CS204989B2 - Method of producing cephalosporin antibiotics - Google Patents

Description

CZECHOSLOVAK SOCIALISTIC

REPUBLIC

(19J

DESCRIPTION OF THE INVENTION

PATENT 204989 (Π) (B2)

(22) Registered 20 08 74 (21) (PV 5916-78) (32) (31) (33) Priority from 21 08 73 (39645/73) United Kingdom (40) Published 31 07 30 (51) Int. Cl.3C 07 D 501 / 60C 07 D 501/04 // A 81 K 31/545

OFFICE FOR INVITATION AND DISCOVERY (45) Published 15 12 83 (72) COOX MARIIN CHRISTOPHER, LIVERPOOL, GREGORY GORDON IAN, CHALFONT ST. PETER and BRADSHAW JANICE, HARROW (Big

Britain) (73)

Patent holder GLAXO OPERATIONS UK LIMITED, GREENFORD (United Kingdom) (54) Method of producing cephalosporin antibiotics 1

The present invention relates to a process for the production of antibiotic cephalosporins.

These compounds are named by nomenclature using the abbreviation "cefam" according to J. Am. Chem. Soc., 1962, 84, 3400, wherein the term "refinement" refers to a basic single-bond cefam structure.

A number of cephalosporin compounds with varying degrees of antibacterial activity are known, the compounds having a double bond at the 3-position and usually at the 3-position substituted with a methyl group or a substituted methyl group and at the 7-position acylamide group. It is well known that the antibiotic properties of certain 3-cefem-4-carboxylic acids are predominantly determined by the nature of the substituents in position 7β and in position 3. Attempts have been made to find suitable combinations of these groups to give compounds with good antibiotic properties .

Cephalosporin antibiotics are widely used in humans and animals, for example in the treatment of diseases caused by bacteria resistant to other antibiotics, for example penicillin compounds, and to the treatment of penicillin-sensitive, cephalosporin, effective pro-gram-positive and Gram-negative microorganisms, while cephalosporin antibiotics with a broader spectrum are still needed.

The practical use of a number of cephalosporin antibiotics is limited by the relatively high sensitivity to β-lactamases of various bacteria. Another disadvantage of cephalosporin antibiotics is that they undergo in vivo degradation. Such antibiotics of this type are deactivated in this manner, often very rapidly, predominantly by the action of enzymes, for example the esters of the human or animal organism.

Another difficulty with many cephalosporins for therapeutic use is that they undergo degradation in vivo. Thus, a significant number of known cephalosporin antibiotics have been found to be deficient in their inactivation by enzymes (e.g., esterases) after administration, often rapidly.

It has been found that a particular class of antibiotic with a particular combination of 7β-acylamidocapane and a substituent at the 3-position has a broad spectrum of activity as well as a high stability to (3-lactamases and good stability in the organism. the amide group of 2-aryl-2- (etherifiedoxyimino) acetamido group, which is substantially in the form of a syn-isomer, wherein the 3-position is a carbamoyl-oxymethyl group 204989 204989 4

N «NCO

Accordingly, the invention is a process for the preparation of anti-biotics of formula I (III) OR *

wherein R 1 is furyl, thienyl or phenyl, R 2 is alkyl of 1 to 4 carbon atoms, cycloalkyl of 3 to 7 carbon atoms or phenyl,

R a is hydrogen or an ester-red group providing physiologically harmless esters, in the form of a syn-isomer or in a mixture of a syn-isomer and an anti-isomer containing at least 90 o / o syn-isomer as well as their physiologically harmless salts when hydrogen is hydrogen or 1-oxides.

The most preferred compounds of the invention are the syn-isomers of Formula I, substantially free of the corresponding anti-isomers.

According to the invention, the process is carried out at a temperature of -50 to +50 ° C until the compound of formula II is reacted.

where

Acyl means a group R1. what. wherein R 1 and R 2 are as defined above, B is S or S - O, R 11 is hydrogen or a carboxyl blocking group such as diphenylmethyl or tert-butyl, the dashed line between positions 2, 3 and 4 molecules indicate that the compound may be a 2- or 3-cephem compound, with a carbamoylating agent that is iso-

a cyanate of the formula III wherein R13 denotes a labile group, such as mono-, di- or trichloroacetyl, or chlorosulfonyl, the A2-isomer is converted to the desired AMsomer and / or the carboxyl- or N-protecting group is removed and / or reducing the cephalosporin sulfoxide to the corresponding sulfide, and subsequently isolating the compound of formula I optionally after separation of the syn-isomer and the anti-isomer and optionally converting the compound of formula I to its 1-oxide (if desired) where R a represents hydrogen) or physiologically acceptable salts or esters thereof.

The process according to the invention is preferably carried out by isolating the compound of the formula I or a physiologically acceptable salt thereof (when R a is hydrogen) or its 1-oxide in the form of a syn-isomer free of the anti-isomer. To produce preferred compounds of Formula I wherein R 1 is 2-furyl, the compound of Formula II wherein R 1 is 2-furyl is used as the starting material. Similarly, in the preparation of other preferred compounds of formula (I) wherein R2 is ethyl, tert-butyl, cyclopentyl or phenyl, the compounds of formula (II) wherein R2 is as defined above are used as starting materials.

The compounds of the invention are found to be the syn (cis) isomeric form due to the configuration of the group -OR 2 to the carboxamidosine. In the description of the present invention, the syn configuration is preferably as follows: R1. C. CONH—

N OR2.

The syn configuration is marked this way, according to Ahmad and Spenserv Can. J. Chem. 1961, 39, 1340.

The salts of the compounds of formula (I) may be salts with inorganic bases, for example, alkali metals such as sodium and potassium, alkaline earth metals, talc or salts with organic bases such as procaine, phenylethylbenzylamine, dibenzylethylenediamine, ethanolami- diethanolamine, triethanolamine and N-methylglucosamine. The salts may also be in the form of resinates, for example with polystyrene resin or copolymer of polystyrene and divinylbenzene with crosslinks and containing amino or quaternary amino groups. In the case where R 1 in formula I is furyl, it may be 2-furyl or 3-furyl, if it is thienyl, it may be 2-thienyl 204989

S or 3-thienyl. A preferred value for R 1 is 2-furyl.

As already mentioned, R2 in formula I is alkyl of 1 to 4 carbon atoms, for example methyl, ethyl or tert-butyl, cycloalkyl of 3 to 7 carbon atoms, for example cyclopentyl or phenyl.

The compounds of the invention possess valuable properties, namely high potency against a large number of Gram-positive and Gram-negative microorganisms, and at the same time high stability against β-lactamases of various gram-negative organisms and good in vivo stability, especially in the case of esterase activity.

The compounds of the invention are preferably used to treat bacteria and animal diseases caused by bacteria. An important compound of the invention is [6R, 7R) -3-carbamoyloxymethyl-7-

- [2- (fluoro-2-yl) -2-methoxyiminoacetamido] -3-cephem-4-carboxylic acid in the form of a syn-isomer represented by the general formula IV

r Η H jL-C. CONH-i-SiO 2 XOCH 3 OCOOH flV) usually in the form of an alkali metal salt, especially in the form of a sodium salt. This compound is active on a wide range of gram-negative and gram-positive microorganisms, for example Staphylococcus aureus, Strepococcus pyogenes and Streptococcus vilidans, Diplococcus pneumoniae, Haemophilus fluenzae, Neisseria and Clostridia species, Escherichia coli, Klebsiella, Proteus and Entebracter species, as demonstrated in vitro in vivo. The compounds are particularly potent in vitro, particularly against Haemophilus influenzae, Neisseria gonorrhoeae and Neisseriameningitidis. The compound is very stable to pro-beta-lactamases of a number of gram-negative organisms, as demonstrated, for example, by invitro against enzymes of various strains of Escherichia, Enterobacter and Klebsiella. The compound is resistant to the action of mammalian esterases and thus stable in both human and animal organisms, as can be demonstrated by the high-level unchanged compound in urine. The high serum levels of these compounds develop after parenteral administration in humans and animals, with little serum binding. The use of highly soluble alkaline salts, for example sodium salts, is advantageous since these salts are very rapidly distributed throughout the body after administration.

Sodium (6R, 7R) -3-carbamoyloxymethyl-7- [2- (fur-2-yl) -2-methoxyiminoacetamido] -3-cefem-4-carboxylate was found to the isomer is present in a variety of crystalline forms, including solvates, all of which are within the scope of the invention.

Typically, the sodium salt is prepared by reacting a sero-solution of a compound of formula IV in a polar solvent, such as dimethylacetamide, in a mixture of these solvents, such as in a mixture of acetone dimethylacetamide or dimethylamide and formamide, or in polar water / organic mixtures. solvents, for example in a water / acetone mixture with a small molar excess of sodium 2-ethylhexanoate, as a solution in a suitable alcohol, such as ethanol, in a ketone, such as acetone, in a chlorinated hydrocarbon such as methylene chloride , in an ester such as ethyl acetate or an ether such as dioxane, usually at room temperature, and the precipitated salt is separated off after optionally cooling the solution, for example to 4 ° C. When substantially free of solvents are used, (6R, -7R) -3-caramoyloxymethyl-7- [2- [f-2-yl] -2-methoxyiminoacetamido] -3 is obtained. Sodium 4-carboxylate syn-isomer. This material contains approximately 1.5% water. When the solvent contains more than 2% water, the result is a reaction form II which also contains about 2% water. When the solvent contains more than 60% dioxane, a salt of Form III is obtained, the material being a dioxane solvate containing about 1 mole of dioxane. Form II can also be obtained by using a solvent with a water content of about 60 to 80 ° C. Form I, II or III may also be obtained by crystallization of the same compound from the lyophilized form in the appropriate anhydrous, water-containing or dioxan-containing solvent. When form I is exposed to water vapor, for example at a relative humidity of 75%, the salt absorbs a further amount of water and its crystalline form changes to form IV, which contains about 4% water, i.e. about 1 mole, and it is therefore probably a hydrate. This form is reversible so that Form IV can be converted back to Form I, for example, by vacuum drying with phosphorus pentoxide. Form II does not absorb additional water, but can be converted to Form I by heating the suspension of Form II in boiling methanol.

Form II can be precipitated in gel form, which can then be dried under vacuum to give a solid having a high density, very little crystalline or amorphous. However, crystalline form III can also be obtained by treating the sodium salt aqueous solution with a large excess (e.g., 8 volumes) of dioxane, optionally in the presence of a small amount of ethanol, and then separating the resulting white needle crystals after cooling to 4 ° C, after washing the product with dioxane 204989, and drying the crystals, for example, at 20 ° C.

Form III is hygroscopic and, when exposed to water vapor, e.g., at 75%, this form of dioxane is lost and converted to form IV, which can then be dried, for example, by phosphorus pentoxide form I. -bem processes crystalline form III, the crystalline form does not change. Form III is four forms of (6R, 7R) -3-carbamoyl-methoxymethyl-7- [2- (fur-2-) -2-methoxyiminoacetamido] cef-3-em-4-carboxylate sodium salt (syn-isomer), as described above, were characterized in X-ray in the form of a powder (d-distance and intensity) and spectra in infrared light:

X-ray values of powder samples:

but convert to Form I by heating in hot methanol, then loss of crystalline Camera: Debye-Scherrer, radius 114.6 mm irradiation: copper Ka = 1.5418 10 ~ 10 mintensity (I) by visual comparison with calibrated standard: I forms in the case of crystalline form Form I d, that form III has been used herein. I d 8.33 80 3.05 4 7.44 4 2.93 14 (wide) 6.85 45 2.72 8 poorly separated 6.38 5 2.69 10 poorly separated 5.86 4 2.57 9 5 , 36 4 2.47 6 4.82 100 2.40 10 4.56 35 2.35 10 4.36 6 2.26 4 4.19 40 2.20 3 3.95 26 2.11 8 (Wide) 3.82 24 2.04 3 3.62 28 1.94 4 3.47 28 1.89 5 3.32 10 1.82 6 (wide) 3.19 10 1.77 2 Form II d I d I 8 , 78 60 3.49 14 7.81 9 3.07 6 (wide) 6.65 25 2.91 8 4.68 100 (wide) 2.77 6 4.45 10 2.32 3 (wide) 4, 20 10 2.19 2 3.76 20 (broad) 2.08 2 Form III d I d I 14.98 60 4.29 20 12.95 40 4.16 100 10.16 20 3.81 25 8.23 45 3.60 20 7.52 5 3.47 5 6.61 65 3.32 10 6.08 3 3.26 30 5.57 20 3.13 17 4.98 40 2.43 10 4.73 60 2 , 15 15 (broad) (broad) 204989

Form IV

IQ

d I

d I 8.85 70 7.80 6 7.15 25 6.01 20 5.06 18 4.65 100 4.30 25 4.0.1 25 IR spectrum Spectrometer - Perkin-Elmer 521, Area Forma I 3520 w +1590 s 1262 m 3460 m 1556 m 1248 m 3370 m 1534 s 1170 m 3265 s 1480 m 1152 m 1770 sh 1410 s 1134 w 1752 s 1400 s 1112 m 1706 s 1338 s 1076 m 1660 s 1328 s 1054 sh 1620 s 1284 m + 1042 s Forma II 3526 m 1544 s 3492 w 1478 m +3364 m 1412 s 3250 m 1393 s 1758 s 1332 s 1695 s 1284 m 1665 s 1268 m 1642 sh 1240 m 1624 s 1172 m Form III 3465 m 1532 s 3415 m 1482 m 3345 m 1412 s 3275 m 1395 s 3200 m 1326 s 1780 s 1285 m 1702 s 1260 m 1660 s 1230 m 1632 m 1225 m 1618 s 1196 w 1552 m 1180 m form IV 3585 w 1594 s 1264 m 3520 w 1555 m 1240 m 3370 m 1540 m 1172 w +3260 s 1478 m 1152 m 1758 s 1410 s 1114 w 1712 s 1400 s 1078 m 1664 s 1330 s 1058 m 1620 s 1285 m 1042 m 3.75 35 3.10 1 2.93 4 2.76 12 2.62 1 2.41 2 2.30 3 4000—650 cm-1 spectra recorded in nu jungle (nujol stripes were excluded) 1004 m 754 m 978 m918 w882 s878 sh838 w814 w790 w778 w 1 152 m 920 m 1142 m 882 m 1112 m 878 m 1080 m 840 m 1058 m 818 m +1045 m 792 w 1005 m 752 m 980 m 954 w 1155 m 820 w +1124 s 800 w 1078 m 790 w +1058 s 768 m 1048 m 748 m 1014 m 985 m 938 m 888 m 878 m 836 w 1008 m 788 w 980 m 752 m 956 w 920 w 882 m 878 w 838 w 818 w 11 12 s = strong sh = hump m = medium w = weak-stripes, characteristic, for a particular crystalline form When the compound (I) is to be present as an insoluble salt for some use, the salt can be prepared in a conventional manner, for example by reaction with an appropriate amine.

In the compound of formula (II) above, R 11 is a hydrogen atom or a carboxyl blocking group, such as an aliphatic or araliphatic alcohol moiety forming an ester, phenol, silanol or stannan, or a symmetric or mixed anhydride moiety derived from acid. The aforementioned physiologically innocuous carboxylic acid derivatives of general formula 1 (in the case where R a is hydrogen) can be produced by any known method. Solise bases can be formed, for example, by reacting cephalosporinic acid with sodium or potassium 2-ethylhexanoate. Biologically acceptable esters can be obtained using conventional esterification agents. 1-oxides can be obtained by treating the corresponding sulfides with an oxidizing agent, for example, a peracid such as methanediol, peracetic acid, monoperphthalic acid, or chloroperbenzoic acid, tert-butyl hypochlorite, for example in the presence of a weak base such as pyridine .

This carbamoylating agent of general formula III serves to form (at the 3-position of the cephalosporin compound) the N-protecting caramoyloxymethyl group of the general formula -CH 2 O.CO.NHR 11 where R 13 is as defined above, and this group can then be converted to the desired unsubstituted 3-carbamoyloxymethyl-cleavage group R13, for example by hydrolysis. The readily cleavable R13 groups are, for example, chlorosulfonyl, bromosulfonyl, arylalkyl groups such as benzyl, p-methoxybenzyl and diphenylmethyl, tert-butyl, halogenated lower alkanoyl groups such as dichloroacetyl atrichloroacetyl and halogenated lower alkoxycarbonyl groups such as 2,2,2-trichloroethoxycarbonyl. Typically, R13 of this type, except for arylalkyl groups such as diphenylmethyl groups, can be removed by acidic or basic hydrolysis, for example by basic hydrolysis using sodium bicarbonate. Halogenated groups such as chlorosulfonyl, trichloroacetyl and 2,2,2-trichloroethoxycarbonyl can also be cleaved by reduction, while chloroacetyl can be cleaved off with urea thloamides. Arylalkyl groups such as diphenylmethyl may be cleaved with an acid, such as a strong organic acid such as trifluoroacetic acid.

Suitably, the carbamoylating agent of formula III is used in excess (e.g. at least 1.1 moles relative to the compound of formula II). The carbamoylation can be facilitated by the presence of a base, for example a tertiary organic base such as a lower trialkylamine (e.g., triethylamine) or by using an acid of formula II in the alkali metal salt form (e.g., sodium), although such facilitation may not be necessary if the more active isocyanates, for example compounds of formula III, wherein R 13 is a strongly electron withdrawing group, such as at for example, chlorosulfonyl or trichloroacetyl. The carbamoylation reaction of the free acids of formula II with an excess of isocyanates of formula III wherein R 13 is chlorosulfonyl or trichloroacetyl are therefore particularly advantageous for their simplicity, since in this case it is not necessary to temporarily block and then unblock the carboxyl group in position 4 of the cephalosporin nucleus and also because it is moiety no vel-I to remove group R13 in Equals NEM-cephalosporins, such as hydrolysis vevodném solution of sodium bicarbonate-ho.

It will be appreciated that it may be desirable to maintain or introduce an N-substituent group R 13 during the conversion to 3-carbamoyloxymethyl compounds, since there is no undesirable side reaction involving carbamoyloxymethyl. The 3-hydroxymethyl starting materials for use in the process of the present invention can be prepared by the method described, for example, in British Patent No. 1,121,308 and U.S. Patent No. 783,449.

The blocking group at the 7-carboxyl group of the compounds of formula (II) is a desirable group which can be easily removed, preferably a group having from 1 to 20 carbon atoms, for example one of the well-known groups for this purpose. Preferred groups include, for example, lower arylalkoxycarbonyl groups such as p-methoxybenzyloxycarbonyl, p-nitrobenzyloxycarbonyl and diphenylmethoxycarbonyl, lower alkoxycarbonyl groups such as tert-butoxycarbonyl and lower haloalkoxycarbonyl groups. groups such as 2,2,2-trichloroethoxycarbonyl. This group can then be removed in a known manner, for example by acidic, basic or enzymatic-catalyzed hydrolysis. When the sulfoxide of the compound of formula I is purified at the end of the reaction, the corresponding sulfide can be reacted, for example, by reducing the corresponding acyloxysulfonium salt or alkyloxysulfonate salt prepared directly in the reaction mixture by reaction with, for example, acetyl chloride in the case of the acetoxysulfonium salt, wherein the tatoreduction is carried out, for example, with dithionide hydrogen or an iodide ion, for example a solution of potassium iodide in a water miscible solvent such as acetic acid, tetrahydrofuran, dioxane, dimethylformamide or dimethylacetamide. The reaction is carried out at -20 to + 50 ° C. When the reaction product is a 2-cephem-4-carboxylic acid ester, the desired 2-cephem compound can be obtained by treatment with a base.

The antibiotic compounds of the present invention can be formulated into pharmaceutical formulations for suitable routes of administration in a manner similar to other antibiotics; in particular, compounds of the formula I or their physiologically acceptable salts (in the case of Ra in the general formula I, hydrogen) can be processed from the compounds according to the invention or modified for use in human or veterinary medicine. They are used together with conventional pharmaceutical carriers or excipients for administration.

The antibiotic of the invention may be a pro-injection in ampoules or in a larger amount with a preservative. They may be suspensions, solutions or emulsions in oil or aqueous medium, optionally using suspending, stabilizing and / or dispersing agents. The active ingredient may also be in the form of a powder which dissolves just prior to administration, for example in sterile, pyrogen-free water.

For example, in veterinary medicine, there may be intramammary preparations (for administration to the mammary gland) either in long-acting non-fast-acting vehicles. The formulations usually contain at least 0.1 percent, for example 0.1 to 99 percent, preferably 10 to 60 percent, of the active ingredient, depending on the route of administration. The single dose preferably contains 50 to 1500 mg of active ingredient. Adult humans are preferably administered from 500 to 4000 mg per day, depending on the frequency of administration.

The compounds of the invention may also be administered in combination with other compatible drugs, such as antibiotics, for example penicillins, other cephalosporins or tetracyclines.

The invention will be illustrated by the following examples. All temperatures are in degrees Celsius, melting points were determined on a Kofer block. Example 1

Syn-isomer of (6R, 7R) -3-carbamoyloxymethyl-7- [2- (fur-2-yl) -2-methoxyiminoacetamido] -3-cephem-4-carboxylic acid

Method (i) 750 ml of acetone was cooled to 0 ° C and treated with 28.8 ml of trichloroacetyl isocyanate (250 mmol) and the resulting solution was re-cooled. 45.6 g (120 mmol) of syn-isomer of (6R, 7R) -7- (2- (fur-2-yl) -2-methoxyiminoacetamido] -3-hydroxymethyl-3-cephem are added -4-carboxylic acid to the stirred i-cyanate solution in portions at 5 minute intervals so that the reaction temperature does not exceed 6 DEG C. The yellow solution is stirred for a further 15 minutes and 4.5 ml of methanol are added. The solution is concentrated to 60 ml and the concentrated solution is dissolved in 750 ml of methanol and sodium bicarbonate (45.3 g, 540 mmol) in water (600 ml) is added, followed by the addition of activated charcoal (4.5 g) and The suspension is stirred at room temperature for 2 hours and the activated carbon is removed by filtration through diatomaceous earth, the pH of the light yellow filtrate is adjusted to pH 4.5 with dilute hydrochloric acid. The pH is adjusted to 2.0 with dilute hydrochloric acid and the product is isolated by filtration, washed with water (3X1). 50 ml) and dried at 40 ° C under vacuum for 16 hours to give the title compound (37.46 g, 74.5%), [α] D 20 + 63.7 ° (c 1.0, 0.2 M). pH 7 phosphate buffer), Amax (pH 6 phosphate buffer) 274 nm (ε17600). Infrared and NMR-spectroscopy data confirm the structure of the resulting compound.

Method (ii)

The acid formed by the syn-isomer of (6R, 7R) -7- (2- (fur-2-yl) -2-methoxyiminoacetamido) -3-hydroxymethyl] -3-cephem-4-carboxylic acid (3.81) g, 9.55 mmol) in a mixture of 70 mL of dichloromethane and 25 mL of tetrahydrofuran was treated with dichloroacetyl isocyanate (2.6 mL, 25 mmol) at 5 ° C. The reaction mixture is then treated in the same manner as in method (i) to give the final compound (3.36 g, 83.0%), [α] η 2θ + 63 °, Arnux 273.5 nm (ε 17). 800) with a similar infrared and NMR spectrum as the product of method (i).

Method (iii)

The acid formed by the (6R, 7R) -7- [2- (f-2-yl) -2-methoxyiminoacetamido] -3-hydroxymethyl-3-cephem-4-carboxylic acid syn-isomer ( 19.05 g (50 mmol) in anhydrous acetonitrile (250 mL) was treated with chlorosulfonyl isocyanate (6.33 g, 75 mmol) in acetonitrile (80 mL) at a temperature between 5-10 ° C.

The reaction mixture is stirred at 0-5 ° C for 10 minutes and 50 ml of water are added.

The mixture was stirred at about 20 ° C and after 20 minutes a white crystalline solid was separated. 204989 15 16

Evaporation and filtration provide the final compound, (18.17 g, 85.7%, [α] D 20 + 62.5 °, Amax 273.5 nm (ε 17,820), with an infrared and NMR-spectrum similar to that obtained by the method (i) A second yield (1.88 g, 8.6%) of the product with similar constants was obtained by evaporation of the mother liquor.

Form I syn-isomer of (6R, 7R) -3-carbamoyloxymethyl-7- [2- (fur-2-yl) -2-methoxyiminoacetamido] -3-cephem-4-carboxylate sodium Method (i)

Per 100.0 g of syn-isomer of (6R, 7R) -3-carbamoyloxymethyl-7- [2- (fur-2-yl) -2-methoxyiminoacetamido] -3-cephem-4-carboxylic acid in the mixture N, N-dimethylacetamide (400 mL) with 1 L of acetone was treated with 40 g of sodium 2-ethylhexanoate in 200 mL of acetone. The mixture was loaded and shaken at ambient temperature for 1.25 hours. The product was filtered off, washed with acetone (500 mL) and then precipitated with acetone (3X 300 mL) and finally precipitated with ether to give the title compound (101.4 g, 92.5%) containing 0 equiv. 65 molekvivalentůvody. The product has a [w] D 2 O + 61 ° (c 0.5, pH 4.5 phosphate buffer) and an A max of 273 nm 1% E 1 z 2 → 412 (H 2 O). Infrared and NMR-spectroscopy data confirm the structure of the resulting compound and the infrared spectrum indicates that the compound is a salt of Form I.

Method (11)

Method (i) is repeated except that the cephalosporinic acid is initially dissolved in a mixture of N, N'-dimethylacetamide with ethanol denatured with methanol instead of a mixture of N, N-dimethylacetamide and acetone. the resulting compound (80%) having properties similar to the product obtained by the method of (i). The infrared spectrum indicates that the compound is a salt of Form I.

Method (iii) 4.24 g (10 mmol) of syn-isomer of (6R, 7R) -3-carbamoyloxymethyl-7- (2- (fur-2-yl) -2-methoxyiminoacetamido] -3-cephem- 4-carboxylic acid was dissolved in N, N-dimethylacetamide (20 mL), which was dried with a molecular sieve (Linde 4A) for 24 hours. A solution of 2-ethylhexanoate phosphoric acid in ethyl acetate (80 mL) was added thereto. (2.0 g, 12 mmol, recrystallized from dioxane and dried over oxygen, dehydrated with molecular sieve (Lin 4A) for 24 hours) The solution is stirred in a closed vessel for about 15 minutes until crystallization begins and then The product was filtered off, washed with anhydrous ethyl acetate (about 100 mL) and, if still wet, transferred to an oven and dried at 20 ° C under vacuum over phosphorus pentoxide overnight. to give the title compound (3.89 g, 87%).

The IR and NMR spectra of the product agree with those of an authentic sample

Sodium Form (2R, 7R) -3-Carbamoyloxymethyl-7- [2- (fur-2-yl) -2-methoxyiminoacetamido] -3-cephem-4-carboxylate-4-carboxylate Form (i) 0, 2 g of activated carbon is added to a solution of the (6R, 7R) -3-carbamoyl-oxymethyl-7- [2- (fur-2-yl) -2-methoxy-iminoacetamido] -3-cephem-4- carboxylic acid (4.00 g, 9.42 mmol) in a mixture of 132 mL of acetone and 1.33 mL of water. The suspension is stirred for 30 minutes and filtered through a bed of diatomaceous earth, washing the filter bed with 10 ml of acetone. A filtered solution of sodium 2-ethylhexanoate (1.66 g, 10 mmol) in acetone (20.0 mL) was added to the stirred filtrate for one hour. The resulting slurry was stirred for an additional 10 minutes and the white precipitate was filtered off, washed with acetone (2 x 25 mL) and dried in vacuo to give the title compound (4.06 g, 93.0%). 0.91, H2O), Amax (H2O) 274nm (ε 17,400), [found: C: 41.0, 41.2, H: 3.45, 3.6, N: 12.3, 12.4] Na: 5.2, S: 6.6, 6.85, H 2 O: 2.7, 2.7%.

For CieHtsNéNaOeS. 0.7 H2O (459.0) calculated: C: 41.8, H: 3.6, N: 12.2, Na: 5.0, S: 7.0, H2O: 2.7%], purity by HPLC 99.4%. The NMR spectrum of the resulting compound was an authentic sample and the infrared spectrum indicated that the product was form II salt of the antibiotic.

Method (ii)

Syn-isomer of (6R, 7R) -3-carbamoyl-oxymethyl-7- [2- (f-2-yl) -2-methoxy-iminoacetamido] -3-cephem-4-carboxylic acid (16.98 g (40 mmol) was added to the stirred mixture of 333 mL of acetone and 8.5 mL of water. Treatment with charcoal and filtration of this solution slowly added sodium 2-ethylhexanoate (7.32 g, 44 mmol) in acetone (85 mL) over an hour. The reaction mixture was stirred for 15 18 minutes, filtered and the product washed with acetone (2 x 65 mL) and dried under vacuum at 20 ° C overnight to give the title compound (17.95 g, 93.5). %) containing 0.5 molFkO. The NMR spectrum of the product is confirmed to correspond to that of an authentic sample, the infrared spectrum indicates that the product is the salt of Form II. Attaches

Sodium Form III isomer of (6R, 7R) -3-carbamoyloxymethyl-7- [2- (ur-2-yl) -2-methoxyiminoacetamido] -3-cephem-4-carboxylate 4.0 g syn Dissolve (6R, 7R) -3-carbamoyl-oxymethyl-7- [2- (fur-2-yl) -2-methoxy-iminoacetamido] -3-cephem-4-carboxylate sodium in 20 ml of water . 20 ml of methanol and 160 ml of dioxane are added and the solution is filtered and then left to crystallize at 4 ° C. The very white, needle-like crystals are filtered off, washed with 100 ml of dioxane and, if still wet, transferred to an oven and dried at 20 DEG C. under vacuum to give the title compound (3.96 g, 78.5%). ). The infrared and NMR spectra of the product correspond to that of an authentic sample. Example 5

Form IV syn-isomer of (6R, 7R) -3- -carbamoyloxymethyl-7- [2- (fur-2-yl) -2-methoxyiminoacetamido] -3-cephem-4-carboxylate-4-carboxylate

Samples of Form I and Form III of the syn-isomer of 3-carbamoyloxymethyl-7- [2- (fur-2-yl) -2-methoxyiminoacetamido] -3-cephem-4-carboxylate sodium prepared according to method (iii) Examples 2 and 4 are allowed to wet (75% relative humidity) to give the title compound. The infrared and NMR spectra of the products matched with the autosample. KarlFischer water determination gives 4.0 and 3.85%, respectively (1 mole% is equivalent to 3.9%). Example 6 a) Syn-isomer tert-butyl (6R, 7R) -3-carbamoyloxymethyl-7- [2- (f-2-yl) -2-methoxyiminoacetamido] -3-cephem-4- carboxylate

A suspension of 4.4 g of syn-isomer of (6R, 7R) -3-carbamoyloxymethyl-7- [2- (fur-2-yl) -2-methoxyimino acetamido] -3-cephem-4-carboxylic acid in 200 ml anhydrous methylene chloride is treated with 6.6 ml of 10-tert-butyl-N, N'-dicyclohexylisourea to form a pale yellow solution. After 24 hours at 23 ° C, the starting material remains and 3.3 ml of isourea is added. After 48 hours, the mixture was filtered and the filtrate evaporated under reduced pressure. The resulting material was stirred with a mixture of ether and ethyl acetate to remove the remaining dicyclohexylurea. The filtrate was washed with saturated aqueous sodium bicarbonate solution and water, then dried and evaporated to give a precipitate (5.2 g). Chromatography on silica gel with toluene: ethyl acetate (2: 1) as solvent gave the title compound (3.9 g) as a pale yellow foam, Amax (ethanol) 275.5 nm (ε 400 400). b) Syn-isomer tert-butyl (1R, 6R, 7R) -3-carbamoyloxymethyl-7- [2- (f-2-yl) -2-methoxyiminoacetamido] -3-cephem-4- carboxylate-1-oxide 0.3 ml of tert-butyl hypochlorite is added to a vigorously stirred solution of 0.98 g of tert-butyl (6R, 7R) -3-carbamoyloxymethyl-7 - [2- (f-2-yl)] -2-methoxyiminoacetamido-3-cephem-4-carboxylate in 25 ml of pyridine and 1 ml of water at -45 ° C. After stirring for 2 minutes, 1 ml of 2 N sulfuric acid is added and the mixture is immediately poured into 100 ml of 20% (w / v) orthophosphoric acid. The solution was extracted with ethyl acetate (2 x 100 mL) and the combined organic extracts were washed with sodium bicarbonate solution (100 mL) and water (100 mL) and dried with magnesium sulfate and concentrated in vacuo. The crude product is chromatographed on silica gel plates using ethyl acetate as the eluate, the smaller S-oxidized R-oxide component. Extraction of the slower component with ethyl acetate gave the title compound (0.27 g, 27%). Infrared and NMR spectroscopy data confirm the structure of the resulting compound. c) Syn-isomer of (1R, 6R, 7R) -3-carbamoyloxymethyl-7- [2- (ur-2-yl) -2-methoxyiminoacetamido] -3-cephem-4- 1-oxide carboxylic acid 0.42 g of syn-isomer tert-butyl- (1R, 6R, 7R- 3-carbamoyloxymethyl-7- [2- (f-2-yl) -2-methoxyiminoacetamido] -3-cephem-4 The carboxylate 1-oxide is dissolved in 5 ml of trifluoroacetic acid and stirred at room temperature for 8 minutes, the solution is evaporated to a red oil, dissolved in 5 ml of ethyl acetate-acetone (1: 1). and adding dropwise to 50 ml of petroleum ether (50 to 80 ° C) with stirring. The settled solids or plugs retained by the aluminum sealing rings to prevent the accumulation and drying in a desiccator. the phase was separated and added dropwise to 50 ml of petroleum ether (60 to 80 ° C) to give the title compound as a colorless solid (150 mg, 40%), Amax (0.25 N NaHCO 3) 263.5 ( with 15,000) and 281 nm (ε 13,700), vmax (n 1799 and 1716 (COOH and OCONH 2), 1684 and 1538 (CONH) and 1060 and 1050 cm -1 (S-O), τ (MezSO-de) 204989 19 values include 0.02 (d, J 8 Hz, CONH), 4.17 (dd, 4 and 8 Hz, 7-H), 4.99 (d, J 4 Hz, 6-H) and 6.09 (s, N-OCH 3). Annexes 7 and 7

Syn-isomer of (1S, 6R, 7R) -3-carbamoyloxymethyl-7- [2- (fur-2-yl) -2-methoxyiminoacetamido] -3-cephem-4-carboxylic acid 1-oxide K Sodium (6R, 7R) -3-Carhamoyloxymethyl-7- [2- (fur-2-yl) -2-methoxyiminoacetamido] -3-cephem-4-carboxylate (2.59 g) stirred solution; (25 ml) was added 1.93 g of metaperiodine. The solution is stirred for 30 minutes at ambient temperature and then acidified with 2 N hydrochloric acid added dropwise. The resulting precipitate is collected, washed successively with water, ethanol and ether and dried in vacuo to give the title compound as a white powder (1). , 63 g) [ajn 20 + 113 ° (c 0.86, MezSO), Amax (pH 6 buffer) 264.5 (ε17,200) and 279 nm (ε 15,600), µmax (nujol) 1770 (β-lactam) , 1740 and 1716 (CO 2 H), 1688.1654, 1589 and 1530 (CONH and OCONH 2) and 1039cm-1 (S -> O), τ (MSQ-de) 0.60 (d, J 8Hz, NH), 2, 11, 3.19, 3.31 (multiplet, furylprotones), 4.08 (q, J5 and 8Hz, C-7H), 4.87 and 5.45 (ABq, J13Hz, CH2OCONH2), 4.96 (d, J 5Hz, C-6H), 6.08 (s, OCH 3), 6.10 and 6.42 (ABq, J 18 H 2, C-2 CH 2).

Syn-isomer of (1R, 6R, 7R) - and (1S, 6R, 7R) -3-carbamoyloxymethyl-7- [2- (fur-2-yl) -2-methoxyiminoacetamido] -3-cephem-4 -carboxylic acid

A solution of 1.93 g of sodium periodate in 10 ml of water is added to a stirred solution of (6R, 7R) -3-carbamoyloxymethyl-7- [2- (trifluoromethyl) -2-methoxyiminoacetamido] -3-cephem. -4-carboxylate (2.59 g) in water (25 mL). Stirring is continued for 30 minutes at room temperature and the solution is then acidified with 2 N hydrochloric acid (2 ml). The settled 1S-oxide was filtered off, washed with ethanol (5 mL) and ether (20 mL) and dried in a desiccator to give 1.59 g of a colorless solid, [* x] D + 110 ° (c1, MezSO), corresponding to the product described in Example 7.

The mother liquors are saturated with sodium chloride and filtered. The filtrate was extracted with ethyl acetate (2X 100 mL). Combine the organic extracts, dry with magnesium sulfate and concentrate in vacuo to give a yellow solid. The crude product is washed with acetone and the insoluble material is removed by filtration. The filtrate was evaporated to dryness and the acetone was washed to give 380 mg of 1R-oxide, [α] D -88 ° (c 1, Me 2 SO), τ (Me 2 SO-d 6) with similar data as in Example 6.

Examples of Production of Pharmaceutical Formulations Example A

Dry powder for injection

The sterile syn-isomer of (6R, 7R) -3-carbamoyl-oxymethyl-7- [2- (fur-2-yl) -2-methoxy-iminoacetamido] -3-cephem-4-carboxylate sodium is filled into glass ampoules, labeled with 500 mg or 1.0 g of cephalosporin compound. The filling is carried out under a nitrogen blanket. The follicles are closed using rubber discs of non-gases or microorganisms. The product is intended to be reconstituted with water for injection or other suitable sterile vehicle just prior to administration.

Example B

Intramammary injection (for cattle)

Percentage composition (w / w ° / o)

Sodium (6R, 7R) -3-carbamoyloxymethyl-7- [2- (fur-2-yl) -2-methoxyiminoacetamido] -3-cef-4-carboxylate syn-isomer 8,33 vehicle to 100 , 00 vehicle (composition):

Tween 60 3.00 white beeswax 6.00 peanut oil 91.00

The last three components were heated to 150 ° C for one hour and then cooled to room temperature with stirring. A sterile antibiotic, finely powdered, is added aseptically to this vehicle and the product gently spread on a high speed mixer. The preparation is filled aseptically into sterile deformable aluminum tubes with a filling weight of 3.0 g; each tube contains 250 mg of cephalosporin compound. The product is intended for use in the milk mammary gland.

Claims (5)

204989 21 22 OBJECT OF THE INVENTION A process for the preparation of a cephalosporin anti-biotics of formula (I) wherein R 1 is furyl, thienyl or R 2 is alkyl of 1 to 4 carbon atoms. cycloalkyl having from 3 to 7 carbon atoms or phenyl and R a is hydrogen or an ester-amino group providing physiologically harmless esters, in the form of a syn-isomer or a mixture of syn-isomer and anti-isomer containing at least 90% of syn-isomer, and physiologically non-harmful salts in the case where R a is hydrogen or a 1-oxide, characterized in that at a temperature of -50 to + 50 ° C it brings a compound of formula (II) wherein Acyl is R1C.CO- N2 OR2 wherein R1 and R2 are as defined above, B is sulfur or S-O, R11 is hydrogen or a carboxyl-blocking group such as diphenylmethyl or tert-butyl, the dashed line between the 2, 3, and 4 positions of the molecule indicates that the compound may be a 2-cephem or 3-cephem compound, with a carbamoylating agent that is an isocyanate of formula III R 13 NCO, (III) wherein R 13 is a labile group such as for example, mono-, di- or trichloroacetyl or chlorosulfonyl, the conversion of the A2-isomer to the desired A3-isomer and / or the carboxyl- or N-protecting-blocking group is removed and / or the sulfoxide is reduced cefalosporin to the corresponding sulfide, and then isolating the compound of formula (I), optionally after separation of the syn-isomer and the anti-isomer, and optionally converting the 1-oxide when converting the compound, when Ra denotes hydrogen; or physiologically n harmful soline esters. 2. A process according to claim 1, wherein the compound of formula (I) or a physiologically acceptable salt thereof, in the case of Ra is hydrogen or its 1-oxide is isolated in the form of a isomer-free isomer. 3. A process according to claim 1 or 2 for the preparation of compounds of formula I wherein R1 is 2-furyl, wherein the compound of formula II wherein R1 is 2-furyl is used as the starting material and the other general symbols are as defined above. meaning. 4. A process according to claim 1 or 2 for the preparation of compounds of formula (I) wherein R2 is ethyl, ethyl, tert-butyl, cyclopentyl or phenyl, characterized in that the compound of formula (II) is used as starting material, wherein R < 2 > and the other general symbols are as defined above. 5. A process according to any one of claims 1 to 4, wherein R 1 is 2-furyl, R 2 is methyl, and R a is hydrogen, characterized in that the compound of formula (I) is used as the starting compound. II, wherein R 1 is 2-furyl R 2 is methyl and the other general symbols are as defined above. Severografia, n. P., Plant 7, Most