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NOVEL HETEROCYCLIC COMPOUNDS AND THEIR USE IN PREVENTING OR TREATING BACTERIAL INFECTIONS

20180086761 · 2018-03-29

    Inventors

    Cpc classification

    International classification

    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.-22. (canceled)

    23. A compound of formula (I) ##STR00038## wherein: 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 saturated or partially unsaturated or totally unsaturated or aromatic and optionally substituted by one or more T.sup.1, CN, (CH.sub.2).sub.mOQ.sup.1, (CH.sub.2).sub.mOC(O)Q.sup.1, C(O)OQ.sup.1, (CH.sub.2).sub.mOC(O)Q.sup.1, (CH.sub.2).sub.mOC(O)NQ.sup.1Q.sup.2, C(O)NHQ.sup.1, C(O)NHOQ.sup.1, (CH.sub.2).sub.mNHS(O).sub.2NQ.sup.1Q.sup.2, C(O)NHNHQ.sup.1, C(O)ONHQ.sup.1, (CH.sub.2).sub.mNHC(O)Q.sup.1, (CH.sub.2).sub.mHS(O).sub.2Q.sup.1, (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 a 4- to 10-member heterocycle that is saturated or partially unsaturated or totally unsaturated or aromatic and optionally substituted by one or more T.sup.2; R.sup.3 is SO.sub.3H, CFHCOOH, or CF.sub.2COOH; T.sup.1, identical or different, is independently selected from the group consisting of F, C.sub.1-C.sub.3 alkyl that is optionally substituted by one or more T.sup.3, C.sub.1-C.sub.3 fluoroalkyl that is optionally substituted by one or more T.sup.3, OC.sub.1-C.sub.3 fluoroalkyl that is optionally substituted by one or more T.sup.3, (CH.sub.2).sub.n-(4- or 5- or 6-member heterocycle comprising at least one nitrogen atom that is saturated or partially unsaturated or totally unsaturated or aromatic) that is optionally substituted by one or more T.sup.3, (CH.sub.2).sub.nOQ.sup.1, (CH.sub.2).sub.nC(O)ONHQ.sup.1, (CH.sub.2).sub.nCN, (CH.sub.2).sub.nOC(O)Q.sup.1, (CH.sub.2).sub.nC(O)OQ.sup.1, (CH.sub.2).sub.nNHS(O).sub.2NQ.sup.1Q.sup.2, (CH.sub.2).sub.nOC(O)OQ.sup.1, (CH.sub.2).sub.nOC(O)NHQ.sup.1, (CH.sub.2).sub.nC(O)NHQ.sup.1, (CH.sub.2).sub.nC(O)NHOQ.sup.1, (CH.sub.2).sub.nC(O)NHNHQ.sup.1, (CH.sub.2).sub.nNHC(O)Q.sup.1, (CH.sub.2).sub.nNHS(O).sub.2Q.sup.1, (CH.sub.2).sub.nNHC(O)OQ.sup.1, (CH.sub.2).sub.nNHC(O)NQ.sup.1Q.sup.2, (CH.sub.2).sub.nNHQ.sup.1, (CH.sub.2).sub.nNHC(NHQ.sup.3)=NQ.sup.4, (CH.sub.2).sub.nNHCHNQ.sup.3, and (CH.sub.2).sub.nC(NHQ.sup.3)=NQ.sup.4; Q.sup.1 and Q.sup.2 is one of the following: Q.sup.1 and Q.sup.2, identical or different, are independently selected from the group consisting of H, C.sub.1-C.sub.3 alkyl that is optionally substituted by one or more T.sup.3, (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.rC(NHQ.sup.3)=NQ.sup.4, (CH.sub.2).sub.qOQ.sup.3, (CH.sub.2).sub.rCONHQ.sup.3, (CH.sub.2).sub.n-(4- or 5- or 6-member heterocycle comprising at least one nitrogen atom that is saturated or partially unsaturated or totally unsaturated or aromatic) that is optionally substituted by one or more T.sup.3; or Q.sup.1 and Q.sup.2 and the nitrogen atom to which they are bonded form together 4- or 5- or 6-member heterocycle that is saturated or partially unsaturated and optionally substituted by one or more T.sup.3; Q.sup.3 and Q.sup.4, identical or different, are independently selected from the group consisting of H and C.sub.1-C.sub.3 alkyl; T.sup.2, identical or different, is independently selected from the group consisting of F, C.sub.1-C.sub.3 alkyl that is optionally substituted by one or more T.sup.3, C.sub.1-C.sub.3 fluoroalkyl that is optionally substituted by one or more T.sup.3, OC.sub.1-C.sub.3 fluoroalkyl that is optionally substituted by one or more T.sup.3, (X).sub.p(CH.sub.2).sub.nC.sub.3-C.sub.6 cycloalkyl that is optionally substituted by one or more T.sup.3, (X).sub.p(CH.sub.2).sub.nC.sub.3-C.sub.6 cyclofluoroalkyl, (X).sub.p(CH.sub.2).sub.n-(4- or 5- or 6-member heterocycle that is saturated or partially unsaturated or totally unsaturated or aromatic) that is optionally substituted by one or more T.sup.3, (X).sub.p(CH.sub.2).sub.tOQ.sup.5, (X).sub.p(CH.sub.2).sub.uCN, (X)(CH.sub.2).sub.tOC(O)Q.sup.5, (X).sub.p(CH.sub.2).sub.uC(O)OQ.sup.5, (X).sub.p(CH.sub.2).sub.tOC(O)OQ.sup.5, (X).sub.p(CH.sub.2).sub.tOC(O)NQ.sup.5Q.sup.6, (X).sub.p(CH.sub.2).sub.uC(O)NQ.sup.5Q.sup.6, (X).sub.p(CH.sub.2).sub.uC(O)ONQ.sup.5Q.sup.6, (X).sub.p(CH.sub.2).sub.uC(O)NQ.sup.5OQ.sup.6, (X).sub.p(CH.sub.2).sub.uC(O)NQ.sup.5-NQ.sup.5Q.sup.6, (X).sub.p(CH.sub.2).sub.t-NQ.sup.5C(O)Q.sup.6, (X).sub.p(CH.sub.2).sub.t-NQ.sup.5S(O).sub.2NQ.sup.5Q.sup.6, (X).sub.p(CH.sub.2).sub.t-NQ.sup.5S(O).sub.2Q.sup.6, (X).sub.p(CH.sub.2).sub.t-NQ.sup.5C(O)OQ.sup.6, (X).sub.p(CH.sub.2).sub.tNQ.sup.5C(O)NQ.sup.5Q.sup.6, (X).sub.p(CH.sub.2).sub.t-NQ.sup.5Q.sup.6, (X).sub.p(CH.sub.2).sub.tNHC(NHQ.sup.3)=NQ.sup.4, (X).sub.p(CH.sub.2).sub.tNHCHNQ.sup.3, (X).sub.p(CH.sub.2).sub.uC(NHQ.sup.3)=NQ.sup.4; Q.sup.5 and Q.sup.6 is one of the following: Q.sup.5 and Q.sup.6, identical or different, are independently selected from the group consisting of H, C.sub.1-C.sub.3 alkyl that is optionally substituted by one or more T.sup.3, (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.rC(NHQ.sup.3)=NQ.sup.4, (CH.sub.2).sub.qOQ.sup.3, (CH.sub.2).sub.rCONHQ.sup.3, and (CH.sub.2).sub.n-(4- or 5- or 6-member heterocycle comprising at least one nitrogen atom that is saturated or partially saturated totally unsaturated or aromatic) that is optionally substituted by one or more T.sup.3; or Q.sup.5 and Q.sup.6 and the nitrogen atom to which they are bonded form together a saturated or partially unsaturated 4- to 6-member heterocycle that is optionally substituted by one or more T.sup.3; T.sup.3, identical or different, is independently selected from the group consisting of OH, NH.sub.2, and CONH.sub.2; m, identical or different, is independently selected from the group consisting of 1 and 2; n, identical or different, is independently selected from the group consisting of 0, 1, 2, and 3; t, identical or different, is independently selected from the group consisting of 0, 1, 2, and 3 if p is 0, but if p is 1 then t, identical or different, is independently selected from the group consisting of 2 and 3; u, identical or different, is independently selected from the group consisting of 0, 1, 2, and 3 if p is 0, but if p is 1 then u, identical or different is independently selected from the group consisting of 1, 2, and 3; q, identical or different, is independently selected from the group consisting of 2 and 3; r, identical or different, is independently selected from the group consisting of 1, 2, and 3; p, identical or different, is independently selected from the group consisting of 0 and 1; X, identical or different, is independently selected from the group consisting of O, S, S(O), S(O).sub.2 and N(Q.sup.3); wherein any carbon atom present within any of the foregoing alkyls, cycloalkyls, fluoralkyls, cyclofluoroalkyls, heterocycles may be oxidized to form a C(O) group; wherein any sulphur atom present within any of the foregoing heterocycles may be oxidized to form a S(O) group or a S(O).sub.2 group; wherein any nitrogen atom present within a heterocycle or a tertiary amine group may be further quaternized by a methyl group; or a pharmaceutically acceptable salt, a corresponding zwitterion, an optical isomer, a racemate, a diastereoisomer, an enantiomers, or a tautomer of formula (I).

    24. The compound according to claim 23, wherein R1 is one of the following: 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 saturated or partially unsaturated, or totally unsaturated or aromatic and is optionally substituted by one or more T.sup.1, CN, (CH.sub.2).sub.mOQ.sup.1, (CH.sub.2).sub.mOC(O)Q.sup.1, C(O)OQ.sup.1, (CH.sub.2).sub.mOC(O)OQ.sup.1, (CH.sub.2).sub.mOC(O)NQ.sup.1Q.sup.2, C(O)NHQ.sup.1, (CH.sub.2).sub.mNHS(O).sub.2NQ.sup.1Q.sup.2, C(O)NHOQ.sup.1, C(O)NHNHQ.sup.1, C(O)ONHQ.sup.1, (CH.sub.2).sub.mNHC(O)Q.sup.1, (CH.sub.2).sub.mNHS(O).sub.2Q.sup.1, (CH.sub.2).sub.mNHC(O)OQ.sup.1, (CH.sub.2).sub.mNHC(O)NQ.sup.1Q.sup.2; or R.sup.1 is selected from the group consisting of (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.

    25. The compound according to claim 23, wherein R1 is one of the following: 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 saturated or partially unsaturated or totally unsaturated or aromatic 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 and (CH.sub.2).sub.mOQ.sup.1; or R.sup.1 is selected from the group consisting of (CH.sub.2).sub.mNHQ.sup.3, (CH.sub.2).sub.mNHC(NHQ.sup.3)=NQ.sup.4, wherein m is 1, and wherein Q.sup.3 and Q.sup.4 are H.

    26. The compound according to claim 23, wherein R.sup.1 is selected from the group consisting of CN, C(O)OQ.sup.1, C(O)NHQ.sup.1, C(O)NHOQ.sup.1, and C(O)NHNHQ.sup.1.

    27. The compound according to claim 23, wherein 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.

    28. The compound according to claim 23, wherein 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.

    29. The compound according to claim 23, wherein R.sup.1 is selected from the group consisting of (CH.sub.2).sub.mNHQ.sup.3, (CH.sub.2).sub.mNHC(NHQ.sup.3)=NQ.sup.4, CN, C(O)NHQ.sup.1, C(O)NHOQ.sup.1, C(O)NHNHQ.sup.1, (CH.sub.2).sub.mOQ.sup.1, (CH.sub.2).sub.mNHC(O)Q.sup.1, (CH.sub.2).sub.mNHC(O)NQ.sup.1Q.sup.2, (CH.sub.2).sub.mNHC(O)OQ.sup.1, and 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 heteroatom is selected from the group consisting of N, O, S, S(O) and S(O).sub.2; and wherein Q.sup.3 and Q.sup.4 are H.

    30. The compound according to claim 23, wherein R.sup.2 is selected from the group consisting of a 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 optional substituted by one or more T.sup.2 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.

    31. A compound according to claim 23, wherein: R.sup.1 is one of the following: R.sup.1 is (CH.sub.2).sub.mNHQ.sup.3 or (CH.sub.2).sub.mNHC(NHQ.sup.3)=NQ.sup.4, wherein m is 1; or 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; 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; 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 and is monocyclic; R.sup.1 is selected from the group consisting of (CH.sub.2).sub.mNHQ.sup.3, (CH.sub.2).sub.mNHC(NHQ.sup.3)=NQ.sup.4, (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, (CH.sub.2).sub.mNHC(O)NQ.sup.1Q.sup.2, 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; and R.sup.2 is monocyclic or bicyclic; Q.sup.1 and Q.sup.2, identical or different, are independently selected from the group consisting of H, methyl, CH.sub.2CH.sub.2NH.sub.2, CH.sub.2CH.sub.2NHCNH.sub.2NH, CH.sub.2CH.sub.2NHCHNH, CH.sub.2C(NH.sub.2)NH, CH.sub.2CH.sub.2OH, CH.sub.2CONH.sub.2, and (CH.sub.2).sub.n-(4- or 5- or 6-member heterocycle comprising at least one nitrogen atom that saturated or partially unsaturated or totally unsaturated or aromatic and optionally substituted by one or more T.sup.3); and Q.sup.3 and Q.sup.4 are H.

    32. The compound according to claim 23, wherein: R.sup.1 is one of the following: R.sup.1 is selected from the group consisting of (CH.sub.2).sub.mNHQ.sup.3 and (CH.sub.2).sub.mNHC(NHQ.sup.3)=NQ.sup.4, wherein m is 1; 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; R.sup.1 is selected from the group consisting of (CH.sub.2).sub.mOQ.sup.1 and (CH.sub.2).sub.mNHC(O)OQ.sup.1; 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 and is monocyclic; or R.sup.1 is selected from the group consisting of (CH.sub.2).sub.mNHQ.sup.3, (CH.sub.2).sub.mNHC(NHQ.sup.3)=NQ.sup.4, (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, (CH.sub.2).sub.mNHC(O)NQ.sup.1Q.sup.2, 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; R.sup.2 is a monocyclic or bicyclic 4- to 10-member heterocycle that is saturated or partially unsaturated or totally unsaturated or aromatic and optionally substituted by one or more T.sup.2; R.sup.3 is SO.sub.3H or CF.sub.2COOH; Q.sup.1 and Q.sup.2, identical or different, independently selected from the group consisting of H, methyl, CH.sub.2CH.sub.2NH.sub.2, CH.sub.2CH.sub.2NHCNH.sub.2NH, CH.sub.2CH.sub.2NHCHNH, CH.sub.2C(NH.sub.2)NH, CH.sub.2CH.sub.2OH, CH.sub.2CONH.sub.2, a (CH.sub.2).sub.n-(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.3; and Q.sup.3 and Q.sup.4 are H.

    33. The compound according to claim 23 of formula (I*) ##STR00039##

    34. The compound according to claim 23 selected from the group consisting of ##STR00040## wherein: X is selected from the group consisting of halogen, B(OR).sub.2, OTf, and SnR.sub.3, wherein R is alkyl or the OR are linked together with the B to form a cycle; and PG.sub.1 and PG.sub.2, which are different, are protective groups.

    35. The compound according to claim 23 selected from the group consisting of ##STR00041## wherein: X is selected from the group consisting of halogen, B(OR).sub.2, OTf, and SnR.sub.3, wherein R is alkyl or the OR are linked together with the B to form a cycle comprising 5 members; and PG.sub.1 and PG.sub.2, which are different, are protective groups selected from the group consisting of allyl, benzyl, tertbutyldimethylsilyl (TBDMS), and tert-butoxycarbonyl (Boc).

    36. A pharmaceutical composition comprising at least one compound according to claim 23 with a pharmaceutically acceptable excipient.

    37. The pharmaceutical composition according to claim 36, further comprising an antibacterial agent selected from group consisting of aminoglycosides, beta-lactams, glycylcyclines, tetracyclines, quinolones, fluoroquinolones, glycopeptides, lipopeptides, macrolides, ketolides, lincosamides, streptogramins, oxazolidinones, polymyxins, and combinations thereof.

    38. The pharmaceutical composition according to claim 13, wherein the antibacterial agent is selected from the group consisting of penicillin, cephalosporins, penems, carbapenems, monobactam, and combinations thereof.

    39. A pharmaceutical composition comprising the compound according to claim 23 and ceftazidime.

    40. A kit comprising: a pharmaceutical composition according to claim 34; and at least another composition comprising one or more antibacterial agent(s).

    41. A kit according comprising: a pharmaceutical composition comprising at least a compound according to claim 23; and a pharmaceutical composition comprising ceftazidime.

    42. A method for the treatment or prevention of bacterial infections in a patient, the method comprising the administration of a therapeutically effective amount of a compound according to claim 23.

    43. The method according to claim 42, wherein the bacterial infection is caused by bacteria producing one or more beta-lactamases.

    44. The method according to claim 42, wherein the bacterial infection is caused by gram-negative bacteria.

    Description

    EXAMPLES

    [0247] 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.

    [0248] The first part represents the preparation of the compounds (intermediates and final compounds) whereas the second part describes the evaluation of antibacterial activity of compounds according to the invention.

    Example 1: synthesis of sodium and 2,2,2-trifluoroacetate [trans-2-(azaniumylmethyl)-4-oxazol-5-yl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl] sulfate

    [0249] ##STR00028## ##STR00029##

    Step 1: Preparation of Intermediate tert-butyl trans-3-[allyloxy-(2-nitrophenyl)sulfonyl-amino]-6-[[tert-butyl(dimethyl)silyl]oxymethyl]-4-iodo-3,6-dihydro-2H-pyridine-1-carboxylate (2)

    [0250] To a solution of tert-butyl cis-6-[[tert-butyl(dimethyl)silyl]oxymethyl]-3-hydroxy-4-iodo-3,6-dihydro-2H-pyridine-1-carboxylate (1, prepared according to WO 2013/150296) (12.05 g, 25.67 mmol) in toluene (170 mL) at rt was added triphenylphosphine (8.08 g, 30.80 mmol), N-(allyloxy)-2-nitrobenzenesulfonamide (6.63 g, 25.67 mmol) and DIAD (6.06 mL, 30.80 mmol). The reaction mixture was stirred at rt overnight and concentrated in vacuo. The crude was purified by flash chromatography on silica gel (cyclohexane/EtOAc 100/0 to 85/15) to give tert-butyl trans-3-[allyloxy-(2-nitrophenyl)sulfonyl-amino]-6-[[tert-butyl(dimethyl)silyl]oxymethyl]-4-iodo-3,6-dihydro-2H-pyridine-1-carboxylate (2) (17.0 g, 23.95 mmol, 93%).

    [0251] .sup.1H NMR (300 MHz, CDCl.sub.3): (ppm) 0.03 (s, 6H), 0.88 (s, 9H), 1.35 (s, 9H), 3.16-3.75 (m, 3H), 3.93-4.78 (m, 5H), 5.12-5.38 (m, 2H), 5.68-5.89 (m, 1H), 6.73 (d, J=4.1 Hz, 1H), 7.54-7.66 (m, 1H), 7.69-7.84 (m, 2H), 8.06-8.19 (m, 1H).

    Step 2: Preparation of Intermediate trans-N-allyloxy-6-[[tert-butyl(dimethyl)silyl]oxymethyl]-4-iodo-1,2,3,6-tetrahydropyridin-3-amine (3)

    [0252] To a solution of tert-butyl trans-3-[allyloxy-(2-nitrophenyl)sulfonyl-amino]-6-[[tert-butyl(dimethyl)silyl]oxymethyl]-4-iodo-3,6-dihydro-2H-pyridine-1-carboxylate (2) (17.0 g, 23.95 mmol) in DCM (177 mL) was added ZnBr.sub.2 (16.2 g, 71.86 mmol). The reaction mixture was stirred at rt overnight then diluted with DCM and successively washed with saturated and brine. The organic layer was dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The crude was diluted with ACN (177 mL). K.sub.2CO.sub.3 (16.6 g, 119.77 mmol) was added, followed by thiophenol (12.3 mL, 119.77 mmol). The reaction mixture was stirred at rt for 1 h and concentrated in vacuo. DCM was added and the resulting solids were removed by filtration. The filtrate was concentrated in vacuo. The crude was purified by flash chromatography on silica gel (DCM/MeOH 100/0 to 90/10) to give trans-N-allyloxy-6-[[tert-butyl(dimethyl)silyl]oxymethyl]-4-iodo-1,2,3,6-tetrahydropyridin-3-amine (3) (7.99 g, 18.83 mmol, 78%).

    [0253] MS m/z ([M+H].sup.+) 425.

    [0254] .sup.1H NMR (300 MHz, CDCl.sub.3): (ppm) 0.06 (s, 6H), 0.89 (s, 9H), 1.82 (bs, 1H), 3.14 (dd, J=12.6, 5.1 Hz, 1H), 3.21 (dd, J=12.6, 3.9 Hz, 1H), 3.37-3.45 (m, 2H), 3.53-3.60 (m, 2H), 4.22 (dq, J=6.0, 1.2 Hz, 2H), 5.18-5.25 (m, 1H), 5.25-5.35 (m, 1H), 5.88-5.35 (m, 2H), 6.53-6.56 (m, 1H).

    Step 3: Preparation of Intermediate trans-6-allyloxy-2-[[tert-butyl(dimethyl)silyl]oxymethyl]-4-iodo-1,6-diazabicyclo[3.2.1]oct-3-en-7-one (4)

    [0255] To a solution of trans-N-allyloxy-6-[[tert-butyl(dimethyl)silyl]oxymethyl]-4-iodo-1,2,3,6-tetrahydropyridin-3-amine (3) (7.99 g, 18.83 mmol) in anhydrous ACN (980 mL) at 0 C. under inert atmosphere was added TEA (10.56 mL, 75.31 mmol). A solution of diphosgene (1.14 mL, 9.41 mmol) in anhydrous ACN (20 mL) was dropwise added over 5 h. Once the addition finished, the reaction mixture is allowed to reach rt and stirred for 3 days. H.sub.2O was added and the mixture was extracted with EtOAc. The organic layer was dried over Na.sub.2SO.sub.4, filtered and evaporated in vacuo. The crude was purified by flash chromatography on silica gel (cyclohexane/EtOAc 100/0 to 80/20) to give trans-6-allyloxy-2-[[tert-butyl(dimethyl)silyl]oxymethyl]-4-iodo-1,6-diazabicyclo[3.2.1]oct-3-en-7-one (4) (7.25 g, 16.10 mmol, 85%).

    [0256] MS m/z ([M+H].sup.+) 451.

    [0257] .sup.1H NMR (300 MHz, CDCl.sub.3): (ppm) 0.06 (s, 6H), 0.88 (s, 9H), 3.19 (dd, J=11.1, 3.0 Hz, 1H), 3.57 (d, J=11.1 Hz, 1H), 3.80-3.90 (m, 3H), 4.05-4.08 (m, 1H), 4.35-4.53 (m, 2H), 5.28-5.34 (m, 1H), 5.34-5.43 (m, 1H), 5.97-6.12 (m, 1H), 6.37-6.41 (m, 1H).

    Step 4: Preparation of Intermediate trans-6-allyloxy-2-[[tert-butyl(dimethyl)silyl]oxymethyl]-4-oxazol-5-yl-1,6-diazabicyclo[3.2.1]oct-3-en-7-one (5)

    [0258] In a sealed flask, a mixture of trans-6-allyloxy-2-[[tert-butyl(dimethyl)silyl]oxymethyl]-4-iodo-1,6-diazabicyclo[3.2.1]oct-3-en-7-one (4) (5.20 g, 11.55 mmol), 5-(4,4,5,5-tetramethyl-1,2,3-dioxaborolan-2-yl)oxazole (2.70 g, 13.86 mmol) and CsCO.sub.3 (7.52 g, 23.09 mmol) in anhydrous THF (100 mL) was degassed under argon for 5 min and Pd(PPh.sub.3).sub.4 (400 mg, 0.35 mmol) was added. The mixture was heated at 60 C. overnight. H.sub.2O was added and the mixture was extracted with EtOAc. The organic layer was dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The crude was purified by flash chromatography on silica gel (cyclohexane/EtOAc 100/0 to 70/30) to give trans-6-allyloxy-2-[[tert-butyl(dimethyl)silyl]oxymethyl]-4-oxazol-5-yl-1,6-diazabicyclo[3.2.1]oct-3-en-7-one (5) (3.64 g, 9.30 mmol, 80%).

    [0259] MS m/z ([M+H].sup.+) 392.

    [0260] .sup.1H NMR (300 MHz, CDCl.sub.3): (ppm) 0.07 (s, 6H), 0.88 (s, 9H), 3.37 (dd, J=11.0, 3.0 Hz, 1H), 3.54 (d, J=11.0 Hz, 1H), 3.87-4.06 (m, 3H), 4.11-4.14 (m, 1H), 4.33-4.50 (m, 2H), 5.27-5.40 (m, 2H), 5.92-6.08 (m, 1H), 6.15 (d, J=3.0 Hz, 1H), 7.03 (s, 1H), 7.83 (s, 1H).

    Step 5: Preparation of Intermediate trans-6-allyloxy-2-(hydroxymethyl)-4-oxazol-5-yl-1,6-diazabicyclo[3.2.1]oct-3-en-7-one (6)

    [0261] To a solution of trans-6-allyloxy-2-[[tert-butyl(dimethyl)silyl]oxymethyl]-4-oxazol-5-yl-1,6-diazabicyclo[3.2.1]oct-3-en-7-one (5) (3.64 g, 9.30 mmol) in THF (45 mL) at 0 C. was added tetrabutylammonium fluoride (1M in tetrahydrofuran) (13.9 mL, 13.94 mmol). The reaction mixture was stirred at 0 C. for 1 h and concentrated in vacuo. The crude was purified by flash chromatography on silica gel (EtOAc 100%) to give trans-6-allyloxy-2-(hydroxymethyl)-4-oxazol-5-yl-1,6-diazabicyclo[3.2.1]oct-3-en-7-one (6) (1.53 g, 5.52 mmol, 57%).

    [0262] MS m/z ([M+H].sup.+) 278.

    [0263] .sup.1H NMR (400 MHz, CDCl.sub.3): (ppm) 3.32 (d, J=11.2 Hz, 1H), 3.40 (dd, J=11.2, 2.9 Hz, 1H), 3.69-3.87 (m, 2H), 4.12-4.19 (m, 2H), 4.36-4.50 (m, 2H), 5.28-5.39 (m, 3H), 5.94-6.06 (m, 2H), 7.06 (s, 1H), 7.83 (s, 1H).

    Step 6: Preparation of Intermediate tert-butyl N-[[trans-6-allyloxy-4-oxazol-5-yl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-2-yl]methyl]carbamate (7)

    [0264] A solution of (trans-6-allyloxy-2-(hydroxymethyl)-4-oxazol-5-yl-1,6-diazabicyclo[3.2.1]oct-3-en-7-one (6) (1.53 g, 5.52 mmol) in pyridine (17 mL) was cooled to 0 C. Methanesulfonyl chloride (0.67 mL, 8.61 mmol) was added and the reaction mixture was stirred at the same temperature for 2 h. After concentrating in vacuo, the crude was dissolved in DCM and successively washed with a solution of 1N HCl and brine. The organic layer was dried over Na.sub.2SO.sub.4, filtered and evaporated in vacuo. The crude was dissolved in DMF (29 mL) and NaN.sub.3 (1.79 g, 27.59 mmol) was added. The reaction mixture was heated at 65 C. overnight and concentrated in vacuo. H.sub.2O was added to the crude, which was extracted with EtOAc. The organic layer was washed with brine, dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The crude was dissolved in a mixture of THF and toluene (16.7 mL/16.7 mL) and trimethylphosphine (1M in tetrahydrofuran) (8.28 mL, 8.28 mmol) was added at 0 C. After 1 h stirring at rt, the mixture was cooled to 0 C. and a solution of 2-(Boc-oxyimino)-2-phenylacetonitrile (2.04 g, 8.28 mmol) in THF (11 mL) was dropwise added. The mixture was stirred at rt for 1 h and concentrated in vacuo. The crude was purified by flash chromatography on silica gel (cyclohexane/EtOAc 95/5 to 0/100) to give tert-butyl N-[[trans-6-allyloxy-4-oxazol-5-yl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-2-yl]methyl]carbamate (7) (440 mg, 1.17 mmol, 21%).

    [0265] .sup.1H NMR (300 MHz, CDCl.sub.3): (ppm) 1.45 (s, 9H), 3.11-3.30 (m, 1H), 3.37 (dd, J=11.3, 2.9 Hz, 1H), 3.53-3.67 (m, 1H), 3.98-4.07 (m, 1H), 4.15 (d, J=2.9 Hz, 1H), 4.33-4.50 (m, 2H), 4.99-5.12 (m, 1H), 5.28-5.41 (m, 2H), 5.92-6.07 (m, 2H), 7.05 (s, 1H), 7.83 (s, 1H).

    [0266] MS m/z ([M+H].sup.+) 377.

    Step 7: Preparation of Intermediate triphenyl-[(E)-prop-1-enyl]phosphonium [trans-2-[(tert-butoxycarbonylamino)methyl]-4-oxazol-5-yl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl]sulfate (8)

    [0267] To a solution of tert-butyl N-[[trans-6-allyloxy-4-oxazol-5-yl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-2-yl]methyl]carbamate (7) (440 mg, 1.17 mmol) and glacial acetic acid (134 L, 2.34 mmol) in anhydrous DCM (13 mL) was added in one portion Pd(PPh.sub.3).sub.4 (675 mg, 0.58 mmol). After stirring for 2 h, a solution of sulfur trioxide pyridine complex (753 mg, 4.73 mmol) in dry pyridine (15 mL) was added and the resulting mixture was stirred overnight. The reaction mixture was concentrated in vacuo, diluted with DCM and filtered. The filtrate was concentrated in vacuo. The crude was purified by flash chromatography on silica gel (DCM/acetone 97/3 to 20/80) to give triphenyl-[(E)-prop-1-enyl]phosphonium [trans-2-[(tert-butoxycarbonylamino)methyl]-4-oxazol-5-yl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl] sulfate (8) (560 mg, 0.78 mmol, 67%).

    [0268] .sup.1H NMR (300 MHz, CDCl.sub.3): (ppm) 1.45 (s, 9H), 2.23-2.28 (m, 3H), 3.07-3.31 (m, 2H), 3.46-3.67 (m, 2H), 3.91-4.01 (m, 1H), 4.77 (bs, 1H), 5.10-5.27 (m, 1H), 5.85 (bs, 1H), 6.52-6.70 (m, 1H), 7.11-7.24 (m, 1H), 7.60-7.82 (m, 1H).

    Step 8: Preparation of sodium and 2,2,2-trifluoroacetate [trans-2-(azaniumylmethyl)-4-oxazol-5-yl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl] sulfate (Example 1)

    [0269] A solution of triphenyl-[(E)-prop-1-enyl]phosphonium [trans-2-[(tertbutoxycarbonylamino) methyl]-4-oxazol-5-yl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl] sulfate (8) (560 mg, 0.78 mmol) dissolved in a mixture of H.sub.20/THF 7/3 (1 mL) 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 H.sub.2O). The fractions containing the desired compound were combined and concentrated in vacuo. The product was dissolved in ACN and the remaining precipitate was filtered off. The filtrate was concentrated in vacuo. The crude was dissolved in DCM (28 mL), cooled to 0 C., and trifluoroacetic acid (18.5 mL) was dropwise added. After 1 h stirring at the same temperature, the reaction mixture was concentrated in vacuo, dissolved in a minimum of H.sub.2O, freezed and lyophilized to afford sodium and 2,2,2-trifluoroacetate [trans-2-(azaniumylmethyl)-4-oxazol-5-yl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl] sulfate (example 1) (350 mg, 0.77 mmol, 99%).

    [0270] .sup.1H NMR (300 MHz, DMSO-d.sub.6): (ppm) 3.10-3.26 (m, 1H), 3.29 (dd, J=11.7, 2.9 Hz, 1H), 3.46 (d, J=11.7 Hz, 1H), 4.02-4.11 (m, 1H), 4.59 (d, J=2.2 Hz, 1H), 6.00 (d, J=3.3 Hz, 1H), 7.30 (s, 1H), 8.09 (bs, 3H), 8.42 (s, 1H).

    Example 2: synthesis of sodium and 2,2,2-trifluoroacetate [(2S,5R)-2-(azaniumylmethyl)-7-oxo-3-thiazol-2-yl-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl] sulfate

    [0271] ##STR00030## ##STR00031##

    Step 1: Preparation of Intermediate tert-butyl N-[(1R)-1-(hydroxymethyl)-2-[methoxy(methyl)amino]-2-oxo-ethyl]carbamate (10)

    [0272] To a solution of Boc-D-Ser-OH (9) (5 g, 24.37 mmol) in anhydrous DCM (100 mL) at 15 C. were added N,O-dimethylhydroxylamine hydrochloride (2.54 g, 26.07 mmol) and N-methylmorpholine (2.87 mL, 26.07 mmol). N-(3-Dimethylaminopropyl)-N-ethylcarbodiimide hydrochloride (5.00 g, 26.07 mmol) was then added portionwise (5 portions) over 20 min. The mixture was stirred at 15 C. for 40 min. A 1M HCl solution (50 mL) was added. The mixture was extracted with DCM (225 mL). The organic layer was washed with a saturated solution of NaHCO.sub.3 (50 mL), H.sub.2O (50 mL), dried over Na.sub.2SO.sub.4 and concentrated in vacuo to provide the tert-butyl N-[(1R)-1-(hydroxymethyl)-2-[methoxy(methyl)amino]-2-oxo-ethyl]carbamate (10) (5.42 g, 21.83 mmol, 89%) as a white solid.

    [0273] .sup.1H NMR (400 MHz, CDCl.sub.3) 1.44 (s, 9H), 2.65 (s, 1H), 3.23 (s, 3H), 3.62-3.97 (m, 5H), 4.79 (s, 1H), 5.60 (d, J=8.4 Hz, 1H).

    Step 2: Preparation of Intermediate tert-butyl N-[(1R)-1-[[tert-butyl(dimethyl)silyl]oxymethyl]-2-[methoxy(methyl)amino]-2-oxo-ethyl]carbamate (11)

    [0274] To a solution of compound (10) (5.42 g, 21.8 mmol), imidazole (4.46 g, 65.5 mmol) and DMAP (133 mg, 1.1 mmol) in anhydrous DMF (17 mL) at rt was portionwise added tert-butyldimethylsilyl chloride (3.95 g, 26.2 mmol). The mixture was stirred for 2 h then poured in H.sub.2O (50 mL). The aqueous layer was extracted with EtOAc (240 mL). The organic layer was washed with 1 M HCl (50 mL), brine (40 mL), dried over Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (cyclohexane/EtOAc 100/0 to 50/50) to provide tert-butyl N-[(1R)-1-[[tert-butyl(dimethyl)silyl]oxymethyl]-2-[methoxy(methyl)amino]-2-oxo-ethyl]carbamate (11) (7.09 g, 19.5 mmol, 89%) as a colorless oil.

    [0275] MS m/z ([M+Na].sup.+) 385, ([M+H].sup.+) 363.

    [0276] .sup.1H NMR (400 MHz, CDCl.sub.3) 0.03 (s, 6H), 0.87 (s, 9H), 1.44 (s, 9H), 3.21 (s, 3H), 3.64-3.94 (m, 5H), 4.75 (s, 1H), 5.35 (d, J=9.0 Hz, 1H).

    Step 3: Preparation of Intermediate (2R)-2-amino-3-[tert-butyl(dimethyl)silyl]oxy-N-methoxy-N-methyl-propanamide (12)

    [0277] A solution of compound (11) (4.60 g, 12.69 mmol) and ZnBr.sub.2 (5.71 g, 25.38 mmol) in DCM (37 mL) was stirred at room temperature for 2 h30. A 2M NaOH solution (25 mL) was added followed by H.sub.2O (25 mL). The suspension was filtrated. The solid was washed with H.sub.2O and DCM. The filtrate was extracted with DCM. The organic layer was washed with brine, dried over Na.sub.2SO.sub.4 and concentrated in vacuo to provide (2R)-2-amino-3-[tert-butyl(dimethyl)silyl]oxy-N-methoxy-N-methyl-propanamide (12) (3.17 g, 12.08 mmol, 96%) as a colorless oil.

    [0278] MS m/z ([2M+H].sup.+) 525, ([M+H].sup.+) 263.

    [0279] .sup.1H NMR (400 MHz, CDCl.sub.3) 0.07 (s, 6H), 0.90 (s, 9H), 1.71 (bs, 2H), 3.23 (s, 3H), 3.63 (dd, J=9.6, 6.6 Hz, 1H), 3.74 (s, 3H), 3.81 (dd, J=9.6, 5.4 Hz, 1H), 3.87-3.96 (m, 1H).

    Step 4: Preparation of Intermediate tert-butyl N-allyl-N-[(1R)-1-[[tert-butyl(dimethyl)silyl]oxymethyl]-2-[methoxy(methyl)amino]-2-oxo-ethyl]carbamate (13)

    [0280] To a solution of compound (12) (3.17 g, 12.08 mmol) in anhydrous DMF (24 mL) at 0 C. was added K.sub.2CO.sub.3 (3.34 g, 24.16 mmol). The mixture was stirred at this temperature for 20 min before adding allyl bromide (1.15 mL, 13.29 mmol). The mixture was stirred for 1 h at 0 C. then 2 h at rt. Di-tert-butyl dicarbonate (3.95 g, 18.12 mmol) was added and the mixture maintained at rt overnight. H.sub.2O (50 mL) was added. The mixture was extracted with EtOAc (230 mL). The organic layer was washed with brine (40 mL), dried over Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (cyclohexane/EtOAc: 90/10) to provide tert-butyl N-allyl-N-[(1R)-1-[[tert-butyl(dimethyl)silyl]oxymethyl]-2-[methoxy(methyl)amino]-2-oxo-ethyl]carbamate (13) (2.06 g, 5.11 mmol, 42%) as a colorless oil.

    [0281] MS m/z ([M+Na].sup.+) 425, ([M+H].sup.+) 403.

    [0282] .sup.1H NMR (400 MHz, CDCl.sub.3) 0.05 (s, 6H), 0.87 (s, 9H), 1.38-1.60 (m, 9H), 3.16 (s, 3H), 3.73 (s, 3H), 3.78-4.07 (m, 4H), 4.87-5.40 (m, 3H), 5.68-5.96 (m, 1H).

    Step 5: Preparation of Intermediate tert-butyl N-allyl-N-[(1R)-1-[[tert-butyl(dimethyl)silyl]oxymethyl]-2-oxo-pent-3-enyl]carbamate (14)

    [0283] A solution of compound (13) (1.84 g, 4.57 mmol) in anhydrous THF (5 mL) was dropwise added to a propen-1-ylmagnesium bromide solution 0.5M in THF (18.3 mL, 9.14 mmol) at 0 C. under nitrogen atmosphere. The mixture was stired at 0 C. for 20 min. H.sub.2O (15 mL) and a saturated solution of NH.sub.4Cl (15 mL) were added. The mixture was extracted with tert-butyl methyl ether (220 mL). The organic layer was washed with 1M HCl (20 mL), dried over Na.sub.2SO.sub.4 and concentrated in vacuo to provide tert-butyl N-allyl-N-[(1R)-1-[[tert-butyl(dimethyl)silyl]oxymethyl]-2-oxo-pent-3-enyl]carbamate (14) (1.70 g, 4.43 mmol, 97%) which was used without further purification.

    [0284] MS m/z ([M+Na].sup.+) 406.

    Step 6: Preparation of Intermediate tert-butyl (2R)-2-[[tert-butyl(dimethyl)silyl]oxymethyl]-3-oxo-2,6-dihydropyridine-1-carboxylate (15)

    [0285] A solution of compound (14) (2.48 g, 6.47 mmol) and (1,3-Bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene)dichloro(o-isopropoxyphenylmethylene)ruthenium (203 mg, 0.32 mmol) in DCM was refluxed for 1 h. The mixture was concentrated in vacuo. The residue was purified by flash chromatography on silica gel (DCM/EtOAc: 100/0 to 95/5) to provide tert-butyl (2R)-2-[[tert-butyl(dimethyl)silyl]oxymethyl]-3-oxo-2,6-dihydropyridine-1-carboxylate (15) (2.07 g, 6.06 mmol, 93%) as a greenish solid.

    [0286] .sup.1H NMR (400 MHz, CDCl.sub.3) 0.04 (s, 3H), 0.01 (s, 3H), 0.82 (s, 9H), 1.47 (s, 6H), 1.50 (s, 3H), 3.70-4.19 (m, 3H), 4.38-4.75 (m, 2H), 6.18 (d, J=10.3, 1H), 6.82-7.08 (m, 1H).

    Step 7: Preparation of Intermediate tert-butyl (2R)-2-[[tert-butyl(dimethyl)silyl]oxymethyl]-3-hydroxy-3-thiazol-2-yl-2,6-dihydropyridine-1-carboxylate (16)

    [0287] To a solution of 2-bromothiazole (1.09 mL, 12.12 mmol) in anhydrous THF (12 mL) under nitrogen atmosphere at 0 C. was dropwise added a isopropylmagnesium chloride solution 2.0 M in THF (6.06 mL, 12.12 mmol). The mixture was stirred at 0 C. for 20 min then a solution of compound (15) (2.07 g, 6.06 mmol) in anhydrous THF (6 mL) was dropwise added. The mixture was stirred at rt for 45 min. H.sub.2O (15 mL) and a saturated solution of NH.sub.4Cl (15 mL) were added. The layers were separated. The aqueous layer was extracted with EtOAc (215 mL). The combined organic layers were washed with brine (20 mL), dried over Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (cyclohexane/EtOAc: 80/20 to 40/60) to provide tert-butyl (2R)-2-[[tert-butyl(dimethyl)silyl]oxymethyl]-3-hydroxy-3-thiazol-2-yl-2,6-dihydropyridine-1-carboxylate (16) (1.77 g, 4.15 mmol, 68%) as a brown oil.

    [0288] MS m/z ([M+H].sup.+) 427.

    Step 8: Preparation of Intermediate tert-butyl (6S)-3-bromo-6-[[tert-butyl(dimethyl)silyl]oxymethyl]-5-thiazol-2-yl-3,6-dihydro-2H-pyridine-1-carboxylate (17)

    [0289] Thionyl bromide (0.36 mL, 4.56 mmol) was dropwise added to a solution of TEA (0.64 mL, 4.56 mmol) and compound (16) (1.77 g, 4.15 mmol) in anhydrous DCM (18 mL) at 0 C. The mixture was stirred at 0 C. for 20 min then poured in a mixture of ice and H.sub.2O (50 mL). The layers were separated. The aqueous layer was extracted with DCM (220 mL). The combined organic layers were washed with brine (20 mL) dried over Na.sub.2SO.sub.4 and concentrated in vacuo to provide tert-butyl (6S)-3-bromo-6-[[tert-butyl(dimethyl)silyl]oxymethyl]-5-thiazol-2-yl-3,6-dihydro-2H-pyridine-1-carboxylate (17) (1.98 g, 4.04 mmol, 97%) as a brown oil which was used without further purification.

    [0290] .sup.1H NMR (300 MHz, CDCl.sub.3) 0.26-0.05 (m, 6H), 0.63-0.89 (m, 9H), 1.47-1.53 (m, 9H), 3.78-4.19 (m, 3H), 4.38-4.90 (m, 2H), 5.15-5.56 (m, 1H), 6.73-6.78 (m, 1H), 7.24-7.28 (m, 1H), 7.77-7.82 (m, 1H).

    Step 9: Preparation of Intermediate tert-butyl (3R, 6S)-3-(allyloxyamino)-6-[[tert-butyl(dimethyl)silyl]oxymethyl]-5-thiazol-2-yl-3,6-dihydro-2H-pyridine-1-carboxylate (18a) and tert-butyl (3S, 6S)-3-(allyloxyamino)-6-[[tert-butyl(dimethyl)silyl]oxymethyl]-5-thiazol-2-yl-3,6-dihydro-2H-pyridine-1-carboxylate (18b)

    [0291] To a suspension of NaH 60% in oil (202 mg, 5.06 mmol) in anhydrous DMF (6 mL) at 0 C. under nitrogen atmosphere was portionwise added N-allyloxy-2-nitro-benzenesulfonamide (1.31 g, 5.07 mmol). The mixture was stirred at 0 C. for 15 min then a solution of compound (17) (1.98 g, 4.04 mmol) in anhydrous DMF (6 mL) was dropwise added. The mixture was stirred for 90 min at 0 C. then H.sub.2O (20 mL) was added. The mixture was extracted with EtOAc (220 mL). The organic layer was washed with brine (20 mL), dried over Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (dichloromethane). The fractions containing the nosylated intermediate were combined and concentrated in vacuo. The residue was dissolved in ACN (30 mL) and K.sub.2CO.sub.3 (2.92 g, 21.14 mmol) and thiophenol (2.17 mL, 21.14 mmol) were added. The mixture was stirred at rt for 1 h then concentrated in vacuo. The residue was dissolved in EtOAc (20 mL), washed with a NaOH 2.0 M solution (20 mL), dried over Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (dichloromethane/EtOAc: 100/0 to 80/20) to provide compound (18b) (713 mg, 1.48 mmol) and a mixture (18b)/(18a) (30/70) (868 mg, 1.80 mmol) (yield: 81%).

    [0292] MS m/z ([M+H].sup.+) 482.

    [0293] (18a) (3R, 6S):

    [0294] .sup.1H NMR (400 MHz, CDCl.sub.3) 0.30-0.03 (m, 6H), 0.79 (s, 9H), 1.50 (s, 9H), 3.44 and 3.52 (dd, J=13.8, 3.4 Hz, 1H), 3.60-3.72 (m, 1H), 3.85-4.07 (m, 2H), 4.18-4.34 (m, 2H), 4.47 and 4.57 (d, J=13.8 Hz, 1H), 5.10-5.43 (m, 3H), 5.89-6.01 (m, 1H), 6.53 and 6.56 (d, J=5.3 Hz, 1H), 7.21 and 7.22 (d, J=3.3 Hz, 1H), 7.75 and 7.77 (d, J=3.3 Hz, 1H).

    [0295] (18b) (3S, 6S):

    [0296] .sup.1H NMR (400 MHz, CDCl.sub.3) 0.18-0.08 (m, 6H), 0.81 (s, 9H), 1.49 (s, 9H), 3.17 and 3.29 (t, J=11.4 Hz, 1H), 3.77-4.07 (m, 3H), 4.17-4.26 (m, 2H), 4.30 and 4.51 (dd, J=12.7, 6.3 Hz, 1H), 5.15-5.51 (m, 4H), 5.89-6.02 (m, 1H), 6.60 (s, 1H), 7.21 (s, 1H), 7.76 (s, 1H).

    Step 10: Preparation of Intermediate (2S,5R)-6-allyloxy-2-(hydroxymethyl)-3-thiazol-2-yl-1,6-diazabicyclo[3.2.1]oct-3-en-7-one (19a) and (2S,5S)-6-allyloxy-2-(hydroxymethyl)-3-thiazol-2-yl-1,6-diazabicyclo[3.2.1]oct-3-en-7-one (19b)

    [0297] To a solution of a mixture cis/trans (30/70) of compounds (18b/18a) (868 mg, 1.80 mmol) in anhydrous DCM (9 mL) at 0 C. under nitrogen were added TEA (0.50 mL, 3.60 mmol) and diphosgene (0.283 mL, 2.34 mmol). The mixture was stirred at 0 C. for 30 min, diluted with DCM (10 mL) and washed with brine (10 mL). The organic layer was dried over Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was dissolved in anhydrous dioxane (2 mL) and dropwise added to 4 M HCl solution in dioxane (9 mL). The mixture was stirred at rt for 1 h and concentrated in vacuo. The residue was dissolved in anhydrous dichloromethane (18 mL) cooled at 0 C. and triethylamine (1.0 mL, 7.21 mmol) was added. The mixture was stirred at rt for 15 min then washed with brine (10 mL). 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: 80/20 to 40/60) to provide (2S,5R)-6-allyloxy-2-(hydroxymethyl)-3-thiazol-2-yl-1,6-diazabicyclo[3.2.1]oct-3-en-7-one (19a) (290 mg, 0.99 mmol) and (2S,5S)-6-allyloxy-2-(hydroxymethyl)-3-thiazol-2-yl-1,6-diazabicyclo[3.2.1]oct-3-en-7-one (19b) (115 mg, 0.39 mmol) (yield: 76%).

    [0298] MS m/z ([M+H].sup.+) 294.

    [0299] (19a) (3R, 6S):

    [0300] .sup.1H NMR (400 MHz, CDCl.sub.3) 3.23-3.43 (m, 2H), 3.67 (bs, 1H), 3.94 (dd, J=11.4, 7.2 Hz, 1H), 3.99-4.06 (m, 1H), 4.20 (dd, J=11.6, 4.5 Hz, 1H), 4.32-4.48 (m, 2H), 4.52-4.64 (m, 1H), 5.29 (d, J=10.3 Hz, 1H), 5.34 (dd, J=17.2, 1.5 Hz, 1H), 5.85-6.10 (m, 1H), 6.98 (d, J=5.2 Hz, 1H), 7.23 (d, J=3.3 Hz, 1H), 7.71 (d, J=3.3, 1H).

    [0301] (19b) (3S, 6S):

    [0302] .sup.1H NMR (400 MHz, CDCl.sub.3) 3.09 (dd, J=14.1, 3.3 Hz, 1H), 3.72-3.85 (m, 1H), 4.00-4.17 (m, 1H), 4.20-4.40 (m, 3H), 4.86-5.03 (m, 2H), 5.19 (d, J=10.4 Hz, 1H), 5.30 (dd, J=17.3, 1.7 Hz, 1H), 5.45 (d, J=5.4 Hz, 1H), 5.84-6.03 (m, 1H), 6.55 (d, J=5.8 Hz, 1H), 7.30 (d, J=3.2 Hz, 1H), 7.74 (d, J=3.2 Hz, 1H).

    Step 11: Preparation of Intermediate [(2S,5R)-6-allyloxy-7-oxo-3-thiazol-2-yl-1,6-diazabicyclo[3.2.1]oct-3-en-2-yl]methyl methanesulfonate (20)

    [0303] To a solution of compound (19a) (290 mg, 0.989 mmol) in anhydrous DCM (3 mL) at 0 C. under atmosphere of nitrogen were successively added TEA (0.200 mL, 1.43 mmol) and MsCI (92 L, 1.18 mmol). The mixture was stirred at 0 C. H.sub.2O (5 mL) was added. The layers were separated. The aqueous layer was extracted with DCM (25 mL). The combined organic layers were washed with a saturated solution of NaHCO.sub.3 (10 mL), dried over Na.sub.2SO.sub.4 and concentrated in vacuo to provide [(2S,5R)-6-allyloxy-7-oxo-3-thiazol-2-yl-1,6-diazabicyclo[3.2.1]oct-3-en-2-yl]methyl methanesulfonate (20) (337 mg, 0.907 mmol, 92%) as an off white solid.

    [0304] MS m/z ([M+H].sup.+) 372.

    [0305] .sup.1H NMR (300 MHz, CDCl.sub.3) 3.00 (s, 3H), 3.40 (ddd, J=11.4, 2.6, 1.3 Hz, 1H), 3.50 (dd, J=11.4, 0.8 Hz, 1H), 4.00-4.10 (m, 1H), 4.34-4.52 (m, 2H), 4.85 (s, 3H), 5.27-5.41 (m, 2H), 5.93-6.10 (m, 1H), 7.07 (d, J=5.2 Hz, 1H), 7.26 (d, J=3.3 Hz, 1H), 7.75 (d, J=3.3 Hz, 1H).

    Step 12: Preparation of Intermediate tert-butyl N-[[(2S,5R)-6-allyloxy-7-oxo-3-thiazol-2-yl-1,6-diazabicyclo[3.2.1]oct-3-en-2-yl]methyl]carbamate (21)

    [0306] A mixture of compound (20) (337 mg, 0.907 mmol) and NaN.sub.3 (295 mg, 4.54 mmol) in anhydrous DMF (3.4 mL) was stirred at 65 C. for 20 h. The mixture was poured in H.sub.2O (10 mL) and extracted with ethyl acetate (210 mL). The organic layer was washed with brine (10 mL), dried over Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was dissolved in anhydrous THF (3 mL) and anhydrous toluene (3 mL) and cooled at 0 C. under nitrogen atmosphere. A trimethylphosphine solution 1M in THF (1.36 mL, 1.36 mmol) was dropwise added and the mixture was stirred at rt for 1 h. The mixture was cooled at 0 C. and a solution of 2-(Boc-oxyimino)-2-phenylacetonitrile (335 mg, 1.36 mmol) in anhydrous THF (2 mL) was added. The mixture was stirred at rt for 3 h. H.sub.2O (10 mL) was added and the layers separated. The aqueous layer was extracted with EtOAc (210 mL). The combined organic layers were washed with brine (10 mL), dried over Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (cyclohexane/EtOAc: 70/30 to 0/100) then by preparative TLC (cyclohexane/EtOAc: 50/50) to provide tert-butyl N-[[(2S,5R)-6-allyloxy-7-oxo-3-thiazol-2-yl-1,6-diazabicyclo[3.2.1]oct-3-en-2-yl]methyl]carbamate (21) (115 mg, 0.292 mmol, 32%) as a white solid.

    [0307] MS m/z ([M+H].sup.+) 393.

    [0308] .sup.1H NMR (300 MHz, CDCl.sub.3) 1.44 (s, 9H), 3.18-3.42 (m, 3H), 3.97-4.10 (m, 2H), 4.33-4.51 (m, 2H), 4.61 (ddd, J=11.0, 4.4, 1.4 Hz, 1H), 5.13 (s, 1H), 5.25-5.42 (m, 2H), 5.92-6.11 (m, 1H), 6.94-7.00 (m, 1H), 7.23 (d, J=3.2 Hz, 1H), 7.75 (d, J=3.2 Hz, 1H).

    Step 13: Preparation of Intermediate sodium [(2S,5R)-2-[(tert-butoxycarbonylamino)methyl]-7-oxo-3-thiazol-2-yl-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl]sulfate (22)

    [0309] To a solution of compound (21) (115 mg, 0.293 mmol) in anhydrous DCM (1 mL) under nitrogen atmosphere were successively added AcOH (34 L, 0.586 mmol) and Pd(PPh.sub.3).sub.4 (169 mg, 0.146 mmol). The mixture was stirred at rt for 1 h then concentrated in vacuo. The residue was purified by flash chromatography on silica gel (DCM/acetone: 100/0 to 0/100) to provide a mixture of expected intermediate and triphenylphosphine oxide. The mixture was dissolved in pyridine (2 mL) and sulfur trioxide trimethylamine complex (417 mg, 3.00 mmol) was added. The mixture was stirred at rt overnight then concentrated in vacuo. DCM (5 mL) was added to the residue and the precipitate filtered. The filtrate was concentrated and the residue purified by flash chromatography on silica gel (DCM/acetone: 60/40 to 0/100). The fractions containing the expected intermediate were combined and concentrated in vacuo. The residue was dissolved in H.sub.2O (1 mL) and converted after ion exchange (Dowex sodium form column) to sodium [(2S,5R)-2-[(tert-butoxycarbonylamino)methyl]-7-oxo-3-thiazol-2-yl-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl]sulfate (22) (38 mg, 0.083 mmol, 29%) as a white solid.

    [0310] MS m/z ([M+H].sup.+) 433.

    [0311] MS m/z ([MH].sup.) 431.

    [0312] .sup.1H NMR (400 MHz, D.sub.2O) 1.39 (s, 9H), 3.34-3.68 (m, 5H), 4.43-4.54 (m, 2H), 7.03 (d, J=5.1 Hz, 1H), 7.52 (d, J=3.4 Hz, 1H), 7.75 (d, J=3.4 Hz, 1H).

    Step 14: Preparation of sodium and 2,2,2-trifluoroacetate [(2S,5R)-2-(azaniumylmethyl)-7-oxo-3-thiazol-2-yl-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl] sulfate (Example 2)

    [0313] A solution of compound (22) (38 mg, 0.083 mmol) in anhydrous DCM (0.67 mL) was added to a mixture of DCM (1 mL) and TFA (1 mL) at 0 C. The mixture was stirred at 0 C. for 30 min then concentrated in vacuo. The residue was co-evaporated three times with DCM (3 mL). The residue was dissolved in H.sub.2O (2 mL) and lyophilized to provide sodium and 2,2,2-trifluoroacetate disalt of [(2S,5R)-2-(azaniumylmethyl)-7-oxo-3-thiazol-2-yl-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl] sulfate (Example 2) (35 mg, 0.074 mmol, 89%) as an off white solid.

    [0314] MS m/z ([M+H].sup.+) 333.

    [0315] MS m/z ([MH].sup.) 331.

    [0316] .sup.1H NMR (300 MHz, D.sub.2O) 3.36 (dd, J=13.8, 11.4 Hz, 1H), 3.54 (d, J=1.6 Hz, 2H), 3.65 (dd, J=13.8, 3.9 Hz, 1H), 4.53 (dt, J=5.2, 1.6 Hz, 1H), 4.72 (ddd, J=11.4, 3.9, 1.6 Hz, 1H), 7.16 (dd, J=5.2, 1.6 Hz, 1H), 7.53 (d, J=3.3 Hz, 1H), 7.77 (d, J=3.3 Hz, 1H).

    Example 3: synthesis of lithium and 2,2,2-trifluoroacetate [trans-2-(azaniumylmethyl)-4-oxazol-5-yl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl]oxyl-2,2-difluoro-acetate

    [0317] ##STR00032##

    Step 1: Preparation of Intermediate tert-butyl N-[[trans-6-hydroxy-4-oxazol-5-yl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-2-yl]methyl]carbamate (23)

    [0318] To a solution of tert-butyl N-[[trans-6-allyloxy-4-oxazol-5-yl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-2-yl]methyl]carbamate (7) (200 mg, 0.53 mmol) and glacial acetic acid (49 L, 0.85 mmol) in anhydrous DCM (5.3 mL) was added in one portion Pd(PPh.sub.3).sub.4 (307 mg, 0.27 mmol). The mixture was stirred at rt for 30 min and concentrated under argon flow. The crude was purified by flash chromatography on silica gel (petroleum ether/acetone 100/0 to 40/60) to give tert-butyl N-[trans-6-hydroxy-4-oxazol-5-yl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-2-yl]methyl]carbamate (23) (176 mg, 0.52 mmol, 90.5%).

    [0319] MS m/z ([M+H].sup.+) 337.

    [0320] .sup.1H NMR (400 MHz, CDCl.sub.3): (ppm) 1.46 (s, 9H), 3.14-3.20 (m, 1H), 3.28 (d, J=11.2 Hz, 1H), 3.42 (dd, J=11.2/2.6 Hz, 1H), 3.58-3.64 (m, 1H), 4.01-4.05 (m, 1H), 4.15-4.16 (m, 1H), 5.10-5.11 (m, 1H), 5.97-5.98 (m, 1H), 7.11 (s, 1H), 7.79 (s, 1H).

    Step 2: Preparation of Intermediate ethyl 2-[trans-2-[(tert-butoxycarbonylamino)methyl]-4-oxazol-5-yl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl]oxy]-2,2-difluoro-acetate (24)

    [0321] tert-butyl N-[trans-6-hydroxy-4-oxazol-5-yl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-2-yl]methyl]carbamate (23) (161.5 mg, 0.48 mmol) was solubilized in DMF (5.30 mL) at 20 C. with DBU (80 L, 0.53 mmol) and ethyl 2-bromo-2,2-difluoro-acetate (308 L, 2.40 mmol). The reaction was stirred for 1h15 at 20 C. Water was added and the mixture was extracted twice with EtOAc. The organic layer was dried over Na.sub.2SO.sub.4, filtered, and concentrated in vacuo. The crude was purified by flash chromatography on silica gel (petroleum ether/acetone 100/0 to 60/40) to give ethyl 2-[trans-2-[(tert-butoxycarbonylamino)methyl]-4-oxazol-5-yl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl]oxy]-2,2-difluoro-acetate

    [0322] (24) (179 mg, 0.39 mmol, 81%).

    [0323] MS m/z ([M+H].sup.+) 459.

    [0324] .sup.1H NMR (400 MHz, CDCl.sub.3): (ppm) 1.32 (t, J=7.1 Hz, 3H), 1.46 (s, 9H), 3.21-3.29 (m, 1H), 3.37-3.39 (m, 1H), 3.46-3.50 (m, 1H), 3.58-3.64 (m, 1H), 4.11-4.15 (m, 1H), 4.34 (q, J=7.1 Hz, 2H), 4.37-4.38 (m, 1H), 5.0-5.01 (bs, 1H), 6.05-6.06 (m, 1H), 7.15 (s, 1H), 7.84 (s, 1H).

    Step 3: Preparation of Intermediate lithium salt of trans-2-[[2-[(tert-butoxycarbonylamino)methyl]-4-oxazol-5-yl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl]oxy]-2,2-difluoro-acetate (25)

    [0325] Ethyl 2-[trans-2-[(tert-butoxycarbonylamino)methyl]-4-oxazol-5-yl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl]oxy]-2,2-difluoro-acetate (24) (79 mg, 0.17 mmol) was solubilized in THF (1 mL) and water (0.31 mL) at 0 C. A solution of 1N LiOH (215 L, 0.21 mmol) was then dropped. The mixture was stirred for 30 min at 0 C. The reaction mixture was acidified with 0.1N HCl (50 L) and concentrated to remove THF. The resulting aqueous layer was frozen and lyophilized. The resulting salt was triturated with Et.sub.2O to provide lithium trans-2-[[2-[(tert-butoxycarbonylamino)methyl]-4-oxazol-5-yl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl]oxy]-2,2-difluoro-acetate (25) (60 mg, 0.14 mmol, 80%) as a white solid.

    [0326] MS m/z ([M+H].sup.+) 431.

    [0327] .sup.1H NMR (400 MHz, CDCl.sub.3): (ppm) 1.39 (s, 9H), 3.19-3.28 (m, 3H), 3.37-3.40 (m, 1H), 3.86-3.91 (m, 1H), 4.52-4.53 (m, 1H), 6.0-6.01 (m, 1H), 7.45 (s, 1H), 8.37 (s, 1H).

    Step 4: Preparation of lithium and 2,2,2-trifluoroacetate [trans-2-(azaniumylmethyl)-4-oxazol-5-yl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl]oxy]-2,2-difluoro-acetate (Example 3)

    [0328] At 0 C. TFA (6.6 L) was slowly added to a solution of compound (25) (25 mg, 0.057 mmol) in anhydrous DCM (1 mL). After 2 h at 0 C. an excess of TFA (600 L) was added. The mixture was stirred at 0 C. for 1 h more and concentrated in vacuo. The residue was dissolved in H.sub.2O (100 L) and lyophilized to provide lithium and 2,2,2-trifluoroacetate [trans-2-(azaniumylmethyl)-4-oxazol-5-yl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl]oxy]-2,2-difluoro-acetate (Example 3) (26 mg, 0.057 mmol, 100%).

    [0329] MS m/z ([M+H].sup.+) 331.

    [0330] MS m/z ([MH].sup.) 329.

    [0331] .sup.1H NMR (400 MHz, DMSO-d.sub.6) 3.22-3.63 (m, 4H), 4.17-4.22 (m, 1H), 4.66-4.67 (m, 1H), 6.11-6.12 (m, 1H), 7.33 (s, 1H), 8.06 (bs, 3H), 8.44 (s, 1H).

    Example 4: synthesis of sodium [trans-2-(methoxymethyl)-4-oxazol-5-yl-1,6-diazabicyclo[3.2.1]oct-3-en-7-one] sulfate

    [0332] ##STR00033##

    Step 1: Preparation of Intermediate trans-6-allyloxy-2-(methoxymethyl)-4-oxazol-5-yl-1,6-diazabicyclo[3.2.1]oct-3-en-7-one (26)

    [0333] A solution of trans-6-allyloxy-2-(hydroxymethyl)-4-oxazol-5-yl-1,6-diazabicyclo[3.2.1]oct-3-en-7-one (6) (140 mg, 0.50 mmol) in anhydrous DMF (1.5 mL) was cooled to 0 C. Iodomethane (94 L, 1.51 mmol) followed by NaH 60% in oil (24 mg, 0.61 mmol) were added and the reaction mixture was stirred at 0 C. for 15 min. Water (2 mL) was added carefully and the mixture was extracted with EtOAc (23 mL). The organic layer was dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The crude was purified by flash chromatography on silica gel (cyclohexane/EtOAc 100/0 to 20/80) to provide trans-6-allyloxy-2-(methoxymethyl)-4-oxazol-5-yl-1,6-diazabicyclo[3.2.1]oct-3-en-7-one (26) (60 mg, 0.21 mmol, 41%).

    [0334] MS m/z ([M+H].sup.+) 292.

    [0335] .sup.1H NMR (300 MHz, CDCl.sub.3): (ppm) 3.36-3.43 (m, 4H), 3.50 (dd, J=11.2, 0.8 Hz, 1H), 3.63-3.75 (m, 2H), 4.11-4.19 (m, 2H), 4.33-4.49 (m, 2H), 5.27-5.39 (m, 2H), 5.91-6.06 (m, 1H), 6.10 (d, J=3.2 Hz, 1H), 7.03 (s, 1H), 7.82 (s, 1H).

    Step 2: Preparation of sodium [trans-2-(methoxymethyl)-4-oxazol-5-yl-1,6-diazabicyclo[3.2.1]oct-3-en-7-one] sulfate (example 4)

    [0336] To a solution of trans-6-allyloxy-2-(methoxymethyl)-4-oxazol-5-yl-1,6-diazabicyclo[3.2.1]oct-3-en-7-one (26) (60 mg, 0.206 mmol) and glacial acetic acid (24 L, 0.412 mmol) in anhydrous DCM (2.3 mL) was added in one portion Pd(PPh.sub.3).sub.4 (119 mg, 0.103 mmo). After stirring for 2 h, a solution of sulfur trioxide pyridine complex (133 mg, 0.834 mmol) in anhydrous pyridine (2.6 mL) was added and the resulting mixture was stirred overnight. The mixture was concentrated in vacuo, diluted with DCM and the precipitate filtered. The filtrate was concentrated in vacuo. The crude was purified by flash chromatography on silica gel (DCM/acetone 100/0 to 0/100). The fractions containing the expected intermediate were combined and concentrated in vacuo. The residue, dissolved in a mixture of H.sub.20/THF 7/3 (0.5 mL), 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 H.sub.2O). The fractions containing the desired compound were combined and concentrated in vacuo. The product was dissolved in a minimum amount of water, freezed and lyophilized to afford sodium [trans-2-(methoxymethyl)-4-oxazol-5-yl-1,6-diazabicyclo[3.2.1]oct-3-en-7-one] sulfate (example 4) (18 mg, 0.05 mmol, 24%).

    [0337] .sup.1H NMR (300 MHz, DMSO-d.sub.6): (ppm) 3.24-3.32 (m, 4H), 3.46 (d, J=11.4 Hz, 1H), 3.57-3.69 (m, 2H), 3.91-3.98 (m, 1H), 4.50-4.54 (m, 1H), 5.99 (d, J=2.9 Hz, 1H), 7.25 (s, 1H), 8.37 (s, 1H).

    Example 5: synthesis of sodium and 2,2,2-trifluoroacetate [(2S,5R)-2-(azaniumylmethyl)-7-oxo-3-oxazol-2-yl-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl] sulfate

    [0338] ##STR00034##

    Step 1: Preparation of Intermediate tert-butyl (2R)-2-[[tert-butyl(dimethyl)silyl]oxymethyl]-3-hydroxy-3-oxazol-2-yl-2,6-dihydropyridine-1-carboxylate (27)

    [0339] To a solution of borane tetrahydrofuran complex solution 1.0 M in THF (19 mL, 19 mmol) under nitrogen atmosphere at rt, was dropwise added oxazole (1.24 mL, 18.89 mmol). The mixture was stirred at rt for 1 h then cooled down to 78 C. A n-butyllithium solution 1.6 M in hexanes (12.2 ml, 19.5 mmol) was dropwise added and the mixture maintained at this temperature for 30 min. A solution of compound (15) (4.30 g, 12.6 mmol) in anhydrous THF (9 mL) was dropwise added. The mixture was stirred at 78 C. for 90 min. Ethanol containing 5% AcOH (30 mL) was added and the mixture was stirred at rt for 18 h. Water (50 mL) was added. The aqueous layer was extracted with EtOAc (250 mL). The combined organic layers were washed with a saturated solution of NaHCO.sub.3 (50 mL), dried over Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (cyclohexane/EtOAc: 80/20 to 40/60) to provide compound (27) (1.12 g, 2.72 mmol, 21%) as a cis/trans mixture.

    [0340] MS m/z ([M+H].sup.+) 411.

    [0341] 1H NMR (400 MHz, CDCl.sub.3) 0.10 and 0.11 (s, 6H), 0.90 (s, 9H), 1.33 and 1.37 (s, 9H), 3.40-3.78 (m, 2H), 4.00-4.08 (m, 1H), 4.26 and 4.38 (d, J=19.4 Hz, 1H), 4.68 and 4.99 (bs, 1H), 4.88 and 4.74 (t, J=7.2 Hz, 1H), 5.86-6.04 (m, 2H), 7.01 and 7.07 (s, 1H), 7.61 (d, J=0.8 Hz, 1H).

    Step 2: Preparation of Intermediate tert-butyl (6S)-3-bromo-6-[[tert-butyl(dimethyl)silyl]oxymethyl]-5-oxazol-2-yl-3,6-dihydro-2H-pyridine-1-carboxylate (28)

    [0342] Using the procedure described in example 2 (step 8) the intermediate (27) (1.12 g, 2.73 mmol) is converted into intermediate (28) (1.25 g, 2.64 mmol, 96%) as a cis/trans mixture which was used without further purification.

    Step 3: Preparation of Intermediate tert-butyl (6S)-3-(allyloxyamino)-6-[[tert-butyl(dimethyl)silyl]oxymethyl]-5-oxazol-2-yl-3,6-dihydro-2H-pyridine-1-carboxylate (29)

    [0343] Using the procedure described in example 2 (step 9) the intermediate (28) (1.25 g, 2.64 mmol) is converted into intermediate (29) (750 mg, 1.61 mmol, 61%) as a mixture cis/trans (62/38) after purification by flash chromatography on silica gel (DCM/EtOAc from 100/0 to 70/30).

    [0344] MS m/z ([M+H].sup.+) 466.

    Step 4: Preparation of intermediates (2S,5R)-6-allyloxy-2-(hydroxymethyl)-3-oxazol-2-yl-1,6-diazabicyclo[3.2.1]oct-3-en-7-one (30a) and (2S,5S)-6-allyloxy-2-(hydroxymethyl)-3-oxazol-2-yl-1,6-diazabicyclo[3.2.1]oct-3-en-7-one (30b)

    [0345] Using the procedure described in example 2 (step 10) the intermediate (29) (750 mg, 1.61 mmol) is converted into intermediate (30a) (88 mg, 0.31 mmol, 20%) and intermediate (30b) (201 mg, 0.72 mmol, 45%) after purification and separation by preparative TLCs on silica gel (EtOAc).

    [0346] MS m/z ([M+H].sup.+) 278.

    [0347] 30a:

    [0348] 1H NMR (400 MHz, CDCl.sub.3) 3.33 (s, 2H), 3.51 (bs, 1H), 3.93 (dd, J=11.4, 7.5 Hz, 1H), 4.03-4.08 (m, 1H), 4.22 (dd, J=11.6, 4.7 Hz, 1H), 4.33-4.52 (m, 3H), 5.24-5.40 (m, 2H), 5.92-6.07 (m, 1H), 7.11 (s, 1H), 7.19 (d, J=5.3 Hz, 1H), 7.58 (s, 1H).

    [0349] 30b:

    [0350] .sup.1H NMR (400 MHz, CDCl.sub.3) 3.19 (dd, J=14.1, 3.3 Hz, 1H), 3.90 (bs, 1H), 4.21 (t, J=8.5 Hz, 1H), 4.32-4.47 (m, 3H), 4.86-4.98 (m, 1H), 5.04 (t, J=9.0 Hz, 1H), 5.30 (dd, J=10.4, 1.0 Hz, 1H), 5.35-5.45 (m, 1H), 5.55 (s, 1H), 5.99-6.09 (m, 1H), 6.86 (d, J=5.9 Hz, 1H), 7.26 (d, J=0.8 Hz, 1H), 7.76 (d, J=0.8 Hz, 1H).

    Step 5: Preparation of Intermediate [(2S,5R)-6-allyloxy-7-oxo-3-oxazol-2-yl-1,6-diazabicyclo[3.2.1]oct-3-en-2-yl]methyl methanesulfonate (31)

    [0351] Using the procedure described in example 2 (step 11) the intermediate (30a) (119 mg, 0.43 mmol) is converted into intermediate (31) (153 mg, 0.43 mmol, 100%) as a yellow oil which was used without further purification.

    [0352] MS m/z ([M+H].sup.+) 356.

    [0353] 1H NMR (400 MHz, CDCl.sub.3) 3.05 (s, 3H), 3.40 (ddd, J=11.5, 2.7, 1.3 Hz, 1H), 3.48 (dd, J=11.5, 0.8 Hz, 1H), 4.07 (dd, J=5.2, 1.9 Hz, 1H), 4.35-4.50 (m, 2H), 4.66 (ddd, J=6.4, 3.9, 1.8 Hz, 1H), 4.85-4.89 (m, 2H), 5.29-5.41 (m, 2H), 5.95-6.07 (m, 1H), 7.14 (d, J=0.8 Hz, 1H), 7.27 (dt, J=5.2, 1.4 Hz, 1H), 7.59 (d, J=0.8 Hz, 1H).

    Step 6: Preparation of Intermediate tert-butyl N-[[(2S,5R)-6-allyloxy-7-oxo-3-oxazol-2-yl-1,6-diazabicyclo[3.2.1]oct-3-en-2-yl]methyl]carbamate (32)

    [0354] Using the procedure described in example 2 (step 12) the intermediate (31) (153 mg, 0.43 mmol) is converted into intermediate (32) (57 mg, 0.15 mmol, 35%) after purification by flash chromatography on silica gel (cyclohexane/EtOAc from 70/30 to 0/100) then by preparative TLCs on silica gel (cyclohexane/acetone 60/40).

    [0355] MS m/z ([M+H].sup.+) 377.

    [0356] .sup.1H NMR (400 MHz, CDCl.sub.3) 1.43 (s, 9H), 3.19-3.37 (m, 3H), 3.97-4.10 (m, 2H), 4.32-4.49 (m, 3H), 5.15 (bs, 1H), 5.24-5.40 (m, 2H), 5.89-6.07 (m, 1H), 7.11 (s, 1H), 7.16 (d, J=4.9, 1H), 7.56 (s, 1H).

    Step 7: Preparation of Intermediate sodium [(2S,5R)-2-[(tert-butoxycarbonylamino)methyl]-7-oxo-3-oxazol-2-yl-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl] sulfate (33)

    [0357] Using the procedure described in example 2 (step 13) the intermediate (32) (57 mg, 0.15 mmol) is converted into intermediate (33) (28 mg, 0.064 mmol, 42%).

    [0358] MS m/z ([M+H].sup.+) 417.

    [0359] MS m/z ([MH].sup.) 415.

    [0360] .sup.1H NMR (400 MHz, D.sub.2O) 1.43 (s, 9H), 3.41-3.58 (m, 2H), 3.64 (d, J=11.9 Hz, 1H), 3.74 (dd, J=14.8, 3.2 Hz, 1H), 4.40 (dd, J=9.9, 3.7 Hz, 1H), 4.53 (dd, J=5.3, 2.5 Hz, 1H), 7.22 (s, 1H), 7.27 (d, J=5.2 Hz, 1H), 7.85 (s, 1H).

    Step 8: Preparation of sodium and 2,2,2-trifluoroacetate [(2S,5R)-2-(azaniumylmethyl)-7-oxo-3-oxazol-2-yl-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl] sulfate (Example 5)

    [0361] Using the procedure described in example 2 (step 14) the intermediate (33) (28 mg, 0.064 mmol) is converted into Example 5 (22.8 mg, 0.050 mmol, 78%) as an off white solid.

    [0362] MS m/z ([M+H].sup.+) 317.

    [0363] MS m/z ([MH].sup.) 315.

    [0364] .sup.1H NMR (300 MHz, D.sub.2O) 3.41 (dd, J=13.5, 11.9 Hz, 1H), 3.57 (d, J=1.6 Hz, 2H), 3.79 (dd, J=13.8, 3.9 Hz, 1H), 4.54-4.66 (m, 2H), 7.24 (d, J=0.9 Hz, 1H), 7.37 (dd, J=5.3, 1.6 Hz, 1H), 7.86 (d, J=0.9 Hz, 1H).

    Example 6: synthesis of [trans-2-(aminomethyl)-4-(1-methylpyrimidin-2(1H)-one-5-yl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl] hydrogen sulfate

    [0365] ##STR00035##

    Step 1: Preparation of Intermediate trans-6-allyloxy-2-(hydroxymethyl)-4-iodo-1,6-diazabicyclo[3.2.1]oct-3-en-7-one (34)

    [0366] Using the procedure described in example 1 (step 5) the intermediate (4) (514 mg, 1.14 mmol) is converted into intermediate (34) (384 mg, 1.14 mmol, quantitative yield) as a brown oil which was used without further purification.

    [0367] MS m/z ([M+H].sup.+) 337.

    Step 2: Preparation of Intermediate tert-butyl N-[[trans-6-allyloxy-4-iodo-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-2-yl]methyl]carbamate (35)

    [0368] A solution of trans-6-allyloxy-2-(hydroxymethyl)-4-iodo-1,6-diazabicyclo[3.2.1]oct-3-en-7-one (34) (384 mg, 1.14 mmol) in DCM (11 mL) was cooled to 0 C. TEA (0.95 mL, 6.85 mmol) and MsCI (0.44 mL, 5.71 mmol) were added and the reaction mixture was stirred at the same temperature for 1 h. After completion, the reacting mixture was concentrated in vacuo. The crude was dissolved in DMF (11 mL) and NaN.sub.3 (371 mg, 5.71 mmol) was added. The reaction mixture was heated at 65 C. overnight and concentrated in vacuo. The crude was dissolved in a mixture of THF and toluene (3.8 mL/3.8 mL) and PMe.sub.3 (1M in tetrahydrofuran) (1.71 mL, 1.71 mmol) was added at 0 C. After 1 h stirring at rt, the mixture was cooled to 0 C. and a solution of BocON (422 mg, 1.71 mmol) in THF (3.8 mL) was dropwise added. The mixture was stirred overnight at rt and concentrated in vacuo. The crude was purified by flash chromatography on silica gel (DCM/Acetone from 95/5 to 0/100) to give tert-butyl N-[[trans-6-allyloxy-4-iodo-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-2-yl]methyl]carbamate (35) (43 mg, 0.10 mmol, 10% over 4 steps).

    [0369] MS m/z ([M+H].sup.+) 436.

    [0370] .sup.1H NMR (400 MHz, CDCl.sub.3): (ppm) 1.42 (s, 9H), 3.17-3.20 (m, 1H), 3.26-3.31 (m, 1H), 3.45-3.51 (m, 1H), 3.85-3.88 (m, 1H), 4.09 (d, J=2.0 Hz, 1H), 4.37-4.50 (m, 3H), 5.76-5.86 (m, 1H), 5.98-6.08 (m, 2H), 6.26 (d, J=2.0 Hz, 1H), 6.98 (bs, 1H).

    Step 3: Preparation of Intermediate tert-butyl N-[[trans-6-allyloxy-4-(1-methylpyrimidin-2(1H)-one-5-yl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-2-yl]methyl]carbamate (36)

    [0371] In a sealed flask, a mixture of tert-butyl N-[[trans-6-allyloxy-4-iodo-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-2-yl]methyl]carbamate (35) (43 mg, 0.10 mmol) and 5-trimethyltin-1-methylpyrimidin-2(1H)-one (32 mg, 0.12 mmol) in anhydrous THF (1.0 mL) was degassed under argon for 5 min before Pd(PPh.sub.3).sub.4 (6 mg, 0.005 mmol) and CuI (2 mg, 0.01 mmol) were added. The mixture was heated at 60 C. overnight. After completion, the reacting mixture was concentrated in vacuo. The crude was purified by flash chromatography on silica gel (DCM/Acetone from 70/30 to 0/100) to give tert-butyl N-[[trans-6-allyloxy-4-(1-methylpyrimidin-2(1H)-one-5-yl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-2-yl]methyl]carbamate (36) (12 mg, 0.03 mmol, 29%).

    [0372] MS m/z ([M+H].sup.+) 418.

    [0373] .sup.1H NMR (400 MHz, (CD.sub.3).sub.2CO): (ppm) 1.41 (s, 9H), 3.25-3.41 (m, 2H), 3.52 (s, 3H), 3.84-3.93 (m, 2H), 4.49-4.52 (m, 1H), 5.25 (d, J=10.4 Hz, 1H), 5.38 (dd, J=17.2, 1.2 Hz, 1H), 5.96 (d, J=2.0 Hz, 1H), 5.99-6.09 (m, 2H), 6.20-6.24 (m, 1H), 6.38 (dd, J=6.4, 4.0 Hz, 1H), 8.14 (dd, J=6.4, 2.8 Hz, 1H), 8.53 (d, J=2.8 Hz, 1H).

    Step 4: Preparation of Intermediate sodium [trans-2-[(tert-butoxycarbonylamino)methyl]-4-(1-methylpyrimidin-2(1H)-one-5-yl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl] sulfate (37)

    [0374] To a solution of tert-butyl N-[[trans-6-allyloxy-4-(1-methylpyrimidin-2(1H)-one-5-yl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-2-yl]methyl]carbamate (36) (12 mg, 0.03 mmol) and glacial AcOH (3 L, 0.06 mmol) in anhydrous DCM (0.14 mL) was added in one portion Pd(PPh.sub.3).sub.4 (17 mg, 0.014 mmol). After stirring for 2 h, a solution of sulfur trioxide pyridine complex (23 mg, 0.14 mmol) in dry pyridine (0.14 mL) was added and the resulting mixture was stirred overnight at 40 C. in the dark. The reaction mixture was concentrated in vacuo, diluted with DCM and filtered. The filtrate was concentrated in vacuo. The crude was dissolved in a mixture of H.sub.20/MeCN 7/3 (1 mL) 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 H.sub.2O). The fractions containing the desired compound were combined and concentrated in vacuo to give sodium [trans-2-[(tert-butoxycarbonylamino)methyl]-4-(1-methylpyrimidin-2(1H)-one-5-yl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl] sulfate (37) (15 mg).

    [0375] MS m/z ([M+H].sup.+) 458.

    [0376] MS m/z ([MH].sup.) 456.

    Step 5: Preparation of [trans-2-(aminomethyl)-4-(1-methylpyrimidin-2(1H)-one-5-yl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl] hydrogen sulfate (Example 6)

    [0377] The crude sodium [trans-2-[(tert-butoxycarbonylamino)methyl]-4-(1-methylpyrimidin-2(1H)-one-5-yl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl] sulfate (37) (14 mg, 0.03 mmol) was dissolved in DCM (1.0 mL), cooled to 0 C., and a pre-cooled solution of TFA (0.67 mL, 8.76 mmol) in DCM (0.67 mL) was dropwise added. After 1 h stirring at the same temperature, the reaction mixture was poured into DCM and concentrated in vacuo (co-evaporations with DCM). The solid residue was washed with DCM twice, solubilized into water, filtered through Isodisc and lyophilized to afford the crude sodium and 2,2,2-trifluoroacetate [trans-2-(azaniumylmethyl)-4-(1-methylpyrimidin-2(1H)-one-5-yl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl] sulfate. The crude sodium and 2,2,2-trifluoroacetate [trans-2-(azaniumylmethyl)-4-(1-methylpyrimidin-2(1H)-one-5-yl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl] sulfate (7.1 mg, 0.014 mmol) was then triturated 3 times in propan-2-ol and dried under reduced pressure to afford [trans-2-(aminomethyl)-4-(1-methylpyrimidin-2(1H)-one-5-yl)-7-oxo-1,6-diazabicyclo [3.2.1]oct-3-en-6-yl] sulfate (Example 6) (5.2 mg, 0.01 mmol, 35% over 4 steps).

    [0378] MS m/z ([M+H].sup.+) 358.

    [0379] MS m/z ([MH].sup.) 356.

    [0380] .sup.1H NMR (400 MHz, D.sub.2O): (ppm) 3.27-3.43 (m, 2H), 3.52 (d, J=11.6 Hz, 1H), 3.59 (dd, J=11.6, 2.8 Hz, 1H), 3.60 (s, 3H), 4.22-4.27 (ddd, J=11.6, 3.2, 2.8 Hz, 1H), 4.67 (d, J=2.8 Hz, 1H), 5.99 (d, J=2.8 Hz, 1H), 8.23 (d, J=3.2 Hz, 1H), 8.72 (d, J=3.2 Hz, 1H).

    Example 7: synthesis of [(2S,5R)-2-(quanidinomethyl)-7-oxo-3-thiazol-2-yl-1,6-diazabicyclo [3.2.1]oct-3-en-6-yl] hydrogen sulfate

    [0381] ##STR00036##

    Step 1: Preparation of Intermediate tert-butyl N[N-[[(2S,5R)-6-allyloxy-7-oxo-3-thiazol-2-yl-1,6-diazabicyclo[3.2.1]oct-3-en-2-yl]methyl]-N-tert-butoxycarbonyl-carbamimidoyl]carbamate (38)

    [0382] A mixture of compound (20) (150 mg, 0.404 mmol) and NaN.sub.3 (131 mg, 2.02 mmol) in anhydrous DMF (2.0 mL) was stirred at 65 C. for 24 h. The mixture was poured in H.sub.2O (5 mL) and extracted with EtOAc (25 mL). The organic layer was washed with brine (5 mL), dried over Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was dissolved in anhydrous THF (1 mL) and anhydrous toluene (1 mL) and cooled at 0 C. under nitrogen atmosphere. A PMe.sub.3 solution 1M in THF (0.52 mL, 0.518 mmol) was dropwise added and the mixture was stirred at rt for 1 h. The mixture was cooled at 0 C. and a solution of BocON (161 mg, 0.518 mmol) in anhydrous THF (0.7 mL) was added. The mixture was stirred at rt for 1 h. Water (5 mL) was added and the layers separated. The aqueous layer was extracted with EtOAc (25 mL). The combined organic layers were washed with brine (5 mL), dried over Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (cyclohexane/EtOAc: 95/5 to 0/100) to provide tert-butyl N[N-[[(2S,5R)-6-allyloxy-7-oxo-3-thiazol-2-yl-1,6-diazabicyclo[3.2.1]oct-3-en-2-yl]methyl]-N-tert-butoxycarbonyl-carbamimidoyl]carbamate (38) (40 mg, 0.075 mmol, 22%) as a white solid.

    [0383] MS m/z ([M+H].sup.+) 535.

    [0384] .sup.1H NMR (400 MHz, CDCl.sub.3) 1.47 (s, 9H), 1.50 (s, 9H), 3.29-3.36 (m, 1H), 3.40 (d, J=11.4 Hz, 1H), 3.65 (ddd, J=14.1, 11.4, 4.3 Hz, 1H), 4.02 (dd, J=5.0, 2.3 Hz, 1H), 4.19 (dt, J=14.2, 4.8 Hz, 1H), 4.35-4.48 (m, 2H), 4.76 (ddd, J=11.4, 4.3, 1.3 Hz, 1H), 5.27-5.38 (m, 2H), 5.95-6.08 (m, 1H), 6.96 (d, J=5.1 Hz, 1H), 7.21 (d, J=3.2 Hz, 1H), 7.74 (d, J=3.2 Hz, 1H), 8.80 (s, 1H), 11.43 (s, 1H).

    Step 2: Preparation of Intermediate sodium [(2S,5R)-2-[[bis(tert-butoxycarbonylamino)methyleneamino]methyl]-7-oxo-3-thiazol-2-yl-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl] sulfate (39)

    [0385] To a solution of tert-butyl N[N-[[(2S,5R)-6-allyloxy-7-oxo-3-thiazol-2-yl-1,6-diazabicyclo[3.2.1]oct-3-en-2-yl]methyl]-N-tert-butoxycarbonyl-carbamimidoyl]carbamate (38) (40 mg, 0.075 mmol) and glacial AcOH (9 L, 0.150 mmol) in anhydrous DCM (0.3 mL) was added in one portion Pd(PPh.sub.3).sub.4 (43 mg, 0.037 mmo). The mixture was stirred at rt for 1 h then concentrated in vacuo. The residue was purified by flash chromatography on silica gel (DCM/acetone: 100/0 to 0/100) to provide a mixture of expected intermediate and triphenylphosphine oxide. The mixture was dissolved in pyridine (0.55 mL) and sulfur trioxide pyridine complex (131 mg, 0.823 mmol) was added. The mixture was stirred at rt overnight then concentrated in vacuo. DCM (2 mL) was added to the residue and the precipitate filtered. The filtrate was concentrated and the residue purified by flash chromatography on silica gel (DCM/acetone: 50/50 to 0/100). The fractions containing the expected intermediate were combined and concentrated in vacuo. The residue was dissolved in a mixture of H.sub.2O/THF 7/3 (0.5 mL), and converted after ion exchange (Dowex sodium form column) to sodium [(2S,5R)-2-[[bis(tert-butoxycarbonylamino)methyleneamino]methyl]-7-oxo-3-thiazol-2-yl-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl] sulfate (39) (12 mg, 0.020 mmol, 28%).

    [0386] MS m/z ([M+H].sup.+) 575.

    [0387] MS m/z ([MH].sup.) 573.

    Step 3: Preparation of [(2S,5R)-2-(guanidinomethyl)-7-oxo-3-thiazol-2-yl-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl] hydrogen sulfate (Example 7)

    [0388] A solution of sodium [(2S,5R)-2-[[bis(tert-butoxycarbonylamino)methyleneamino]methyl]-7-oxo-3-thiazol-2-yl-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl] sulfate (39) (12 mg, 0.020 mmol) in anhydrous DCM (0.25 mL) was added to a mixture of DCM (0.45 mL) and TFA (0.45 mL) at 00 C. The mixture was stirred at 0 C. for 30 min and allowed to reach rt for 1 h. TFA (0.3 mL) was added and the mixture was stirred at rt for a further 3 h, then concentrated in vacuo. The residue was co-evaporated three times with DCM (2 mL). Water (0.5 mL) was added and the precipitate was filtered to provide [(2S,5R)-2-(guanidinommethyl)-7-oxo-3-thiazol-2-yl-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl] hydrogen sulfate (Example 7) (3.2 mg, 0.006 mmol, 42%) as a light pink solid.

    [0389] MS m/z ([M+H].sup.+) 375.

    [0390] MS m/z ([MH].sup.) 373.

    [0391] .sup.1H NMR (300 MHz, DMSO) 3.21-3.31 (m, 1H), 3.43-3.58 (m, 2H), 3.68-3.80 (m, 1H), 4.28-4.36 (m, 1H), 4.39 (dd, J=5.3, 2.4 Hz, 1H), 7.21 (d, J=5.3 Hz, 1H), 7.79 (d, J=3.2 Hz, 1H), 7.75-7.82 (m, 2H).

    Example 8: synthesis of sodium and 2,2,2-trifluoroacetate disalt of [(2S,5R)-2-[(3-aminopropanoylamino)methyl]-7-oxo-3-thiazol-2-yl-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl]sulfate

    [0392] ##STR00037##

    Step 1: Preparation of Intermediate (2S,5R)-6-allyloxy-2-(aminomethyl)-3-thiazol-2-yl-1,6-diazabicyclo[3.2.1]oct-3-en-7-one (40)

    [0393] A mixture of compound (20) (253 mg, 0.68 mmol) and NaN.sub.3 (221 mg, 3.41 mmol) in anhydrous DMF (3.0 mL) was stirred at 65 C. for 24 h. The mixture was poured in H.sub.2O (5 mL) and extracted with EtOAc (25 mL). The organic layer was washed with brine (5 mL), dried over Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was dissolved in anhydrous THF (2 mL) and anhydrous toluene (2 mL) and cooled at 0 C. under nitrogen atmosphere. A PMe.sub.3 solution 1M in THF (1.02 mL, 1.02 mmol) was dropwise added, the mixture was stirred at rt for 1 h and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (DCM/iPrOH: 100/0 to 50/50) to provide (2S,5R)-6-allyloxy-2-(aminomethyl)-3-thiazol-2-yl-1,6-diazabicyclo[3.2.1]oct-3-en-7-one (40) (100 mg, 0.34 mmol, 50%).

    [0394] MS m/z ([M+H].sup.+) 293.

    [0395] .sup.1H NMR (400 MHz, CDCl.sub.3) 2.81 (dd, J=14.1, 10.3 Hz, 1H), 3.31 (s, 2H), 3.44 (dd, J=14.1, 3.5 Hz, 1H), 3.98-4.04 (m, 1H), 4.37-4.49 (m, 3H), 5.27-5.32 (m, 2H), 5.33-5.39 (m, 1H), 5.95-6.08 (m, 1H), 6.93 (d, J=5.2 Hz, 1H), 7.24 (d, J=3.3 Hz, 1H), 7.75 (d, J=3.3 Hz, 1H).

    Step 2: Preparation of Intermediate tert-butyl N-[3-[[(2S,5R)-6-allyloxy-7-oxo-3-thiazol-2-yl-1,6-diazabicyclo[3.2.1]oct-3-en-2-yl]methylamino]-3-oxo-propyl]carbamate (41)

    [0396] To a solution of N-Boc--alanine (78 mg, 0.410 mmol) in anhydrous DMF (2 mL) at 0 C. under inert atmosphere was added HOBt hydrate (63 mg, 0.410 mmol), EDCI (72 mg, 0.376 mmol) and DIPEA (0.12 mL, 0.684 mmol). The mixture was stirred for 30 min at this temperature and a solution of compound (40) (100 mg, 0.342 mmol) in anhydrous DMF (2 mL) was added. The reaction mixture is allowed to reach rt and stirred for 1 hour. H.sub.2O was added and the mixture was extracted with EtOAc. The organic layer was washed with brine (5 mL), dried over Na.sub.2SO.sub.4, filtered and evaporated in vacuo. The crude was purified by flash chromatography on silica gel (DCM/EtOAc 100/0 to 0/100) to provide tert-butyl N-[3-[[(2S,5R)-6-allyloxy-7-oxo-3-thiazol-2-yl-1,6-diazabicyclo[3.2.1]oct-3-en-2-yl]methylamino]-3-oxo-propyl]carbamate (41) (95 mg, 0.205 mmol, 60%).

    [0397] MS m/z ([M+H].sup.+) 464.

    [0398] .sup.1H NMR (400 MHz, CDCl.sub.3) 1.43 (s, 9H), 2.42 (t, J=6.0 Hz, 2H), 5.30 (dd, J=11.3, 1.5 Hz, 1H), 3.36 (d, J=11.3 Hz, 1H), 3.39-3.48 (m, 3H), 4.04 (dd, J=5.3, 2.4 Hz, 1H), 4.10-4.20 (m, 1H), 4.36-4.49 (m, 2H), 4.62 (ddd, J=11.0, 4.5, 1.4 Hz, 1H), 5.25 (bs, 1H), 5.31 (dd, J=10.3, 1.4 Hz, 1H), 5.36 (dd, J=17.0, 1.4 Hz, 1H), 5.96-6.06 (m, 1H), 6.17 (bs, 1H), 6.98 (d, J=5.2 Hz, 1H), 7.24 (d, J=3.3 Hz, 1H), 7.75 (d, J=3.3 Hz, 1H).

    Step 3: Preparation of Intermediate sodium [(2S,5R)-2-[[3-(tert-butoxycarbonylamino)propanoylamino]methyl]-7-oxo-3-thiazol-2-yl-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl] sulfate (42)

    [0399] To a solution of compound (41) (85 mg, 0.183 mmol) in anhydrous DCM (1.85 mL) under nitrogen atmosphere were successively added AcOH (21 L, 0.367 mmol) and Pd(PPh.sub.3).sub.4 (106 mg, 0.091 mmol). After stirring for 2 h, dry pyridine (1.85 mL) and sulfur trioxide pyridine complex (145 mg, 0.915 mmol) were added and the resulting mixture was stirred for 2 hours. The reaction mixture was concentrated in vacuo, diluted with DCM (2 mL) and filtered. The filtrate was concentrated in vacuo. The crude was purified by flash chromatography on silica gel (DCM/acetone: 100/0 to 0/100). The fractions containing the expected intermediate were combined and concentrated in vacuo. The residue was dissolved in H.sub.2O (1 mL) and converted after ion exchange (Dowex sodium form column), and a chromatography on reverse phase C-18 (water/ACN: 100/0 to 0/100) to sodium [(2S,5R)-2-[[3-(tert-butoxycarbonylamino)propanoylamino]methyl]-7-oxo-3-thiazol-2-yl-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl] sulfate (42) (20 mg, 0.038 mmol, 21%) as a white solid.

    [0400] MS m/z ([MH].sup.) 502.

    [0401] .sup.1H NMR (400 MHz, D.sub.2O) 1.41 (s, 9H), 2.42 (t, J=6.4 Hz, 2H), 3.29-3.40 (m, 2H), 3.45-3.53 (m, 1H), 3.57-3.68 (m, 2H), 3.75 (dd, J=14.6, 4.5 Hz, 1H), 4.51 (dd, J=5.3, 2.5 Hz, 1H), 4.60 (ddd, J=10.4, 4.5, 1.4 Hz, 1H), 7.11 (d, J=5.3 Hz, 1H), 7.57 (d, J=3.3 Hz, 1H), 7.82 (d, J=3.3 Hz, 1H).

    Step 4: Preparation of Intermediate sodium and 2,2,2-trifluoroacetate disalt of [(2S,5R)-2-[(3-aminopropanoylamino)methyl]-7-oxo-3-thiazol-2-yl-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl] sulfate (example 8)

    [0402] Using the procedure described in example 2 (step 14) the intermediate (42) (17 mg, 0.032 mmol) is converted into Example 8 (16.1 mg, 0.030 mmol, 93%).

    [0403] MS m/z ([M+H].sup.+) 404.

    [0404] MS m/z ([MH].sup.) 402.

    [0405] .sup.1H NMR (300 MHz, D.sub.2O) 2.69 (d, J=6.7 Hz, 2H), 3.26 (d, J=6.7 Hz, 2H), 3.44-3.67 (m, 3H), 3.72 (dd, J=14.7, 4.1 Hz, 1H), 4.50-4.58 (m, 2H), 7.14-7.19 (m, 1H), 7.66 (d, J=3.5 Hz, 1H), 7.87 (d, J=3.5 Hz, 1H).

    Example 9: Biological Activity

    Method 1: -lactamase inhibitory activity, determination of IC.SUB.50 .(table 1)

    [0406] 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), 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).

    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 0.0032 0.0013 0.0054 0.0078 0.0041 0.0099 0.037 0.080 0.0041 0.00065 0.00051 Example 2 0.0017 0.00074 0.00076 0.0039 0.0032 0.017 0.11 0.0061 0.0068 0.00084 0.0035 Example 3 0.0030 0.0046 0.0020 0.033 0.0074 0.025 0.22 0.017 0.048 0.0016 0.0057 Example 4 0.014 0.020 0.0030 0.068 0.035 0.012 0.40 0.061 0.16 0.0073 0.027 Example 5 0.11 0.064 0.13 1.8 11 12 12 0.37 0.43 0.0080 0.18 Example 6 0.28 0.34 0.51 5.1 6.4 6.8 38 1.2 1.6 0.068 0.84

    Method 2: MIC of Compounds and Synerqy with Ceftazidime Against Bacterial Isolates (Tables 2-4)

    [0407] 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 (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 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.

    TABLE-US-00002 TABLE 2 Bacterial species used in MIC determination Strain 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 Derepressed 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

    TABLE-US-00003 TABLE 3 MIC of compounds MIC compounds of the invention alone (g/mL) Strains Example 1 Example 2 Example 3 Example 4 Example 5 Example 6 Example 7 260508 0.063 0.016 8 UFR61O 0.031 0.031 0.5 BAA-1898 >32 16 >32 >32 16 >32 >32 160143 >32 8 32 UFR68 >32 16 32 P99 0.031 0.016 0.063 >32 0.016 0.125 0.031 UFR85 >32 16 8 UFR70 16 4 8 UFR77 >32 32 >32 UFR74 >32 16 16 UFR18 0.031 0.016 0.016 131119 32 0.016 0.5 >32 0.063 >32 0.031 UFR21 >32 16 32 UFR24 >32 >32 >32 6299 >32 >32 >32 >32 >32 >32 >32 RGN238 >32 8 >32 >32 16 >32 16 200047 >32 16 16 190317 0.031 0.016 0.25 >32 0.016 0.125 0.063 UFR32 0.031 0.016 0.063 UFR38 0.016 32 0.016 210102 >32 4 4 UFR52 2 4 4 CIP107051 2 2 16 >32 2 16 8 CIP105250 >32 8 >32 >32 4 32 32 UFR35 1 2 2 0.25 UFR90 0.5 1 1 0.25 UFR92 2 2 8 1 UFR93 8 4 16 4 UFR47 2 2 4 1 UFR48 4 2 8 2 UFR59 8 8 16 2

    TABLE-US-00004 TABLE 4 MIC of Ceftazidime/compound combinations combination of CAZ and compounds of the invention et 4 g/mL: MIC (g/mL) Strains CAZ Example 1 Example 2 Example 3 Example 4 Example 5 Example 6 Example 7 260508 128 0.125 <0.25 <0.25 UFR610 128 <0.25 <0.25 <0.25 BAA-1898 256 0.5 0.125 1 64 0.125 4 0.125 160143 128 1 0.125 <0.25 UFR68 >128 1 0.125 0.25 P99 128 <0.25 <0.25 <0.125 64 <0.25 0.125 <0.125 UFR85 128 2 0.125 <0.25 UFR70 >128 1 <0.25 <0.25 UFR77 64 2 2 <0.25 UFR74 64 4 0.125 <0.25 UFR18 >128 <0.25 <0.25 <0.25 131119 0.5 <0.25 <0.25 <0.25 UFR21 128 0.5 0.125 <0.25 UFR24 >128 4 0.25 <0.25 6299 256 0.5 4 1 128 8 4 2 RGN238 0.5 0.125 0.125 <0.25 200047 128 0.5 0.125 <0.25 190317 128 <0.25 <0.25 <0.125 4 <0.25 <0.125 <0.125 UFR32 >128 <0.25 <0.25 <0.25 UFR38 >128 <0.25 <0.25 <0.25 210102 >128 0.5 0.125 <0.25 UFR52 >128 <0.25 <0.25 <0.25 CIP107051 256 <0.25 <0.25 0.25 64 <0.25 32 1 CIP105250 256 0.125 0.125 8 128 <0.25 64 32 UFR35 2 <0.25 <0.25 <0.25 <0.25 UFR90 64 <0.25 <0.25 <0.25 <0.25 UFR92 32 <0.25 <0.25 0.25 <0.25 UFR93 >128 <0.25 <0.25 0.5 <0.25 UFR47 >128 <0.25 <0.25 <0.25 <0.25 UFR48 256 <0.25 <0.25 16 <0.25 UFR59 128 <0.25 <0.25 2 <0.25