CS249523B2 - Method of cephalosporine's derivatives preparation - Google Patents

Description

The present invention relates to novel cephalosporin derivatives of formula I

wherein R ² is an unbranched or branched alkyl containing 1 to 4 carbon atoms, allyl, 2-butenyl or 3-ibutenyl or a group of the formula

R ³ —C — R ⁴

AND

COOH wherein R ³ and R ⁴ are each independently hydrogen, methyl or ethyl or R ³ and R ⁴ together with the carbon atom to which they are attached form a C 3 -C 5 cycloalkylidene ring and non-toxic pharmaceutically acceptable salts thereof, physiologically hydrolyzable esters and solvates thereof. The present invention also relates to a process for their preparation.

British Patent No. 1,399,086 contains general claims comprising a large number of cephalospoirins of the formula

wherein R is a hydrogen atom or an organic group, R ^a is an etherally bonded organic group bonded to an oxygen atom through a carbon atom, B is and P is an organic group. However, the 2-aminothiazol-4-yl group has not been identified as an R substituent and there is no mention that P can be N-methylpyrrolidiummethyl (or another fully saturated nitrogen containing cycle that is attached to the 3-methyl portion of the molecule via a nitrogen atom and containing another substituent on the nitrogen atom, U.S. Patents No. 3,971,778, 4,024,133, 4,024,137, 4,064,346, 4,033,950, 4,079,178, 4,091,209,

092 477 and 4 093 803 have similar claims.

U.S. Patent No. 4,278,793 discloses general claims comprising a significant number of cephalosporin derivatives of the general formula

wherein R 1, R 2, R 3, R 4, X and A include the general definitions of the corresponding substituents of the compounds of formula I. However, in the 20 paragraphs the definitions of various substituents, 78 pages of structural formulas and 225 examples do not claim that A may be N-methylpyrrolidinium methyl a fully saturated nitrogen containing cycle) which is attached to the 3-methyl portion of the molecule via a nitrogen atom and which contains an additional substituent on its nitrogen atom. British patent

No. 1,604,971 is consistent therewith and has substantially identical claims. British Patent Application Publication No. 2,028,305 A, although not formally related, contains the same broad claims, but mentions as a substituent A only a hydrogen atom in the examples.

West German DOS 2 805 655 claims 7- [2- (2-aminothiazol-4-yl) -2- (syn-methoxyiminoacetamido] cef allo spartanic acid derivatives of the general formula

wherein R ¹ NH is an optionally protected amino group, R ² is a halogen atom or an optionally substituted hydroxyl, thiol or amino group, and COOR is an optionally esterified carboxyl group. The patent claimed that if R ² is amino, may be disubstituted and the substituents together with the nitrogen atom can form inter alia a pyrrolidino group. However, there is no claim for an N-methylpyrrolidinium methyl group (or any other quaternary ammonium group) and the R ² substituent cannot be attached to the 3-position via a methylene group.

U.S. Pat. No. 4,278,671 claims 71 [2- (2-aminothiazol-4-yl) -2- (syn-methoxyimiinoacetamido) cephalosporin derivatives of the general formula

743523 wherein R 2 NH is optionally protected amino and R 3 is hydrogen or "nucleophilic compound residue". The term "nucleophilic compound residue" is broadly defined and it is then stated that R ³ "may alternatively be a quaternary ammonium group". As quaternary ammonium groups, pyridinium, variously substituted pyridinium, quinolinium, picolinium and lutidinium are claimed. There is no suggestion that a quaternary ammonium group may be a fully saturated, nitrogen-containing heterocyclic ring that is bonded through a nitrogen atom and which contains an additional substituent on its nitrogen atom. British Patent No. 1,581,954 is consistent therewith and has substantially identical claims. Other patents of the same inventors that are not formally related have similar claims and include U.S. Pat. Nos. 4,098,888, 4,020,899, 4,205,180 and 4,298,606 and British Patent No. 1,536,281.

U.S. Pat. No. 4,168,309 claims cephalosporin derivatives of the general formula

O i!

R ~ C ~ C ~ NH II

N

-i X

COO

Θ

CH

CH,

XJ '3

COOH wherein R is phenyl, thienyl or furyl, R ^a and R ^b are independently hydrogen, alkyl, cycloalkyl, phenyl, naphthyl, thienyl, furyl, carboxy, alkoxycarbonyl or cyano or R ^a and R ^b together with a carbon atom, to which they are attached form a cycloalkylidene or cycloalkenylidene ring, m and n are 0 or 1 such that the sum of m and n is 0 or 1, the substituents R 1, together with the nitrogen atom to which they are attached, are broadly defined but cannot be saturated five-membered ring. Example 5 is a compound of formula

British Patent No. 1,591,439 is analogous and has substantially identical claims. It is not mentioned in this patent that the R substituent may be 2-aminothiazol-4-yl or that the imino substituent does not contain a carboxyl group.

The present invention relates to cephalosporin derivatives of formula I

wherein R ² is a straight or branched (C 1 -C 4) alkyl, allyl, 2-hutenyl or 3-butenyl or group of the general formula

R ³ —C — R ⁴

AND

COOH where R ³ and R ⁴ are independently hydrogen, methyl or ethyl or R ³ and R ⁴ may form with the carbon atom to which they are attached a cycloalkylidene ring containing from 3 to 5 carbon atoms and non-toxic pharmaceutically acceptable salts and physiologically hydrolyzable esters. Also included within the scope of the present invention are the solvates (including hydrates) of the compound of formula I as well as the tautomeric forms of the compounds of formula I, for example the 2-iminothiazolidin-4-yl form of the 2-aminothiazol-4-yl substituent.

As can be seen from structural formula I, the compounds of formula I have a "son" or "Z" configuration relative to an alkoxyimino (or alkenyloxyimino) group or a carboxy-substituted alkoxyimino group. Since the compounds are geometric isomeirs, certain amounts of "anti" isomers may be present. The present invention includes compounds of formula I containing at least 90% of the "syn" isomers. Preferred compounds of formula I are "syn" isomers which are free of "anti" isomers.

Pharmaceutically acceptable salts of the compounds of formula I include inorganic bases such as alkali metal salts (e.g., sodium and potassium salts) and alkaline earth metal salts (e.g., calcium salts), ammonium salts, organic base salts (e.g., triethylamine, procaine, phenylethylbenzyl, amine, dibenzylethylenediamine and other organic bases used in the chemistry of penicillins and cephalosporin and salts with acids (for example, salts with hydrochloric, hydrobromic, formic, nitric, sulfuric, methanesulfonic, phosphoric, acetic or trifluoroacetic acids) Physiologically hydrolyzable esters include acyloxyalkyl esters such as (lower) alkanoyl (lower) alkyl esters such as acetoxymethyl, acetoxyethyl, pivoyloxymethyl and the like. Salts of esters and esters may be formed with any of the carboxyl groups of the compounds of formula I.

The compounds of formula I exhibit considerable anti-bacterial activity on various Gram-positive and Gram-negative bacteria and are useful for treating bacterial infections in animals, including humans. The compounds of formula (I) may be formulated for parenteral use, in a conventional manner using known pharmaceutical carriers and excipients, and may be administered in unit dosage form or in multi-dose containers. The compositions may be in the form of solutions, suspensions, or emulsions in oily or aqueous vehicles, and may contain conventional dispersing, suspending, or stabilizing agents. The compositions may also be in the form of a powder which is reconstituted prior to use, for example, sterile pyrogen-free water. The compounds of formula I may also be formulated as suppositories using conventional suppository compositions such as cocoa butter or other glycerides. If desired, the compounds of the invention may be administered in combination with other antibiotics such as penicillins or other cephalosporins.

In preparing unit dosage forms, the compositions preferably contain from about 50 to about 1,500 mg of active ingredient of Formula I. The dosage for treating adult humans is preferably from about 500 to about 5,000 mg per day, depending on frequency and mode of administration. When administered intramuscularly or intravenously to adult humans, a total dose ranging from about 750 to about 3000 mg per day in divided doses is sufficient, although higher doses of some compounds may be desirable in Pseudomonas infections.

Preferred compounds of formula I are those wherein R ² is methyl or ethyl or R ³ and R ⁴ are independently hydrogen or methyl. In the most preferred compounds, R ² is methyl or R ³ and R ⁴ are methyl. In the primary efficacy evaluation of the compounds of the invention, the minimum inhibitory concentrations (MICs) of the compounds and the two reference compounds (cefotaxime and ceftazidime) were determined by a two-fold step-by-step agar dilution in Mueller-Hinton agar for 32 test organism strains in six groups. The MIC geometric diameters determined for these tests are shown in Tables 1 and 4.

fe

OCH, (Cefotaxime; comparative compound)

AND

COOH (Ceftazidime; reference compound)

(test compounds)

All test compounds were found to be more active than cefotaxime in the (G -) - II and (G -) - III groups of the test organisms, with the most preferred compound Ia being significantly more active. All test compounds are more active than ceftazidime in the (G +) - Ia and (G +) - Ib groups of test organisms, with the most preferred compound Ia being significantly more active than ceftazidine in all groups of test organisms except (G-) -III, which is somewhat more sensitive to ceftazidim.

Absorption of the most preferred compound Ia and comparative compounds (cefotaxime and ceftazidime) was determined in mice after a single intramuscular injection of test compound (dissolved in 0.1 M phosphate buffer; pH 7) at a dose of 20 mg / kg. Blood samples were collected from orbital folds in heparinized capillary tubes and tested in Mueller-Hinton medium using Morganella Morgania A-9695 as test organism. Blood levels at various time intervals, half-life (t _1/2 ) and area under the curve (AUC) are shown in Table 2.

Tests have also been performed to identify organisms resistant to preferred compounds of formula Ia, cefotaxime and ceftazidime. The MIC concentrations of the three compounds per 240 Enterobacteriaceae strains were determined in Mueller-Hinton medium and an MIC value equal to or greater than 8 for at least one of the test compounds was determined as an indicator of the resistant organism. Of the 240 strains, 27 'were found to be resistant to at least one of the test compounds. The results showing 3 Compound Ia-resistant organisms, 15 Cefazidime-resistant organisms and 18 Cefotaxime-resistant organisms are shown in Table 3.

TABLE 1 MIC Geometric Diameter (µg / ml) G -) - Ia (G-) plb (6) (G-) - II (5) (G -) - III (6) Compound (G +) - Ia (5 strains) (G +) - Ib (5) Ia; R ² = methyl 1,2 3.1 0,025 0.13 0.33 2.8 Ib; R ² = ethyl, 1.4 3.1 0,087 0.32 1.0 5.6 Ic; R ^z = isopropyl 1.4 3.6 0.35 1.3 3.2 11 Id; R ² = allyl 1,8 3.6 0.53 1.1 2.4 13 Cefatoxim ' ³ ' 1.0 2.2 0.015 0.37 4.1 22nd Ceftazidim ' ³ ' 5.1 12 0,070 1.7 2.6 1,8 (G +) -Ia: Penicillin-sensitive S. (G +) - Ib: Penicilliri-resistant: (G—) -Ia: Cephalethin-sensitive 'aureus (5 strains) S. aureus (5 strains) N. coli (2 strains) Kl. pneumoniae (1 strain) and Pr.

mirahills (2 strains) (G-) -Ib: Cephalothin-resistant N. coll (3 strains) and Kl. pneumoniae (3 strains) [G-] - II: Pr. morganii (1 strain), Ent. cloacae (2 strains) and Ser. marcescens (2 strains) (G-) - III: Ps. aeruginosa (6 strains) (a) average of five experiments

TABLE 2

Blood levels after intramuscular administration to mice (20 mg / kg)

Compound Blood levels (jug / ml) t _1/2 , AUC minutes after application (minutes) (^ g / h / ml)

10 20 May 30 45 60 90 la; R ² = methyl ^(a) 20.7 19.6 13.6 8.8 4 ~ 0.9 17 13.5 Cefotaxime ^(b) 27.8 19.3 13 9.1 4.6 1,2 15 Dec 14.9 Ceftazidime ^(c) 21.5 18.4 14.9 8.7 4.4 ~ 0.8 17 13.8

(a) average of 2 tests (b) 1 test (c) average of 3 tests

TABLE 3

Resistance (MIC = 18 µg / ml) to one or more test compounds from 240 Enterobacteriaceae strains in Mueller-Hinton medium

Organism Geometric mean MIC (jug / ml) number of strains la Ceftazidim Cefotaxim

Escherichia coli 1 0.25 32 8 Escherichia coli 1 4 0.5 8 Klebsiella pneumoniae 1 2 16 0, Enterobacter aerogenes 3 0.25 32 13 Enterobacter aerogenes 1 4 8 32 Enterobacter cloacae 1 0.13 4 8 Enterobacter cloacae 3 0.5 40 50 Enterobacter cloacae 3 1.6 > 63 > 63 Enterobacter cloacae 1 > 32 > 63 > 63 Citrobacter freundii 2 0.35 45 32 Citrobacter species 1 0.03 > 63 32 Proteus vulgaris 1 0.06 8 8 Morganella morganii . 1 0.06 32 32 Serratia marcescens 1 1 1 16 Serratia marcescens 1 2 8 16 Serratia marcescens 2 2.8 2 11 Serratia marcescens 1 4 8 63 Serratia marcescens 1 8 16 8 Serratia marcescens 1 32 > 63 > 63 total number of resistant strains 27 Mar: 3 15 Dec 18

48323

COOH (test compound) TABLE 4

The geometric mean MIC (g g / ml)

Test organism Compound Ie

Compound

Cefotaxime ^(a) Ceftazidime ^{(a |} (G +) - Ia (5 strains) 14 1.0 5.1 (G +) - Ib (5) 33 2.2 12 (G-) -Ia (5) 0.066 0.015 0.070 ( G-) -Ib (6) 0.79 0.35 1.7 (G-) -II (5) 1.2 4.1 2.6 (G-) -III (6) 4.0 22 1, 8 (G +) - Ia: Penicillin-sensitive S. aureus (5 strains) (Gl -) - Ib: Penicillin-resistant S. aureus (5 strains) (G-) -Ia: Cephalothin-sensitive E. coli (2 strains) ), Kl pneumoniae (1 strain) and P. mirabilis (2 strains) (G -) - Ib: Cephalothin-resistant E. coli (3 strains) and Kl pneumoniae (3 strains) (G -) - II: Pr morgani (1 strain), Ent. cloacae (2 strains) and Ser. Marcescens (2 strains) (G-) -III: Ps. seruginosa (6 strains) (a) average of five experiments.

Compound Ie appears to be more active than cefotaxime on the (G-) -II test organisms and significantly more active than cefotaxime on the (G.) - ΠΙ test organism (Ps. Aeruginosa). It was more active than ceftazidime on all groups of gram-negative test organisms with the exception of (G -) - III (Ps. Aeruginosa), which are somewhat more sensitive to ceftazidime.

It is an object of the present invention to provide a process for preparing compounds of formula I. There are two basic processes for converting readily available starting cephalosporins to other cephalosporins having different substituents at the 7 and 3 positions. Alternatively, the desired 3-position suhstituent may be introduced first, and then the 7-position substituent may be substituted, and the compounds of Formula I may be prepared by any of the methods, but it is preferable to first introduce the desired suhstituent. The preferred procedure is shown below in Reaction Scheme 1, while the alternative procedure is shown in Reaction Scheme 2. The abbreviations "Tr" refer to the trityl (triphenylmethyl) group, which is the preferred group for aminoisk protection full. The abbreviation "Ph" stands for phenyl. Thus, part of the —CH (Ph) 2 molecule is a benzhydryl group that is a preferred carboxyl protecting group. Reaction Schemes 3 and 4 below show the preparation of compound Ie wherein R ³ and R ⁴ are methyl.

249523 Reaction Scheme 1

C

II

N

C00C _from H ₅

OH (10

• c-COO li II N

TrHN / 7

OR (IV)

TrHN

(IN M)

Ν

2. deprotection

(I)

Reaction Scheme 2

P hCH.CQNH — ^p >

O

-Ν '

CbjOy

COOCH (Ph)

PCI _with pyridine

COOCH (Ph)

Na.1

PhCHgCONH0

CHU

C.

COOCH (Ph) _z h ₃ c-n

PhCH2 CONH

deacylation (xn v

(xm

TrHN

H ^ N

(XII i)

249323 Reaction Scheme 3

C-COOf- /

COOCH1PhL

OF

HjC CgHj

COOCICH ^ (II 1 <a. ') (V)

TrHN

TrHN

1. Dicyclohexylcarbodiimide (DCC)

2. (V)

C

N

1. Bis (trimethylsilyl) acetamide (BSA)

2. (IIIa ‘) + PCls

· - {- ^:!

Λ a) h ₃ ccch ₃ coocch ^

íVís.)

TrHN

C

II

N

O I

WjCCCW ₃

CONH

COOCHIPh)

OF.

COOCtCH2 v

C-CONH il N deprotection ^x o

I h, ccch ₅ • 3 i ³

COOH

Ύ CH _; 'Θ coo ^

CH f <2.)

Reaction Scheme 4

PhCH ^ CONH —li '

_T OH CH

COOCH (Ph) ₂

PC4 _S pyrbd. Lh p hCH ^ CONH

(I) (In Her) (IX)

329323

PhCH ^ CONH-

CH 2 I COOCH (Ph), (X)

Ph CH 2 CONH

N

-AND

Θ

N

fxn ^H of ^N

4 4 · 4 · N l —´N \ /

ThHN S

(OCHtPh). d.42.A.cylation>

'CH,

COOCH (Ph), (IIa1 DCC

CONH h $ ccch ₂

CH,

CH,

CH (XII)

COOCHfPht (Vila.) ΙΘ

CH,

Deprotection

Ν - _χ zs c

II

N • conh

H _k N o

H? CCCH ?.

COOH

O'

T '% ~ ^rj coo f la

CH,

While the above reaction schemes show preferred multistep processes for preparing compounds of Formula I, it is understood that other starting materials and processes can be used to prepare intermediates used in the key steps of each reaction scheme. Thus, a key step in Reaction Scheme 1 is the reaction of a compound of Formula VII with N-methylpyrrolidine. Compound VII may itself be prepared by other methods. Similarly, the key step in Reaction Scheme 2 is the acylation of the compound

XII with compound IV. Both compounds XII and IV can be prepared by other methods.

The key step in Reaction Scheme 3 is the reaction of VIa with N-methylpyrrolidine. Via can itself be prepared by other methods. Similarly, the key step in Reaction Scheme 4 is the acylation of compound XII with compound IIa ‘. Both compound XII and IIIa * can be prepared by other methods.

The present invention provides a process for the preparation of compounds of formula (I)

wherein R ² is straight or branched (C 1 -C 4) alkyl, allyl, 2-butenyl or 3-butenyl or a group of formula R ₃ -C 4

COOH wherein R ³ and R ⁴ are independently hydrogen, methyl or ethyl or R ³ and R ⁴ together with the carbon atom to which they are attached may form a cycloalkylidene ring containing from 3 to 5 carbon atoms and netoxiokých pharmaceutically acceptable salts, physiologically hydrolyzable esters and solvates, characterized in that a compound of formula XIV is reacted

B * HN

II

C-c -

Ί

(XIV), with N-methylpyrrolidine to give a compound of formula (XV) wherein R ² is as defined above, B ¹ is a conventional carboxyl protecting group and B ² is a conventional amino-protecting group

after which all protecting groups are removed by conventional methods or the compound of formula XIVa is reacted

O

cooe ³ wherein R ³ and R ⁴ are as defined above, B ¹ and B ³ are common carboxyl protecting groups and B ² is a common amino protecting group, with N-methylpyrrolidine to form a compound of formula XVa

^CH 3 is then removed in a conventional manner by all protecting groups.

The reaction is carried out in a non-aqueous organic solvent such as methylene chloride, chloroform, ethyl ether, hexane, ethyl acetate, tetrahydrofuran, acetonitrile, or the like, or mixtures of these solvents. The reaction is conveniently carried out at a temperature of from about -10 ° C to about + 50 ° C, preferably at room temperature. At least one mole of N-methylpyrrolidine per mole of compound XIV or XIVa is used for the reaction, preferably from about 50% to 100% excess of N-methylpyrrolidine is used.

Carboxyl protecting group, which are suitable py use as substitueinty B ¹ and B ³ in the above reaction, are well known and include arylalkyl groups such as benzyl, ρ-methoxybenzyl, p-nitrobenzyl and diphenylmethyl (henzhydryl J alkyls such as tert-butyl, haloalkyl, such as 2,2,2-trichloroethyl, and other carboxyl protecting groups described in the literature, for example, in British Patent No. 1,399,086. Preferably, carboxyl protecting groups are used which are readily de-acidified. Particularly preferred carboxyl protecting groups are benzhydryl and tert-butyl groups.

Amino-protecting groups which are suitable for use as substituents of B ² are also known in the literature and include trityl-and acyl groups such as chloroacetyl. Amine protecting groups that are readily removed by acid, such as trityl, are preferred.

Preparation 1

Λ

WITH'

COOC ^ Hs

OR ^Z (lil)

Ethyl (Z) -2-methoxyimino-2- (2-tritylaminothiazol-4-yl) Jacetate (Ha)

A mixture of ethyl 1- (Z) -2-hydroxyimino-2- (2-tritylamino-tbiazol-4-yl), acetate (II) (5.00 grams, 10.9 mmol), CH 3 I (2.04 mL, 32.8 mmol) and KCOCO ((4.54 g, 32.8 mmol) in anhydrous dimethylsulfoxide (100 mL) was stirred at room temperature overnight and then poured into water (250 mL). The solid was washed with water and dried to give the title compound (5, 15 g, quantitative yield), mp 115 ° C (dec.).

NMR: With CDCl3, ppm:

1.32 (3 H, t),

3.98 (3 H, s),

4.30 (2 H, q),

6.42 (1 H, S),

7.2 (1 H, m),

7.25 (15H, s).

Compounds IIIb, IIIc and IIId were prepared by the general procedure above, but substituting methyl iodide for the corresponding iodide.

Compound R ² Yield (%) Mp (° C) References' ⁰ tt (° Cj lila methyl 100 ALIGN! 115 (dec) about 120 (dec.) Illb ethyl 67 97 to 98 * IIIc isopropyl 26 51 to 55 * Illd allyl * *

The ester was hydrolyzed without isolation <Tetrahedron, 34, 2233 (1978).

C · ~~ CO H>!

rx - -, i?

(Z) -2-Methoxyimino-2- (2-tritylamino-thiazol-4-yl) -acetic acid (IVa)

The ethyl ester IIIa prepared according to Preparation No. 1 (6.00 g, 12.7 mmol) in ethanol (120 mL) was treated with 2N NaOH (12.7 mL) at room temperature overnight. The reaction mixture was adjusted to pH 8 by addition of dry ice powder and the solvent was evaporated under reduced pressure. The residue was dissolved in water (100 mL) and the solution was acidified to pH 2 with 1N hydrochloric acid and then extracted with ethyl acetate (3 x 50 mL). The combined extracts were washed with saturated aqueous sodium chloride solution, dried and evaporated. The residue was crystallized from ethyl acetate / hexane to give 5.56 g (98% yield) of the title compound. Mp 138-143 ° C (dec.).

NMR: δ CDCl3, ppm:

3.89 (3 H, s),

6.52 (1 H, s),

7.2 (15H, s).

Compounds IVb, IVc and IVd are prepared by the general procedure outlined above.

Compound R ² Yield (%) mp (° C, dec.) Literature ^'0

t. t. (° Ό, dec.)

IVa methyl 98 138 to 143 about 140 IVb ethyl 85 140 to 145 neuvedeno IVc isopropyl 85 166 to 169 about 170 Illd allyl 66 170 to 178 about 170

⁽¹⁾ Tetrahedron, 34, 2233 (1978).

Preparation 3

Benzhydryl-3-hydroxymethyl-7-phenylacetamido-3-cephem-4-carboxylate (VIII)

To a stirred suspension of phosphate buffer (pH 7, 162.5 mL) and wheat bran (20 g, dry), 7-phenylacetamidocephalosporanic acid sodium salt (5 g, 12.1 mmol) was added in one portion at room temperature. The progress of the reaction was monitored by high pressure liquid chromatography until complete hydrolysis (5 hours). The suspension is then filtered, the bran separated and the filtrate cooled to 5-10 ° C for extractive esterification. To the cooled solution was added methylene chloride (32 mL) followed by a 0.5 M solution of diphenyldiazomethane in methylene chloride (24 mL). The pH was then adjusted to 3.0 by addition of 28% phosphoric acid and allowed to warm to 20 ° C after one hour. Heptane (56 mL) was added slowly and the resulting crystalline title compound was isolated by filtration to give the title compound 3.0 g (50%).

Preparation 4

To a suspension of PCl 3 (8.3 g, 40 mmol) in dichloromethane (100 mL) was added pyridine (3.2 g, 40 mmol) and the mixture was stirred at 20 ° C for 20 min. hydroxymethyl 7-phenylacetamido-3-cephem-4-carboxylate, prepared according to Preparation 3 (5.1 g, 10 mmol), at -40 ^° C. The mixture is stirred at -10 ° C for 15 minutes and then The mixture was allowed to stand for 7 hours at -10 ° C to -15 ° C. To the cooled solution (-20 ° C) was added propane-1-diol (10 mL), and the mixture was allowed to stand at -20 ° C for 16 hours. The solution was washed with ice water (2 x 20 mL), saturated aqueous sodium chloride (10 mL), dried (MgSO 4) and concentrated in vacuo. CHCl 3 and n-hexane (2: 1) and chromatographed on a silica gel column (200 g) using the same solvent as the eluent. The title compound was evaporated in vacuo and the residue was triturated in n-hexane to give the title compound (2.1 g, 51%), mp> 110 ° C (dec.

ID: in KBr

400, 2 800, 1 785, 1 725 cm ¹ .

v. / 1 max

265 nm (eI 160).

NMR: OPP ^m m ^s ° ^'d "+ CDCb: 3.69 (2H, sj,

4.43 (2H, sj,

5.09 (1H, d, J = 4.5Hz),

5.24 (1H, d, J = 4.5Hz),

6.87 (1H, sj,

7.3 (10 H, m).

7.3 (10, i.

Example 1

7 - [(Z-2-methoxyimino-2- (2-aminothiazol-4-yl) acetamido) -3 - ((1-methyl-1-pyrrolidinium) methyl] -3-cephem-4-carboxylate (Ia)

A. Benzhydryl-3-chloromethyl-7 - [(Z) -2-imethoxyxyimino-2- (2-tritylaminothiazol-4-yl) acetamido] -3-cephem-4-carboxylate (Via ¹ )

Benzhydryl-7-amino-3-chloromethyl-3-cephem-4-carboxylate prepared as described in Preparation 4 (2.29 g, 5.52 mmol) in CH 3 CN (57 mL was treated with bis (trimethylsilyl) acetamide (BSA, 4,09 ml,

16.6 mmol) at 50 minutes to give a clear solution. To this solution was added an acid chloride solution prepared from (Z) -2-methoxyimino-2- (2-tritylaminothiazol-4-yl) acetic acid (IVa) (2.04 g, 4.60 mmol) and PCl 2. (1.15 g, 5.52 mmol) in methylene chloride (20 mL). The mixture was stirred at room temperature for 30 minutes, poured into cold water (200 mL) and extracted with ethyl acetate (3 x 100 mL). The combined extracts were washed with brine, dried and evaporated. The remaining syrup (4 g) was chromatographed on a silica gel column (150 g) and eluted sequentially with 10: 1 and 3: 1 toluene: ethyl acetate to give the fractions containing the desired compound, evaporating to give 2.61 g (68 °). (o) Compound VIa in the form of an amorphous powder.

NMR: δ CDCl3, ppm:

3.50 (2H, sj,

4.02 (3H, s,

4.33 (2H, sj,

4.98 (1 H, d)

5.87 (1 H, q),

6.65 (1H, sj,

6.90 (1 H, s),

7.3 (25 H, m).

B. Benzhydryl 3-iodomethyl-7 - [(Z-2-methoxyimino-2- (2-tritylaminothiazol-4-yl) acetamido] -3-cephem-4-carboxylate (VIIa‘j)

A mixture of the 3-chloromethyl derivative (Flag (1.50 g, 1.79 mmol) and NaI (1.34 g, 8.93 mmol) in methyl ethyl ketone (30 mL) was stirred at room temperature for one hour. in ethyl acetate (100 ml) and washed with water, aqueous Na2SO4 solution and brine. Drying and evaporation gave compound VIIa (1.47 g, 89 percent) as an amorphous powder.

243523

NMR: δ CDCL ·, ppm:

3.55 (2H, ABq);

4. OP (3H, s)

4.25 (2 H, s),

4.97 (1 H, d),

5.80 (1 H, q),

6.65 (1 H, s),

6.90 (1 H, s),

7.3 (25 H, m).

C. 7 - [(Z) -2-methoxyimino-2- (2-aminothiazol-4-yl) acetamido) -3 - ((1-methyl-1-ipyrrolidinium) methyl) -3-cephem-4-carboxylate (la)

A mixture of VIIa ¹ (4.5 g, 4.83 mmol) and N-methylpyrrolidine (0.65 mL, 6.28 mmol) in CH 2 Cl 2 (45 mL) was stirred at room temperature for 20 min. The quaternary salt of protected cephalosporin was collected by filtration and treated with 90% trifluoroacetic acid (40 mL) for one hour at room temperature, then the mixture was evaporated under reduced pressure at a temperature below 20 ^° C. ether to give trifluoroacetic acid salt of compound 1a (2.40 g), which is dissolved in methanol (5 mL) and treated with a 1 M solution of 2-ethylhexisic acid sodium salt in ethyl acetate (8 mL) at room temperature for After addition of ethyl acetate (100 ml), the precipitate (1.94 g) was filtered off, and the crude product was found to be 7% and contained a 1: 8 A ³ and A ² isomer in high pressure liquid chromatography. liquid chromatography was repeated three times times (Lichrosorb RP-17, 8 xx 300 mm), eluting with 50% aqueous methanol or 0.01 M ammonium phosphate buffer (pH 7.2) containing 5% methanol to give 35 mg (1.5 percent) of the title compound in the form of a colorless powder. Purity found (HPLC) 90%. Mp 150 ° C (dec.).

ID: rX cm ^-1 :

770, 1660, 1620.

_T tt7> phosphate buffer pH? , _Ί

UV: λ _{I, x} nm (ε):

235 (16,200), 258 (15,400) ..

NMR:? ^D3 ° ppm:

2.31 (4 H, m),

3.08 (3H, s);

3.63 (4 H, m),

4.09 (3 H, s),

5.43 (1H, d, J = 4.8Hz),

5.93 (1 H, d),

7.08 (1 H, s).

Example 2

7 - [(Z) -2-methoxyimino-2- (2-aminothiazol-4-yl) acetamido) -3 - ((1-methyl-1-pyrrolidinium) methyl] -3-cephem-4-carboxylate (1a) )

To a stirred solution of 20.4 g (21.9 mmol) of Vila ¹ in 150 ml of anhydrous methylene chloride was added 2.42 g (28.5 mmol) of 1-methylpyrrolidine at room temperature in one portion. The mixture was stirred for 5 minutes and then poured into 1000 ml of ether with vigorous stirring. The resulting precipitate was filtered off, washed with ether (5 x 30 ml) and dried in vacuo. 19.3 g of protected product are obtained in the form of a pale yellow powder.

τ Λ ΚΒΓ _ 1

IC: in _lliax cm d

400, 1780 (s), 1740, 1075, 1530.

Thin layer chromatography: ethanol-CHCL · (1: 3), Rf = 0.30 (Rf = 0.95 for VIl‘a).

The solid was dissolved in 185 mL of trifluoroacetic acid / water (99: 1), stirred at room temperature for one hour, and concentrated to about 30 mL at less than 10 ° C. The concentrate was poured into 1000 ml of ether with vigorous stirring and the resulting precipitate was filtered off, washed with ether (5 x 40 ml) and dried in vacuo. 10.6 g of a pale yellow powder are obtained. The powder was dissolved in 20 mL of methanol and the solution was filtered. To the filtrate was added 45 mL of 0.8 M ethylhexanoic acid sodium salt in ethyl acetate. The resulting suspension was poured over 400 mL of ethyl acetate and filtered to give 8.08 g of a solid which is a mixture of the title compound and the corresponding ^{2 2} -isomer ( ³ 3/2 ² = 1: 8) (according to high pressure liquid). chromatography, Lichrosorb RP-18 10- to 15% methanol in 0.01 M phosphate buffer, pH 7). A second batch of 28.9 g (31.0 mmol) of VIIa 'yielded 16.0 g of crude product (A ³ / A ² = 1: 8). Isolation of the desired AMsomer from the combined crude products (24.08 g) using preparative high pressure liquid chromatography (System 500, Watera Associates, Prep PAK 500 / C18, 5- to 10% CH 3 OH) gave 769 mg of 1a.

Example 3

7 - [(Z) -2-methoxyimino-2- [2- (2-aminothiazol / 4-yl) acetamido] -3 - [(1-methyl-1-pyrrolidinium) methyl] -3-cephem- 4-carboxylate (1a)

A series of experiments were performed to determine the effect of the solvent, the amount of solvent and the reaction time on the yield of compound Ia and on the ratio A ³ / A ² in the reaction product. The general procedure was as follows.

To a suspension of 3-iodomethyl Villa ¹ (45 mg, 0.048 mmol) in said amount in the solvent was added a solution of N-methylpyrrolidine (0.01 mL, 0.087 mmol) in ether (0.1 mL) and the mixture was stirred at room temperature for that period. The reaction mixture was diluted with ether (5 mL) and the precipitate formed was filtered off and treated with 90% trifluoroacetic acid. The mixture was stirred for one hour and then evaporated to dryness under reduced pressure below 20 ° C. The ratio of Δ ³ / Δ ² in the product obtained is determined by high pressure kaipaline chromatography (Lichrosorb RP-18, mobile phase, 0.01 Mammonium phosphate buffer (pH 7.2 containing 15% CH ³ OH, retention time Δ ³ = 6.60 minutes, Δ ²⁾ 5.56 minutes) The product yield and the ratio of ³ 3/2 ² isomers for each ip is shown below.

Try C. Solvent Ratio Vila (g) to solvent (ml) Reaction time (min) Yield (%) Ratio A ³ / A ² 1 CH2Cl2 1: 20 15 Dec 73 1/8 2 CH2Cl2-ether (1/10) 1: 100 15 Dec 25 4/1 3 ethyl acetate-ether (1/10) 1: 100 15 Dec 27 Mar: 4/1 4 ethyl acetate-ether (1/10) 1: 100 60 64 2/1 5 ether 1: 100 15 Dec 31 6/1 6 ether 1: 100 60 62 3/1 7 ether 1: 60 15 Dec 55 3,5 / 1 8 ether 1: 60 60 82 1/1 say 4 B. Diphenylmethyl-7 - [(Z) -2-ethoxyimino-2-

7- [Z (2-ethoxyimino-2- (2-aminothiazol-4-yl) acetamido] -3 - [(1-methyl-1-pyrrolidinium) methyl] -3-cephem-4-carboxylate (1b)

A. Benzhydryl-3-chloromethyl-7 - [(Z) -2-ethoxyimino-2- (2-tritylaminothiazol-4-yl) acetamido] -3-cephem-4-carboxylate (VIb)

To a solution of (Z) -2-ethoxyimino-2- (2-tritylaminothiazol-4-yl) acetic acid (IVbj (1.095 g, 2.4 mol) in dichloromethane (20 mL) was added phosphorus pentachloride (500 mg). After stirring for 1 hour at room temperature, the mixture was added all at once to an ice-cooled solution of compound V (1.083 g, 2.4 mmol) and his (trimethylsilyl) acetamide (1 mL) in dichloromethane (20 mL). % aqueous sodium bicarbonate solution (200 mL), extracted with chloroform (100 mL), washed with water, dried (MgSO 4) and evaporated under reduced pressure.

The residue is chromatographed on a silica gel column. Elution with chloroform gave Compound VIb as an amorphous powder 1.76 g (86%).

NMR: With CDCl3, ppm:

1.40 (3H, t, CH 2 CH 3),

3.53 (28H, ABq, 2-CH2);

4.37 (2H, a, —CH 2 Cl),

4.60 (2H, q, - CH 2 CH 3),

4.90 (lH, d, 6-H),

5.89 (1H, d, 7-H);

6.88 (1H, s);

6.88 (1H, s, thiazole-H),

6.91 (1H, s, benzhydryl-CH).

- (2-tritylaminothi azole-4-yl jacetamido] -3-iodomethyl-3-cephem-4-carboxylic ^l boxyláit (VIIb)

A mixture of VIb (1.07 g, 1.25 mmol and NaI (562 mg, 2.75 mmol) in acetone (20 mL) was stirred for one hour, filtered and the filtrate was poured into water and extracted with ethyl acetate. washed successively with 5% aqueous Na 2 S 2 O 3 solution, water and saturated aqueous sodium chloride solution, dried over magnesium sulfate and evaporated to give 1.04 g (89%) of VIIb.

NMR: ^Γ1κ ' ^ΐ! ppm:

3.55 (2H, q, 2-CH2);

4.27 (2H, s, CH 2 -I),

5.02 (1H, d, 6-H);

5.87 (1H, d, 7-H);

6.68 (1H, s, thiazole-H),

6.93 (1H, s, benzhydryl-CH).

C. 7 - {[Z] -2-Ethoxyimino-2- (2-aminothiazol-4-yl) acetamido} -3 - ((1-methyl-1-pyrrolidinium) methyl) -3-cephem-4 -carboxylate (1b)

A mixture of VIIb (333 mg, 0.35 mmol) and N-methylpyrrolidine (60 mg, 0.7 mmol in CH 2 Cl 2 5 mL) was stirred at room temperature for 0.5 h and then evaporated in vacuo. The residue was washed with ether and dissolved in 90% trifluoroacetic acid. After standing at room temperature for 0.5 hour, the mixture was concentrated under reduced pressure. Ether was added to the concentrate to separate the quaternized product, which was then filtered and dissolved in a small amount of methanol. The solution was chromatographed on an HP-2 column (40 mL). Elution with 30% aqueous methanol followed by 25052 lyophilization gave 0.062 g of a mixture of Δ ² and Δ ³ isomer (Δ ² : Δ ³ = 5: 1). The mixture was purified by HPLC (Lichrosorb RP-18, 8 x 300 mm, 15% methanol) and the desired Δ ³ isomer (Ib) was isolated as a light yellow iprášku, 4.9 mg (2.7%).

UV: λ ^{výp leaching I, H7} nm (ε):

235 (15,000), 258 (14,000).

NMR: .delta ° ^IL2 ppm:

1.43 (3 H, t),

2.33 (4H, m).

3.10 (3 H, S);

3.64 (4 H, m),

4.36 (2 H, q),

5.44 (1 H, -d),

5.95 (1 H, d),

7.08 (1 H, s).

Example 5

7 - [(Z) -2- (2-propoxyimino) -2- (2-aminothiazol-4-yl) acetamido] -3 - [(1-methyl-1-pyrrolidinium) methyl] -3-cephem-4- carboxylate (Ic)

A. Diphenylmethyl-3-chloromethyl-7 - [(Z) -2- (2-ipropoxyimino) -2- (2-tritylaminothiazol-4-yl) acetamido] -3-cephem-4-carboxylate (More)

A mixture of (Z) -2- (2-propoxyimino) -2- (2-trityarnaminothiazol-4-yl) acetic acid (IVc) (707 mg, 1.5 mmol) and phosphorus pentachloride (344 mg, 1, 65 mmol) in dichloromethane (14 mL) was stirred at room temperature for one hour and then poured into a solution of compound V (677 mg, 1.5 mmol) and bis (trimethylsilyl) acetamide (1.1 mL, 4.5 mmol) in The reaction mixture was stirred at room temperature for 30 minutes, diluted with ethyl acetate (200 mL) and water (3 x 100 mL), dried over sodium sulfate and evaporated to give 1.4 g (100%) of the title compound. ).

IC: in max cm:

360, 3,020, 3,060, 2,960, 1,785, 1,725,

680, 1,520, 1,500, 1,450, 1,375, 1,300,

250, 1,160, 1,160, 1,090, 1,010, 990,

840, 740, 700.

UV: λ nm (ε):

240 (24,600), 260 (20,700).

NMR: δ ^{CDCl 3} ppm:

1.35 (6H, d, I = 6Hz)

3.50 (2 H, s),

4.35 (2 H, s),

4.58 (1 H, m, J = 6 Hz),

5.00 (1H, d, J = 4.5Hz),

5.91 (1H, d-d, J = 4.5, 9 Hz, d, D 2 O,

J = 4.5 Hz)

6.68 (1 H, s),

6.88 (1 H, s),

7.25 (25 H, s).

B. Diphenylmethyl-3-iodomethyl 1-7 - ((Z) -2- (2-propoxyimino) -2- (2-tritylaminothiazole-4-acetamido) -3-cephem-4-carboxylate (Villa)

A mixture of VIc (500 mg, 0.55 mmol) and sodium iodide (248 mg, 1.66 mmol) in acetone (10 mL) was stirred at room temperature for 50 min. After evaporation, the residue was dissolved in ethyl acetate (15 mL), washed sequentially with 10% aqueous sodium thiosulfate solution (10 mL), water (10 mL) and brine (10 mL), dried over sodium sulfate and evaporated to give the title compound.

494 mg (90%) of the title (Vilo). compounds listed IR: Umax cm ^-1 : 3,360, 3,040, 3,020, 1,680, 1,600, 1,520, 1,300, 1,230, 1,150, 900, 840, 750, 700. 2,960, 1,500, 1115, 1,785, 1,450, 1,080, 1 720, 1 370, 990, UV: Λn'x ^H nm (ε): 240 (24,900), 260 (19,400). NMR: δ ^{r: DC k} ppm: 1.30 (6H, d, J = 6 Hz),

3.37 and 3.70 (1 H each, d, J = 16 Hz),

4.22 (2 H, s),

4.55 (1 H, m, J = 6 Hz),

4.95 (1H, d, J = 4.5Hz),

5.83 (1H, d-d, J = 4.5 and 9 Hz, d D2O), 6.66 (1H, s),

6.87 (1 H, s),

7.25 (25 H, s).

C. 7- [(Z) -2- (2-Propoxyimino) -2- (2-aminothiazol-4-yl) acetamido] - 3 - ((1-methyl-1-pyrrolidinium) methyl) -3-cephem -4-carboxylate (Ic)

A mixture of Vile (545 mg, 0.55 mmol) and 1-methylpyrrolidine (70 mg, 0.82 mmol) in dichloromethane (10 mL) was stirred at room temperature for 30 min and diluted with ether (100 mL). The precipitate formed. filtered. A solution of the precipitate in 90% trifluoroacetic acid (4.5 mL) was stirred at room temperature for 30 minutes, then evaporated in vacuo. The residue was triturated with ether to give 317 mg of crude product, which was chromatographed on an HP-20 column (50 mL), eluting with a mixture of water (500 mL) and 30% CH 3 OH (500 mL). The eluate with 30% methanol is concentrated and lyophilized. This gives 109 mg of a mixture of Δ ² and Δ ³ isomers (Δ ² / Δ ³ = 6: 1). 100 mg of this mixture was purified by high pressure liquid chromatography (Lichrosorb RP-18, 15%).

MeOH to give 5 mg (3%) of the desired compound Ic.

UV: λ £ ^H x ^{7 buffer} nm ( _ε ) _:

236 (15,100), 252 (14,600).

NMR: ^{11) 20} ppm:

1.42 (6H, d, J = 6Hz),

2.33 (4H, s);

3.10 (3 H, s),

3.65 (4H, s);

3.83 and 4.23 (1H each, d, J = 17 Hz),

5.45 (1 H, d, J = 4.5 Hz),

5.95 (1 H, d, J = 4.5 Hz),

7.05 (1 H, s).

Example 6

7 - [(Z) -2-allyloxyimino-2- (2-aminothiazol-4-yl) acetamido] -3 - [(1-methyl-1-pyrrolidinium) methyl] -3-cephem-4-carboxylate (Id )

A. Benzhydryl-7 - [(Z) -2-allyloxyimino-2- (2-tritylaminophthiazol-4-yl) acetamido] -3-chloromethyl-3-cephem-4-carboxylate (Vid)

To a suspension of Compound V (1.35 g, 3 mmol) in methylene chloride (20 mL) was added bis- (trimethylsilyl) acetamide (1.1 mL, 4.5 mmol) and the mixture was stirred at room temperature for 30 min. to obtain a clear solution. A mixture of (Z) -2-allyloxyimino-2- (2-tritylaminothiazol-4-yl) acetic acid (IVd) (1.40 g, 3.0 mmol) and phosphorus pentachloride (690 mg,

3.3 mmol) in methylene chloride (20 ml) was stirred at room temperature for 15 minutes and poured into a solution of trimethylsilylated compound V all at once. The mixture was stirred at room temperature for 20 minutes and diluted with ethyl acetate (200 ml), washed with aqueous bicarbonate solution. sodium and water, dried and evaporated under reduced pressure. The oily residue was purified by silica gel column chromatography (Wako-gel, C-200, 30 g). The column was eluted with chloroform and the fractions containing the desired product were combined. yield 2.32 g (89%), mp 100-115 ° C (dec.).

IC: in max cm *

990, 1,790, 1,730, 1,680, 1,530, 1,380,

250, 1160, 1020.

NMR: δ ^rDCl5 ppm:

3.50 (2 H, 2-H),

4.32 (2H, 3-CH 2),

4.6 to 6.1 (7H, m, CH 2 CH = CH 2 and 6.7-H),

6.70 (1H, s, thiazole-1H),

6.90 (1H, s, PI 12 CH),

7.1 to 7.6 (30H, m, phenyl protons). Analysis: for C48H40N5O5S2Cl. 1/3 CHCl3: calculated:

64.05 ° C, 4.45% H, 7.73% N, 7.08% S, 7.82% Cl, found:

63.13, 63.99% C, 4.61, 4.64% H,

7.50, 7.30% N, 6.85, 6.95% S,

7.55, 7.46% Cl.

B. Benzhydryl-7 - [(Z) -2-allyloxyimino-2- (tritylaminothiazol-4-yl) acetamido] -3-iodomethyl-3-cephem-4-carboxylate (Vild)

A mixture of Vid (2.30 g, 2.65 mmol) and sodium iodide (2 g, 131.3 mmol) in acetone (15 mL) was stirred at room temperature for one hour and then evaporated under reduced pressure. A solution of the oily residue in ethyl acetate (200 mL) was washed with 10% sodium thiosulfate and water, evaporated under reduced pressure to give Vild as an amorphous powder, which was used in the next step without further purification. Yield 2.52 g (99%).

C. 7- [(Z) -2-Allyloxyimino-2- (2-aminothiazol-4-yl) acetamido] -3 - [(1-methyl-1-pyrrolidinium) methyl] -3-cephem-4-carboxylate (Id)

A mixture of Vild (478 mg, 0.5 mmol) and N-methylpyrrolidine (0.05 mL, 0.5 mmol) in methylene chloride (5 mL) was stirred at room temperature for 20 min, diluted with ether (50 mL). thus precipitating the quaternized product (yield 500 mg). The mixture of quaternized product and trifluoroacetic acid (2 ml) was allowed to stand at room temperature for 1.5 hours, diluted with ether to precipitate the crude trifluoroacetic acid product salt (yield 265 mg) which was chromatographed on an HP-20 column. (1.8 x 18 cm). The column was eluted with water and 30% aqueous methanol. The methanolic eluate was evaporated under reduced pressure and the residue was lyophilized to give an amorphous powder (yield 124 mg) containing the desired product (17%) and the corresponding A ^{from the} isomer (83%). The mixture was purified by high pressure liquid chromatography (Lichrosorb RP 18; 0.01 Μ NH 4 H 2 PO 4 (pH 7): CH 3 OH = 85: 15). The eluate was acidified to pH 3 with dilute hydrochloric acid and chromatographed on an HP-20 column (1.8 x 10 cm). The column was eluted with water and then 30% aqueous methanol. The methanolic eluate was evaporated under reduced pressure and the residue was lyophilized. The title compound (Id) was obtained as an amorphous powder (yield 13 mg, 5.1%). Mp 155 ° C (dec.).

THREE KBr —1

IC: in max cm h

600—2 800, 1,770, 1,670, 1,610, 1,530,

200

UV: λ SV ^puIr nm (ε):

235 (16,600), 253 (15,600).

NMR: δ ^1.30 ppm:

2.1 to 2.5 (4H, m, pyrrolidine-H),

3.10 (3H, s, _NCH3);

3.4-3.8 (4H, m, pyrrolidine-H),

5.95 (1H, d, 4Hz, —H),

7.10 (1H, s, thiazole-1H);

Example 7

7- (2- (2-aminothiazol-4-yl) - (Z) -2- (2-carboxyprop-2-oxyimino) acetamido) -3 - [(1-methyl-1-pyrrolidinium) methyl-3] -cephem-4-carboxylate (Ie j

A. Benzhydryl-3-chloromethyl-7 - [(Z) -2- (2- t -butoxycarbonylprop-2-oxyimino-2- (2-tritylaminothiazol-4-yl) acetamido] -3-cephem-4- carboxylate (Va)

Procedure 1

A mixture of (Z'j -2- (2-tert-butoxycarbonylprop-2-oxyimino) -2- (2-tritylaminothiazol-4-yl) acetic acid (IIIa)) (1.94 g, 3.6 mmol), DDC (742 mg, 3.6 mmol) and N-hydroxybenzotriazole (486 mg, 3.6 mmol) in tetrahydrofuran (45 mL) was stirred at room temperature for 45 min while dicyclohexylurea was precipitated. The mixture was stirred overnight at room temperature and then evaporated in vacuo. The residual oil was dissolved in chloroform (20 mL), washed with saturated aqueous sodium bicarbonate solution and saturated aqueous sodium bicarbonate. The residue was dissolved in n-hexane / chloroform (1: 2) and passed through a column of silica gel (40 g) in the same solvent mixture. The title compound was evaporated in vacuo to give 1.3 g (39%) of compound Va, mp > 100 ° C (decomposition).

ID: 9 cmX cm ^-1 :

990, 1790, 1715, 1690.

UV: λ λ nm:

240 (E160 280), 265 (E160 190).

NMR: δ ^{1 / κ} · ¹³ ppm:

1.45 (9H, s);

1.63 and 1.66 (6H, s each),

3.49 (2H, broad s),

4.34 (2 H, s),

4.96 (1 H, d, J = 4.5 Hz),

5.90 (1H, d-d, J = 4.5 and 7.5),

6.66 (1 H, s),

6.86 (1 H, s),

7.0 to 7.5 (25H, m)

8.23 (1H, d, J = 7.5Hz).

Procedure 2

A solution of compound V (1.86 g, 4.49 mmol) in CH ₃ CN (46.5 mL) was treated with bis (trimethylsilyl) acetamide (3.33 mL, 13.5 mmol) at room temperature for 50 min. minutes and a clear solution was obtained. To this solution was added the acid chloride prepared from compound IIIa ¹ (2.56 g, 4.49 mmol) and phosphorus pentachloride (1.12 g, 5.38 mmol) in methylene chloride (26 mL). The mixture was stirred at room temperature for 30 minutes, poured into cold water (100 mL) and extracted with ethyl acetate (3 x 50 mL). The combined extracts were washed with brine, dried and evaporated. The evaporated syrup (5 g) is chromatographed on a silica gel column (100 g) eluting with a 10: 1 mixture of toluene and ethyl acetate. Fractions containing the desired compound were combined and evaporated to give 2.84 g (65%) of Compound Va.

B. Benzhydryl-7- [(Z) -2- (2-tert-butoxycarbonylprop-1-oxyimino) -2- (2-tritylaminothiazol-4-yl) acetamido] -3-iodomethyl-3-cephem-4 -carboxylate (Via)

A mixture of Va (500 mg, 0.53 mmol) and NaI (240 mg, 1.6 mmol) in acetone (3 mL) was stirred at room temperature for 2 h and then evaporated in vacuo to the residue. The organic phase was washed with 10% sodium thiosulfate (5 ml) and brine (5 ml), dried over magnesium sulfate and evaporated in vacuo to give 540 mg (90%) of VIa as amorphous powder, m.p. 106 ^DEG C. (decomposition).

IC: V max cm h

350, 1790, 1690.

UV: λ mX nm 240 (λX 270), 265 (λX 190).

NMR: δ ¹ “ ¹ ppm ppm:

1.44 (9H, s);

1.65 (6H, s);

3.54 (1 H),

4.28 (2 H, s),

4.98 (1H, d, J = 4.5Hz),

5.85 (1H, d-d, J = 4.5 and 7.5 Hz),

6.70 (1 H, s),

6.90 (1 H, s),

7.1 to 7.5 (25H, m).

C. Z- (2- (2-Aminothiazol-4-yl) - (Z) -2- (2-carboxyprop-2-oxyimino) acetamido) -3 - [(1-methyl-1-pyrrolidinium) methyl] - 3-cef-4-carboxylate (Ie)

A mixture of iodomethyl derivative Via (538 mg, 0.51 mmol) and N-methylpyrrolidine (0.079 mL, 0.076 mmol) in CH 2 Cl 2 (10.8 mL) was allowed to stand at room temperature for 30 minutes and then diluted with ether (80 mL). The resulting precipitate was filtered off and washed with ether. 420 mg of quaternized product are obtained, from which the protecting group is removed by treatment with 90% trifluoroacetic acid (4.2 ml) for 1 hour at room temperature. The reaction mixture was then evaporated to dryness. The crude salt of compound Ia with trifluoroacetic acid (245 mg, quantitative yield) is obtained, which is a 1: 4 Δ ³ and Δ ² isomer mixture. The crude product was purified by high pressure liquid chromatography [Lichrosorb RP-18, 4 x 300 mm, eluting with 0.01 M ammonium phosphate buffer (pH 7.0) containing 10% CH 3 OH]. The fractions containing the desired product were combined and evaporated to a small volume. The concentrate was adjusted to a pH of about 2 by the addition of 1 M hydrochloric acid and passed through an HP-20 column (2 x 15 centimeters) to remove inorganic salts. The column was washed with water (1000 mL) and eluted with 30% methanol. The eluate was evaporated and lyophilized to give 21 mg (10%) of the title compound (Ie) as a colorless powder. Melting point 160 ° C (dec.).

ID: Χ cm ^-1 :

400, 1775, 1610.

UV: λ ^ ^{1ονγραΐΓρΗ7} η _Μ (ε):

237 (15,700), 257 (155,500).

NMR: ^{λ) 2} ° ppm:

1.65 (6H, s);

2.3 (4H, m)

3.09 (3 H, s),

3.6 (4 H, m),

4.0 (2H, m);

5.44 (1H, d, J = 4.8Hz),

5.94 (1 H, d),

7.15 (1 H, s).

Example 8

The general procedure of Example 7 was followed except that (Z) -2- (2-tert-butoxycarbonylprop-2-oxyimino) -2- (2-tritylaminothiazol-4-yl) acetic acid was replaced by an equimolar amount. (Z) -2- (tert-butoxycarbonylmethoxyimino) -2- (2-tritylaminothiazol-4-yl) acetic acid, (Z) -2- (1-tert-butoxycarbonylethoxyimino) -2- (2-tritylaminothiazol-4) (Z) -2- (2-tert-butoxycarbonylbut-2-oxyimino) -2- (2-tritylaminothiazol-4-yl) acetic acid, (Z) -2- (3-tert-yl) acetic acid; butoxycarbonylpent-3-oxyimino) -1- (2-tritylaminothiazol-4-yl) acetic acid, (Z) -2- (1-tert-butoxycarbonylcycloprop-1-oxyimino) -2- (2-tritylaminothiazol-4-yl) (acetic acid), (Z) -2- (1-tert-butoxycarbonylcyclobut-2-oxyimino) -2- (2-tritylaminothiazol-4-yl) acetic acid and (Z) -2- (1-tert-butyloxycarbonylcyclopent) 1-oxyimino) -2- (2-tritylaminothiazol-1-yl) acetic acid to give

7- (2- (2-aminothiazol-4-yl) - (Z) -2- (carboxymethoxyimino) acetamido) -3 - [(1-methyl-1-pyrrolidinium) methyl] -3-cephem-4-carboxylate,

7- (2- (2-aminothiazol-4-yl) - (Z) -2- (1-carboxyethoxyimino) acetamido] -3 - [(1-methyl-1-pyrrolidinium) methyl] -3-cephem-4 -carboxylate,

7- [2- (2-aminothiazol-4-yl) - (Z) -2-carboxybut-2-oxyimino) acetamido) -3 - ((1-methyl-1-pyrrolidinium) methyl) -3-cef em-4-carboxylate,

7- (2- (2-aminothiazol-4-yl) - (Z) -2- (3-carboxypent-3-oxyimino) acetamido) -3 - ((1-methyl-1-pyrrolidinium) methyl) -3- cefem-4-carboxylate,

7- (2- (2-aminothiazol-4-yl) - (Z) -2- (1-carboxycycloprop-1-oxyimino) acetamido] -3 - [(1-methyl-1-pyrrolidinium) methyl] -3-cephem -4-carboxylate,

7- (2- (2-aminothiazol-4-yl) - (Z) -2- (1-carboxycyclobut-1-oxyimino) acetamido] -3 - [(1-methyl-1-pyrrolidinium) methyl] -3- cefem-4-carboxylate a

7- (2- (2-aminothiazol-4-yl) - (Z) -2- (1-carboxycyclopent-1-oxyimino) acetamido) -3 - ((1-methyl-1-pyrrolidinium) methyl) -3- cef em-4-carboxylate.

Claims (3)

A process for the preparation of cephalosporin derivatives of the general formula I (I) wherein R ² is a straight or branched alkyl having 1 to 4 carbon atoms, allyl, 2-butenyl or 3-butenyl or a group of the formula R ³ -C 'R' ¹ COOH wherein R ³ and R ⁴ are independently hydrogen, methyl or- ethyl or R ³ and R ⁴ together with the carbon atom to which they are attached may form a cycloalkylidene ring containing from 3 to 5 carbon atoms, and nontoxic pharmaceutically acceptable salts , physiologically hydrolyzable esters and solvates thereof, characterized in that the compound of formula XIV or XIVa NO. 1 NcII M. O II -C OR ² · O II cNH NH0 R ³ -C- ^Rk COOBu ³ / ~ CGOB X, NI- ^CH 7 CH.I iL CGOB (XI V) wherein R ² , R ³ and R ⁴ are as defined above, B ¹ and B ³ are common carboxyl protecting groups, B ² is a common amino protecting group, reacted with N-methylpyrrolidine to form a compound of formula VX or XVa 8 ² HN ZJ cH N O II CNH _n J ¹ RC O R I, COO0 ³ hCOGB ΙΘ Θ / X ch, - ni ¹ CH, (X V a), then all protecting groups are removed in a conventional manner, whereupon the compound of formula I is optionally converted into non-toxic pharmaceutically acceptable salts, physiologically hydrolyzable esters or solvates. 2. The process of item 1 for preparing 7- [2- (2-aminothiazol-4-yl) - (Z) -2- (2-carboxypropyl-2-oxyiminoacetamido) -3 - [(1-methyl-1- pyrrolidinium) methyl] -3-cep-4-carboxylate or a non-toxic pharmaceutically acceptable salt, physiologically hydrolyzable ester or solvate, characterized in that benzhydryl-7 - [(Z) -2- (2-tert. butoxycarbonylprop-2-oxyimino) -2- (2-tritylaminothiazol-4-yl) acetamido] -3-iodomethyl-3-cephem-4-carboxylate with N-methylpyrrolidine in an organic solvent, followed by deprotection in the obtained intermediate and or optionally converting the compound obtained into non-toxic pharmaceutically acceptable salts, physiologically hydrolyzable esters or solvates. 3. A process according to claim 1 wherein R &lt; ^{2 &gt;} is methyl, ethyl, isopropyl or allyl or a non-toxic pharmaceutically acceptable salt or solvate thereof, wherein the starting 3-iodomethyl derivative is reacted with N-methylpyrrolidine in an organic solvent. or a mixture of organic solvents to give the corresponding pyrrolidinium derivative and finally, by cleavage of the protecting groups from the pyrrolidinium derivative, 7 - [(Z) -2-methoxyimino-2- (2-aminothiazol-4-yl) acetamido] -3 - [(1-imethyl- 1-pyrrolidinium) methyl] -3-cephem-4-carboxylate or 7 - [(Z) -2-ethoxyimino-2- (2-aminothiazol-4-yl) acetamido) -3 - ((1-methyl-1) -pyrrolidinium] methyl] -3-cephem-4-carboxylate or 7 - [(Z) -2- (2-propoxyimino) -2- (2-aminothiazol-4-yl) acetamido] -3 - [(1-methyl) -1-pyrrolidimum) methyl] -3-cephem-4-carboxylate or 7 - [(Z) -2-allyloxyimino-2- (2-aminothiazol-4-yl) acetamido] -3 - [(1-methyl-1-pyrrolidinium) methyl] -3-cephem-4-carboxylate and / or optionally converting these unprotected derivatives into non-toxic pharmaceutically acceptable salts or solvates.