LT3871B - Additive crystalline salts of pure diastereomers of 1-cefem-4-carboxylic acid 1-(2,2-dimethyl-propionyloxy)ethyl esters, process for the preparation thereof, pharmaceutical compositions and process for the preparation thereof - Google Patents

Abstract

Invention is intended for effective treatment against bacterial infections. Additive crystal salts of both 3-cephem-4-carboxyl acid 1-/2.2-dimethylpropionyloxyl/-ethyl ester diastereomer acids, the common formula II of which is<IMAGE>(II) where X is a physiologically proper monobasic or multibasic inorganic or organic acid anion, δ=N-OH group is in sin-position for pharmaceutical compositions that have the derivatives, for the production methods of cephem derivatives also for the use of the cephem derivatives to fight bacterial infections.

Description

The present invention relates to crystalline, intestinal resorbable Ί- (2- (2-aminothiazol-4-yl) -2- (Z) -oximino-acetamido) -3-methoxymethyl-3-cephem-4-carboxylic acid 1- (2,2-dimethylpropionyloxy) for salts of the diastereomers of ethyl ester (formula I) h ₂ n

ND II ^z Y

NOH: onh / ⁵

V x ^{2 3}

CO ₂ CHOCC (CH ₃ ) ₃ [I] ^H 3 ^C and their preparation.

VFR patent application P 38 04 841 (European Patent AO-329 008) describes 7- (2- (2-aminothiazol-4-yl) -2- (Z) oximinoacetamido) -3-methoxymethyl-3-cephem-4 -Carboxylic acid esters. Of particular interest is the ester of Formula I as it is well absorbed in the gut of various species of animals and, following resorption by autogenous enzymes, is rapidly and completely degraded to the antibiotic active cephalosporin, which has a free carboxyl group.

VFR patent application P 39 19 259 (European patent A-O-402 806) describes stoichiometric crystalline salts of the ester of formula I and sulfuric acids which, being very stable, have more advantages than the free bases of formula I.

The ester of formula I has an asymmetric carbon atom at the 1-position of the ethyl group of the ester. The salts described in VFR patent application P 39 19 259 are mixtures of diastereomers.

Similar mixtures of diastereomers are present in the case of cefothiam-hexethyl, cefuroxime-acetyl, cefpodoxime-proxoxyl and DVE 28271.

Previous studies suggesting that this mechanism of intestinal resorption of cephem esters has no effect on intestinal resorption by stereochemistry at the 1-position of the ester ethyl group. In the case of Cefotiam-hexethyl diastereomers, this was experimentally determined by [T. Nishimuro et al., The Journal of Antibiotic Vol. XI (1987), 81-90].

Therefore, it was very unexpected when it was found that the resorption of the diastereomeric salts of the formula I in the intestine differs significantly, i.e. the better resorbable diastereomer exhibits a higher bioavailability than the mixture of diastereomers described in VFR patent application P 39 19 259.

Therefore, the present invention relates to diastereometrically pure salts of formula II in which the C = N-OH group is in syndrome. Of the two diastereomers with the [IS] configuration in the ester group, the less polar, with a higher bioavailability, the mayor of diastereo25 is more suitable.

N-C — CONH r

H II II I

HN ^ZX S ^{/ NOH}

C ») ^ h ₂ och ₃

I x

CO ₂ CHOCC (CH ₃ ) ₃ > fHX [uj »3 ^C

In the general formula II, HX is a monobasic or polybasic acid, wherein X may be an inorganic or organic physiologically acceptable anion.

For inorganic acid, HX is, for example, stoichiometric HCl, HBr, HJ, HBF, HNO ₃ , HClO ₄ , H _{2 5} O ₄ or

Quantity of H ₃ PO ₄ . In the case of organic acid, HX is an aliphatic or aromatic sulfuric acid. Inorganic acids include HCl, HBr, and H ₂ SO ₄ , as well as organic acids such as methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid and 4-ethylbenzenesulfonic acid. Particularly noteworthy are benzenesulfonic acid, p-toluenesulfonic acid and 4-ethylbenzenesulfonic acid.

The present invention also relates to processes for the preparation of diastereomeric compounds of formula II, which differ in that

1) The compound of formula II is obtained:

II

N

CO ₂ CHOCC (CH ₃ ) ₃ [III]

chromatographic separation of the diastereomers, separation of the trityl groups and preparation of the acid addition products, or

(2) crystallization from the mixture of diastereomers of formula II yielding a less polar diastereomer, or

3) Intermediate of formula IV in the form of the diastereomers separated:

π ₂ ν / ^s x

CO ₂ CHOCC (CH ₃ ) ₃ [IV] ^H 3 ^c and they are converted into the diastereomers of the formula II.

The preparation of a mixture of diastereomers of formula III required by method 1 is described in VFR patent application P 38 04 841.

The diastereomers are separated by chromatography on silica gel using toluene and ethyl acetate as solvents. The ratio of toluene to ethyl acetate can vary within a wide range from 3: 1 to 20: 1, preferably from 10: 1 to 15: 1. 20-80 parts of silica gel are used to distribute one part of the mixture, most preferably 3050 parts.

The pure diastereomers of formula III thus obtained are converted into the salts of formula II using the methods already described in VFR patents P 38 04 841 and P 39 19 259 for mixtures of diastereomers.

The less polar diastereomer of Formula II is obtained by Method 2 by crystallization of mixtures of diastereomers from organic solvents.

Under normal conditions of crystallization, the substance is dissolved in a solvent by heating to reflux. The compounds of formulas I and II are degraded under these conditions. However, salts may be recrystallized according to the method described below.

One portion of the mixture of diastereomers is initially dissolved in 1-5 parts of an organic solvent, preferably 1-2 parts, for example dimethylformamide or dimethylacetamide. The resulting solution is added dropwise to 5 to 50 times the volume of the organic solvent (e.g., alcohol, ester, ether, ketone, nitrile) such as methanol, ethanol, n-propanol, isopropanol, n-butanol, tert-butanol, ethyl acetate, butyl acetate, acetone, diethyl ether, diisopropyl ether and acetonitrile. It is expedient to use 10-20 times the volume of n-propanol, isopropanol and n-butanol.

The drip may last from 10 minutes. up to 2 hours, most appropriate from 30 minutes up to 1 hour The crystallization is further stirred for 1-18 hours, preferably 3-6 hours. The temperature should be maintained at 0-40 ° C, preferably at 20-25 ° C.

The resulting salts are isolated by conventional laboratory methods, such as filtration, and remove traces of solvents by drying under high vacuum (<1 Tor), such as phosphorus pentoxide.

Repeated use of the process described above yields a pure diastereomer of formula II (HX = p-toluenesulfonic acid) with high intestinal resorption.

The compound of formula IV obtained by method 3 in the form of a mixture of diastereomers is described in VFR patent P 38 04 841.

Diastereomers may be resolved by crystallization of salts of formula V:

O '

I-N 1 -CH "OCH" X ^{2 3}

CO ₂ CHOCC (CH ₃ ) ₃ ♦ HY [V] »

The HY of formula V is a single or polybasic acid, and Y may be an inorganic or organic anion.

In the case of an inorganic acid, HY may be, for example, HCl, HBr, HJ, HNO ₃ , HClO ₄ , HSCN, H ₂ SO ₄ or H ₃ PO ₄ . In the case of organic acids, HY may be aliphatic or aromatic sulfuric, carboxylic and phosphonic acids. For example, the following organic acids may be used: benzenesulphonic acid, p-toluenesulphonic acid, 4-ethylbenzenesulphonic acid, 4-bromobenzenesulphonic acid, 2-mesitylensulphonic acid, 4-diphenylsulphonic acid, naphthalene-1,5-disulphuric acid, .

The following acidic components may be preferred: HCl, HBr, Benzenesulphonic acid, p-toluenesulphonic acid,

4-ethylbenzenesulfonic acid and 4-diphenylsulfonic acid.

The salt of formula V is obtained by mixing solutions of a mixture of diastereomers of formula IV and an acidic HY component. Organic solvents used are, for example, esters and ethers, alcohols, ketones, hydrocarbons, nitriles and halogenated hydrocarbons and mixtures thereof. Preference is given, for example, to benzene, toluene, ethyl acetate, butyl acetate, methanol, ethanol, n-propanol, isopropanol, tert-butanol, diisopropyl ether, acetonitrile, dichloromethane, acetone and mixtures thereof.

In the case of inorganic acids, if the organic solvent is soluble in water, water may also be used as the solvent. Solutions of HCl and HBr in organic solvents are obtained, for example, by the addition of hydrogen chloride or hydrogen bromide, or of acetyl halides, phosphorus halides and phosphorus oxyhalides and alcohol (halo = Cl, Br).

The ratio of formula IV to the acidic component is important to increase the yield of one of the diastereomers. An equivalent of 0.22.0, most preferably 0.3-1.0, of the acidic component is used per equivalent of a mixture of diastereomers.

The acidic component is added at room temperature. Further complete stirring is continued for up to 10 hours, depending on the acidic component and the solvent. If necessary, complete precipitation is cooled from room temperature to -78 ° C.

The salts obtained after filtration, if necessary, are purified by crystallization. For this purpose, the solvents and mixtures thereof described above are used. The choice of the optimal solvent depends on the acidic component used. For example, methanol, ethanol, n-propanol and isopropanol are suitable salts of p-toluenesulfonic acid.

The method differs in that the precipitation of the diastereomers of formula IV is carried out in two successive partial steps. For example, combining a solution of a mixture of formula IV diastereomers with a solution of the acidic HY component first precipitates and separates by filtration the less soluble Form V diastereomer, and after filtration precipitates the more soluble Form V diastereomer. In successive partial steps, the acidic component HY may be the same or different, and the different acidic HY components may be added in any order. For example, when the acidic HY component is correctly selected, a more polar diastereomer of Formula IV or a more non-polar diastereomer may be initially precipitated in the form of a less soluble salt.

When the acidic component is selected, both diastereomers of formula V can be obtained in pure form. For example, hydrochloric acid or hydrobromic acid gives a more polar diastereomer, while benzoic acid, 4-ethylbenzenesulfonic acid, biphenylsulfonic acid or p-toluenesulfonic acid yields a less polar diastereomer.

Similarly, mixtures of diastereomers of formula IV may be obtained from compounds of formula VI.

R ¹ HN 15

CO CHOCC (CH_) ₀

I II ^{33 H} co [VI]

In this formula, R ¹ is an amino protecting group commonly used in peptide chemistry, such as formyl, tert-butoxycarbonyl, phenoxyacetyl, phenylacetyl, allyloxycarbonyl, benzyloxycarbonyl and 4-nitrobenzyloxycarbonyl.

The separation of the protecting groups takes place in known ways.

For example, the formyl group and the tert-butyloxycarbonyl group can be cleaved by acid. The phenoxyacetyl group and the phenylacetyl group can be cleaved by phosphorus pentachloride or by the enzyme penicillinacylase. In the case of the allyloxycarbonyl group, the separation is carried out with Pd [P (C ₆ H ₅ ) ₃ ] ₄ . The benzyloxycarbonyl group and the 4-nitrobenzyloxycarbonyl group can be hydrolytically removed.

Cleavage of the phenoxyacetyl group or the phenylacetyl group 35 with the aid of phosphorus pentachloride in a continuous process yields larger amounts of the more polar diastereomeric hydrochloride; untreated mixed phosphoric acid esters - chloro anhydrides, which slowly release hydrochloric acid - are involved in the treatment.

Diastereometrically pure compounds of formula V can be obtained from compounds of Formula VI by first separating the diastereomers and then cleaving the protecting group. The diastereomers can be separated by crystallization or chromatography, and the conditions are chosen depending on the protecting group R ¹ . For example, when R ¹ is a phenoxyacetyl group, the diastereomers may be separated by chromatography on silica gel using a mixture of organic solvents.

—N,

[VII]

CO ₂ CHOCC (CH ₃ ) ₃ ^h 3c o

Another alternative for the preparation of pure diastereomers of formula IV follows from Schiff's bases of formula VII wherein R ² is a phenyl or naphthyl residue which may contain (C ₁ -C ₄ ) alkyl, phenyl, methoxy, halogen (e.g. fluorine, bromine, chlorine, iodine). ) or a nitro group as substituents.

The mixture of diastereomers of Schiff's bases of formula VII is separated either by chromatography on silica gel or by fractional crystallization. Schiff's reversible cleavage to pure diastereomers of Formula IV is accomplished by known methods, for example, by acid hydrolysis or Girard-T-reagent.

(Diastereomeric bases of formula IV obtained from diastereomerically pure salts of formula V by the known methods (mean value of tests)

67.7%

19.7%: 1) 39.2% is converted to the diastereomeric salts of formula II as described in VFR patents P 38 04 841 and P 39 19 259.

The importance of the present invention is that the less polar diastereomer of Formula II is more resorbable in the gut. This is illustrated by the salt data of p-toluenesulfonic acid in Table 1.

table

Composition of Diastereomers Diastereomer of Example 1 Diastereomer of Example 1 Mixture of Diastereomers (Ratio 1

The table shows (0.24 h) the amount of 7- (2- (2-aminothiazol-4-4-yl) -2- (Z) -oximino-acetamido) -3-methoxymethyl-3-cephem-4-carboxylic acid found. in dog urine (n = 4) following oral administration of 1- (2,2-dimethyl-propionyloxy) -ethyl ester 7- (2- (2-aminothiazol-4-yl) -2- (Z) -oximino-acetamide) - 3-methoxymethyl-3-cephem-4-carboxylic acid p-toluenesulfonate (dose: 10 mg / kg based on biologically active antibacterial agent). The amount of biologically active substance in the urine is determined microbiologically by diffusion on an agar assay using Mueller - Hinton agar (with 10% sheep blood) and Streptococcus pyogenes A 77 as a test chroma organism.

The compounds of the formula II according to the invention are administered orally in the form of conventional pharmaceutical compositions such as capsules, tablets, powders, syrups or suspensions. The dose depends on the age, symptoms and weight of the patient as well as the duration of treatment. As a rule, however, it is from about 0.1 g to about 5 g per day, preferably about 0.2 g to 3 g per day.

Most preferably, the compounds are administered in divided doses, for example 2 to 4 times per day, so that a single dose may be, for example, 50 to 500 mg of the biologically active substance.

Oral compositions may contain conventional carriers and / or diluents. For example, for capsules or tablets, binders such as gelatin, sorbitol, polyvinylpyrrolidone or carboxymethylcellulose, diluents such as lactose, sugar, starch, calcium phosphate or polyethylene glycol, lubricating agents such as talc or magnesium stearate are used. Liquid compositions such as aqueous or oily suspensions are suitable for syrups or conventional forms of known composition.

The following examples illustrate the compounds of formula I according to the invention 7- (2- (2-aminothiazol-4-yl) -2- (Z) -oximino-acetamido) -3-methoxy-methyl-3-cephem-4-carboxylic acid 1,1 The preparation of diastereomerically pure salts of - (2,2-dimethoxypropionyloxy) -ethyl esters further illustrates the invention but does not limit its scope.

example

Preparatory stage

2- (2-Tritylaminothiazol-4-yl) -2- (Z) -trityloxyimino-acetic acid chloro anhydride (trityl = triphenylmethyl) to 42.0 g (54 mmol) of 2- (-2-tritylaminothiazol-4-yl) -2 - (Z) Triethylammonium salt of trityloxyiminoacetic acid in 400 ml of anhydrous methylene chloride solution - 11.4 g (55 mmol) of phosphorus pentachloride in 200 ml of anhydrous methylene chloride are added dropwise over 30 minutes at a temperature of -50 ° C. C. After 60 minutes at -70 ° C, the solvent is evaporated off under vacuum (bath temperature should not exceed 30 ° C). It is then briefly dried in a deep vacuum. The crude product thus obtained is dissolved in 100 ml of anhydrous methylene chloride and used for acylation.

Stage

1- (2,2-Dimethylpropionyloxy) -ethyl 3-methoxymethyl-7- (2- (2-tritylaminothiazol-4-yl) -2- (Z) trityloxyiminoacetamido) -3-cephem-4-carboxylic acid

To a suspension of 14.0 g (57 mmol) of 7-amino-3-methoxymethyl-3-cephem-4-carboxylic acid in 160 mL of anhydrous methylene chloride was added 9.5 mL (64 mmol) of DBV at 0 ° C and stirred for a further 30 min at 0 ° C. At C. 20.8 g (81 mmol) of 1-iodethyl ester of 2,2-dimethylpropionic acid are then added, the mixture is stirred for another 30 min at 0 ° C and allowed to warm to room temperature over 30 min. After cooling to 0 ° C, pure 2- (2-tritylaminothiazol-4-yl) -2- (Z) trityloxyiminoacetic acid, chloro anhydride (approximately 54 mmol) was added dropwise, dissolved in 100 ml of methylene chloride and stirred at 2 or 0 ° C. The reaction mixture was concentrated in vacuo. The residue is dissolved in ethyl acetate and the solution is washed several times with 5% sodium thiosulphate solution, followed by saturated sodium bicarbonate solution and saturated sodium chloride solution. The organic phase is dried over sodium sulfate and evaporated to dryness in vacuo. The product is purified by chromatography on silica gel (35-70µμ) (column: 50 cm x 8.5 cm, toluene / ethyl acetate = 5/1). Yield: 36.5 (66%). Diastereomeric ratio 1/1.

Stage

Chromatographic separation of diastereomers g of diastereomeric mixture is chromatographed on silica gel (35-70 µm, column: 46 x 7.5 cm) using a toluene / ethyl acetate (15/1) mixture at a flow rate of 50 ml / min. Concentration in vacuo gives 6.0 g of a non-polar diastereomer 1 and 4.5 g of a polar 2 diastereomer.

Diastereomer

R f (toluene: ethyl acetate 5: 1) = 0.48 ^{X 1} H-NMR (DMSO - d ₆ , 270 MHz): δ = 1.15 (singlet,

9H, C (CH ₃ ) ₃ ), 1.50 (doublet 3H, OCH (CH ₃ ) O), 3.20 (singlet, 3H, OCH ₃ ), 3.57 (AB-system, 2H, SCH ₂ ). , 4.5 (singlet, 2H, CH ₂ O), 5.25 (doublet, 1H, H-6), 5.89 (double doublet, 1H, H-7), 6.59 (singlet, 1H, thiazole) -H), 6.89 (quadruple, 1H, OCH (CH ₃ ) O), 7.12-7.37 (multiplet, 3 OH, aromatic H), 8.75 (singlet 1H, NH), 9.90 (doublet, 1H, amide NH).

Diastereomer

R f (toluene: ethyl acetate 5: 1) = 0.40 ^X H - NMR (CDCl ₃ , 270 MHz): δ 1.22 (singlet, 9H,

C (CH ₃ ) ₃ ), 1.56 (doublet, 3H, OCH (CH ₃ ) O), 3.30 (singlet, 3H, OCH ₃ ), 3.39 (AB system, 2H, SCH ₂ ), 4 , 27 (singlet, 2H, CH ₂ O), 5.05 (doublet, 1H, H-6), 6.04 (double doublet, 1H, H-7), 6.41 (singlet, 1H, thiazole-H). ), 6.75 (singlet, 1H, NH), 7.04 (quadruple, 1H, OCH (CH ₃ ) O), 7.10 - 7.44 (multiplet, 30H, aromatic H).

Stage

7- (2- (2-Aminothiazol-4-yl) -2- (Z) -oximinoacetamido) -3-methoxymethyl-3-cephem-4-carboxylic acid 1- (2,2-dimethylpropionyloxy) -ethyl ester

Diastereomer

A solution of 6.0 g (5.9 mmol) of diastereomer 1 from Step 2 in 15 mL of formic acid is added dropwise to 3 mL of water. Stir 90 min at room temperature, then 30 min at 0 ° C. The crystallized triphenylcarbinol is filtered off and washed with a small amount of formic acid (1/5) diluted with water. The filtrates are mixed, 60 ml of ethyl acetate and 30 ml of water are added and the solution is adjusted to pH 3.0 with ice-cold bath with 2 N sodium hydroxide solution. The organic layer is separated, washed twice with 50 ml of water and again mixed with 50 ml of water. Add 40% sodium hydroxide solution to pH = 6.5 (solution temperature should not exceed 10 ° C). The organic layer was separated, washed with a saturated sodium chloride solution, dried over sodium sulfate and concentrated in vacuo to a quarter of the original volume. The solution thus obtained is added dropwise to 150 ml of diisopropyl ether. After stirring for 60 min at room temperature, the product is suctioned off, washed with diisopropyl ether, air dried for 18 hours and then vacuum dried over phosphorus pentoxide. Yield: 1.7 g (54%).

¹ H-NMR (DMSO - d ₆ , 270 MHz): δ = 1.15 (singlet, (9H, C (CH ₃ ) ₃ ), 1.48 (doublet, 3H, OCH (CH ₃ ) O, 3, 20 (singlet, 3H, OCH ₃ ), 3.55 (AB - system, 2H, SCH ₂ ), 4.13 (singlet, 2H, CH ₂ O), 5.21 (doublet, 1H, H-6), 5.85 (doublet of doublets, 1H, H-7), 6.65 (singlet, 1H, thiazole-H), 6.87 (quadruple, 1H, OCH (CH ₃ O), 7.11 (singlet, 2H, NH ₂ ), 9.47 (doublet, 1H, amide NH),

11.28 (singlet, 1H, NOH).

Diastereomer

4.5 g (44 mmol) of diastereomer 2 obtained in Step 2 are treated in the same manner. Yield: 1.7 g (71%).

¹ H-NMR (DMSO - d ₆ , 270 MHz): δ = 1.16 (singlet, 9H, C (CH ₃ ) ₃ ), 1.49 (doublet, 3H, OCH (CH ₃ ) O), 3, 20 (singlet, 3H, OCH ₃ ), 3.55 (AB system, 2H, SCH ₂ ), 4.12 (singlet, 2H, CH ₂ O), 5.19 (doublet, 1H, H-6), 5 , 82 (double doublet, 1H, H-7), 6.66 singlet, 1H, thiazole-H), 6.93 (quadruple, 1H, OCH (CH ₃ ) O), 7.10 (singlet, 2H, NH). ₂ ), 9.45 (doublet, 1H, amide NH), 11.29 (singlet, 1H, NOH).

Stage

7- (2- (2-Aminothiazol-4-yl) -2- (Z) -oximinoacetamido) -3-methoxymethyl-3-cephem-4-carboxylic acid 1- (2,2-dimethylpropionyloxy) -ethyl ester p-toluenesulfonate

Diastereomer g A suspension of 1 diastereomer g (1.85 mmol) of Step 3 in 35 mL of n-propanol is mixed with 383 mg (2.0 mmol) of p-toluenesulfonic acid monohydrate in 1 mL of n-propanol. The solid dissolves, and after a few minutes the salt begins to crystallize. After stirring for 1 additional hour at room temperature, the product is filtered off and washed with 5 ml of n-propanol and 10 ml of diisopropyl ether. Dry for 18 hours in air, then in a deep vacuum over calcium chloride and paraffin. Yield: 1.09 g (83%).

(α) = +48.8 (c = 1, methanol). Lyd. temp. > 200 ° C (dec.).

^{Τ Η} - NMR (DMSO - d _6, 270 MHz): δ = 1.15 (singlet, 9H, C (CH _{3) 3),} 1.48 (doublet, 3H, OCH (CH ₃₎ O) 2, 29 (single16 th, 3H, aryl-CH ₃ ), 3.20 (singlet, 3H, OCH), 3.59 (AB - system, 2H, SCH ₂ ), 4.14 (singlet, 2H, CH ₂ O). ,

5.24 (doublet, 1H, H-6), 5.85 (double doublet, 1H, H-7), 6.82 singlet, 1H, thiazole-H), 6.87 (quadruple, 1H, OCH (CH). ₃ ) O, 7.08-7.15 and 7.45-7.52 (2x multiplet, 2 χ 2H, aromatic H), 8.0-8.8 (broad,

3H, NH), 9.67 (doublet, 1H, amide NH), 12.04 (singlet, 1H, NOH).

Diastereomer

1.6 g (2.5 mmol) of diastereomer 2 obtained in Step 3 is crystallized from 15 ml of n-propanol to give the p-toluene 2 sulphonic acid salt. Yield: 1.4 g (66%) (a) _D = + 12.7 (c = 1, methanol). Lyd. temp. > 200 ° C (dec.).

¹ H - NMR (DMSO - d ₆ , 270 MHz): δ = 1.17 (singlet, 9H, C (CH ₃ ) ₃ ), 1.49 (doublet, 3H, OCH (CH ₃ ) O), 2, 29 (singlet, 3H, aryl-CH ₃ ), 3.21 (singlet, 3H, OCH ₃ ), 3.57 (AB-system, 2H, SCH ₂ ), 4.13 (singlet, 2H, CH ₂ O). , 5.22 (doublet, 1H, H-6), 5.82 (double doublet, 1H, H-7), 6.85 (singlet, 1H, thiazole-H), 6.94 (quadruple, 1H, OCH). (CH ₃ ) O, 7.08-7.16 and 7.45-7.52 (2x multiplet, χ 2H, aromatic H), 8.4-8.9 (broad, 3H, NH ₃ ),

9.68 (doublet, 1H, amide NH), 12.12 (singlet, 1H,

NOH).

An example

7- (2- (2-Aminothiazol-4-yl) -2- (Z) -oximinoacetamido) -3-methoxymethyl-3-cephem-4-carboxylic acid 1- (2,2-dimethylpropionyloxy) -ethyl ester p-toluenesulfonate (1) diastereomer g (70 mmol) 7- (2- (2-aminothiazol-4-yl) -2- (Z) -oximinoacetamido) -3-methoxymethyl-3-cephem-4-carboxylic acid 1- (2,2-dimethylpropionyloxy) of ethyl ester p-toLT 3871 B luensulfonate (diastereomer 1 / diastereomer 2 = 63/37) is slowly dissolved in 65 ml of dimethylacetamate by slow heating, and the solution is added dropwise to 450 ml of n-propanol over 1 hour or stirred at room temperature for 4 hours. The product is filtered off, washed successively with n-propanol and diisopropyl ether and dried in air first, then in vacuo over phosphorus pentoxide, yield: 29.5 g (59%), diastereomer 1 / diastereomer 2 = 79/21.

If necessary, three recrystallizations from 65 ml of dimethylacetamide and 450 ml of propanol yield 8 g (25%) of diastereomer 1 in greater than 97% purity.

The spectral data correspond to diastereomer 1 obtained according to the procedure described in Example 1. The diastereomeric ratio was determined by high performance liquid chromatography (Lichrospher 60, RP - select B, 125 x 4 mm, methanol / water = 5/6 with 0.12% ammonium dihydrophosphate, pH = 2.3, flow rate: 1 ml / min, detection at A = 228 nm, retention times: 1 diastereomer: 14.6 min diastereomer: 11.7 min).

An example

Stage

3-Methoxymethyl-7-phenoxyacetamido-3-cephem-4-carboxylic acid sodium salt

The sodium salt is derived from a carboxylic acid (Fujimoto et al., J. Antibiotic XL / 1987 / 370-84).

50.3 g (133 mmol) of carboxylic acid and 11.7 g (140 mmol) of sodium hydroxycarbonate are mixed with 900 ml of water. After filtration and drying, freezing yields the sodium salt. Yield: 47.8 g (67%).

¹ H-NMR (D ₂ O, 270 MHz): δ = 3.28 (singlet, 3H, OCH ₃ ),

3.42 (AB - system, 2H, SCH ₂ ), 4.16 (AB - system, 2H,

CH ₂ O) 4.72 (AB - System, 2H, OCH ₂ CO), 5.12 (doublet,

1H, H-6), 5.67 (doublet, 1H, H-7), 6.98-7.12, and 7,327.42 (2x multiplet, 5H, aromatic H).

Stage

1- (2,2-Dimethylpropionyloxy) ester of 3-methoxymethyl-7-phenoxyacetamido-3-cephem-4-carboxylic acid

42.8 g (107 mmol) of 3-methoxymethyl-7-phenoxyacetamide-3-cephem-4-carboxylic acid sodium salt dissolved in 430 ml of anhydrous dimethylformamide are mixed with 25.7 g (100 mmol) of 2,2-dimethylpropionic acid 1-iodethyl ester. The reaction mixture was stirred at room temperature for a further 1 hour, then poured into a mixture of 2.5 L of water and 1.5 L of ethyl acetate. The aqueous layer was extracted once more with ethyl acetate. The combined organic layers were washed with a saturated sodium chloride solution, dried over magnesium sulfate and evaporated in vacuo. Yield: 50.3 g (98%); Diastereomer 1 / Diastereomer 2 =

50/50.

Stage

Chromatographic separation of diastereomers The resulting mixture of diastereomers is separated by medium pressure chromatography (silica gel: 35-70 µm, 40 g silica gel, 1 g of material, toluene / ethyl acetate / diisopropyl ether = 120/15/6).

Diastereomer ^Χ Η - NMR (DMSO - d ₆ , 270 MHz): δ = 1.14 (singlet, 9H, C (CH ₃ ) ₃ ), 1.48 (doublet, 3H, OCH (CH ₃ ) O), 3 , 21 (3871 singLT B slow, 3H, OCH _3), 3.59 (AB system, 2H, SCH _2), 4.14 (singlet, 2H, CH ₂ O), 4.52 (doublet, 2H, OCH ₂ CO), 5.18 (doublet, 1H, H-6), 5.78 (double doublet, 1H, H-7), 6.87 (quadruple, 1H, OCH (CH ₃ ) O, 6.9-. 7.0 and 7.25-7.32 (2x multiplet, 5H, aromatic H), 9.13 (doublet,

1H, amido NH).

Diastereomer ^{X 1} H - NMR (DMSO - d ₆ , 270 MHz): δ = 1.17 (singlet, 9H, C (CH ₃ ) ₃ ), 1.49 (doublet, 3H, OCH (CH ₃ ) O), 3 22 (singlet, 3H, OCH _3), 3.60 (AB system, 2H, SCH _2), 4.13 (singlet, 2H, CH ₂ O), 4.52 (doublet, 2H, OCH ₂ CO) , 5.18 (doublet, 1H, H-6), 5.75 (double doublet, 1H, H-7), 6.90-6.99 (multiplet, 4H, aromatic H and OCH (CH ₃ ) O). , 7.20-7.32 (multiplet, 2H, aromatic H), 9.12 (doublet, 1H, amide NH).

Stage

7-Amino-3-methoxymethyl-3-cephem-4-carboxylic acid 1- (2,2-dimethylpropionyloxy) -ethyl ester

Diastereomer p-toluenesulfonate

A solution of 3.93 g (7.8 mmol) of the diastereomer 1 of stage 3 and 1.07 ml (8.45 mmol) of N, N-dimethylaniline in 39 ml of anhydrous methylene chloride is added dropwise at 1.40 g (9). , 32 mmol) of phosphorus pentachloride in 32 ml of anhydrous methylene chloride solution while maintaining the temperature of the solution at -25 ° C. Allow to stand for 2 hours until the temperature reaches -10 ° C, after which 19.4 ml of isobutanol are added immediately. After 10 minutes, the solution is poured into a mixture of 250 ml of saturated sodium hydroxycarbonate solution and 250 ml of ethyl acetate and the organic layer is separated as soon as possible. The aqueous layer was extracted once more with ethylLT 3871 B acetate. The combined organic layers are washed with a saturated sodium chloride solution, dried over magnesium sulfate and evaporated in vacuo. The residue is dissolved in 5 ml of ethyl acetate and mixed with a solution of 1.47 g (7.47 mmol) of p-toluenesulphonic acid monohydrate in 10 ml of ethyl acetate. The product is filtered, washed with ethyl acetate and dried over phosphorus pentoxide in vacuo. Yield: 2.57 g (61%).

¹ H - NMR (DMSO - d ₆ , 270 MHz): δ = 1.15 (singlet, 9H, C (CH ₃ ) ₃ ), 1.48 (doublet, 3H, OCH (CH ₃ ) O), 2, 29 (singlet, 3H, aryl-CH ₃ , 3.23 (singlet, 3H, OCH ₃ ), 3.69 (AB - system, 2H, SCH ₂ ), 4.16 singlet, 2H, CH ₂ O),

5.24 and 5.28 (2x doublet, 2 χ 1H, H-6 & H-7), 6.89 (quadruple, 1H, OCH (CH ₃ ) O), 7.12 (doublet, 2H, aromatic) H), 7.49 (doublet, 2H, aromatic H), 8.88 (singlet, 2H, NH ₂ ).

Diastereomer hydrochloride

506 mg (1 mmol) of the Phase 2 diastereomer is cleaved according to the procedure described above to give the phenoxyacetyl group. 2 The diastereomer crystallizes from ethyl acetate in the form of the hydrochloride. Yield: 223 mg (55%).

¹ H-NMR (DMSO - d ₆ , 270 MHz): δ = 1.17 (singlet, 9H, C (CH ₃ ) ₃ ), 1.49 (doublet, 3H, OCH (CH ₃ ) O), 3, 24 (singlet, 3H, OCH ₃ ), 3.63 (AB-system, 2H, SCH ₂ ), 4.20 (singlet, 2H, CH ₂ O), 5.21 (double doublet, 2 χ 1H, H- 6, and H-7), 6.93 (quadruple, 1H, OCH (CH ₃ ) O), 9.18 (singlet, 2H, NH ₂ ).

Stage

3-Methoxymethyl-7- (2 (- (2-tritylaminothiazol-4-yl) -2- (Z) triethyloxyiminoacetamido-3-cephem-4-carboxylic acid, 1- (2,2-dimethylpropionyloxy) -ethyl ester (1 diastereomer) )

1.5 g (2.75 mmol) of the 4-step 1 diastereomer tosylate are suspended in 100 mL of ethyl acetate and 30 mL of water. Adjust the pH to 6.5 with vigorous stirring at 0 ° C with saturated sodium bicarbonate solution. The organic layer is washed first with 30 ml of water, then with a saturated sodium chloride solution, dried over sodium sulphate and evaporated to dryness in vacuo. Yield: 1.03 g (98%).

According to the procedure described in Example 1, 1.81 g (2.3 mmol)

The triethylammonium salts of 2- (2-tritylaminothiazol-4-yl) -2- (Z) -trityloxyiminoacetic acid are converted into the acid anhydride.

To a solution of the ester in 880 mg (2.3 mmol) in 10 ml of anhydrous methylene chloride at -5 ° C is added dropwise acid 8 ml of anhydrous methylene chloride. After 2 hours, isolate as described in Example 1. The resulting product is chromatographed using a mixture of toluene and ethyl acetate (5/1) to give the diastereomerically pure product. Yield: 2.38 g (99%).

The spectral data are in agreement with the diastereomer 1 of Example 1. Further modifications are made in the manner described there.

An example

7-Amino-3-methoxymethyl-3-cephem-4-carbonic acid 1- (2,2-dimethylpropionyloxy) -ethyl ester, 1 diastereomer tosylate and 2 diastereomer hydrochloride

Step 3 of Example 3 describes the resolution of the phenoxyacetyl group using 3.03 g (6 mmol) of 3-methoxymethyl-7-phenoxyacetamide-3-cephem-4-carboxylic acid 1- (2,2-dimethylpropionyloxy) ethyl ester (diastereomer 1 / diastereomer 2). = 52/48). The dried organic layers are concentrated to a volume of 10 ml. After cooling to 0 ° C, the diastereomer 2 is precipitated in the form of the hydrochloride and filtered off. (Yield: 759 mg = 31%) 1.13 g (5.9 mmol)

The mother liquor is mixed with a solution of p-toluenesulfonic acid monohydrate in 5 ml of ethyl acetate. The precipitated salt of p-toluene sulfuric acid is filtered off, washed with a small amount of ethyl acetate and dried over phosphorus pentoxide in vacuo. Yield: 667 mg (23%).

According to high performance liquid chromatography 1, the diastereomer content is more than 97% (high performance liquid chromatography: Lichrospher 100 RP 18.5 µm, 125 x 4 mm, flow rate 1 ml / min, detection at A-254 nm, water / methanol 52/48 with 0.1% ammonium acetate, retention time: 1 diastereomer = 12.1 min, 2 diastereomer = 11.8 min).

Intermediates are converted to final according to the procedures described in the examples above.

An example

7-Amino-3-methoxymethyl-3-cephem-4-carbonic acid 1- (2,2-dimethylproponyloxy) -ethyl ester, 1-diastereomer p-toluenesulfonate

A suspension of 4.88 g (20 mmol) of 7-amino-3-methoxymethyl-3-cephem-4-carboxylic acid in 200 ml of anhydrous methylene chloride is stirred at 0 ° C in 3.12 ml (21 mmol) of DBV. To the slightly cloudy yellow solution was added 4.99 g (24 mmol) of 1-bromoethyl ester of 2,2-dimethylpropionic acid and stirred at room temperature for 3 hours. The reaction solution is poured into a mixture of 600 ml of saturated sodium bicarbonate and 800 ml of methylene chloride. The organic layer was washed with brine, dried over sodium sulfate and evaporated in vacuo. The resulting product (9.3 g) was dissolved in 15 mL of ethyl acetate and mixed with a solution of 1.9 g (10 mmol) of p-toluenesulfonic acid monohydrate in 10 mL of ethyl acetate. The precipitate of product was filtered off, washed with diisopropyl ether and dried in vacuo. Yield: 3.95 (36%); Diastereomer 1 / Diastereomer 2 = 85/15.

Recrystallization of the salt from n-propanol yields pure diastereomer 1, which is further processed according to the procedures described.

An example

7-Amino-3-methoxymethyl-3-cephem-4-carbonic acid 1- (2,2-dimethylpropionyloxy) -ethyl ester, 1 diastereomer tosylate and 2 diastereomer hydrochloride

By the procedure described in Example 5, 4.88 g (20 mmol) of 7-amino-3-methoxymethyl-3-cephem-4-carboxylic acid is obtained in the form of a crude product. The oil thus obtained was dissolved in 20 ml of ethyl acetate and mixed with 0.65 ml (9.2 mmol) of freshly prepared acetyl chloride solution and 1.07 ml (18.4 mmol) of ethanol in 5 ml of ethyl acetate. After cooling in an ice bath, the precipitated hydrochloride precipitate is filtered off, washed with ethyl acetate and dried. Yield: 2.57 g (32%), 1 diastereomer / 2 diastereomer 2 = 22/78.

The filtrate is mixed with 1.75 g (9.2 mmol) of a solution of p-toluenesulfonic acid monohydrate in 8 ml of ethyl acetate, and the precipitate is filtered off. Yield: 1.15 g (16%), 1 diastereomer / 2 diastereomer = 97/3.

The spectral data are consistent with the product of Example 3.

An example

Stage

3-Methoxymethyl-7 - ((naphth-2-yl) methylideneamino) -3-cephem-4-carboxylic acid 1- (2,2-dimethylpropionyloxy) -ethyl ester

According to the procedure described in Example 5, 7.44 g (10 mmol) of 7-amino-3-methoxymethyl-3-cephem-4-carboxylic acid are obtained in the form of a crude ester. The oil thus obtained was dissolved in 30 ml of anhydrous methylene chloride and mixed with a solution of 1.56 g (10 mmol) of naphthalene-2-carbaldehyde in 40 ml of toluene. After 3 hours at room temperature, dilute with 40 ml of toluene and wash 3 times with 30 ml of water. The solution was dried over magnesium sulfate and concentrated to dryness in vacuo.

Stage

Chromatographic separation of diastereomers The crude product of the step is chromatographed on 500 g of silica gel (pH 7.5). For this purpose, commercial silica gel (35-70 µm) is suspended in water and mixed with dilute sodium hydroxide solution to a constant pH of 7.5. The silica gel is filtered off, washed with methanol and dried at 110 ° C under 20 torr. 1.7 g (33%) of diastereomer 1 was eluted first with toluene / ethyl acetate 20/1 followed by 1.65 g (32%) of diastereomer 2. 1 The diastereomer crystallizes from methanol in the form of colorless needles; melt temp. 110 ° C.

Diastereomer ^T H - NMR (CDCl ₃ 270 MHz): δ = 1.22 (singlet, 9H,

C (CH ₃ ) ₃ ), 1.58 (doublet, 3H, CH-CH ₃ ), 3.22 (singlet,

3H, OCH ₃ ), 3.57 (singlet, 2H, SCH ₂ ), 4.31 (AB-system, 2H, CH ₂ O), 5.21 (doublet, 1H, H-6), 5.50 ( double doublet, 1H, H-7), 6.99 (kvadrupletas, 1H, CH-CH ₃₎

7.52 (mc, 2H, aromatic H), 7.88 (mc, 3H, aromatic H), 8.03 (mc, 3H, aromatic H), 8.78 (doublet, 1H,

CH = N).

Diastereomer ^Τ Η - NMR (CDCl ₃ 270 MHz): δ = 1.22 (singlet, 9H,

C (CH ₃ ) ₃ ), 1.58 (doublet, 3H, CH-CH ₃ ), 3.32 (sunglet,

3H, OCH ₃ ), 3.52 (singlet, 2H, SCH ₂ ), 4.26 (AB-system, 2H, CH ₂ O), 5.26 (doublet, 1H, H-6), 5.49 ( double doublet, 1H, H-7), 7.02 (quadruple, 1H, CH-CH ₃ ),

7.51 (mc, 2H, aromatic H), 7.84 (mc, 3H, aromatic H), 8.02 (mc, 3H, aromatic H), 8.75 (doublet, 1H,

CH = N).

The cleavage of Schiff bases to the pure diastereomers of 7-amino-3-methoxymethyl-3-cephem-4-carboxylic acid 1- / 2,2-dimethylpropionyloxy / ethyl ester is carried out using Girard's T-reagent according to procedures described in the literature (e.g. Kamachi et al. The Journal of Antibiotics XI, I / II (1988), 1602-1616).

DEFINITION OF INVENTION

Claims (7)

DEFINITION OF INVENTION 1. Formula II crystalline addition salts of both diastereomeric acids of 3-cephem-4-carboxylic acid 1- / 2,2-dimethylpropionyloxy / ethyl ester: H ₂ N N8 _c — CONHNOH // Λ ^ χ ^ ³ CO ₂ CHOCC (CH ₃ ) ₃ * HX ^H 3 ^C wherein X is a physiologically acceptable anion of a single base or polybasic inorganic or organic acid and the group = N-OH is in the syn position. 2. The crystalline addition salt of both diastereomeric acids of 1- (2,2-dimethylpropionyloxy) -ethyl ester of 3-cephem-4-carboxylic acid of the general formula II according to claim 1, characterized in that the (C) -labeled C atom has the (IS) configuration. 3. The crystalline addition salt of both diastereoisomers of the 1- / 2,2-dimethylpropionyloxy / ethyl ester of 3-cephem-4-carboxylic acid ester of both diastereoisomers of the general formula II according to claim 1 or 2, wherein HX is benzenesulfonic acid, p-toluenesulfonic acid or 4-ethylbenzenesulfonic acid. 4. A process for the preparation of diastereomeric compounds of the general formula II according to claims 1-3, characterized in that a) obtains a compound of formula III CO ₂ CHOCC (CH ₃ ) separates diastereomers chromatographically, separates trityl groups, obtains acid addition products, or (b) obtains by crystallization from a mixture of diastereomers of formula II a greater quantity of a less polar diastereomer; or c) obtains the intermediate of formula IV in the form of the separated diastereomers: CO ₂ CHOCC (CH ₃ ) ₃ [IV] ^H 3 ^C ° and converts to the separated diastereomers of formula II. Pharmaceutical compositions effective against bacterial infections, characterized in that they contain as active ingredients the crystalline addition salts of both diastereomeric acids of the 3cephem-4-carboxylic acid 1- / 2,2-dimethylpropionyloxy / ethyl ester of the general formula II according to claims 1-3 and conventional carriers and diluents. 6. A process for the preparation of a pharmaceutical composition effective against bacterial infections according to claim 5, characterized in that the 1-2,2-dimethylpropionyloxy / ethyl ester of the 3-cephem-4-carboxylic acid of general formula II is combined with the crystalline acid addition salt of both diastereomers. Converts with conventional carriers or diluents into a suitable dosage form. The crystalline addition salts of both diastereomeric acids of formula II according to claims 1-3 for use in combating bacterial infections.