FI60210B - FREQUENCY REFRIGERATION FOR 7BETA-ACYLAMIDO-CEFALOSPORANSYRAESTERFOERENINGAR - Google Patents

Description

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Patent application V ^ (51) Kv.lk.3 / lnt.CI.3 C 07 D 501/04 SUOMI-FINLAND W) Patent application - Patentansökninf 3301 / "Jk (22) Hakemlspllvi - AntBknlngsdag lU.11.71 * ^ ^ (23 ) Alkuptlvi — Glklghetsdag li ;. 11.Tl (41) Tullut | ulklsaksl - Bllvlt offentllg 31.05.75

Patent and Registration Office ......... .. . ,. ,,, _ ^ ^ ^ ___, (44) Date of publication and month of publication. -

Patent · eeh registerstyrelsen '' AnsMcan utlagd och utl.skrlftan publtcerad 31.08.8l (32) (33) (31) Privilege claimed — Begird priority 30.11.73 23.09.7U USA (US) U20U18, 507U73 (71) Merck & Co. , Inc., Rahway, New Jersey 07065, USA (72) Leonard Maurice Weinstock, Belle Meade, New Jersey, USA (7U) Oy Kolster Ab (5U) Process for the preparation of 7β-acylamido-cephalosporanic acid ester compounds - Förfarande for framställning av 7/3 -acy lami do -cef aio -sporansyraesterföreningar

The present invention relates to an improved process for the preparation of Ifi-acylamido-cephalosporanic acid ester compounds of the formula

R. H

w 1 I 1 rt, le r, nt “T Ί (i> J-" γ1- CH2ä 'COOR "where R1 is hydrogen or methoxy, A * is carbamoyloxy or acetoxy, R' is an acyl group of formula

O

R -CH-C-

3 I

B

6021 0 wherein R3 is phenyl, 2-thienyl or 2-furyl, and R "is an easily removable ester protecting group such as methoxymethyl or benzhydryl. The process of the invention is characterized in that the compound of formula

R

H 1 S -N CH 2 A 'COOR "wherein B is a protected aminoadipoyl group and R 1, R" and A' are as defined above is reacted in a neutral solvent with an acylating agent of formula

O

R .ch-CC1

^ I

B

wherein is the same as above, in the presence of a molecular sieve catalyst of synthetic aluminosilicate zeolite species 3A, 4A, 5A or 13X, having a pore size of about 3-15 A and anhydrous or containing not more than 30% by weight of hydrated water, and after the acylation reaction, the group B is cleaved to give the product of formula (I) either by extending the reaction time or by adding a cleavage agent such as benzyl alcohol, alkanol, lower alkylthiol or hydrochloric acid, or when the molecular sieve contains water of hydration.

Process for the preparation of 7/3-acylamido-cephalosporanic acid ester compounds by acylation of 7/3- (D-5-amino-5-carboxyvaleramido) cephalosporanic acid ester compounds followed by cleavage of the aminoadipoyl group from the diacylated product is described in the present invention. compounds containing a trimethylsilyl group are used as a catalyst in the acylation step. The advantage of the zeolite catalyst system used in the process according to the invention over the silyl catalyst system is that the zeolite system is heterogeneous and 3 60210 gives a larger surface area where the reaction can take place, controlling acid formation (hydrochloric acid is released during transacylation). . The table on page 17 shows the effect of the zeolite catalyst system compared to Example 14 in the table in which no zeolite catalyst is present.

Molecular sieves useful in the context of the invention include aluminosilicate zeolites. Generally speaking, naturally occurring zeolites can be defined as a group of crystalline solids formed by hydrated aluminosilicates of monovalent and divalent bases which are capable of donating part or all of the water they contain without altering their crystal structure, absorbing other compounds instead of effluent. On the other hand, the synthetic zeolite is synthesized from a combination of base oxides (AlO 2, SiO 2 'Na 2 O' K 2 O, 3ne) in an aqueous system to give a hydrated or hemihydrated crystal structure. After heat treatment, the zeolites can be considered to be essentially anhydrous. Synthetic zeolites are characterized and classified mainly by X-ray powder diffraction methods. Although there is no systematic chemical method suitable for naming synthetic aluminosilicate complexes, each new synthetic zeolite is named by letter or group of letters and number. The meaning of these selected symbols is well understood by those skilled in the art.

It has been found that class A and X synthetic zeolites are preferred in the acylation process of the invention. The pore sizes of the zeolite pathways may range from about 3 to 15 A. The zeolites may be substantially anhydrous or may contain some hydrate water. The amount of water contained in the alloy may be 0-30% by weight.

Screen types 3A, 4A, 5A and 13X used in the context of the invention have the following characteristics.

6021 0

Type Formula Pore diameter

3A KqNa12_q <f (AlO 2) 12 (SiO 2)>). 27H 2 O 3A

4A Na2 ^ (A102) 12 (SiO2) ^ .27H2O 4A

5A Ca45Na3 ^ (AlO2) 12 / .30H2O 5A

13X Na86 ^ (A102) g6 (SiO2) y. 106-xH2O 10A

The Type 3A resin is prepared from a Type 4A resin by replacing some of the sodium ions with potassium ions. The term "q" denotes the number 1-12 and indicates how many sodium ions have been replaced by potassium ions. The term "x" in type 13X denotes the number 0-106 and indicates the amount of hydrated water.

The screens are available essentially anhydrous, can be used in this form, or can be further dewatered to 0-12%, by heating at high temperatures (about 500 ° C or higher) before use; or they may be used when they contain about 30% (w / w) hydrate-water. Hydrated screens are prepared by allowing the screens to be in chambers or environments with a high moisture content, or by slurrying in water and then adjusting the moisture content as desired by drying in vacuo or drying at or above room temperature. This drying usually takes about 2-5 hours, although this is not a critical time interval. The moisture content can be determined using the Karl Fischer method or another available method.

The implementation of the method according to the invention is described in more detail below.

5,60210

? 2 R1 H

NH 0 „! 'e 1, 1 H b CH (CH2) 3C-N —I-γ COOR "

^ 0 ^ -NY 'CH2A

(ID C00R "

R R1 H

? 2. 1; NH 0 r. r _

CH (CH2) 3C-N —- Y N

COOR "> n n ^ ~ ch2a · COOR" (III)> r

R, H

j '

,B

R N - <^ J? -N w ^ CH A 'ο £ COOR "(i)

In the above flow chart, R 1, R ', R "and A' have the same meaning as above and R 2 is a protecting group.

6 60210

The amino group of the aminoadipoyl group may be protected with conventional amino protecting groups (R 2, such as acyl, aroyl, alkoxycarbonyl, alkylsulfonyl, arylsulfonyl and other groups known in the art. Suitable groups include, e.g. 2-nitrophenylsulphenyl, 2,4-dinitrophenylsulphenyl, chloroacetyl, p-nitrophenylthio, p-nitrobenzenesulphonyl, p-toluenesulphonyl, methanesulphonyl, benzoyl, p-chlorobenzoyl, p-nitrobenzoyl, benzonyl, sulphonyl is easily prepared by reacting a cephalosporin compound with p-toluenesulfonyl chloride or benzoyl chloride by maintaining the pH of the mixture in a basic range, i.e. between 9 and 10.

In general, it is preferred that the acylation be performed using a cephalosporin compound in which the carboxy groups of both the aminoadipoyl side chain and the 4-position are similarly protected (R "), in which case the product is obtained in maximum yields. thus, the protecting group is essentially one which can be easily removed at the 4-position to give the free acid without cleavage of the β-lactam group, since the cephalosporin compounds are usually used as salts such as an alkali metal salt or an amine salt, and suitable protecting groups are well known to those skilled in the art. suitable derivatives are esters of alcohols, phenols, mercaptans and thiophenols. In the formula (II), R "corresponds to a radical of an alcohol or thiol such as methyl, ethyl, tertiary butyl, substituted alkyl such as phthalimidomethyl, succinimidomethyl, phenacyl, subs substituted phenacyl, for example p-bromophenyl, β-substituted ethyl such as 2,2,2-trichloroethyl, 2-methylthioethyl, 2- (p-methylphenyl) ethyl, 2- (p-methylphenyl) sulfonylethyl, 2- methylaminoethyl, 2-chloro (or bromo) ethyl, benzyl, a substituted benzyl group such as p-nitrobenzyl, p-methoxybenzyl, 3,5-dinitrobenzyl, 2,4,6-trimethylbenzyl, 3,5-dichloro-4-hydroxybenzyl, benzhydryl or a substituted benzhydryl group such as p-methoxybenzhydryl, an acyloxyalkyl group such as acetoxymethyl, pivaloyloxymethyl, 7,60210 alkoxy groups such as methoxymethyl, or a monocyclic aryl group, for example phenyl or substituted phenyl such as phenyl or substituted phenyl such as phenyl or substituted phenyl It has been found that the preferred group used for protecting the carboxy group is a methoxymethyl or benzhydryl group.

The protected cephalosporin compound (II) is reacted with an acylating agent in the presence of a molecular sieve to give the diacylated product (III).

The acylating agent is used in an excess of moles relative to the amount of cephalosporin starting material, with the amount of acylating agent being substantially 2-6 times greater than that of cephalosporin.

Conversion of the protected cephalosporin compound (II) to the diacylated product (III) is accomplished by contacting the cephalosporin compound with the acylating agent in a suitable medium in the presence of a molecular sieve. The temperature at which this reaction is carried out is not critical, and satisfactory temperatures are generally about -20 ° C to + 100 ° C. Since the reaction is temperature dependent and occurs more rapidly at higher temperatures, it is recommended that the reaction be carried out at a temperature of about 50 ° C to 90 ° C. Suitable media for the reaction mixture include various solvents free of active hydrogen such as chloroform, acetonitrile, methylene chloride, dioxane, benzene, halobenzene, carbon tetrachloride, 1,2-dichloroethane and diethyl ether. It is important that the slurry is kept in motion during the reaction by stirring or stirring.

The amount of molecular sieve required for the exchange reaction can be varied according to the selected working conditions. In general, it is recommended that the starting material and the molecular sieve be used in approximately the same weight amounts, although it is possible that the starting compound and the molecular sieve are used in weight ratios of 1: 0.5 to 2.

The aminoadipoyl group B can be cleaved by various methods. In some cases, simply "aging" the reaction mixture is sufficient, e.g., with molecular sieves containing 10-30% water, from 30 minutes to 30 hours. Alkanol, lower alkylthiol or benzyl alcohol may be added, followed by a short "aging" interval. The alkanol or lower alkylthiol has 1 to 6 carbon atoms. These are mainly methanol, ethanol, isopropanol or t-butanol. Cleavage can also be performed with hydrochloric acid. Some happens during the acylation reaction

8 6 C 21 O

cleavage of the aminoadipoyl group, since the reaction is an equilibrium reaction in nature, depending on the conditions under which the acylation is carried out. Prolonged heating of the reaction mixture results in cleavage of the aminoadipoyl group and formation of the desired 7-acylated cephalosporin compound, especially when the molecular sieve contains more than 10% water. Deprotection of the carboxy group is carried out according to known methods. Thus, for example, the methoxymethyl group is removed using hydrochloric acid at a temperature of 0 to 10 ° C, the trichloroethoxycarbonyl group is removed by reaction with zinc and acetic acid; and the t-butoxycarbonyl and benzhydryl groups are removed by reaction with trifluoroacetic acid.

Example 1 3-Carbamoyloxymethyl-7-methoxy-7β-thienylacetamido-3-cephem-4-carboxylic acid ___

Step A: If 3- (D-5-tosylamino-5-carboxivaleramido) -3-carbamoyloxymethyl-7-methoxy-3-cephem-4-carboxylic acid__ 3- (3- (D-5-amino-5-carboxyvalerido) ) -3-Carbamoyloxymethyl-7-methoxy-3-cephem-4-carboxylic acid monosodium salt (45.0 ml of a 49.5 mg / ml aqueous solution) is mixed with acetone (450 ml) and water (450 ml). The pH of the mixture is adjusted to 9.5 to 9.6 with 50% NaOH solution and tosyl chloride (19 g) dissolved in acetone (100 ml) is added portionwise. The pH is maintained between 9.5 and 9.6 by the occasional addition of a basic solution. After 15-20 minutes, the pH remains the same: sulfonylation continues for a total of one hour. The temperature of the solution is 20-23 ° C throughout the reaction.

The solution is then cooled in an ice bath and the pH is lowered to 7 by the addition of HCl (1: 1, ice cold). The solution is extracted with ethyl acetate. The ethyl acetate layer is washed back with 100 ml of 5% sodium chloride solution. The organic layer is discarded and the aqueous layers and the mixture containing 500 ml of ethyl acetate are readjusted to pH 2.5 and the layers are separated. The aqueous layer is extracted 3 more times with 500 ml of ethyl acetate. The ethyl acetate layer is washed back with 100 ml of saturated NaCl solution. The extracts are dried over Na 2 SO 4 and the solution is concentrated to low volume. (Temperature 30 C). The concentrated solution is then dissolved in 200 ml of isopropanol, heated to 6021 DEG-45 DEG C., and 5.8 ml of acetic acid and 21.6 ml of dicyclohexylamine are added.

This slurry is allowed to cool slowly and aged overnight at room temperature. The product is filtered off, washed with 100 ml of isopropanol and dried overnight at room temperature under high vacuum.

The product, 7β- (D-5-tosylamino-5-carboxivaleramido) -3-carbamoyloxymethyl-7-methoxy-3-cephem-4-carboxylic acid, dicyclohexylamine salt, is obtained in 44.5 g yields; UV: (pH 7.0 buffer) λ max 2620 E% 94.7

Equivalent weight (HClO 2 titr.) 481.5 (theoretical 481.5)

Analysis calculated for C 47 H 18 N 2 O 2 • C, 58.60; H, 7.74; N, 8.72;

Found: C, 58.29; H, 7.29; N, 8.73.

Step B: ββ- (D-5-Tosylamino-5-carboxivaleramido) -3-carbamoyloxymethyl-7-methoxy-3-cephem-4-carboxylic acid dimethoxymethyl ester_

The tosyl salt obtained in Step A, 20 g, is placed in a 3-necked flask. Methylene chloride (200 mL) is added and the slurry is cooled to 0 ° C in an ice bath under nitrogen. Chloromethyl methyl ether (4.1 ml) in 30 ml of methylene chloride is added to the reaction mixture over 90 minutes with good stirring and ice-cooling. After one hour of addition, a solution of collidine (1.58 ml) in 5 ml of methylene chloride is added.

After the additions, the mixture is stirred for another two hours, filtered, and the filter cake is washed with dry methylene chloride. After extraction with aqueous solutions of phosphoric acid, sodium chloride, sodium hydrogencarbonate and sodium chloride, the filter cake is washed back with methylene chloride. The organic layer is dried, filtered, concentrated to low volume and crystallized. The product, 7β- (D-5-tosylamino-5-carboxivaleramido) -3-carbamoyloxymethyl-7-methoxy-3-cephem-4-carboxylic acid dimethoxymethyl ester, is obtained in 9.6 g (yield 83.5%). ). Both ultraviolet and plate chromatographic analysis showed that the product contained only one component.

10 6021 0

Step C; 3-carbamoyloxymethyl-7-methoxy - '^ 3-thienyl-acetamido-3-cephem-4-karboksyylihappo_

To a stirred slurry of 6.9 g of the tosylmethoxymethyl ester obtained in Step B and 7.5 g of a powdered molecular sieve type 4A (600 mesh, hydrated to 17% water in 2% water) in 85 mL of 1,2-dichloroethane was added. 5 ml of distilled 2-thienylacetyl chloride. The stirred slurry was heated at 65 ° C for 16 hours under nitrogen.

The reaction is monitored by plate chromatography. After said time, the majority of the reaction mixture is the desired intermediate. Methanol (0.8 ml) is then added and the slurry is aged for a further 2 hours. In this case, the slurry contains essentially a compound which is the 4-methoxymethyl ester of the desired product.

The ester is hydrolyzed by cooling the above solution to 25 ° C, filtering and washing with cold methanol. The filtrate and washings are combined and cooled to 0 ° C. A 0 ° C solution of 20.8 ml of concentrated HCl and 23.6 ml of methanol is added and the solution is warmed to 15 ° C and stirred at 15 ° C for 2 hours 40 min. In this case, a plate chromatographic analysis is performed. The pH of this mixture is first adjusted by passing ethylene oxide through the mixture as needed, between 2 and 2.5; then the pH is raised to between 5-6 by the addition of solid sodium hydroxide. The mixture is filtered and the dichloroethane layer is separated. The cold aqueous layer contains the sodium salt of the product. This layer is purified using column chromatography and IRA-68 resin on a nitrate cycle with an eluent of 0.02 moles of phosphate buffer pH 7.0. The final product yield is 1.74 g; it gives a single spot on the chromatographic plate. The rotation angle is 192 °. The identity of the product is verified using an NMR assay; the product is the desired 3-carbamoyloxymethyl-7-methoxy-γ-thienylacetamido-3-cephem-4-carboxylic acid, mp 165-167 ° C.

Example 2 3-Carbamoyloxymethyl-7-methoxy-7,3-phenylacetamido-3-cephem-4-carboxylic acid__

Step A; 7β- (6D, 5-Tosylamino-5-carboxivaleryl) phenylacetylamido-3-carbamoyloxymethyl-7-methoxy-3-cephem-4-carboxylic acid dimethoxymethyl ester

Solution of ββ- (D-5-tosylamino-5-carboxylvalido) -3-carbamoyloxymethyl-7-methoxy-3-cephem-4-carboxylic acid dimethoxymethyl ester (9.3 g, 10 minol) , a type 12 A powdered molecular sieve (hydrated to 20% * 2% water (10.8 g) and phenylacetyl chloride (5.3 mL, 40 mmol) in 50 mL of acetonitrile, heated at 40 ° C for 20 h. after which time the mixture is cooled to room temperature and filtered, the filtrate is evaporated to dryness and triturated with hexane, an insoluble residue containing N- (D-5-tosylamino-5-carboxyvaleryl) phenylacetylamino] -3-carbamoyloxymethyl-7-methoxy -cephem-4-carboxylic acid dimethoxymethyl ester, used without further purification in the next step.

Step B: 3-Carbamoyloxymethyl-7-methoxy-7H-phenylacetamido-3-cephem-4-carboxylic acid

The crude product obtained in step A is dissolved in 50 ml of 1,2-dichloroethane. 1.0 ml of ethanol are then added and the solution is stirred for one hour. The methoxymethyl ester is hydrolysed by adding a solution of 20 ml of concentrated HCl in 25 ml of methanol at 0 ° C and stirring for 3 hours at 15 ° C. The product is isolated and purified using the same general procedure as in Example 1. The product gives 3-carbamoyloxymethyl-7-methoxy-7H-phenylacetamido-3-cephem-4-carboxylic acid, m.p. is 159-161 ° C, and the UV and NMR spectra are consistent with the reported structure.

Example 3 3-Carbamoyloxymethyl-7-methoxy-N- (2-furylacetamido) -3-cephem-4-carboxylic acid 7- (D-5-tosylamino-5-carboxivaleramido) -3-carbamoyloxymethyl-7-methoxy- Dimethoxymethyl ester of 3-cephem-4-carboxylic acid is reacted with 2-furylacetyl chloride in the presence of 12 g of a hydrated molecular sieve of type 4A (hydrated to 15% → 2% water) following the procedures just described. The side chain and ester blocking group are also removed following the methods described. The product obtained is 3-carbamoyloxymethyl-7-methoxy-N- (2-furylacetamido) -3-cephem-4-carboxylic acid, m.p. 156-161 ° C, UV (pH 7.0 buffer) λ max 265 nm, ε 7200, IR and NMR spectra consistent with structure.

Example 4 3-Acetoxymethyl-N- (2-thienylacetamido) -3-cephem-4-carboxylic acid Step A: β- (D-5-trichloroethoxycarbonylamino-5-carboxalamido) -3-acetoxymethyl-3-cephem- 4-Carboxylic acid To a solution of - (D-5-amino-5-carboxyvaleramido) -3-acetoxymethyl-3-cephem-4-carboxylic acid (2.5 g, 0.53 mol) in acetone (13 mL) 12 6021 0 3a 10% aqueous dipotassium hydrogen phosphate solution (40 ml) is added dropwise to trichloroethoxycarbonyl chloride (3 * 35 St 0.159 mol). During the addition, the pH of the solution is maintained between 8.5 and 9 * 0 by gradually adding 17% aqueous sodium hydroxide solution. After 30 minutes, the mixture is washed with ethyl acetate and the aqueous layer is acidified by adding concentrated hydrochloric acid until the pH is 2.5. · The precipitated product is extracted into ethyl acetate, the solution is dried over sodium sulfate, filtered and the solvent is removed to give 2.7 S 7P- (1> -5 "trichloroethoxycarbonylamino). 5-karboksivalera-mido) -3-acetoxymethyl-3-cephem-4-carboxylic acid.

Step B: 7- (D-5-Trichloro-ethoxycarbonylamino-5-carboxy-valeramido) -3-acetoxymethyl-3-cephem-4-carvedxylic acid dibenzhydryl ester _________________

To a solution of 73- (D-5-trichloroethoxycarbonylamino-5-carboxyvaleramido) -5-acetoxymethyl-3-cephene-4-carboxylic acid in ethyl acetate (30 mL) was added diphenyldiazomethane (2.0 g) dissolved in ether (25 mL). . The mixture is stirred overnight and the solvent is removed to give 4.0 g of crude product. The crude product is purified by chromatography on silica gel using chloroform as the eluent to give 2.3 S of essentially pure 7- (D-5-trichloroethoxycarbonylamino-5-carboxyvaleramido) -3-acetoxymethyl-3-cephem-4-carboxylic acid dibenzhydryl ether. - CLCl 2) 6 ** 2.0 (methyl, s), 4 * 9 (10-Ho, quartet), 3 * 2 (2-H 2, quartet), 4.95 (6-H, d), 5 , 92 (7-H), 7.0 (benzhydryl protons, 2 s),

Step C: 7 - [(D-5-Trichloro-ethoxycarbonylamino-5-carboxy-valeryl) -2-thienyl-acetylamino] -3-α-ethoxymethyl-3-cephem-4-carboxylic acid dibenzhydryl ester_________

A mixture of 7β- (h-5-trichloroethoxycarbonylamino-5-carboxyvaleramido) -3-acetoxymethyl-3-cephem-4-carboxylic acid dibenzhydryl ether (2.0 g, 0.02 mol), 11.0 g A type 5A molecular sieve (hydrated 23 fo to $ 2 containing water), 2-thienylacetyl chloride (1.31 S * 0.0815 moles) and methylene chloride (6 mL) are heated at 40-45 ° C in an oil bath under nitrogen. 20 hours. The reaction mixture is poured into hexane (100 mL) and filtered. Removal of the solvent gives 7 - [(D-5-trichloroethoxycarbonylamino-5-carboxyvinyl) -2-thienylacetylamino] -3-acetoxymethyl-3-cephem-4-carboxylic acid dibenzhydryl ester.

Step D: 3-Acetoxymethyl-7- (2-thienyl-acetamido) -3-cephem-4-carboxylic acid benzhydryl ester 7 ° C (D-5-trichloro-ethoxycarbonyl-amino-5-carbo-valeryl) -2-thienyl-acetylamino) -3-acetoxy : Dibentohydryl ester of ethyl 3-cephem-4-carboxylic acid is dissolved in ethyl acetate (10 ml) and added to a mixture of 90% aqueous acetic acid (10 ml) and zinc dust (1.0 g). The mixture is stirred for two hours at room temperature. * The reaction mixture is filtered to remove zinc. The reaction mixture is washed successively with 2 portions of water, cold sodium hydrogen carbonate solution and then saturated sodium chloride solution (15 → 0 ml). The ethyl acetate solution is dried over sodium sulfate, filtered and the solvent removed to give 1.9 S of crude product which is chromatographed on silica gel using a mixture of chloroform and ethyl acetate (50: 1) as eluent to give 0.380 g of product, m.p. after recrystallization from ethyl acetate. , 5-143 ° C.

VT: (CH 2 OH) ν max 263 ε 7580 Elemental analysis for C 29 H 26 N 2 O 2 S 2: Calculated: C, 61.91; H, 4.66; K, 4.98;

Found: C, 62.14; Ή, 4.84; li, 4.91.

Step E: 3- (Acetoxymethyl) -7- (2-thienylacetamido) -3-cephem-4-carboxylic acid

A cold solution of 3-acetoxymethyl-7- (2-thienylacetamido) -3-cofem-4-carboxylic acid benzhydryl ester (100 mg) in anisole (1.0 mL) and trifluoroacetic acid (0.5 mL) was stirred at 0 ° C. 35 minutes. Carbon tetrachloride (50 ml) is added and the reaction mixture is evaporated to dryness. The residue is triturated in hexane. The hexane is removed by decantation and this residue is dissolved in ethyl acetate (10 ml), concentrated to 1 ml and diethyl ether is added to cause precipitation. This precipitate is recrystallized from a mixture of diethyl ether and ethyl acetate to give 0.025 g of 3 '(acetoxymethyl) -7' (2-thienylacetamido) -3-sphera-4-carboxylic acid, m.p. 164 ° C. The melting point with the authentic sample was 163 ° C.

Example 5

Sodium 7- (2-thienylacetamido) -7-methoxy-3'-carbamoyl oxymethyl 3-cephem-4-carboxylate_

Step A: 78- (D-5-Benzoylamino-5-carboxy-valeramido) -3-carbamoyloxymethyl-7-methoxy-3 ", 4-carboxylic acid, disodium salt in 500 ml of an aqueous solution containing 48.5 mmol of monosodium 7β- (D-5 'amino-5-carboxivaleramido) -3-carbamoyloxymethyl-7-methoxy-3-cephem-4-carboxylate are added to 50% sodium hydroxide to a pH of 9.5 · To this solution is added 15 ml [128 mmol] of benzoyl chloride with vigorous stirring. The pH is maintained at 9.5 ° C for 30 minutes by adding sodium hydroxide as needed. The pH of the solution is then adjusted to 4.0 with concentrated hydrochloric acid, and washed twice with ethyl acetate.

14 6021 0

The aqueous layer is cooled to 0 ° C, and with stirring 200 ml of isopropanol and 300 ml of ethyl acetate are added, the pH is adjusted to 2.0 with hydrochloric acid. The organic layer is separated and the aqueous layer is re-extracted three times with ethyl acetate. The combined extracts are washed with brine, dried over sodium sulfate and concentrated in vacuo to give 43.0 g of a dark oil. The oil is dissolved in 200 any ethanol and a solution of 30 g of the sodium salt of 2-ethylhexeanoyl acid is added. The slurry is cooled to 0 ° C, filtered, washed with ethanol and dried in vacuo to give 28.8 g of [102 56] disodium 7β- (D-5-t-benzoylamino-5-carboxyvaleramido) -3-carbamoyloxymethyl- 7-Methoxy-3 'cephem-4-carboxylate, which is 67% pure compared to a chromatographically pure standard sample.

Step B: 7β- (D “5“ Benzoylamino-5-carboxivaleramido) -3-carbamoyloxymethyl-7-methoxy-3- * cephem-4-carboxylic acid dimethoxy methyl ester

To a slurry of 20 g of disodium 7β- (D-5-benzoylamino-5 ** carboxysivaleramido) -5-carbamoyloxymethyl-7-methoxy-3-cephem-4-carboxylate in 200 ml of acetonitrile at 0 ° C is added by dropwise addition of 16 ml of 6-mol chloromethyl methyl ether over 90 minutes. After one hour of addition, 6 ml of S-collidine was added. The slurry is stirred for an additional 2 hours at 0 ° C. The mixture is then diluted with 500 ml of methylene chloride and washed twice with dilute phosphoric acid, once with dilute sodium bicarbonate and once with 5% sodium chloride. The aqueous layers are washed back with 30 ml of methylene chloride. The organic phase is dried over sodium sulfate and concentrated in vacuo to about 100 ml.

The solution is passed through a 200 ml layer of Silica Gel G, washed with 200 ml of methylene chloride, then eluted with 800 ml of ethyl acetate. The ethyl acetate eluates are concentrated in vacuo to give 18.5 S of a yellow oil.

The crude product is recrystallized from 50 ml of ethyl acetate to give 10.0 g of (67%) 7β- (D-5-benzoylamino-5-carboxivaleramido) -3-carbamoyloxymethyl-7-methoxy-3-cephem-4-carboxylic acid. dimethoxymethyl ester.

False C: Sodium 7- (2-thienylacetamido) -7-methoxy-3-carbamoyloxymethyl 3-cephem-4-carboxylate_

Slurry of 192 mg (0.3 mmol) of 7β- (D-5-benzoylamino-5-carboxylamidoamido) -3-carbamoyloxymethyl-7-methoxy-3-cephem-4-carboxylic acid diraethoxymethyl ester, 225 mg of powdered Linden Type 4A molecular sieve with 10 to 2 ° /> water, 0.3 ml of 2-thienylacetyl chloride and 4 ml of 15 6021 0 dichloroethane are boiled with vigorous stirring for 4 hours. The mixture is cooled to 65 [deg.] C. and 10 ml of 0 (05 moles of dichloroethane solution of t-butanol) are added over 2 hours, followed by heating for a further hour at 65 [deg.] C. The reaction mixture is cooled to 0 [deg.] C., filtered and washed with 5 ml. The filtrate is cooled to 0 [deg.] C. with stirring, and then 1.4 ml of a 1: 1 solution of hydrochloric acid and methanol are added, and the resulting mixture is stirred at about 15 [deg.] C. for 3 hours, poured into 10 ml of water. with 1.6 g of sodium hydrogen carbonate The organic phases are discarded and the yield of sodium 7- (2-thienylacetamido) -7-methoxy-3-carbamoyloxymethyl-3-cephem-4-carboxylate from the aqueous layers is 659 ·

Step D: In Step A, p-chlorobenzoyl chloride, p-nitrobenzoyl chloride, or toluyl chloride is used in a similar manner. The maximum yield of sodium 7- (2-thienylacetamido) -7-methoxy-3-carbamoyloxymethyl-3-cephem-4-carboxylate of the desired product is 70, 6, 68 and 72, respectively.

Example 6

Hatium 7- (2-thienylacetamido) -7-methoxy-3-carbamoyloxymethyl 3-cephem-4-carboxylate_

A mixture of 2.76 g (4-mmol) of 7β- (D-5-tosylamino-5-carboxy-valeramido) -3-carbamoyloxymethyl-7H-methoxy-3-cephem-4-carboxylate dimethoxyethyl ester, 3 S dry A Linde-type 4A molecular sieve with less than 2 fo of water in 2 ml of thienylacetyl chloride (16 mmol) in 34 ml of dichloroethane is stirred at reflux for 5 hours. T-butanol, 0.38 ml (4 mmol) is then added and stirring is continued for 2 hours. After this time, a further 0.095 ml (1 mmol) of t-butanol are added and the reaction mixture is stirred at reflux for a further 1/2 hour. The reaction mixture is cooled to 0-5 ° 0 in an ice-water bath. The molecular sieve is removed by suction filtration and then washed with 40 ml of ice-cold methanol. The filtrate and washings are combined and cooled to 0 ° C. An ice-cold solution of 8.3 ml of concentrated hydrochloric acid and 9 * 5 ml of MeOH is added and the solution is warmed to 15 ° C and stirred at 15 ° C for 2 hours 40 minutes. After completion of the hydrolysis, the reaction is stopped by adding the mixture to a suspension of 22 g of sodium hydrogen carbonate in 120 ml of water at 0-5 ° C. The biphasic solution is stirred for 10 minutes. The heavy salt precipitate formed is removed by filtration and kept with a small amount of 5% NaCl solution containing 0.5 5% sodium hydrogen carbonate. The dichloroethane layer is separated and extracted into 2 x 20 ml of a solution of 0.5 N NaHCO 3 and 5 N NaCl. The aqueous fractions are combined and washed with 20 ml of dichloroethane.

16 6021 0

Liquid chromatographic analysis showed that the hydrogen carbonate solution contained 73% sodium 7- (2-thienylacetamido) -7-methoxy-3-carbamoyloxymethyl-3-cephem-4-carboxylate and 2.1% unchanged starting material.

Examples 7-20

Sodium 7- (2-thienylacetamido) -7 * (- methoxy-3-carbamoyloxymethyl-3-cephem-4-carboxylate)

Following the general procedure outlined in Example 6, dimethoxymethyl ester of 4 mmol (2.8 g) of 7/0- (D-5-tosylamino-5-carboxivaleramido) -3-carbamoyloxymethyl-7 '(- methoxy-3-cephem-4-carboxylate) allowed to react with the amounts of reagents and reaction conditions listed in the following table.The total reaction time in each case is about 16 hours.In each case, the molecular sieves used were Linde-type 4A heated to 700 ° C before the reaction, with a weight loss during drying of about 3 The screens are estimated to be essentially anhydrous with a water content of less than 2% by weight, with a reaction temperature of 67 ° C in each case.

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18 6021 0

Examples 21-25 The same general procedure as in Examples 6-20 is used, but the reaction temperature is varied according to the values in the table. The reaction mixture contains 4 mmol (2.8 g) of ββ- (D-5-tosylamino-5-carboxivaleramido) -3-carbamoyloxymethyl-7- (4-methoxy-3-cephem-4-carboxylate dimethoxymethyl ester, 3 g of dried Linde-type 4A sieve (less than 2% by weight of water) and 16 mmol of thienylacetyl chloride in 34 ml of dichloroethane.

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In all cases, the final product is sodium 7- (2-thienylacetamido) -7-methoxy-3-carbamoyloxymethyl-3-cephem-4-carboxylate.

Example 26 7- (2-Thienylacetamido) -7-methoxy-3-carbamoyloxymethyl-3-cephem-4-carboxylic acid

One gram of sodium N-O- (D-5-amino-5-carboxivaleramido) -3-carbamoyloxymethyl-7-methoxy-3-cephem-4-carboxylate is placed in a reaction vessel containing 20 ml of dichloroethane. To this is added 1 g of Linde molecular sieve type 4A as a powder with 15% water and 8 mmol (1 ml) of thienylacetyl chloride. The slurry is heated at reflux for 8 hours. After filtration, the molecular sieve is washed with methanol. The methanolic washing liquid of the sieve is analyzed and contains the desired product, N- (2-thienylacetamido) -7β- (methoxy-3-carbamoyloxymethyl-3-cephem-4-carboxylic acid, in 25% yield based on plate and liquid chromatographic determination. The dichloroethane layer of the reaction mixture is extracted NaHCO 4 and plate and liquid chromatographic determinations show that it still contains 5% of the desired product, and plate chromatographic analysis of the organic layer shows that it contains mixed anhydride, and after removal of the solvent by evaporation, the mixed anhydride is hydrolyzed at 50%. in acetone-water (50 ml) in the presence of 10 mol% pyridine, and plate and liquid chromatographic analyzes show that a further 13% of the final product is formed as the free acid, for a total yield of 156 mg of acid, i.e. a yield of 23%.

Claims (2)

6021 0 23 Claim: Process for the preparation of N-acylamido-cephalosporanic acid ester compounds of the formula R 1 HH s R'N - ν '\ X-NY ^ -CH 2 A' (1) C00R "wherein R 1 is hydrogen or methoxy, A ' is carbamoyloxy or acetoxy, R 'is an acyl group of the formula OR -CH-C- 1 H wherein R 1 is phenyl, 2-thienyl or 2-furyl, and R "is an easily removable ester protecting group such as methoxymethyl or benzhydryl , characterized in that a compound of formula R 1 H: C00R "wherein B is a protected aminoadipoyl group and R 1, R" and A 'are as defined above is reacted in an neutral solvent with an acylating agent of formula 0 R-CH -CCl2 H wherein R1 is as defined above, in the presence of a molecular sieve catalyst of synthetic aluminosilicate zeolite species 3A, 4A, 5A or 13X, having a pore size of about 3-15 Å, anhydrous or containing up to 30% by weight of hydrated water. %, and 6021 0 that acylation reaction then group B is cleaved to give a product of formula (I) either by prolonging the reaction time or by adding a cleavage agent such as benzyl alcohol, alkanol, lower alkylthiol or hydrochloric acid, or when the molecular sieve contains water of hydration acts as a hydrate water cleavage agent. 6021 0 For the preparation of β-3-acylamido-cephalosporan-cyanoester compounds of formula R1 H | II ς »I Γ - \ ^ \ m J> —cv COOR" color R ^ is a substance or methoxy, A 'is carbamoyloxy or acetoxy, R' is an acyl group with the formula OM R.-CH-C- 2-phenyl, 2-thienyl or 2-furyl, and R "are enantiomers of the ester group, are methoxymethyl or benzhydryl, converted to form, 1-H BN-rf Ί AN -CH2A ' 0 I COOR "color B is an amino acid group and R1, R" and A 'are selected from the group consisting of amino acids of the same formula as R o CH-CC1. in the form of a molecular sieve catalyst with a synthetic aluminosilicate zeolite of nitrogen type 3A, 4A, 5A or 13X, with a powder content of about 3 to 15 A and a water or a hydrogen hydrate of up to 30% by weight, and att Gruppen B