CS249512B2 - Method of cephalosporine's derivatives preparatio - Google Patents

Abstract

Novel cephalosporin derivatives of the formula <IMAGE> wherein R<1> is hydrogen or a conventional amino-protecting group, and R<2> is a straight or branched alkyl group containing from 1 to 4 carbon atoms, allyl, 2- butenyl or 3-butenyl, or is a group <IMAGE> wherein R<3> and R<4> each are independently hydrogen, methyl or ethyl, or R<3> and R<4>, taken together with the carbon atom to which they are attached, may be a cycloalkylidene ring containing from 3 to 5 carbon atoms, and nontoxic pharmaceutically acceptable salts, physiologically hydrolyzable esters and solvates thereof exhibit potent antibacterial activity and can be incorporated into pharmaceutical compositions.

Description

The present invention relates to novel cephalosporin derivatives of formula I

0) where

R ² is straight or branched alkyl · 1 to 4 carbon atoms, allyl,

2-butenyl or 3-butenyl or a group of the formula

R ³ —C — R 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 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 cephalosporins of the formula

COOH where R is a hydrogen atom or an organic group, R ^a is an ether-bonded organic group bonded to an oxygen atom through a carbon atom, V is as an R substituent and there is no mention that P may be N-methylpyrrolidiummethyl which is attached to the 3-methyl moiety of the molecule through a nitrogen atom and which contains an additional substituent on the nitrogen atom US Patent Nos. 3,917,178, 4,024,133, 4,024,137, 4,064,346, 4,033,950, 4,079178, 4,091209 ,

Nos. 092 477 and 4,093,803 have similar claims to the rinine derivatives of the general formula

U.S. Patent No. 4,278,793 discloses general claims involving a significant number of cephalosporin.

The substituents R1, R2, R3, Rd, X and A include the general definitions of the corresponding substituents of the compounds of formula I. However, in the 20 paragraphs of the various substituent definitions, 78 pages of structural formulas and 225 examples, there is no 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. 6,604,971 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-aminothlazol-4-yl) -2- (syn) methoxyiminoacetamido] -cephalosporanic acid derivatives of the general formula

S or S - O

Z and P are an organic group. However, the 2-aminothiazol-4-yl group has not been identified

wherein R 1 H 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 the methylene group.

U.S. Patent No. 4,278,671 claims 7- [2- (2-aminothiazol-4-yl) -2- (syn) methoxyiminoacetamido] cephalosporin derivatives of the general formula

wherein R 2 NH is an optionally protected amino group and R 3 is hydrogen or a "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 pyridiinium, 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. 1,581,854 is in accordance with it and has substantially identical claims. Other patents of the same inventors that are not formally related have similar claims and include U.S. Patent Nos. 4,098,888, 4,203,899, 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

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 the 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; which, together with the nitrogen atom to which they are attached, are broadly defined, but cannot be, inter alia, a saturated five-membered ring, as an example of 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-aminoithlazol-4-yl or that the iunino substituent does not contain a carboxyl group.

The present invention relates to a cephalosporin derivative of the formula I

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

R3-C-R4

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 compounds of formula I as well as the tautomenic forms of the compounds of formula I, for example the 2-iminothiazolin-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 "syn" or "Z" configuration relative to an alkoxyimino (or alkenyloxyimino) or carboxy-substituted alkoxyimino group. Since the compounds are geometric isomers, certain amounts of "anti" isomers may be present. The present invention includes compounds of formula I containing at least 90% of the "syin" 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 base salts 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, phenylethylbenzylamine, dibenzylethylenediamine and other organic bases used in the chemistry of penicillins and cephalosporins) and acid salts (for example, salts with hydrochloric, hydrobromic, formic, nitric, sulfuric, methanesulfo249512 new, phosphoric, acetic, or trifluoroacetic acid) and other used in the chemistry of penicillins and cephaosporins. Physiologically hydrolyzable esters include acyloxyalkyl esters, for example (lower) alkanoyl (lower) alkyl esters such as acetoxymethyl, acetoxyethyl, pivaloyloxymethyl, and the like.

The compounds of formula 1 exhibit considerable antibacterial activity against various Gram positive and 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 discrete, suspending or stabilizing agents. The compositions may also be in the form of a dry 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. The compounds of the invention may if desired be administered in combination with other ^aatib: orc such as penicillins or other cephalosporins.

In preparing unit dosage forms, the compositions preferably contain from about 50 to about 1500 mg of active ingredient of Formula I. The dosage for treating adult humans is preferably from about 500 to about 3000 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 Pseudomoinas 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 evaluation of the effect 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 two-step serial agar dilution in Mueller-Hinton agar for 32 strains of test organisms in six groups. these tests are shown in Tables 1 and 4.

N

C-CONH

R?

In Q 'v

(Cefotaxime; reference compound)

cooh (Ceftazidim; comparative compound)

C ....... _c QK / jr 'Z \ _л o (') (test compounds)

All test compounds were found to be more active than cefotaxime in the (G -) - II and (G -) - III groups of 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 ceftazidime in all groups of test organisms except (G -) - III, which is somewhat more sensitive to ceftazidime.

Absorption of the most preferred compound Ia and comparative compounds (cefotaxime and ceftazidime) was determined in mice after a single imtramuscular 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

Morganii A-9695 as a 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 carried out to identify organisms resistant to the preferred compounds of formula Ia, cefotaxime and ceftazidime. The MIC concentrations of these three compounds per 240 Enterobacteriaceae strains were determined in Mueller-Hlnton's medium and the MIC value of the same or greater than 8 for at least one of the tested 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 Ceftazidime-resistant organisms, and 18 Cefotaxime-resistant organisms are shown in Table 3.

TABLE 1

Compound

MIC (/ g / ml) [G - +] - Ia (G +) - Ib (G -) - Ia (G -) - Ib (G -) - II (G -) - III (5 kme- ( 5) (5) (6) (5) (6)

Ia; R ² = methyl

Ib; R ² = ethyl,

Ic; R 2 = isopropyl

Id; R2 = allyl Cefatoxim (a) Ceftazidime ^(a)

1,2 3.1 0,025 0.13 0.33 2.8 1.4 3.1 0,087 0.32 1.0 5.6 1.4 3.6 0.35 1.3 3.2 11 1,8 3.6 0.53 1.1 2.4 13 1.0 2.2 0.015 0.35 4.1 22nd 5.1 12 0,070 1.7 2.6 1,8

(G +) -Ia: Penicillin-sensitive S. aureus (5 strains) (G +) - Ib: Penicillin-resistant S. aureus (5 strains) (G -) - Ia: Ciephaloohin-sensitive E.-coli (2 strains) ), Kl. pneumoniae (1 strain) and Pr. mirabilis (2 strains) (G-) -I'b: Cephalo-thin-resistant E. coli (3 strains -a 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

Ί 'i? 1 * 7

TABLE 2

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

Compound Blood levels (^ g / ml) t 1/2 AUG minutes after application (minutes) ^ (g / h / ml)

10 20 May 30 45 60 90 la; R 2 = methyl ^) 20.7 19.6 13.6 8.8 4 ~ 0.9 17 13.5 Cefotaxime ^ ¹ 27.8 19.3 13 9.1 4.6 1,2 15 Dec 14.9 Ceftazidim ^) 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 = ≥8 / (g / ml) to one or more test compounds from 240 strains

Enterohacteriaceae in Mueller-Hinton medium

Organism Number of strains Geometric mean MIC (µg / ml)) la Ceftazidim Cefotaxim

Escherichia coli 1 0.25 32 8 Escherichia coli 1 4 0.5 8 Klebsiella pneumoniae 1 2 16 0.13 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 27 Mar: 3 15 Dec 18

strains of rh © Λ.

^{, N} Y ^{X -} CH, 7W 1

I Θ ý '\ J

COO /

CI4 ₃

NgH

i) "Tn"" ^coNH \ ^c

H ₃ CC-CH ₃

COOH (test compound)

TABLE 4

Geometric MIC (gg / ml)

Test organism Compound Ie

Compound

Cefotaxime ( ^a )

Ceftazidim ^)

(G +) - Ia (5 ikmen) 14 1.0 5.1 (G +) - Ib 33 2.2 12 (G-) - Ia (5) 0,066 0.015 0,070 (G-) - Ib (0) 0.79 0.35 1.7 (G-) - II (5) 1,2 4.1 2, G (G -) - in (G) 4.0 22nd 1,8

(G +) - Ia: Penicillm-sensitive S. aureus (5 strains) (G +) - Ib: Penicililn-ressstentm S. aureus (5 strains) (G -) - Ia: E. coli (2 strains), Kl. pneumoniae (1 strain), and Pr. mirabii-s (2 strains) (G—) -Ib: Ceplhalothin-resistant E. coli (3 strains) and K1. pneumoniae (3-strains) (G -) - II: Ex. morgani (1 strain), Ent. cloacae (2 strains and Ser. Marcescens (2 strains)

(G - -) - 111: Ps. aeruginosa (6 strains) (a) average of five experiments.

It can be seen that compound Ie is more active than cefotaxime on the test organisms of the group (G-J-II and significantly more active than cetotaxime on the test organism (Ps. Aeruginosa) from the (G -) - III group. Gram negative 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 others - cephalosporins having different substituents at the 7 and 3 positions. Alternatively, the desired substituent can be introduced first - at the 3-position and then the substituent at the 7-position is exchanged. Compounds of formula I - can be prepared however, it is preferable to introduce the desired substituent at the 7-position and then introduce the desired substituent at the 3-position. The preferred procedure is shown below in Reaction Scheme 1, while the alternative procedure is shown in Reaction Scheme 2. The abbreviations "TI" mean trity! a (triphenylmethyl) group which is a preferred amino protecting group. The abbreviation 'Ph' stands for phenysupine. Thus, part of the —CH (Ph) 2 molecule is a benzhydryl group, which is the preferred carboxyl protecting group. Reaction Schemes 3 and 4 below show the preparation of Compound Ie wherein R 3 and R ⁴ are methyl.

241512

Reaction Scheme 1

Tt-HN

C-COQH ti ^X OR ^Z (IV)

J 2 CH ₂ Ct

COOCH (

J. (V) where c ~~ COMH ₀ CH ₂ Cl ² 0R ⁴ COOCH (PK) ₂

CONH

COOCH (Pfz) ^ ( ^vl 4

2. deprotection

Reaction Scheme 2

PfbCH.CONH-i - (Ί

From X / ^ - cHaOM cooch (РРл)?

P (^ 1 _£ v

^ and C ^ CONH

COOOCrfPP ^ z.

(IX)

IN

СООСН (Ph)% ž-0 ψ

PhCH2 CONH

соосн (ры) ₂

СН ₃ (XI) Declaration

Г

СООСН (Pb) ž

P ' ^v}

(X //)

(χιιι)

Ý

G'

(I) »2W — Γ-γ ^ 'Ί

C00CH (Pb.) Z

Reaction Scheme 3

N - r - C - COOH

IIл и

TrHN N

^Ό и з сСси з СП0С (СН3) з IIIa) (V)

1. Dicyclohexylcarbodiimide (DCC)

2. (V) / 1. bis (trimethylsilyl) acetamide (BSA) 2. (IIIa ‘) + PC15

Fourty hCpu)% l (Va)

Na I

TrHN _gg

N · —τ— C ——COMH Г I

Λ ₀ > ~ ^-i ' ^Y ^ CH ₂ .

COOC H (РЬ-Ц

СООС (СМ ₃ ) з ií 'Wa' (Wa)

H ₃ ^C- O

Дсн ₂ -0

COOCH (PM2)

XjC ^—conH

Market ^S \ θ ¹ н _э сссн ₃

COOC (CH3) ₃ (VH3).

deprotection

Reaction Scheme 4

PhcH _{2 and} N ^H _{3 - (} C 3 ^{H 3} OH)

COOCH (Ph) '

PCs Pyramid

VΡΗΟΗΟΟΝΗ (Vili)

NaC

IX (IX)

ΡΙ-ια-ίθΟΝΗ ——

0 ^ ~

CH ₂ l

COOCH (Ph.) ₂ ^H ₃ CV °

⁽ X)

PhCH _? CO ^NH -Ί Θ Z _of ^N s _<M ^ _CH _®

COOCH2 (III)

... .. CH3 ⁽ XI) deacylation of ₃ (хи)

N — t — С - CONH й о ₃ сссн ₃ соос (сн ₃ ) ₃

СООСН (Ph.) 2 \ сн ₃ (external)

C — CONH II

N ^from deprotection

V <r.

h ₃ ccch ₃

COOH

While the above reaction ischimates show preferred multistep processes for the preparation of 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 acylation of compound XII with compound IIIa ‘. Both compounds XII and IIIa ‘can be prepared by other methods.

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

Ν — rΗχΝ (i)

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

R ³ -C ⁴ R ⁴ COOH, methyl or ethyl or R ³ and R ⁴ together with the carbon atom to which they are attached may form a cycloalkylldene ring containing from 3 to 5 carbon atoms and inetoxic pharmaceutically acceptable salts, physiologically hydrolyzable esters and solvates, characterized in that the compound of formula XVI is acylated wherein R ³ and R ⁴ are independently of each other a hydrogen atom.

(XV) or an N-silyl derivative thereof, wherein B ¹ is a hydrogen atom or a common carboxyl protecting group, an acid derivative of an acid of formula XVII wherein B ² is a common amino protecting group and R ² is as defined above and an optionally present carboxyl group in R ² is protected to form a compound of formula XV

N-C-COOH. Л л и o к / ₈ · ΗΝ \ _or 2.

(xv //)

(XV) wherein Β ¹ , B ² - and R 2 are as defined above, after which all protecting groups are removed by conventional means, whereupon the compound of formula (I) is optionally converted into non-toxic pharmaceutically acceptable salts, physiologically hydrolyzable esters or solvates.

Acylating derivatives of the acid of formula XVII include acid halides (and especially acid chlorides), mixed acid anhydrides (such as acid anhydrides formed with pivalic acid or haloformates such as ethyl chloroformate) and activated esters [such as esters that may be formed with N-hydroxybenztriazolene in the presence of a condensing agent such as dicyclohexylcarbodiinide). The acylation can also be carried out using the free acid of formula XVII and in the presence of a condensing agent such as dicyclohexylcarbodiimide, carbonyldiimidazole or an isoxazolium salt. As used herein, the term "acylating derivative" of an acid of formula XVII includes the free acids themselves in the presence of a condensing agent as set forth above. A preferred acid acylation derivative of formula XVII is an acid chloride, preferably used in the presence of an acid binding agent (and in particular a tertiary amine such as triethylamine, dimethylaniline or pyridine).

When the acylation is carried out with an acid halide, it is possible to use an aqueous reaction medium, but preferably a non-aqueous medium is used. If acid anhydrides, activated esters or free acid in the presence of a condensing agent are used for acylation, then the reaction medium should be inverse. Particularly preferred solvents for the acylation reactions are halogenated hydrocarbons such as methylene chloride and chloroform; tertiary amides such as dimethylacetamide or dmethylformamide as well as other conventional solvents such as tetrahydrofuran, acetonitrile and the like can also be used.

The acylation reaction can be carried out at a temperature of about -50 ° C to about +50 ° C. However, the reaction is preferably carried out at or below room temperature, most preferably at about -30 ° C to about 0 ° C. Preferably, the compound of formula XVI is commonly acylated using about a stoichiometric amount of the acylating agent of formula XVII, although a small excess (i.e., about 5-25%) of the acylating agent may also be used.

It is preferred that the compound of formula XVI is acylated in the form of an N-silyl derivative (when a non-free reaction medium is used). This reaction is preferably carried out in situ simply by adding a suitable silalating agent (for example, 3-methylsilylacetamide) to a solution of the compound of formula XVI before adding the acylating agent of formula XVII. Preferably, about 3 moles of silylating agent are used per mole of compound of formula XVI, although this is not critical. The silyl compound is readily cleaved after acylation by addition of water.

The acylated acids of formula XVII, including derivatives thereof with a carboxy-protected or amino-protected group, are known or can be prepared by known methods. Thus, (Z) -2- (2-t-butoxycarbonylprop-2-oxyimino) -2- (2-tritylamino-thiazol-4-yl) acetic acid is prepared according to the general procedure described in US Patent 4,258,041 and British Patent The melting points reported in these patents were 152-156 ° C (dec.), but the compound prepared by us had a melting point of 174-175 ° C (dec.).

P a v v l

Ethyl (Z) -2-methoxyimino-2- (2-tritylaminothiazol-4-yl) acetate (IIIa)

A mixture of ethyl (Z) -2-hydroxylmino-2- (2-tritylamino-thiazol-4-yl) acetate (II) (5.90 g, 10.9 mmol), methyl iodide (2.04 mL, 32.8 mmol) and potassium carbonate (4.54 g, 32.8 mmol) in anhydrous dimethylsulfoxide (100 mL) was stirred at room temperature overnight and then poured into water (250 mL), and the resulting precipitate was filtered off, washed with water. Drying gave the title compound (5.15 g, quantitative yield), mp 115 ° C (dec.).

NMR: <5 ^{CDCl 3} ppm

1.32 (3 H, t),

3, P8 (3H, 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 ² R

Yield tt (° C) literature ¹¹ (%) tt (° C)

lila methyl 100 ALIGN! 115 (dec) about 120 (dec.) Illb ethyl 67 97—98 * IIIc isopropyl 26 51—55 * Illd allyl * * *

The ester was hydrolyzed without isolation. ¹¹ Tetrahednon, 34, 2233 (1978).

C ont 2 (?

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.).

(Z) -2-Ni (dhoxyimino-2- (2-tritylaminothiazole, 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.

NMR: 5 ^cr) ci3ppm

3.89 (3 H, s),

6.52 (1H, s),

7.2 (15H, s).

Compounds IVb, IVc and IVd were prepared by the general procedure described above.

compound R ² yield (%) t. t. (° C, dec.) literature ¹ * tt (° C dec.) IVa methyl 98 138—143 about 140 IVb ethyl 85 140—145 neuvedeno IVc isopropyl 85 166—169 about 170 Illd allyl 66 170—178 about 170

^υ Tetrahedron, 34, 2233 (1978).

P rin g 3

Benzhydryl 3-hydroxymethyl-7-phenyl-Cetamido-3-cephem-4-carboxylate (VIII)

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

Preparation 4

Benzhydryl 7-amino-3-chloromethyl-3-cephem-4-carboxylate

To a suspension of PC15 (8.3 g, 40 mmol) in dl249512 chloromethane (100 mL) was added pyridine (3.2 g, 40 mmol) and the mixture was stirred at 20 ° C for 20 min. To the mixture was added benzhydryl 3-hydroxymethyl-7-phenylacetamido-3-cephem-4-carboxylate prepared as described in Preparation 3 (5.1 g, 10 mmol) at -40 ^° C with stirring. at -10 ^° C for 15 minutes and then allowed to stand at -10 ° C to -15 ° C for 7 hours. To the cooled solution (-20 ° C) was added pro-propain-1,3-diol (10 mL) and the mixture was allowed to stand at -20 ° C for 16 hours and then stirred at room temperature for 20 minutes. The resulting solution was washed with ice water (2 x 20 mL), saturated aqueous sodium chloride solution (10 mL), dried (MgSO 4) and concentrated in vacuo. The gummy residue (12 g) was dissolved in a mixture of CHCl3 and n-hexane (2: 1) and chromatographed on a silica gel column (200 g) using the same solvent as the eluent. The fractions containing the title compound were evaporated in vacuo and the residue was triturated in n-hexane to give the title compound (2.1 g, 51%) mp> 110 ° C (dec.).

IR: KBr 3400, 2700, 1785, 1725 cm -1. UV: λ * H 265 nm (E)% 160).

NMR δ D ^MS O-d 6 + CDCl 3

LY1H \. и pp, _m

3.69 (2 H, s),

4.43 (2H, s), 5.09 (1H, d, J = 4.5Hz),

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

6.87 (1 H, s),

7.3 (10, m).

Example 1

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

A. Benzhydryl-3-chloromethyl-7 - [(Z-2-methoxyimino-2- (2-tritylaminothiazol-4-yl) Jacetamido] -3-cephem-4-carboxylate (VIa ‘)

Benzhydryl-3-chloro-3-chloromethyl-3-cephem-4-carboxylate prepared according to 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 room temperature for 50 minutes to give a clear solution, to which is added an acid chloride solution prepared from (Z) 12-methoxyimino-12- (2-tritylaminothiazole-4-). yl) acetic acid (IVa) (2.04 g, 4.60 mmol) and PCl 3 (1.15 g, 5.52 mmol) in methylene chloride (20 mL) and stirred at room temperature for 30 min. 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. 10: 1 and 3: 1 toluene / ethyl acetate were sequentially combined and the fractions containing the desired compound were combined, evaporated to give 2.61 g (68%) of Via ¹ as amorphous. about powder.

NMR: .delta

3.50 (2 H, s),

4.02 (3 H, s),

4.33 (2 H, s),

4.98 (1 H, d),

5.87 (1 H, q),

6.65 (1 H, s),

6.90 (1 H, s),

7.3 (25 H, m).

B. Benzhydryl 3l) Methyl 7 - [(Z-2-methoxyethoxy-2- (2-tritylammothiazoll-4-yl) acetamido] -3-cephem-4-carboxylate (VIIa‘J)

Mixture of 3-chloro-methyl derivative (VIa ‘) (1.50 g,

1.79 mmol) and NaI (1.34 g, 8.93 mmol) in methyl ethyl ketone (30 mL) were stirred at room temperature for one hour. After evaporation of the solvent, the residue was dissolved in ethyl acetate (100 ml) and washed with water, aqueous sodium chloride solution and brine, and dried and evaporated to give compound VIIa (1.47 g, 89%) as an amorphous powder.

NMR: δ ^CDGh ppm

3.55 (2H, ABq);

4.00 (3 H, 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-21- (2-amino-thiazol-4-yl Jacetamido) -3 - ((1-methyl-11-pyrrolidinium) methyl) -3-cephem-4-carboxylate (fa)

A mixture of VIIa1 (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 minutes. Ether (300 ml) was added and the quaternary salt of protected cephalosporins was separated, which was filtered off and treated with 90% trifluoroacetic acid (40 ml) for one hour at room temperature. The mixture is then evaporated under reduced pressure at a temperature below 20 ° C. The residue was triturated with ether to give the salt of compound 1a with trifluoroacetic acid (2.40 g), which was dissolved in methanol [5 mL] and treated with a 1M solution of sodium 2-ethylhexanoic acid in ethyl acetate (8 mL). at room temperature for 30 minutes. After addition of ethyl acetate (100 mL), the precipitate (1.94 g) was filtered off. The crude product was found to be 7% by HPLC and contained a ratio of 1: 8 Δ ³ and Λ ² isomer. Purification of the product by high pressure liquid chromatography was repeated three times (Lichrosorb RP-18, 8x300 mm, eluting with 5% aqueous methanol or 0.01 M ammonium phosphate buffer (pH 7.2) containing 5% methanol to give 35 mg (1.5%). The title compound was obtained in the form of a colorless powder, and was found to be (high pressure liquid chromatography) 90%, mp 150 ° C (dec.).

IR: cm ^-1 1770, 1660, 1 620th

UV: A ^ ir ^{ltový buffer 'рИ 7} nm (ε)

235 (16,200),

258 (15,400).

NMR:

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 (1H, d),

7.08 (1H, s).

Example 2

7- [(Z) -2-Melhoxyimino-2- (2-amino-thiazol-4-yl) -amido] -3 - ((1-methyl-1-pyrrolidinium) methyl) -3-cephem-4-carboxylate (la)

To a stirred solution of 20.4 g (21.9 mmol) of Vila ¹ in 150 ml of anhydrous methylene chloride, 2.42 g (28.5 mmol) of 1-methylpyrrolidine was added in one portion at room temperature. 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.

IC: C cm ^-1

400, 1780 (s), 1740, 1675, 1530.

Thin layer chromatography:

ethanol-CHCl3 (1: 3),

Rf = 0.30 (Rf = 0.95 for VIIa ‘).

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 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 into 400 mL of ethyl acetate and filtered to give 8.08 g of a solid which was a mixture of the title compound and the corresponding 4 ² -isomer (Λ ³ / Δ ² = 1: 8) (by high pressure liquid chromatography, Lichrosorb). RP-18 10-15% methanol in 0.01M phosphate buffer, pH 7). A second batch of 28.9 g (31.0 mmol) of VIIa 'yielded 16.0 g of crude product (Δ ³ / Δ ² = 1: 8). Isolation of the desired ^β- isomer from the combined crude products (24.08 g) using preparative high pressure liquid chromatography (System 500, Waters Associates, Prep PAK 500 / C18, 5-10% CH 3 OH) afforded 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)

7 - [(Z) -2-methoxyimino-2- (2-amino-7 - ((Z) -2-methoxyimino-2- [2- (2-aminothiazol-4-yl) acetahido]) - 3 - [( 1-Methyl 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 the ratio Δ ³ / Δ ² in the reaction product.

A solution of N-methylpyrrolidine (0.01 mL, 0.087 mmol) in ether (0.1 mL) was added to a suspension of 3-iodomethyldervate of formula VIIa '(45 mg, 0.048 mmol) in the indicated amount in the solvent and stirred. The reaction mixture was diluted with ether (5 mL) and the resulting precipitate was filtered off and treated with 90% trifluoroacetic acid, stirred for one hour and then evaporated to dryness under reduced pressure at a temperature below 20 ° C. Δ ³ / Δ ² in the product obtained is determined by high pressure liquid chromatography (Lichrosorb RP-18, mobile phase, 0,01 M ammonium phosphate buffer pH 7,2 containing 15% НзОН, retention time Δ ³ = 6,60 minutes Δ ² 5,56 The yield of product and the ratio of Δ ³ / Δ ² isomers for each experiment is given below.

, 26

try T <^ 2 ^] ^ oius1t & dlo · Vila ratio "(g) reaction yield ratio C. to solvent (ml) time (%) 3 / Δ ² • (min) 1 CH2Cl2 1: 20 15 Dec 73 1/8 2 C ^ Clž · - ^ (1/10) 1: 100 15 Dec 25 4/1 3 ethyl acetate-ether (1/10) 1: 100 15 Dec '' 27 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

Example 4

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

A. B-enzhydryl 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 (IV b) (1.095 g, 2.4 mol) in dichloromethane (20 mL) was added phosphorus pentachloride (500 mg). . After stirring at room temperature for 1 hour, the mixture was added all at once to an ice-cooled solution of compound V (1.083 g,

2.4 mmol) and bis (trimethylsilyl) acetamide (1 ml) in dichloromethane (20 ml) After stirring for half an hour, the reaction mixture was poured into 10% aqueous sodium bicarbonate (200 ml) and extracted with chloroform (100 ml). The extract was washed with water, dried (MgSO4) 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: δ ^CO ™ ppm

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

3.53 (2H, ABq, 2-CH 2),

4.37 (2H, a, - CH 2 Cl 2),

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

4.90 (1 H, d, 6-H),

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

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

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

B. Diphenylmethyl 7 - [(Z) -2-ethoxyimino-2-

(2-tritylaminothiazol-4-yl) acetamido] -3-iodomethyl-3-cephem-4-carboxylate (VIIIb)

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. The mixture was filtered and the filtrate was poured into water and extracted with ethyl acetate. The organic phase was washed successively with 5% aqueous Na2SO4 solution, water and saturated aqueous sodium chloride solution, dried over magnesium sulfate and evaporated to give 1.04 g (89%) of VIIb.

NMR: δ cdg. pp _m

3.55 (2H, q, 2-CH 2),

4.27 (2H, s, □ №--),

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

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

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

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

C. 7-f (Z) -2-Ethoxyimido-242-aminothiazol-4-yl) acetamido] -3 - [(1-methyl-4-l-pyrrolidinium) phenyl] - -3-tert-fem-4-carboxylate (1b)

A mixture of VIIb (333 mg, 0.35 mmol) and N-methylpyrrolidine (00 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% aqueous 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 off 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 lyophilization yielded 0.062 g of composition A ² and Δ ³ isomer (Δ2: Δ3 = 5: 1). The mixture was purified by high pressure liquid chromatography (Lichrosorb RP-18, 8 x 300 mm, 15% methanol) and the desired is3 isomer (1b) was isolated as a pale yellow powder, 4.9 mg [2.7%].

UV _: λ phosphate buffer, pH 7

235 [15 ^1000]

258 (14,000).

NMR:? D20 ppm

1.43 (3H, t).

2.33 (4 H, 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-propoxylmino) -2- (2-aminothiazol-1-yl) ac etamido] -3 - [[1-methyl-1-methyl-pyridinium) methyl] -3 -cephene-4-carboxylate (Ic) 1 * S 1 2

A. Diphenylmethyl 3-chloromethyl-7 - [(Z) -2- (2-propoxyimino) -2- (2-tritylaminophenol-4-yl) ace tamido] -3-cephem-4-carboxylate )

Mixture of (Z) -2- (2-propoxyimino) -2- (2-trityl-4-yl) -Ο- ^, Ζ1-4Ζ-4-γ1) ': ον €! of acid (IVc) (707m), 1.5mmol) and phosphorus pentachloride (344mg, 1.65mmol) in dichloromethane (14mL) was stirred at room temperature for one hour and then poured into a solution of compound V (677mg) , 1.5 mmol) and bis (trimethylsilyl) acetamide (1.1 mL, 4.5 mmol) in dichloromethane (15 mL). 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 compound VI.

ICI: cm ^_1

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, 1,090, 1,060, 1,010, 990,

840, 710, 700.

UV: 1 µm (.)

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

NMR: O · ¹ · ¹ [ppm ·

1.35 (6H, d, | -6Hz)

3.50 (2H, s);

4.35 (2 H, s),

4.58 (lil, m, J = 6Hz),

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

5.91 (1H, d-d, J = 4.5, 9Hz, d, JO),

4.5Hz),

6.68 (1 H, s),

6.88 (1 H, s),

7.25 (25H, col.

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

A mixture of Compound VIc (500 mg, 0.55 mmol) and sodium iodide (248 nm, 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 successively with 10% aqueous sodium thiosulfate solution (10 ml), water (10 ml) and brine (10 ml), dried over sodium sulfate and evaporated to give 494 mg ( 90%) of the title compound (Fairy).

IC: cm ^-1

360, 3,040, 3,020, 2,960, 1,785, 1,720,

680, 1,600, 1,520, 1,500, 1,450, 1,370.

300, 1,230, 1,150, 1,115, 1,080, 990,

900, 840, 750, 700.

UV: λ ^Et ° nm. (ε)

240 (24,900), 260 (19,400).

NMR: δ ^{CDCl 3} ppm

1.30 (6'H, d, J 6Hz),

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

4.22 (2 H, s),

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

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

5.83 (1H, d-d, J 4.5 and 9Hz, d 6 O),

6.66 (1H, 3),

6.87 (1 H, s),

7.25 (25 H, s).

C. 7- [(Z) -2- (2lproρoxyimino) -2- (2-amtoo · thiaznl-4-yl) acetate <AMTD [| 3- ^{((1-methyllllpyrrofid.i,} niπm) methyl] -3 - [alpha] 14-carboxylate (1c)

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 is filtered off. A solution of the precipitate in 90% trifluoroacetic acid (4.5 ml) was stirred for 30 minutes at room temperature and then evaporated in vacuo. This was triturated with ether to give 317 mg of a 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 by 1 press water to give 109 mg of a mixture of Δ ² and Δ ⁵ isomers (A ² / A3 6: 1). 100 mg of this mixture with Purify by high pressure liquid chromatography (Lichrosorb RP-18, 15% McOH) to give 5 mg (3%) of the desired compound 1c.

UV: J * J ^lurr nm (e)

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

NMR: .delta

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 (1H, d, J 4.5 Hz),

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

7.05 (II, col.

Example 6

7- ((Z) -4-allyloxylmino-z- (2-amino-thiazol-4-yl) acetamido) -3 - ((1-methyl-1-pyrooidinidinium (methyl) -3-methyl) -4-carboxylate (1d)

A. Benzhydryl 7- ((Z) -2-allyloxy-amino) -2- (2-trityl-amino-4- (4-arylamino) -4-yl) acetamido] -3-chloro ^, thyl-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 24-512 ° C for 30 min until clear. A solution of (ZJ-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 ( The mixture was stirred for 20 minutes at room temperature and diluted with ethyl acetate (200 ml), washed with aqueous sodium bicarbonate solution and water, dried and evaporated. 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 and evaporated under reduced pressure to give the title compound (Vid). in the form of an amorphous powder; obstructions, 2.32 g (89%). mp. 100-115 ° C (with decomposition.

IC: cm ^-1

990, 1790, 1730, 1680, 1530, 1380, 1250, 1160, 1020.

NMR: <5 ^{cm -1 13} ppm

3.50 (2 H, 2-H),

4.32 (2H, s, 3-CH 2),

4,6 - 6,1 (7H, m, НгСН = СН2 and 6,7-H),

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

6.90 (1 H, s, PhsCH),

7.1 - 7.6 (ZO, m, phenyl protons).

Analysis for C18H40N5O5S2Cl. 1/3 СНС1з: calculated:

C 64.05, H 4.45, N 7.73, S 7.08, Cl 7.82, found ·

C 64.13, 63.99, H 4.61, 4.64, N 7.50, 7.30, S 6.85, 6.85, Cl 7.55, 7.46.

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, 13.3 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 the compound of formula VIId 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), and precipitated. This gave the quaternized product (yield 500 mg). A mixture of the quaternized product and trifluoroacetic acid (2 mL) was allowed to stand at room temperature

1.5 hours, diluted with ether, precipitated trc crude product trifluoroacetic acid 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 amorphous powder (yield 124 mg) containing the desired product (17%) and the corresponding Δ ² isomer (83 why.). The mixture was purified by high pressure liquid chromatography [Lichrosorb RP-18; 0.01 M NH 4 H 2 PO 3 (pH 7): Nitro-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.).

IC: cm * ¹

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

200

UV: v ^ _x ⁷ i ^Klfr nm (ε)

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

NMR: δ ^{D 20} ppm

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

3.10 (3H, s, NCH 3),

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

5.95 (1 H, d, 4Hz, -H),

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

P 14 to 1 and d 7

7- (2- (2-aminothiazo-4-yl) - (Z) -2- (2-carboxyprop-2-oxyimidino) acetamido) -3 - [(1-methyl-1-pyrrolidinium) methyl] 1 -3-Cephem-4-carboxylate (1e)

A. Benzhydryl 3-chloromethyl-7- [(Z) -2- ^(2-! T butoxykarbonylprop-2-oxyimino] -2- (2-trityl-amino-4 thiaZ'Ol 1-yl) acetamido-3- one cefem-4-carboxylate (Va)

Procedure 1

A mixture of [Z] -2- (tert -butoxycarbinylprop-4-yloxyimino) -2- (2-tritylammothiazol-4-yl) acetic acid (Pα) (1.94 g, 3.6 mmol), DDC [ 742 mg, 3.6 mmol) and N-hydroxybenztr bean (486 mg, 3.6 mmol) in tetrahydrofuran (45 mL) were stirred at room temperature for 45 minutes while dicyclohexylurea was precipitated. The dicyclohexylurea was filtered off and the filtrate was treated with compound V (1.5 g, 3.6 mmol). 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 chloride solution, dried over magnesium sulfate and evaporated to dryness. The residue (3.9 g) was dissolved in n-hexane / chloroform (1: 2) and passed through a column of silica gel (40 g) in the same solvent mixture. The fractions containing the title compound were evaporated in vacuo to give 1.3 g (39%) of compound Va, m.p. &gt; 100 ° C (dec.).

IR: cm -1

990,1791; 1715, 1690.

UV: λ 240 (E1 · 280), 265 (Ej 190).

NMR: Op

1.45 (9H, s);

1.63 and 1.66 (ro 6H, «),

3.49 (2H, broad s),

4.34 (2 H, s),

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

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

6.66 (1 H, sl,

6.86 (1H, s),

7.0 - 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 (trimethylsilyliacetamide (3.33 mL, 13.5 mmol) at room temperature for 50 min to give To this solution was added an acid chloride prepared from compound 11a '(2.56 g, 4.49 mmol) and phosphorus pentachloride (1.12 g, 5.38 mmol) in 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). It is dried and evaporated from Na 2 O 240 (E - 270), 265 ml, and the evaporated syrup (5 g) is chromatographed on a silica gel column (100 g) eluting with 10: 1-toluene / ethyl acetate. Fractions containing the desired compound were combined and evaporated to give 2.84 g (35%) of Compound Va.

B. Benzhydryl 7- [(Z) -2- (2-tert-butoxycarbonynyl prop-1-oxyimino) -2- (2-tritylaminothiazol-4-yl) acetamido · | 3-Iodomethyl-3-cephem-4-carboxylate (VIa)

A mixture of Compound Va (500 mg, 0.53 mmol) and Nal (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 was added methylene chloride (20 mL) and water (10 mL). The organic phase is washed with 10% sodium thiosulphate (5 ml) and brine (5 ml), dried over magnesium sulphate and evaporated in vacuo to give 540 mg (90%) of compound VIa as an amorphous powder, m.p. 106 ° C (dec.).

IR: too cm -1

35oj 1,790, 1,690.

(E 1-190).

NMR: Sppm

1.44 (9H, s);

1.65 (6H, s);

3.54 (1 H) 1

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 - 7.5 (25H, m).

AND

C. 7- (2- (2-Aminothiazol-4-yl) - (Z) -2- (2-carboxyprop-2-oxyimino) acetumido] -3 - [(1-methyl-1-pyrrobdinium) methyl] -3-Cephem-4-carboxylate (to)

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 is then evaporated to dryness. The crude salt of compound Ia with trifluoroacetic acid (245 mg, quantitative yield) is obtained, which is a 1: 4 mixture of the A ³ - and A 2 isomers. 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 addition of 1M 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. Mp 160 ° C (dec.).

Company ID:

400, 1775, 1610.

_UV: λ ^fo SFAT ° Pf characterized ⁱⁿ pH 7 nm _(ε

237 (15,700),

257 (155,500).

NMR: δ ^-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-oxylmino) -2- (2-tritylaminothiazol-4-yl) acetic acid was replaced with an equimolar amount of (Z). -2- (1-tert-butoxycarbonylmethoxy-amino) -2- (2-tritylamino-thiazol-4-yl) -acetic acid; 4-yl) acetic acid, (Z) -2- (2-tert-butoxycarbonylbut-2-oxyimino) -2- (2-tritylaminothiazol-4-yl) acetic acid, (Z) -2- (3- tert-butoxycarbonylpiperazin-3-oxoimino) -1- (2-tritylaminothiazol-4-yl) acetic acid, (Z) -2- (1-tert-butoxycarbonylcyclopropyl-1-oxyimino) - 2- (2-tritylaminothiazol-4-yl) -acetic acid, (Z) -2- (1-tert-butoxycarbonylcyclobut-1-oxyimino) -2- (2-trityl-aminothlazoM2) 4 3 5 1 2

and (Z) -2- (1-tert-butoxycarbonylcyclopent-1-oxyimino) -2- (2-tritylamino) hiazol-1-yl (acetic acid) to give

7- (2- (2-aminothiazol-4-yl) - (Z) -2- (carboxy • ^met'hoxyimi: No ·) acetamldoi] -3- [(1-methyl-pyrrolidone olidinium jmethyl] - 3-cep-1- (2-carboxylate),

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

7- [2- (2-aminothiazol-4-yl) - (Z) -2- (2-k-arboxybut-2-oxyimino) acetamido] -3 - [(1-methyl-1-ipyrrolidimum) methyl ] -3-cephem-4-carboxylate,

7- (2- ^{(2-aminotbiazo;} L-4-yl) - (Z) -2- (3-carboxypent-3-oyyimmo jacetamido] -3- [(l-methyl-l-pyrrolidinium) metbyl] -3 -cephem-4зcarboxy compounds,

7- (2- (2-aminothiazol-4-yl) - (Z) -2- (1-carboxycyclo) irop-1-oxyimino Jacetamido) -3 - [(1-methylilpyrrolidinium) methyl] - 3-cep-4-carboxylate,

7- [2- (2-aminO'tbiazoi-4-yl) - (Z) -2- (1-carboxycyclobut-l-oxyimino) acetamido] -3- [(lm-l ^ι etbyl-pyrrolidlnium) methyl] - 3-cephem-4-carboxylate, a

7- (2- (2-aminotbiazol-4-yl) - (Z) -2- (1-karboyycykloρeint ^l-oxyl-ι amino] acetamido] -3- [(l ^^^ ^^^ if ^ l - (1-methyl) -3-cefeim-4-carboxylate.

Claims (3)

A process for the preparation of cephalosporin derivatives of the general formula I (I) having 1 to 4 carbon atoms, allyl, 2-butenyl or 3-butenyl or a group wherein: R ² is straight or branched alkyl л с κ j 2 R3-C-R4 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 may form a cycloalkylidene ring containing from 3 to 5 carbon atoms, or non-toxic pharmaceutically acceptable salts physiologically hydrolyzable esters or solvates thereof, characterized in that acylating a compound of formula XVI (XV1) or an N-derivative thereof, wherein B ¹ is a hydrogen atom or a common carboxyl protecting group acylating acid derivative of formula XVII C COOH II OR 2 where R ² is a conventional amino protecting group, R ² is as defined above, and the optionally present carboxyl group in the substituent R ² is protected to form a compound of formula XV wherein Β ¹ , B ² and R ² are as defined above The compounds of formula (I) are optionally converted into non-toxic pharmaceutically acceptable salts, physiologically hydrolyzable esters or solvates. 2. A method according to claim 1, characterized in that the reacting together corresponding compounds of general formulas • XVI and XVII to give 7- [2- (2-aminothiazol-4-yl] - (Z) -2- (2 ^l karboxyρro O-S-oxoimino) -acetamido] -3 - [(1-methyl-1-pyrrolidinium) methyl] -3-cephem-4-carboxylate or a non-toxic pharmaceutically acceptable salt, physiologically hydrolyzable ester or solvate thereof. 3. A process according to claim 1, wherein the corresponding compounds of formulas XVI and XVII are reacted to form a compound of formula I, wherein: R 2 is methyl, ethyl, isopropyl or allyl or a non-toxic pharmaceutically acceptable salt or solvate thereof.