PROCESS FOR PRODUCING 3-CEPHEM-4-CARBOXYLIC ACID DERIVATIVES SUBSTITUTED IN 3-POSITION

Abstract: The subject matter of the invention is a process for producing 3-cephem-4-carboxylic acid derivatives sub­ stituted in 3-position of formula (I) wherein R₂ is thienyl, diazolyl, thiazolyl, tetrazolyl, thia­ diazolyl, thiatriazolyl, oxazolyl, oxadiazolyl, pyridyl, pyrimi­ dinyl, benzothiazolyl, benzimidazolyl, benzoxazolyl or any of their substituted derivatives in any of the possible positions of the mentioned heterocyclic residues available for substitu­ tion, which is characterized in that a 7-amino-­ cephalosporanic acid derivative of formula (II) wherein R₁ means -OOCCH₂ is reacted with a thiol R₂SH. The compounds I may be effectively purified by adsorption on a strongly basic anion exchange resin. The compounds I which are obtained by the process of the invention are valuable intermediates in the synthesis of therapeutically useful cephalosporins.

Description: A process for producing 3-cephem-4-carboxylic acid derivatives substituted in 3-position. This invention relates to a valuable method for the synthesis and purification of compounds with the "cephem" structure substituted in the 3 position, which are used as starting products in the synthesis of substances so much appreciated in the pharmaceutical field as the so-called cephalosporins of "second" and "third" generation. EMI1.1 Basically the process according to the present invention is carried out according to the following scheme, in which a molecule of acetic acid is liberated. EMI1.2 where R1 means the -OOCCH3 group and R2 is thienyl, diazolyl, triazolyl, tetrazolyl, thiazolyl, thiadiazolyl, thiatriazolyl, oxazolyl, oxadiazolyl, pyridyl, pyrimidinyl, benzothiazolyl, benzimidazolyl, benzoxazolyl or their substituted derivatives in any of the possible positions of the mentioned heterocyclic structure, available for substitution. A second important aspect of the process is the fact that a highly selective purification procedure is utilized, which allows to obtain in a single step substances of adequate quality for a subsequent use by the pharmaceutical industry. It is well-known by the experts in the art that the use of impure intermediate products renders the purification of the finally synthetized cephalosporins for reaching an adequate degree of quality compatible with the pharmaceutical requirements extraordinarily difficult. Therefore it is necessary to take care of such complex problems as: - Level of unreacted starting materials - Separation of degradation products and products of side reactions from the synthesis itself, for example the A2 isomer obtained by shift of the double bond existing between the positions 3 and 4 of the "cephem" structure to the position 2-3 (IV), the desace tyl-lactone, formed by internal cyclization of the carboxyl of the position 4 with the substituent of the position 3 (V). EMI2.1 and of other degradation products, not absolutely identified, coloured or not, which are a serious problem in the subsequent production steps. It is the objet of the invention to provide a novel process for producing in an excellent degree of purity 3-cephem-4carboxylic acid derivatives substituted in 3-position which are valuable intermediates in the synthesis of therapeutically useful cephalosporins. This object is solved by the surprising finding that by performing a step of adsorption-elution using a macroreti an cular strongly basic anion exchange resin,/excellent degree of purity can be obtained. The subject matter of the invention is a process for producing a compound of the general formula I EMI3.1 wherin R2 is thienyl, diazolyl, thiazolyl, tetrazolyl, thiadiazolyl, thiatriazolyl, oxazolyl, oxadiazolyl, pyridyl, pyrimidinyl, benzothiazolyl, benzimidazolyl, benzoxazolyl or any of their substituted derivatives in any of the possible positions of the mentioned heterocyclic residues available for substitution,which is characterized in that a) a 7- aminocephalosporanic acid (7-ACA) derivative of the general formula II EMI3.2 wherein R1 means OOCCH3 is reacted in an aqueous medium, at a pH between 5,0 and 8,0 and at a temperature between 600C and 800C with a thiole compound of the general for mula III R - SH (III) wherein R2 is as defined above, and b) the reaction product obtained in the previous step is purified by means of adsorption over a macroreticular, strongly basic anion exchange resin having a crosslinked, acrylic copolymer structure and containing functional quaternary ammonium groups. A particular advantage of the process of the invention is the fact that synthesis and purification are carried out in aqLo'us medium. Thus, the inconvenience of a content of residual solvents in the obtained products., which leads to bad consequences regarding the stability, especially long term stability, and imposes decomposition of the pharmaceutically active compounds, is obviated. According to the process of the invention the 7-aminocephalosporanic acid (7-ACA), also known as 7-amino-3-cephem4-carboxylic acid is reacted with a compound from the group of the R2SH thiols1 where R2 means, as previously mentioned, thienyl, diazolyl, triazolyl, tetrazolyl, thiazolyl, thiadiazolyl, thiatriazolyl, oxazolyl, oxadiazolyl, pyridyl, pyrimidinyl, benzothiazolyl, benzimidazolyl, benzoxazolyl, or with a derivative thereof, especially alkyl substituted derivatives, wherein the substituent may be in any position of the heterocyclic structure which is available. The process is a general one and is carried out in an aqueous medium, dissolving the 7-aminocephalosporanic acid and the correspondent thiol in the vehicle of the reaction by addition of an alkali metal hydroxide, ammonium hydroxide or basic compounds which forms water soluble salts; evidently the 7-aminocephalosporanic acid and the thiols can be transformed in to soluble salts by reacting them with an alkali metal carbonate or bicarbonate, an ammonium salt or in general with any salt which can be used for this reaction, without changing the fundamentals of this process. A second way can be the direct use of the soluble salts of the 7-ACA and thiols if commercially available, and this does not change the essentials of the process. In a general way the process is carried out by preparing an aquous solution of 7-aminocephalosporanic acid and the of the respective thiol, in separate recipients or jointly and heating the mixture to an elevated temperature to achieve the condensation in the position 3 of the "cephem" structure and obtaining the-7-amino-3-(substituted)-3-cephem-4-carboxylic acids (I). The variables of the reaction which should preferrably be controlled are the following: - Reaction concentration: is a variable considered slightly critical and generally concentrations between 3 and 10 per cent are preferred. - A second important variable is the R2SH/7-ACA molar ratio and the values which can be utilized are between 1,0 and 2,0, preferably between 1,1 and 1,4 mol R2SH per mol of 7-ACA. - The conditions of relative acidity in which the reaction ta kes place are considered as a third variable, which is li mited by the stability of the B-lactam ring of the 7-ACA and of the 7-amino-3- (substituted) -3-cephem-4-carboxylic acid product of the reaction and by the need of maintaining the reactants in solution. The adequate pH values are those comprised between 5 and 8 and preferably between 6,5 and 7,5. The pH of the reaction mixture may be controlled within the mentioned limits by previous adjustment, or in some cases in which it is modified during the condensation by continuous addition of alkali or by use of buffer solutions; usually the previous adjustment is enough since in the condensation there is no sensible modification the pH of the reaction mixture which is already buffered by itself. The time and temperature variables present the normal tendencies to accelerate the reaction at a high temperature, and for practical reasons reaction times from 0,5 up to 6 hours and temperatures from 60 up to 800C are preferred. Once finished the reaction period the reaction mixture is cooled and acidified up to pH values between 2 and 3,5 preferably with strong mineral acids, such as hydrogen halides or oxy acids. A crude product is precipitated which can be subjected to the subsequent purification step. Equally aceptable are two other variants: a procedure according to which the product of the previous reaction, at a pH between 6,5 and 7,5 is adsorbed over the ionic exchange resin directly, without the mediation of the crude compound precipitation step, or a procedure according to which the precipitated compound, as mentioned in the above paragraph, is suspended in water to be again dissolved by addition of the quantities of alkali needed to bring the solution to the adequate pH which oscillates from 6,5 up 8,5. It has been found that a macroreticular, strongly basic anion exchange resin having a crosslinked acrylic copolymer structure and containing functional quaternary ammonium groups is preferred in the sense that is produces practically quantitive results and a high degree of purification. The quantity of the resin is based on the consideration that the theoretical exchange capacity of the mentioned resin is in the order of 1-1,2 meq/ml. Depending on the height/diameter ratio of the column and on the degree of the tolerated losses, a quantity of between 3 and 10 ml of resin/gram of compound to be adsorbed will be utilized. The resin is used in the acetate state which may be obtained - following the indications of the manufacturer to convert it to the free base state - by treating it in re-cycle with a quantity of acetic acid in the order of 1,2-1,5 equiv/l of resin, for assuring a total conversion to the acetate state. Right after this the resin is carefully washed and adjusted in batch to the pH required for the posterior adsorption by addition of sodium hydroxyde. Than the solution to be purified is passed over the pretreated resin at such flows that the time of contact is between 20 and 40 minutes, in descending flow. Finished the adsorption, the column is displaced with 2-3 volumes of water. The use of an ionic exchange resin allows the elimination of non-ionic impurities which might be present (lactone type), which are hardly eliminated by any other procedure. Among the different types of eluents the use of a sodium acetate buffer solution with a concentration of 0,1-0,5 M and a pH of 7-7,5 is preferred. From the eluate so obtained, the product of the invention is isolated according to conventional acidification methods. The following examples illustrate the present invention. EXAMPLE No 1 12,76 g of 1-methyl-5-mercapto-1,2,3,4-tetrazol were dissolved in 300 ml of water, the pH being adjusted at 6,6 by means to an ammonium hydroxide solution and the solution was heated at 70"C. Simultaneously 29,6 g of 7-ACA were dissolved in 400 ml of water by means of addition of ammonium hydroxide up to pH 7,5. The mentioned solutions were mixed up and stirred for 70 minutes at 680C, adding the ammonium hydroxyde necessary to keep the pH between 6,8 and 7,0. It was quickly cooled to 25"C and the pH was adjusted at 2,5 with concentrated hydrochlo there ric acid. Immediately/after it was cooled at 0-5"C and crys- tallization was performed for 1 hcur.The compound was filtrated and washed with 125 ml of water and 350 ml of acetone. 24,2 g of the 7-amino-3-(1-methyl-5-mercapto-1,2,3,4-tetrazolyl)-3-cephem- 4-carboxylic acid were obtained identified by its IR spectrum, according to the standard. EXAMPLE No 2 2,14 kg of 2-methyl-5-mercapto-1,3,4-thiadiazol were dissolved in 72 1 of water by addition of 1,65 kg of triethyl amine and stirred at room temperature for half an hour. Simultaneously 4,0 kg of 7-ACA were dissolved in 72 1 of water, by addition of 1,51 kg of triethylamine and stirred at room temperature for half an hour. The 2-methyl-5-mercapto-1,3,4-thiadiazol solution was heated and the 7-ACA solution was added with vigorous stirring. It was heated quickly up to a maximum temperature of 72"C and was kept at these conditions for 1 hour.Imme there diately/after it was cooled to 250C, the pH was adjusted to 2,5 by addition of 3,3 kg of concentrated hydrochloric acid and finally the compound was crystallized by stirring at 50C for 1 hour. It was filtrated and washed with 20 1 of water, 60 1 of acetone at 80t and 20 1 of acetone. By this way 4,3 kg of 7-ami no-3- (2-methyl-5-mercapto-1,3,4-thiadiazolyl)-3-cephem-4-carbo- xylic acid were obtained, with a purity of the 85 percent and identified by its IR spectrum according to the standard. EXAMPLE No 3 32 g of 2-methyl-5-mercapto-1,3,4-thiadiazol were dissolved in 350 ml of water, adjusting the pH at 6,8 with ammonium hydroxyde and the solution was heated to 750C. Simultaneously 51 g of 7-ACA were dissolved in 720 ml of water by addition of ammonium hydroxyde up to pH 7,5. The solutions were mixed-up, the temperature of the mixture was adjusted to 780C and the p14 to 6,0-7,0, keeping the mentioned conditions for 90 minutes. It was cooled quickly to 250C and was crystallized by adjusting the pH to 2,5 by addition of hydrochloric acid. The crystallization was prolonged for 1 hour of stirring at 0,5"C. The obtained com pound was filtrated and washed with 250 ml of water and 3 portions of 250 ml of 808 acetone each one. By this way 51,4 g of the 7mino-3(2-methyl-5-mercapto-1,3,4-thiadiazolyl)-3-cephem 4-carboxylic acid were obtained. 50 g of this compound were dissolved in 1 1 of water by addition of ammonium hydroxyde up to pH 7,5. This solution was passed through a column loaded with 500 ml of AMBERLITA IRA-958 resin (manufactured by Rohm and Haas) in the acetate cycle, at an flow of 20 ml/min, discarding the efluent, It was eluted with sodium acetate buffer 0,4 M, pH 7,5 at a flow of 13 ml/min, retaking the central fraction of eluate, considering as such that in which precipitate appears upon aci dification,obtaining 1.460 ml of eluate. From this eluate which contains the 7-amino-3(2-methyl-5-mercapto-1,3,4-thiadia- zolyl)-3-cephem-4-carboxylic acid purified and in the form of sodium salt, the compound was crystallized in acid form by adjusting the pH at 2,5 by addition of concentrated hydrochloric acid. It was filtered, washed, and dried according to the conventional procedures, obtaining 40,8 g of compound with a purity higher than 96 percent. Identification, IR (KBr) cm 1 , 1.803, 1.618; 3,35 (1H, d, JHbHd2 9,3 Hz) 3,74 (1H, d, JHaHc= 12,45 Hz) 3,86 (1H, d, JHdHb= 9,3) 4,44 (1H, d, JHcHa= 12,45 Hz) 4,68 (1H, d, JH6H7= 4,8 Hz) 4,95 (1H, d, JH2H6= 4,8 Hz) EMI10.1 EXAMPLE No 4 12,76 g of 1-methyl-5-mercapto-1,2,3,4,-tetrazol were dissolved in 300 ml of water, being adjusted the pH at 6,6 by means of ammonium bydroxyde and the solution was heated to 70"C. Simultaneously 29,6 g of 7-ACA were dissolved in 400 ml of water by addition of ammonium hydroxyde up to pH 7,5. The mentioned solutions were mixed up and the stirring was kept for 70 minutes at 68"C, adding the ammonium hydroxyde necessary to keep the pH between 6,8 and 7,0. It was quickly cooled to 250C and the pH was adjusted at 2,5 with concentrated hydrochloric acid. Immedia there tely/after it was cooled to 0-50C and the crystallization was performed for 1 hcur with stiring.The compound was filtered and was7 hed with 12 ml of water and 350 ml of acetone. 24,2 g of 7-ami no-3-(1-methyl-5-mercapto-1,2,3,4-tetrazolyl)-3-cephem-4-carbo- xylic acid were obtained. This crude compound was dissolved in 0,5 1 of water by addition of ammonium hydroxyde up to steady pH 7,5. This solution was passed through a column loaded with 250 ml of AMBERLITA IRA-958 (manufactured by Rohm and Haas) in the acetate cycle, at a flow of 20ml/min., discarding the efluent The column was displaced with 400 ml of water, at the same flow, discarding the efluent likewise. Then was eluted with sodium acetate buffer 0,4 M, pH 7,5, at a flow of 13 ml/min., taking up the central fraction of the eluate, considering as such that in which a precipitate is formed upon acidifying. 730 ml of eluate were retaken, containing the 7-amino-3- (1-methyl-5mercapto-1,2,3,4-tetrazolyl)-3-cephem-4-carboxylic acid in the form of sodium salt. The free acid was crystallized by adjusting the pH to 2,5 by addition of concentrated hydrochloric acid. It was filtered, washed and dried according to the conventional procedures, being obtained 20,4 g of compound with a purity higher than 96 percent. Identification, Ir (KBr) cm 1 , 1.790, 1.610. NMR (D20+CF3COOD) ppm: 3,82 (2H, s, C2H2) 4,07 (3H, s, N-CH3) 4,41 (1H, d, JHaHb 13,7 Hz) 4,26 (1H, d, JHbHa= 13,7 Hz) 5,26 (1H, d, JH7= 4,8 Hz) 5,19 (1H, d, JH6- 4,8 Hz) EMI12.1

Claims: Claims 1. A process for producing a compound of the general formula I EMI13.1 wherin R2 is thienyl, diazolyl, thiazolyl, tetrazolyl, thiadiazolyl, thiatriazolyl, oxazolyl, oxadiazolyl, pyridyl, pyrimidinyl, benzothiazolyl, benzimidazolyl, benzoxazolyl or any of their substituted derivatives in any of the possible positions of the mentioned heterocyclic residues available for substitution, characterized in that a) a 7- aminocephalosporanic acid (7-ACA) derivative of the general formula II EMI13.2 wherein R1 means -OOCCH3 is reacted in an aqueous medium, at a pH between 5,0 and 8,0 and at a temperature between 60"C and 80"C with a thiole compound of the general for- mula III R2 - SH (III) wherein R2 is as defined above, and b) the reaction product obtained in the previous step is purified by means of adsorption over a macroreticular, strongly basic anion exchange resin having a crosslinked, acrylic copolymer structure and containing functional quaternary ammonium groups. 2. A process according to claim 1characterized in that,the rea ts are added to the reaction mixture as such or in the form of sodium salts, or in the form of any other salt of inorganic or organic base soluble in water. 3. A process according to claim 1 or 2, characterized in that the reactants are employed in a molar ratio of from stoechiometric ratio up to a considerable excess of the R2SH reactant, preferably in a ratio within the range of 1,1 to 1,4 mol of R2SH per mol of 7-ACA. 4. A process according to claims 1 to 3, characterized in that the reaction time is comprised between 0,5 and 6 hours, preferably between 1 and 2 hours. 5. A process according to claims 1 to 4, characterized in that the reaction is carried out at a pH-value between 5 and 8, preferably between 6,5 and 7,5. 6. A process according to claim 1, characterized in that ion exchange resin is in the acetate state. 7. A process according to claims 1 and 6, characterized in that the product is eluted from the resin with a sodium acetate buffer solution having a concentration between 0,1 and 0,5 M and a pH between 7 and 7,5. 8. Use of the compounds of general formula I as obtained according to anyone of claims 1 to 7 for the production of cephalosporin antibiotics.

Inventors: ALBA, D. ALEJANDRO VITALLER, DR., LIZARBE, D. JOSE RAMON FERNANDEZ, DR.
Applicants: ANTIBIOTICOS, S.A.
Application year: 1984
Main classification: C07D501
IPC codes: C12P35, C07D501