SIDEROPHORE CEPHALOSPORIN CONJUGATES AND USES THEREOF

Abstract

A compound of compound according to formula (I):

##STR00001##

wherein X is selected from the group consisting of CH, CCl and N, Z is selected from the group consisting of CH.sub.2COOH, CH(CH.sub.3)COOH, C(CH.sub.3).sub.2COOH and CH.sub.2F, D is a single bond connecting A and Ar or selected from the group consisting of CO, NHCO and N(C.sub.0-6)alkyl-CO, A is selected from the group consisting of a (C.sub.1-6)alkanediyl and a (C.sub.3-6)cycloalkanediyl or, if D is N(C.sub.0)alkyl-CO, A forms a 4- to 7-membered aliphatic heterocyclic ring with the nitrogen atom of N(C.sub.0)alkyl-CO in D, and Ar is a 6-membered aromatic ring with a first hydroxyl group in para-position to D, a second hydroxyl group in meta-position to D, and with at least one electron-withdrawing element
and uses thereof.

Claims

1. A compound according to formula (I): ##STR00023## wherein X is selected from the group consisting of CH, CCl and N, Z is selected from the group consisting of CH.sub.2COOH, CH(CH.sub.3)COOH, C(CH.sub.3).sub.2COOH and CH.sub.2F, D is a single bond connecting A and Ar or selected from the group consisting of CO, NHCO and N(C.sub.0-6)alkyl-CO, A is selected from the group consisting of a (C.sub.1-6)alkanediyl and a (C.sub.3-6)cycloalkanediyl or, if D is N(C.sub.0)alkyl-CO, A forms a 4- to 7-membered aliphatic heterocyclic ring with the nitrogen atom of N(C.sub.0)alkyl-CO in D, and Ar is a 6-membered aromatic ring with a first hydroxyl group in para-position to D, a second hydroxyl group in meta-position to D, and with at least one electron-withdrawing element.

2. The compound according to claim 1, wherein the electron-withdrawing element may be a heteroatom in the aromatic ring or an electron-withdrawing substituent at the aromatic ring.

3. The compound according to claim 1, wherein the compound is a compound according to formula (II) ##STR00024## wherein X, Z, D, and A are defined as in claim 1 and wherein E is selected from the group consisting of CH, N, N.sup.+—CH.sub.3, and N.sup.+—O.sup.−, and G is H or an electron-withdrawing substituent, with the provision that if E is CH, D is selected from the group consisting of CO, NHCO and N(C.sub.0-6)alkyl-CO and G is an electron-withdrawing substituent.

4. The compound according to claim 3, wherein G is an electron-withdrawing substituent selected from the group consisting of F, Cl and OCF.sub.3.

5. The compound according to claim 1, to wherein a shortest linking path in A is defined by the number of bonds extending between the O of the oxime group and D excluding the bond to the O of the oxime group and excluding the bond to D or the single bond of D, and A is selected such that the shortest linking path in A is shorter than or equal to 3 covalent (C—C) single bonds, in particular shorter than 3 acyclic covalent (C—C) single bond, and preferably shorter than or equal to 2 covalent (C—C) single bonds, in particular shorter than 2 acyclic covalent (C—C) single bonds.

6. The compound according to claim 5, wherein the shortest linking path in A extends over no single (C—C) bond, one cyclic or acyclic (C—C) single bond, or two cyclic covalent (C—C) bonds.

7. The compound according to claim 1, wherein A is selected from the group consisting of methanediyl, ethane-1,2-diyl, propane-1,3-diyl, propane-1,2-diyl, cyclobutane-1,3-diyl, cyclopentane-1,3-diyl, cyclohexane-1,3-diyl, azetidine-1,3-diyl, pyrrolidine-1,3-diyl and piperidine-1,3-diyl, wherein in azetidine-1,3-diyl, pyrrolidine-1,3-diyl and piperidine-1,3-diyl, D is N(C.sub.0)alkyl-CO and the nitrogen atom of N(C.sub.0)alkyl-CO in D is the nitrogen in the heterocyclic ring, preferably selected from the group consisting of methanediyl, ethane-1,2-diyl, cyclobutane-1,3-diyl, and pyrrolidine-1,3-diyl with D being N(C.sub.0)alkyl-CO and the nitrogen atom of N(C.sub.0)alkyl-CO in D being the nitrogen in the pyrrolidine ring.

8. The compound according to claim 3, wherein D is NHCO or N(C.sub.0-6)alkyl-CO, A is a (C.sub.1-6)alkanediyl, preferably a (C.sub.1-4)alkanediyl, such as in particular the (C.sub.2)alkanediyl ethane-1,2-diyl, a (C.sub.3-6)cycloalkanediyl, preferably (C.sub.4-5)cycloalkanediyl, such as in particular the (C.sub.4)cycloalkanediyl cyclobutane-1,3-diyl, or A forms a 4- to 7-membered aliphatic heterocyclic ring with the nitrogen atom of N(C.sub.0)alkyl-CO in D, preferably a 5-membered aliphatic heterocyclic ring, such as in particular the 5-membered heterocyclic ring pyrrolidine-1,3-diyl with the nitrogen atom of N(C.sub.0)alkyl-CO in D being the nitrogen in the pyrrolidine, E is CH, and G is Cl.

9. The compound according to claim 3, wherein D is a single bond, A is a (C.sub.1-6)alkanediyl, preferably a (C.sub.1-4)alkanediyl, such as methanediyl, or a (C.sub.3-6)cycloalkanediyl, E is N.sup.+—O.sup.−, and G is H.

10. The compound according to claim 1, wherein -A-D-Ar is selected from the group consisting of ##STR00025## preferably from the group consisting of ##STR00026## more preferably is selected from the group consisting of ##STR00027##

11. The compound according to claim 1, wherein X is CH or N.

12. The compound according to claim 1, wherein Z is C(CH.sub.3).sub.2COOH or CH.sub.2F.

13. The compound according to claim 1 selected from the group consisting of (6R,7R)-7-[[(2Z)-2-(2-aminothiazol-4-yl)-2-(1-carboxy-1-methyl-ethoxy)imino-acetyl]amino]-3-[(1,5-dihydroxy-4-oxo-2-pyridyl)methoxyiminomethyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, (6R,7R)-7-[[(2Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methyl-ethoxy)imino-acetyl]amino]-3-[(1,5-dihydroxy-4-oxo-2-pyridyl)methoxyiminomethyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, (6R,7R)-7-[[(2Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)-2-(fluoromethoxyimino)acetyl]amino]-3-[(1,5-dihydroxy-4-oxo-2-pyridyl)methoxyiminomethyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 7-[[(2Z)-2-(2-aminothiazol-4-yl)-2-(1-carboxy-1-methyl-ethoxy)imino-acetyl]amino]-3-[2-[(2-chloro-3,4-dihydroxy-benzoyl)amino]ethoxyiminomethyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 7-[[(2Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methyl-ethoxy)imino-acetyl]amino]-3-[2-[(2-chloro-3,4-dihydroxy-benzoyl)amino]ethoxyiminomethyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 7-[[(2Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)-2-(fluoromethoxyimino)-acetyl]amino]-3-[2-[(2-chloro-3,4-dihydroxy-benzoyl)amino]ethoxyiminomethyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, 7-[[(2Z)-2-(2-aminothiazol-4-yl)-2-(1-carboxy-1-methyl-ethoxy)imino-acetyl]amino]-3-[[3-[(2-chloro-3,4-dihydroxy-benzoyl)amino]-trans-cyclobutoxy]iminomethyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid and 7-[[(2Z)-2-(2-aminothiazol-4-yl)-2-(1-carboxy-1-methyl-ethoxy)imino-acetyl]amino]-3-{[(3S)-1-(2-chloro-3,4-dihydroxy-benzoyl)pyrrolidine-3-yl]oxyiminomethyl}-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid.

14. A composition comprising a compound of claim 1 in association with at least one pharmaceutical excipient, optionally comprising further at least one pharmaceutical active agent.

15. The compound according to claim 1, wherein the compound is used as medicine.

16. The compound according to claim 1, wherein the compound is used for treatment and prevention of an infectious disease mediated by microbes or bacteria, preferably Gram-negative bacteria.

17. The compound according to claim 16, wherein the infectious disease is selected from the group consisting of a respiratory tract infection of the lower and/or upper respiratory tract, an urinary tract infection, an intraabdominal infection, a systemic infection, a prosthetic joint infection, a gastrointestinal infection, and an infection of skin and/or soft tissue, wherein an upper and lower respiratory tract infection includes cystic fibrosis and bronchiectasis.

18. The compound according to claim 16, wherein the infectious disease is mediated by a bacteria selected from the group consisting of Escherichia coli, Salmonella typhimurium, Citrobacter freundii, Klebsiella pneumoniae, Enterobacter cloacae, Enterobacter aerogenes, Moraxella catarrhalis, Proteus mirabilis, Proteus vulgaris, Providencia stuartii, Serratia marcescens, Acinetobacter baumannii, Acinetobacter lwoffi, Pseudomonas aeruginosa, Haemophilus influenzae, Burkholderia cepacia, Burkolderia cenocepatia, Burkholderia vietnamensis, Streptococcus spp., and Streptococcus pneumoniae, preferably selected from the group consisting of Escherichia coli, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Burkholderia cepacia.

19. A method of treating an infectious disease in a subject in need thereof, comprising administering an effective amount of a compound according to formula (I) ##STR00028## to the subject, wherein X is selected from the group consisting of CH, CCl and N, Z is selected from the group consisting of CH.sub.2COOH, CH(CH.sub.3)COOH, C(CH.sub.3).sub.2COOH and CH.sub.2F, D is a single bond connecting A and Ar or selected from the group consisting of CO, NHCO and N(C.sub.0-6)alkyl-CO, A is selected from the group consisting of a (C.sub.1-6)alkanediyl and a (C.sub.3-6)cycloalkanediyl or, if D is N(C.sub.0)alkyl-CO, A forms a 4- to 7-membered aliphatic heterocyclic ring with the nitrogen atom of N(C.sub.0)alkyl-CO in D, and Ar is a 6-membered aromatic ring with a first hydroxyl group in para-position to D, a second hydroxyl group in meta-position to D, and with at least one electron-withdrawing element.

20. The method according to claim 19, wherein the infectious disease is mediated by microbes or bacteria, preferably Gram-negative bacteria.

21. The method according to claim 19, wherein the infectious disease is selected from the group consisting of a respiratory tract infection of the lower and/or upper respiratory tract, an urinary tract infection, an intraabdominal infection, a systemic infection, a prosthetic joint infection, a gastrointestinal infection, and an infection of skin and/or soft tissue, wherein an upper and lower respiratory tract infection includes cystic fibrosis and bronchiectasis.

22. The method according to claim 19, wherein the infectious disease is mediated by a bacteria selected from the group consisting of Escherichia coli, Salmonella typhimurium, Citrobacter freundii, Klebsiella pneumoniae, Enterobacter cloacae, Enterobacter aerogenes, Moraxella catarrhalis, Proteus mirabilis, Proteus vulgaris, Providencia stuartii, Serratia marcescens, Acinetobacter baumannii, Acinetobacter lwoffi, Pseudomonas aeruginosa, Haemophilus influenzae, Burkholderia cepacia, Burkolderia cenocepatia, Burkholderia vietnamensis, Streptococcus spp., and Streptococcus pneumoniae, preferably selected from the group consisting of Escherichia coli, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Burkholderia cepacia.

23. The method according to claim 19, wherein the effective amount of the compound according to formula (I) is administered in the form of a pharmaceutical composition.

Description

EXAMPLES

[0108] Herein, including the examples and the reaction scheme the following abbreviations are used:

TABLE-US-00002 .sup.1H-NMR proton nuclear magnetic resonance spectroscopy ° C. degrees Celsius BOC tert-Butyloxycarbonyl BSA N,O-Bis(trimethylsilyl)-acetamide DMSO dimethylsulfoxide EDC N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride eq equivalents h hour(s) HOBT 1-hydroxybenzotriazole HPLC high performance liquid chromatography MS mass spectrometry N normality NMR nuclear magnetic resonance spectroscopy m/e mass/charge ratio THF tetrahydrofuran

Example 1

(6R,7R)-7-[[(2Z)-2-(2-aminothiazol-4-yl)-2-(1-carboxy-1-methyl-ethoxy)imino-acetyl]amino]-3-[(1,5-dihydroxy-4-oxo-2-pyridyl)methoxyiminomethyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid

Scheme 2

[0109] ##STR00011##

Step 1: Amide Formation

[0110] To an ice cooled solution of compound B (WO95/29182) (1.47 g, 6.44 mmol) and BSA (3.15 mL, 2 eq) in acetonitrile (10 mL) was added a suspension of compound A1 (U.S. Pat. No. 4,500,526) (3.8 g, 1 eq) in acetonitrile (20 mL). The resulting reaction mixture was stirred for 1 h under ice cooling, for 1 h at room temperature and then added slowly to ice water. The resulting precipitate was filtered and dried under reduced pressure to obtain compound C1 as yellow solid which was directly used for the next step.

[0111] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, δ, ppm): 9.46 (d, 1H, CONH, J=8.2 Hz), 7.43-7.18 (m, 15H, trityl-H), 6.71 (s, 1H, thiazole-H), 6.30, 6.22 (2xs, 1H, O—CH—O), 5.90 (dd, 1H, H-6, J=5.2 Hz, 8.2 Hz), 5.16 (d, 1H, H-7, J=5.2 Hz), 3.76 (d, 1H, H-2a, J=18.6 Hz), 3.58 (d, 1H, H-2b, J=18.6 Hz), 1.41 (s, 6H, 2xCH.sub.3), 1.37 (s, 9H, COO.sup.tBu).

[0112] MS m/e: 782 [M+H].sup.+

Step 2: Oxime Formation

[0113] To a suspension of compound C1 (Example 1 step 1) and compound D1 (EP0430286) (1.11 g, 1 eq) in acetonitrile (15 mL) was added under ice cooling 1 N hydrochloric acid (19.3 mL, 3 eq). The resulting mixture was allowed to warm to room temperature, stirred for 3 h and then slowly added to ice water. The resulting precipitate was filtered off and dried under reduced pressure to obtain compound I1 in protected form as a solid which was directly used for the next step.

[0114] .sup.1H-NMR (400 MHz, DMSO-d.sub.6, δ, ppm): 9.41 (d, 1H, CONH, J=8.1 Hz),), 8.36 (s, 1H, azomethine), 7.80 (s, 1H, dihydropyridine H-6), 7.38-7.17 (m, 15H, trityl-H), 6.74 (s, 1H, dihydropyridine H-3), 6.68 (s, 1H, thiazole-H), 5.79 (dd, 1H, H-6, J=5.0 Hz, 8.1 Hz), 5.23 (d, 1H, H-7, J=5.0 Hz), 5.15 (s, 2H, O—CH.sub.2), 3.85 (d, 1H, H-2a, J=18.0 Hz), 3.58 (d, 1H, H-2b, J=18.0 Hz), 1.38 (bs, 6H. 2xCH.sub.3), 1.35 (s, 9H, COO.sup.tBu).

[0115] MS m/e: 936 [M+H].sup.+

Step 3: Deprotection

[0116] Compound I1 in protected form (example 1 step 2) was dissolved in trifluoracetic acid (20 mL) under ice cooling. The resulting mixture was stirred under ice cooling for 4 h and then slowly added to cold diethyl ether. The resulting precipitate was filtered off and dried under reduced pressure to obtain compound I1 as a solid in the form of a trifluoroacetate salt.

[0117] .sup.1H-NMR (400 MHz, MeOH-d.sub.4, δ, ppm): 8.46 (s, 1H, azomethine), 7.84 (s, 1H, dihydropyridine H-6), 6.94 (s, 1H, thiazole-H), 6.84 (s, 1H, dihydropyridine H-3), 5.87 (d, 1H, H-6, J=5.1 Hz), 5.20 (s, 2H, O—CH.sub.2), 5.16 (d, 1H, H-7, J=5.1 Hz), 3.89 (d, 1H, H-2a, J=18.2 Hz), 3.50 (d, 1H, H-2b, J=18.2 Hz), 1.52 (s, 3H, CH.sub.3), 1.51 (s, 3H, CH.sub.3).

[0118] MS m/e: 638 [M+H].sup.+

Step 4a (Optional): Salt Exchange and Chromatography

[0119] Compound I1 (1.6g, 1.85 mmol) in the form of a trifluoroacetate salt (Example 1 Step 3) was suspended in water and a 1N NaOH was slowly added until a pH of 6 to 6.5 was reached. To the resulting solution 1N HCl was added until a pH of 2 was reached and the resulting precipitate was filtered off and subjected to reversed phase chromatography obtain compound I1 after freeze-drying, as an off-white solid in the form of a hydrochloride salt (530 mg).

[0120] .sup.1H-NMR (400 MHz, MeOH-d.sub.4, δ, ppm): 8.45 (s, 1H, azomethine), 7.77 (s, 1H, dihydropyridine H-6), 6.81 (s, 1H, thiazole-H), 6.77 (s, 1H, dihydropyridine H-3), 5.87 (d, 1H, H-6, J=5.0), 5.18 (s, 2H, O—CH.sub.2), 5.14 (d, 1H, H-7, J=5.0 Hz), 3.86 (d, 1H, H-2a, J=18.0 Hz), 3.48 (d, 1H, H-2b, J=18.0 Hz), 1.52 (s, 3H, CH.sub.3), 1.50 (s, 3H, CH.sub.3).

[0121] MS m/e: 638 [M +H].sup.+

[0122] The compounds of formula (III) wherein R, X and Z are as defined in Table 2 (Examples 2 to 8) are prepared according, e.g. analogously to a method conventional or as set out under Example 1 Step 2 to 3, and optionally Step 4 above, but using appropriate starting materials.

##STR00012##

TABLE-US-00003 TABLE 2 Ex- am- ple Name/Characterization R X Z 2 (6R,7R)-7-[[(2Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)-2-(1- N C(CH.sub.3).sub.2COOH carboxy-1-methylethoxy)imino-acetyl]amino]-3-[(1,5-dihydroxy- 4-oxo-2-pyridyl)methoxyiminomethyl]-8-oxo-5-thia-1-azabicyclo [4.2.0]oct-2-ene-2-carboxylic acid in the form of a hydrochloride .sup.1H-NMR (400 MHz, MeOH-d.sub.4, δ, ppm): 8.57 (s, 1H, azomethine), 7.92 (s, 1H, dihydropyridine H-6), 6.93 (s, 1H, dihydropyridine H-3), 6.01 (d, 1H, H-6, J = 5.1 Hz), 5.31 (s, 2H, O—CH.sub.2), 5.26 (d, 1H, H-7, J = 5.1 Hz), 4.00 (d, 1H, H-2a, J = 18.2 Hz), 3.60 (d, 1H, H-2b, J = 18.1 Hz), 1.64 (s, 3H, CH.sub.3), 1.63 (s, 3H, CH.sub.3). MS m/e: 639 [M + H].sup.+ 3 (6R,7R)-7-[[(2Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)-2-(fluoro- methoxyimino)acetyl]amino]-3-[(1,5-dihydroxy-4-oxo-2-pyridyl) methoxyiminomethyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxylic acid in the form of a trifluoroacetate .sup.1H-NMR (400 MHz, MeOH-d.sub.4, δ, ppm): 8.56 (s, 1H, azomethine), 8.28 (s, 1H, dihydropyridine H-6), 7.22 (s, 1H, dihydropyridine H-3), 6.00 (d, 1H, H-6, J = 5,1 Hz), 5.82 (d, 2H, O—CH.sub.2—F, J = 54.6 Hz), [00013] N CH.sub.2F 5.42 (s, 2H, O—CH.sub.2), 5.28 (d, 1H, H-7, J = 5.1 Hz), 4.00 (d, 1H, H-2a, J = 18.2 Hz), 3.62 (d, 1H, H-2b, J = 18.2 Hz). MS m/e: 583 [M − H].sup.− 4 7-[[(2Z)-2-(2-aminothiazol-4-yl)-2-(1-carboxy-1-methylethoxy) imino-acetyl]amino]-3-[2-[(2-chloro-3,4-dihydroxybenzoyl) amino]ethoxyimino methyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct- 2-ene-2-carboxylic acid in the form of a hydrochloride .sup.1H-NMR (400 MHz, MeOH-d.sub.4, δ, ppm): 8.42 (s, 1H, azomethine), 6.92 (s, 1H, thiazole-H), 6.84 (d, 1H, arom. H-6, J = 8.2 Hz), 6.74 (d, 1H, arom. H-5, J = 8.2 Hz), 5.95 (d, 1H, H-6, J = 5.0 Hz), 5.24 (d, 1H, H-7, J = 5.0 Hz), 4.27 (t, 2H, O—CH.sub.2, J = 5.4 Hz), 4.03 (d, 1H, H-2a, J = 18.1 Hz), 3.67-3.55 (m, 3H, H-2b, N—CH.sub.2), [00014] CH C(CH.sub.3).sub.2COOH 1.62 (s, 3H, CH.sub.3), 1.60 (s, 3H,CH.sub.3). MS m/e: 710 [M − H].sup.− 5 7-[[(2Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methyl- ethoxy)imino-acetyl]amino]-3-[2-[(2-chloro-3,4-dihydroxy- benzoyl)amino]ethoxyimino methyl]-8-oxo-5-thia-1-azabicyclo [4.2.0]oct-2-ene-2-carboxylic acid in the form of hydrochloride .sup.1H-NMR (400 MHz, MeOH-d.sub.4, δ, ppm): 8.38 (s, 1H, azomethine), 6.83 (d, 1H, arom. H-6, J = 8.3 Hz), 6.74 (d, 1H, arom. H-5, J = 8.3 Hz), 5.91 (d, 1H, H-6, J = 5.0 Hz), 5.20 (d, 1H, H-7, J = 5.0 Hz), 4.24 (t, 2H, O—CH.sub.2, J = 5.4 Hz), 3.95 (d, 1H, H-2a, J = 17.9 Hz), 3.61 (t, 2H, N—CH.sub.2, J = 5.4 Hz), 3.53 (d, 1H, H-2b, J = 17.9 Hz), 1.63 (s, 3H, CH.sub.3), 1.62 (s, 3H, CH.sub.3). MS m/e: 713 [M + H].sup.+ [00015] N C(CH.sub.3).sub.2COOH 6 7-[[(2Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)-2-(fluoromethoxyimino)- acetyl]amino]-3-[2-[(2-chloro-3,4-dihydroxybenzoyl)amino)ethoxy- imino methyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxylic acid in the form of hydrochloride .sup.1H-NMR (400 MHz, MeOH-d.sub.4, δ, ppm): 8.42 (s, 1H, azomethine), 6.85 (d, 1H, arom. H-6, J = 8.2 Hz), 6.76 (m, 1H, arom. H-5, J = 8.2), 5.95 (d, 1H, H-6, J = 5.0 Hz), 5.81 (d, 2H, O—CH.sub.2—F, J = 54.6), 5.24 (d, 1H, H-7, J = 5.0 Hz), 4.28 (t, 2H, O—CH.sub.2, J = 5.4 Hz), 4.03 (d, 1H, H-2a, J = 18.1 Hz), 3.64 (t, 2H, N—CH.sub.2, J = 5.4 Hz), 3.59 (d, 1H, H-2b, J = 18.2 Hz). MS m/e: 657 [M − H].sup.− [00016] N CH.sub.2F 7 7-[[(2Z)-2-(2-aminothiazol-4-yl)-2-(1-carboxy-1-methylethoxy) imino-acetyl]amino]-3-[[3-[(2-chloro-3,4-dihydroxy-benzoyl) amino]-transcyclobutoxy]iminomethyl]-8-oxo-5-thia-1-azabicyclo [4.2.0]oct-2-ene-2-carboxylic acid in the form of a hydrochloride .sup.1H-NMR (400 MHz, MeOH-d.sub.4, δ, ppm, characteristic signals): 8.43 (s, 1H, azomethine), 6.89 (s, 1H, thiazole-H), 6.82 (d, 1H, arom. H-6, J = 8.3 Hz), 6.75 (d, 1H, arom. H-5, J = 8.3 Hz), 5.95 (d, 1H, H-6, J = 5.0 Hz), 5.24 (d, 1H, H-7, J = 5.0 Hz), 4.53 (m, 1H, cyclobutane), 4.01 (d, 1H, H-2a, J = 18.0 Hz), 3.61 (d, 1H, H-2b, [00017] CH C(CH.sub.3).sub.2COOH J = 18.0 Hz), 2.57 (m, 2H, cyclobutane), 2.40 (m, 2H, cyclobutane), 1.62 (s, 3H, CH.sub.3), 1.60 (s, 3H, CH.sub.3). MS m/e: 738 [M + H].sup.+ 8 7-[[(2Z)-2-(2-aminothiazol-4-yl)-2-(1-carboxy-1-methylethoxy) imino-acetyl]amino]-3-{[(3S)-1-(2-chloro-3,4-dihydroxybenzoyl) pyrrolidine-3-yl]oxyiminomethyl}-8-oxo-5-thia-1-azabicyclo[4.2.0] oct-2-ene-2-carboxylic acid in the form of hydrochloride .sup.1H-NMR (400 MHz, DMSO-d.sub.6, δ, ppm, characteristic signals): 8.27 (d, 1H, azomethine), 6.74 (m, 2H, arom. H-6, thiazole-H), 6.58 (dd, 1H, arom. H-5, J = 18.8, 8.4 Hz), 5.84 (m, 1H, H-6), 5.21 (m, 1H, H-7), 4.83-4.71 (2s, 1H, pyrrolidine), 3.90-3.10 (m, 6H, H-2, pyrrolidine), 2.10 (m, 2H, pyrrolidine), 1.44 (m, 6H, CH.sub.3). MS m/e: 738 [M + H].sup.+ [00018] CH C(CH.sub.3).sub.2COOH

[0123] Appropriate starting materials C2 and C3 (WO98/43981) are prepared according, e.g. analogously, to a method as conventional or according, e.g. analogously as set out in Example 1 Step 1

##STR00019##

[0124] Appropriate starting materials D2, D3 and D4 are prepared according, e.g. analogously, to a method as conventional or as set out below:

Intermediate D2

[0125] ##STR00020##

[0126] To a solution of 2-chloro-3,4-bis[(4-methoxyphenyl)methoxy]benzoic acid (EP 0 416 410) (1 g, 2.33 mmol) in dichloromethane was added HOBT (316 mg, 1 eq), EDC (446 mg, 1 eq) and O-(2-aminoethyl)hydroxylamine hydrochloride (348 mg, 1 eq) and stirred for 1 h at room temperature. Then triethylamine (976 μL, 3 eq.) was added and the resulting reaction mixture was stirred at room temperature for further 2 h. The resulting reaction mixture was concentrated to dryness and the residue was purified by chromatography (ethyl acetate/methanol=10/1) to give compound D2 as white solid (1.92 g).

[0127] .sup.1H-NMR (400 MHz, MeOH-d.sub.4, δ, ppm): 7.41 (m, 2H, arom.), 7.30 (m, 2H, arom.), 7.22 (d, 1H, arom., J=8.5 Hz), 7.12 (d, 1H, arom., J=8.5 Hz), 6.96 (m, 2H, arom.), 6.82 (m, 2H, arom.), 5.11 (s, 2H, OCH.sub.2), 4.94 (s, 2H, OCH.sub.2), 3.83 (s, 3H, OCH.sub.3), 3.80 (t, NOCH.sub.2, J=5.3 Hz), 3.79 (s, 3H, OCH.sub.3), 3.58 (t, 2H, NCH.sub.2, J=5.3 Hz).

[0128] MS m/e: 487 [M+H].sup.+, 531 [M+HCOO.sup.−]

Intermediate D3

[0129] ##STR00021##

Step a

[0130] To a solution of N-hydroxy-phthalimide (730 mg, 4.48 mmol) in DMSO was added potassium carbonate (1.55 g, 2.5 eq) and cis-tert-butyl N-(3-iodocyclobutyl)carbamate (1.99 g, 1.5 eq). The resulting reaction mixture was stirred for 3 h at 80° C. and then added slowly to ice water. The resulting precipitate was filtered, dried under reduced pressure and purified by chromatography (dichloromethane/methanol=100/1) to give trans-tert-butyl N-[3-(1,3-dioxoisoindolin-2-yl)oxycyclobutyl]carbamate as white solid (290 mg).

[0131] MS m/e: 350 [M+NH.sub.4.sup.+]

Step b

[0132] To a solution of trans-tert-butyl N-[3-(1,3-dioxoisoindolin-2-yl)oxycyclobutyl]carbamate (2.26 g, 6.80 mmol) in dichloromethane was added hydrogen chloride (2M in diethylether, 12 mL) and stirred at room temperature overnight. The resulting reaction mixture was concentrated to dryness to give trans-2-(3-aminocyclobutoxy)isoindoline-1,3-dione as light-brown solid (1.84 g wet).

[0133] MS m/e: 233 [M+H].sup.+

Step c

[0134] To a solution of 2-chloro-3,4-bis[(4-methoxyphenyl)methoxy]benzoic acid (EP 0 416 410) (3.40 g, 7.92 mmol) in dichloromethane was added HOBT (1.07 g, 1 eq), EDC (1.52 g, 1 eq) and trans-2-(3-aminocyclobutoxy)isoindoline-1,3-dione as light-brown solid (1.84 g wet, 6.80 mmol) and stirred for 1 h at room temperature. Then triethylamine (3.29 mL, 3 eq.) was added and the resulting reaction mixture was stirred at room temperature overnight. The resulting reaction mixture was concentrated to dryness and the residue was purified by chromatography (dichloromethane/methanol/ammonia=100/5/0.5) to give trans-2-chloro-N-[3-(1,3-dioxoisoindolin-2-yl)oxycyclobutyl]-3,4-bis[(4-methoxyphenyl)methoxy]benzamide as a light-yellow solid (2.49 g).

Step d

[0135] To a solution of trans-2-chloro-N-[3-(1,3-dioxoisoindolin-2-y0oxycyclobutyl]-3,4-bis[(4-methoxyphenyl)methoxy]benzamide (1.99 g, 3.09 mmol) in acetonitrile was added hydrazine hydrate (64-65% hydrazine, 0.30 mL, 2 eq) and stirred for 4 h at room temperature. The resulting reaction mixture was filtered. The filtrate was diluted with aqueous ammonium solution and extracted twice with ethyl acetate. The organic phases were combined, dried over Na.sub.2SO.sub.4, and concentrated to give compound D3 as white solid (0.98 g).

[0136] MS m/e: 513 [M+H].sup.+

Intermediate D4

[0137] ##STR00022##

Step a

[0138] To a solution of (3R)-hydroxypyrrolidine (919 mg, 10.56 mmol) in dichloromethane was added 2-chloro-3,4-bis[(4-methoxyphenyl)methoxy]benzoic acid (EP 0 416 410) (4.53 g, 1 eq), EDC (2.02 g, 1 eq) and HOBT (1.43 g, 1 eq) and stirred at room temperature overnight. The resulting reaction mixture was diluted with dichloromethane and washed with water. The resulting organic phase was concentrated to dryness and purified by chromatography (dichloromethane/methanol/ammonia=100/5/0.5) to give [2-chloro-3,4-bis[(4-methoxyphenyl)methoxy]phenyl]-[(3R)-3-hydroxypyrrolidin-1-yl]methanone (3.93 g).

[0139] MS m/e: 498 [M+H].sup.+

Step b

[0140] To a solution of [2-chloro-3,4-bis[(4-methoxyphenyl)methoxy]phenyl]-[(3R)-3-hydroxypyrrolidin-1-yl]methanone (3.77 g, 7.57 mmol) in THF was added triethylamine (1.59 mL, 1.5 eq) and methanesulfonyl chloride (0.71 mL, 1.2 eq) under ice cooling and stirred at room temperature for 2 days. The resulting reaction mixture was concentrated, distributed between ethyl acetate and water, and the resulting organic phase washed with saturated aqueous NaCl solution. The organic phase was again separated, dried over Na.sub.2SO.sub.4, and evaporated to dryness to give crude [(3R)-1-[2-chloro-3,4-bis [(4-methoxyphenyl)methoxy]benzoyl]pyrrolidin-3-yl] methanesulfonate (4.36 g wet).

[0141] MS m/e: 573 [M+H].sup.+

Step c

[0142] To a solution of crude [(3R)-1-[2-chloro-3,4-bis[(4-methoxyphenyl)methoxy]benzoyl]pyrrolidin-3-yl] methanesulfonate (3.83 g, 6.65 mmol) in DMSO was added potassium carbonate (2.30 g, 2.5 eq) and N-hydroxy-phthalimide (2.00 g, 1 eq) and stirred for 3 h at 80° C. The reaction mixture was distributed between ethyl acetate and water, and the resulting organic phase washed with saturated aqueous NaCl solution. The organic phase was again separated, dried over Na.sub.2SO.sub.4, and was concentrated to dryness to give crude 2-[(3S)-1-[2-chloro-3,4-bis[(4-methoxyphenyl)methoxy]benzoyl]pyrrolidin-3-yl]oxyisoindoline-1,3-dione (2.73 g wet).

[0143] MS m/e: 643 [M+H].sup.+

Step d

[0144] To a solution of crude 2-[(3S)-1-[2-chloro-3,4-bis[(4-methoxyphenyl)methoxy]benzoyl]pyrrolidin-3-yl]oxyisoindoline-1,3-dione (2.96 g, 4.60 mmol) in acetonitrile (20 mL) was added hydrazine hydrate (64-65% hydrazine, 0.45 mL, 2 eq) and stirred for 1 h at room temperature. The resulting reaction mixture was diluted with aqueous ammonium solution and extracted twice with ethyl acetate. The organic phases were combined, dried over Na.sub.2SO.sub.4, and concentrated to give crude [(3S)-3-aminooxypyrrolidin-1-yl]-[2-chloro-3,4-bis[(4-methoxyphenyl)methoxy]phenyl]methanone (1.83 g wet).

[0145] MS m/e: 513 [M+H].sup.+

Step e

[0146] To a solution of crude [(3S)-3-aminooxypyrrolidin-1-yl]-[2-chloro-3,4-bis[(4-methoxyphenyl)methoxy]phenyl]methanone (1.01 g, 1.97 mmol) in dichloromethane (10 mL) was added trifluoroacetic acid (5 mL) and stirred for 1 h at room temperature. The resulting reaction mixture was concentrated and hydrogen chloride (2M in diethylether, 1 eq) was added and again evaporated to dryness to give crude (3S)-3-aminooxypyrrolidin-1-yl]-(2-chloro-3,4-dihydroxy-phenyl)methanone hydrochloride (1.36 g).

[0147] MS m/e: 273 [MH].sup.+