Heterocyclic compounds and their use in preventing or treating bacterial infections

Abstract

The present invention relates to heterocyclic compounds, their process of preparation, pharmaceutical compositions comprising these compounds and use thereof, optionally in combination with other antibacterial agents and/or beta-lactam compounds, for the prevention or treatment of bacterial infections. The present invention also relates to the use of these compounds as -lactamase inhibitors and/or as antibacterial agents.

Claims

1. A compound of formula (I) ##STR00043## wherein: A-B represents: a CHCH double bond, wherein A is one of the carbon atoms of the CHCH double bond and B is the other carbon atom of the CHCH double bond; or a ring W, unsubstituted or substituted by one or more T and having carbon atoms at the positions corresponding to the positions of A and B, which represents one of the following: a phenyl group; a 5-membered aromatic heterocycle comprising one, two, or three heteroatom(s) independently selected from the group consisting of O, N, N(R.sup.3), S, S(O) or S(O).sub.2; or a 6-membered aromatic heterocycle comprising 1, 2, of 3 nitrogen heteroatom(s); R.sup.1 represents oxazole, isoxazole, thiazole, isothiazole, pyrazole, imidazole, imidazoline, oxadiazole, thiadiazole, triazole, tetrazole, unsubstituted or substituted by one or more T.sup.1; R.sup.2 represents SO.sub.3H, CFHCO.sub.2H or CF.sub.2CO.sub.2H; R.sup.3 independently represents hydrogen; (CH.sub.2).sub.mCN; (CH.sub.2).sub.nOC(O)Q.sup.1; (CH.sub.2).sub.mC(O)OQ.sup.1; (CH.sub.2).sub.nOC(O)OQ.sup.1; (CH.sub.2).sub.nOC(O)NQ.sup.1Q.sup.2; (CH.sub.2).sub.mC(O)NQ.sup.1Q.sup.2; (CH.sub.2).sub.mC(O)NQ.sup.1OQ.sup.2; (CH.sub.2).sub.mC(O)NQ.sup.1-NQ.sup.1Q.sup.2; (CH.sub.2).sub.nNQ.sup.1C(O)Q.sup.2; (CH.sub.2).sub.nNQ.sup.1C(O)OQ.sup.2; (CH.sub.2).sub.nNQ.sup.1C(O)NQ.sup.1Q.sup.2; (CH.sub.2).sub.nOQ.sup.1; (CH.sub.2).sub.nNQ.sup.1Q.sup.2; (CH.sub.2).sub.nNHC(NHQ.sup.3)=NQ.sup.4; (CH.sub.2).sub.nNHCHNQ.sup.3; (CH.sub.2).sub.mC(NHQ.sup.3)=NQ.sup.4; (Y)C.sub.1-C.sub.6-alkyl; (Y)C.sub.1-C.sub.6-fluoroalkyl; (Y)C.sub.3-C.sub.6-cycloalkyl; (Y)C.sub.3-C.sub.6-cyclofluoroalkyl; (Y)(CH.sub.2).sub.pCN; (Y)(CH.sub.2).sub.pOC(O)Q.sup.1; (Y)(CH.sub.2).sub.pC(O)OQ.sup.1; (Y)(CH.sub.2).sub.pOC(O)OQ.sup.1; (Y)(CH.sub.2).sub.pOC(O)NQ.sup.1Q.sup.2; (Y)(CH.sub.2).sub.pC(O)NQ.sup.1Q.sup.2; (Y)(CH.sub.2).sub.pC(O)NQ.sup.1OQ.sup.2; (Y)(CH.sub.2).sub.pC(O)NQ.sup.1-NQ.sup.1Q.sup.2; (Y)(CH.sub.2).sub.pNQ.sup.1C(O)Q.sup.2; (Y)(CH.sub.2).sub.pNQ.sup.1C(O)OQ.sup.2; (Y)(CH.sub.2).sub.pNQ.sup.1C(O)NQ.sup.1Q.sup.2; (CH.sub.2).sub.nOQ.sup.1; (Y)(CH.sub.2).sub.pNQ.sup.1Q.sup.2; (Y)(CH.sub.2).sub.pNHC(NHQ.sup.3)=NQ.sup.4; (Y)(CH.sub.2).sub.pNHCHNQ.sup.3; (Y)(CH.sub.2).sub.pC(NHQ.sup.3)=NQ.sup.4; or R.sup.3, unsubstituted or substituted by one or more T.sup.2, independently represents-C.sub.1-C.sub.6-alkyl; C.sub.1-C.sub.6-fluoroalkyl; C.sub.3-C.sub.6-cycloalkyl; C.sub.3-C.sub.6-cyclofluoroalkyl; (CH.sub.2).sub.m-(aromatic, totally or partially unsaturated, 4- to 6-membered heterocycle comprising at least one nitrogen heteroatom) or (Y)(CH.sub.2).sub.m-(aromatic, totally or partially unsaturated, 4- to 6-membered heterocycle comprising at least one nitrogen heteroatom); T and T.sup.1, identical or different, independently represent a fluorine atom; OC.sub.1-C.sub.6-fluoroalkyl; (CH.sub.2).sub.mC.sub.3-C.sub.6-cyclofluoroalkyl; (CH.sub.2).sub.mCN; (CH.sub.2).sub.mOC(O)Q.sup.1; (CH.sub.2).sub.mC(O)OQ.sup.1; (CH.sub.2).sub.mOC(O)OQ.sup.1; (CH.sub.2).sub.mOC(O)NQ.sup.1Q.sup.2; (CH.sub.2).sub.mC(O)NQ.sup.1Q.sup.2; (CH.sub.2).sub.mC(O)NQ.sup.1OQ.sup.2; (CH.sub.2).sub.mC(O)NQ.sup.1-NQ.sup.1Q.sup.2; (CH.sub.2).sub.mNQ.sup.1C(O)Q.sup.2; (CH.sub.2).sub.mNQ.sup.1C(O)OQ.sup.2; (CH.sub.2).sub.mNQ.sup.1C(O)NQ.sup.1Q.sup.2; (CH.sub.2).sub.mOQ.sup.1; (CH.sub.2).sub.mNQ.sup.1Q.sup.2; (CH.sub.2).sub.mNHC(NHQ.sup.3)NQ.sup.4; (CH.sub.2).sub.mNHCHNQ.sup.3; (CH.sub.2).sub.mC(NHQ.sup.3)NQ.sup.4; -(L)-(CH.sub.2).sub.pCN; -(L)-(CH.sub.2).sub.nOC(O)Q.sup.1; -(L)-(CH.sub.2).sub.pC(O)OQ.sup.1; -(L)-(CH.sub.2).sub.nOC(O)OQ.sup.1; -(L)-(CH.sub.2).sub.nOC(O)NQ.sup.1Q.sup.2; -(L)-(CH.sub.2).sub.pC(O)NQ.sup.1Q.sup.2; -(L)-(CH.sub.2).sub.pC(O)NQ.sup.1OQ.sup.2; -(L)-(CH.sub.2).sub.pC(O)NQ.sup.1-NQ.sup.1Q.sup.2; -(L)-(CH.sub.2).sub.nNQ.sup.1C(O)Q.sup.2; -(L)-(CH.sub.2).sub.nNQ.sup.1C(O)OQ.sup.2; -(L)-(CH.sub.2).sub.nNQ.sup.1C(O)NQ.sup.1Q.sup.2; -(L)-(CH.sub.2).sub.nOQ.sup.1; -(L)-(CH.sub.2).sub.nNQ.sup.1Q.sup.2; -(L)-(CH.sub.2).sub.nNHC(NHQ.sup.3)NQ.sup.4; -(L)-(CH.sub.2).sub.nNHCHNQ.sup.3; -(L)-(CH.sub.2).sub.nC(NHQ.sup.3)NQ.sup.4; or T and T.sup.1, identical or different, unsubstituted or substituted by one or more T.sup.3, independently represent C.sub.1-C.sub.6-alkyl; C.sub.1-C.sub.6-fluoroalkyl; (CH.sub.2).sub.mC.sub.3-C.sub.6-cycloalkyl; -(L)-C.sub.1-C.sub.6-alkyl; -(L)-C.sub.1-C.sub.6-fluoroalkyl; -(L)-C.sub.3-C.sub.6-cycloalkyl; -(L)-C.sub.3-C.sub.6-cyclofluoroalkyl; (CH.sub.2).sub.m-(aromatic, totally or partially unsaturated, 4- to 6-membered heterocycle comprising one, two, or three nitrogen heteroatoms(s); -(L)-(CH.sub.2).sub.p-(aromatic, totally or partially unsaturated, 4- to 6-membered heterocycle comprising one, two, or three nitrogen heteroatoms(s); Q.sup.1 and Q.sup.2, identical or different, independently represent a hydrogen atom; (CH.sub.2).sub.nNHQ.sup.3; (CH.sub.2).sub.nNHC(NHQ.sup.3)NQ.sup.4; (CH.sub.2).sub.nNHCHNQ.sup.3; (CH.sub.2).sub.pC(NHQ.sup.3)NQ.sup.4; (CH.sub.2).sub.nOQ.sup.3; (CH.sub.2).sub.pCONHQ.sup.3; (CH.sub.2).sub.nNHCONHQ.sup.3 or Q.sup.1 and Q.sup.2, identical or different, unsubstituted or substituted by one or more T.sup.3, independently represent a C.sub.1-C.sub.3-alkyl; (CH.sub.2).sub.m-(aromatic, totally or partially unsaturated, 4-, 5- or 6-membered heterocycle comprising one, two, or three nitrogen heteroatoms(s); or Q.sup.1, Q.sup.2 and the nitrogen atom to which they are bonded, form an unsubstituted or substituted by one or more T.sup.3, aromatic, saturated or partially unsaturated, 4-, 5- or 6-membered heterocycle comprising one nitrogen heteroatom and optionally comprising 1, 2, or 3 further nitrogen heteroatoms; Q.sup.3 and Q.sup.4, identical or different, independently represent a hydrogen atom or a C.sub.1-C.sub.3-alkyl; T.sup.2 and T.sup.3, identical or different, independently represent a fluorine atom; C.sub.1-C.sub.6-alkyl; C.sub.1-C.sub.6-fluoroalkyl; OC.sub.1-C.sub.6-fluoroalkyl; (CH.sub.2).sub.mC.sub.3-C.sub.6-cycloalkyl; (CH.sub.2).sub.mC.sub.3-C.sub.6-cyclofluoroalkyl; (CH.sub.2).sub.mCN; (CH.sub.2).sub.mOC(O)Q.sup.3; (CH.sub.2).sub.mC(O)OQ.sup.3; (CH.sub.2).sub.mOC(O)OQ.sup.3; (CH.sub.2).sub.mOC(O)NQ.sup.3Q.sup.4; (CH.sub.2).sub.mC(O)NQ.sup.3Q.sup.4; (CH.sub.2).sub.mC(O)NQ.sup.3OQ.sup.4; (CH.sub.2).sub.mC(O)NQ.sup.3-NQ.sup.3Q.sup.4; (CH.sub.2).sub.mNQ.sup.3C(O)Q.sup.4; (CH.sub.2).sub.mNQ.sup.3C(O)OQ.sup.4; (CH.sub.2).sub.mNQ.sup.3C(O)NQ.sup.3Q.sup.4; (CH.sub.2).sub.mOQ.sup.1; (CH.sub.2).sub.mNQ.sup.3Q.sup.4; (CH.sub.2).sub.mNHC(NHQ.sup.3)NQ.sup.4; (CH.sub.2).sub.mNHCHNQ.sup.3; (CH.sub.2).sub.mC(NHQ.sup.3)NQ.sup.4; -(L)-(CH.sub.2).sub.pC.sub.3-C.sub.6-cycloalkyl; -(L)-(CH.sub.2).sub.pC.sub.3-C.sub.6-cyclofluoroalkyl; -(L)-(CH.sub.2).sub.pCN; -(L)-(CH.sub.2).sub.nOC(O)Q.sup.3; -(L)-(CH.sub.2).sub.pC(O)OQ.sup.3; -(L)-(CH.sub.2).sub.nOC(O)OQ.sup.3; -(L)-(CH.sub.2).sub.nOC(O)NQ.sup.3Q.sup.4; -(L)-(CH.sub.2).sub.pC(O)NQ.sup.3Q.sup.4; -(L)-(CH.sub.2).sub.pC(O)NQ.sup.3OQ.sup.4; -(L)-(CH.sub.2).sub.pC(O)NQ.sup.3-NQ.sup.3Q.sup.4; -(L)-(CH.sub.2).sub.nNQ.sup.3C(O)Q.sup.4; -(L)-(CH.sub.2).sub.nNQ.sup.3C(O)OQ.sup.4; -(L)-(CH.sub.2).sub.nNQ.sup.3C(O)NQ.sup.3Q.sup.4; -(L)-(CH.sub.2).sub.nOQ.sup.1; -(L)-(CH.sub.2).sub.nNQ.sup.3Q.sup.4; -(L)-(CH.sub.2).sub.nNHC(NHQ.sup.3)NQ.sup.4; -(L)-(CH.sub.2).sub.nNHCHNQ.sup.3; -(L)-(CH.sub.2).sub.nC(NHQ.sup.3)NQ.sup.4; Y, identical or different, independently represents CO or S(O).sub.2; L, identical or different, independently represents O, S, N(R.sup.3), S(O) or S(O).sub.2; m independently represents 0, 1, 2, 3 or 4; n independently represents 2, 3 or 4; p independently represents 1, 2, 3 or 4 when Y is CO or 2, 3 or 4 when Y is S(O).sub.2; wherein: any carbon atom present within a group selected from alkyl, cycloalkyl, fluoroalkyl, cyclofluoroalkyl and heterocycle can be oxidized to form a CO group; any sulphur atom present within a heterocycle can be oxidized to form a SO group or a S(O).sub.2 group; and any nitrogen atom present within a heterocycle or present within a group that is trisubstituted thus forming a tertiary amino group can be further quaternized by a methyl group such that the quaternized amino group in conjunction with a counter ion maintains the pharmaceutical acceptability of the pharmaceutical compound; or a pharmaceutically acceptable salt, a zwitterion, an optical isomer, a racemate, a diastereoisomer, an enantiomer, a geometric isomer or a tautomer thereof.

2. The compound according to claim 1 that is of formula (I*) ##STR00044##

3. The compound according to claim 1 that is selected from the group of formula (A), (A*), (B), and (B*) ##STR00045##

4. The compound according to claim 1 selected from the group of compounds consisting of formulae (A1) to (A26) ##STR00046## ##STR00047## ##STR00048## ##STR00049## wherein X represents O, N(R.sup.3), S, S(O), or S(O).sub.2.

5. The compound according to claim 1 selected from the group consisting of compounds of formulae (A1*) to (A26*) ##STR00050## ##STR00051## ##STR00052## ##STR00053## wherein X represents O, N(R.sup.3), S, S(O), or S(O).sub.2.

6. The compound according to claim 1 selected from the group consisting of compounds of formula A1, A4, A12 to A23, A*, A4*, and A12* to A23* ##STR00054## ##STR00055## ##STR00056## ##STR00057## wherein X represents O N(R.sup.3), S, S(O), or S(O).sub.2.

7. A pharmaceutical composition comprising at least one compound according to claim 1 at an effective amount, and pharmaceutically acceptable carrier.

8. The pharmaceutical composition according to claim 7 further comprising one or more antibacterial compounds.

9. The pharmaceutical composition according to claim 7 further comprising one or more -lactam compounds.

10. The pharmaceutical composition according to claim 7 further comprising-one or more antibacterial compounds and one or more -lactam compounds.

11. The pharmaceutical composition according to claim 10 wherein: the one or more antibacterial compounds are selected from aminoglycosides, -lactams, glycylcyclines, tetracyclines, quinolones, fluoroquinolones, glycopeptides, lipopeptides, macrolides, ketolides, lincosamides, streptogramins, oxazolidinones, polymyxins and mixtures thereof; and the one or more -lactam compounds are selected from -lactams and mixtures thereof.

12. A kit comprising at least two pharmaceutical compositions according to claim 7.

13. A method for treating a bacterial infection, the method comprising B administering, to a person in need thereof, the compound according to claim 1.

14. The method according to claim 13, wherein the bacterial infection is caused by bacteria producing one or more -lactamase, a gram-positive bacteria, or a gram-negative bacteria.

15. A method for treating a bacterial infection, the method comprising administering, to a person in need thereof, the composition according to claim 11.

16. The method according to claim 15, wherein the bacterial infection is caused by bacteria producing one or more -lactamase.

17. The method according to claim 15, wherein the bacterial infection is caused by a gram-positive bacteria or by gram-negative bacteria.

18. A method for treating bacterial infections, the method comprising simultaneously, separately, or sequentially administering, to a person in need thereof, the least two pharmaceutical compositions of the kit according to claim 12.

Description

EXAMPLES

(1) Abbreviations or symbols used herein include: ACHN: 1,1-azobis(cyclohexanecarbonitrile) ACN: acetonitrile AcCl: acetyl chloride AcOH: acetic acid Bn: benzyl BOC: tert-butoxycarbonyl Boc.sub.2O: tert-butoxycarbonyl anhydride bs: broad singlet CFU: colony-forming units CLSI: clinical laboratory standards institute d: doublet DBU: 1,8-diazabicyclo[5.4.0]undec-7-ene DCM: dichloromethane dd: doublet of doublet ddd: doublet of doublet of doublet ddt: doublet of doublet of triplet dt: doublet of triplet DTA: di-tert-butylazodicarboxylate DEAD: diethyl azodicarboxylate DIAD: diisopropyl azodicarboxylate DIPEA: diisopropylethylamine DMF: N,N-dimethylformamide DMAP: 4-dimethylaminopyridine DMSO: dimethylsulfoxide EtOAc: ethyl acetate Et.sub.2O: diethyl ether h: hours HATU: 1-[Bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxid hexafluorophosphate IC.sub.50: concentration of inhibitor responsible for 50% of inhibition KOAc: potassium acetate m: massif min: minutes MeOH: methanol MIC: minimum inhibitory concentration MS: mass spectrometry MsCl: methanesulfonyl chloride MTBE: methyl tert-butyl ether NBS: N-bromosuccinimide NMR: nuclear magnetic resonance spectroscopy Nos: nosyl, nitrobenzenesulfonyl Pd(dppf)Cl.sub.2: [1,1-bis(diphenylphosphino)ferrocene]dichloropalladium Pd(PPh.sub.3).sub.4: tetrakis(triphenylphosphine)palladium(0) PEPPSI: [1,3-bis(2,6-diisopropylphenyl)imidazol-2-ylidene](3-chloropyridyl)palladium(II) dichloride PG1: protective group 1 PG2: protective group 2 PhSH: thiophenol PivCl: pivaloyl chloride PPh.sub.3: triphenylphosphine Ppm: parts per million q: quartet dq: doublet of quartet rt: room temperature s: singlet SEMCl: 2-(trimethylsilyl)ethoxymethyl chloride t: triplet td: triplet of doublet TEA: trimethylamine TFA: trifluoroacetic acid THF: tetrahydrofuran TLC: thin layer chromatography TMSCN: trimethylsilyl cyanide TMSI: iodotrimethylsilane

(2) The compounds of the present invention of formula (I) can be prepared by the following reaction Schemes 1-9 where A-B represents a ring W or CHCH double bond.

(3) ##STR00015## ##STR00016##

(4) ##STR00017##

(5) ##STR00018## ##STR00019##

(6) ##STR00020## ##STR00021##

(7) ##STR00022## ##STR00023##

(8) ##STR00024##

(9) ##STR00025## ##STR00026##

(10) ##STR00027##

(11) ##STR00028## ##STR00029##

(12) ##STR00030## ##STR00031##

(13) ##STR00032## ##STR00033##

(14) ##STR00034##

Example 1: Synthesis of Sodium and 2,2,2-trifluoroacetate [trans-7-(1H-imidazol-3-ium-2-yl)-9-oxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.SUP.2,6.]undeca-2(6),4-dien-10-yl]sulfate

(15) ##STR00035## ##STR00036## ##STR00037## ##STR00038##

Step 1: Preparation of Intermediate tert-butyl 7-oxo-4,6-dihydrothiazolo[4,5-c]pyridine-5-carboxylate (1a)

(16) To a solution of tert-butyl 2-amino-7-oxo-4,6-dihydrothiazolo[4,5-c]pyridine-5-carboxylate (described in patent WO2002/100860) (15.87 g, 58.9 mmol) in anhydrous THF (500 mL) under inert atmosphere was added isopentyl nitrite (40 mL, 294 mmol) under 4 days. The solvent was evaporated in vacuo. The residue was solubilized in DCM and washed with water and brine. The organic layer was dried over Na.sub.2SO.sub.4 and evaporated in vacuo. The red solid was purified by flash chromatography on silica gel (DCM) to give tert-butyl 7-oxo-4,6-dihydrothiazolo[4,5-c]pyridine-5-carboxylate (1a) (10.83 g, 42.58 mmol, 72%) as an off-white solid.

(17) MS m/z ([M+H]) 255.

(18) .sup.1H NMR (400 MHz, CDCl.sub.3): (ppm) 1.49 (s, 9H), 4.35 (s, 2H), 4.95 (s, 2H), 9.08 (s, 1H).

Step 2: Preparation of Intermediate tert-butyl 7-hydroxy-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-5-carboxylate (1b)

(19) To a solution of tert-butyl 7-oxo-4,6-dihydrothiazolo[4,5-c]pyridine-5-carboxylate (1a) (27.2 g, 107 mmol) in MeOH (1550 mL) under inert atmosphere at 0 C. was portionwise added NaBH.sub.4 (4.53 g, 120 mmol). The reaction mixture was stirred for 1 h at 0 C., then hydrolyzed with water and concentrated in vacuo. The residue was solubilized with DCM, washed with water and brine. The organic layer was dried over Na.sub.2SO.sub.4, evaporated in vacuo to give tert-butyl 7-hydroxy-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-5-carboxylate (1b) (27.4 g, 106.9 mmol, quantitative yield) as a yellow solid without further purification.

(20) MS m/z ([M+H]) 257.

(21) .sup.1H NMR (400 MHz, CDCl.sub.3): (ppm) 1.49 (s, 9H), 3.69 (d, J=13.6 Hz, 1H), 3.97 (dd, J=13.6/4.6 Hz, 1H), 4.50 (d, J=17.1 Hz, 1H), 4.86 (d, J=14.5 Hz, 1H), 4.97 (bs, 1H), 8.75 (s, 1H).

Step 3: Preparation of Intermediate tert-butyl 7-[allyloxy-(2-nitrophenyl)sulfonyl-amino]-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-5-carboxylate (1c)

(22) To a solution of tert-butyl 7-hydroxy-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-5-carboxylate (1b) (49.4 g, 193 mmol) in toluene (2.14 L) under inert atmosphere was added N-allyloxy-2-nitro-benzenesulfonamide (49.84 g, 193 mmol), PPh.sub.3 (50.62 g, 193 mmol) and DTA (50.15 g, 218 mmol) portionwise. The reaction mixture was stirred for 24 h at rt. The precipitate was filtered off and the filtrate was concentrated in vacuo. The residue was purified by flash chromatography on silica gel (petroleum ether/acetone 100/0 to 60/40) to give tert-butyl 7-[allyloxy-(2-nitrophenyl)sulfonyl-amino]-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-5-carboxylate (1c) (80.0 g, 161 mmol, 83%) as a yellow oil.

(23) MS m/z ([M+H]) 497.

(24) .sup.1H NMR (400 MHz, CDCl.sub.3): (ppm) 1.40 (s, 9H), 3.50 (bs, 1H), 4.00-4.50 (m, 4H), 4.75-5.05 (m, 3H), 5.25-5.60 (m, 2H), 7.64 (d, J=8.0 Hz, 1H), 7.73 (ddd, 7.8/7.8/1.5 Hz, 1H), 7.82 (ddd, J=7.8/7.8/1.5 Hz, 1H), 8.09 (d, J=8.0 Hz, 1H), 8.71 (s, 1H).

Step 4: Preparation of Intermediate N-allyloxy-2-nitro-N-(4,5,6,7-tetrahydrothiazolo[4,5-c]pyridin-7-yl)benzenesulfonamide (1d)

(25) To a solution of tert-butyl 7-[allyloxy-(2-nitrophenyl)sulfonyl-amino]-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-5-carboxylate (1c) (20.0 g, 40.28 mmol) in anhydrous DCM (37 mL) under inert atmosphere at 0 C. was dropwisely added TFA (37 mL, 483 mmol). After stirring for 2 h at rt, the reaction mixture was cooled at 0 C. and basified with ammonium hydroxide solution 28% until pH 8. The mixture was diluted with water and extracted with DCM. The organic layer was dried over Na.sub.2SO.sub.4, concentrated in vacuo to give N-allyloxy-2-nitro-N-(4,5,6,7-tetrahydrothiazolo[4,5-c]pyridin-7-yl)benzenesulfonamide (1d) (14.7 g, 37.0 mmol, 92%) as a yellow solid without further purification.

(26) MS m/z ([M+H]) 397.

(27) .sup.1H NMR (400 MHz, CDCl.sub.3): (ppm) 1.73 (bs, 1H), 3.03 (dd, J=14.7/4.9 Hz, 1H), 3.19 (bs, 1H), 3.88 (d, J=17.1 Hz, 1H), 4.02 (bs, 1H), 4.10 (d, J=17.1 Hz, 1H), 4.37 (dd, J=11.4/6.2 Hz, 1H), 5.02-5.13 (m, 2H), 5.14-5.20 (m, 1H), 5.47-5.59 (m, 1H), 7.65 (dd, J=7.7/1.4 Hz, 1H), 7.75 (ddd, J=7.7/7.7/1.4 Hz, 1H), 7.84 (ddd, J=7.7/7.7/1.5 Hz, 1H), 8.14 (dd, J=7.7/1.5 Hz, 1H), 8.77 (s, 1H).

Step 5: Preparation of Intermediate N-allyloxy-N-(6,7-dihydrothiazolo[4,5-c]pyridin-7-yl)-2-nitro-benzenesulfonamide (1e)

(28) To a solution of N-allyloxy-2-nitro-N-(4,5,6,7-tetrahydrothiazolo[4,5-c]pyridin-7-yl)benzenesulfonamide (1d) (14.7 g, 37.0 mmol) in anhydrous DCM (225 mL) under inert atmosphere at 0 C. was dropwisely added a solution of N-chlorosuccinimide (6.43 g, 48.14 mmol) in DCM (225 mL). After stirring for 4 h at 0 C., DIPEA (24 mL, 138 mmol) was added and the reaction mixture was stirred for 18 h at rt. The reaction mixture was cooled at 0 C., washed with a tartaric acid solution (2 eq/DIPEA) and extracted with DCM. The organic layer was dried over Na.sub.2SO.sub.4 and concentrated in vacuo to give N-allyloxy-N-(6,7-dihydrothiazolo[4,5-c]pyridin-7-yl)-2-nitro-benzenesulfonamide (1e) (14.6 g, 37.0 mmol, quantitative yield) as an ochre solid without further purification.

(29) MS m/z ([M+H]) 395.

Step 6: Preparation of Intermediate N-allyloxy-N-(4-cyano-4,5,6,7-tetrahydrothiazolo[4,5-c]pyridin-7-yl)-2-nitro-benzenesulfonamide (1f)

(30) To a solution of N-allyloxy-N-(6,7-dihydrothiazolo[4,5-c]pyridin-7-yl)-2-nitro-benzenesulfonamide (1e) (14.6 g, 37.0 mmol) in anhydrous DCM (260 mL) under inert atmosphere at 0 C. was added TMSCN (47 mL, 377 mmol). The reaction mixture was stirred for 9 days at rt. The solvent was evaporated and the residue was purified by flash chromatography on silica gel (DCM/acetone 90/10) to give N-allyloxy-N-(4-cyano-4,5,6,7-tetrahydrothiazolo[4,5-c]pyridin-7-yl)-2-nitro-benzenesulfonamide (1f) (12.7 g, 30.0 mmol, 81%) as a beige solid.

(31) MS m/z ([M+H]) 422.

(32) .sup.1H NMR (400 MHz, CDCl.sub.3): (ppm) 2.33 (bs, 1H), 3.38 (bs, 2H), 4.04 (bs, 1H), 4.36 (dd, J=11.4/6.1 Hz, 1H), 4.98-5.21 (m, 4H), 5.44-5.54 (m, 1H), 7.68 (dd, J=7.8/1.3 Hz, 1H), 7.76 (ddd, J=7.7/7.7/1.3 Hz, 1H), 7.86 (ddd, J=7.8/7.8/1.4 Hz, 1H), 8.12 (dd, J=7.7/1.4 Hz, 1H), 8.83 (s, 1H).

Step 7: Preparation of Intermediate methyl 7-[allyloxy-(2-nitrophenyl)sulfonyl-amino]-4,5,6,7-tetrahydrothiazolo[4,5-c]pyridine-4-carboxylate (1q)

(33) In MeOH (72 mL) at 0 C. under inert atmosphere was added AcCl (30 mL, 422 mmol). After stirring for 2 h at 0 C. then for 30 min at rt, a solution of N-allyloxy-N-(4-cyano-4,5,6,7-tetrahydrothiazolo[4,5-c]pyridin-7-yl)-2-nitro-benzenesulfonamide (1f) (12.7 g, 30.0 mmol) in MeOH (36 mL) was added. The mixture was heated at 50 C. for 18 h. The solvent was evaporated. The residue was extracted with DCM and washed with a saturated solution of NaHCO.sub.3. The organic layer was dried over Na.sub.2SO.sub.4 and concentrated in vacuo. The brown oil was purified by flash chromatography on silica gel (DCM/acetone 100/0 to 80/20) to give methyl 7-[allyloxy-(2-nitrophenyl)sulfonyl-amino]-4,5,6,7-tetrahydrothiazolo[4,5-c]pyridine-4-carboxylate (1g) (10.8 g, 23.7 mmol, 78%) as a yellow solid.

(34) MS m/z ([M+H]) 455.

(35) .sup.1H NMR (400 MHz, CDCl.sub.3): (ppm) 3.20-3.57 (m, 2H), 3.48, 3.54, 3.79 and 3.88 (s, 3H), 4.05-4.16 (m, 1H), 4.34 and 4.43 (dd, J=11.1/6.6 Hz, 1H), 4.90 and 5.00 (s, 1H), 5.02-5.21 (m, 2H), 5.27-5.41 (m, 1H), 5.47-5.69 (m, 1H), 7.67 (t, J=7.7 Hz, 1H), 7.74-7.81 (m, 1H), 7.82-7.89 (m, 1H), 8.13-8.19 (m, 1H), 8.83 and 8.84 (s, 1H).

(36) (Mixture of Diastereoisomers)

Step 8: Preparation of Intermediate O5-benzyl O4-methyl 7-[allyloxy-(2-nitrophenyl)sulfonyl-amino]-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-4,5-dicarboxylate (1h)

(37) To a solution of methyl 7-[allyloxy-(2-nitrophenyl)sulfonyl-amino]-4,5,6,7-tetrahydrothiazolo[4,5-c]pyridine-4-carboxylate (1g) (10.8 g, 23.7 mmol) in anhydrous DCM (230 mL) under inert atmosphere at 78 C. were successively added DIPEA (8.3 mL, 47.4 mmol) and benzyl chloroformate (4.1 mL, 28.4 mmol). The mixture was stirred at rt for 2.5 h then water was added. The aqueous layer was extracted with DCM. The organic layers were combined, washed with a saturated solution of sodium bicarbonate, dried over Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (DCM/acetone 100/0 to 90/10) to give O5-benzyl O4-methyl 7-[allyloxy-(2-nitrophenyl)sulfonyl-amino]-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-4,5-dicarboxylate (1h) (13.1 g, 22.2 mmol, 93%) as a yellow solid.

(38) MS m/z ([M+H]) 589.

Step 9: Preparation of Intermediate O5-benzyl O4-methyl 7-(allyloxyamino)-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-4,5-dicarboxylate (1i)

(39) To a solution of O5-benzyl O4-methyl 7-[allyloxy-(2-nitrophenyl)sulfonyl-amino]-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-4,5-dicarboxylate (1h) (13.1 g, 22.2 mmol) in ACN (145 mL) under inert atmosphere at 0 C. were successively added PhSH (11.4 mL, 111 mmol) and potassium carbonate (23 g, 167 mmol). The mixture was stirred at rt for 18 h then filtered through a pad of Celite and rinsed with DCM. The filtrate was concentrated in vacuo. The residue was purified by flash chromatography on silica gel (heptane/EtOAc 100/0 to 20/80) to give O5-benzyl O4-methyl 7-(allyloxyamino)-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-4,5-dicarboxylate (1i) (7.87 g, 19.5 mmol, 87%) as an ochre oil.

(40) MS m/z ([M+H]) 404.

Step 10: Preparation of Intermediate O5-benzyl O4-methyl 7-[allyloxy(chlorocarbonyl)amino]-6,7-dihydro-4H-thiazolo(chlorocarbon[4,5-c]pyridine-4,5-dicarboxylate (1j)

(41) To a solution of O5-benzyl O4-methyl 7-(allyloxyamino)-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-4,5-dicarboxylate (1i) (7.87 g, 19.5 mmol) in ACN (365 mL) under inert atmosphere at 10 C. was added TEA (8.2 mL, 58.5 mmol). A solution of diphosgene (3.1 mL, 25.3 mmol) diluted in ACN (45 mL) was dropwisely added. After stirring for 10 min at 0 C., the solvent was evaporated and the residue was purified by flash chromatography on silica gel (heptane/EtOAc 100/0 to 50/50) to give O5-benzyl O4-methyl 7-[allyloxy(chlorocarbonyl)amino]-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-4,5-dicarboxylate (1j) (8.44 g, 18.1 mmol, 93%) as a yellow oil.

(42) MS m/z ([M/M+2]) 466/468.

Step 11: Preparation of Intermediate methyl trans-10-allyloxy-9-oxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.SUP.2,6.]undeca-2(6),4-diene-7-carboxylate (1k)

(43) To a solution of O5-benzyl O4-methyl 7-[allyloxy(chlorocarbonyl)amino]-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-4,5-dicarboxylate (1j) (7.40 g, 15.9 mmol) in anhydrous DCM (100 mL) under inert atmosphere at 0 C. was dropwise added methanesulfonic acid (26 mL, 397 mmol). After stirring for 2 h at rt, the reaction mixture was poured at 78 C. in a solution of TEA (110 mL, 794 mmol) in DCM (74 mL) and stirred for 30 min. The mixture was washed with water and extracted with DCM. The organic layer was dried over Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (DCM/EtOAc 100/0 to 85/15) to give methyl trans-10-allyloxy-9-oxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2,6]undeca-2(6),4-diene-7-carboxylate (1k) (4.50 g, 15.2 mmol, 96%) as an off-white solid.

(44) MS m/z ([M+H]) 296.

(45) .sup.1H NMR (300 MHz, CDCl.sub.3): (ppm) 3.60 (dd, J=11.6/2.8 Hz, 1H), 3.70 (dd, J=11.6/0.8 Hz, 1H), 3.81 (s, 3H), 4.32-4.46 (m, 2H), 4.65 (dd, J=2.8/0.8 Hz, 1H), 5.25-5.35 (m, 3H), 5.88-6.01 (m, 1H), 8.68 (s, 1H).

Step 12: Preparation of Intermediate trans-10-allyloxy-9-oxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.SUP.2,6.]undeca-2(6),4-diene-7-carboxylic Acid (1l)

(46) To a solution of methyl trans-10-allyloxy-9-oxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2,6]undeca-2(6),4-diene-7-carboxylate (1k) (1.60 g, 5.42 mmol) in a mixture of THF (30 mL) and water (19 mL) at 0 C. was dropwisely added a 1N solution of lithium hydroxide (5.4 mL, 5.42 mmol). After stirring for 24 h, the mixture was diluted with DCM and acidified with a 2N solution of HCl (6 mL) at 0 C. The product was extracted with DCM and the organic layer was dried over Na.sub.2SO.sub.4 before being concentrated in vacuo. The crude was triturated with Et.sub.2O to give trans-10-allyloxy-9-oxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2,6]undeca-2(6),4-diene-7-carboxylic acid (1l) (1.52 g, 5.42 mmol, quantitative yield) as a white solid without further purification.

(47) MS m/z ([M+H]) 282.

(48) .sup.1H NMR (400 MHz, CDCl.sub.3): (ppm) 3.59 (d, J=11.7 Hz, 1H), 3.72 (dd, J=11.7/2.8 Hz, 1H), 4.18 (bs, 1H), 4.38-4.50 (m, 1H), 4.67 (d, J=2.6 Hz, 1H), 5.30-5.40 (m, 2H), 5.41 (s, 1H), 5.94-6.05 (m, 1H), 8.89 (s, 1H).

Step 13: Preparation of Intermediate trans-10-allyloxy-N-(2-bromoethyl)-9-oxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.SUP.2,6.]undeca-2(6),4-diene-7-carboxamide (1m)

(49) To a solution of trans-10-allyloxy-9-oxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2,6]undeca-2(6),4-diene-7-carboxylic acid (1l) (1.40 g, 4.98 mmol) in anhydrous THF (50 mL) under inert atmosphere at 50 C. was added 4-methylmorpholine (3.3 mL, 29.9 mmol). After stirring for 15 min trimethylacetyl chloride (0.92 mL, 7.47 mmol) was added and the mixture was maintained at 50 C. for 1 h. 2-Bromoethylamine hydrobromide (3.06 g, 14.9 mmol) was portionwise added and temperature was raised to 30 C. for 2 h. The precipitate was filtered off and washed with EtOAc. The filtrate was concentrated in vacuo to give trans-10-allyloxy-N-(2-bromoethyl)-9-oxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2,6]undeca-2(6),4-diene-7-carboxamide (1m) (1.93 g, 4.98 mmol, quantitative yield) as a yellow oil which was used without further purification without further purification.

(50) MS m/z ([M/M+2]) 387/389.

(51) .sup.1H NMR (400 MHz, CDCl.sub.3): (ppm) 3.39 (dd, J=11.5/0.07 Hz, 1H), 3.44-3.53 (m, 2H), 3.62-3.74 (m, 2H), 3.76-3.84 (m, 1H), 4.38-4.49 (m, 1H), 4.66 (d, J=2.3 Hz, 1H), 5.25 (s, 1H), 5.29-5.41 (m, 2H), 5.94-6.04 (m, 1H), 7.43 (dd, J=5.9/5.9 Hz, 1H), 8.75 (s, 1H).

Step 14: Preparation of Intermediate trans-10-allyloxy-N-(2-azidoethyl)-9-oxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.SUP.2,6.]undeca-2(6),4-diene-7-carboxamide (1n)

(52) To a solution of trans-10-allyloxy-N-(2-bromoethyl)-9-oxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2,6]undeca-2(6),4-diene-7-carboxamide (1m) (1.93 g, 4.98 mmol) in anhydrous DMF (50 mL) under inert atmosphere were successively added sodium iodide (1.49 g, 9.96 mmol) and sodium azide (0.65 g, 9.96 mmol). The reaction mixture was stirred for 1 h at 40 C. then overnight at rt. The solvent was evaporated. DCM was added to the residue. The solids were filtered off and rinsed with DCM. The filtrate was concentrated in vacuo. The residue was purified by flash chromatography on silica gel (DCM/acetone 100/0 to 80/20) then triturated in Et.sub.2O to give trans-10-allyloxy-N-(2-azidoethyl)-9-oxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2,6]undeca-2(6),4-diene-7-carboxamide (1n) (1.15 g, 3.29 mmol, 66%) as a white solid.

(53) MS m/z ([M+H]) 350.

(54) .sup.1H NMR (400 MHz, CDCl.sub.3): (ppm) 3.39 (d, J=11.5 Hz, 1H), 3.42-3.54 (m, 4H), 3.69 (dd, J=11.5/2.9 Hz, 1H), 4.37-4.48 (m, 2H), 4.65 (d, J=2.7 Hz, 1H), 5.24 (s, 1H), 5.29-5.38 (m, 2H), 5.93-6.03 (m, 1H), 7.35 (d, J=6.1 Hz, 1H), 8.74 (s, 1H).

Step 15: Preparation of Intermediate tert-butyl N-(2-azidoethyl)-N-[trans-10-allyloxy-9-oxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.SUP.2,6.]undeca-2(6),4-diene-7-carbonyl)]carbamate (1o)

(55) To a solution of trans-10-allyloxy-N-(2-azidoethyl)-9-oxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2,6]undeca-2(6),4-diene-7-carboxamide (1n) (1.15 g, 3.29 mmol) in anhydrous ACN (33 mL) under inert atmosphere at rt were successively added Boc.sub.2O (1.08 g, 4.94 mmol) and 4-dimethylaminopyridine (40 mg, 0.33 mmol). The mixture was stirred for 3 days. The solvent was evaporated in vacuo and the residue was purified by flash chromatography on silica gel (DCM/Et.sub.2O 100/0 to 90/10) to give tert-butyl N-(2-azidoethyl)-N-[trans-10-allyloxy-9-oxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2,6]undeca-2(6),4-diene-7-carbonyl)]carbamate (10) (1.10 g, 2.45 mmol, 74%) as a yellow solid.

(56) MS m/z ([M+H]) 450.

(57) .sup.1H NMR (400 MHz, CDCl.sub.3): (ppm) 1.62 (s, 9H), 3.47 (dd, J=6.0 Hz, 2H), 3.52 (d, J=11.4 Hz, 1H), 3.59 (dd, J=11.5/2.8 Hz, 1H), 3.88 (ddd, J=14.0/6.4/6.4 Hz, 1H), 4.00 (ddd, J=14.0/5.8/5.8 Hz, 1H), 4.38-4.49 (m, 2H), 4.64 (d, J=2.8 Hz, 1H), 5.30-5.39 (m, 2H), 5.95-6.05 (m, 1H), 6.44 (s, 1H), 8.70 (s, 1H).

Step 16: Preparation of Intermediate tert-butyl 2-(trans-10-allyloxy-9-oxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.SUP.2,6.]undeca-2(6),4-dien-7-yl)-4,5-dihydroimidazole-1-carboxylate (1p)

(58) To a solution of tert-butyl N-(2-azidoethyl)-N-[trans-10-allyloxy-9-oxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2,6]undeca-2(6),4-diene-7-carbonyl)]carbamate (1o) (1.10 g, 2.45 mmol) in anhydrous toluene (41 mL) under inert atmosphere was added PPh.sub.3 (0.64 g, 2.45 mmol). The mixture was stirred for 16 h at rt, deposited on silica and purified by flash chromatography on silica gel (Heptane/Acetone 100/0 to 0/100) to give tert-butyl 2-(trans-10-allyloxy-9-oxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2,6]undeca-2(6),4-dien-7-yl)-4,5-dihydroimidazole-1-carboxylate (1p) (910 mg, 2.24 mmol, 91%) as a white solid.

(59) MS m/z ([M+H]) 406.

(60) .sup.1H NMR (400 MHz, CDCl.sub.3): (ppm) 1.56 (s, 9H), 3.42 (dd, J=11.3/2.9 Hz, 1H), 3.64-3.93 (m, 5H), 4.33-4.47 (m, 2H), 4.61 (d, J=2.2 Hz, 1H), 5.24-5.35 (m, 2H), 5.90-6.04 (m, 1H), 6.32 (s, 1H), 8.66 (s, 1H).

Step 17: Preparation of Intermediate trans-10-allyloxy-7-(4,5-dihydro-1H-imidazol-3-ium-2-yl)-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.SUP.2,6.]undeca-2(6),4-dien-9-one trifluoroacetate (1q)

(61) To a solution of tert-butyl 2-(trans-10-allyloxy-9-oxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2,6]undeca-2(6),4-dien-7-yl)-4,5-dihydroimidazole-1-carboxylate (1p) (240 mg, 0.59 mmol) in anhydrous DCM (4.3 mL) under inert atmosphere at 0 C. was dropwisely added TFA (4.3 mL, 56.2 mmol). The mixture was stirred at rt for 16 h then concentrated in vacuo. The residue was co-evaporated with toluene to give trans-10-allyloxy-7-(4,5-dihydro-1H-imidazol-3-ium-2-yl)-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2,6]undeca-2(6),4-dien-9-one trifluoroacetate (1q) (247 mg, 0.59 mmol, quantitative yield) without further purification.

(62) MS m/z ([M+H]) 306.

Step 18: Preparation of trans-10-allyloxy-7-(1H-imidazol-2-yl)-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.SUP.2,6.]undeca-2(6),4-dien-9-one (1r)

(63) To a solution of oxalyl chloride (0.1 mL, 1.18 mmol) in DCM (9 mL) under inert atmosphere at 65 C. was dropwise added dimethyl sulfoxide (0.17 mL, 2.36 mmol). The mixture was stirred for 30 min then a solution of trans-10-allyloxy-7-(4,5-dihydro-1H-imidazol-3-ium-2-yl)-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2,6]undeca-2(6),4-dien-9-one trifluoroacetate (1q) (247 mg, 0.59 mmol) in anhydrous DCM (9 mL) was dropwisely added. After 5 min at 65 C., TEA (0.82 mL, 5.89 mmol) was added and the temperature was raised to 15 C. for 3 h. The reaction mixture was quenched with water and the product was extracted with DCM. The brown oil was purified by flash chromatography on silica gel (heptane/acetone 100/0 to 0/100) to give trans-10-allyloxy-7-(1H-imidazol-2-yl)-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2,6]undeca-2(6),4-dien-9-one (1r) (59 mg, 0.19 mmol, 32%) as an off-white solid.

(64) MS m/z ([M+H]) 304.

(65) .sup.1H NMR (400 MHz, CDCl.sub.3): (ppm) 3.49 (dd, J=11.4/2.9 Hz, 1H), 3.76 (d, J=11.4 Hz, 1H), 4.40-4.52 (m, 2H), 4.69 (d, J=2.3 Hz, 1H), 5.31-5.41 (m, 2H), 5.86 (s, 1H), 5.96-6.06 (m, 1H), 7.04 (s, 2H), 8.74 (s, 1H), 10.48 (bs, 1H).

Step 19: Preparation of trans-10-allyloxy-7-[1-(2-trimethylsilylethoxymethyl)-1H-imidazol-2-yl]-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.SUP.2,6.]undeca-2(6),4-dien-9-one (1s)

(66) To a solution of trans-10-allyloxy-7-(1H-imidazol-2-yl)-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2,6]undeca-2(6),4-dien-9-one (1r) (59 mg, 0.19 mmol) in anhydrous DMF (2 mL) under inert atmosphere were successively added (2-(chloromethoxy)ethyl)trimethylsilane (0.10 mL, 0.58 mmol) and TEA (0.08 mL, 0.58 mmol). After stirring for 16 h at rt, the mixture was concentrated in vacuo. The residue was purified by flash chromatography on silica gel (DCM/Acetone 80/20 to 0/100) to give trans-10-allyloxy-7-[1-(2-trimethylsilylethoxymethyl)-1H-imidazol-2-yl]-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2,6]undeca-2(6),4-dien-9-one (1s) (43 mg, 0.10 mmol, 51%) as a yellow oil.

(67) MS m/z ([M+H]) 434.

(68) .sup.1H NMR (300 MHz, CDCl.sub.3): (ppm) 0.01 (s, 9H), 0.89-1.06 (m, 2H), 3.35 (dd, J=11.3/2.9 Hz, 1H), 3.64 (d, J=8.2 Hz, 2H), 3.78 (dd, J=11.3/0.7 Hz, 1H), 4.38-4.51 (m, 2H), 4.68 (d, J=2.2 Hz, 1H), 5.29-5.43 (m, 3H), 5.66 (d, J=10.7 Hz, 1H), 5.95 (s, 1H), 5.95-6.08 (m, 1H), 6.99 (d, J=1.3 Hz, 1H), 7.11 (d, J=1.3 Hz, 1H), 8.73 (s, 1H).

Step 20: Preparation of Intermediate trans-10-hydroxy-7-[1-(2-trimethylsilylethoxymethyl)-1H-imidazol-2-yl]-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.SUP.2,6.]undeca-2(6),4-dien-9-one (1t)

(69) To a solution of trans-10-allyloxy-7-[1-(2-trimethylsilylethoxymethyl)-1H-imidazol-2-yl]-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2,6]undeca-2(6),4-dien-9-one (1s) (43 mg, 0.10 mmol) in anhydrous DCM (2 mL) under inert atmosphere were successively added AcOH (0.01 mL, 0.20 mmol) and Pd(PPh.sub.3).sub.4 (57 mg, 0.05 mmol). After stirring for 20 min at rt, the mixture was concentrated in vacuo. The residue was purified by flash chromatography on silica gel (DCM/Acetone 80/20 to 0/100) to give trans-10-hydroxy-7-[1-(2-trimethylsilylethoxymethyl)-1H-imidazol-2-yl]-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2,6]undeca-2(6),4-dien-9-one (1t) (18 mg, 0.05 mmol, 46%).

(70) MS m/z ([M+H]) 394.

(71) .sup.1H NMR (300 MHz, CDCl.sub.3): (ppm) 0.00 (s, 9H), 0.91-1.06 (m, 2H), 1.68 (bs, 1H), 3.34 (dd, J=11.3/3.0 Hz, 1H), 3.64 (dd, J=8.7/8.0 Hz, 2H), 3.74 (d, J=11.2 Hz, 1H), 4.59 (d, J=2.7 Hz, 1H), 5.40 (d, J=10.7 Hz, 1H), 5.68 (d, J=10.7 Hz, 1H), 5.95 (s, 1H), 6.98 (d, J=1.3 Hz, 1H), 7.13 (d, J=1.3 Hz, 1H), 8.74 (s, 1H).

Step 21: Preparation of Sodium {trans-9-oxo-7-[1-(2-trimethylsilylethoxymethyl)-1H-imidazol-2-yl]-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.SUP.2,6.]undeca-2(6),4-dien-10-yl}sulfate (1u)

(72) To a solution of trans-10-hydroxy-7-[1-(2-trimethylsilylethoxymethyl)-1H-imidazol-2-yl]-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2,6]undeca-2(6),4-dien-9-one (1t) (18 mg, 0.05 mmol) in anhydrous pyridine (0.5 mL) under inert atmosphere was added sulfur trioxide pyridine complex (30 mg, 0.19 mmol). After stirring for 16 h, the heterogeneous mixture was concentrated in vacuo. DCM was added to the residue and the solids were filtered off. The crude residue was applied on a Dowex sodium form column (Dowex 50WX8 hydrogen form stored with a 2N aqueous NaOH solution and washed until neutral pH with water). The fractions containing the desired compound were combined, freezed and lyophilized. The salt was triturated with DCM and filtered through PTFE filter. The filtrate was eliminated and the solid was solubilized with water MilliQ. The aqueous layer was freezed and lyophilized to give sodium {trans-9-oxo-7-[1-(2-trimethylsilylethoxymethyl)-1H-imidazol-2-yl]-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2,6]undeca-2(6),4-dien-10-yl}sulfate (1u) (10 mg, 0.02 mmol, 43%) as a white powder.

(73) MS m/z ([M+H]) 474.

(74) .sup.1H NMR (400 MHz, D.sub.2O): (ppm) 0.01 (s, 9H), 0.91-1.08 (m, 2H), 3.59-3.67 (m, 2H), 3.72-3.82 (m, 2H), 5.29 (d, J=1.1 Hz, 1H), 5.59 (d, J=11.0 Hz, 1H), 5.74 (d, J=11.0 Hz, 1H), 6.07 (s, 1H), 6.97 (d, J=1.4 Hz, 1H), 7.41 (d, J=1.4 Hz, 1H), 9.00 (s, 1H).

Step 22: Preparation of Sodium and 2,2,2-trifluoroacetate [trans-7-(1H-imidazol-3-ium-2-yl)-9-oxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.SUP.2,6.]undeca-2(6),4-dien-10-yl]sulfate (Example 1)

(75) Sodium {trans-9-oxo-7-[1-(2-trimethylsilylethoxymethyl)-1H-imidazol-2-yl]-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2,6]undeca-2(6),4-dien-10-yl}sulfate (1u) (10 mg, 0.02 mmol) was dissolved in TFA (0.50 mL, 6.53 mmol) at 0 C. under inert atmosphere. After stirring for 2 h at rt, the mixture was concentrated in vacuo. The residue was triturated several times in Et.sub.2O and the filtrate was filtered through PTFE filter. The solid was solubilized with water MilliQ. The aqueous layer was freezed and lyophilized to give sodium and 2,2,2-trifluoroacetate [trans-7-(1H-imidazol-3-ium-2-yl)-9-oxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2,6]undeca-2(6),4-dien-10-yl]sulfate (Example 1) (6.0 mg, 0.012 mmol, 62%) as a white solid.

(76) MS m/z ([M+H]) 344.

(77) .sup.1H NMR (400 MHz, D.sub.2O): (ppm) 3.42 (dd, J=12.3/0.7 Hz, 1H), 3.83 (dd, J=12.3/2.9 Hz, 1H), 5.33 (d, J=2.9 Hz, 1H), 6.26 (s, 1H), 7.51 (s, 2H), 9.12 (s, 1H).

Example 2: Synthesis of Sodium [trans-7-(1,3,4-oxadiazol-2-yl)-9-oxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.SUP.2,6.]undeca-2(6),4-dien-10-yl]sulfate

(78) ##STR00039##

Step 1: Preparation of Intermediate O5-tert-butyl O4-methyl 7-[allyloxy-(2-nitrophenyl)sulfonyl-amino]-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-4,5-dicarboxylate (2a)

(79) To a solution of methyl 7-[allyloxy-(2-nitrophenyl)sulfonyl-amino]-4,5,6,7-tetrahydrothiazolo[4,5-c]pyridine-4-carboxylate (1g) (1.50 g, 3.30 mmol) in anhydrous THF (17 mL) under inert atmosphere at rt were successively added TEA (0.60 mL, 4.29 mmol) and Boc.sub.2O (973 mg, 4.46 mmol). The mixture was stirred at 40 C. for 5 h then at rt for 10 h. Water and EtOAc were added. The aqueous layer was extracted with EtOAc twice. The organic layers were combined and dried over Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (DCM/acetone 100/0 to 90/10) to give O5-tert-butyl O4-methyl 7-[allyloxy-(2-nitrophenyl)sulfonyl-amino]-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-4,5-dicarboxylate (2a) (1.35 g, 2.43 mmol, 74%) as a white solid.

(80) MS m/z ([M+H]) 555.

Step 2: Preparation of Intermediate tert-butyl 7-[allyloxy-(2-nitrophenyl)sulfonyl-amino]-4-(1,3,4-oxadiazol-2-yl)-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-5-carboxylate (2b)

(81) A mixture of O5-tert-butyl O4-methyl 7-[allyloxy-(2-nitrophenyl)sulfonyl-amino]-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-4,5-dicarboxylate (2a) (1.35 g, 2.43 mmol) and hydrazine hydrate (50-60% in water, 0.5 mL) in MeOH (2.5 mL) under inert atmosphere was stirred at rt for 2 days. The mixture was cooled at 0 C. then water and EtOAc were added. The aqueous layer was extracted with EtOAc twice. The organic layers were combined, dried over Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was dissolved in triethylorthoformate (12 mL, 73.1 mmol). AcOH (150 L) was added and the mixture was heated at 110 C. for 20 h. Further AcOH was added at rt and the mixture was heated at 110 C. for further 30 h. The mixture was concentrated in vacuo. The residue was purified by flash chromatography on silica gel (DCM/acetone 100/0 to 90/10) to give tert-butyl 7-[allyloxy-(2-nitrophenyl)sulfonyl-amino]-4-(1,3,4-oxadiazol-2-yl)-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-5-carboxylate (2b) (851 mg, 1.51 mmol, 62%) as a off-white solid.

(82) MS m/z ([M+H]) 565.

Step 3: Preparation of Intermediate N-allyloxy-2-nitro-N-[4-(1,3,4-oxadiazol-2-yl)-4,5,6,7-tetrahydrothiazolo[4,5-c]pyridin-7-yl]benzenesulfonamide (2c)

(83) To a mixture of tert-butyl 7-[allyloxy-(2-nitrophenyl)sulfonyl-amino]-4-(1,3,4-oxadiazol-2-yl)-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-5-carboxylate (2b) (517 mg, 0.92 mmol) in DCM (47 mL) under inert atmosphere, TFA (5.2 mL) was added at rt. The mixture was stirred for 2 h and poured in an ice-cooled 1M aqueous NaOH solution. The aqueous layer was extracted with DCM twice. The organic layers were combined, dried over Na.sub.2SO.sub.4 and concentrated in vacuo to give N-allyloxy-2-nitro-N-[4-(1,3,4-oxadiazol-2-yl)-4,5,6,7-tetrahydrothiazolo[4,5-c]pyridin-7-yl]benzenesulfonamide (2c) (401 mg, 0.86 mmol, 94%) as a beige solid which was used without further purification.

(84) MS m/z ([M+H]) 465.

Step 4: Preparation of Intermediate N-allyloxy-4-(1,3,4-oxadiazol-2-yl)-4,5,6,7-tetrahydrothiazolo[4,5-c]pyridin-7-amine (2d)

(85) To a solution of N-allyloxy-2-nitro-N-[4-(1,3,4-oxadiazol-2-yl)-4,5,6,7-tetrahydrothiazolo[4,5-c]pyridin-7-yl]benzenesulfonamide (2c) (401 mg, 0.86 mmol) in ACN (5.4 mL) under inert atmosphere at rt were successively added PhSH (0.36 mL, 3.45 mmol) and potassium carbonate (835 mg, 6.04 mmol). The mixture was stirred at rt for 2 h then filtered through a pad of silica gel, rinsed with DCM to eliminate apolar impurity. Then the product was eluted with a DCM/MeOH (90/10) mixture. The filtrate was concentrated in vacuo. The residue was purified by flash chromatography on silica gel (DCM/acetone 100/0 to 60/40) to give N-allyloxy-4-(1,3,4-oxadiazol-2-yl)-4,5,6,7-tetrahydrothiazolo[4,5-c]pyridin-7-amine (2d) (165 mg, 0.59 mmol, 68%).

(86) MS m/z ([M+H]) 280.

Step 5: Preparation of Intermediate trans-10-allyloxy-7-(1,3,4-oxadiazol-2-yl)-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.SUP.2,6.]undeca-2(6),4-dien-9-one (2e)

(87) To a solution of N-allyloxy-4-(1,3,4-oxadiazol-2-yl)-4,5,6,7-tetrahydrothiazolo[4,5-c]pyridin-7-amine (2d) (165 mg, 0.59 mmol) in ACN (60 mL) under inert atmosphere at 10 C. was added TEA (0.37 mL, 2.66 mmol). A solution of diphosgene (39 L, 0.33 mmol) in ACN (1.1 mL) was dropwisely added. After stirring for 2 h at 10 C., the mixture was stirred at rt for 20 h. The solvent was evaporated. The residue was diluted in DCM, washed with a 2M aqueous solution of NaH.sub.2PO.sub.4. The organic layers were combined, dried over Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (DCM/acetone 100/0 to 80/20) to give trans-10-allyloxy-7-(1,3,4-oxadiazol-2-yl)-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2,6]undeca-2(6),4-dien-9-one (2e) (64 mg, 0.21 mmol, 36%) as a white solid.

(88) MS m/z ([M+H]) 306.

(89) .sup.1H NMR (300 MHz, CDCl.sub.3): (ppm) 3.60 (dd, J=11.7/2.8 Hz, 1H), 3.76 (dd, J=11.7/0.7 Hz, 1H), 4.40-4.54 (m, 2H), 4.75 (dd, J=2.8/0.7 Hz, 1H), 5.32-5.41 (m, 2H), 5.94-6.09 (m, 2H), 8.52 (s, 1H), 8.75 (s, 1H).

Step 6: Preparation of Sodium [trans-7-(1,3,4-oxadiazol-2-yl)-9-oxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.SUP.2,6.]undeca-2(6),4-dien-10-yl]sulfate (Example 2)

(90) To a solution of trans-10-allyloxy-7-(1,3,4-oxadiazol-2-yl)-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2,6]undeca-2(6),4-dien-9-one (2e) (58 mg, 0.19 mmol) in anhydrous DCM (4.2 mL) under inert atmosphere were successively added AcOH (21 L, 0.37 mmol) and Pd(PPh.sub.3).sub.4 (108 mg, 0.09 mmol). After stirring at rt for 1 h, the mixture was diluted with pyridine (1.1 mL) and sulfur trioxide-pyridine complex (120 mg, 0.76 mmol) was added. The mixture was stirred at rt for 5 h in the dark and then concentrated in vacuo. DCM was added to the residue and the solids were filtered off. The filtrate was concentrated in vacuo and the residue was applied on a Dowex sodium form column (Dowex 50WX8 hydrogen form stored with a 2N aqueous NaOH solution and washed until neutral pH with water). The fractions containing the desired compound were combined, freezed and lyophilized. The residue was purified by flash chromatography on C18 silica gel (water/ACN 98/2) to give sodium [trans-7-(1,3,4-oxadiazol-2-yl)-9-oxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2,6]undeca-2(6),4-dien-10-yl]sulfate (Example 2) (4.0 mg, 0.006 mmol, 3%) as an off-white powder.

(91) MS m/z ([M+H]) 346.

(92) MS m/z ([M+H]) 344.

(93) .sup.1H NMR (400 MHz, D.sub.2O): (ppm) 3.10 (dd, J=14.5/2.5 Hz, 1H), 3.35 (dd, J=14.5/4.0 Hz, 1H), 5.30 (d, J=2.5 Hz, 1H), 5.70 (s, 1H), 8.92 (s, 1H), 8.98 (s, 1H).

Example 3: Synthesis of Sodium and 2,2,2-trifluoroacetate [trans-7-(4H-1,2,4-triazol-4-ium-3-yl)-9-oxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.SUP.2,6.]undeca-2(6),4-dien-10-yl]sulfate

(94) ##STR00040## ##STR00041##

Step 1: Preparation of Intermediate O5-benzyl 7-[allyloxy-(2-nitrophenyl)sulfonyl-amino]-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-4,5-dicarboxylate (3a)

(95) To a solution of O5-benzyl O4-methyl 7-[allyloxy-(2-nitrophenyl)sulfonyl-amino]-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-4,5-dicarboxylate (1h) (2.03 g, 3.44 mmol) in a mixture of THF (21 mL) and water (14 mL) at rt was dropwise added a solution of lithium hydroxide 1N (4.1 mL, 4.13 mmol). After stirring for 30 min, the mixture was diluted with DCM and acidified to pH 1 with a solution of HCl 1N at rt. The product was extracted with DCM and concentrated in vacuo to give O5-benzyl 7-[allyloxy-(2-nitrophenyl)sulfonyl-amino]-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-4,5-dicarboxylate (3a) (1.98 g, 3.44 mmol, quantitative yield) as a white foam which was used without further purification.

(96) MS m/z ([M+H]) 575.

Step 2: Preparation of Intermediate O5-benzyl 7-[allyloxy-(2-nitrophenyl)sulfonyl-amino]-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-4-carboxamide-5-carboxylate (3b)

(97) To a solution of O5-benzyl 7-[allyloxy-(2-nitrophenyl)sulfonyl-amino]-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-4,5-dicarboxylate (3a) (1.98 g, 3.44 mmol) in anhydrous DCM (17 mL) under inert atmosphere at 0 C. were sequentially added TEA (0.58 mL, 4.13 mmol) and trimethylacetyl chloride (0.45 mL, 3.62 mmol) and the mixture was stirred at 0 C. for 45 min. Then the mixture was cooled down to 20 C. and ammonium hydroxide solution 28-30% (0.57 mL, 13.78 mmol) was dropewisely added. The reacting mixture was stirred at 20 C. for 30 min before being diluted with DCM (5.0 mL) and water (2.0 mL), warmed up to rt, extracted with DCM and concentrated in vacuo to give O5-benzyl 7-[allyloxy-(2-nitrophenyl)sulfonyl-amino]-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-4-carboxamide-5-carboxylate (3b) (1.98 g, 3.44 mmol, quantitative yield) as a brown foam which was used without further purification.

(98) MS m/z ([M+H]) 574.

Step 3: Preparation of Intermediate O5-benzyl 7-[allyloxy-(2-nitrophenyl)sulfonyl-amino]-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-4-(NH-1,2,4-triazol-3-yl)-5-carboxylate (3c)

(99) A solution of O5-benzyl 7-[allyloxy-(2-nitrophenyl)sulfonyl-amino]-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-4-carboxamide-5-carboxylate (3b) (1.98 g, 3.44 mmol) in N,N-dimethylformamide dimethyl acetal (9.2 mL, 68.90 mmol) was stirred at 90 C. for 30 min then cooled down to rt and concentrated under reduced pressure. The residue was dissolved in AcOH (17 mL) and hydrazine hydrate (1.07 mL, 34.45 mmol) was carefully added. The mixture was stirred at 80 C. for 20 min then cooled down to rt and concentrated under reduced pressure. The residue was purified by flash chromatography on silica gel (DCM/acetone 50/50) to give O5-benzyl 7-[allyloxy-(2-nitrophenyl)sulfonyl-amino]-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-4-(NH-1,2,4-triazol-3-yl)-5-carboxylate (3c) (3.31 g contaminated by salts) as a brown oil.

(100) MS m/z ([M+H]) 598.

(101) MS m/z ([MH]) 596.

Step 4: Preparation of Intermediate O5-benzyl 7-[allyloxy-(2-nitrophenyl)sulfonyl-amino]-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-4-(N-(2-trimethylsilylethoxymethyl)-1,2,4-triazol-3-yl)-5-carboxylate (3d)

(102) To a solution of O5-benzyl 7-[allyloxy-(2-nitrophenyl)sulfonyl-amino]-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-4-(NH-1,2,4-triazol-3-yl)-5-carboxylate (3c) (3.31 g, 3.44 mmol in theory) in anhydrous DMF (22 mL) under inert atmosphere were successively added (2-(chloromethoxy)ethyl)trimethylsilane (14.7 mL, 83.08 mmol) and TEA (15.4 mL, 110.78 mmol). After stirring at rt for 30 min, the mixture was concentrated in vacuo. The residue was dissolved in DCM, washed with brine, concentrated in vacuo and then purified by flash chromatography on silica gel (DCM/Acetone 100/0 to 70/30) to give O5-benzyl 7-[allyloxy-(2-nitrophenyl)sulfonyl-amino]-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-4-[N-(2-trimethylsilylethoxymethyl)-1,2,4-triazol-3-yl]-5-carboxylate (3d) (2.70 g contaminated by salts) as a brown oil.

(103) MS m/z ([M+H]) 728.

(104) MS m/z ([MH]) 726.

Step 5: Preparation of Intermediate O5-benzyl 7-(allyloxyamino)-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-4-[N-(2-trimethylsilylethoxymethyl)-1,2,4-triazol-3-yl]-5-carboxylate

(105) To a solution of O5-benzyl 7-[allyloxy-(2-nitrophenyl)sulfonyl-amino]-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-4-[N-(2-trimethylsilylethoxymethyl)-1,2,4-triazol-3-yl]-5-carboxylate (3d) (2.70 g, 3.44 mmol in theory) in ACN (17 mL) under inert atmosphere at rt were successively added potassium carbonate (2.38 g, 17.22 mmol) and PhSH (1.77 mL, 17.22 mmol). The mixture was stirred at rt for 2 h then filtered through a fritt, rinsed with DCM and the filtrate was concentrated under reduced pressure. The residue was purified by flash chromatography on silica gel (DCM/acetone 100/0 to 70/30) to give O5-benzyl 7-(allyloxyamino)-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-4-[N-(2-trimethylsilylethoxymethyl)-1,2,4-triazol-3-yl]-5-carboxylate (3e) (1.39 g, 2.57 mmol, 75% over 6 steps) as a yellow oil.

(106) MS m/z ([M+H]) 543.

Step 6: Preparation of Intermediate O5-benzyl 7-[allyloxy(chlorocarbonyl)amino]-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-4-[N-(2-trimethylsilylethoxymethyl)-1,2,4-triazol-3-yl]-5-carboxylate (3f)

(107) To a solution of O5-benzyl 7-(allyloxyamino)-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-4-[N-(2-trimethylsilylethoxymethyl)-1,2,4-triazol-3-yl]-5-carboxylate (3e) (1.39 g, 2.57 mmol) in DCM (26 mL) under inert atmosphere at 0 C. were dropwisely added TEA (0.72 mL, 5.13 mmol) and diphosgene (0.40 mL, 3.38 mmol). After stirring at 0 C. for 30 min, the mixture was diluted with DCM, washed with brine and concentrated under reduced pressure to give O5-benzyl 7-[allyloxy(chlorocarbonyl)amino]-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-4-[N-(2-trimethylsilylethoxymethyl)-1,2,4-triazol-3-yl]-5-carboxylate (3f) (1.55 g, 2.57 mmol, quantitative yield) as a brown oil.

(108) MS m/z ([M+H]) 605 (.sup.39Cl)/607 (.sup.41Cl).

Step 7: Preparation of Intermediate trans-10-allyloxy-7-(NH-1,2,4-triazol-3-yl)-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.SUP.2,6.]undeca-2(6),4-dien-9-one (3g)

(109) To a solution of O5-benzyl 7-[allyloxy(chlorocarbonyl)amino]-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-4-[N-(2-trimethylsilylethoxymethyl)-1,2,4-triazol-3-yl]-5-carboxylate (3f) (1.55 g, 2.57 mmol) in anhydrous DCM (13 mL) under inert atmosphere at rt was dropwisely added TFA (12.8 mL, 167.68 mmol). After stirring at rt for 2 h, the reaction mixture was cooled down to 0 C. and a solution of methanesufonic acid (3.33 mL, 51.34 mmol) in DCM (26 mL) was dropewisely added. The mixture was stirred at 0 C. for 24 h, then diluted with DCM (20 mL), carefully added to a solution of TEA (18 mL) in DCM (100 mL) at 0 C., warmed up to rt and stirred for additional 20 min. The mixture was filtered through a pad of Celite and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (DCM/acetone 50/50 to 0/100) to give trans-10-allyloxy-7-(NH-1,2,4-triazol-3-yl)-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2,6]undeca-2(6),4-dien-9-one (3g) (781 mg polluted by a non-UV visible impurity) as a yellow oil.

(110) MS m/z ([M+H]) 305.

(111) MS m/z ([MH]) 303.

Step 8: Preparation of Intermediate trans-10-allyloxy-7-[N-(2-trimethylsilylethoxymethyl)-1,2,4-triazol-3-yl]-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.SUP.2,6.]undeca-2(6),4-dien-9-one (3h)

(112) To a solution of trans-10-allyloxy-7-(NH-1,2,4-triazol-3-yl)-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2,6]undeca-2(6),4-dien-9-one (3g) (781 mg, 2.57 mmol in theory) in anhydrous DMF (5.1 mL) under inert atmosphere at 0 C. were successively added (2-(chloromethoxy)ethyl)trimethylsilane (1.1 mL, 6.42 mmol) and TEA (1.8 mL, 12.83 mmol). After stirring at rt for 17 h, the mixture was concentrated in vacuo. The residue was dissolved in DCM, washed with brine, concentrated in vacuo and then purified by flash chromatography on silica gel (DCM/Acetone 100/0 to 50/50) to give trans-10-allyloxy-7-[N-(2-trimethylsilylethoxymethyl)-1,2,4-triazol-3-yl]-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2,6]undeca-2(6),4-dien-9-one (3h) (117 mg, 0.27 mmol, 10% over 2 steps) as a brown oil.

(113) MS m/z ([M+H]) 435.

Step 9: Preparation of Intermediate trans-10-hydroxy-7-[N-(2-trimethylsilylethoxymethyl)-1,2,4-triazol-3-yl]-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.SUP.2,6.]undeca-2(6),4-dien-9-one (3i)

(114) To a solution of trans-10-allyloxy-7-[N-(2-trimethylsilylethoxymethyl)-1,2,4-triazol-3-yl]-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2,6]undeca-2(6),4-dien-9-one (3h) (117 mg, 0.27 mmol) in anhydrous DCM (2.7 mL) under inert atmosphere at rt were successively added AcOH (0.03 mL, 0.54 mmol) and Pd(PPh.sub.3).sub.4 (155 mg, 0.14 mmol). After stirring for at rt 30 min, the mixture was concentrated in vacuo. The residue was purified by flash chromatography on silica gel (DCM/Acetone 80/20 to 0/100) to give trans-10-hydroxy-7-[N-(2-trimethylsilylethoxymethyl)-1,2,4-triazol-3-yl]-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2,6]undeca-2(6),4-dien-9-one (3i) (43 mg, 0.11 mmol, 41%) as a yellow foam.

(115) MS m/z ([M+H]) 395.

Step 10: Preparation of Intermediate Sodium {trans-9-oxo-7-[N-(2-trimethylsilylethoxymethyl)-1,2,4-triazol-3-yl]-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.SUP.2,6.]undeca-2(6),4-dien-10-yl}sulfate (3j)

(116) To a solution of trans-10-hydroxy-7-[N-(2-trimethylsilylethoxymethyl)-1,2,4-triazol-3-yl]-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2,6]undeca-2(6),4-dien-9-one (3i) (43 mg, 0.11 mmol) in anhydrous pyridine (1.1 mL) under inert atmosphere at rt was added sulfur trioxide pyridine complex (87 mg, 0.55 mmol). After stirring at rt for 17 h, the heterogeneous mixture was concentrated in vacuo. DCM was added to the residue, the solids were filtered off and the filtrate was concentrated in vacuo. The crude residue was applied on a Dowex sodium form column (Dowex 50WX8 hydrogen form stored with an aqueous solution of 2N NaOH and washed until neutral pH with water). The fractions containing the desired compound were combined, freezed and lyophilized. The salt was triturated with DCM and filtered through PTFE filter. The filtrate was eliminated and the solid was solubilized with water MilliQ. The aqueous layer was freezed and lyophilized to give sodium {trans-9-oxo-7-[N-(2-trimethylsilylethoxymethyl)-1,2,4-triazol-3-yl]-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2,6]undeca-2(6),4-dien-10-yl}sulfate (3j) (28 mg, 0.06 mmol, 52%) as a light-brown powder.

(117) MS m/z ([M+H]) 475.

(118) MS m/z ([MH]) 473.

Step 11: Preparation of Intermediate Sodium and 2,2,2-trifluoroacetate [trans-7-(NH-1,2,4-triazol-4-ium-3-yl)-9-oxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.SUP.2,6.]undeca-2(6),4-dien-10-yl]sulfate (Example 3)

(119) Sodium {trans-9-oxo-7-[N-(2-trimethylsilylethoxymethyl)-1,2,4-triazol-3-yl]-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2,6]undeca-2(6),4-dien-10-yl}sulfate (3j) (28 mg, 0.06 mmol) was dissolved in anhydrous DCM (0.28 mL) and TFA (0.28 mL, 3.65 mmol) was dropewisely added at 0 C. under inert atmosphere. After stirring for 1 h at rt, the mixture was diluted in DCM (1.0 mL) and concentrated in vacuo. The residue was triturated several times in Et.sub.2O then in ACN and the filtrate was filtered through PTFE filter. The solid was solubilized with water MilliQ. The aqueous layer was freezed and lyophilized to give sodium and 2,2,2-trifluoroacetate [trans-7-(NH-1,2,4-triazol-4-ium-3-yl)-9-oxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2,6]undeca-2(6),4-dien-10-yl]sulfate (Example 3) (10 mg, 0.021 mmol, 35%) as a beige solid.

(120) MS m/z ([M+H]) 345.

(121) MS m/z ([MH]) 343.

(122) .sup.1H NMR (300 MHz, D.sub.2O): (ppm) 3.62-3.70 (m, 2H), 5.27-5.32 (m, 1H), 5.97 (s, 1H), 8.51 (s, 1H), 9.00 (s, 1H).

(123) .sup.19F NMR (282 MHz, D.sub.2O): (ppm) 75.57 (s, 3F).

Example 4: Synthesis of Sodium [trans-7-(1,2,4-oxadiazol-5-yl)-9-oxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.SUP.2,6.]undeca-2(6),4-dien-10-yl]sulfate

(124) ##STR00042##

Step 1: Preparation of Intermediate O5-benzyl 7-[allyloxy-(2-nitrophenyl)sulfonyl-amino]-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-4-(1,2,4-oxadiazol-5-yl)-5-carboxylate (4a)

(125) A solution of O5-benzyl 7-[allyloxy-(2-nitrophenyl)sulfonyl-amino]-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-4-carboxamide-5-carboxylate (3b) (2.60 g, 4.53 mmol) in N,N-dimethylformamide dimethyl acetal (12.0 mL, 90.66 mmol) was stirred at 90 C. for 30 min then cooled down to rt and concentrated under reduced pressure. The residue was dissolved in AcOH (23 mL) and a commercial hydroxylamine solution 50 wt. % in H.sub.2O (2.78 mL, 45.33 mmol) was carefully added. The mixture was stirred at 90 C. for 30 min then cooled down to rt and concentrated under reduced pressure. The residue was partitioned between DCM and water, extracted with DCM and concentrated under reduced pressure. The residue was purified by flash chromatography on silica gel (DCM/acetone 80/20) to give O5-benzyl 7-[allyloxy-(2-nitrophenyl)sulfonyl-amino]-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-4-(1,2,4-oxadiazol-5-yl)-5-carboxylate (4a) (1.66 g, 2.78 mmol, 61% over two steps) as a light-yellow foam.

(126) MS m/z ([M+H]) 599.

(127) MS m/z ([MH]) 597.

Step 2: Preparation of Intermediate O5-benzyl 7-[allyloxyamino]-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-4-(1,2,4-oxadiazol-5-yl)-5-carboxylate (4b)

(128) To a solution of O5-benzyl 7-[allyloxy-(2-nitrophenyl)sulfonyl-amino]-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-4-(1,2,4-oxadiazol-5-yl)-5-carboxylate (4a) (1.66 g, 2.78 mmol) in ACN (28 mL) under inert atmosphere at rt were successively added potassium carbonate (1.92 g, 13.88 mmol) and PhSH (0.86 mL, 8.33 mmol). The mixture was stirred at rt for 1 h then filtered through a fritt, rinsed with DCM and the filtrate was concentrated under reduced pressure. The residue was purified by flash chromatography on silica gel (DCM/acetone 90/10) to give O5-benzyl 7-(allyloxyamino)-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-4-(1,2,4-oxadiazol-5-yl)-5-carboxylate (4b) (805 mg, 1.95 mmol, 70%) as a light-yellow oil.

(129) MS m/z ([M+H]) 414.

(130) MS m/z ([MH]) 412.

Step 3: Preparation of Intermediate O5-benzyl 7-[allyloxy(chlorocarbonyl)amino]-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-4-(1,2,4-oxadiazol-5-yl)-5-carboxylate (4c)

(131) To a solution of O5-benzyl 7-(allyloxyamino)-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-4-(1,2,4-oxadiazol-5-yl)-5-carboxylate (4b) (805 mg, 1.95 mmol) in DCM (20 mL) under inert atmosphere at 0 C. were dropwisely added TEA (0.54 mL, 3.89 mmol) and diphosgene (0.31 mL, 2.53 mmol). After stirring at 0 C. for 30 min, the mixture was diluted with DCM, washed with brine and concentrated under reduced pressure to give O5-benzyl 7-[allyloxy(chlorocarbonyl)amino]-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-4-(1,2,4-oxadiazol-5-yl)-5-carboxylate (4c) (930 mg, 1.95 mmol, quantitative yield) as a brown oil.

(132) MS m/z ([M+H]) 476.

Step 4: Preparation of Intermediate trans-10-allyloxy-7-(1,2,4-oxadiazol-5-yl)-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.SUP.2,6.]undeca-2(6),4-dien-9-one (4d)

(133) To a solution of O5-benzyl 7-[allyloxy(chlorocarbonyl)amino]-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-4-(1,2,4-oxadiazol-5-yl)-5-carboxylate (4c) (930 mg, 1.95 mmol) in anhydrous DCM (20 mL) under inert atmosphere at 0 C. was dropwisely added methanesulfonic acid (2.53 mL, 38.94 mmol). After stirring at 0 C. for 24 h, the reaction mixture was diluted with DCM, carefully added to a solution of pyridine (8.0 mL) in DCM (8.0 mL) at 0 C., warmed up to rt and stirred for additional 10 min. The mixture was filtered through a pad of Celite and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (DCM/acetone 90/10) to give trans-0-allyloxy-7-(1,2,4-oxadiazol-5-yl)-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2,6]undeca-2(6),4-dien-9-one (4d) (318 mg, 1.04 mmol, 54%) as a yellow oil.

(134) MS m/z ([M+H]) 306.

(135) MS m/z ([M+H]) 304.

(136) .sup.1H NMR (400 MHz, CDCl.sub.3): (ppm) 3.65 (dd, J=12.0/4.0 Hz, 1H), 3.74 (d, J=12.0 Hz, 1H), 4.42-4.53 (m, 2H), 4.75 (d, J=2.8 Hz, 1H), 5.33-5.36 (m, 1H), 5.36-5.42 (m, 1H), 5.97-6.07 (m, 1H), 6.11 (s, 1H), 8.44 (s, 1H), 8.75 (s, 1H).

Step 5: Preparation of Sodium [trans-7-(1,2,4-oxadiazol-5-yl)-9-oxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.SUP.2,6.]undeca-2(6),4-dien-10-yl]sulfate (Example 4)

(137) To a solution of trans-10-allyloxy-7-(1,2,4-oxadiazol-5-yl)-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2,6]undeca-2(6),4-dien-9-one (4d) (111 mg, 0.36 mmol) in anhydrous DCM (3.6 mL) under inert atmosphere were successively added AcOH (0.04 mL, 0.73 mmol) and Pd(PPh.sub.3).sub.4 (210 mg, 0.18 mmol). After stirring at rt for 1 h, the mixture was diluted with anhydrous pyridine (3.6 mL) and sulfur trioxide pyridine complex (289 mg, 1.82 mmol) was added in one portion. After stirring at rt for 48 h, the heterogeneous mixture was concentrated in vacuo. The residue was diluted with DCM, the solids were filtered off and the filtrate was concentrated in vacuo. The residue was purified by flash chromatography on silica gel (DCM/acetone 70/30 to 100% acetone) and, after concentration in vacuo, the obtained light-yellow foam residue was applied on a Dowex sodium form column (Dowex 50WX8 hydrogen form stored with an aqueous solution of 2N NaOH and washed until neutral pH with water). The fractions containing the desired compound were combined, freezed and lyophilized. The salt was triturated with DCM and filtered through PTFE filter. The filtrate was eliminated and the solid was solubilized with water MilliQ. The aqueous layer was freezed and lyophilized to give sodium [trans-7-(1,2,4-oxadiazol-5-yl)-9-oxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2,6]undeca-2(6),4-dien-10-yl]sulfate (Example 4) (14.5 mg, 0.04 mmol, 11%) as a white powder.

(138) MS m/z ([M+H]) 346.

(139) MS m/z ([M+H]) 344.

(140) .sup.1H NMR (400 MHz, D.sub.2O): (ppm) 3.67 (d, J=12.0 Hz, 1H), 3.86 (dd, J=12.0/4.0 Hz, 1H), 5.33 (d, J=3.0 Hz, 1H), 6.03 (s, 1H), 8.79 (s, 1H), 9.06 (s, 1H).

Example 5: Biological Activity

(141) Method 1: -Lactamase Inhibitory Activity, Determination of IC.sub.50 (Table 1)

(142) Enzyme activity was monitored by spectrophotometric measurement of nitrocefin (NCF-TOKU-E, N005) hydrolysis at 485 nm, at room temperature and in assay buffer A: 100 mM Phosphate pH7, 2% glycerol and 0.1 mg/mL Bovine serum albumin (Sigma, B4287). Enzymes were cloned in E. coli expression vector, expressed and purified in house using classical procedures. To a transparent polystyrene plate (Corning, 3628) were added in each well 5 L DMSO or inhibitor dilutions in DMSO and 80 L enzyme in buffer A. Plates were immediately read at 485 nm in a microplate spectrophotometer (BioTek, PowerWave HT) to enable background subtraction. After 30 min of pre-incubation at room temperature, 15 L of NCF (200 M final) were finally added in each well. Final enzyme concentrations were 0.1 nM (TEM-1), 0.075 nM (SHV-1), 0.4 nM (CTX-M-15), 1 nM (KPC-2), 0.2 nM (P99 AmpC), 0.2 nM (CMY-37), 0.4 nM (AmpC P. aeruginosa), 0.2 nM (OXA-1), 1.2 nM (OXA-11), 0.4 nM (OXA-15) and 0.3 nM (OXA-48). After 20 min incubation at room temperature, plates were once again read at 485 nm. Enzyme activity was obtained by subtracting the final signal by the background, and was converted to enzyme inhibition using non inhibited wells. IC.sub.50 curves were fitted to a classical Langmuir equilibrium model with Hill slope using XLFIT (IDBS).

(143) TABLE-US-00001 TABLE 1 IC.sub.50 (nM) for -lactamase Inhibitory Activity IC.sub.50 -lactamase (M) (A) (C) CTX-M- AmpC AmpC (D) TEM-1 SHV-1 15 KPC-2 (P99) CMY-37 (PAE) OXA-1 OXA-11 OXA-15 OXA-48 Example 1 0.012 0.0055 0.0027 0.0042 0.00037 0.00063 0.017 0.19 0.41 0.27 0.0084 Example 3 0.046 0.14 0.028 0.46 0.013 0.030 0.57 1.0 0.19 1.1 0.0068 Example 4 0.0011 0.0014 0.00064 0.0032 0.0011 0.0034 0.064 0.17 0.0065 0.019 0.00034

(144) Method 2: MIC of Compounds and Synergy with Ceftazidime Against Bacterial Isolates (Table 2 and 3)

(145) Compounds of the present invention were assessed against genotyped bacterial strains alone or in combination with the -lactam ceftazidime. In the assays, MICs of said compounds, or of ceftazidime at fixed concentrations of said compounds were determined by the broth microdilution method according to the Clinical Laboratory Standards Institute (CLSIM7-A7). Briefly, compounds alone according to the invention were prepared in DMSO and spotted (2 L each) on sterile polystyrene plates (Corning, 3788). Compounds and ceftazidime dilutions were prepared in DMSO and spotted (1 L each) on sterile polystyrene plates (Corning, 3788). Log phase bacterial suspensions were adjusted to a final density of 510.sup.5 cfu/mL in cation-adjusted Mueller-Hinton broth (Beckton-Dickinson) and added to each well (98 L). Microplates were incubated for 16-20 h at 35 C. in ambient air. The MIC of of the compounds was defined as the lowest concentration of said compounds that prevented bacterial growth as read by visual inspection. The MIC of ceftazidime at each compound concentration was defined as the lowest concentration of ceftazidime that prevented bacterial growth as read by visual inspection.

(146) TABLE-US-00002 TABLE 2 Bacterial species used in MIC determination CFR C. freundii CMU C. murliniae EAE E. aerogenes ECL E. cloacae ECO E. coli KOX K. oxytoca KPN K. pneumoniae MMO M. morganii PAE P. aeruginosa PMI P. mirabilis SMA S. marcescens

(147) TABLE-US-00003 TABLE 3 MIC of compounds and Ceftazidime/compound combinations MIC (g/ml) CAZ + CAZ + CAZ + beta- Example 1 Example 3 Example 4 Strain lactamases CAZ Example 1 (4 g/mL) Example 3 (4 g/mL) Example 4 (4 g/mL) ECO 190317 TEM-1, SHV-12, 128 1 <0.25 16 32 16 0.25 CTX-M-15 OXA-1 ECO 180070 TEM-1, CTX-M-15 64 1 <0.25 ECO 190314 CTX-M-1 8 0.5 <0.25 ECO 190457 CTX-M-15 OXA-1 16 1 <0.25 ECO 190549 CTX-M-1 8 0.25 <0.25 ECO 200159 TEM-1, CTX-M-14 2 0.25 <0.25 ECO 200259 CTX-M-14 4 0.5 <0.25 ECO 200344 CTX-M-1 16 0.5 <0.25 ECO 260096 CTX-M-132 128 4 <0.25 ECO 260304 CTX-M-15 16 1 <0.25 ECO 260508 TEM-1, CTX-M-15 128 2 <0.25 OXA-1 ECO UFR16 TEM-1, CTX-M-15 64 2 <0.25 OXA-1 OXA-48 ECO UFR17 TEM-1, CTX-M-15 >128 4 <0.25 CMY-2 OXA-1 OXA-181 ECO UFR18 CTX-M-15 OXA- >128 1 <0.25 204 ECO UFR19 CTX-M-15 CMY-2 128 1 <0.25 OXA-1 OXA-204 ECO UFR32 TEM-1, VEB-1 >128 1 <0.25 OXA-10 ECO UFR39 CTX-M-15 >128 2 <0.25 NDM-1 ECO UFR41 TEM-1, CTX-M-15 >128 16 >128 CMY-2 OXA-1 NDM-4 ECO UFR45 TEM-1 CMY-4 2 0.5 <0.25 OXA-1 OXA-48 VIM-1 ECO UFR52 TEM-1, SHV-12 >128 1 <0.25 IMP-8 ECO UFR61O TEM-1 KPC-2 128 1 <0.25 ECO UFR62 TEM-1, CTX-M-9 16 0.5 <0.25 KPC-2 ECO UFR74 SHV-1 DHA-1 128 2 <0.25 ECO UFR86 2 2 <0.25 ECL P99 AmpC 128 4 <0.25 32 2 EAE 200261 TEM-x AmpC 128 8 <0.25 ECL NEM146383 AmpC >128 2 <0.25 ECL 190310 AmpC >128 4 <0.25 ECL 190408 TEM-1, CTX-M-15 128 0.5 <0.25 OXA-1 ECL 200138 AmpC >128 8 8 ECL 200322 TEM-1, CTX-M-15 >128 1 <0.25 OXA-1 ECL 260033 AmpC >128 8 4 ECL 260253 TEM-1 KPC-3 >128 8 0.5 ECL 260323 AmpC >128 4 <0.25 ECL 260508 TEM-1, CTX-M-15 64 1 <0.25 ECL HAN OXA-35 >128 8 2 ECL 2185D OXA-163 128 8 1 ECL UFR12 CTX-M-9 OXA-48 2 2 <0.25 ECL UFR13 TEM-1, SHV-12, >128 2 <0.25 CTX-M-9 OXA-48 ECL UFR14 TEM-1, SHV-12, >128 8 <0.25 CTX-M-15 DHA-1 OXA-1 OXA-48 ECL UFR38 CTX-M-15 >128 1 <0.25 NDM-1 ECL UFR51 SHV-12 IMP-8 >128 0.5 <0.25 ECL UFR60 TEM-1, CTX-M-15 >128 8 <0.25 KPC-2 ECL UFR70 TEM-1, CTX-M-15 >128 1 <0.25 CMY-2 OXA-1 ECL UFR84 TEM-1 AmpC >128 8 <=0.25 OXA-1 ECL UFR85 TEM-1, CTX-M-15 >128 2 <0.25 AmpC KPN 6299 TEM-1, SHV-11 256 16 <=0.125 >32 2 OXA-163 KPN BAA- TEM-1, SHV-11, 256 4 0.5 1898 SHV-12 KPC-2 KPN 700603 SHV-18 OXA-2 64 4 <0.25 KPN 110376 TEM-1, SHV-1, 128 2 <0.25 CTX-M-15 OXA-1 OXA-48 KPN 131119 TEM-1, SHV-11, >128 4 <=0.25 CTX-M-15 OXA-1 OXA-48 KPN 160143 TEM-1, SHV-1, 128 4 <0.25 CTX-M-15 KPC-2 OXA-1 KPN 190128 TEM-1, SHV-32, >128 4 <0.25 CTX-M-15 OXA-1 KPN 190270 TEM-1, SHV-76, >128 8 <0.25 CTX-M-15 OXA-1 KPN 190425 TEM-1, SHV-1, >128 4 <0.25 CTX-M-15 OXA-1 KPN 190551 TEM-1, SHV-1, 64 2 <0.25 CTX-M-15 OXA-1 KPN 200047 TEM-1, SHV-32, 128 2 <0.25 CTX-M-15 OXA-1 KPN 200327 TEM-1, SHV-1, 32 2 <0.25 CTX-M-15 OXA-1 KPN 260251 TEM-1, SHV-11, >128 4 <0.25 SHV-12, CTX-M-15 KPC-2 KPN 260252 TEM-1, SHV-11 >128 >32 >128 KPC-3 KPN 260376 SHV-1, SHV-49 128 2 <0.25 OXA-1 KPN 270077 TEM-1, SHV-1, 128 8 <0.25 CTX-M-15 KPN 6122012 SHV-1 NDM-1 >128 8 <0.25 KPN ARA TEM-1, SHV-11, >128 4 <0.25 CTX-M-15 OXA-1 OXA-48 KPN LIB SHV-11 OXA-48 0.25 2 <0.25 KOX UFR21 TEM-1, CTX-M-15 128 4 <0.25 OXA-48 KPN UFR22_O TEM-1, SHV-1 2 4 <0.25 OXA-48 KPN UFR24 TEM-1, SHV-2, >128 8 <0.25 SHV-11 OXA-1 OXA-48 OXA-47 KPN UFR25 TEM-1, SHV-11, 128 2 <0.25 CTX-M-15 OXA- 162 KPN UFR27 TEM-1, SHV-28, >128 4 <0.25 CTX-M-15 OXA- 204 KPN UFR28 TEM-1, SHV-1, 128 2 <0.25 CTX-M-15 OXA-1 OXA-232 KPN UFR42 SHV-2, CTX-M-15 >128 4 <0.25 OXA-1 OXA-181 NDM-1 KPN UFR43 SHV-11, CTX-M-15 >128 16 <=0.25 CMY-2 OXA-1 NDM-1 KPN UFR53 TEM-1 IMP-1 >128 4 <0.25 KPN UFR65 TEM-1, SHV-11 128 16 2 KPC-2 KPN UFR66 TEM-1, SHV-11, >128 8 <=0.25 CTX-M-15 KPC-2 KPN UFR68 TEM-1, SHV-11, >128 8 <=0.25 CTX-M-15 KPC-3 KPN UFR77 CMY-2 64 4 <0.25 KPN UFR79 DHA-1 OXA-1 32 4 <0.25 KPN UFR80 SHV-11 DHA-1 0.5 1 <0.25 OXA-1 KPN UFR81 TEM-1, SHV-1 128 32 <=0.25 DHA-1 OXA-48 MMO 200321 TEM-1, CTX-M-15 16 32 <=0.25 OXA-1 CMU 210102 VIM-4 >128 4 <0.25 CFR UFR10 OXA-48 64 1 <0.25 CFR UFR11 TEM-1 OXA-1 8 8 1 OXA-48 SMA UFR30 OXA-405 16 16 <0.25 CFR UFR37 TEM-1, CTX-M-15 >128 2 <0.25 NDM-1 PMI UFR82 CMY-2 4 32 <=0.25 PAE CIP107051 TEM-24 256 >32 8 PAE MUS OXA-20 OXA-18 128 >32 8