FI65623C - EXAMINATION OF ANTIBIOTICS ACTIVE (6R 7R) -3-CARBAMOYLOXIMETHYL-7- (2- (FUR-2-YL) -2-METHOXIMINOACETAMIDO) -CEF-3-EM-4-CARBOXYL SYRICATE - Google Patents

Description

r_, „„ AD PUBLICATION, r £ o? W <11) utläggimingsskrift 65 62 o C (45) F. '' t? R. en n.ywΓ-r, ri * 11 -6 1. J4 ^ (51) K ».lkp / lnt.CI.3 c 07 D 501/34 FINLAND —FINLAND (21) PK« ittlh * k * niu · - Ρ «ηί · * η» βΚηΙο | 2456/71 »(22) HakamlipUvt — AmMcnlfipdag 20.08.74 (Fl) (23) Alkupllvt — GUtlghMtdag 20.08. 74 (41) Become a lulklMktl - Bltvtt ofTwKtlf 22.02.75

Patent and register controlled / 44) Nlhtivlkslpanon J »heard | ulkal * un pvm. -

Patent · och registerstyrelsen AnaBkan utlagd och utUkrtfun publlcurad 29.02.84 (32) (33) (31) Requested «tuolkuu * —Bujäril prior! ** 21 .08.73

England-England (GB) 39645/73 (71) Glaxo Operations UK Limited, Greenford, Middlesex, England-England (GB) (72) Martin Christopher Cook, Liverpool, Lancashire, Gordon lan Gregory, Chalfont St. Peter, Buckinghamshire, Janice Bradshaw, Harrow, Middlesex, Engl ant i-England (GB) (74) Oy Kolster Ab (54) Method for antibiotic active (6R, 7R) -3-carbamoyloxymethyl-7- [2- (fur-2-yl) For the preparation of 2-methoxyiminoacetamido [7-cef-3-em-4-carboxylic acid and its non-toxic salts - For the preparation of antibiotic active compounds (6R, 7R) -3-carbamoyloxymethyl-7- [2- ( The present invention relates to a process for the treatment of the antibiotic-active (6R, 7R) -3-carbamoyloxymethyl-7- (1-yl) -2-methoxyiminoacetamido] -ceph-3-em-4-carboxylate. For the preparation of 2- (fur-2-yl) -2-methoxyiminoacetamide-7-ceph-3-em-4-carboxylic acid of formula

H H

^ ^ -C-CO-NH ---- 0 J-N. ^ - CHoO-CO-NHo (I)

"n.« T

0Cl13 COOH

2,65623 which compound is in the form of a syn isomer or a mixture of syn and anti isomers containing at least 90% of the syn isomer, and for the preparation of non-toxic salts thereof.

The process according to the invention is characterized in that a) the general formula? f a h2n— | - [· "> n - CH2O-CO-NHR12 (II) ioOR11........

a compound of formula 12 wherein B is hydrogen or a carboxyl protecting group; and.

R is hydrogen or an N-protecting group, or an acid addition salt thereof, of the formula OC-C00H (111)

OF

with an acylating agent corresponding to an acid of och3, or h) administering a compound of formula ^ C-CO-WH - S ^ (IV)

° J-N. ^ -CH 0H

Nk (fy * OCH 3 C00R11 in which R1 is as defined above) reacts with a carbamoylating agent such as an isocyanate of formula R13-NC0 (VIII)... 13 in which R is a leaving group to give the 3-position a carbamoyloxymethyl group, the nitrogen atom of which may be protected, after which 3 65623 c) if necessary, removing all carboxyl and N-protecting groups, and finally d) isolating the desired compound of formula I, previously if desired converting the compound to a non-toxic salt.

In this patent application, the names of the cephalosporin compounds are based on the name "kefam" (see J. Amer. Chem. Soc., 1962, 8U, 3,00); "kefem" refers to a kefam base structure with a single double bond.

Ko. Several cephalosponin compounds having a certain antibacterial activity are known in the art, which are -unsaturated and usually have a methyl or substituted methyl group as a substituent at the 3-position and an acylamido group at the 7yS position. It is well known that the antibiotic properties of a given cef-3-em-U-carboxylic acid depend mainly on both the 7γ-acylamino group and the substituent at the 3-position. Substantially extensive studies have been performed to find group combinations that provide antibiotics with special properties.

Cephalosporin antibiotics are widely used in the treatment of diseases caused by pathogenic bacteria in humans and animals, e.g. in the treatment of diseases caused by bacteria resistant to other antibiotics (e.g. penicillin compounds) and in the treatment of penicillin-sensitive patients. Many applications require a cephalosporin antibiotic that is effective against both gram-positive and gram-negative microorganisms, and considerable studies have been conducted to develop very broad-spectrum cephalosporin antibiotics.

The use of many known cephalosporin antibiotics is limited by their relatively high susceptibility to the action of many bacterial β> lactamases. Thus, a desirable property of a broad-spectrum cephalosporin antibiotic is near-complete resistance to β-lactamases and β-lactamases produced by gram-negative microorganisms.

An additional difficulty with many medicinal cephalosporin antibiotics is that they are rapidly degraded in vivo. Thus, many known cephalosporin antibiotics have been found to be inactivated by body enzymes, e.g., esterases.

Thus, a new cephalosporin antibiotic has now been invented with a special combination of a 7/3 acylamide group and a substituent in the 3-position, which gives the compound good broad-spectrum activity, as well as high β-lactamase stability and good in vivo stability. Preferably, the compound is a syn isomer, with almost no corresponding anti-isomer.

The novel compound is defined as the syn (cis) isomer depending on the position of the -OCH 2 group relative to the carboxamide group; the structure of the syn configuration is O-C-CO-NH-

II

N \ OCH3

The Syn configuration is based on Ahmad and Spencer's Can. J. Chem., 1961, 39, 13 ^ 0.

The term "non-toxic" as used for the new salts means physiologically acceptable salts in dosage amounts.

Salts formed from a compound of formula I include inorganic base salts, e.g. alkali metal salts such as sodium and potassium, alkaline earth metal salts such as calcium salts, and organic base salts, e.g. procaine, phenylethylbenzylene! ini-, dibenzylethylenediamine, ethanolamine, diethanolamine, triethanolamine and N-methylglucosamine salts. Salts can also be resinates formed, for example, with a polystyrene resin or a crosslinked polystyrene divinylbenzene copolymer resin containing amino or quaternary amino groups.

The invention also includes solvates, especially hydrates.

As mentioned above, the compound according to the invention has a particularly valuable combination of properties and has a high antibacterial activity, affecting both gram-positive and gram-negative organisms. The activity spectrum is broadened by the particularly high stability of the compound with β-lactamases formed by various gram-negative organisms. The compound is preferably stable in vivo, especially with respect to esterases. These properties make the compound useful in the treatment of various diseases caused by pathogenic bacteria in humans and animals.

The compound according to the invention is active against many gram-positive and gram-negative microorganisms, e.g. the following: Staphylococci 5 65623

Including Staphylococcus aureus, Streptococcus pyogenes and Streptococcus vilidans, Diploeoeeus pneumoniae, Haemophilus influenzae, Neisseria and Clostrida species, Escherichia coli, Klebsiella, Proteus and Enterobacter, as evidenced by both in vitro and in vivo experiments. The compound has good in vitro activity against up to 10 organisms / ml and has particularly good in vitro activity against Haemophilus influenzae, Neisseria gonorrhoeae and Neisseria meningitidis strains. The compound has a very high stability against β-lactamases produced by several gram-negative organisms, which is manifested as in vitro activity against various β-lactamase-producing strains of Escherichia, Enterobacter and Klebsiella species. The compound is resistant to mammalian esterases and thus persistent in the human and animal body, as evidenced by the high yields of the unchanged compound in the urine. It was also found that by parenteral nebulization of the compound in humans and animals, a high blood serum concentration and a low serum binding were achieved.

In medicine, it is preferable to use highly soluble female salts of the compound (e.g., an alkali metal salt such as the sodium salt) because they are rapidly distributed in the body after injection.

Sodium (6R, 7R) -3-carbamoyloxymethyl 7- [2- (fur-2-yl) -2-methoxyiminoacetamide] cef-3-em-U-carboxylate (syn isomer) has been found to be present in various crystalline forms, including solvates, all of which are within the scope of the present invention.

The sodium salt is most conveniently prepared by mixing a solution of a compound of formula I in a polar organic solvent (e.g. dimethylacetamide), a mixture of such solvents (e.g. dimethylacetamide and acetone or a mixture of dimethylformamide and technical, denatured alcohol) or a polar, organic and organic solvent and water. acetone) and a slight molar excess of sodium 2-ethylhexanoate dissolved in a suitable organic solvent (e.g. an alkanol such as ethanol, a ketone such as acetone, or a chlorinated hydrocarbon such as methylene chloride, or an ester such as ethyl acetate, or an ether such as dioxane). most conveniently at room temperature, and isolating the precipitated salt if desired after cooling the solution (e.g. to U ° C).

Using almost anhydrous solvents in this process gives "Form I" sodium (6R, 7R) -3-carbamoyloxymethyl-7-{{(2- (fur-2-yl) -2-methoxyiminoacetamide] cef-3-em-H). carboxylate (syn isomer) containing about 1.5% water. When the solvent system contains more than about 2% water, "Form II" is obtained, which normally contains about 2% water. When the solvent system contains more than n.

60% dioxane is normally obtained as "Form III", the dioxane solvate of which contains 6 65623 about 1 mole of dioxane, although "Form II" can be obtained by using a wet solvent system at an elevated temperature, e.g. 60 ° -80 ° C. Crystallization of amorphous, lyophilized sodium (6R, TR) -3-carbamoyloxymethyl-7- [2-fur-2-yl) -2-methoxyiminoacetamide] cef-3-em-1 * carboxylate (syn-isomer) from suitable dry, aqueous or dioxane-rich solvent systems give the salt of Form I, II or III, respectively.

By placing the "Form I" salt in water vapor (e.g., 75% relative humidity), the salt absorbs more water, and generally converts from its crystalline form to the "Form IV" salt. The material formed contains about 1% by weight of water (i.e. about 1 mole) and is believed to be a hydrate. This change is reversible, so that the "Form IV" salt can be converted to the "Form I" salt, e.g., by drying in vacuo in the presence of phosphorus pentoxide. The "Form II" salt no longer absorbs water upon exposure to water vapor, but can be converted to the "Form I" salt by heating (e.g., about 5 minutes) a suspension of a compound of Formula I in near boiling methanol.

The "Form III" salt obtained by reacting a compound of formula I as described above with sodium 2-ethylhexanoate in dioxane-rich solvent systems is normally precipitated as a gel which can be dried in vacuo to give a solid with a very low bulk density which is little or not crystalline. However, the crystalline salt of "Form III" can be obtained by treating an aqueous solution of the sodium salt with a substantial excess (e.g., about eight times the volume) of dioxane, if desired in the presence of a small amount of methanol, isolating the resulting white, needle-like crystals, preferably by cooling (e.g., 14 ° C), washing. by drying the crystals (e.g. under vacuum at 20 ° C).

The "Form III" salt is hygroscopic and in water vapor (e.g. 75% relative humidity) it loses all dioxane and is converted to a "Form IV" salt which can be dried (e.g. with phosphorus pentoxide) to a "Form I" salt. If the "Form III" salt is treated in this way, the crystallinity of the product appears to be maintained at all stages of the transformation. The "Form III" salt can also be converted to the "Form I" salt by heating a suspension of the "Form III" in near boiling methanol. This change does not cause a loss of crystallinity when using a "Form III" salt.

The four forms of the sodium (6R, 7R) -3-carbamoyloxymethyl-7-Z2- (fur-2-yl) -2-methoxyiminoacetamido-7-ceph-3-em-U-carboxylate (syn-isomer) described above are identified by the following by means of X-ray spectra (d-intervals and intensities) and IR spectra:

Hein tgend 1 f f key meter i a

Camera: Debye-Seherrer, radius ll) i, b mm Radiation: copper Κ ^ = 1.5 ^ 18 Ä

Intensities (I) by comparison with a visually calibrated standard.

Form I

7 65623

AI AI

8.33 80 3.05 k 7.1 + 1 + H 2.93 11+ (broad) 6.85 1 + 5 2.72 8) poorly 6.38 5 2.69 10) separated 5.86 1+ 2.57 9 5.36 k 2.1 + 7 6 1 +, 82 100 2.1 + 0 10 1 +, 56 35 2.35 10 1 +, 36 6 2.26 1+ 1 +, 19 1 + 0 2.20 3 3.95 26 2.11 8 (broad) 3.82 2l + 2, Oi * 3 3.62 28 1.91+! + 3.1 + 7 28 1, 89 5 3.32 10 1.82 6 ( broad) 3.19 10 1.77 2

Form II

AI AI

8.78 60 3.1 + 9 11+ 7.81 9 3.07 6 (broad) 6.65 25 2.91 8 1 +, 68 100 (broad) 2.77 6 1 +, 1 + 5 10 2 .32 3 (broad) 1 +, 20 10 2.19 2 3.76 20 (broad) 2.08 2 8 65623

Form III

d III

1U, 98 60 it, 29 20 12.95 itO U, 16 100 10.16 20 3.81 25 (broad) 8.23 i + 5 3.60 20 7.52 5 3, lt7 5 6.61 65 3 .32 10 6.08 3 3.26 30 5.57 20 3.13 17 (broad) k, 98 kö 2, lt3 10 it, 73 60 2.15 15

Form IV

d lii 8.85 70 3.75 35 7.80 6 3.10 1 7.15 25 2.93 it 6.01 20 2.76 12 5.06 18 2.62 1 it, 65 100 2, lt1 2 it, 30 25 2.30 3 it, 01 25 IR spectra

Spectrometer: Perkin-Elmer 521, range: ItOOO - 650 cm \ Spectra driven in Nujol (peaks caused by Nujol are not included); S = strong, sh = shoulder, m = moderate and v = weak, Characteristic peaks are marked with an asterisk (*).

Form I

65623 3520 v * 1590 s 1262 m 100¾ m 3460 m 1556 m 121 * 8 m 978 m 3370 m 153¾ s 1170 m 918 v 3265 s 1480 m 1152 m 882 s 1670 sh 11 + 10 s 113I + w 878 sh 1752 s 1UOO s 1112 m 838 w 1706 s 1338 s 1076 m 81¾ w 1660 s 1328 s 1054 sh 790 v 1620 s 1284 m * 1042 s 778 v 754 m

Form II

3526 m 1544 s 1152 m 920 v 3492 v 1478 m 1142 m 882 m * 3364 m 1412 s 1112 m 878 w 3250 m 1398 s 1080 m 840 v 1758 s 1332 s 1058 m 818 v 1695 s 1284 m * 1045 m 792 v 1665 s 1268 m 1005 m 752 m 1642 sh 1240 m 980 m 1624 s 1172 m 954 v

Form III

3465 m 1532 s 1155 m 820 v 3415 m 1482 m * 1124 s 800 v 3345 m 1412 s 1078 m 790 v 3275 m 1395 s * 1058 s 768 m 3200 m 1326 s 1048 s 748 m 1780 s 1285 m 1014 m 1702 s 1260 m 985 m 1660 s 1230 m 938 v 1632 m 1225 m 888 ra 1618 s 1196 w 878 s 1552 m 1180 m 836 v

Form IV

10 65623 3585 w 159¼ s 126U m 1008 m 788 w 3520 v 1555 m 12k0 m 980 m 752 m 3370 m 15 ^ 0 m 1172 v 956 w * 3260 s 11 * 78 m 1152 m 920 v 1758 s 11 * 10 s 1111 * w 882 m 1712 s 11 * 00 s 1078 m 878 v 1661+ s 1330 s 1058 m 838 w 1620 s 1285 m 10l * 2 m 818 v

If insoluble salts of a compound of formula I are desired for a particular purpose, e.g. in sustained release preparations, such salts may be prepared in a conventional manner, e.g. with suitable organic amines.

The compound of formula I is prepared analogously to methods known per se such as. As mentioned above, hydrogen or a carboxyl-protecting group, i.e. e.g. a residue of an ester-forming aliphatic or araliphatic alcohol or an ester-forming phenol, silanol or stannanol, or a symmetrical or mixed anhydride group derived from a suitable acid, and R is a hydrogen-protecting N-protecting group, i.e. e.g. an acyl group, in particular a lower alkanoyl group, e.g. acetyl, a halogen-substituted lower alkanoyl group, e.g. a mono-, di- or trichloroacetyl or chlorosulphonyl group. The acid addition salt may be formed, for example, with a mineral acid such as hydrochloric, hydrobromic, sulfuric, nitric or phosphoric acid, or with an organic acid such as methanesulfonic or toluene-p-sulfonic acid. Base salts can be prepared by reacting cephalosporinic acid with sodium or potassium 2-ethylhexanoate.

The compound of formula I can most conveniently be prepared by condensing a compound of formula II with an acylating agent, such as an acid halide of an acid of formula III, especially an acid chloride or bromide. This acylation can be carried out at a temperature of -50 ° to + 50 ° C, preferably -20 ° to + 30 ° C. The acylation can be performed in an aqueous or non-aqueous medium.

The acylation can be carried out in the presence of an acid scavenger, e.g. a tertiary amine such as triethylamine or dimethylaniline, an inorganic base e.g.

The free acid form of the compound of formula III can be used as such as an acylating agent. Most preferably, such acylations are carried out, for example, with a carbodiimide such as N, N'-diethyl, dipropyl or diisopropylcarbodiimide, N, N'-dicyclohexylcarbodiimide, N-ethyl-N1-n-dimethylaminopropylcarbodiimide, in the presence of a carbonyl compound, e.g. carbonyldiimidazole or an isoxazolinium salt, e.g. N-ethyl-5-phenylisoxazolinium 3'-sulfonate or N-tert-butyl-5-methylisoxazolinium perchlorate. Preferably, the condensation reaction is carried out in an anhydrous reaction medium, e.g. methylene chloride, dimethylformamide or acetonitrile.

The acylation may also be carried out with a derivative of the free amide of the free acid of formula III, e.g. a symmetrical anhydride or a mixed anhydride such as pivalic acid, or a halogen formate, e.g. a lower alkyl haloformate. Mixed or symmetrical anhydrides can be formed during the reaction. Thus, for example, a mixed anhydride can be prepared using N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline. Mixed anhydrides can also be prepared with phosphoric acids, e.g. phosphoric acid or phosphoric acid, sulfuric acid or aliphatic or aromatic sulfonic acids, e.g. p-toluenesulfonic acid.

When using a starting material of formula IV, suitable carbamoylating agents are those of formula R13-NCO (V) • ·. 13 isocyanates wherein R is a leaving group; these carbamoylating agents give a protected carbamoyloxymethyl group of the formula -ch20-co-nhr13 10 in the 3-position, in which formula R has the same meaning as above, which group can be converted into the desired unsubstituted 3-carba-. . 13. ....

to a moyloxymethyl group from which the R group can be cleaved, for example,

lysing. Suitable R groups which are readily cleavable by such work-up include, for example, chlorosulfonyl and bromosulfonyl, aralkyl such as benzyl, p-methoxybenzyl and diphenylmethyl, tert-butyl, halogenated lower alkanoyl such as trichloroacetyl and dichloroacetyl, and halogenated alicarboxylated , 2,2-trichloroethoxycarbonyl.

Such R groups (with the exception of aralkyl groups, e.g. diphenylmethyl) can generally be cleaved by acid or base catalyzed hydrolysis, for example by base catalyzed hydrolysis using sodium bicarbonate. Halogenated groups, e.g. chlorosulfonyl, trichloroacetyl and 2,2,2-trichloroethoxycarbonyl, can also be cleaved by reduction, and for example chloroacetyl can be cleaved by treatment with thioamide, for example thiourea. Aralkyl groups, for example diphenylmethyl, can be easily cleaved by treatment with an acid such as a strong organic acid, for example trifluoroacetic acid.

Preferably an excess of the carbamoylating agent of the formula V is used, e.g. at least 1.1 mol / l of the compound of the formula IV. Carbamoylation can be performed in the presence of a base, for example a tertiary organic base such as tri (lower alkyl) amine, for example triethylamine, or using an alkali metal salt of an acid of formula IV, for example the sodium salt, although such an excipient is not required for more active isocyanates, for example. . . . . 13.

for compounds of formula V wherein R is a highly electron repellent group, e.g. chlorosulphonyl or trichloroacetyl. Thus, carbamoylation, in which an excess of free acid (IV) and isocyanate (V) is reacted, 13 .... .....

wherein R is, for example, a chlorosulfonyl or triclonacetyl group, is particularly preferred in practice under simple reaction conditions because the carboxy group in the Π-position of the cephalosporin does not need to be temporarily protected and then deprotected, and because the N-protected 3-carbamoyloxy formed. .... .13. .

the electron-repellent R group of the methyl cephalosporin product can be easily removed, e.g., by hydrolysis with aqueous sodium bicarbonate solution.

It should be noted that it may be convenient to retain or even add N-substituents. ...

introducing an R J group during changes to the 3-carbamoyloxymethyl intermediates to reduce undesired side reactions associated with the carbamoyloxymethyl group.

Another useful carbamoylating agent is cyanic acid, which is most conveniently prepared in the reaction from e.g. an alkali metal cyanate such as sodium cyanate. The reaction is carried out in the presence of an acid, for example a strong organic acid such as trifluoroacetic acid. Cyanic acid effectively corresponds to a compound of formula V wherein R is hydrogen and thus directly converts compounds of formula IV to 3-carbamoyloxymethyl derivatives.

The 3-hydroxymethyl starting material used in the process of the invention can be prepared, for example, by the methods described in GB Patent No. 1,121,308 and BE Patent No. 783 * + * + 9.

The substituent blocking group for the U-carboxy group of the compounds of formula II or IV is preferably one which is easily cleaved at a later stage of the reaction steps and preferably contains 1 to 20 carbon atoms. Suitable carboxyl blocking groups are e.g. a list of groups well known and representative in the art is contained in the aforementioned BE Patent No. 78 * + UU9. Preferred carboxylic blocking groups are aryl-lower alkoxycarbonyl groups, e.g. p-methoxy-13,65623 benzyloxycarbonyl, p-nitrobenzyloxycarbonyl and diphenylmethoxycarbonyl, lower alkoxycarbonyl groups, e.g.

The carboxyl blocking group can be subsequently removed by suitable methods mentioned in the literature. Thus, in many cases, acid- or base-catalyzed hydrolysis as well as enzyme-catalyzed hydrolysis can be used. Results of Therapeutic Experiments The antibacterial activity (lowest effective concentration) of the compound (syn-isomer) prepared according to the invention was determined against both normal microorganisms and resistant strains. 3- (2-Chloroethylcarbamoyloxymethyl) -7β- (2-methoxyiminophenylacetamido) -cef-3-em-1-carboxylic acid (Example 1363/72, Example 53) was used as a reference · The results are shown in the following table .

Microorganism Lowest effective concentration (pg / ml) (6R, 7R) -3-carbamoyl-3- (2-chloroethyl-7- / 2- (fur-2-yl) -2-carbamoyloxymethyl) methoxyiminoacetamido7- 7β- (2-Methoxyimino-cef-3-em-U-carboxyl-phenylacetamido) -cefic acid (syn-isomer) 3-em-U-carboxylic acid

Staph, aureus 6θ * + Ε 0.5 1.25

Staph, aureus 853E 0.5 1

Staph, aureus 1 ^ + 1 i + E 0.5 1

Staph, aureus 663E 0.2 1.25 E. coli (TEM +) 1193E b 8 E. coli 573E b 8 S. typhimurium 8ol + E 1+ 16

PR. mirabilis 1 + 31E 1 8

PR. morganii 235 0.25 1+ H. influenzae 1151 + E <0.5 16

The table shows that the new antibiotic is more active than the compound known from Finnish application 1363/71 +. It is more active, especially against gram-negative microorganisms.

The compound according to the invention can be used as suspensions, solutions or emulsions in an oily or aqueous medium and can be mixed with suspending, stabilizing and / or dispersing agents. Alternatively, the active ingredient may be in powder form for constitution with a suitable vehicle, e.g. sterile, pyrogen-free water.

lb 65623

For veterinary use, the compositions may take the form of, for example, long-acting or rapid-release preparations.

In general, the compositions will contain from 0.1 to 99% »preferably from 10 to 60% of active ingredient, depending on the route of administration. When the compositions are in unit dosage form, each unit will preferably contain from 50 to 1500 mg of active ingredient. The treatment dose for an adult preferably contains 500-1 * 000 mg per day, depending on the route and frequency of administration.

The compounds of the invention may be administered with other compatible drugs such as penicillins, other cephalosporins or tetracyclines.

The following examples illustrate the invention. All temperatures are in degrees Celsius. Melting points were determined on a Kofler instrument.

A. Preparation of starting material

Preparation 1

Diphenylmethyl (6R, 7R) -7-amino-3-karbamoyylioksiraetyyli-ceph-3-em-L-karboksy1aattitolueeni-p-sulfonic acid salt.

A stirred suspension of 156 g (0.75 moles) of phosphorus pentachloride in 1.5 liters of dry dichloromethane was cooled in an ice bath and treated with 60.5 ml (0.75 moles) of pyridine at a rate to maintain the temperature of the mixture at about 20 ° -25 The mixture was stirred and cooled to 8 ° C and 35.5 g (0.5 moles) of diphenylmethyl- (6R, 7R) -7- (thien-2-yl) -acetamido was added portionwise over 10 minutes. -3-Trichloroacetylcarbamoyloxymethyl-cef-3-emA-carboxylate. The mixture was stirred for 13 hours at about 8 ° C, then a mixture of 225 ml (2.5 mol) of butane-1,3-diol and 500 ml of dichloromethane, pre-cooled to 20 ° C and stirred, was added over 10 minutes. , keeping the temperature of the mixture in the range of -15 ° to -20 ° C. The cooling bath was removed and the mixture was stirred for 20 minutes at about -10 ° C. One liter of water was added and the 65,623 biphasic mixture was stirred for 30 minutes. The aqueous phase was extracted with 2 x 500 ml of dichloromethane, the organic phases were washed successively with 1 liter of 2N hydrochloric acid, combined and evaporated to a brown gum. The gum was dissolved in 3.6 liters of methanol and this solution was stirred for 1 1/2 hours at about 20 ° C and a small amount of brown solid was removed by filtration. The yellow filtrate was concentrated in vacuo (bath temperature not exceeding U0 ° C) to 1.5 liters and 1.5 liters of water were added. The resulting suspension was cooled for one hour, the yellow solid was isolated by filtration, washed well with water, sucked as dry as possible and dried overnight in vacuo at 1 + 0 ° C. The fatty solid thus obtained and 81 g (0.25 mol) of toluene p-sulfonic acid monohydrate were added to two liters of stirred chloroform. After several minutes, the toluene-p-sulfonic acid salt began to crystallize. Stirring was continued for 30 minutes after which the water was removed azeotropically in vacuo by continuous replacement of chloroform so that the volume remained at 2 liters, the suspension was cooled overnight and the product was filtered off, washed with 2 x 250 ml chloroform, filtered again, washed with 250 ml chloroform and dried in vacuo at 1 ° C to give 237.8 g (7.1%) of the title compound as an off-white crystalline solid, = 262 nm (ε = 7250), NMR analysis of ZQcmCS f (dimethyl sulfoxide) showed that 0.25 moles of chloroform was present.

Preparation 2 (6R, 7R) -7-Amino-3-carbamoyloxymethyl-ceph-3-em-U-carboxylic acid 300.0 g (0.1 U moles) of (6R, 7R) -7-7-mole were added portionwise over 30 minutes. amino-3-carbamoyloxymethyl-ceph-3-em-1 + -carboxylate toluene-p-sulfonic acid salt solvated with about 0.6 moles of chloroform in a stirred mixture of 300 ml of trifluoroacetic acid and 300 ml of anisole in a water bath at 20 ° C: in. During the first 20 minutes, the temperature rose from 23 ° C to 28 ° C, but decreased to 26 ° C at the end of the addition. The golden solution was stirred for one hour as the temperature dropped to 21 ° C and then added to a stirred mixture of 1.5 L of ethyl acetate and 1.5 L of water immersed in an ice bath. The pH of the stirred mixture was adjusted to 3.8 over 10 minutes with ammonia solution (density 0.880) as the temperature rose to 38 ° C. The suspension was stirred and cooled to 10 ° C for 1 1 / U and filtered. The cream solid was washed with 750 ml of water and hx with 200 ml of ethyl acetate and dried in vacuo to give 115.6 g (96.2%) of the title compound, (pH = 6-phosphate buffer) = 265 nm (λ = 7750), purity by high pressure liquid chromatography 99.7% · Microanalytical values confirmed the structure of the title compound.

16 65623

Preparation 3 a) (6R, 7R) -7- (R-5-Benzoylamino-5-carboxypentanamido) -3-hydroxymethyl-cef-3-em-4-carboxylic acid A stirred solution was cooled to + 5 ° C where was 62.00 g (ca. 100 mmol) of the monopotassium salt of (6R, 7R) - * 7- (R-5-amino-5-carboxypentanamido-3-hydroxymethyl-cef-3-em-k-carboxylic acid (purity n, 67 %) In 300 ml of water and treated for 25 minutes with a solution of 17s1 * ml (150 mmol) of benzoyl chloride in 200 ml of acetone. The pH of the reaction mixture was adjusted to 8.2-8.5 by the controlled addition of 30 w / v. The mixture was stirred for a further 10 minutes, 10 ml of ethyl acetate was poured on and the pH was lowered to 5 * 6 with orthophosphoric acid, the layers were separated and the aqueous layer was washed with 2 x 1 + 00 ml of ethyl acetate, the aqueous layer was diluted with 2 liters of water and The pH of the mixture was adjusted to 2.0 with orthophosphoric acid, the layers were separated and the aqueous layer was extracted with 2 x 1500 mL of ethyl acetate. etaattia. The combined extracts were washed with 800 mL of saturated brine, dried and concentrated in vacuo to a volume of 300-00 mL. The resulting suspension was stirred for 20 minutes in two liters of ether and filtered. The isolated solid was washed with 2 x 250 mL of ether and dried in vacuo (Qtorr) to give 5.95 g (88.8 ρ / ρ- / ί), ΟΛ _ = (C = 1.00 in dioxane), y \ mgQts (pH = 6-buffer) = 231 nm (E1cm = 2 ^ 5) 2 && mi (inflation, = 1 i + 5), NMR analysis (dimethyl sulfoxide-dg) showed the presence of about 20% lactone and n .0 U moles of ethyl acetate.

b) (6R, 7R) -7- (R-5-Benzoylamino-5-carboxypentanamido) -3-chloro-acetylcarbamoyloxymethyl-ceph-3-em-1 + -carboxylic acid monosodium salt 25.6 g of the above a) were treated. product with a solution of 9.00 g (75 mmol) of chloroacetyl isocyanate in 92 ml of dry acetone. The resulting solution was stirred for 25 minutes at about 20 ° C, cooled to about 5 ° C over 5 minutes and treated with a solution of 8.1 + 7 g (51 mmol) of sodium 2-ethylhexanoate in 51 ml of acetone. . The crystalline suspension was stirred for 5 minutes at about 5 ° C, the solid was filtered off, washed with 80 ml of acetone and 250 ml of ether and dried in vacuo (itorr) to give 27.23 g (107.0 rpm). —JS) of the title compound, λ <= + 72.0 ° (c = 1.00 in 3 # aqueous NaHCO 3), J (pH = 6 buffer) = 227 mm (E. = 2k9), 261 nm (inflation, R- | cm - 105). NMR analysis (dimethyl sulfoxide-d 6) showed the presence of n, 35% lactone and about 1.0 mole of chloroacetamide as an impurity.

c) (6R, 7R) -7-Amino-3 "chloroacetylcarbamoyloxymethyl-cef-3-em-k-carboxylic acid π 65623 Cooled with stirring to about 10% of a suspension of 21 *, 77 g of the product from h) above. In 320 ml of dry methylene chloride under nitrogen, 17.60 ml (218.0 mmol) and then 16.80 ml (139.2 mmol) of dichlorodimethylsilane and a light brown suspension were added and the mixture was stirred for 20 minutes.

At 20 ° C and then cooled to -17 ° C. 10.81+ g (52.0 mmol) of phosphorus pentachloride were added and the mixture was stirred for two hours at -17 ° C to 23 ° C.

6.1 + 8 ml (80.1+ mmol) of pyridine were added and the mixture was added to 10 ml + methanol and 20 ml of washing liquid pre-cooled to -35 ° C at such a rate that the temperature of the stirred mixture did not exceed -10 ° C. The stirred solution was allowed to reach + 2 ° C over 25 minutes, then 88 ml of water was added and the pH of the mixture was adjusted from 0.6 to 3.8 with aqueous ammonia solution (density 0.880). The biphasic mixture containing the precipitate formed was cooled for one hour and then filtered. The solid was washed successively with 100 mL of 50 v / v aqueous methanol, 80 mL of methanol and 1 + 0 mL of methylene chloride and dried in vacuo (1 torr) to give 6.86 g (27.7 g). p / p- #) title compound as a cream powder, Γ ° * Ό ^ = + * + 8 ° (c - 1.01 * in dimethyl sulfoxide), (pH * 6 buffer) = 237.5 nm (Emax = Λ max 9), 261.5 nm (λ max = λ max).

B. Examples

Example 1 a) Diphenylmethyl (6R, 7R) -3-carboamoyloxymethyl 7- [2- (fur-2-yl) -2-methoxyiminoacetamide] cef-3-em-1-carboxylate (syn-isomer)

Method I

25.0 g (0.33 mol) of crude diphenylmethyl- (6R, 7R) -7-amino-3'-carbamoyloxymethylcef-3-em-1 + carboxylate toluene-p-sulfonic acid salt obtained from the corresponding 3-trichloroacetylcarbamoyloxyacetyl compound were dissolved. to a mixture of ethyl acetate and aqueous sodium bicarbonate. The organic layer was separated, washed with water, dried over magnesium sulfate and evaporated on a rotary evaporator to give 11.5 g (0.262 mol, 77%) of diphenylmethyl (6r, 7R) -7-amino-3-carbamoyloxymethylcef-3-em-1 + carboxylate. .

5.32 g (0.312 moles) of 2- (fur-2-yl) -2-methoxyiminoacetic acid (syn-isomer) in 100 ml of dry dichloromethane were added to 50 ml of this amine cooled to 3 ° C and 10 ml of dichloromethane. minutes later 6.5 g (0.312 mol) of DL-dicyclohexylcarbodiimide in 30 ml of dichloromethane. The reaction mixture was stirred in an ice bath for 1 + 5 minutes during which time a solid (probably N, N'-dicyclohexylurea) crystallized. The precipitate was filtered off and discarded, the filtrate was washed with aqueous sodium bicarbonate solution and water, dried over magnesium sulphate and evaporated to dryness. The residue was triturated in ethanol to give 10.6 g of crude product, which was purified by chromatography on one kilogram of silica gel. Elution with 10- $ acetone in dichloromethane removed the non-polar impurities and fractions eluted with 20- $ acetol in dichloromethane gave h-, 8 g ($ 31) of the title compound, m.p. 199-202 ° C, Z ~ o7p1 ~ + 11 + 0 (c = 1.0 dimethyl sulfoxide),.

IR, NMR and microanalytical values confirmed the structure of the title compound.

Method H

1.86 g (18.4 mmol) of triethylamine were added to a solution of 3.1 g (18.1+ mmol) of 2- (fur-2-yl) -2-methoxyiminoacetic acid (syn-isomer) in 35 ml: in dichloromethane. After cooling this solution in an ice bath for 5 minutes, 1.57 ml (18 U mmol) of oxalyl chloride and a drop of N, N-dimethylformoform were added. After half an hour the solvent was removed in vacuo and the solid residue was dried in vacuo one hour. 150 ml of anhydrous ether was added to dissolve the acid chloride formed, and 2.5 g of insoluble triethylamine hydrochloride was removed by filtration. The ether was evaporated on a rotary evaporator and the oily residue was redissolved in dichloromethane.

8.9 g of (1-7 moles) of diphenylacetyl- (6R, 7R) -7-amino-3-carbamoyloxymethyl-cef-3-em-carboxylate toluene-p-sulfonic acid salt were dissolved in anhydrous dichloromethane. This solution and aqueous sodium bicarbonate were shaken, washed with water and dried over magnesium sulfate. To this solution of the free amine was added a solution of 2- (fur-2-yl) -2-methoxyiminoacetyl chloride (syn-isomer) in dichloromethane and 5 ml of propylene oxide. After 10 minutes, 1.1 g of a crystalline solid was isolated by filtration, which was later identified as diphenylmethyl (6R, 7R) -7-amino-3-carbamoyloxymethyl-cef-3-em-U-carboxylate hydrochloride. The filtrate was washed with 2N sulfuric acid, water, aqueous sodium bicarbonate and water, dried over magnesium sulfate and evaporated to dryness to give 2.5 g (30.5%) of the title compound having the same physical properties as the product of method (I) above.

b) Sodium (6R, 7R) -3-carbamoyloxymethyl 7- [2- (fur-2-yl) -2-methoxyiminoacetamide] cef-3-em-1 + carboxylate (syn-isomer)

20 ml of trifluoroacetic acid were slowly added to a mixture of 5 ml of anisole and 0.7 g (8 mmol) of diphenylmethyl- (6R, 7R) -3-carbamoyloxymethyl-7- [2- (fur-2-yl) -2-methoxy- iminoacetamido-7-ceph-3em-4-carboxylate (syn-isomer) cooled in an ice bath. The flask was shaken occasionally over the next 10 minutes to completely dissolve the solid. The flask was removed from the ice bath and the excess trifluoroacetic acid was removed on a rotary evaporator. Trituration of the residue in 5 ml of ethyl acetate gave 3.3 g (9%) of (6R, 7R) -1965623 3-carbamoyloxymethyl-7- [2- (fur-2-yl) -2-methoxyiminoacetamide] -cef -3-em-1 + -carboxylic acid (syn-isomer) as a solid which was isolated by filtration and washed with diethyl ether.

The free acid was dissolved in acetone and 8.0 ml of sodium 2-ethyl was added. . . . . . o 1-m acetone solution of hexanoate. After the reaction mixture was stirred at 0 ° C for two hours, 2.3 g (73%) of the title salt were filtered off. This and 0.8 g of the second batch of the title salt were combined and dissolved in 250 ml of water and extracted twice with 100 ml and once with 50 ml of ether. The aqueous solution was lyophilized to give 2.66 g of sodium (6R, 7R) -3-carbamoyloxymethyl-7- [2- (fur-2-yl) -2-methoxyiminoacetamido] cef-3-em-1 + - carboxylate (syn isomer), λ <Xj '= 73.5 (c = 1.06 in dimethyl sulfoxide), λmax = 27 * + nra = 500), γ (Nujol) 31 + 50, 3330, 3250 (NH, NHg and H 2 O), 1752 (azedin-2-one), 1652 and 1600 cm -1 (carboxylate), 1 C (dimethyl sulfoxide-dg) 0.2l + (d, J = 8 Hz, CONH), 2.12 (d, J = 2 Hz, furyl C 1 H -H), 3.25 and 3.30 (m, furyl C ), 1 +, 32 (dd, J = 5 and 8 Hz, C Ha, C3-CH2), 6.07 (s, NOCH2 and 6.58 (q, J = 18 Hz, <^ 0.

Analysis for CH 2 Cl 2 / Na 2 O 2 SO 4. For 5H 2 O (mp 1 + 55.37)

Calculated: C 1 + 2.2 H 3.5 N 12.3 S 7%

Found: C 1 + 2.0 H 3.8 N 12.1 S 7.2%

Example 2 (6R, 7R) -3-Carbamoyloxymethyl-7- [2- (fur-2-yl) -2-methoxyiminoacetamide] cef-3-era-1-carboxylic acid (syn-isomer)

A stirred mixture of 75 ml of N, N-dimethylacetamide, 75 ml of acetonitrile, 1 + 2 ml (0.3 mol) of triethylamine a and 16.1 + 0 g (0.06 mol) of (6R, 7R) 7-Amino-3-carbamoyloxymethylcef-3-em-1-carboxylic acid was immersed in an ice bath and 10 ml of water were added. The mixture was stirred at 0 -2 ° C for 1 + 5 minutes and the solid slowly dissolved to form a yellow solution,

At the same time, a stirred suspension of 11 +, 99 g (0.072 mol) of phosphorus pentachloride in 150 ml of dry dichloromethane was cooled to 0 ° C and 27.5 ml of Ν, Ν-dimethylacetamide were added. The resulting solution was recooled to -10 ° C and 12.17 g (0.072 mol) of 2- (fur-2-yl) -2-methoxyiminoacetic acid (syn isomer) were added. The mixture was stirred for 15 minutes at -10 ° C and 35 g of crushed ice was added. The mixture was stirred for 10 minutes at 0 ° C and the lower dichloromethane phase was added over 10 minutes to the cephalosporin solution prepared above and cooled to -10 ° C after which the reaction temperature rose steadily to 0 ° C. The mixture was stirred for 1 hour at 0 ° -2 ° C, the cooling bath was removed and the reaction temperature was allowed to rise to 20 ° C + per hour, then the reaction mixture was slowly added at 5 ° C to 100 ml of 2N hydrochloric acid diluted with 1.15 liters. cold water. The pH of the biphasic mixture was adjusted to less than 2 with 10 mL of 2N hydrochloric acid and the mixture was stirred and cooled to 5 ° C. The precipitate was filtered, washed with 100 ml of dichloromethane and 250 ml of water and dried under vacuum at + 0 ° C overnight to give 22.0 g (86.6%) of the title compound, + 58 ° ( c = 1.08 in dimethyl sulfoxide), XmaJcs = (pH = 6-phosphate buffer) 2lh nm (& = 17500), = (Nujol) 3I + 80, 31 + 1 + 0, 336 ?, 3255 and 3133 (bound NH and NH 4), 2725 and 2590 (CO 2 H) 1760 (azetidin-2-one), 1728, 1712 and 1698 (OCONH 2 and CO 2 H), 1655 and 1530 cm -1 (CONH), T (dimethyl sulfoxide-d 6), 0.25 (d, J = 8 Hz, CONH), 2.18 (s, furyl C 1 H 3, 3.28 and 3.1+ (m, furyl C 1 H and C 3 H) 1 + 2 (s, CONH2), 1 +, 19 (dd, J = 8 and 5 Hz, C2 H), 1 +, 80 (d, J * 5 Hz, C8-H) 5.06 and 5, 39 (q, J = 13 Hz, C 3 -CH 2), 6.09 (s, NOCH 2), 6.1 + 1 + (collapsed q, ^ 2 ~ Η 2) and 7.99 (0.03 moles of CH 3 CO 2 ( CH 3) 2).

Example 3 a) (6R, 7R) -7- (R-5-Benzoylamino-5-carboxypentanamido) -3-hydroxymethyl-cef-3-em-1-carboxylic acid monoquinolinium acid monohydrate A stirred solution was cooled to 0-5 ° C. containing 18.1 + 5 g (30 mmol) of (6R, 7R) -7- (R-5-amino-5-carboxypentanamido) -3-hydroxymethylcef-3-em-1-carboxylic acid monopotassium salt in 93 ml of water ( ice-water bath) and treated for 25 minutes with a solution of 5.19 ml (1 + 5 mmol) of benzoyl chloride in 63 ml of acetone. The pH of the reaction mixture was adjusted to 8.5 (-1.1) by the controlled addition of about 100 ml of a 30 w / v aqueous solution of tricalcium orthophosphate. The mixture was stirred for a further five minutes, poured onto 150 ml of ethyl acetate and the pH was lowered to 5.6 with orthophosphoric acid. The layers were separated and the aqueous layer was washed with 2 x 300 mL of ethyl acetate. The combined washings were extracted with 200 ml of water. The combined aqueous layer and washings were diluted with 600 mL of water, 600 mL of ethyl acetate was added, and the pH of the stirred mixture was adjusted to 2.0 with orthophosphoric acid. The organic layer was separated and 10.61+ ml (1 + 5 mmol) of quinoline in 25 ml of ethyl acetate was added with stirring to give a white precipitate. The aqueous layer was extracted with 3 x 300 mL of ethyl acetate and the washings were added to the quinoline-containing suspension. The mixture was stirred for one hour at about 18 ° C and concentrated in vacuo to about 500 mL. 900 ml of ether were added with stirring and after 30 minutes the solid was isolated by filtration, washed with 5 x 200 ml of ether and dried in vacuo (1 torr) to give 18 ° 19.20 g (101 +, 1 w / w) of the title compound. as a white powder, / <* 7 = +78 1¾, (pH = 6 buffer) = 258 nm (inflation, E. = 185) - D16LKS · 1 cm IR and NMR analysis showed that the title compound contained about 15% as an impurity lactone and traces of ether and ethyl acetate.

b) (6R, 7R) -7- (R-5-Benzoylamino-5-carboxypentanamido) -3-trichloroacetylcarbamoyloxymethyl-ceph-3-em-1 + -carboxylic acid monoquinolinium salt Treated 1 +, 21 + g (7 mmol) above described product a) 100 ml of dry dioxane in which it was partially dissolved. To the stirred mixture was added 2.90 mL of 21,656,23 (2 + +, 5 mmol) trichloroacetyl isocyanate. The resulting solution was stirred for 30 minutes, clarified by filtration and evaporated in vacuo to give a yellow foam. This was dissolved in about 10 ml of acetone and poured into about 100 ml of stirred isopropyl ether. The white precipitate formed was filtered off and dried in vacuo (1 torr) to give 6.26 g (1 * + 7.8 w / w) of the title compound as a white powder. NMR analysis showed the presence of about 22% lactone, 0.75 moles of isopropyl ether, 0.2 moles of dioxane and a small amount of acetone.

c) (6R, 7R) -7-Amino-3-trichloroacetylcarbamoyloxymethyl-cef-3-em-1-carboxylic acid Cooled under nitrogen to about 10 ° C with stirring 1 +, 77 g (6 mmol) of the above product b) 1 + 0 in ml of dry methylene chloride. 2.20 ml (27.3 mmol) of pyridine and then 2.10 ml (17.1+ mmol) of dichlorodimethylsilane were added, the brown solution was stirred at about 17 ° C for 20 minutes and then cooled to -17 ° C. . 1.355 g (6.5 mmol) of phosphorus pentachloride were added and the mixture was stirred for two hours at about -16 ° C. 0.81 ml (10 mmol) of pyridine was added and the mixture was added to 13 ml cooled to -35 ° C. methanol and 2.5 ml of washing liquids at such a rate that the temperature of the stirred mixture did not exceed -10 ° C. The stirred solution was allowed to reach + 9 ° C over 25 minutes, 11 ml of water was added and the pH of the mixture was adjusted to 0.3-3.8 with ammonia solution (density 0.880). The biphasic mixture containing the precipitate formed was cooled for 1 hour and then filtered. The solid was washed successively with 12 mL of 50% w / w aqueous methanol, 10 mL of methanol and 5 mL of methylene chloride and dried in vacuo (1 torr) to give 1.22 g (25.6 w / w). ) the title compound as a cream powder = + J + 1 + 0 (c = 1.02 in dimethyl sulfoxide), "1, λ of ^ λ of Λ niQiCS.

(pH = 6-buffer) = 2 ^ + 0 nm (E. = 133), 263 nm (E, = 1 i + 0). NMR values confirmed the structure of the title compound i cm j cm.

d) (6R, 7R) -3-Carbamoyloxymethyl-7- [5- (fur-2-yl) -2-methoxyiminoacetamide] cef-3-em-1-carboxylic acid (syn isomer)

1.5 g (21.5 mmol) of phosphorus pentachloride were dissolved in 90 ml of dry methylene chloride and cooled to -15 ° C with stirring. 9 ml of Ν, Ν-dimethylacetamide were slowly added keeping the temperature below -10 ° C and the mixture was stirred for 10 minutes. 3.66 g (21.5 mmol) of 2- (fur-2-yl) -2-methoxyiminoacetic acid (syn isomer) were added and the mixture was stirred for 15 minutes at -15 ° C. 18 g of crushed ice was carefully added so that the temperature of the mixture did not exceed -7 ° C. The mixture was stirred for 10 minutes, the organic layer was isolated, added dropwise to a solution of 7.52 g (18 mmol) of (6R, 7R) -7-amino-3-trichloroacetyl-carbamoyloxymethyl-ceph-3-em-1 + -carboxylic acid -10 22 ° C in 22 ml of dry methylene chloride and 5.5 ml (10 mmol) of triethylamine. The acid chloride solution was added over 20 minutes and the temperature of the reaction mixture was maintained at -10 to -8 ° C. The mixture was then stirred for 80 minutes, allowing the temperature to rise to + 3 ° C, and 6 ml of methanol were added. After stirring for an additional 5 minutes, the solution was extracted with 2 x 120 mL of 3% w / w sodium bicarbonate and 150 mL of water. The combined extracts were allowed to stand for 3 1/2 hours at about 20 ° C, washed with 100 ml of ethyl acetate and acidified to pH 1.5 with concentrated hydrochloric acid. The resulting precipitated oil was extracted with 2 x 300 mL of ethyl acetate. The combined organic extracts were washed with 2 x 100 ml of water, dried over magnesium sulphate and evaporated in vacuo to give 7 * 1 g of a yellow solid which was stirred in 150 ml of ether, filtered and dried in vacuo (1 torr) (to give 5-20 g). (68.2% of theory) of the title compound as a yellow solid, λ = (pH = 6 buffer) 275 nm - df melts · (E * = 385).

1 cm

Example U

(6R, 7R) -3-Carbamoyloxymethyl-7 '[2- (fur-2-yl) -2-methoxyiminoacetamido] -ceph-3-em-1-carboxylic acid (syn isomer)

1.5 ml of a solution of Ν, Ν-dimethylacetamide cooled to -10 ° C with 750 mg (3.6 mmol) of phosphorus pentachloride in 15 ml of anhydrous dichloromethane were added. After 10 minutes, 612 mg (3, 6 mmol) 2- (fur-2-yl) -2-methoxyiminoacetic acid (syn isomer). The suspension, which soon became a clear solution, was stirred for 15 minutes at -10 ° C. 3 g of ice were added and after 10 minutes the layers were allowed to separate in a dropping funnel. The organic phase was slowly run over 5 minutes into a solution of 1.05 g (3 mmol) of (6R, 7R) -7-amino cooled to -10 ° C. - 3-chloroacetylcarbamoyloxymethylcef-3-em-1 * -carboxylic acid in 15 ml of dichloromethane with 0.9 ml (6.5 mmol) of triethylamine. After forty minutes, 1 ml of methanol was added, and five minutes later, the reaction mixture was extracted twice with 150 ml of 3% aqueous sodium bicarbonate solution. The aqueous extract was washed with 25 ml of ethyl acetate and allowed to stand for four hours at 20 ° C. The solution was washed twice with ethyl acetate, acidified with 2N hydrochloric acid and extracted three times with ethyl acetate. The combined organic layers were dried over magnesium sulfate, decolorized with activated charcoal and evaporated in vacuo to give 1.15 g (87%) of a pale yellow solid. It was washed with diethyl ether and isolated by filtration to give 0.91 g (71%) of the title compound, λ max = 27 nm (f. = 17300).

23 65623

Example 5 (6R, TR) -3-Carbamoyloxymethyl-methyl-2-yl) -2-methoxyimino-acetamide-7-ceph-3-em-U-carboxylic acid (syn-isomer) 750 ml of acetone were cooled to 0 ° C: treated with 28.8 ml (2.0 mmol) of trichloroacetyl isocyanate and the solution was recooled to 0 ° C. To the stirred isocyanate solution was added 5 5.6 g (120 mmol) of (6R, 7R) -7- [5- (fur-2-yl) -2-methoxyiminoacetamido] -3-hydroxymethyl-ceph-3-em-U-carboxylic acid. (syn isomer) in portions over five minutes so that the reaction temperature does not exceed 6 ° C. The yellow solution was stirred for 15 minutes and 1.5 mL of methanol was added. The solution was concentrated to 60 mL and the concentrate was dissolved in 750 any methanol. U5.3 g (5 + + mmol) of sodium bicarbonate in 600 ml of water and 5 g of activated carbon were added and the resulting suspension was stirred for two hours at room temperature. The charcoal was removed by filtration through diatomaceous earth and the pH of the pale yellow filtrate was adjusted to 4.5 by the addition of dilute hydrochloric acid. The solution was concentrated in vacuo to half volume and an equivalent volume of water was added. The pH was adjusted to 2.0 with dilute hydrochloric acid and the product was filtered off, washed with 3 x 150 ml of water and dried for 16 hours under vacuum and at 0 ° C to give 37 * ^ 6 g (73.5%) of the title product, ~ + 63.7 ° (c = 1.0 in 0.2-m pH = 7-phosphate buffer), (pH = 6-phosphate buffer) = 27 nm (6 = 17600).

Method (II)

A suspension of 3.81 g (9 * 55 mmol) of (6R, 7R) -7 - [- (fur-2-yl) -2-methoxyiminoacetamide] -3'-hydroxymethylcef-3-em-1 + carboxylic acid (syn isomer) in 70 ml of dichloromethane and 25 ml of tetrahydrofuran and treated at 5 ° C with 2.6 ml (25 mmol) of dichloroacetyl isocyanate. The reaction mixture was treated as in Method (i) to give 3.36 g (83.0%) of the title compound, + 63 °, λmax β 273.5 nm (t-17800).

Method (lii)

A suspension of 19.05 g (50 mmol) of (6R, 7R) -7- [2-fur-2-yl) -2-methoxyiminoacetamido] -3-hydroxymethylcef-3-em-h-carboxylic acid (syn isomer) in 250 mL of dry acetonitrile was treated at 5-10 ° C with 6.33 mL (75 mmol) of chlorosulfonyl isocyanate in 80 mL of acetonitrile.

The reaction mixture was stirred for 10 minutes at 0 -5 ° C and 50 ml of water were added. The mixture was stirred at about 20 ° C and after 20 minutes white crystals precipitated. Evaporation and filtration gave 18.17 g (85.7%) of the title compound, 2k 65623 (+ 62.5 °, λ max = 273.5 nm (ε = 17820)). Evaporation of the mother liquors gave 1.88 g (8.6¾) of a second crop of product having the same values.

Example 6

Form I Sodium (6R, 7R) -3-carbamoyl-oxymethyl 7-Z2- (i-ur-2-yl) -2-methoxyiminoacetamido-7-cef-3-em-1-carboxylate (syn-isomer)

Method (I)

100 g of (6R, 7R) -3-carbamoyloxymethyl-7- [2- (fur-2-yl) -2-methoxyiminoacetamido] -ceph-3-em-4-carboxylic acid (syn-isomer) were dissolved in a mixture of 1 + 00 ml of Ν, Ν-dimethylacetamide and 1000 ml of acetone, and bo g of sodium 2-ethylhexanoate in 200 ml of acetone were added. The mixture was seeded and stirred at room temperature for 1 1/1 + hours. The product was filtered off, washed with 500 ml of acetone, extracted with 3 x 300 ml of acetone and finally with ether to give 101.1+ g (92.5%) of pure product containing 0.65 molar equivalents of water in air after equilibration. Product values j = + 61 ° (£ = 0.5 pH = 1 +, in 5-phosphate buffer) and? 273 nm and ®1cm = 1 + 12 (H 2 O).

Method (II)

Method (i) was repeated except that the cephalosporinic acid was initially dissolved in a mixture of Ν, Ν-dimethylformamide and denatured alcohol instead of N, N-dimethylacetamide acetone to give the title compound (80%) having the same properties as the product of method (I), IR spectrum showed the product to be a Form I salt.

Method (lii)

1 +, 21 + g (10 mmol) of (6R, 7R) -3-carbamoyloxymethyl-7- [2- (fur-2-yl) -2-methoxyiminoacetamido] -ceph-3-em-U-carboxylic acid ( syn isomer) to 20 ml of Ν, Ν-dimethylacetamide, dried for 2 hours on a molecular sieve (Linde 1 + A). A solution of 2.0 g (12 mmol, recrystallized from dioxane and dried over phosphorus pentoxide sodium 2-ethylhexanoate in 80 ml of ethyl acetate dried over 2b hours on a molecular sieve (linde 1 + A) was added and the solution was stirred in a sealed vessel for about 15 g. minutes until crystallization began and cooled for 1 hour at 1 ° C. The product was filtered, washed with about 100 ml of dry ethyl acetate and then transferred to an oven and dried overnight at 20 ° C and in vacuo to give 3.89 g (87 %) of the title compound.

25 65623

Example 7

Form II Sodium (6R, 7R) -3-carbamoyloxymethyl-7 ', 2- (fur-2-yl) -2-methoxyiminoacetamido] -cef-3-em-1-carboxylate (syn-isomer)

Method (I)

0.2 g of charcoal was added to a solution of 1, 00 g (9.2 g) of (6R, 7R) -3-carbamoyloxymethyl-7- [2- (fur-2-yl) -2-methoxyiminoacetamido] -cef-3-em-1-carboxylic acid (syn isomer) in 132 ml of acetone and 1.33 ml of water. The suspension was stirred for 30 minutes, filtered through a pad of diatomaceous earth and the filter pad washed with 10 ml of acetone. To the stirred filtrate was added a filtered solution of 1.66 g (10 mmol) of sodium 2-ethylhexanoate in 20 mL of acetone over 1 hour. The resulting suspension was stirred for an additional 10 minutes, the white solid was filtered off, washed with 2 x 25 mL of acetone and dried in vacuo to give 1 +, 06 g (93.0%) of the title compound, * + 60 ° (c <0 ° C). , 91 in water), λ (water) = 27 U nm (δ = 17 λ 00).

Analysis for C 0.7 HgO (mp 1 + 59.0)

Calculated: C 1 + 1.8 H 3.6 N 12.2 Na 5.0 S 7.0 HgO 2.7 ί

Found: C 1 + 1.1 H 3.55 N 12.3 Na 5.2 S 6.7 HgO 2.7.

Purity by HPLC 99.1+% ·

Method (II)

16.98 g (1 + 0 mmol) of (6R, 7R) -3-carbamoyloxymethyl-7-Z2- (fur-2-yl) -2-methoxyiminoacetamide-7-ceph-3-em-1-carboxylic acid (syn isomer) to a stirred mixture of 333 ml of acetone and 8.5 ml of water. After treatment with charcoal and filtration of the solution, 7.32 g (1 + 1 + mmol) of sodium 2-ethylhexanoate in 85 ml of acetone were slowly added over the course of one hour. The reaction mixture was stirred for 15 minutes, filtered, the product washed with 2 x 65 ml of acetone and dried overnight in vacuo at 20 ° C to give 17.95 g (98.5%) of the title compound containing 0.5 mol of water.

Example 8

Form III Sodium (6R, 7R) -3-carbamoyloxymethyl-7-Z2- (fur-2-yl) -2-methoxyiminoacetamido-7-ceph-3-em-1-carboxylate (syn-isomer)

Dissolved 1 *, 0 g of sodium (6R, 7R) -3-carbamoyloxymethyl-7 ', 2- (fur-2-yl) -2-methoxyiminoacetamido] cef-3-em-1-carboxylate (syn isomer) To 20 ml of water. 20 ml of technical grade, denatured alcohol and 160 ml of 26,656,23 dioxane were added, the solution was filtered and allowed to crystallize at + 1 ° C. The very white, needle-like crystals were filtered off, washed with 100 ml of dioxane and transferred to a oven as dioxane wet and dried overnight at 20 ° C and under vacuum to give 3.96 g (78.5%) of the title compound.

Example 9

Form IV Sodium (6R, 7R) -3-carbamoyloxymethyl-7- [2- (fur-2-yl) -2-methoxyiminoacetamide] cef-3-em-carboxylate (syn-isomer)

Sample of Form 3 and Form III sodium 3-carbamoyloxymethyl 7- [2- (fur-2-yl) -2-methoxyiminoacetate] -ceph-3-em-U-carboxylate (syn-isomer) prepared according to Example 6 according to Method III and Example 8, was placed in a humid environment ($ 75 relative humidity) for three days to give the title compound. Karl-Fischer water analysis gave U, 0j and 3.85¾, respectively (1 mole of water corresponds to 3.9Ä).

Claims (4)

A process for preparing an antibiotically active (6R, 7R) -3-carbamoyloxymethyl-7- [2- (fur-2-yl) -2-methoxyiminoacetamide] cef-3-em-1-carboxylic acid having the formula 27 62 3 HH ^ 1 -C-CO-NH - N ^ 0 </ 1-N s? -CH O-CO-NH (I) N o J och 3 COOH which compound is the syn isomer or at least 90% of the syn as an isomer-containing mixture of syn and anti-isomers, and for the preparation of its non-toxic salts, characterized in that a) condensation according to the general formula HH ΐ j / S yi — Γ Ί JN -CH2O-CO-NHR12 (II) COOR11 11 12 a compound of the formula wherein R is hydrogen or a carboxyl protecting group, and R is hydrogen or an N-protecting group, or an acid addition salt thereof, with an acylating agent corresponding to an acid of the formula? -C-C00H (III) ^ OCHg, or b) is administered 28H ~~~ \: U8 \ </ ^ -C-CO-NH - p ^ 0 ^ ^ - 'N \ y ^ ---- ^ H2OH (IV) ^ OCH8 COOR11, in which formula R ^ represents same as above, react with a carbamoylating agent such as R13-NCO (VIII) With an isocyanate of formula wherein R is a leaving group to give a carbamoyloxymethyl group in the 3-position, the nitrogen atom of which may be protected, followed by c) removing all carboxyl and N protecting groups if necessary, and finally d) isolating the desired compound of formula I whereby, if desired, the compound is previously converted into a non-toxic salt. Process according to Claim 1, characterized in that sodium (6R, 7R) -3-carbamoyloxymethyl-7-Z 2- (fur-2-yl) -2-methoxyiminoacetamido-7-ceph-3-em-U- carboxylate (syn isomer). Process according to Claim 2, characterized in that the sodium (6R, 7R) -3-carbamoyloxymethyl-7- [2- (fur-2-yl) -2-methoxyiminoacetamido] cef-3-em- The U-carboxylate (syn isomer) is obtained in Form II, or converted to Form II, the X-ray spectrum of which, using a Debye-Scherrer camera, radius 111 », 6 mm, and copper K ^ s 1.5 ^ + 18 A radiation, is as follows: : d I d I 8.78 60 3.1 »9 1U 7.81 9 3.07 6 (wide) 6.65 25 2.91 8 U, 68 100 (wide) 2.77 6 1+, 1 + 5 10 2.32 3 (wide) 1 », 20 10 2.19 2 3.76 20 (wide) 2.08 2 65623 29 A process for the preparation of antibiotics with active (6R, TR) -3-carbamoyloxymethyl-N- [2- (fur-2-yl) -2-methoxyiminoacetyl] uniclo7-cef-3-em-1-carboxylic acid in the form of HH \ -C-CO-NH - ^ S \ ^ 0 'iN A-CH2O-CO-NH (I) 0' 'and och3 C00H in the form of a syn-isomer or a mixture of syn- and anti- isomers having a content of 90% of the syn-isomer, the same as in the case of a gift, having the same meaning as in (a) a condenser with a formulation of HH l 1 S H2 ^ _ | --YN J ---- N ^ J --CHgO-CO-NHR (II) 0 [mu] m; oor11 11 12 color R is the same as the carboxyl group, and the R is the N-group, or is the addition of the carboxyl group; -C-C00H (III) \ / "'0- H ^ 0CH3 or b) to give a mixture with the formula