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) ##STR00045## wherein: W, unsubstituted or substituted by one or more T, is a non-aromatic, unsaturated 5- or 6-member heterocycle comprising at least one NR.sup.2 group and a (X).sub.n group; X, identical or different, is independently selected from the group consisting of represents C(O), O, N, N(R.sup.2), S, S(O), and S(O).sub.2; R.sup.1 is selected from the group consisting of a carbon-linked 4- or 5- or 6-member heterocycle comprising at least one nitrogen atom that is aromatic or saturated or totally unsaturated or partially unsaturated and optionally substituted by one or more T.sup.1, H, CN, C(O)NHQ.sup.1, C(O)NHOQ.sup.1, C(O)NHNHQ.sup.1, C(O)ONHQ.sup.1, C(O)OQ.sup.1, (CH.sub.2).sub.mOC(O)OQ.sup.1, (CH.sub.2).sub.mOQ.sup.1, (CH.sub.2).sub.mOC(O)Q.sup.1, (CH.sub.2).sub.mOC(O)NQ.sup.1Q.sup.2, (CH.sub.2).sub.mNHC(O)Q.sup.1, (CH.sub.2).sub.mNHS(O).sub.2Q.sup.1, (CH.sub.2).sub.mNHS(O).sub.2NQ.sup.1Q.sup.2, (CH.sub.2).sub.mNHC(O)OQ.sup.1; (CH.sub.2).sub.mNHC(O)NQ.sup.1Q.sup.2, (CH.sub.2).sub.mNHQ.sup.3, (CH.sub.2).sub.mNHC(NHQ.sup.3)NQ.sup.4, (CH.sub.2).sub.mNHCHNQ.sup.3, and C(NHQ.sup.3)NQ.sup.4; R.sup.2 is one of the following: R2, identical or different, is independently selected from the group consisting of (CH.sub.2).sub.qOQ.sup.5, C(O)(CH.sub.2).sub.vOQ.sup.5, (C(O)).sub.w(CH.sub.2).sub.vCN, (CH.sub.2).sub.qOC(O)Q.sup.5, C(O)(CH.sub.2).sub.vOC(O)Q.sup.5, (C(O)).sub.w(CH.sub.2).sub.vC(O)OQ.sup.5, (CH.sub.2).sub.qOC(O)OQ.sup.5, C(O)(CH.sub.2).sub.vOC(O)OQ.sup.5, (CH.sub.2).sub.qOC(O)NQ.sup.5Q.sup.6, C(O)(CH.sub.2).sub.vOC(O)NQ.sup.5Q.sup.6, (C(O)).sub.w(CH.sub.2).sub.vC(O)NQ.sup.5Q.sup.6, (C(O)).sub.w(CH.sub.2).sub.vC(O)ONQ.sup.5, (C(O)).sub.w(CH.sub.2).sub.vC(O)NHOQ.sup.5, (C(O)).sub.w(CH.sub.2).sub.vC(O)NHNHQ.sup.5, (C(O)).sub.w(CH.sub.2).sub.vC(O)ONHQ.sup.5, (CH.sub.2).sub.qNHC(O)Q.sup.5, C(O)(CH.sub.2).sub.vNHC(O)Q.sup.5, (CH.sub.2).sub.qNHS(O).sub.2Q.sup.5, C(O)(CH.sub.2).sub.vNHS(O).sub.2Q.sup.5, (CH.sub.2).sub.qNHS(O).sub.2NQ.sup.5Q.sup.6, C(O)(CH.sub.2).sub.vNHS(O).sub.2NQ.sup.5Q.sup.6, (CH.sub.2).sub.qNHC(O)OQ.sup.5, C(O)(CH.sub.2).sub.vNHC(O)OQ.sup.5, (CH.sub.2).sub.qNHC(O)NQ.sup.5Q.sup.6, C(O)(CH.sub.2).sub.vNHC(O)NQ.sup.5Q.sup.6, (CH.sub.2).sub.qNQ.sup.5Q.sup.6, C(O)(CH.sub.2).sub.vNQ.sup.5Q.sup.6, (CH.sub.2).sub.qNHC(NHQ.sup.3)NQ.sup.4, C(O)(CH.sub.2).sub.vNHC(NHQ.sup.3)NQ.sup.4, (CH.sub.2).sub.qNHCHNQ.sup.3, C(O)(CH.sub.2).sub.vNHCHNQ.sup.3, (C(O)).sub.w(CH.sub.2).sub.vC(NHQ.sup.3)NQ.sup.4, C(O)NQ.sup.5Q.sup.6, and C(NHQ.sup.3)NQ.sup.4; or R.sup.2, identical or different, is independently selected from the group consisting of (C(O)).sub.wC.sub.1-C.sub.3-alkyl that is optionally substituted by one or more T.sup.2, (C(O)).sub.wC.sub.1-C.sub.3-fluoroalkyl that is optionally substituted by one or more T.sup.2, (C(O)).sub.w(CH.sub.2).sub.pC.sub.3-C.sub.6-cycloalkyl that is optionally substituted by one or more T.sup.2, (C(O)).sub.w(CH.sub.2).sub.pC.sub.3-C.sub.6-cyclofluoroalkyl that is optionally substituted by one or more T.sup.2, and (C(O)).sub.w(CH.sub.2).sub.p-(4- or 5- or 6-member aromatic or saturated or totally unsaturated or partially unsaturated heterocycle) that is optionally substituted by one or more T.sup.2; R.sup.3 is selected from the group consisting of SO.sub.3H, CFHCO.sub.2H, and CF.sub.2CO.sub.2H; Q.sup.1 and Q.sup.2 are one of the following: Q.sup.1 and Q.sup.2, identical or different, are independently selected from the group consisting of H, (CH.sub.2).sub.qNHQ.sup.3, (CH.sub.2).sub.qNHC(NHQ.sup.3)NQ.sup.4, (CH.sub.2).sub.qNHCHNQ.sup.3, (CH.sub.2).sub.vC(NHQ.sup.3)NQ.sup.4, (CH.sub.2).sub.qOQ.sup.3, and (CH.sub.2).sub.vCONHQ.sup.3; or Q.sup.1 and Q.sup.2, identical or different, are independently elected from the group consisting of a C.sub.1-C.sub.3 alkyl optionally substituted by one or more T.sup.2 and (CH.sub.2).sub.p-(4- or 5- or 6-member heterocycle) optionally substituted by one or more T.sup.2; or Q.sup.1 and Q.sup.2 and the nitrogen atom to which they are bonded, form a saturated or partially unsaturated 4- or 5- or 6-member heterocycle comprising 1 or 2 or 3 or 4 heteroatoms; Q.sup.3 and Q.sup.4, identical or different, are independently selected from the group consisting of H and a C.sub.1-C.sub.3 alkyl; Q.sup.5 and Q.sup.6 are one of the following: Q.sup.5 and Q.sup.6, identical or different, are independently selected from the group consisting of H, (CH.sub.2).sub.qNHQ.sup.3, (CH.sub.2).sub.qNHC(NHQ.sup.3)NQ.sup.4, (CH.sub.2).sub.qNHCHNQ.sup.3, (CH.sub.2).sub.v C(NHQ.sup.3)NQ.sup.4, (CH.sub.2).sub.qOQ.sup.3; and (CH.sub.2).sub.vCONHQ.sup.3; or Q.sup.5 and Q.sup.6, identical or different, are independently selected from the group consisting of a C.sub.1-C.sub.4 alkyl optionally substituted by one or more T.sup.2 and (CH.sub.2).sub.p-(4- or 5- or 6-member heterocycle) optionally substituted by one or more T.sup.2; or Q.sup.5 and Q.sup.6 and the nitrogen atom to which they are bonded form a saturated or partially unsaturated 4- or 5- or 6-member heterocycle comprising 1 or 2 or 3 or 4 heteroatoms; T.sup.1 is one of the following: T.sup.1, identical or different, is independently selected from the group consisting of F, (CH.sub.2).sub.pOQ.sup.1, (CH.sub.2).sub.pCN, (CH.sub.2).sub.pOC(O)Q.sup.1, (CH.sub.2).sub.pC(O)OQ.sup.1, (CH.sub.2).sub.pOC(O)OQ.sup.1, (CH.sub.2).sub.pOC(O)NHQ.sup.1, (CH.sub.2).sub.pC(O)NHQ.sup.1, (CH.sub.2).sub.pC(O)NHOQ.sup.1, CH.sub.2).sub.pC(O)NHNHQ.sup.1, (CH.sub.2).sub.pC(O)ONHQ.sup.1, (CH.sub.2).sub.pNHC(O)Q.sup.1, (CH.sub.2).sub.pNHS(O).sub.2Q.sup.1, (CH.sub.2).sub.pNHS(O).sub.2NQ.sup.1Q.sup.2, (CH.sub.2).sub.pNHC(O)OQ.sup.1, (CH.sub.2).sub.pNHC(O)NQ.sup.1Q.sup.2, (CH.sub.2).sub.pNHQ.sup.1, (CH.sub.2).sub.pNHC(NHQ.sup.3)NQ.sup.4, (CH.sub.2).sub.pNHCHNQ.sup.3, and (CH.sub.2).sub.pC(NHQ.sup.3)NQ.sup.4; or T.sup.1, identical or different, is independently selected from the group consisting of C.sub.1-C.sub.3 alkyl optionally substituted by one or more T.sup.2, C.sub.1-C.sub.3 fluoroalkyl optionally substituted by one or more T.sup.2, OC.sub.1-C.sub.3-fluoroalkyl optionally substituted by one or more T.sup.2, and (CH.sub.2).sub.p-(4- or 5- or 6-member heterocycle comprising at least one nitrogen atom that is aromatic or saturated or totally unsaturated or partially unsaturated) optionally substituted by one or more T.sup.2; T.sup.2, identical or different, is independently selected from the group consisting of OH, NH.sub.2, and CONH.sub.2; T, identical or different, is independently selected from the group consisting of F, C.sub.1-C.sub.3 alkyl, C.sub.1-C.sub.3 fluoroalkyl, OC.sub.1-C.sub.3 fluoroalkyl, -(L).sub.w-(CH.sub.2).sub.pC.sub.3-C.sub.6 cycloalkyl; -(L).sub.w-(CH.sub.2).sub.pC.sub.3-C.sub.6 cyclofluoroalkyl, -(L).sub.w-(CH.sub.2).sub.p-heterocycle, -(L).sub.w-(CH.sub.2).sub.pCN, -(L).sub.w-(CH.sub.2).sub.pOC(O)Q.sup.5, -(L).sub.w-(CH.sub.2).sub.pC(O)OQ.sup.5, -(L).sub.w-(CH.sub.2).sub.pOC(O)OQ.sup.5, -(L).sub.w-(CH.sub.2).sub.pOC(O)NQ.sup.5Q.sup.6, -(L).sub.w-(CH.sub.2).sub.pC(O)NQ.sup.5Q.sup.6, -(L).sub.w-(CH.sub.2).sub.pC(O)NQ.sup.5OQ.sup.6, -(L).sub.w-(CH.sub.2).sub.pC(O)NQ.sup.5NQ.sup.5Q.sup.6, -(L).sub.w-(CH.sub.2).sub.pNQ.sup.5C(O)Q.sup.6, -(L).sub.w-(CH.sub.2).sub.pNQ.sup.5Q(O).sub.2Q.sup.6, -(L).sub.w-(CH.sub.2).sub.pNQ.sup.5C(O)OQ.sup.6, -(L).sub.w-(CH.sub.2).sub.pNQ.sup.5C(O)NQ.sup.5Q.sup.6, -(L).sub.w-(CH.sub.2).sub.pNQ.sup.5Q.sup.6, -(L).sub.w-(CH.sub.2).sub.pNHC(NHQ.sup.3)NQ.sup.4, -(L).sub.w-(CH.sub.2).sub.pNHCHNQ.sup.3, and -(L).sub.w(CH.sub.2).sub.pC(NHQ.sup.3)NQ.sup.4; L, identical or different, is independently selected from the group consisting of O, S, N(R.sup.2), S(O), and S(O).sub.2; m is 1 or 2; n is 0, 1, or 2; p, identical or different, is independently selected from the group consisting of 0, 1, 2, and 3; q, identical or different, is independently selected from the group consisting of 2 and 3; v, identical or different, is independently selected from the group consisting of 1, 2, and 3; w, identical or different, is independently selected from the group consisting of 0 and 1; wherein any carbon atom present within any of the foregoing alkyls, cycloalkyls, fluoroalkyls, cyclofluoroalkyls, and heterocycles can be oxidized to form a CO group; wherein any sulphur atom present within a heterocycle can be oxidized to form a SO group or a S(O)2 group; and wherein any nitrogen atom present within a heterocycle or a tertiary amino group can be further quaternized by a methyl group; or a pharmaceutically acceptable salt, a zwitterion, a racemate, a diastereoisomer, an enantiomer, a geometric isomer, or a tautomer of formula (I).

2. The compound according to claim 1 selected from the group consisting of compounds of formulae (A), (B), (I*), (A*), and (B*) ##STR00046##

3. The compound according to claim 1 selected from group consisting of compounds of formulae (A1) to (A68) and (B1) to (B8) ##STR00047## ##STR00048## ##STR00049## ##STR00050## ##STR00051## ##STR00052## ##STR00053## ##STR00054## ##STR00055## ##STR00056## ##STR00057## ##STR00058## ##STR00059## ##STR00060## ##STR00061##

4. The compound according to claim 1, wherein R.sup.1 is one of the following: R.sup.1 is selected from the group consisting of a carbon-linked 4-, 5- or 6-membered heterocycle comprising at least one nitrogen atom that is aromatic or saturated or totally unsaturated or partially unsaturated and optionally substituted by one or more T.sup.1, H,CN; C(O)NHQ.sup.1; C(O)NHOQ.sup.1; C(O)NHNHQ.sup.1; (CH.sub.2)OQ.sup.1; and C(O)OQ.sup.1; or R.sup.1 is selected from the group consisting of (CH.sub.2)NHQ.sup.3 and (CH.sub.2)NHC(NHQ.sup.3)NQ.sup.4.

5. The compound according to claim 1, wherein R.sup.1 is one of the following: R.sup.1 is selected from the group consisting of CN; C(O)NHQ.sup.1, C(O)NHOQ.sup.1, and C(O)NHNHQ.sup.1; or R.sup.1 is selected from the group consisting of (CH.sub.2).sub.mOQ.sup.1, (CH.sub.2).sub.mNHC(O)Q.sup.1, (CH.sub.2).sub.mNHC(O)OQ.sup.1, and (CH.sub.2).sub.mNHC(O)NQ.sup.1Q.sup.2; or R.sup.1 is a carbon-linked 4- or 5- or 6-member heterocycle comprising at least one nitrogen atom that is saturated or partially unsaturated or totally unsaturated or aromatic and optionally substituted by one or more T.sup.1 and further comprises one or two or three additional heteroatoms wherein each additional heteroatom is selected from the group consisting of N, O, S, S(O), and S(O).sub.2; or R.sup.1 is H.

6. The compound according to claim 1, wherein: R.sup.2 is selected from the group consisting of (CH.sub.2).sub.qNQ.sup.5Q.sup.6, C(O)(CH.sub.2).sub.vNQ.sup.5Q.sup.6, (CH.sub.2).sub.qNHC(NHQ.sup.3)NQ.sup.4, C(O)NQ.sup.5Q.sup.6, (C(O)).sub.w(CH.sub.2).sub.vC(NHQ.sup.3)NQ.sup.4, C(NHQ.sup.3)NQ.sup.4, (C(O)).sub.w(CH.sub.2).sub.vC(O)NQ.sup.5Q.sup.6, (C(O)).sub.w(CH.sub.2).sub.p-(4- or 5- or 6-member heterocycle that is aromatic or saturated or totally unsaturated or partially unsaturated), (CH.sub.2).sub.qNHS(O).sub.2NQ.sup.5Q.sup.6; C(O)(CH.sub.2).sub.vNHS(O).sub.2NQ.sup.5Q.sup.6; (CH.sub.2).sub.qNHC(O)NQ.sup.5Q.sup.6; C(O)(CH.sub.2).sub.vNHC(O)NQ.sup.5Q.sup.6; (C(O)).sub.w(CH.sub.2).sub.vC(O)OQ.sup.5; (C(O)).sub.wC.sub.1-C.sub.3 alkyl; (CH.sub.2).sub.qNHC(O)OQ.sup.5; C(O)(CH.sub.2).sub.vNHC(O)OQ.sup.5; (CH.sub.2).sub.qOQ.sup.5, and C(O)(CH.sub.2).sub.vOQ.sup.5; and Q.sup.5 and Q.sup.6, identical or different, are independently selected from the group consisting of H, (CH.sub.2).sub.q, NHQ.sup.3, and C.sub.1-C.sub.4-alkyl.

7. The compound according to claim 1, wherein: R.sup.2 is selected from the group consisting of (CH.sub.2).sub.p-(4- or 5- or 6-member heterocycle that is aromatic or saturated or totally unsaturated or partially unsaturated), C.sub.1-C.sub.3 alkyl, (CH.sub.2).sub.vC(O)OQ.sup.5; (CH.sub.2).sub.qNHC(O)OQ.sup.5; (CH.sub.2).sub.qNQ.sup.5Q.sup.6, (CH.sub.2).sub.qOQ.sup.5, and (CH.sub.2).sub.vC(O)NH(CH.sub.2).sub.qNHQ.sup.3; Q.sup.5 and Q.sup.6, identical or different, are independently selected from the group consisting of H and C.sub.1-C.sub.4alkyl; and Q.sup.3 is H.

8. The compound according to claim 1, wherein: R.sup.1 is one of the following: R.sup.1 is selected from the group consisting of H, a carbon-linked 4- or 5- or 6-member heterocycle comprising at least one nitrogen atom that is aromatic or saturated or totally unsaturated or partially unsaturated and optionally substituted by one or more T.sup.1, CN, C(O)NHQ.sup.1, C(O)NHOQ.sup.1, C(O)NHNHQ.sup.1; (CH.sub.2)OQ.sup.1, and C(O)OQ.sup.1; or R.sup.1 is selected from the group consisting of (CH.sub.2)NHQ.sup.3 and (CH.sub.2)NHC(NHQ.sup.3)NQ.sup.4; or R.sup.1 is selected from the group consisting of H, a carbon-linked 4- or 5- or 6-member heterocycle comprising at least one nitrogen atom that is aromatic or saturated or totally unsaturated or partially unsaturated and optionally substituted by one or more T.sup.1, CN; C(O)NHQ.sup.1, C(O)NHOQ.sup.1, C(O)NHNHQ.sup.1, C(O)OQ.sup.1, (CH.sub.2)OQ.sup.1, (CH.sub.2)NHQ.sup.3, (CH.sub.2).sub.2NHQ.sup.3, (CH.sub.2)NHC(NHQ.sup.3)NQ.sup.4, (CH.sub.2).sub.2NHC(NHQ.sup.3)NQ.sup.4, (CH.sub.2)NHCHNQ.sup.3, (CH.sub.2).sub.2NHCHNQ.sup.3, and C(NHQ.sup.3)NQ.sup.4, Q.sup.1 is H or methyl; Q.sup.3 and Q.sup.4 are H; R.sup.3 is SO.sub.3H; W, unsubstituted or substituted by one or more T, is a non-aromatic, unsaturated 5-member heterocycle comprising at least one NR.sup.2 group and a (X).sub.n group; R.sup.2 is selected from the group consisting of (C(O)).sub.wC.sub.1-C.sub.3alkyl, (C(O)).sub.w(CH.sub.2).sub.vC(O)OQ.sup.5, (CH.sub.2).sub.qNHC(NHQ.sup.3)NQ.sup.4, (C(O)).sub.w(CH.sub.2).sub.vC(O)NQ.sup.5Q.sup.6, (C(O)).sub.w(CH.sub.2).sub.qNQ.sup.5Q.sup.6, C(O)(CH.sub.2).sub.vNHC(O)NQ.sup.5Q.sup.6, (C(O)).sub.w(CH.sub.2).sub.qOQ.sup.5, (C(O)).sub.w(CH.sub.2).sub.qNHC(O)OQ.sup.5, and (C(O)).sub.w(CH.sub.2).sub.p-(4- or 5- or 6-member heterocycle that is aromatic or saturated or totally unsaturated or partially unsaturated); Q.sup.5 and Q.sup.6, identical or different, are H or C.sub.1-C.sub.4alkyl; w is 0; and X is S.

9. The compound according to claim 1, wherein: R.sup.1 is H; R.sup.3 is SO.sub.3H; W, unsubstituted or substituted by one or more T, is a non-aromatic, unsaturated 5-member heterocycle comprising at least one NR.sup.2 group and a (X).sub.n group; R.sup.2 is selected from the group consisting of (CH.sub.2).sub.p-(4- or 5- or 6-member heterocycle that is aromatic or saturated or totally unsaturated or partially unsaturated), C.sub.1-C.sub.3 alkyl, (CH.sub.2).sub.vC(O)OQ.sup.5, (CH.sub.2).sub.qNHC(O)OQ.sup.5, (CH.sub.2).sub.qNQ.sup.5Q.sup.6, (CH.sub.2).sub.qOQ.sup.5, and (CH.sub.2).sub.qC(O)NQ.sup.5Q.sup.6; Q.sup.5 and Q.sup.6, identical or different, are H or C.sub.1-C.sub.4alkyl; and X is S.

10. The compound according to claim 1 selected from the group consisting of ##STR00062## wherein PG, is a protective group.

11. The compound according to claim 1 selected from the group consisting of ##STR00063## wherein PG is a protective group selected from the group consisting of allyl, benzyl, tertbutyldimethylsilyl (TBDMS), and tert-butoxycarbonyl (Boc).

12. A pharmaceutical composition comprising at least one compound according to claim 1.

13. The pharmaceutical composition according to claim 12 further comprising a an antibacterial compound selected from the group consisting of selected from aminoglycosides, -lactams, glycylcyclines, tetracyclines, quinolones, fluoroquinolones, glycopeptides, lipopeptides, macrolides, ketolides, lincosamides, streptogramins, oxazolidinones, polymyxins and combinations thereof.

14. The pharmaceutical composition according to claim 12, wherein the antibacterial compound is -lactams selected from the group consisting penicillin, cephalosporins, penems, carbapenems, monobactam, and combinations thereof.

15. The pharmaceutical composition according to claim 12 further comprising ceftazidime.

16. The pharmaceutical composition according to claim 12 further comprising a pharmaceutically acceptable excipient.

17. A kit comprising a pharmaceutical composition according to claim 11 and at least one second composition according to claim 11.

18. The kit according to claim 17 further comprising ceftazidime.

19. A method for the treatment of bacterial infections comprising the administration of a therapeutically effective amount of the compound of claim 1.

20. The method according to claim 19, wherein the bacterial infection is caused by bacteria producing one or more beta-lactamases.

21. A method according to claim 19, wherein the bacterial infection is caused by gram-negative bacteria.

Description

EXAMPLES

(1) The following examples are provided for the purpose of illustrating the present invention and by no means should be interpreted to limit the scope of the present invention.

(2) The first part represents the preparation of the compounds according to the invention (intermediates and final compounds) whereas the second part describes the evaluation of antibacterial activity of compounds according to the invention.

(3) Preparation of the Compounds and Biological Activity:

(4) Abbreviations or symbols used herein include: ACHN: 1,1-azobis(cyclohexanecarbonitrile) ACN: acetonitrile AcOH: acetic acid Bn: benzyl Boc: tert-butoxycarbonyl Boc.sub.2O: tert-butoxycarbonyl anhydride BocON: [2-(tert-butoxycarbonyloxyimino)-2-phenylacetonitrile] bs: broad singlet Burgess reagent: methyl N-(triethylammoniosulfonyl)carbamate 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 dq: doublet of quartet dt: doublet of triplet DTA: di-tert-butylazodicarboxylate DEAD: diethyl azodicarboxylate Dess-Martin periodinane: 1,1,1-tris(acetyloxy)-1,1-dihydro-1,2-benziodoxol-3-(1H)-one DIAD: diisopropyl azodicarboxylate DIPEA: N,N-diisopropylethylamine DMAP: 4-dimethylaminopyridine DMF: N,N-dimethylformamide 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 m: multiplet min: minutes MeOH: methanol MeONa: sodium methoxide MIC: minimum inhibitory concentration MS: mass spectrometry MsCl: methanesulfonyl chloride NBS: N-bromosuccinimide NMR: nuclear magnetic resonance spectroscopy Ns: nosyl, nitrobenzenesulfonyl Pd(Ph.sub.3).sub.4: tetrakis(triphenylphosphine)palladium(0) PG: protective group PhSH: thiophenol PMe.sub.3: trimethylphosphine PPh.sub.3: triphenylphosphine Ppm: parts per million q: quartet rt: room temperature s: singlet SEM: [2-(trimethylsilyl)ethoxy]methyl t: triplet td: triplet of doublet TBAF: tetra-n-butylammonium fluoride TBDMSOTf: trifluoromethanesulfonic acid tert-butyldimethylsilyl ester TBSOTf: trimethylsilyl trifluoromethanesulfonate tBuOK: potassium tert-butoxide TEA: trimethylamine TFA: trifluoroacetic acid THF: tetrahydrofuran THP: tetrahydropyranyl TLC: thin layer chromatography TMSI: lodotrimethylsilane Tr: trityl (triphenylmethyl)

Example 1

Synthesis of Sodium (5-methyl-4,9-dioxo-3-thia-5,8,10-triaza tricyclo[6.2.1.02,6]undec-2(6)-en-10-yl) sulfate

(5) ##STR00034## ##STR00035##

Step 1: Preparation of Intermediate tert-butyl 4-bromo-3,5-dioxo-piperidine-1-carboxylate (1a)

(6) To a solution of tert-butyl 3,5-dioxopiperidine-1-carboxylate (3 g, 14.07 mmol) in anhydrous DCM (60 mL) under inert atmosphere at 0 C. was successively added NBS (2.5 g, 14.07 mmol) and ACHN (0.223 g, 0.91 mmol). The reaction mixture was stirred 2 h at 0 C. The solution was washed with water, then with NaCl aqueous solution. The organic layer was dried over Na.sub.2SO.sub.4 and evaporated in vacuo to give tert-butyl 4-bromo-3,5-dioxo-piperidine-1-carboxylate (1a) (4.11 g, 14.07 mmol, quantitative yield) as an off-white solid.

(7) MS m/z ([M+H-tertbutyl].sup.+) 236/238.

(8) MS m/z ([MH].sup.) 290/292.

(9) .sup.1H NMR (400 MHz, CDCl.sub.3): (ppm) 1.48 (s, 9H), 4.35 (bs, 4H).

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

(10) To a solution of tert-butyl 4-bromo-3,5-dioxo-piperidine-1-carboxylate (1a) (0.500 g, 1.71 mmol) in anhydrous MeOH (8 mL) under inert atmosphere was added thiourea (0.226 g, 3.42 mmol). After stirring 30 min at rt, TEA (0.477 mL, 3.42 mmol) was added and the mixture was refluxed 5 h. MeOH was evaporated and the residue was solubilized with EtOAc. The solution was washed with water, 10% of Na.sub.2CO.sub.3 aqueous solution and NaCl aqueous solution. The organic phase was dried over Na.sub.2SO.sub.4, filtered and evaporated. The solid was triturated with cyclohexane and filtered to give tert-butyl 2-amino-7-oxo-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-5-carboxylate (1b) (0.350 g, 1.30 mmol, 76%) as an off white solid.

(11) MS m/z ([M+H].sup.+) 270.

(12) MS m/z ([MH].sup.31 ) 268.

(13) .sup.1H NMR (400 MHz, CDCl.sub.3): (ppm) 1.48 (s, 9H), 4.23 (s, 2H), 4.66 (s, 2H), 5.77 (bs, 2H).

Step 3: Preparation of Intermediate tert-butyl 2-chloro-7-oxo-4,6-dihydrothiazolo[4,5-c]pyridine-5-carboxylate (1c)

(14) To a solution of tert-butyl 2-amino-7-oxo-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-5-carboxylate (1b) (0.473 g, 1.76 mmol) in anhydrous ACN (33 mL) under inert atmosphere at 20 C. was added isoamyl nitrite (0.710 mL, 5.27 mmol). After 10 min at 20 C., Copper(11) chloride (0.473 g, 3.52 mmol) was added. The mixture was stirred for 1 h at 20 C., then 4 h at rt. The solution was extracted with DCM, washed with 10% of Na.sub.2CO.sub.3 aqueous solution. The organic phase was dried over Na.sub.2SO.sub.4, filtered and evaporated. The product was purified by flash chromatography on silica gel (DCM/EtOAc 98/2) to give tert-butyl 2-chloro-7-oxo-4,6-dihydrothiazolo[4,5-c]pyridine-5-carboxylate (1c) (0.458 g, 1.59 mmol, 90%) as an off-white solid.

(15) MS m/z ([M+H-tertbutyl].sup.+) 233/235.

(16) MS m/z ([MH].sup.) 287/289.

(17) .sup.1H NMR (400 MHz, CDCl.sub.3): (ppm) 1.48 (s, 9H), 4.31 (s, 2H), 4.83 (s, 2H).

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

(18) To a solution of tert-butyl 2-chloro-7-oxo-4,6-dihydrothiazolo[4,5-c]pyridine-5-carboxylate (1c) (3.75 g, 12.98 mmol) in anhydrous MeOH (97 mL) under inert atmosphere at 78 C. was added dropwise a MeONa solution 0.5M (28.6 mL, 14.30 mmol). The reaction mixture was stirred for 15 min at 78 C., then for 30 min at rt. MeOH was removed under vacuum and the resulting residue was diluted with DCM and filtered on a mixture of silica gel and celite. The product was eluted with DCM/EtOAc 8/2, concentrated in vacuo and purified by flash chromatography on silica gel (cyclohexane/EtOAc 90/10 to 80/20) to provide tert-butyl 2-methoxy-7-oxo-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-5-carboxylate (1d) (2.23 g, 7.84 mmol, 60%) as a yellow solid.

(19) MS m/z ([M+H].sup.+) 285.

(20) .sup.1H NMR (400 MHz, CDCl.sub.3): (ppm) 1.49 (s, 9H), 4.17 (s, 3H), 4.26 (s, 2H), 4.70 (s, 2H).

Step 5: Preparation of Intermediate tert-butyl 3-methyl-2,7-dioxo-2,3,6,7-tetrahydro-4H-thiazolo[4,5-c]pyridine-5-carboxylate (1e)

(21) Tert-butyl 2-methoxy-7-oxo-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-5-carboxylate (1d) (1.92 g, 6.74 mmol) was solubilized in pyridine (1.1 mL, 13.49 mmol) and warmed 30 min at 90 C. under microwave irradiation. The mixture was evaporated and the residue was diluted with DCM (57 mL) at 0 C. A solution of Mel (0.840 mL, 13.49 mmol) in DCM (15 mL) was added dropwise to the reaction mixture. After stirring 1 h at 0 C. and 2 h at rt, the precipitate was filtered. The filtrate was evaporated, solubilized in EtOAc and filtrated on silica gel cake to provide tert-butyl 3-methyl-2,7-dioxo-2,3,6,7-tetrahydro-4H-thiazolo[4,5-c]pyridine-5-carboxylate (1e) (1.63 g, 5.72 mmol, 85%).

(22) MS m/z ([M+H].sup.+) 285.

(23) MS m/z ([MH].sup.) 283.

(24) .sup.1H NMR (400 MHz, CDCl.sub.3): (ppm) 1.49 (s, 9H), 3.36 (s, 3H), 4.26 (s, 2H), 4.61 (s, 2H).

Step 6: Preparation of Intermediate tert-butyl 7-hydroxy-3-methyl-2-oxo-2,3,6,7-tetrahydro-4H-thiazolo[4,5-c]pyridine-5-carboxylate (1f)

(25) To a solution of tert-butyl 3-methyl-2,7-dioxo-2,3,6,7-tetrahydro-4H-thiazolo[4,5-c]pyridine-5-carboxylate (1e) (1.78 g, 6.25 mmol) in anhydrous MeOH (62 mL) at 0 C. under inert atmosphere was added sodium borohydride (0.236 g, 6.25 mmol) by portions. The reaction mixture was stirred for 1 h and then concentrated under vacuum. The residue was diluted with EtOAc and filtered on silica gel cake to provide tert-butyl 7-hydroxy-3-methyl-2-oxo-2,3,6,7-tetrahydro-4H-thiazolo[4,5-c]pyridine-5-carboxylate (1f) (1.56 g, 5.45 mmol, 87%) as a yellow solid.

(26) MS m/z ([M+H].sup.+) 287.

(27) .sup.1H NMR (400 MHz, CDCl.sub.3): (ppm) 1.50 (s, 9H), 3.22 (s, 3H), 3.55 (dd, J=3.0/13.6 Hz, 1H), 3.97-4.01 (m, 1H), 4.04 (bs, 1H), 4.52 (bs, 2H).

Step 7: Preparation of Intermediate tert-butyl 7-[allyloxy-(2-nitro-benzenesulfonyl)-amino]-3-methyl-2-oxo-2,3,6,7-tetrahydro-4H-thiazolo[4,5-c]pyridine-5-carboxylate (1g)

(28) To a solution of tert-butyl 7-hydroxy-3-methyl-2-oxo-2,3,6,7-tetrahydro-4H-thiazolo[4,5-c]pyridine-5-carboxylate (1f) (1.56 g, 5.45 mmol) in anhydrous toluene (60 mL) under inert atmosphere was added N-allyloxy-2-nitro-benzenesulfonamide (1.41 g, 5.45 mmol) and Ph.sub.3P (1.43 g, 5.45 mmol). DTA (1.42 g, 6.16 mmol) was added by portion and the mixture was stirred for 4 h at rt. The solution was filtered and concentrated under vacuum. The residue was purified by flash chromatography on silica gel (DCM/cyclohexane 70/30 to DCM/MeOH 99/1) to give tert-butyl 7-[allyloxy-(2-nitro-benzenesulfonyl)-amino]-3-methyl-2-oxo-2,3,6,7-tetrahydro-4H-thiazolo[4,5-c]pyridine-5-carboxylate (1g) (2.39 g, 4.53 mmol, 83%) as a yellow solid.

(29) MS m/z ([M+H].sup.+) 527.

Step 8: Preparation of Intermediate tert-butyl 7-allyloxyamino-3-methyl-2-oxo-2,3,6,7-tetrahydro-4H-thiazolo[4,5-c]pyridine-5-carboxylate (1h)

(30) To a solution of tert-butyl 7-[allyloxy-(2-nitro-benzenesulfonyl)-amino]-3-methyl-2-oxo-2,3,6,7-tetrahydro-4H-thiazolo[4,5-c]pyridine-5-carboxylate (1g) (0.500 g, 0.95 mmol) in anhydrous ACN (6 mL) under inert atmosphere was added successively PhSH (0.487 mL, 4.75 mmol) and K.sub.2CO.sub.3 (0.985 g, 7.12 mmol). The reaction mixture was stirred for 16 h at rt. The reaction mixture was concentrated under vacuum, diluted with DCM and filtered to eliminate salts. The residue was purified by flash chromatography on silica gel (DCM/MeOH 100/0 to 95/5) then purified by preparative TLC (DCM/MeOH 95/5) to provide tert-butyl 7-allyloxyamino-3-methyl-2-oxo-2,3,6,7-tetrahydro-4H-thiazolo[4,5-c]pyridine-5-carboxylate (1h) (0.289 g, 0.84 mmol, 89%).

(31) MS m/z ([M+H].sup.+) 342.

(32) .sup.1H NMR (400 MHz, CDCl.sub.3): (ppm) 1.47 (s, 9H), 3.21 (s, 3H), 3.30-3.40 (m, 1H), 3.81-4.06 (m, 2H), 4.17-4.26 (m, 2H), 4.35-4.69 (m, 1H), 5.16-5.32 (m, 2H), 5.38-5.43 (m, 1H), 5.87-5.97 (m, 1H), 6.21 (bs, 1H).

Step 9: Preparation of Intermediate 7-allyloxyamino-3-methyl-4,5,6,7-tetrahydro-3H-thiazolo[4,5-c]pyridin-2-one (1i)

(33) To a solution of 7-allyloxyamino-3-methyl-2-oxo-2,3,6,7-tetrahydro-4H-thiazolo[4,5-c]pyridine-5-carboxylic acid tert-butyl ester (1h) (0.289 g, 0.85 mmol) in anhydrous DCM (5 mL) under inert atmosphere was added TFA (1 mL, 12.7 mmol). After stirring for 2 h at rt, the solution was cooled at 0 C. and neutralized to pH 8 with NH.sub.4OH solution 28%. The solution was diluted with water, the organic layer was separated from the aqueous, dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuo to 7-allyloxyamino-3-methyl-4,5,6,7-tetrahydro-3H-thiazolo[4,5-c]pyridin-2-one (1i) (0.140 g, 0.58 mmol, 68%) as a yellow oil.

(34) .sup.1H NMR (400 MHz, CDCl.sub.3): (ppm) 2.97 (dd, J=3.6/13.6 Hz, 1H), 3.17 (s, 3H), 3.30 (dd, J=2.8/13.6 Hz, 1H), 3.59 (d, J=1.5/16.5 Hz, 1H), 3.69 (d, J=1.0/16.3 Hz, 1H), 3.75 (bs, 1H), 4.20 (dt, J=1.2/6.0 Hz, 2H), 5.20-5.25 (m, 2H), 5.61 (bs, 1H), 5.88-5.97 (m, 1H).

Step 10: Preparation of Intermediate 10-allyloxy-5-methyl-3-thia-5,8,10-triaza-tricyclo[6.2.1.02.6]undec-2(6)-ene-4,9-dione (1j)

(35) To a solution of 7-allyloxyamino-3-methyl-4,5,6,7-tetrahydro-3H-thiazolo[4,5-c]pyridin-2-one (1i) (0.140 g, 0.58 mmol) in anhydrous ACN (90 mL) at 10 C. under inert atmosphere was added TEA (0.323 mL, 2.32 mmol). A solution of diphosgene (0.035 mL, 0.29 mmol) in ACN (11 mL) was added dropwise at 10 C. After 2 h at 10 C. then 18 h at rt, the mixture reaction was bubbled 30 min under nitrogen and concentrated under vacuum. The residue was diluted with DCM, washed with water and NaCl aqueous solution. The organic layer was dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The product was purified by preparative TLC (DCM/MeOH 96/4) to provide 10-allyloxy-5-methyl-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2.6] undec-2(6)-ene-4,9-dione (1j) (0.045 g, 0.17 mmol, 29%) as an colourless oil.

(36) MS m/z ([M+H].sup.+) 268.

(37) .sup.1H NMR (400 MHz, CDCl.sub.3): (ppm) 3.16 (s, 3H), 3.28 (dd, J=0.4/11 Hz, 1H), 3.70 (dd, J=3.2/11 Hz, 1H), 4.06 (d, J=16.0 Hz, 1H), 4.20-4.24 (m, 2H), 4.39-4.51 (m, 2H), 5.32-5.38 (m, 2H), 5.97-6.07 (m, 1H).

Step 11: Preparation of Intermediate 10-hydroxy-5-methyl-3-thia-5,8,10-triaza-tricyclo[6.2.1.02.6]undec-2(6)-ene-4,9-dione (1k)

(38) To a solution of 10-allyloxy-5-methyl-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2.6]undec-2(6)-ene-4,9-dione (1j) (0.050 g, 0.19 mmol) and glacial AcOH (0.011 mL, 0.19 mmol) in anhydrous DCM (3.2 mL) was added in one portion Pd(PPh.sub.3).sub.4 (0.108 g, 0.09 mmol). After stirring for 3 h at rt, the mixture was concentrated and purified by flash chromatography (DCM/acetone 100/0 to 80/20) to provide 10-hydroxy-5-methyl-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2.6]undec-2(6)-ene-4,9-dione (1k) (0.022 g, 0.10 mmol, 51%).

(39) MS m/z ([M+H].sup.+) 228.

(40) .sup.1H NMR (300 MHz, CDCl.sub.3): (ppm) 3.13 (s, 3H), 3.24 (d, J=10.8 Hz, 1H), 3.66 (dd, J=2.9/10.9 Hz, 1H), 4.01 (d, J=16.7 Hz, 1H), 4.15 (s, 1H), 4.18 (d, J=16.8 Hz, 1H).

Step 12: Preparation of Sodium (5-methyl-4,9-dioxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.02.6]undec-2(6)-en-10-yl) sulfate (Example 1)

(41) To a solution of 10-hydroxy-5-methyl-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2.6]undec-2(6)-ene-4,9-dione (1k) (0.022 g, 0.10 mmol) in anhydrous DCM (1.5 mL) was added a suspension of sulfur trioxide pyridine complex (0.092 mg, 0.58 mmol) in dry pyridine (1 mL) and the resulting solution was protected from light and stirred overnight at rt until the sulfatation was completed. The reaction mixture was concentrated under vacuum, diluted with DCM and filtered. The filtrate was co-evaporated with toluene and dried under vacuum. The residue was solubilized in a minimal volume of water and 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, frozen and lyophilized to afford sodium (5-methyl-4,9-dioxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2.6]undec-2(6)-en-10-yl) sulfate (Example 1) (0.032 g, 0.10 mmol, quantitative yield) as a white solid.

(42) MS m/z ([MH].sup.) 306.

(43) .sup.1H NMR (300 MHz, D.sub.2O): (ppm) 3.20 (s, 3H), 3.58 (d, J=11.5 Hz, 1H), 3.84 (dd, J=3.0/11.5 Hz, 1H), 4.29 (d, J=16.9 Hz, 1H), 4.40 (d, J=16.9 Hz, 1H), 4.80 (bs, in D.sub.2O peak, 1H).

Example 2

Synthesis of Sodium [5-(2-methoxy-2-oxo-ethyl)-4,9-dioxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.02.6]undec-2(6)-en-10-yl] sulfate

(44) ##STR00036## ##STR00037##

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

(45) To a solution of tert-butyl 2-methoxy-7-oxo-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-5-carboxylate (1d) (7.19 g, 25.31 mmol) in anhydrous dioxane (193 mL) was added drop by drop HCl 12 N (2.50 mL). The reaction mixture was stirred for 4 h at 70 C. then concentrated under vacuum. The residue was diluted with EtOAc and washed with water. The organic layer was dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuo to provide tert-butyl 2,7-dioxo-4,6-dihydro-3H-thiazolo[4,5-c]pyridine-5-carboxylate (2a) (3.43 g, 12.70 mmol, 50%) as an orange solid.

(46) MS m/z ([MtBu+H].sup.+) 215.

(47) MS m/z ([MH].sup.) 269.

Step 2: Preparation of Intermediate tert-butyl 3-(2-methoxy-2-oxo-ethyl)-2,7-dioxo-4,6-dihydrothiazolo[4,5-c]pyridine-5-carboxylate (2b)

(48) To a solution of tert-butyl 2,7-dioxo-4,6-dihydro-3H-thiazolo[4,5-c]pyridine-5-carboxylate (2a) (418 mg, 1.55 mmol) in anhydrous acetone (15 mL) under inert atmosphere were added K.sub.2CO.sub.3 (214 mg, 1.55 mmol) and methyl-2-bromoacetate (146 L, 1.55 mmol) and the mixture was stirred for 2 h at 55 C. and concentrated in vacuo. The residue was diluted with EtOAc, washed with NaCl aqueous solution, dried over Na.sub.2SO.sub.4, filtered and concentrated. The crude was purified by flash chromatography on silica gel (DCM/MeOH 98/2) to provide tert-butyl 3-(2-methoxy-2-oxo-ethyl)-2,7-dioxo-4,6-dihydrothiazolo[4,5-c]pyridine-5-carboxylate (2b) (493 mg, 1.44 mmol, 93%) as a yellow oil.

(49) MS m/z ([MtBu+H].sup.+) 287.

(50) MS m/z ([MH].sup.) 341.

(51) .sup.1H NMR (400 MHz, CDCl.sub.3): (ppm) 1.48 (s, 9H), 3.82 (s, 3H), 4.28 (s, 2H), 4.54 (s, 2H), 5.55 (s, 2H).

Step 3: Preparation of Intermediate tert-butyl 7-hydroxy-3-(2-methoxy-2-oxo-ethyl)-2-oxo-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-5-carboxylate (2c)

(52) Using the procedure described in example 1 (step 6), tert-butyl 3-(2-methoxy-2-oxo-ethyl)-2,7-dioxo-4,6-dihydrothiazolo[4,5-c]pyridine-5-carboxylate (2b) (570 mg, 1.66 mmol) was converted to tert-butyl 7-hydroxy-3-(2-methoxy-2-oxo-ethyl)-2-oxo-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-5-carboxylate (2c) (424 mg, 1.23 mmol, 74%) as a yellow oil.

(53) MS m/z ([M+Na].sup.+) 367.

(54) MS m/z ([MH].sup.) 343.

(55) .sup.1H NMR (400 MHz, CDCl.sub.3): (ppm) 1.49 (s, 9H), 3.57 (dd, J=3.4/13.8 Hz, 1H), 3.79 (s, 3H), 3.91-3.98 (m, 1H), 4.01 (dd, J=3.9/13.8 Hz, 1H), 4.29-4.59 (m, 4H).

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

(56) Using the procedure described in example 1 (step 7), tert-butyl 7-hydroxy-3-(2-methoxy-2-oxo-ethyl)-2-oxo-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-5-carboxylate (2c) (424 mg, 1.2 mmol) was converted to tert-butyl 7-[allyloxy-(2-nitrophenyl)sulfonyl-amino]-3-(2-methoxy-2-oxo-ethyl)-2-oxo-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-5-carboxylate (2d) (257 mg, 0.44 mmol, 35%) as a yellow oil after a purification by flash chromatography on silica gel (cyclohexane/EtOAc 6/4).

Step 5: Preparation of Intermediate tert-butyl 7-(allyloxyamino)-3-(2-methoxy-2-oxo-ethyl)-2-oxo-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-5-carboxylate (2e)

(57) Using the procedure described in example 1 (step 8), tert-butyl 7-[allyloxy-(2-nitrophenyl)sulfonyl-amino]-3-(2-methoxy-2-oxo-ethyl)-2-oxo-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-5-carboxylate (2d) (256 mg, 0.40 mmol) is converted to tert-butyl 7-(allyloxyamino)-3-(2-methoxy-2-oxo-ethyl)-2-oxo-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-5-carboxylate (2e) (118 mg, 0.30 mmol, 69%) after a purification by preparative TLC

(58) (DCM/MeOH 95/5).

(59) MS m/z ([M+H].sup.+) 400.

(60) .sup.1H NMR (400 MHz, CDCl.sub.3): (ppm) 1.44 (s, 9H), 3.31-3.42 (m, 1H), 3.73 (s, 3H), 3.84-3.96 (m, 2H), 4.19 (d, J=5.8 Hz, 2H), 4.25-4.29 (m, 1H), 4.34-4.52 (m, 2H), 5.14-5.17 (m, 1H), 5.22-5.29 (m, 2H), 5.83-5.93 (m, 1H).

Step 6: Preparation of Intermediate 10-allyloxy-5-(2-methoxy-2-oxo-ethyl)-3-thia-5,8,10-triaza-tricyclo[6.2.1.02.6]undec-2(6)-ene-4,9-dione (2f)

(61) To a solution of tert-butyl 7-(allyloxyamino)-3-(2-methoxy-2-oxo-ethyl)-2-oxo-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-5-carboxylate (2e) (1.5 g, 3.77 mmol) in anhydrous DCM (38 mL) was added drop by drop TFA (4.4 mL, 56.48 mmol). The mixture was stirred for 3 h at 0 C., then neutralized with an ammonium hydroxide 28% solution (pH 7-8) and extracted twice with DCM. The combined organic layers were dried over Na.sub.2SO.sub.4, filtered and concentrated.

(62) The crude was dissolved in ACN (627 mL) and cooled down at 10 C. TEA (2.1 mL, 15.06 mmol) and a solution of diphosgene (227 L, 0.1.88 mmol) in ACN (70 mL) were added. After 1 h at rt, the mixture was stirred at 45 C. and TEA was added until total consumption of starting material. The crude was concentrated in vacuo and purified by flash chromatography on silica gel (DCM/MeOH 96/4) to afford 10-allyloxy-5-(2-methoxy-2-oxo-ethyl)-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2.6]undec-2(6)-ene-4,9-dione (2f) (569 mg, 1.74 mmol, 51%).

(63) MS m/z ([M+H].sup.+) 326.

(64) 1H NMR (300 MHz, CDCl.sub.3): (ppm) 3.31 (d, J=10.7 Hz, 1H), 3.66 (dd, J=2.9/11.0 Hz, 1H), 3.76 (s, 3H), 3.97-4.11 (m, 3H), 4.19 (d, J=2.4 Hz, 1H), 4.34-4.48 (m, 2H), 4.60 (d, J=17.8 Hz, 1H), 5.27-5.38 (m, 2H), 5.91-6.04 (m, 1H).

Step 7: Preparation of Sodium [5-(2-methoxy-2-oxo-ethyl)-4,9-dioxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.02.6]undec-2(6)-en-10-yl] sulfate (example 2)

(65) To a solution of 10-allyloxy-5-(2-methoxy-2-oxo-ethyl)-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2.6]undec-2(6)-ene-4,9-dione (2f) (42.2 mg, 0.13 mmol) and glacial AcOH (15 L, 0.23 mmol) in anhydrous DCM (930 L) was added in one portion Pd(Ph.sub.3).sub.4 (75 mg, 0.07 mmol). After stirring for 30 min at rt, dry pyridine (772 L) and sulfur trioxide pyridine complex (103 mg, 0.65 mmol) were added to the mixture and the resulting solution was protected from light and stirring overnight at rt until sulfatation was completed. The reaction mixture was concentrated in vacuo, diluted with DCM and filtered. The filtrate was concentrated and purified on silica gel (DCM/acetone 100/0 to 60/40) to provide 38 mg of a uncolored oil of triphenyl-(propenyl)-phosphonium salt [5-(2-methoxy-2-oxo-ethyl)-4,9-dioxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2.6]undec-2(6)-en-10-yl]sulfate. This oil was solubilized in a minimal volume of water and ACN and 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, frozen and lyophilized to afford sodium [5-(2-methoxy-2-oxo-ethyl)-4,9-dioxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2.6]undec-2(6)-en-10-yl] sulfate (example 2) (11.4 mg, 0.029 mmol, 22%) as a white solid.

(66) MS m/z ([MH].sup.) 364.

(67) .sup.1H NMR (400 MHz, D.sub.2O): (ppm) 3.63 (d, J=11.5 Hz, 1H), 3.82 (s, 3H), 3.87 (dd, J=3.0/11.5 Hz, 1H), 4.20-4.36 (m, 2H), 4.52-4.62 (m, 2H), 4.85 (d, J=2.7 Hz, 1H).

Example 3

Synthesis of Sodium [5-(2-(tert-butoxycarbonylamino)ethyl)-4,9-dioxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.02.6]undec-2(6)-en-10-yl] sulfate

(68) ##STR00038## ##STR00039##

Step 1: Preparation of Intermediate tert-butyl 3-(2-hydroxyethyl)-2,7-dioxo-4,6-dihydrothiazolo[4,5-c]pyridine-5-carboxylate (3a)

(69) To a solution of tert-butyl 2,7-dioxo-4,6-dihydro-3H-thiazolo[4,5-c]pyridine-5-carboxylate (2a) (11 g, 40.70 mmol) in anhydrous ACN (90 mL) under inert atmosphere were added K.sub.2CO.sub.3 (7.3 g, 52.90 mmol) and iodoethanol (12.7 mL, 162 mmol) and the mixture was stirred for 7 h at 70 C. and concentrated in vacuo. The residue was diluted with EtOAc and water. The aqueous layer was acidified (pH 2) with 1N Hydrochloric acid solution and extracted with EtOAc twice. The organic layer were combined, washed with water and NaCl aqueous solution, dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuo to provide tert-butyl 3-(2-hydroxyethyl)-2,7-dioxo-4,6-dihydrothiazolo[4,5-c]pyridine-5-carboxylate (3a) (18 g) as a yellow oil. The crude was engaged in the next step without purification.

(70) MS m/z ([M+H].sup.+) 315.

(71) MS m/z ([MH].sup.) 313.

(72) .sup.1H NMR (400 MHz, DMSO): (ppm) 1.42 (s, 9H), 3.60 (t, J=4.9 Hz, 2H), 3.81 (t, J=4.9 Hz, 2H), 4.18 (s, 2H), 4.77 (bs, 2H).

Step 2: Preparation of Intermediate tert-butyl 3-[2-[tert-butyl(dimethyl)silyl]oxyethyl]-2,7-dioxo-4,6-dihydrothiazolo[4,5-c]pyridine-5-carboxylate (3b)

(73) To a solution of crude compound tert-butyl 3-(2-hydroxyethyl)-2,7-dioxo-4,6-dihydrothiazolo[4,5-c]pyridine-5-carboxylate (3a) (2.70 g, 8.64 mmol) in anhydrous DMF under inert atmosphere were added tert-butyl-chloro-dimethyl-silane (1.43 g, 9.50 mmol) and imidazole (1.17 g, 17.18 mmol). The mixture was stirred for 1 h at rt then ice and EtOAc were added. The organic layer was separated, washed with a saturated solution of NaCl, dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuo to provide tert-butyl 3-[2-[tert-butyl(dimethyl)silyl]oxyethyl]-2,7-dioxo-4,6-dihydrothiazolo[4,5-c]pyridine-5-carboxylate (3b) (2.46 g, 5.74 mmol, 66% over 2 steps) as a brown oil which was engaged in the next step without further purification.

(74) MS m/z ([M+H].sup.+) 429.

(75) MS m/z ([MH].sup.) 427.

(76) .sup.1H NMR (400 MHz, CDCl.sub.3): (ppm) 0.00 (s, 6H), 0.83 (s, 9H), 1.49 (s, 9H), 3.87 (s, 4H), 4.23 (s, 2H), 4.68 (bs, 2H).

Step 3: Preparation of Intermediate tert-butyl 3-[2-[tert-butyl(dimethyl)silyl]oxyethyl]-7-hydroxy-2-oxo-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-5-carboxylate (3c)

(77) Using the procedure described in example 1 (step 6), tert-butyl 3-[2[tert-butyl(dimethyl)silyl]oxyethyl]-2,7-dioxo-4,6-dihydrothiazolo[4,5-c]pyridine-5-carboxylate (3b) (15.09 g, 35.20 mmol) is converted to tert-butyl 3-[2-[tert-butyl(dimethyl)silyl]oxyethyl]-7-hydroxy-2-oxo-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-5-carboxylate (3c) (6.76 g, 15.6 mmol, 44%) as a yellow oil after a purification by flash chromatography on silica gel (DCM/MeOH 100/0 to 98/2).

(78) MS m/z ([M+H].sup.+) 431.

(79) .sup.1H NMR (400 MHz, CDCl.sub.3): (ppm) 0.00 (bs, 6H), 0.84 (s, 9H), 1.50 (s, 9H), 3.48 (dd, J=3.4/13.7 Hz, 1H), 3.66-3.84 (m, 4H), 4.00-4.10 (m, 2H), 4.50-4.73 (m, 2H)

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

(80) Using the procedure described in example 1 (step 7), tert-butyl 3-[2-[tert-butyl(dimethyl)silyl]oxyethyl]-7-hydroxy-2-oxo-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-5-carboxylate (3c) (6.70 g, 15.56 mmol) is converted to tert-butyl 7-[allyloxy-(2-nitrophenyl)sulfonyl-amino]-3-[2-[tert-butyl(dimethyl)silyl]oxyethyl]-2-oxo-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-5-carboxylate (3d) using N-allyloxy-2-nitro-benzenesulfonamide (4.82 g, 18.67 mmol), PPh.sub.3 (4.08 g, 18.67 mmol) and DTA (4.30 g, 18.6 mmol). After 1 h of reaction, magnesium chloride (3.60 g, 37.30 mmol) was added and the reaction mixture was stirred for 2 h at 60 C. then overnight at rt. The solution was filtered and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (toluene/acetone 100/0 to 90/10) to provide compound (3d) (5.54 g, 8.20 mmol, 53%) as a yellow oil.

Step 5: Preparation of Intermediate tert-butyl 7-(allyloxyamino)-3-[2-[tert-butyl(dimethyl)silyl]oxyethyl]-2-oxo-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-5-carboxylate (3e)

(81) Using the procedure described in example 1 (step 8), tert-butyl 7-[allyloxy-(2-nitrophenyl)sulfonyl-amino]-3-[2-[tert-butyl(dimethyl)silyl]oxyethyl]-2-oxo-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-5-carboxylate (3d) (5.2 g, 7.76 mmol) is converted to tert-butyl 7-(allyloxyamino)-3-[2-[tert-butyl(dimethyl)silyl]oxyethyl]-2-oxo-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-5-carboxylate (3e) (2.53 g, 5.21 mmol, 67%) after a purification on silica gel (cyclohexane/EtOAc 80/20 to 60/40).

Step 6: Preparation of Intermediate 10-Allyloxy-5-(2-hydroxy-ethyl)-3-thia-5,8,10-triaza-tricyclo[6.2.1.02.6]undec-2(6)-ene-4,9-dione (3f)

(82) To a solution of tert-butyl 7-(allyloxyamino)-3-[2-[tert-butyl(dimethyl)silyl]oxyethyl]-2-oxo-6,7-dihydro-4H-thiazolo[4,5-c]pyridine-5-carboxylate (3e) (4.29 g, 8.83 mmol) in DCM (88.3 mL) were added TEA (2.46 mL, 17.7 mmol) and diphosgene (1.58 mL, 11.48 mmol). After 5 min, the solution was washed with NaCl aqueous solution. The organic layer was dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The yellow oil obtained was stirred with 4M HCl solution in dioxane (88.3 mL) during 1 h. The mixture was concentrated in vacuo and to the crude in DCM (88.4 mL) was dropwise added TEA (2.25 mL, 17.67 mmol). The reaction mixture was stirred at rt for 30 min then diluted with DCM and washed with water. The organic layer was dried over Na.sub.2SO.sub.4 and concentrated in vacuo. The crude was purified by flash chromatography on silica gel (DCM/MeOH 100/0 to 90/10) to afford 10-allyloxy-5-(2-hydroxy-ethyl)-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2.6]undec-2(6)-ene-4,9-dione (3f) (1.72 g, 5.78 mmol, 66%).

(83) MS m/z ([M+H].sup.+) 298.

(84) .sup.1H NMR (300 MHz, CDCl.sub.3): (ppm) 3.27 (d, J=10.8 Hz, 1H), 3.52-3.78 (m, 7H), 4.18 (s, 1H), 4.33-4.47 (m, 2H), 5.28-5.38 (m, 2H), 5.90-6.04 (m, 1H).

Step 7: Preparation of Intermediate 10-allyloxy-5-(2-methylsulfonyloxyethyl)-3-thia-5,8,10-triaza-tricyclo[6.2.1.02.6]undec-2(6)-ene-4,9-dione (3g)

(85) To a solution of 10-allyloxy-5-(2-hydroxy-ethyl)-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2.6]undec-2(6)-ene-4,9-dione (3f) (316 mg, 1.06 mmol) in anhydrous pyridine (5.3 mL) at 0 C. was added MsCl (132 L, 1.70 mmol). The mixture was stirred for 2 h at 0 C. then concentrated in vacuo. The residue was diluted with DCM and washed with a 2N HCl solution. The organic layer was dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuo to provide 10-allyloxy-5-(2-methylsulfonyloxyethyl)-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2.6]undec-2(6)-ene-4,9-dione (3g) which was engaged in the next step without further purification.

(86) MS m/z ([M+H].sup.+) 376.

Step 8: Preparation of Intermediate 10-allyloxy-5-(2-(tert-butoxycarbonylamino)ethyl)-3-thia-5,8,10-triaza-tricyclo[6.2.1.02.6]undec-2(6)-ene-4,9-dione (3h)

(87) In a sealed flask, to a solution of 10-allyloxy-5-(2-methylsulfonyloxyethyl)-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2.6]undec-2(6)-ene-4,9-dione (3g) (1.06 mmol) in anhydrous DMF (5.30 mL) was added NaN.sub.3 (345 mg, 5.31 mmol). The reaction mixture was stirred 18 h at 65 C. before concentration in vacuo. The residue was dissolved in EtOAc and washed with NaCl aqueous solution. The organic layer was dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The yellow oil was dissolved in anhydrous THF (3.2 mL) and toluene (3.2 mL) and a solution of PMe.sub.3 1M in THF (1.6 mL) was added. The reaction mixture was stirred 1 h at rt then cooled to 0 C. and a solution of BocON (392 mg, 1.6 mmol) in anhydrous THF (2.2 mL) was slowly added. After 1 h at rt, the reaction mixture was concentrated in vacuo and purified by flash chromatography on silica gel (cyclohexane/EtOAc 100/0 to 0/100) to afford 10-allyloxy-5-(2-(tert-butoxycarbonylamino)ethyl)-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2.6]undec-2(6)-ene-4,9-dione (3h) (190 mg, 0.50 mmol, 45% over 2 steps) as a off white oil.

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

(89) .sup.1H NMR (300 MHz, CDCl.sub.3): (ppm) 1.41-1.42 (m, 9H), 3.27-3.36 (m, 2H), 3.47-3.55 (m, 1H), 3.65 (dd, J=2.8/10.9 Hz, 1H), 3.72-3.81 (m, 1H), 4.15 (dd, J=1.9/4.8 Hz, 2H), 4.34-4.48 (m, 2H), 4.37-4.90 (m, 1H), 5.28-5.38 (m, 2H), 5.91-6.05 (m, 1H), 7-13-7.24 (m, 1H).

Step 9: Preparation of Sodium [5-(2-(tert-butoxycarbonylamino)ethyl)-4,9-dioxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.02.6]undec-2(6)-en-10-yl] sulfate (Example 3)

(90) Using the procedure described in example 1 (step 7), 10-allyloxy-5-(2-(tert-butoxycarbonylamino)ethyl)-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2.6]undec-2(6)-ene-4,9-dione (3h) (190 mg, 0.48 mmol) is converted to sodium [5-(2-(tert-butoxycarbonylamino)ethyl)-4,9-dioxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2.6]undec-2(6)-en-10-yl] sulfate (example 3) (62.4 mg, 0.14 mmol, 28%) as a white solid.

(91) MS m/z ([MH].sup.) 435.

(92) .sup.1H NMR (400 MHz, D.sub.2O): (ppm) 1.36-1.38 (s, 9H), 3.24-3.40 (m, 2H), 3.44-3.53 (m, 1H), 3.62-3.67 (m, 2H), 3.82 (dd, J=2.99/11.6 Hz, 1H), 4.20-4.35 (m, 2H), 4.76 (m, in D.sub.2O peak, 1H).

Example 4

Synthesis of Sodium and 2,2,2-trifluoroacetate [5-(2-azaniumethyl)-4,9-dioxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.02.6]undec-2(6)-en-10-yl] sulfate

(93) ##STR00040##

Step 1: Preparation of Sodium and 2,2,2-trifluoroacetate [5-(2-azaniumethyl)-4,9-dioxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.02.6]undec-2(6)-en-10-yl] sulfate (Example 4)

(94) At 0 C., a solution of TFA (155 L) in DCM (105 L) was prepared and added to a solution of sodium [5-(2-(tert-butoxycarbonylamino)ethyl)-4,9-dioxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2.6]undec-2(6)-en-10-yl] sulfate (Example 3) (5.7 mg, 0.013 mmol) in DCM (155 L) at 0 C. The reaction mixture was stirred 30 min at this temperature then concentrated under nitrogen flux. The solid was dissolved in water (1 mL), filtered, frozen and lyophilized to afford sodium and 2,2,2-trifluoroacetate [5-(2-azaniumethyl)-4,9-dioxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2.6]undec-2(6)-en-10-yl] sulfate (Example 4) (3.2 mg, 0.007 mmol, 54%) as a white solid.

(95) MS m/z ([MH].sup.) 335.

(96) .sup.1H NMR (300 MHz, D.sub.2O): (ppm) 3.26 (t, J=6.0 Hz, 2H), 3.54 (d, J=11.5 Hz, 1H), 3.81 (dd, J=3.0/11.5 Hz, 1H), 3.92-3.98 (m, 2H), 4.25 (d, J=16.8 Hz, 1H), 4.38 (d, J=16.9 Hz, 1H), 4.70-4.86 (m, in D.sub.2O peak, 1H).

Example 5

Synthesis of Sodium [5-(2-hydroxyethyl)-4,9-dioxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.02.6]undec-2(6)-en-10-yl) sulfate

(97) ##STR00041##

Step 1: Preparation of Intermediate 10-Allyloxy-5-[2-[tert-butyl(dimethyl)silyl]oxyethyl]-3-thia-5,8,10-triaza-tricyclo[6.2.1.02.6]undec-2(6)-ene-4,9-dione (5a)

(98) To a solution of 10-allyloxy-5-(2-hydroxy-ethyl)-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2.6]undec-2(6)-ene-4,9-dione (3f) (150 mg, 0.50 mmol) in anhydrous DMF were added TBDMSCI (84 mg, 0.56 mmol) and imidazole (69 mg, 1.01 mmol). After 1 h at rt, the reaction mixture was diluted with EtOAc and washed with water. The organic layer was separated, dried over Na.sub.2SO.sub.4 and concentrated in vacuo. The crude was purified by flash chromatography on silica gel (cyclohexane/EtOAc 8/2 to 2/8) to afford 10-allyloxy-5-[2-[tert-butyl(dimethyl)silyl]oxyethyl]-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2.6]undec-2(6)-ene-4,9-dione (5a) (174 mg, 0.42 mmol, 83%).

(99) MS m/z ([M+H].sup.+) 412.

(100) .sup.1H NMR (400 MHz, CDCl.sub.3): (ppm) 0.02 (d, J=7.3 Hz, 6H), 0.86 (s, 9H), 3.16-3.23 (m, 1H), 3.37-3.46 (m, 1H), 3.63-3.86 (m, 4H), 4.16-4.21 (m, 3H), 4.35-4.50 (m, 2H), 5.28-5.39 (m, 2H), 5.93-6.06 (m, 1H).

Step 2: Preparation of Intermediate triphenyl-(propenyl)-phosphonium [5-[2-[tert-butyl(dimethyl)silyl]oxyethyl]-4,9-dioxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.02.6]undec-2(6)-en-10-yl] sulfate (5b)

(101) Using the procedure described in example 2 (step 7), 10-allyloxy-5-[2-[tert-butyl(dimethyl)silyl]oxyethyl]-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2.6]undec-2(6)-ene-4,9-dione (5a) (170 mg, 0.41 mmol) is converted to triphenyl-(propenyl)-phosphonium [5-[2-[tert-butyl(dimethyl)silyl]oxyethyl]-4,9-dioxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2.6]undec-2(6)-en-10-yl] sulfate (5b) (135 mg, 0.18 mmol 43%).

(102) MS m/z ([MH].sup.) 450.

Step 3: Preparation of Sodium [5-(2-hydroxyethyl)-4,9-dioxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.02.6]undec-2(6)-en-10-yl) sulfate (example 5)

(103) Triethylamine trihydrofluoride (29 L, 0.18 mmol) was added to a solution of triphenyl-(propenyl)-phosphonium [5-[2-[tert-butyl(dimethyl)silyl]oxyethyl]-4,9-dioxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2.6]undec-2(6)-en-10-yl] sulfate (5b) (135 mg, 0.18 mmol) in anhydrous ACN (1.46 mL) under inert atmosphere. The reaction mixture was vigorously stirred at 45 C. for 5 h until complete conversion of starting material. The reaction mixture was concentrated under nitrogen flow and directly applied on a Dowex sodium form column. The fractions containing the desired compound were combined and concentrated. The solid was diluted in a minimum of MeOH (300 L), filtered on Millipore to remove sodium fluoride salts and concentrated. The solid was diluted with water, freezed and lyophilized to afford compound sodium [5-(2-hydroxyethyl)-4,9-dioxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2.6]undec-2(6)-en-10-yl) sulfate (example 5) (16.6 mg, 0.05 mmol, 28%) as a white solid.

(104) MS m/z ([MH].sup.) 336.

(105) .sup.1H NMR (300 MHz, D.sub.2O): (ppm) 3.58 (dd, J=0.4/11.5 Hz, 1H), 3.74 (s, 4H), 3.85 (dd, J=3.0/11.5 Hz, 1H), 4.34 (d, J=18 Hz, 1H), 4.41 (d, J=18 Hz, 1H), 4.70-4.86 (m, in D.sub.2O peak, 1H).

Example 6

Synthesis of Sodium and 2,2,2-trifluoroacetate [5-(2-quanidiniumethyl)-4,9-dioxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.02.6]undec-2(6)-en-10-yl] sulfate

(106) ##STR00042##

Step 1: Preparation of Intermediate 10-allyloxy-5-(2-[[N,N1-bis(tert-butoxycarbonyl)carbamimidoyl]amino]ethyl)-3-thia-5,8,10-triaza-tricyclo[6.2.1.02.6]undec-2(6)-ene-4,9-dione (6a)

(107) To a solution of 10-allyloxy-5-(2-hydroxy-ethyl)-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2.6]undec-2(6)-ene-4,9-dione (3f) (130 mg, 0.44 mmol) in anhydrous THF (4.4 mL) under inert atmosphere was added 1,3-Bis(tert-butoxycarbonyl)guanidine (160 mg, 0.61 mmol), Ph.sub.3P (137 mg, 0.52 mmol), DIAD (106 mg, 0.52 mmol) and the mixture was stirred for 1 h30 at rt. The solution was filtered and concentrated under vacuum. The residue was purified by flash chromatography on silica gel (cyclohexane/EtOAc 80/20 to 20/80) then on preparative TLC (DCM/MeOH 96/4) to give 10-allyloxy-5-(2-[[N,N-bis(tert-butoxycarbonyl)carbamimidoyl]amino]ethyl)-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2.6]undec-2(6)-ene-4,9-dione (6a) (139.6 mg, 0.26 mmol, 59%) as a white solid.

(108) MS m/z ([M+H].sup.+) 539.

(109) MS m/z ([MH].sup.) 537.

(110) 1H NMR (300MHz, CDCl.sub.3): (ppm) 1.49 (s, 9H), 1.55 (s, 9H), 3.31 (d, J=10.6 Hz, 1H), 3.42-3.51 (m, 1H), 3.63 (dd, J=2.9/10.8 Hz, 1H), 3.87-4.06 (m, 2H), 4.13 (d, J=2.4 Hz, 1H), 4.15-4.27 (m, 2H), 4.34-4.48 (m, 3H), 5.27-5.38 (m, 2H), 5.94-6.03 (m,1H).

Step 2: Preparation of Intermediate Sodium [5-(2-[[N,N1-bis(tert-butoxycarbonyl)carbamimidoyl]amino]ethyl)-4,9-dioxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.02.6]undec-2(6)-en-10-yl] sulfate (6b)

(111) Using the procedure described in example 3 (step 9), 10-allyloxy-5-(2-[[N,N.sup.1-bis(tert-butoxycarbonyl)carbamimidoyl]amino]ethyl)-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2.6]undec-2(6)-ene-4,9-dione (6a) (139.6 mg, 0.26 mmol) is converted to sodium [5-(2-[[N,N.sup.1-bis(tert-butoxycarbonyl)carbamimidoyl]amino]ethyl)-4,9-dioxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2.6]undec-2(6)-en-10-yl] sulfate (6b) (30 mg, 0.05 mmol, 19% over 3 steps) as a white solid.

(112) MS m/z ([M+H].sup.+) 579.

(113) MS m/z ([MH].sup.) 577.

(114) 1H NMR (300 MHz, D.sub.2O): (ppm) 1.45 (s, 9H), 1.49 (s, 9H), 3.48 (d, J=11.4 Hz, 1H), 3.80 (d, J=2.9 Hz, 1H), 3.84 (d, J=3.0 Hz, 1H), 3.86-3.98 (m, 2H), 4.01-4.15 (m, 2H), 4.26-4.32 (m, 1H), 4.79 (m under D.sub.2O peak, 1H).

Step 3: Preparation of Sodium and 2,2,2-trifluoroacetate [5-(2-quanidiniumethyl)-4,9-dioxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.02.6]undec-2(6)-en-10-yl] sulfate (Example 6)

(115) At 0 C., a solution of TFA (636 L) in DCM (636 L) was prepared and added, drop by drop, to a solution of sodium [5-(2-[[N,N-bis(tert-butoxycarbonyl)carbamimidoyl]amino]ethyl)-4,9-dioxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2.6]undec-2(6)-en-10-yl] sulfate (6b) (17 mg, 0.028 mmol) in DCM (636 L) at 0 C. The mixture was stirred for 9 h at this temperature then concentrated under nitrogen flux. The residue was purified by chromatography C18 reverse phase to provide sodium and 2,2,2-trifluoroacetate [5-(2-guanidiniumethyl)-4,9-dioxo-3-thia-5,8,10-triaza-tricyclo[6.2.1.0.sup.2.6]undec-2(6)-en-10-yl] sulfate (Example 6) (2.3 mg, 4.4 mol, 16%) as a white solid.

(116) MS m/z ([M+H].sup.+) 379.

(117) MS m/z ([MH].sup.) 377.

(118) 1H NMR (300 MHz, DMSO): (ppm) 3.33 (m under D.sub.2O peak, 3H), 3.56 (dd, J=2.7/11.2 Hz, 1H), 3.66 (m, 2H), 4.11 (d, J=16.8 Hz, 1H), 4.22 (d, J=16.8 Hz, 1H), 4.56 (d, J=2.4 Hz, 1H), 7.00-7.34 (m, 3H), 7.55-7.59 (m, 1H).

(119) Compounds below could be obtained according to schemes 1-6.

(120) ##STR00043## ##STR00044##

Example 7

Biological Activity

Method 1: -lactamase Inhibitory Activity, Determination of IC50 (Table 1)

(121) 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).

(122) TABLE-US-00001 TABLE 1 IC.sub.50 (M) for -lactamase Inhibitory Activity IC.sub.50 -lactamase (M) (C) (A) AmpC AmpC (D) TEM-1 SHV-1 CTX-M-15 KPC-2 (P99) CMY-37 (PAE) OXA-1 OXA-11 OXA-15 OXA-48 Example 1 0.0033 0.012 0.00069 0.0033 0.048 0.042 0.19 0.063 0.040 0.25 0.00089 Example 2 0.00064 0.0013 0.0011 0.0013 0.016 0.030 0.22 0.038 0.0041 0.057 0.00067 Example 3 0.0058 0.020 0.00049 0.0034 0.0059 0.010 0.20 0.13 0.016 0.097 0.0017 Example 4 0.0046 0.021 0.0013 0.0074 0.14 0.22 1.2 0.46 0.082 0.30 0.0041 Example 5 0.0018 0.0060 0.00068 0.0017 0.024 0.062 0.37 0.31 0.031 0.19 0.00079 Example 6 0.0070 0.012 0.0054 0.0057 0.38 0.59 1.5 1.7 0.13 0.12 0.0016

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

(123) Compounds of the present invention were assessed against genotyped bacterial strains alone or in combination with the -lactam ceftazidime (CAZ). 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 (CLSI-M7-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 (Becton-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.

(124) TABLE-US-00002 TABLE 2 Bacterial species used in MIC determination Strains Resistance mechanism E. cloacae 260508 TEM-1, CTX-M-15 E. coli UFR61O TEM-1, KPC-2 K. pneumoniae BAA-1898 TEM-1, SHV-11, SHV-12, KPC-2 K. pneumoniae 160143 TEM-1, SHV-1, CTX-M-15, KPC-2, OXA-1 K. pneumoniae UFR68 TEM-1, SHV-11, CTX-M-15, KPC-3 E. cloacae P99 AmpC E. cloacae UFR85 TEM-1, CTX-M-15, derepressed AmpC E. cloacae UFR70 TEM-1, CTX-M-15, CMY-2, OXA-1, Porin loss K. pneumoniae UFR77 CMY-2 E. coli UFR74 SHV-1, DHA-1 E. coli UFR18 CTX-M-15, OXA-204 E. coli 131119 TEM-1, OXA-48 K. oxytoca UFR21 TEM-1, CTX-M-15, OXA-48 K. pneumoniae UFR24 TEM-1, SHV-2, SHV-11, OXA-1, OXA-48, OXA-47 K. pneumoniae 6299 TEM-1, SHV-11, OXA-163 E. coli RGN238 OXA-1 K. pneumoniae 200047 TEM-1, SHV-32, CTX-M-15, OXA-1 E. coli 190317 TEM-1, SHV-12, CTX-M-15, OXA-1 E. coli UFR32 TEM-1, VEB-1, OXA-10 E. cloacae UFR38 CTX-M-15, NDM-1 C. murliniae 210102 VIM-4 E. coli UFR52 TEM-1, SHV-12, IMP-8 P. aeruginosa CIP107051 TEM-24 P. aeruginosa CIP105250 OXA-15 P. aeruginosa UFR35 OXA-23 P. aeruginosa UFR90 derepressed AmpC, OprD P. aeruginosa UFR92 derepressed AmpC, OprD P. aeruginosa UFR93 derepressed AmpC, OprD, MexAB+, MexXY+ P. aeruginosa UFR47 VIM-1 P. aeruginosa UFR48 VIM-2 P. aeruginosa UFR59 IMP-29

(125) TABLE-US-00003 TABLE 3 MIC of compounds MIC compounds of the invention alone (g/mL) Exam- Exam- Exam- Exam- Exam- Exam- Strains ple 1 ple 2 ple 3 ple 4 ple 5 ple 6 260508 4 4 8 4 UFR61O 4 8 4 2 BAA-1898 16 32 >32 16 8 16 160143 8 16 16 8 UFR68 32 32 16 8 P99 8 16 >32 0.5 8 1 UFR85 8 8 2 2 UFR70 4 8 4 1 UFR77 8 4 16 4 UFR74 4 8 16 4 UFR18 4 4 0.5 2 131119 1 2 >32 2 1 UFR21 8 8 4 4 UFR24 8 16 16 4 6299 16 8 >32 32 8 >32 RGN238 2 1 >32 8 1 4 200047 4 4 8 2 190317 2 4 >32 1 1 1 UFR32 4 4 1 2 UFR38 8 8 0.5 2 210102 32 16 4 8 UFR52 8 8 2 0.5 CIP107051 >128 >32 >32 16 >32 8 CIP105250 >32 >32 >32 8 >32 8 UFR35 8 UFR90 8 UFR92 8 UFR93 16 UFR47 16 UFR48 8 UFR59 8

(126) TABLE-US-00004 TABLE 4 MIC of Ceftazidime/compound combinations combination of CAZ and compounds of the invention at 4 g/mL: MIC (g/mL) Strains CAZ Example 1 Example 2 Example 3 Example 4 Example 5 Example 6 260508 128 <0.25 <0.25 <0.25 <0.25 UFR61O 128 <0.25 <0.25 <0.25 <0.25 BAA-1898 256 64 >128 >128 0.125 4 0.125 160143 128 0.5 8 0.25 <0.25 UFR68 >128 64 32 <0.25 0.25 P99 128 0.5 4 128 <0.25 0.25 <0.25 UFR85 128 <0.25 16 <0.25 <0.25 UFR70 >128 <0.25 1 <0.25 <0.25 UFR77 64 0.25 0.25 0.25 <0.25 UFR74 64 <0.25 1 0.25 <0.25 UFR18 >128 <0.25 0.5 <0.25 <0.25 131119 0.5 <0.25 <0.25 <0.25 <0.25 UFR21 128 2 16 <0.25 <0.25 UFR24 >128 4 4 0.25 <0.25 6299 256 4 4 128 0.125 0.125 0.125 RGN238 0.5 <0.25 <0.25 <0.25 <0.25 200047 128 <0.25 <0.25 <0.25 <0.25 190317 128 0.125 <0.25 32 <0.25 <0.25 <0.25 UFR32 >128 <0.25 <0.25 <0.25 <0.25 UFR38 >128 >128 >128 <0.25 <0.25 210102 >128 >128 >128 <0.25 64 UFR52 >128 0.5 >128 <0.25 <0.25 CIP107051 256 4 16 64 4 4 8 UFR35 2 4 2 UFR90 64 64 1 UFR92 32 32 0.25