long and adjusted after the pH value in it by means of sulfuric acid to 1,5, for abort of the Reakas tion. In this stage the Reaktionsftüssigkeit contained a raw yield of 77,1% at 3-Acetoxymethyl-7 [3 (4-carboxybutanamido) - 3-cephem-4-earbonsäure.

Regenerate in accordance with example 2 resulted in 1.3 g of lightyellow crystals of 3-Acetoxymethyl-7 [3 (4-caboxybutanamido) - 3cephem-4-carbonsäure. Purity 68.3%.

Example a fermentation solution of Cephalosporinm acremonium was released from the cells contained in it verrnittels filtration, brought afterwards the pH value in it with acid on 2,8, filtered for decomposition of the Perncillins N contained in the solution. Ansehliessend and washed the arrears. The filtrate was combined with the washing solutions so and resulted in 3 litres of a solution, which contained 4762 tg/ml Cephalosporin C. In 100 valley of this solution was brought the pH value by addition by Natriunflaydroxid on 7,0. Then 175 mg cobalt sulfate and 97 mg Piperazin were solved in it and the solution to 37° (2 warmed up. Now wutss the 1.9 g ct-Ketophenylessigsäure registered in the process from one hour under Riihren and maintenance of a pH value of 5,0 into the solution. Further 30 minutes were agitated with 37oC and adjusted then the pH value to 3,0. Afterwards 22 mi of a satisfied aqueous Natriumperbo “o was introduced in drops raflösung. After further 10 minutes one had long agitated, the pH value of the solution atti 1.5 brought by addition of sulfuric acid for abort of the reaction. In this stage the reaction liquid contained a raw yield of 60,3% at 3-Aeetoxyrnethyl-713 (4carboxybutanamido) - 3-cephem-4-carbonsäure.

Regenerate the reaction solution in accordance with example 2 resulted in 355 mg of lightyellow crystals of 3-Aeetoxymethyl-7 [3 (4Beispiet 17 4 g of crystals of the Natñumsalzes of Cephalospoñn C with a purity of 74,2% and 3,3 g copper acetate dissolved in 120 mi water. The pH value of the solution amounted to 3,9. 120 mi of an aqueous solution from 6,2 g Glyoxylsäure and 5.5 ml Pyridin was registered drop by drop in the process from one hour under agitating at room temperature in it. After completion of the addition the pH value amounted to 4,6.

Now the pH value adjusted to 3,0 by addition of 1-n sulfuric acid, according to which 60 mi 15% ige aqueous hydrogen peroxide solution in the process was introduced in drops by one hour. Afterwards the solution was long agitated still 10 minutes, agitated transferred with 2t, 9 g Natñumthiosulfat, further minutes long and schiiesslich the pH value to 1,5 with sulfuric acid adjusted for abort of the reaction. In this stage the reaction liquid contained a raw yield of 92% at 3-Acetoxymethyl-7l% (4-carboxybutanamido) - 3-eephem-4-carbonsäure. 3-Aeetoxymethyl-7 [3 (5carboxy-5 - oro pent anarnido) - 3-eephem-4-earbonsäure could not in the solution found werden.

The reaction solution was extracted three times with 500 ml Äthy] acetate and the excerpts were combined. It under decreased pressure and below 40°C concentrated on a volume of approximately 200 mi. Ansehliessend over night dried with water-free sodium sulfate. The solution was filtered and the arrears washed with a small portion ethyl acetate. The filtrate and the Waschiösung were combined, according to which under decreased pressure below 40°C on a volume of 10 valley was restricted. This solution introduced in drops in 200 valley petroleum ether under strong agitating. The F'åUung was abzentñfugiert and the supernatant solution was abdekantiert. Subsequently, the precipitation was suspended again in petroleum ethers and the received suspension was again centrifuged. After Abdekantieren of the supernatant solution the precipitation rapidly dried over alumina in the vacuum and resulted in 2.4 g of lightyellow crystals of 3-Acetoxymethyl-7 - (4-earboxybutanamido) - 3-cephem4-earbonsäure. Purity 98,9%.

Example 18,1.5 g of crystals of the Natrinmsalzes of Cephalosporin C with 6s of one pure-preserve from 74,2% and 406 mg copper sulfate dissolved in 100 valley water. The pH value of the solution amounted to 3,9. This liquid 100 mi of an aqueous solution became from 2,3 g Glyoxylsäure and 1.5 valley tri ethyl amine under 621,124 agitating at room temperature in portions of 20 each valley all minutes hinzugegebeu. 10 minutes after everyone this addition were added 3 ml to 20%iges of wässñges Wasserstoffperoxìd and the liquid under agitating was let 30 minutes long react. After completion of the addition the pH value amounted to 3,5. After further 30 minutes it had been long agitated the liquid transferred with 6,29 g Natriumthiosulfat, on which further 10 minutes long agitated wurde.

Subsequently, the pH value adjusted to 1,5 verraittels sulfuric acid for abort of the reaction. In this stage the reaction solution contained a raw to booty of 87,8 g at 3-Acetoxymethyl-7 [5 (4-carboxybutanamido) - 3cephem-4-carbonsäure.

Regenerate in accordance with example 17 resulted in 0.88 g of lightyellow crystals of 3-Acetoxymethyl-7 [5 (4-carboxybutanamido) - 3cephem-4-carbonsäure. Purity 97,1%.

Example 19 4 g of crystals of the sodium salt of Cephalosporin C with a purity of 74,2% and 3,3 g copper acetate dissolved in 120 valley water. The pH value of the solution amounted to 3,9. Into this solution 120 valley of an aqueous solution from 6,24 g Glyoxylsäure and 5.5 valley Pyridin was registered drop by drop in the process from one hour under agitating at room temperature. After completion of the addition the pH value amounted to 4,6. It was adjusted by means of 1-n sulfuric acid to 3,0, on which 60 mi igen 15% wässñgen hydrogen peroxide solution in the process introduced in drops by one hour wurden.

The reaction mixture was long agitated further 10 minutes, according to which 21.9 g Natñurntlúosulfat pentahydrat in it were registered. Ansehliessend was long agitated further 10 minutes and after the pH value brought with sulfuric acid aoE 1.5 for abort of the reaction. In this stage the Reaktionsmischtmg contained a raw to booty of 92% at 3-Aeetoxymethyl-7 - (4-carboxybutanamido) - 3cephem-4-carbonsänre. 3-Acetoxymethyl-7 - (5-carboxy-5oxopentanamido) - 3-cephem-4-carbonsäure could not therein found werden.

The reaction mixture three times with ethyl acetate and the excerpts became extracted - combines. They were restricted under decreased pressure and below 40°C on a volume of approximately 200 valley and after over night dried over water-free Natñumsulfat. Subsequently, and the arrears were filtñert were washed with a small portion ethyl acetate. Filtrate and arrears were combined and restricted under decreased pressure at a temperature under 40°C on a volume of 10 mi. This solution introduced in drops in 200 mi petroleum ether under strong Rühren.

Ansehliessend was abzentrifugiert the precipitation and the überz0 standing solution was abdekantiert. Hernach was again centrifuged the precipitation again suspended in petroleum ethers and the received suspension. After Abtrermung of the supernatant solution the FäUung rapidly dried over alumina in the vacuum and resulted in 2.43 g of hay-yellow crystals of 3-Acetoxyis methyl-713 - (4-carboxybutanamido) - 3-cephem-4-carbonsäure.

Purity 98.9%.

Example 1.5 g KñstaUe of the Natñumsalzes of Cephalospoñn C with a purity of 74,2% and 406 mg Kupferstflfat dissolved in 100 mi water. The pH value of this solution amounted to 3,9. It 100 mi of a wässñgen solution became from 2,3 g Glyoxylsäure and 1.5 mi Tñäthylamln in portions: on ever 20 mi every 30 minutes under agitating at room temperature zs in addition-set. 10 minutes added after everyone this addition ever 3 valley 20% igen Wasserstoffperoxidlöstmg and the liquid during 30 minutes under agitating react let. After completion of the addition the pH value amounted to 3,5. After further 30 minutes one had long geñihrt, the reaction mixture transferred with 6,29 g Natriumthiosulfat pentahydrat. Subsequently, further 10 minutes long one agitated. Hernach the pH value brought with sulfuric acid on 1,5 for abort of the reaction. In this stage the reaction liquid contained a raw yield 3s of 87,8% at 3-Acetoxymethyl-713 (4-Carboxybutanamido) - 3-eephem-4carbonsäure.

The Reaktiousgemisch regenerated in accordance with example 19 and resulted in 0.88 g of lightyellow crystals of 3-Acetoxymethyl713 - (4-carboxybutanamido) - 3-cephem-4-carbonsäure. Purity 97.1%.