(12) United States Patent (10) Patent No.: US 8,822.450 B2 Patel Et Al

USOO88224.5OB2

(12) United States Patent (10) Patent No.: US 8,822.450 B2 Patel et al. (45) Date of Patent: Sep. 2, 2014

(54) 1,6-DIAZABICYCLO 3.2.1. OCTAN-7-ONE A6II 45/06 (2006.01) DERVATIVES AND THEIR USE IN THE A613 L/407 (2006.01) TREATMENT OF BACTERAL INFECTIONS A613 L/496 (2006.01) A63/67 (2006.01) (75) Inventors: Mahesh Vithalbhai Patel, Aurangabad A613 L/427 (2006.01) (IN); Prasad Keshav Deshpande, (52) U.S. Cl. Aurangabad (IN); Satish Bhawsar, USPC ...... 514/210.2: 514/210.05; 514/264.1; Aurangabad (IN); Sachin Bhagwat, 514/252.16; 514/210.18; 514/233.2: 514/210.16; Aurangabad (IN); Mohammed Alam 514/224.2: 514/253.08: 514/282; 514/119 Jafri. Aligarh (IN); Amit Mishra, (58) Field of Classification Search Aurangabad (IN); Laxmikant Pavase, USPC ...... 514/210.05, 264. 1, 252.16, 210.18, Aurangabad (IN); Sunil Gupta, Kota 514/233.2, 210.16, 224.2, 210.2, 253.08: (IN); Rajesh Kale, Aurangabad (IN); 544/282, 119 Sanjeev Joshi, Aurangabad (IN) See application file for complete search history. (73) Assignee: Wockhardt Ltd., Bandra-Kurla (56) References Cited Complex, Bandra East, Mumbai (IN) U.S. PATENT DOCUMENTS (*) Notice: Subject to any disclaimer, the term of this 2010/0093784 A1 4/2010 Ledoussal et al. patent is extended or adjusted under 35 U.S.C. 154(b) by 0 days. FOREIGN PATENT DOCUMENTS (21) Appl. No.: 14/111,592 WO 2009/09 1856 * 7/2009 WO WO 2009091856 A2 T 2009 (22) PCT Filed: Aug. 24, 2012 * cited by examiner (86). PCT No.: PCT/B2O12/O54290 Primary Examiner — Niloofar Rahmani S371 (c)(1) (74) Attorney, Agent, or Firm — Bio Intellectual Property

(2),s (4) Date: Dec. 20,9 2013 Services LLC (BioIPS); O. (Sam) Zaghmout (57) ABSTRACT (87) PCT Pub. No.: WO2013/030733 Compounds of Formula (I), their preparation and use in pre PCT Pub. Date: Mar. 7, 2013 venting or treating bacterial infection is disclosed. (65) Prior Publication Data US 2014/0148431A1 May 29, 2014 O Formula (I) (30) Foreign Application Priority Data R1 NN Aug. 27, 2011 (IN) ...... 2412AMUMA2011 H N

(51) Int.CO7D Cl. 47/08 (2006.01) O ul no R A6 IK3I/542 (2006.01) A 6LX3/529 (2006.01) A6 IK3I/429 (2006.01) 14 Claims, No Drawings US 8,822,450 B2 1. 1,6-DIAZABICYCLO 3,2,1 OCTAN-7-ONE wherein: DERVATIVES AND THEIR USE IN THE R is: TREATMENT OF BACTERAL INFECTIONS (a) hydrogen, (b) (CO), R. RELATED PATENT APPLICATIONS (c) COOR or (d) COCHCOR This application claims the benefit of Indian Provisional n is 0, 1 or 2; Patent Application No. 2412/MUM/2011 filed on Aug. 27, R is: 2011, the disclosures of which are incorporated herein by (a) SOM, reference in its entirety as if fully rewritten herein. All refer 10 (b) SONH, ences including patents, patent applications, and literature (c) POM, cited in the specification are expressly incorporated herein by (d) CHCOOM, reference in their entirety. (e) CFCOOM, 15 (f) CHFCOOM, or FIELD OF THE INVENTION (g) CF; M is hydrogen or a cation; The invention relates to nitrogen containing compounds, R is: their preparation and their use in preventing and/or treating (a) hydrogen, bacterial infections. (b) C-C alkyl optionally substituted with one or more Substituents independently selected from halogen, BACKGROUND OF THE INVENTION ORs, CN, COORs, CONRR, NRR, NRCORs, NRCONRR-7, heterocyclyl, heteroaryl, cycloalkyl Emergence of bacterial resistance to known antibacterial or aryl, agents is becoming a major challenge in treating bacterial 25 infections. One way forward to treat bacterial infections, and (c) CN, especially those caused by resistant bacteria, is to develop (d) NRR, newer antibacterial agents that can overcome the bacterial (e) CONRR, (f) NHCONRR, resistance. Coates et al. (Br. J. Pharmacol. 2007: 152(8), (g) aryl optionally substituted with one or more Substitu 1147-1154.) have reviewed novel approaches to developing 30 new antibiotics. However, the development of new antibac ents independently selected from C-C alkyl, ORs, terial agents is a challenging task. For example, Gwynn et al. NRR, halogen, CN, CONRR-7, SO-alkyl, SO (Annals of the New York Academy of Sciences, 2010, 1213: aryl, OSO-alkyl, OSO-aryl, or NHCONRR, 5-19) have reviewed the challenges in the discovery of anti (h) heterocyclyl optionally substituted with one or more bacterial agents. 35 Substituents independently selected from C-C alkyl, Several antibacterial agents have been described in the ORs, NRR, halogen, CN, CONRR, SO-alkyl, prior art (for example, see PCT International Application SO-aryl, OSO-alkyl, OSO-aryl, NHC(NH) Nos. PCT/US2010/060923, PCT/EP2010/067647, PCT/ NRR, or NHCONRR, US2010/052109, PCT/US2010/048109, PCT/GB2009/ (i) heteroaryl optionally substituted with one or more 050609, PCT/EP2009/0561.78 and PCT/US2009/041200). 40 Substituents independently selected from C-C alkyl, However, there remains a need for potent antibacterial agents ORs, NRR-7, halogen, CN, CONRR-7, SO-alkyl, for preventing and/or treating bacterial infections, including SO-aryl, OSO-alkyl, OSO-aryl, or NHCONRR, those caused by bacteria that are resistant to known antibac (j) cycloalkyl optionally substituted with one or more terial agents. Substituents independently selected from C-C alkyl, The inventors have Surprisingly discovered nitrogen con 45 ORs, NRR-7, halogen, CN, CONRR-7, SO-alkyl, taining compounds with antibacterial properties. SO-aryl, OSO-alkyl, OSO-aryl, or NHCONRR, (k) cycloalkyl substituted with C-C alkyl wherein SUMMARY OF THE INVENTION C-C alkyl is further substituted with one or more Accordingly there are provided nitrogen containing com substituents independently selected from ORs, 50 NRR, halogen, CN, or CONRR, or pounds, methods for preparation of these compounds, phar (1) ORs: maceutical compositions comprising these compounds, and R is: method for preventing or treating bacterial infection in a (a) hydrogen, Subject using these compounds. (b) C-C alkyl optionally substituted with one or more In one general aspect, there are provided compounds of 55 Substituents independently selected from halogen, Formula (I): OR, CN, COOR, CONRR, NRR, NRCORs, heterocyclyl, heteroaryl, cycloalkyl or aryl, Formula (I) (c) aryl optionally substituted with one or more substitu ents independently selected from C-C alkyl, ORs, 60 NRR, halogen, CN, CONRR-7, SO-alkyl, SO aryl, OSO-alkyl, OSO-aryl, or NHCONRR, (d) heterocyclyl optionally substituted with one or more Substituents independently selected from C-C alkyl, ORs, NRR-7, halogen, CN, CONRR-7, SO-alkyl, 65 SO-aryl, OSO-alkyl, OSO-aryl, or NHCONRR, or a stereoisomer or a pharmaceutically acceptable salt (e) heteroaryl optionally substituted with one or more thereof Substituents independently selected from C-C alkyl, US 8,822,450 B2 3 4 ORs, NRR-7, halogen, CN, CONRR-7, SO-alkyl, In another general aspect, there are provided pharmaceu SO-aryl, OSO-alkyl, OSO-aryl, or NHCONRR, tical compositions comprising: (a) a compound of Formula O (I), or a stereoisomer or a pharmaceutically acceptable salt (f) cycloalkyl optionally substituted with one or more thereof, and (b) at least one beta-lactamase inhibitor selected Substituents independently selected from C-C alkyl, from Sulbactam, taZobactam, clavulanic acid, or a pharma ORs, NRR-7, halogen, CN, CONRR-7, SO-alkyl, ceutically acceptable derivative thereof. SO-aryl, OSO-alkyl, OSO-aryl, or NHCONRR: In another general aspect, there are provided pharmaceu Rs and Rs are each independently: tical compositions comprising: (a) a compound of Formula (a) hydrogen, or (I), or a stereoisomer or a pharmaceutically acceptable salt (b) C-C alkyl optionally substituted with one or more 10 thereof, and (b) at least one antibacterial agent or a pharma Substituents independently selected from halogen, ceutically acceptable derivative thereof. CN, CONRR-7, NRR-7, heterocyclyl, heteroaryl, In another general aspect, there are provided pharmaceu cycloalkyl or aryl; tical compositions comprising: (a) a compound of Formula R and R, are each independently: (I), or a stereoisomer or a pharmaceutically acceptable salt (a) hydrogen, 15 thereof, (b) at least one beta-lactamase inhibitor selected from (b) C-C alkyl optionally substituted with one or more Sulbactam, taZobactam, clavulanic acid, or a pharmaceuti Substituents independently selected from halogen, cally acceptable derivative thereof, and (c) at least one anti ORs, CN, COORs, CONRRs, NRRs, NRCORs, bacterial agent or a pharmaceutically acceptable derivative heterocyclyl, heteroaryl, cycloalkyl or aryl, thereof. (c) aryl optionally substituted with one or more substitu In another general aspect, there is provided a method for ents independently selected from C-C alkyl, ORs, preventing or treating bacterial infection in a Subject, said NRRs halogen, CN, CONRRs, SO-alkyl, SO method comprising administering to said subject a pharma aryl, OSO-alkyl, OSO-aryl, or NHCONRRs, ceutically effective amount of a pharmaceutical composition (d) heterocyclyl optionally substituted with one or more comprising: (a) a compound of Formula (I), or a stereoisomer Substituents independently selected from C-C alkyl, 25 or a pharmaceutically acceptable salt thereof, and (b) at least ORs, NRSRs, halogen, CN, CONRRs, SO-alkyl, one beta-lactamase inhibitor selected from Sulbactam, taZo SO-aryl, OSO-alkyl, OSO-aryl, or NHCONRRs, bactam, clavulanic acid, or a pharmaceutically acceptable (e) heteroaryl optionally substituted with one or more derivative thereof. Substituents independently selected from C-C alkyl, In yet another general aspect, there is provided a method ORs, NRRs halogen, CN, CONRRs, SO-alkyl, 30 for preventing or treating a bacterial infection in a Subject, SO-aryl, OSO-alkyl, OSO-aryl, or NHCONRRs, said infection being caused by bacteria producing one or (f) cycloalkyl optionally substituted with one or more more beta-lactamase enzymes, wherein the method com Substituents independently selected from C-C alkyl, prises administering to said Subject a pharmaceutically effec ORs, NRSRs, halogen, CN, CONRRs, SO-alkyl, tive amount of a pharmaceutical composition comprising: (a) SO-aryl, OSO-alkyl, OSO-aryl, or NHCONRRs, 35 a compound of Formula (I), or a stereoisomer or a pharma O ceutically acceptable salt thereof, and (b) at least one beta (g) Re and R, are joined together to form a four to seven lactamase inhibitor selected from Sulbactam, taZobactam, member ring. clavulanic acid, or a pharmaceutically acceptable derivative In another general aspect, there are provided pharmaceu thereof. tical compositions comprising a compound of Formula (I), or 40 In another general aspect, there is provided a method for a stereoisomer or a pharmaceutically acceptable Salt thereof. preventing or treating bacterial infection in a Subject, said In another general aspect, there is provided a method for method comprising administering to said subject a pharma preventing or treating bacterial infection in a Subject, said ceutically effective amount of a pharmaceutical composition method comprising administering to said Subject a pharma comprising: (a) a compound of Formula (I), or a stereoisomer ceutically effective amount of a compound of Formula (I) or 45 or a pharmaceutically acceptable salt thereof, and (b) at least a stereoisomer or a pharmaceutically acceptable Salt thereof. one antibacterial agent or a pharmaceutically acceptable In another general aspect, there is provided a method for derivative thereof. preventing or treating a bacterial infection in a subject, said In yet another general aspect, there is provided a method infection being caused by bacteria producing one or more for preventing or treating a bacterial infection in a Subject, beta-lactamase enzymes, wherein the method comprises 50 said infection being caused by bacteria producing one or administering to said Subject a pharmaceutically effective more beta-lactamase enzymes, wherein the method com amount of a compound of Formula (I) or a stereoisomer or a prises administering to said Subject a pharmaceutically effec pharmaceutically acceptable salt thereof. tive amount of a pharmaceutical composition comprising: (a) In another general aspect, there is provided a method for a compound of Formula (I), or a stereoisomer or a pharma preventing or treating bacterial infection in a Subject, said 55 ceutically acceptable salt thereof, and (b) at least one antibac method comprising administering to said Subject a pharma terial agent or a pharmaceutically acceptable derivative ceutically effective amount of a pharmaceutical composition thereof. comprising a compound of Formula (I) or a stereoisomer or a In another general aspect, there is provided a method for pharmaceutically acceptable salt thereof. preventing or treating bacterial infection in a Subject, said In yet another general aspect, there is provided a method 60 method comprising administering to said subject a pharma for preventing or treating a bacterial infection in a subject, ceutically effective amount of a pharmaceutical composition said infection being caused by bacteria producing one or comprising: (a) a compound of Formula (I), or a stereoisomer more beta-lactamase enzymes, wherein the method com or a pharmaceutically acceptable salt thereof, (b) at least one prises administering to said Subject a pharmaceutically effec beta-lactamase inhibitor selected from Sulbactam, taZobac tive amount of a pharmaceutical composition comprising a 65 tam, clavulanic acid, or a pharmaceutically acceptable deriva compound of Formula (I) or a stereoisomer or a pharmaceu tive thereof, and (c) at least one antibacterial agent or a phar tically acceptable salt thereof. maceutically acceptable derivative thereof. US 8,822,450 B2 5 6 In yet another general aspect, there is provided a method In another general aspect, there are provided methods for for preventing or treating a bacterial infection in a subject, increasing antibacterial effectiveness of a antibacterial agent said infection being caused by bacteria producing one or in a Subject, said method comprising co-administering said more beta-lactamase enzymes, wherein the method com antibacterial agent or a pharmaceutically acceptable deriva prises administering to said Subject a pharmaceutically effec tive thereof with a pharmaceutically effective amount of a tive amount of a pharmaceutical composition comprising: (a) compound of Formula (I) or a stereoisomer or a pharmaceu a compound of Formula (I), or a stereoisomer or a pharma tically acceptable salt thereof. ceutically acceptable salt thereof, (b) at least one beta-lacta The details of one or more embodiments of the invention mase inhibitor selected from Sulbactam, taZobactam, clavu are set forth in the description below. Other features, objects lanic acid, or a pharmaceutically acceptable derivative 10 thereof, and (c) at least one antibacterial agent or a pharma and advantages of the invention will be apparent from the ceutically acceptable derivative thereof. following description including claims. In another general aspect, there is provided a method for preventing or treating bacterial infection in a Subject, said DETAILED DESCRIPTION OF THE INVENTION method comprising administering to said Subject a pharma 15 ceutically effective amount of: (a) a compound of Formula Reference will now be made to the exemplary embodi (I), or a stereoisomer or a pharmaceutically acceptable salt ments, and specific language will be used herein to describe thereof, and (b) at least one beta-lactamase inhibitor selected the same. It should nevertheless be understood that no limi from Sulbactam, taZobactam, clavulanic acid, or a pharma tation of the scope of the invention is thereby intended. Alter ceutically acceptable derivative thereof. ations and further modifications of the inventive features In yet another general aspect, there is provided a method illustrated herein, and additional applications of the prin for preventing or treating a bacterial infection in a subject, ciples of the invention as illustrated herein, which would said infection being caused by bacteria producing one or occur to one skilled in the relevant art and having possession more beta-lactamase enzymes, wherein the method com of this disclosure, are to be considered within the scope of the prises administering to said Subject a pharmaceutically effec 25 invention. It must be noted that, as used in this specification tive amount of: (a) a compound of Formula (I), or a stereoi and the appended claims, the singular forms “a” “an and Somer or a pharmaceutically acceptable salt thereof, and (b)at “the include plural referents unless the content clearly dic least one beta-lactamase inhibitor selected from Sulbactam, tates otherwise. All references including patents, patent appli taZobactam, clavulanic acid, or a pharmaceutically accept cations, and literature cited in the specification are expressly able derivative thereof. 30 incorporated herein by reference in their entirety. In another general aspect, there is provided a method for The inventors have Surprisingly discovered novel nitrogen preventing or treating bacterial infection in a subject, said containing compounds having antibacterial properties. method comprising administering to said Subject a pharma The term "C-C alkyl” as used herein refers to branched or ceutically effective amount of: (a) a compound of Formula unbranched acyclic hydrocarbon radical with 1 to 6 carbon (I), or a stereoisomer or a pharmaceutically acceptable salt 35 atoms. Typical, non-limiting examples of "C-C alkyl thereof, and (b) at least one antibacterial agent or a pharma include methyl, ethyl, n-propyl, iso-propyl. n-butyl, iso-bu ceutically acceptable derivative thereof. tyl, tert-butyl, n-pentyl, iso-pentyl, n-hexyl and the like. The In yet another general aspect, there is provided a method "C-C alkyl may be unsubstituted, or substituted with one for preventing or treating a bacterial infection in a subject, or more Substituents. Typical, non-limiting examples of Such said infection being caused by bacteria producing one or 40 substituents include halogen, alkoxy, CN, COOH, CONH, more beta-lactamase enzymes, wherein the method com OH, -NH2, —NHCOCH cycloalkyl, heterocyclyl, het prises administering to said Subject a pharmaceutically effec eroaryl, aryland the like. tive amount of: (a) a compound of Formula (I), or a stereoi The term “cycloalkyl as used herein refers to three to Somer or a pharmaceutically acceptable salt thereof, and (b)at seven member cyclic hydrocarbon radicals. The cycloalkyl least one antibacterial agent or a pharmaceutically acceptable 45 group optionally incorporates one or more double or triple derivative thereof. bonds, or a combination of double bonds and triple bonds, but In another general aspect, there is provided a method for which is not aromatic. Typical, non-limiting examples of preventing or treating bacterial infection in a Subject, said cycloalkyl groups include cyclopropane, cyclobutane, cyclo method comprising administering to said Subject a pharma pentane, cyclohexane, and cycloheptane. The cycloalkyl may ceutically effective amount of: (a) a compound of Formula 50 be unsubstituted, or substituted with one or more substituents. (I), or a stereoisomer or a pharmaceutically acceptable salt Typical, non-limiting examples of Such Substituents include thereof, (b) at least one beta-lactamase inhibitor selected from C-C alkyl, halogen, alkoxy, CN, COOH, CONH, OH, Sulbactam, taZobactam, clavulanic acid, or a pharmaceuti NH, NHCOCH, heterocyclyl, heteroaryl, aryl, SO-alkyl, cally acceptable derivative thereof, and (c) at least one anti SO-aryl, OSO-alkyl, —OSO-aryland the like. bacterial agent or a pharmaceutically acceptable derivative 55 The term "heterocyclyl as used herein refers to four to thereof. seven member cycloalkyl group containing one or more het In yet another general aspect, there is provided a method eroatoms selected from nitrogen, oxygen or Sulfur. The het for preventing or treating a bacterial infection in a subject, erocycloalkyl group optionally incorporates one or more said infection being caused by bacteria producing one or double or triple bonds, or a combination of double bonds and more beta-lactamase enzymes, wherein the method com 60 triple bonds, but which is not aromatic. Typical, non-limiting prises administering to said Subject a pharmaceutically effec examples of heterocycloalkyl groups include aZetidine, pyr tive amount of: (a) a compound of Formula (I), or a stereoi rolidine, 2-oxo-pyrrolidine, imidazolidin-2-one, piperidine, Somer or a pharmaceutically acceptable salt thereof, (b) at oxazine, thiazine, piperazine, piperazin-2,3-dione, morpho least one beta-lactamase inhibitor selected from Sulbactam, line, thiamorpholine, azapane, and the like. The heterocy taZobactam, clavulanic acid, or a pharmaceutically accept 65 cloalkyl may be unsubstituted, or substituted with one or able derivative thereof, and (c) at least one antibacterial agent more Substituents. Typical, non-limiting examples of Such or a pharmaceutically acceptable derivative thereof. Substituents include C-C alkyl, halogen, alkoxy, CN, US 8,822,450 B2 7 8 COOH, CONH, OH, NH, NHCOCH, heterocyclyl, het therefore, can form acidic salts (formed with inorganic and/or eroaryl, aryl, SO-alkyl, SO-aryl, OSO-alkyl, OSO-aryl organic acids), as well as basic salts (formed with inorganic and the like. and/or organic bases). Such salts can be prepared using pro The term “aryl as used herein refers to a monocyclic or cedures described in the art. For example, the basic moiety polycyclic aromatic hydrocarbon. Typical, non-limiting can be converted to its salt by treating a compound with a examples of aryl groups include phenyl, naphthyl, anthrace Suitable amount of acid. Typical, non-limiting examples of nyl, fluorenyl, phenanthrenyl, and the like. The aryl group such suitable acids include hydrochloric acid, trifluoroacetic may be unsubstituted, or substituted with one or more sub acid, methanesulphonic acid, or the like. Alternatively, the stituents. Typical, non-limiting examples of such Substituents acid moiety may be converted into its salt by treating with a include C-C alkyl, halogen, alkoxy, CN, COOH, CONH2, 10 Suitable base. Typical non-limiting examples of Such bases OH, NH, NHCOCH, heterocyclyl, heteroaryl, aryl, SO include Sodium carbonate, sodium bicarbonate, potassium alkyl, SO-aryl, OSO-alkyl, OSO-aryl and the like. carbonate, potassium bicarbonate or the like. In case of com The term "heteroarylas used herein refers to a monocyclic pounds containing more than one functional groups capable or polycyclic aromatic hydrocarbon group wherein one or of being converted into salt, each Such functional group may more carbon atoms have been replaced with heteroatoms 15 be converted to salt independently. For example, in case of selected from nitrogen, oxygen, and Sulfur. If the heteroaryl compounds containing two basic nitrogen atoms, one basic group contains more than one heteroatom, the heteroatoms nitrogen can form salt with one acid while the other basic may be the same or different. Typical, non-limiting example nitrogen can form salt with another acid. Some compounds of heteroaryl groups include 1,2,4-oxadiaZol. 1,3,4-oxadia according to the invention contain both, acidic as well as basic Zol. 1,3,4-thiadiazol. 1.2.3,4-tetraZol, 1.3-oxazol. 1.3-thiaz moieties, and thus can form inner salts or corresponding ole, pyridine, pyrimidine, pyrazine, pyridazine, furan, pyrrol, Zwitterions. In general, all pharmaceutically acceptable salt thiophene, imidazole, pyrazole, benzofuran, benzothiophene, forms of compounds of Formula (I) according to invention benzimidazole, benzoxazole, benzothiazole, thiazole, and the including acid addition salts, base addition salts, Zwitterions like. The heteroaryl group may be unsubstituted, or substi or the like are contemplated to be within the scope of the tuted with one or more substituents. Typical, non-limiting 25 present invention and are generically referred to as pharma examples of Such Substituents include C-C alkyl, halogen, ceutically acceptable salts. alkoxy, CN, COOH, CONH, OH, NH, NHCOCH, hetero The term “halogen' or “halo' as used herein refers to cyclyl, heteroaryl, aryl, SO-alkyl, SO-aryl, OSO-alkyl, chlorine, bromine, fluorine, or iodine. OSO-aryland the like. The term “infection' or “bacterial infection' as used herein The term “stereoisomers' as used herein refers to com 30 includes presence of bacteria, in or on a Subject, which, if its pounds that have identical chemical constitution, but differ growth were inhibited, would resultina benefit to the subject. with regard to the arrangement of their atoms or groups in As such, the term “infection” in addition to referring to the space. The compounds of Formula (I) may contain asymmet presence of bacteria also refers to normal flora, which is not ric or chiral centers and, therefore, exist in different stereoi desirable. The term “infection' includes infection caused by Someric forms. It is intended, unless specified otherwise, that 35 bacteria. all stereoisomeric forms of the compounds of Formula (I) as The term “treat”, “treating or “treatment as used herein well as mixtures thereof, including racemic mixtures, form refers to administering a medicament, including a pharma part of the present invention. In addition, all geometric and ceutical composition, or one or more pharmaceutically active positional isomers (including cis and trans-forms) as well as ingredients, for prophylactic and/or therapeutic purposes. mixtures thereof, are also embraced within the scope of the 40 The term “prophylactic treatment” refers to treating a subject invention. In general, a reference to a compound is intended to who is not yet infected, but who is susceptible to, or otherwise cover its stereoisomers and mixture of various stereoisomers. at a risk of infection (preventing the bacterial infection). The The term “optionally substituted as used herein means term “therapeutic treatment” refers to administering treat that substitution is optional and therefore includes both ment to a subject already suffering from infection. The terms unsubstituted and substituted atoms and moieties. A “substi 45 “treat”, “treating or “treatment as used herein also refer to tuted atom or moiety indicates that any hydrogen on the administering compositions or one or more of pharmaceuti designated atom or moiety can be replaced with a selection cally active ingredients discussed herein, with or without from the indicated substituent group, provided that the nor additional pharmaceutically active or inert ingredients, in mal valency of the designated atom or moiety is not exceeded, order to: (i) reduce or eliminate either a bacterial infection or and that the Substitution results in a stable compound. 50 one or more symptoms of the bacterial infection, or (ii) retard The term “pharmaceutically acceptable salt as used herein the progression of a bacterial infection or of one or more refers to one or more salts of a given compound which pos symptoms of the bacterial infection, or (iii) reduce the sever sesses the desired pharmacological activity of the free com ity of a bacterial infection or of one or more symptoms of the pound and which are neither biologically nor otherwise unde bacterial infection, or (iv) suppress the clinical manifestation sirable. In general, the “pharmaceutically acceptable salts' 55 of a bacterial infection, or (v) Suppress the manifestation of refer to salts that are suitable for use in contact with the tissues adverse symptoms of the bacterial infection. of human and animals without undue toxicity, irritation, aller The term “pharmaceutically effective amount’ or “thera gic response and the like, and are commensurate with a rea peutically effective amount’ or “effective amount’ as used sonable benefit/risk ratio. Pharmaceutically acceptable salts herein refers to an amount, which has a therapeutic effect or is are well known in the art. For example, S. M. Berge, et al. (J. 60 the amount required to produce a therapeutic effect in a Sub Pharmaceutical Sciences, 66: 1-19 (1977)), incorporated ject. For example, a therapeutically or pharmaceutically herein by reference in its entirety, describes various pharma effective amount of an antibacterial agent or a pharmaceutical ceutically acceptable salts in details. composition is the amount of the antibacterial agent or the In general, the compounds according to the invention con pharmaceutical composition required to produce a desired tain basic (e.g. nitrogen atoms) as well as acid moieties (e.g. 65 therapeutic effect as may be judged by clinical trial results, compounds of Formula (I) wherein M is hydrogen). A person model animal infection studies, and/or in vitro studies (e.g. in of skills in the art would appreciate that such compounds, agar or broth media). The pharmaceutically effective amount US 8,822,450 B2 10 depends on several factors, including but not limited to, the enzymes that are produced by bacteria and have the ability to microorganism (e.g. bacteria) involved, characteristics of the hydrolyze the beta-lactam ring in a beta-lactam compound, Subject (for example height, weight, sex, age and medical either partially or completely. history), severity of infection and the particular type of the The term “beta-lactamase inhibitor” as used herein refers antibacterial agent used. For prophylactic treatments, a thera 5 to a compound capable of inhibiting activity of one or more peutically or prophylactically effective amount is that amount beta-lactamase enzymes, either partially or completely. which would be effective in preventing a microbial (e.g. bac The term “pharmaceutically inert ingredient’ or “carrier' terial) infection. or “excipient” refers to a compound or material used to facili The term “administration” or “administering includes tate administration of a compound, including for example, to delivery of a composition or one or more pharmaceutically 10 increase the solubility of the compound. Typical, non-limit active ingredients to a Subject, including for example, by any ing examples of Solid carriers include, starch, lactose, dical appropriate methods, which serves to deliver the composition cium phosphate. Sucrose, and kaolin and so on. Typical, non or its active ingredients or other pharmaceutically active limiting examples of liquid carriers include, sterile water, ingredients to the site of the infection. The method of admin saline, buffers, non-ionic Surfactants, and edible oils such as istration may vary depending on various factors, such as for 15 oil, peanut and sesame oils and so on. In addition, various example, the components of the pharmaceutical composition adjuvants commonly used in the art may be included. These or the nature of the pharmaceutically active or inert ingredi and other such compounds are described in the literature, for ents, the site of the potential or actual infection, the microor example, in the Merck Index (Merck & Company, Rahway, ganism involved, severity of the infection, age and physical N.J.). Considerations for inclusion of various components in condition of the Subject and a like. Some non-limiting pharmaceutical compositions are described, for example, in examples of ways to administer a composition or a pharma Gilman et al. (Eds.) (1990); Goodman and Gilmans: The ceutically active ingredient to a Subject according to this Pharmacological Basis of Therapeutics, 8th Ed., Pergamon invention includes oral, intravenous, topical, intrarespiratory, Press., which is incorporated herein by reference in its intraperitoneal, intramuscular, parenteral, Sublingual, trans entirety. dermal, intranasal, aerosol, intraocular, intratracheal, 25 The term “subject' as used herein refers to vertebrate or intrarectal, vaginal, gene gun, dermal patch, eye drop, ear invertebrate, including a mammal. The term “subject’ drop or mouthwash. In case of a pharmaceutical composition includes human, animal, a bird, a fish, or an amphibian. comprising more than one ingredient (active or inert), one of Typical, non-limiting examples of a “subject' includes way of administering such composition is by admixing the humans, cats, dogs, horses, sheep, bovine cows, pigs, lambs, ingredients (e.g. in the form of a suitable unit dosage form 30 rats, mice and guinea pigs. Such as tablet, capsule, Solution, powder and a like) and then The term “pharmaceutically acceptable derivative' as used administering the dosage form. Alternatively, the ingredients herein refers to and includes any pharmaceutically acceptable may also be administered separately (simultaneously or one salt, pro-drugs, metabolites, esters, ethers, hydrates, poly after the other) as long as these ingredients reach beneficial morphs, Solvates, complexes, enantiomers or adducts of a therapeutic levels such that the composition as a whole pro 35 compound described herein which, upon administration to a vides a synergistic and/or desired effect. subject, is capable of providing (directly or indirectly) the The term “growth' as used herein refers to a growth of one parent compound. For example, the term “antibacterial agent or more microorganisms and includes reproduction or popu or a pharmaceutically acceptable derivative thereof includes lation expansion of the microorganism (e.g. bacteria). The all derivatives of the antibacterial agent (such as salt, pro term also includes maintenance of on-going metabolic pro 40 drugs, metabolites, esters, ethers, hydrates, polymorphs, Sol cesses of a microorganism, including processes that keep the Vates, complexes, enantiomers or adducts) which, upon microorganism alive. administration to a subject, is capable of providing (directly The term, “effectiveness” as used herein refers to ability of or indirectly) the antibacterial compound. a treatment or a composition or one or more pharmaceutically In general, the term “cation' includes Na, K, Mg, Ca, active ingredients to produce a desired biological effect in a 45 NH", (CHCH)N" etc. subject. For example, the term “antibacterial effectiveness” In one general aspect, there are provided compounds of of a composition oran antibacterial agent refers to the ability of the composition or the antibacterial agent to prevent or Formula (I): treat the microbial (e.g. bacterial) infection in a Subject. The term “synergistic' or “synergy” as used herein refers 50 to the interaction of two or more agents so that their combined Formula (I) effect is greater than their individual effects. The term “antibacterial agent” as used herein refers to any Substance, compound or a combination of Substances or a combination compounds capable of: (i) inhibiting, reducing 55 or preventing growth of bacteria; (ii) inhibiting or reducing ability of a bacteria to produce infection in a subject; or (iii) inhibiting or reducing ability of bacteria to multiply or remain infective in the environment. The term “antibacterial agent' or a stereoisomer or a pharmaceutically acceptable salt also refers to compounds capable of decreasing infectivity or 60 thereof; virulence of bacteria. wherein: The term “beta-lactam antibacterial agent” as used herein R is: refers to compounds with antibacterial properties and con (a) hydrogen, taining a beta-lactam nucleus in their molecular structure. (b) (CO), R. The term “beta-lactamase' as used herein refers to any 65 (c) COOR or enzyme or protein or any other Substance that breaks down a (d) COCHCOR beta-lactam ring. The term “beta-lactamase' includes n is 0, 1 or 2; US 8,822,450 B2 11 12 R is: Rs and Rs are each independently: (a) SOM, (a) hydrogen, or (b) SONH (b) C-C alkyl optionally substituted with one or more (c) POM, substituents independently selected from halogen, CN, (d) CHCOOM, CONRR, NRR, heterocyclyl, heteroaryl, cycloalkyl (e) CFCOOM, or aryl; (f) CHFCOOM, or Re and R, are each independently: (g) CF, (a) hydrogen, M is hydrogen or a cation; (b) C-C alkyl optionally substituted with one or more R is: 10 Substituents independently selected from halogen, ORs, (a) hydrogen, CN, COOR, CONRRs. NRRs, NRCORs, heterocy (b) C-C alkyl optionally substituted with one or more clyl, heteroaryl, cycloalkyl or aryl, Substituents independently selected from halogen, ORs, (c) aryl optionally substituted with one or more substitu CN, COOR, CONRR, NRR, NRCORs, ents independently selected from C-C alkyl, ORs, NRCONRR-7, heterocyclyl, heteroaryl, cycloalkyl or 15 NRRs halogen, CN, CONRRs, SO-alkyl, SO-aryl, aryl, OSO-alkyl, OSO-aryl, or NHCONRRs, (c) CN, (d) heterocyclyl optionally substituted with one or more (d) NRR-7, Substituents independently selected from C-C alkyl, (e) CONRR, ORs, NRSRs, halogen, CN, CONRRs, SO-alkyl, SO (f) NHCONRR, aryl, OSO-alkyl, OSO-aryl, or NHCONRRs, (g) aryl optionally Substituted with one or more substitu (e) heteroaryl optionally substituted with one or more sub ents independently selected from C-C alkyl, ORs, stituents independently selected from C-C alkyl, ORs, NRR, halogen, CN, CONRR, SO-alkyl, SO-aryl, NRRs halogen, CN, CONRRs, SO-alkyl, SO-aryl, OSO-alkyl, OSO-aryl, or NHCONRR, 25 OSO-alkyl, OSO-aryl, or NHCONRRs, (h) heterocyclyl optionally substituted with one or more (f) cycloalkyl optionally substituted with one or more sub Substituents independently selected from C-C alkyl, stituents independently selected from C-C alkyl, ORs, ORs, NRR-7, halogen, CN, CONRR-7, SO-alkyl, SO NRRs, halogen, CN, CONRRs, SO-alkyl, SO-aryl, aryl, OSO-alkyl, OSO-aryl, NHC(NH)NRR, or OSO-alkyl, OSO-aryl, or NHCONRRs, or 30 (g) R and R, are joined together to form a four to seven NHCONRR, member ring. (i) heteroaryl optionally substituted with one or more sub Typical non-limiting examples of compounds according to stituents independently selected from C-C alkyl, ORs, the invention include: NRR, halogen, CN, CONRR-7, SO-alkyl, SO-aryl, trans-sulfuric acid mono-2-(N' (S)-pyrrolidin-2-carbo OSO-alkyl, OSO-aryl, or NHCONRR, 35 nyl-hydrazinocarbonyl)-7-oxo-1,6-diaza-bicyclo[3.2.1]oct (j) cycloalkyl optionally substituted with one or more sub 6-ylester; stituents independently selected from C-C alkyl, ORs, trans-sulfuric acid mono-2-(N' ((R)-piperidin-3-carbo NRR, halogen, CN, CONRR-7, SO-alkyl, SO-aryl, nyl)-hydrazinocarbonyl)-7-oxo-1,6-diaza-bicyclo[3.2.1]oct OSO-alkyl, OSO-aryl, or NHCONRR, 6-ylester; (k) cycloalkyl substituted with C-C alkyl wherein C-C, 40 trans-sulfuric acid mono-2-(N' (R)-pyrrolidin-3-car alkyl is further substituted with one or more substituents bonyl-hydrazinocarbonyl)-7-oxo-1,6-diaza-bicyclo3.2.1 independently selected from ORs, NRR, halogen, CN, oct-6-ylester; or CONRR, or trans-7-oxo-2-N'-((R)-piperidin-3-carbonyl)-hydrazi (1) ORs: nocarbonyl-1,6-diaza-bicyclo[3.2.1-oct-6-yloxy)-acetic R is: 45 acid; (a) hydrogen, trans-difluoro-7-oxo-2-N'-((R)-piperidin-3-carbonyl)- (b) C-C alkyl optionally substituted with one or more hydrazinocarbonyl-1,6-diaza-bicyclo[3.2.1-oct-6-yloxy}- Substituents independently selected from halogen, ORs, acetic acid; CN, COORs, CONRR, NRR, NRCORs, heterocy trans-Sulfuric acid mono-2-hydrazinocarbonyl-7-oxo-1, clyl, heteroaryl, cycloalkyl or aryl, 50 6-diaza-bicyclo3.2.1]oct-6-ylester; trans-Sulfuric acid mono-2-(N'-(amino-acetyl)-hydrazi (c) aryl optionally substituted with one or more substitu nocarbonyl)-7-oxo-1,6-diaza-bicyclo[3.2.1]oct-6-ylester; ents independently selected from C-C alkyl, ORs, trans-Sulfuric acid mono-2-(N'-(3-amino-propioyl)-hy NRR, halogen, CN, CONRR, SO-alkyl, SO-aryl, drazinocarbonyl)-7-oxo-1,6-diaza-bicyclo[3.2.1]oct-6-yl) OSO-alkyl, OSO-aryl, or NHCONRR, 55 ester, (d) heterocyclyl optionally substituted with one or more trans-Sulfuric acid mono-2-(N'-(4-amino-butanoyl)-hy Substituents independently selected from C-C alkyl, drazinocarbonyl)-7-oxo-1,6-diaza-bicyclo[3.2.1]oct-6-yl) ORs, NRR-7, halogen, CN, CONRR-7, SO-alkyl, SO ester, aryl, OSO-alkyl, OSO-aryl, or NHCONRR, trans-sulfuric acid mono-2-(N'-((2S)-2-amino-3-hy (e) heteroaryl optionally substituted with one or more sub 60 droxy-propioyl)-hydrazinocarbonyl)-7-OXO-1,6-diaza-bicy stituents independently selected from C-C alkyl, ORs, clo3.2.1]oct-6-ylester; NRR, halogen, CN, CONRR-7, SO-alkyl, SO-aryl, trans-sulfuric acid mono-2-(N'-(2S,4S)-4-fluoro-pyrroli OSO-alkyl, OSO-aryl, or NHCONRR, or din-2-carbonyl-hydrazinocarbonyl)-7-OXO-1,6-diaza-bicy (f) cycloalkyl optionally substituted with one or more sub clo3.2.1]oct-6-ylester; stituents independently selected from C-C alkyl, ORs, 65 trans-sulfuric acid mono-2-(N'-(2S,4R)-4-methoxy-pyr NRR, halogen, CN, CONRR-7, SO-alkyl, SO-aryl, rolidin-2-carbonyl-hydrazinocarbonyl)-7-oxo-1,6-diaza-bi OSO-alkyl, OSO-aryl, or NHCONRR: cyclo[3.2.1]oct-6-ylester;

US 8,822,450 B2 15 described in PCT International Publication No. WO 2009/ -continued 091856, was reacted with corresponding acid hydrazides in O presence of a suitable coupling agent such as EDC hydrochlo ride, or dicyclohexylcarbodiimide (DCC), in a solvent such as Ris-sly N.N dimethyl formamide; N.N dimethylacetamide: 1.4 diox NisriH ane; chloroform, dichloromethane; or dichloroethane at a O N temperature ranging from -15°C. to 60° C. for about 1 to 24 hours to obtain intermediate compound (1b). N The intermediate compound (1b) was subjected for hydro O OSOR genolysis in presence of a suitable catalyst (e.g. 5% or 10% 10 R = Hor Na palladium on carbon, or 20% palladium hydroxide on carbon) Compound of invention in presence of a suitable hydrogen source (such as hydrogen of Formula-1 gas, ammonium formate, cyclohexene) in a Suitable solvent (such as methanol, ethanol, methanol-dichloromethane mix ture, or N.N dimethyl formamide-dichloromethane mixture) 15 The intermediate compound (1c) was sulfonated by react at a temperature ranging from about 25°C. to 60°C. for about ing it with a Sulfonating reagent (Such as Sulfur trioxide 1 to 14 hours to obtain intermediate compound (1c). pyridine complex, or sulfur trioxide-N,N-dimethyl forma mide complex) in a suitable solvent (such as pyridine, N.N- dimethyl formamide) at a temperature ranging from about Scheme-1 25°C. to 90° C. for about 1 to 24 hours to obtain pyridine salt of sulfonic acid which when treated with tetrabutyl ammo nium sulfate provided terabutylammonium salt of sulfonic acid as an intermediate compound (1 d). 25 Some compounds according to the invention were isolated N RCONHNH He as a Zwitterions, by treating intermediate compound (1d) with Coupling agent trifluoroacetic acid, in a Suitable solvent (such as dichlo romethane, chloroform, oracetonitrile) at a temperature rang ing from about -10° C. to 40° C. for about 1 to 14 hours, 30 especially when R in intermediate compound (1 d) contained tert-butoxycarbonyl protected amine function. la O Some other compounds according to the invention were isolated as a corresponding sodium salt, by passing interme Risusly 35 diate compound (1 d) through sodium form of Aberlite 200C H resin in a tetrahydrofuran-water mixture followed by evapo O N Hydro ration of the solvent under vacuum. geno N n lysis O He O 40 Scheme-2 O

1b Risusly o, O H 45 rsrs R1 f, y O N -- NN H Sulfonating agent V N -e- Jr. r O OH then BuNHSO4. 50 1c A. NV O OH A. A O Base Br N-1 -

55 O A = H or F Sodium salt formation O -e- R1 lyA, O OSONBu Nish--O N i) Hydrolysis -e- A. A ii) Deprotec N / N O tion (if R1 CFCOOH O O N1 bearsa (when R mine func Boc-aminocontainst 65 O tion with group) 2d Boc group) US 8,822,450 B2 17 18 -continued Sulbactam, taZobactam, clavulanic acid, or a pharmaceuti O cally acceptable derivative thereof. Ris-sly In some other embodiments, there are provided pharma NN , t ceutical compositions comprising: (a) a compound of For H mula (I), or a stereoisomer or a pharmaceutically acceptable O N A. A salt thereof, and (b) at least one antibacterial agent or a phar N maceutically acceptable derivative thereof. V OH In some other embodiments, there are provided pharma O O ceutical compositions comprising: (a) a compound of For O 10 mula (I), or a stereoisomer or a pharmaceutically acceptable Compound of invention salt thereof, (b) at least one beta-lactamase inhibitor selected of formula-1 from Sulbactam, taZobactam, clavulanic acid, or a pharma ceutically acceptable derivative thereof, and (c) at least one antibacterial agent or a pharmaceutically acceptable deriva As described in Scheme-2, the hydroxyl intermediate (1c) 15 tive thereof. obtained as per Scheme-1, was subjected for alkylation with In some other embodiments, there is provided a method for alkylating agent (such as ethyl-bromoacetate, ethyl-fluoroac preventing or treating bacterial infection in a Subject, said etate or ethyl-difluoroacetate) in presence of a base (Such as method comprising administering to said subject a pharma potassium carbonate, diisopropylethylamine or triethy ceutically effective amount of a pharmaceutical composition lamine) in a suitable solvent (such as N.N dimethyl forma comprising: (a) a compound of Formula (I), or a stereoisomer mide, N.N dimethyl acetamide or N-methylpyrrolidine) to or a pharmaceutically acceptable salt thereof, and (b) at least provide O-alkylated compound (2d). one beta-lactamase inhibitor selected from Sulbactam, taZo The compound (2d) was subjected for hydrolysis in pres bactam, clavulanic acid, or a pharmaceutically acceptable ence of a base (such as lithium hydroxide or potassium derivative thereof. 25 In some other embodiments, there is provided a method for hydroxide) in a Suitable solvent (such as aqueous tetrahydro preventing or treating a bacterial infection in a subject, said furan, aquous dioxane) to provide compound of Formula (I) infection being caused by bacteria producing one or more after pH adjustment. beta-lactamase enzymes, wherein the method comprises Optionally, if R bears amine function protected with Boc administering to said Subject a pharmaceutically effective group, then it was removed in an additional step of deprotec 30 amount of a pharmaceutical composition comprising: (a) a tion by using a Suitable deprotecting agent (such as trifluoro compound of Formula (I), or a stereoisomer or a pharmaceu acetic acid or HF pyridine) in a solvent (such as dichlo tically acceptable salt thereof, and (b) at least one beta-lacta romethane, chloroform or acetonitrile) to provide compound mase inhibitor selected from Sulbactam, taZobactam, clavu of Formula (I). lanic acid, or a pharmaceutically acceptable derivative In some embodiments, there are provided pharmaceutical 35 thereof. compositions comprising a compound of Formula (I), or a In some other embodiments, there is provided a method for Stereoisomer or a pharmaceutically acceptable salt thereof. preventing or treating bacterial infection in a Subject, said In some other embodiments, there is provided a method for method comprising administering to said subject a pharma preventing or treating bacterial infection in a Subject, said ceutically effective amount of a pharmaceutical composition method comprising administering to said Subject a pharma 40 comprising: (a) a compound of Formula (I), or a stereoisomer ceutically effective amount of a compound of Formula (I) or or a pharmaceutically acceptable salt thereof, and (b) at least a stereoisomer or a pharmaceutically acceptable Salt thereof. one antibacterial agent or a pharmaceutically acceptable In some embodiments, there is provided a method for pre derivative thereof. venting or treating a bacterial infection in a subject, said In some other embodiments, there is provided a method for infection being caused by bacteria producing one or more 45 preventing or treating a bacterial infection in a subject, said beta-lactamase enzymes, wherein the method comprises infection being caused by bacteria producing one or more administering to said Subject a pharmaceutically effective beta-lactamase enzymes, wherein the method comprises amount of a compound of Formula (I) or a stereoisomer or a administering to said Subject a pharmaceutically effective pharmaceutically acceptable salt thereof. amount of a pharmaceutical composition comprising: (a) a In some other embodiments, there is provided a method for 50 compound of Formula (I), or a stereoisomer or a pharmaceu preventing or treating bacterial infection in a Subject, said tically acceptable salt thereof, and (b) at least one antibacte method comprising administering to said Subject a pharma rial agent or a pharmaceutically acceptable derivative thereof. ceutically effective amount of a pharmaceutical composition In some other embodiments, there is provided a method for comprising a compound of Formula (I) or a stereoisomer or a preventing or treating bacterial infection in a Subject, said pharmaceutically acceptable salt thereof. 55 method comprising administering to said subject a pharma In some other embodiments, there is provided a method for ceutically effective amount of a pharmaceutical composition preventing or treating a bacterial infection in a subject, said comprising: (a) a compound of Formula (I), or a stereoisomer infection being caused by bacteria producing one or more or a pharmaceutically acceptable salt thereof, (b) at least one beta-lactamase enzymes, wherein the method comprises beta-lactamase inhibitor selected from Sulbactam, taZobac administering to said Subject a pharmaceutically effective 60 tam, clavulanic acid, or a pharmaceutically acceptable deriva amount of a pharmaceutical composition comprising a com tive thereof, and (c) at least one antibacterial agent or a phar pound of Formula (I) or a stereoisomer or a pharmaceutically maceutically acceptable derivative thereof. acceptable salt thereof. In some other embodiments, there is provided a method for In some embodiments, there are provided pharmaceutical preventing or treating a bacterial infection in a subject, said compositions comprising: (a) a compound of Formula (I), or 65 infection being caused by bacteria producing one or more a stereoisomer or a pharmaceutically acceptable Salt thereof, beta-lactamase enzymes, wherein the method comprises and (b) at least one beta-lactamase inhibitor selected from administering to said Subject a pharmaceutically effective US 8,822,450 B2 19 20 amount of a pharmaceutical composition comprising: (a) a pharmaceutically acceptable Salt thereof, and (b) Sulbactam compound of Formula (I), or a stereoisomer or a pharmaceu or a pharmaceutically acceptable derivative thereof. tically acceptable salt thereof, (b) at least one beta-lactamase In some other embodiments, there is provided a method for inhibitor selected from Sulbactam, taZobactam, clavulanic preventing or treating a bacterial infection in a subject, said acid, or a pharmaceutically acceptable derivative thereof, and 5 method comprising administering to said subject a pharma (c) at least one antibacterial agent or a pharmaceutically ceutically effective amount of: (a) trans-Sulfuric acid mono acceptable derivative thereof. 2-(N' (R)-piperidin-3-carbonyl-hydrazinocarbonyl)-7- In some other embodiments, there is provided a method for oxo-1,6-diaza-bicyclo[3.2.1]oct-6-ylester or a stereoisomer preventing or treating bacterial infection in a Subject, said or a pharmaceutically acceptable salt thereof, and (b) Sulbac method comprising administering to said Subject a pharma 10 tam or a pharmaceutically acceptable derivative thereof. In some other embodiments, there is provided a method for ceutically effective amount of: (a) a compound of Formula preventing or treating a bacterial infection in a subject, said (I), or a stereoisomer or a pharmaceutically acceptable salt infection being caused by bacteria producing one or more thereof, and (b) at least one beta-lactamase inhibitor selected beta-lactamase enzymes, said method comprising adminis from Sulbactam, taZobactam, clavulanic acid, or a pharma 15 tering to said Subject a pharmaceutically effective amount of ceutically acceptable derivative thereof. (a) trans-sulfuric acid mono-2-(N' (R)-piperidin-3-carbo In some other embodiments, there is provided a method for nyl-hydrazinocarbonyl)-7-oxo-1,6-diaza-bicyclo[3.2.1]oct preventing or treating a bacterial infection in a subject, said 6-ylester or a stereoisomer or a pharmaceutically acceptable infection being caused by bacteria producing one or more salt thereof, and (b) Sulbactam or a pharmaceutically accept beta-lactamase enzymes, wherein the method comprises able derivative thereof. administering to said Subject a pharmaceutically effective In some other embodiments, there is provided a method for amount of: (a) a compound of Formula (I), or a stereoisomer preventing or treating bacterial infection in a Subject, said or a pharmaceutically acceptable salt thereof, and (b) at least method comprising administering to said subject a pharma one beta-lactamase inhibitor selected from Sulbactam, taZo ceutically effective amount of a pharmaceutical composition bactam, clavulanic acid, or a pharmaceutically acceptable 25 comprising: (a) trans-Sulfuric acid mono-2-(N'-(R)-pip derivative thereof. eridin-3-carbonyl-hydrazinocarbonyl)-7-oxo-1,6-diaza-bi In some other embodiments, there is provided a method for cyclo3.2.1]oct-6-ylester or a stereoisomer or a pharmaceu preventing or treating bacterial infection in a Subject, said tically acceptable salt thereof, and (b) sulbactam or a method comprising administering to said Subject a pharma pharmaceutically acceptable derivative thereof. ceutically effective amount of: (a) a compound of Formula 30 In some embodiments, there is provided a method for pre (I), or a stereoisomer or a pharmaceutically acceptable salt venting or treating a bacterial infection in a Subject, said thereof, and (b) at least one antibacterial agent or a pharma infection being caused by bacteria producing one or more ceutically acceptable derivative thereof. beta-lactamase enzymes, wherein the method comprises In some other embodiments, there is provided a method for administering to said Subject a pharmaceutically effective preventing or treating a bacterial infection in a subject, said 35 amount of a pharmaceutical composition comprising: (a) infection being caused by bacteria producing one or more trans-sulfuric acid mono-2-(N' (R)-piperidin-3-carbo beta-lactamase enzymes, wherein the method comprises nyl-hydrazinocarbonyl)-7-oxo-1,6-diaza-bicyclo[3.2.1]oct administering to said Subject a pharmaceutically effective 6-ylester or a stereoisomer or a pharmaceutically acceptable amount of: (a) a compound of Formula (I), or a stereoisomer salt thereof, and (b) Sulbactam or a pharmaceutically accept or a pharmaceutically acceptable salt thereof, and (b) at least 40 able derivative thereof. one antibacterial agent or a pharmaceutically acceptable In some embodiments, there are provided pharmaceutical derivative thereof. compositions comprising: (a) trans-Sulfuric acid mono-2- In some other embodiments, there is provided a method for (N' (R)-pyrrolidin-3-carbonyl-hydrazinocarbonyl)-7- preventing or treating bacterial infection in a Subject, said oxo-1,6-diaza-bicyclo[3.2.1]oct-6-ylester or a stereoisomer method comprising administering to said Subject a pharma 45 or a pharmaceutically acceptable salt thereof, and (b) Sulbac ceutically effective amount of: (a) a compound of Formula tam or a pharmaceutically acceptable derivative thereof. (I), or a stereoisomer or a pharmaceutically acceptable salt In some other embodiments, there is provided a method for thereof, (b) at least one beta-lactamase inhibitor selected from preventing or treating a bacterial infection in a subject, said Sulbactam, taZobactam, clavulanic acid, or a pharmaceuti method comprising administering to said subject a pharma cally acceptable derivative thereof, and (c) at least one anti 50 ceutically effective amount of: (a) trans-Sulfuric acid mono bacterial agent or a pharmaceutically acceptable derivative 2-(N' (R)-pyrrolidin-3-carbonyl-hydrazinocarbonyl)-7- thereof. oxo-1,6-diaza-bicyclo[3.2.1]oct-6-ylester or a stereoisomer In some other embodiments, there is provided a method for or a pharmaceutically acceptable salt thereof, and (b) Sulbac preventing or treating a bacterial infection in a subject, said tam or a pharmaceutically acceptable derivative thereof. infection being caused by bacteria producing one or more 55 In some other embodiments, there is provided a method for beta-lactamase enzymes, wherein the method comprises preventing or treating a bacterial infection in a subject, said administering to said Subject a pharmaceutically effective infection being caused by bacteria producing one or more amount of: (a) a compound of Formula (I), or a stereoisomer beta-lactamase enzymes, said method comprising adminis or a pharmaceutically acceptable salt thereof, (b) at least one tering to said Subject a pharmaceutically effective amount of beta-lactamase inhibitor selected from Sulbactam, taZobac 60 (a) trans-sulfuric acid mono-2-(N' (R)-pyrrolidin-3-car tam, clavulanic acid, or a pharmaceutically acceptable deriva bonyl-hydrazinocarbonyl)-7-oxo-1,6-diaza-bicyclo3.2.1 tive thereof, and (c) at least one antibacterial agent or a phar oct-6-ylester or a stereoisomer or a pharmaceutically accept maceutically acceptable derivative thereof. able salt thereof, and (b) sulbactam or a pharmaceutically In some embodiments, there are provided pharmaceutical acceptable derivative thereof. compositions comprising: (a) trans-Sulfuric acid mono-2- 65 In some other embodiments, there is provided a method for (N' (R)-piperidin-3-carbonyl-hydrazinocarbonyl)-7-oxo preventing or treating bacterial infection in a Subject, said 1,6-diaza-bicyclo[3.2.1]oct-6-ylester or a stereoisomer or a method comprising administering to said subject a pharma US 8,822,450 B2 21 22 ceutically effective amount of a pharmaceutical composition Typical, non-limiting examples of Macrollide antibacterial comprising: (a) trans-Sulfuric acid mono-2-(N'-(R)-pyrro agents include Azithromycin, Clarithromycin, Dirithromy lidin-3-carbonyl-hydrazinocarbonyl)-7-oxo-1,6-diaza-bi cin, Erythromycin, Roxithromycin, Troleandomycin, cyclo3.2.1]oct-6-ylester or a stereoisomer or a pharmaceu Telithromycin, Spectinomycin, Solithromycin and the like. tically acceptable salt thereof, and (b) sulbactam or a Typical, non-limiting examples of Monobactam antibacte pharmaceutically acceptable derivative thereof. rial agents include Aztreonam and the like. In some embodiments, there is provided a method for pre Typical, non-limiting examples of Nitrofuran antibacterial venting or treating a bacterial infection in a subject, said agents include Furazolidone, Nitrofurantoin and the like. infection being caused by bacteria producing one or more Typical, non-limiting examples of Penicillin antibacterial beta-lactamase enzymes, wherein the method comprises 10 agents include Amoxicillin, Ampicillin, AZlocillin, Carbeni administering to said Subject a pharmaceutically effective cillin, Cloxacillin, Dicloxacillin, Flucloxacillin, Mezlocillin, amount of a pharmaceutical composition comprising: (a) Methicillin, Nafcillin, Oxacillin, Penicillin G, Penicillin V, trans-sulfuric acid mono-2-(N'—(R)-pyrrolidin-3-carbo Piperacillin, Temocillin, Ticarcillin and the like. nyl-hydrazinocarbonyl)-7-oxo-1,6-diaza-bicyclo[3.2.1]oct Typical, non-limiting examples of Polypeptide antibacte 15 rial agents include Bacitracin, Colistin, Polymyxin B and the 6-ylester or a stereoisomer or a pharmaceutically acceptable like. salt thereof, and (b) Sulbactam or a pharmaceutically accept Typical, non-limiting examples of Quinolone antibacterial able derivative thereof. agents include Ciprofloxacin, Enoxacin, Gatifloxacin, Levo In some embodiments, there are provided methods for floxacin, Lomefloxacin, Moxifloxacin, Nalidixic acid, increasing antibacterial effectiveness of a antibacterial agent Levonadifloxacin, Norfloxacin, Ofloxacin, Trovafloxacin, in a Subject, said method comprising co-administering said Grepafloxacin, Sparfloxacin, Temafloxacin and the like. antibacterial agent or a pharmaceutically acceptable deriva Typical, non-limiting examples of Sulfonamide antibacte tive thereof with a pharmaceutically effective amount of a rial agents include Mafenide, Sulfonamidochrysoidine, Sul compound of Formula (I) or a stereoisomer or a pharmaceu facetamide, Sulfadiazine, Sulfamethizole, Sulfamethox tically acceptable salt thereof. 25 azole, Sulfasalazine, Sulfisoxazole, Trimethoprim and the In Some embodiments, the compositions and methods like. according to the invention use compounds of Formula (I) or a Typical, non-limiting examples of Tetracycline antibacte Stereoisomer or a pharmaceutically acceptable salt thereof in rial agents include Demeclocycline, Doxycycline, Minocy combination with at least one antibacterial agent or a phar cline, Oxytetracycline, Tetracycline, Tigecycline and the like. maceutically acceptable derivative thereof. A wide variety of 30 Typical, non-limiting examples of Oxazolidinone antibac antibacterial agents can be used. Typical, non-limiting terial agents include Tedizolid, Linezolid, Ranbezolid, Tor examples of antibacterial agents include one or more of anti eZolid, Radezolid etc. bacterial compounds generally classified as aminoglyco The pharmaceutical compositions according to the inven sides, Ansamycins, Carbacephems, Cephalosporins, Cepha tion may include one or more pharmaceutically acceptable mycins, Lincosamides, Lipopeptides, Macrollides, 35 carriers or excipients or the like, Typical, non-limiting Monobactams, Nitrofurans, Penicillins, Polypeptides, Qui examples of such carriers or excipient include mannitol, lac nolones, Sulfonamides, Tetracyclines, Oxazolidinone and the tose, starch, magnesium Stearate, Sodium saccharine, talcum, like. cellulose, Sodium crosscarmellose, glucose, gelatin, Sucrose, Typical, non-limiting examples of Aminoglycoside anti magnesium carbonate, wetting agents, emulsifying agents, bacterial agents include Amikacin, Gentamicin, Kanamycin, 40 solubilizing agents, pH buffering agents, lubricants, stabiliz Neomycin, Netilmicin, Tobramycin, Paromomycin, Arbeka ing agents, binding agents etc. cin, Streptomycin, Apramycin and the like. The pharmaceutical compositions according to this inven Typical, non-limiting examples of Ansamycin antibacte tion can exist in various forms. In some embodiments, the rial agents include Geldanamycin, Herbimycin and the like. pharmaceutical composition is in the form of a powder or a Typical, non-limiting examples of Carbacephem antibac 45 Solution. In some other embodiments, the pharmaceutical terial agents include Loracarbef and the like. compositions according to the invention are in the form of a Typical, non-limiting examples of Carbapenem antibacte powder that can be reconstituted by addition of a compatible rial agents include Ertapenem, Doripenem, Imipenem, Mero reconstitution diluent prior to parenteral administration. Non penem and the like. limiting example of such a compatible reconstitution diluent Typical, non-limiting examples of Cephalosporin and 50 includes water. Cephamycin antibacterial agents include Cefazolin, In some other embodiments, the pharmaceutical composi Cefacetrile, Cefadroxil, Cefalexin, Cefaloglycin, Cefalo tions according to the invention are in the form of a frozen nium, Cefaloridine, Cefalotin, Cefapirin, Cefatrizine, composition that can be diluted with a compatible diluent Cefazedone, Cefazaflur, Cefradine, Cefroxadine, Ceftezole, prior to parenteral administration. Cefaclor, Cefamandole, Cefninox, Cefonicid, Ceforanide, 55 In some other embodiments, the pharmaceutical composi Cefotiam, Cefprozil, Cefbuperazone, Cefuroxime, Cefu tions according to the invention are in the form ready to use Zonam, Cephamycin, Cefoxitin, Cefotetan, Cefinetazole, for parenteral administration. Carbacephem, Cefixime, Ceftazidime, Ceftriaxone, Cefcap In the methods according to the invention, the pharmaceu ene, Cefdaloxime, Cefdinir, Cefditoren, Cefetamet, tical composition and/or other pharmaceutically active ingre Cefinenoxime, Cefodizime, CefoperaZone, Cefotaxime, Cef 60 dients disclosed herein may be administered by any appro pimizole, Ce?piramide, Cefpodoxime, Cefsulodin, Cefteram, priate method, which serves to deliver the composition or its Ceftibuten, Ceftiolene, Ceftizoxime, Oxacephem, Cefepime, constituents or the active ingredients to the desired site. The Cefozopran, Ce?pirome, Cefauinome, Ceftobiprole, Ceftio method of administration can vary depending on various fac fur, Cefauinome, Cefovecin, CXA-101, Ceftaroline, Cefto tors. Such as for example, the components of the pharmaceu biprole etc. 65 tical composition and nature of the active ingredients, the site Typical, non-limiting examples of Lincosamide antibacte of the potential or actual infection, the microorganism (e.g. rial agents include Clindamycin, Lincomycin and the like. bacteria) involved, severity of infection, age and physical US 8,822,450 B2 23 24 condition of the Subject. Some non-limiting examples of tract infection, intraabdominal infections, respiratory tract administering the composition to a Subject according to this infections, pneumonia (nosocomial), bacteremia meningitis, invention include oral, intravenous, topical, intrarespiratory, Surgical, infections etc. intraperitoneal, intramuscular, parenteral, Sublingual, trans Surprisingly, the compounds, compositions and methods according to the invention are also effective in preventing or dermal, intranasal, aerosol, intraocular, intratracheal, treating bacterial infections that are caused by bacteria pro intrarectal, vaginal, gene gun, dermal patch, eye drop, ear ducing one or more beta-lactamase enzymes. The ability of drop or mouthwash. compositions and methods according to the present invention The compositions according to the invention can be for to treat such resistant bacteria with typical beta-lactam anti mulated into various dosage forms wherein the active ingre biotics represents a significant improvement in the art. dients and/or excipients may be present either together (e.g. 10 In general, the compounds of Formula (I) or a stereoisomer as an admixture) or as separate components. When the vari or pharmaceutically acceptable salt thereof according to ous ingredients in the composition are formulated as a mix invention are also useful in increasing antibacterial effective ture, such composition can be delivered by administering ness of a antibacterial agent in a Subject. The antibacterial Such a mixture. The composition or dosage form wherein the effectiveness of one or more antibacterial agents may 15 increased, for example, by co-administering said antibacte ingredients do not come as a mixture, but come as separate rial agent or a pharmaceutically acceptable derivative thereof components, such composition/dosage form may be admin with a pharmaceutically effective amount of a compound of istered in several ways. In one possible way, the ingredients Formula (I) or a stereoisomer or a pharmaceutically accept may be mixed in the desired proportions and the mixture is able salt thereof according to the invention. then administered as required. Alternatively, the components It will be readily apparent to one skilled in the art that or the ingredients (active or inert) may be separately admin varying Substitutions and modifications may be made to the istered (simultaneously or one after the other) in appropriate invention disclosed herein without departing from the scope proportion so as to achieve the same or equivalent therapeutic and spirit of the invention. For example, those skilled in the level or effect as would have been achieved by administration art will recognize that the invention may be practiced using a of the equivalent mixture. 25 variety of different compounds within the described generic Similarly, in the methods according to the invention, the descriptions. active ingredients disclosed herein may be administered to a Subject in several ways depending on the requirements. In EXAMPLES Some embodiments, the active ingredients are admixed in The following examples illustrate the embodiments of the appropriate amounts and then the admixture is administered 30 invention that are presently best known. However, it is to be to a subject. In some other embodiments, the active ingredi understood that the following are only exemplary or illustra ents are administered separately. Since the invention contem tive of the application of the principles of the present inven plates that the active ingredients agents may be administered tion. Numerous modifications and alternative compositions, separately, the invention further provides for combining sepa methods, and systems may be devised by those skilled in the rate pharmaceutical compositions in kit form. The kit may 35 art without departing from the spirit and scope of the present comprise one or more separate pharmaceutical compositions, invention. The appended claims are intended to cover Such each comprising one or more active ingredients. Each of Such modifications and arrangements. Thus, while the present separate compositions may be present in a separate container invention has been described above with particularity, the Such as a bottle, vial, Syringes, boxes, bags, and the like. following examples provide further detail in connection with Typically, the kit comprises directions for the administration 40 what are presently deemed to be the most practical and pre of the separate components. The kit form is particularly ferred embodiments of the invention. advantageous when the separate components are preferably administered in different dosage forms (e.g., oral and Example-1 parenteral) ore are administered at different dosage intervals. When the active ingredients are administered separately, they 45 trans-sulfuric acid mono-2-(N' (S)-pyrrolidin-2- may be administered simultaneously or sequentially. carbonyl-hydrazinocarbonyl)-7-oxo-1,6-diaza-bicy The pharmaceutical composition or the active ingredients clo3.2.1]oct-6-ylester according to the present invention may be formulated into a variety of dosage forms. Typical, non-limiting examples of dosage forms include solid, semi-solid, liquid and aerosol 50 dosage forms. Such as tablets, capsules, powders, Solutions, Suspensions, Suppositories, aerosols, granules, emulsions, syrups, elixirs and a like. In general, the pharmaceutical compositions and method N disclosed herein are useful in preventing or treating bacterial 55 infections. Advantageously, the compositions and methods disclosed herein are also effective in preventing or treating infections caused by bacteria that are considered be less or not Susceptible to one or more of known antibacterial agents or their known compositions. Some non-limiting examples of 60 Such bacteria known to have developed resistance to various Step-1: Preparation of trans-2-N'-(6-benzyloxy-7- antibacterial agents include Acinetobacter, E. coli, oXo-1,6-diaza-bicyclo3.2.1]octane-2-carbonyl)- Pseudomonas aeruginosa, Staphylococcus aureus, Entero hydrazinocarbonyl-(S)-pyrrolidin-1-carboxylic acid bacter, Klebsiella, Citrobacter and a like. Other non-limiting tert-butyl ester examples of infections that may be prevented or treated using 65 the compositions and/or methods of the invention include: To a clear solution of trans-6-benzyloxy-7-oxo-1,6-diaza skin and Soft tissue infections, febrile neutropenia, urinary bicyclo[3.2.1]octane-2-carboxylic acid (15gm, 0.054 mol) in US 8,822,450 B2 25 26 N,N-dimethyl formamide (150 ml), was added EDC hydro combined organic extract was washed with brine (50 ml) and chloride (15.57gm, 0.082 mol) followed by HOBt (11.0gm, dried on Sodium Sulfate and evaporated under vacuum to 0.082 mol) at about 25° C. to 35° C. under stirring. The provide solid, that was triturated with diethyl ether and fil reaction mixture was stirred for 15 minutes and a solution tered to provide white powder as a step-3 product (tetrabuty of (S) N-tert-butoxycarbonyl-pyrrolidin-2-carboxylic acid lammonium salt of trans-2-N'-(6-sulfooxy-7-oxo-1,6-diaza hydrazide (14.93 gm, 0.065 mol) dissolved in N,N-dimethyl bicyclo[3.2.1]octane-2-carbonyl)-hydrazinocarbonyl-(S)- formamide (75 ml), followed by N.N-di-isopropyl ethy pyrrolidin-1-carboxylic acid tetr-butyl ester), in 3.0 gm lamine (28.4 ml, 0.163 mol) were added. The reaction mix quantity (83% yield). ture was stirred at a temperature between 25°C. to 35°C. for Analysis: MS (ES-) C.H.N.O.S.N(CH) as a 16 hours. The reaction mixture was poured under stirring into 10 salt=476.0 (M-1) as a free sulfonic acid; 10% aqueous citric acid solution (2250 ml). The resulting H'NMR (CDC1)=9.13 (brs, 1H), 8.49 (brs, 1H), 4.35 (br mixture was extracted with diethyl ether (1000 mlx3). Com bined organic layer was washed with water (1000 ml) fol s, 2H), 3.98 (d. 1H), 3.24-3.50 (m. 10H), 3.13 (brid, 1H), 2.35 lowed by brine solution (500 ml) and dried over sodium (dd, 2H), 2.16 (brs, 2H), 1.91-2.01 (m, 4H), 1.61-1.70 (m, Sulfate. Concentration of organic layer under vacuum 15 10H), 1.40-1.48 (m, 17H), 0.98-1.02 (m, 12H). afforded the crude residue in 13 gm quantity. The residue was purified using silica gel column chromatography to provide Step-4: trans-sulfuric acid mono-2-(N' (S)-pyrro the product (trans-2-N'-(6-benzyloxy-7-oxo-1,6-diaza-bicy lidin-2-carbonyl-hydrazinocarbonyl)-7-oxo-1,6- clo3.2.1]octane-2-carbonyl)-hydrazinocarbonyl-(S)-pyrro diaza-bicyclo[3.2.1]oct-6-ylester lidin-1-carboxylic acid tert-butyl ester) in 6.3 gm quantity as a white powder. To the powder obtained in step-3 (3.0 gm, 4.17 mmol) was Analysis: MS (ES+) CHNO. 488.1 (M+1): added, a solution of trifluoroacetic acid (7 ml) in dichlo H'NMR (DMSO-d)=9.86 (brid, 1H), 9.75 (brid, 1H), romethane (7ml) slowly by syringe at -5°C. over a period of 7.34-7.44 (m, 5H), 4.92 (dd, 2H), 4.07-4.10 (m. 1H), 3.78 5 minutes. The mixture was maintained under stirring for 1 hr. 25 Solvents were removed below 40°C. under high vacuum to 3.82 (m. 1H), 3.68 (brd, 1H), 3.20-3.25 (m, 3H), 2.87 (brid, provide a residue, which was triturated with diethyl ether (50 1H), 1.62-2.10 (m, 8H), 1.34 (s.9H). mlx5) and each time diethyl ether was decanted. The obtained Step-2: Preparation of trans-2-N'-(6-hydroxy-7-oxo white solid was further triturated with acetonitrile (100 1,6-diaza-bicyclo[3.2.1]octane-2-carbonyl)-hydrazi mlx2). The resultant solid was stirred in dichloromethane nocarbonyl-(S)-pyrrolidin-1-carboxylic acid tert 30 (100 nil) and the suspension was filtered. The solid was dried butyl ester under vacuum to provide title compound of the invention (trans-sulfuric acid mono-2-(N'-(S)-pyrrolidin-2-carbo To a clear solution of step-1 product (3.0gm, 6.15 mmol) in nyl-hydrazinocarbonyl)-7-oxo-1,6-diaza-bicyclo[3.2.1]oct methanol (30 ml) was added 10% palladium on carbon (300 6-ylester) in 1.2 gm quantity (59% yield). mg). The Suspension was stirred under atmospheric hydrogen 35 Analysis: MS (ES-) CHNO.S=376.2 (M-1) as a free pressure at a temperature of about 30° C. for 2 hours. The Sulfonic acid; catalyst was filtered over a celite bed and catalyst-containing H'NMR (DMSO-d)=10.39 (brs, 1H), 10.15 (s, 1H), 8.96 bed was washed with additional methanol (10 ml) and dichlo (brs, 2H), 4.19 (t, 1H), 4.03 (brs, 1H), 3.86 (d. 1H), 3.16-3.25 romethane (10 ml). The filtrate was concentrated in vacuum (m, 3H), 3.02 (brd, 1H), 2.27-2.33 (m. 1H), 1.92-2.23 (m, to provide a white powder, which was triturated with diethyl 40 1H), 1.84-1.90 (m, 4H), 1.69-1.75 (m. 1H), 1.54-1.62 (m, ether to provide trans-2-N'-(6-hydroxy-7-oxo-1,6-diaza-bi 1H). cyclo[3.2.1]octane-2-carbonyl)-hydrazinocarbonyl-(S)- pyrrolidin-1-carboxylic acid tert-butyl ester as a white pow Example-2 der in 2.00gm quantity in 82% yield. Analysis: MS (ES") CHNO=398.0 (M+1): 45 trans-sulfuric acid mono-2-(N' (R)-piperidin-3- H'NMR (DMSO-d)=9.82 (d. 1H), 9.70-9.80 (m, 2H), carbonyl-hydrazinocarbonyl)-7-oxo-1,6-diaza-bicy 4.08-4.15 (m. 1H), 3.4.0-3.78 (m, 1H), 3.59 (brs, 1H), 3.17 clo3.2.1]oct-6-ylester 3.40 (m,3H), 2.97 (brid, 1H), 1.55-2.15 (m, 8H), 1.35 (s.9H).

Step-3: Preparation of tetrabutylammonium salt of 50 trans-2-N'-(6-sulfooxy-7-oxo-1,6-diaza-bicyclo 3.2.1]octane-2-carbonyl)-hydrazinocarbonyl-(S)- pyrrolidin-1-carboxylic acid tetr-butyl ester

The product obtained in step-2 (2.00 gm, 5.03 mmol) was 55 dissolved in pyridine (40 ml) and to the clear solution was added pyridine sulfur trioxide complex (4.03 gm, 25.18 mmol). The suspension was stirred at a temperature 25°C. to 35° C. for overnight. The suspension was filtered and the solids were washed with dichloromethane (25 mlx2). The 60 filtrate was evaporated under vacuum and the residue was Step-1: Preparation of trans-3-N'-(6-benzyloxy-7- stirred in 0.5 N aqueous potassium dihydrogen phosphate oXo-1,6-diaza-bicyclo3.2.1]octane-2-carbonyl)- solution (200 ml) for 0.5 hour. The solution was washed with hydrazinocarbonyl-(R)-piperidin-1-carboxylic acid ethyl acetate (100 mlx4) and layers were separated. To the tert-butyl ester aqueous layer was added tetrabutylammonium Sulphate (1.71 65 gm, 5.03 mmol) and stirred for four hours at about 25°C. The By using the procedure described in Step-1 of Example-1 mixture was extracted with dichloromethane (100 mlx2). The above, and by using trans-6-benzyloxy-7-oxo-1,6-diaza-bi US 8,822,450 B2 27 28 cyclo[3.2.1]octane-2-carboxylic acid (25 gm, 0.084 mol), 2.86-2.91 (m. 1H), 2.65-2.66 (m, 1H), 1.97-2.03 (m, 1H), N,N-dimethyl formamide (625 ml), EDC hydrochloride (24 1.57-1.88 (m, 7H). gm, 0.126 mol), HOBt (16.96 gm, 0.126 mol), (R) N-tert C-32.6°, (c. 0.5, water). butoxycarbonyl-piperidin-3-carboxylic acid hydrazide Example-3 (21.40 gm, 0.088 mol) to provide the title compound in 17.0 gm quantity, 41% yield as a white solid. trans-sulfuric acid mono-2-(N' (R)-pyrrolidin-3- Analysis: MS (ES+) CHNO=502.1 (M-1); carbonyl-hydrazinocarbonyl)-7-oxo-1,6-diaza-bicy H'NMR (CDC1)=8.40 (brs, 1H), 7.34-7.44 (m, 5H), 5.05 clo3.2.1]oct-6-ylester (d. 1H), 4.90 (d. 1H), 4.00 (brid, 1H), 3.82 (brs, 1H), 3.30 (br 10 s, 1H), 3.16-3.21 (m, 1H), 3.06 (brd, 1H), 2.42 (brs, 1H), 2.29-2.34 (m. 1H), 1.18-2.02 (m, 4H), 1.60-1.75 (m, 4H), 1.45-1.55 (m, 2H), 1.44 (s, 9H). Step-2: Preparation of trans-3-N'-(6-hydroxy-7-oxo 15 1,6-diaza-bicyclo[3.2.1]octane-2-carbonyl)-hydrazi nocarbonyl-(R)-piperidin-1-carboxylic acid tert butyl ester By using the procedure described in Step-2 of Example-1 above, and by using trans-3-N'-(6-benzyloxy-7-oxo-1,6- diaza-bicyclo3.2.1]octane-2-carbonyl)-hydrazinocarbo Step-1: Preparation of trans-3-N'-(6-benzyloxy-7- nyl-(R)-piperidin-1-carboxylic acid tert-butyl ester (16.5 oXo-1,6-diaza-bicyclo3.2.1]octane-2-carbonyl)- gm, 0.033 mol), methanol (170 ml) and 10% palladium on 25 hydrazinocarbonyl-(R)-pyrrolidin-1-carboxylic acid carbon (3.5 gm) to provide the title compound in 13.5gm. tert-butyl ester quantity as a pale pink Solid and it was used for the next reaction immediately. By using the procedure described in Step-1 of Example-1, Analysis: MS (ES+) C.H.N.O. 411.1 (M+1): 30 and by using trans-6-benzyloxy-7-oxo-1,6-diaza-bicyclo 3.2.1]octane-2-carboxylic acid (15.7gm, 0.057 mol), N.N- Step-3: Preparation of tetrabutylammonium salt of dimethyl formamide (390 ml), EDC hydrochloride (16.24 trans-3-N'-(6-sulfooxy-7-oxo-1,6-diaza-bicyclo gm, 0.085 mol), HOBt (11.48 gm, 0.085 mol), (R) N-tert 3.2.1]octane-2-carbonyl)-hydrazinocarbonyl-(R)- butoxycarbonyl-pyrrolidin-3-carboxylic acid hydrazide piperidin-1-carboxylic acid tert-butyl ester 35 (13.7gm, 0.06 mol) to provide the title compound in 11.94 gm quantity, 43% yield as a white solid. By using the procedure described in Step-3 of Example-1 Analysis: MS (ES+) CHNO. 488.2 (M+1): above, and by using trans-3-N'-(6-hydroxy-7-oxo-1,6- H'NMR (CDC1), DO exchange=7.30-7.39 (m, 5H), 4.85 diaza-bicyclo3.2.1]octane-2-carbonyl)-hydrazinocarbo 40 (s. 2H), 3.77 (d. 1H), 3.68 (bris, 1H), 3.39-3.41 (m. 1H), nyl-(R)-piperidin-1-carboxylic acid tert-butyl ester (13.5 3.17-3.26 (m,3H), 3.01 (d. 1H), 2.90-2.92 (m, 2H), 1.97-2.03 gm, 0.033 mol), pyridine (70 ml) and pyridine sulfur trioxide (m. 2H), 1.79-1.89 (m, 2H), 1.66-1.70 (m, 1H), 1.55-1.57 (m, complex (26.11 gm, 0.164 mol), 0.5 N aqueous potassium 1H), 1.32 (s, 9H). dihydrogen phosphate solution (400 ml) and tetrabutylam monium sulphate (9.74 gm, 0.033 mol) to provide the title 45 Step-2: Preparation of trans-3-N'-(6-hydroxy-7-oxo compound in 25gm quantity as a yellowish solid, in quanti 1,6-diaza-bicyclo[3.2.1]octane-2-carbonyl)-hydrazi tative yield. nocarbonyl-(R)-pyrrolidin-1-carboxylic acid tert Analysis: MS (ES-) CHNO.S.N(CHo) as a butyl ester salt=490.0 (M-1) as a free sulfonic acid; 50 By using the procedure described in Step-2 of Example-1, Step-4: trans-sulfuric acid mono-2-(N' (R)-piperi and by using trans-3-N'-(6-benzyloxy-7-oxo-1,6-diaza-bi din-3-carbonyl-hydrazinocarbonyl)-7-oxo-1,6- cyclo[3.2.1]octane-2-carbonyl)-hydrazinocarbonyl-(R)- diaza-bicyclo[3.2.1]oct-6-ylester pyrrolidin-1-carboxylic acid tert-butyl ester (11.5gm, 0.024 55 mol), methanol (115 ml) and 10% palladium on carbon (3.0 By using the procedure described in Step-4 of Example-1 gm) to provide the title compound in 9.5gm quantity as a pale above, and by using tetrabutylammonium salt of trans-3-N'- brown solid and it was used for the next reaction immediately. (6-sulfooxy-7-oxo-1,6-diaza-bicyclo[3.2.1]octane-2-carbo Analysis: MS (ES+) CHNO398.2 (M+1): nyl)-hydrazinocarbonyl-(R)-piperidin-1-carboxylic acid tert-butyl ester (24 gm, 0.032 mmol), dichloromethane (60 60 Step-3: Preparation of tetrabutylammonium salt of ml) and trifluoroacetic acid (60 nil) to provide the title com trans-3-N'-(6-sulfooxy-7-oxo-1,6-diaza-bicyclo pound in 10gm quantity as a white solid, in 79% yield. 3.2.1]octane-2-carbonyl)-hydrazinocarbonyl-(R)- Analysis: MS (ES-)-CHNO,S-390.2 (M-1) as a pyrrolidin-1-carboxylic acid tert-butyl ester free sulfonic acid; 65 H'NMR (DMSO-d)=9.97 (d. 2H), 8.32(brs, 2H), 4.00 (br By using the procedure described in Step-3 of Example-1, s, 1H), 3.81 (d. 1H), 3.10-3.22 (m, 3H), 2.97-3.02 (m, 2H), and by using trans-3-N'-(6-hydroxy-7-oxo-1,6-diaza-bicy US 8,822,450 B2 29 30 clo3.2.1]octane-2-carbonyl)-hydrazinocarbonyl-(R)-pyr (4.0 gm, 9.73 mmol), obtained in Step-2 of Example-2) was rolidin-1-carboxylic acid tert-butyl ester (9.5gm, 0.024 mol), dissolved in DMF (12 ml) and to the clear solution was added pyridine (95 ml) and pyridine sulfur trioxide complex (19.08 potassium carbonate (1.61 gm, 11.6 mmol) followed by ethyl gm, 0.12 mol), 0.5 N aqueous potassium dihydrogen phos bromo acetate (1.2 ml, 10.0 mmol) under stirring and the phate solution (300 ml) and tetrabutylammonium sulphate suspension was stirred for 18 hours at about 25° C. The (8.15 gm, 0.024 mol) to provide the title compound in 15.3 reaction was monitored by TLC. DMF was evaporated under gm quantity as a yellowish Solid, in 87% yield. vacuum to provide a residue. The residue was purified by Analysis: MS (ES-) C.H.N.O.S.N(CH) as a silica gel column chromatography to provide titled Step-1 salt=476.1 (M-1) as a free sulfonic acid; 10 intermediate compound in 2.6 gm quantity as a solid in 53.7% H'NMR (DMSO-d)=9.82 (d. 2H), 3.97 (brs, 1H), 3.79 (d. yield. 1H), 3.42-3.44 (m, 1H), 3.00-3.18 (m. 10H), 2.65-2.97 (m, Analysis: MS (+)-CHNO=498.1 (M+1): 2H), 1.98-2.01 (m, 2H), 1.74-1.83 (m, 2H), 1.63-1.72 (m, 1H), 1.38-1.55 (m, 9H), 1.33 (s, 9H), 1.24-1.28 (m. 8H), H'NMR (CDC1)=8.45 (brs, 2H), 4.58 (s. 2H), 4.19-4.27 0.91-0.99 (m, 12H). 15 (m. 2H), 4.02-4.12 (m, 2H), 3.25 (brid, 1H), 3.15 (brd, 1H), 2.38 (brs, 1H), 2.35 (dd. 1H), 2.15-2.20 (m, 1H), 1.79-2.02 Step-4: trans-sulfuric acid mono-2-(N' (R)-pyrro (m, 4H), 1.67-1.77 (m, 4H), 1.44-1.51 (m. 11H), 1.28 (t,3H). lidin-3-carbonyl-hydrazinocarbonyl)-7-oxo-1,6- diaza-bicyclo[3.2.1]oct-6-ylester Step-2: Preparation of trans-3-N'-(6-car boxymethoxy-7-oxo-1,6-diaza-bicyclo3.2.1]octane By using the procedure described in Step-4 of Example-1, 2-carbonyl)-hydrazinocarbonyl-(R)-piperidin-1- and by using tetrabutylammonium salt of trans-3-N'-(6-sul carboxylic acid tert-butyl ester fooxy-7-oxo-1,6-diaza-bicyclo[3.2.1]octane-2-carbonyl)- hydrazinocarbonyl-(R)-pyrrolidin-1-carboxylic acid tert 25 To a clear solution of trans-3-N'-(6-ethoxycarbonyl butyl ester (15 gm, 0.021 mmol), dichloromethane (37 ml) methoxy-7-oxo-1,6-diaza-bicyclo[3.2.1]octane-2-carbo and trifluoroacetic acid (37 nil) to provide the title compound nyl)-hydrazinocarbonyl-(R)-piperidin-1-carboxylic acid in 7.7gm quantity as a white Solid. tert-butyl ester (600 mg, 1.20 mmol) in tetrahydrofuran (32

Analysis: MS (ES-)-CHNO,S-376.1 (M-1) as a 30 ml) and water (12 ml) was added lithium hydroxide (43.2 mg, free sulfonic acid; 1.8 mmol) at 0°C. The reaction mixture was stirred for 3 H'NMR (DMSO-d)=10.04 (s, 1H), 9.96 (s, 1H), 8.79 (br hours and was neutralized to pH 6 by addition of aqueous 1 N s, 1H), 8.68 (brs, 1H), 4.00 (brs, 1H), 3.82 (d. 1H), 3.18-3.32 potassium hydrogen Sulfate. It was exacted with ethyl acetate (m, 4H), 3.08-3.12 (m. 1H), 3.00 (brd, 1H), 2.05-2.29 (m, (3x25 ml). Layers were separated and aqueous layer was 1H), 1.96-2.05 (m, 2H), 1.84-1.87 (m. 1H), 1.69-1.73 (m, 35 acidified with 1 N potassium hydrogen sulfate to pH1 and 1H), 1.56-1.67 (m, 1H). extracted with ethyl acetate (3x25ml). The Organic layer was CI-44.2°, (c. 0.5, water). dried over Sodium sulfate and evaporated to dryness under Example-4 vacuum to provide 160 mg of tilted intermediate as a solid in 40 27% yield. trans-7-oxo-2-N ((R)-piperidin-3-carbonyl)- Analysis: MS (ES+)=CHNO-470.1 (M+1) hydrazinocarbonyl-1,6-diaza-bicyclo[3.2.1-oct-6- H'NMR (CDC1)=8.40 (brs, 2H), 4.67 (d. 1H), 4.52 (d. yloxy)-acetic acid 1H), 4.07-4.14 (m, 2H), 3.95 (brs, 1H), 3.43 (brd, 1H), 3.19 (brid, 1H), 2.47 (brs, 1H), 2.39 (dd. 1H), 2.09-2.13 (m, 2H), 45 1.77-2.00 (m, 4H), 1.68-1.77 (m, 2H), 1.45-1.51 (m. 11H). Step-3: trans-7-oxo-2-N'-((R)-piperidin-3-carbo nyl)-hydrazinocarbonyl-1,6-diaza-bicyclo[3.2.1- 50 oct-6-yloxy)-acetic acid To a clear solution of trans-3-N'-(6-carboxymethoxy-7- OXO-1,6-diaza-bicyclo3.2.1]octane-2-carbonyl)-hydrazi

55 nocarbonyl-(R)-piperidin-1-carboxylic acid tert-butyl ester (150 mg, 0.32 mmol) in dichloromethane (2 ml) was added trifluoroacetic acid under stirring at -10° C. The reaction mixture was stirred for about 1 hour at -10° C. and the Step-1: Preparation of trans-3-N'-(6-ethoxycarbon Solvents were evaporated under vacuum to provide a residue. ylmethoxy-7-oxo-1,6-diaza-bicyclo[3.2.1]octane-2- 60 The residue was triturated successively with diethyl ether (25 carbonyl)-hydrazinocarbonyl-(R)-piperidin-1-car ml) and acetonitrile (25 ml) and solvents were decanted to boxylic acid tert-butyl ester provide solid that was dried under vacuum to provide 59 mg of titled compound in 50% yield. The intermediate compound trans-3-N'-(6-hydroxy-7- 65 Analysis: MS (ES-)-CHNO368.0 (M-1) oXo-1,6-diaza-bicyclo3.2.1]octane-2-carbonyl)-hydrazi H'NMR (DMSO-d)=9.97 (brs, 2H), 4.48 (d. 1H), 4.29 (d. nocarbonyl-(R)-piperidin-1-carboxylic acid tert-butyl ester 1H), 3.91 (s, 1H), 3.83 (d. 1H), 3.36 (q, 1H), 3.11-3.21 (m,

US 8,822,450 B2 41 42 Example-43 aqueous layer was then washed with dichloromethane (25 mlx2) and layers were separated. The aqueous layer was Sodium salt of trans-sulfuric acid mono-2-(N'-(cy concentrated under vacuum below 40°C. to provide residue, ano-acetyl)-hydrazinocarbonyl)-7-oxo-1,6-diaza which was azeotroped with acetone and triturated with bicyclo[3.2.1]oct-6-ylester diethyl ether to provide a Suspension. The Suspension was filtered to provide title compound (Sodium salt of trans-sul furic acid mono-2-(N'-(2-cyano-acetyl)-hydrazinocarbo nyl)-7-oxo-1,6-diaza-bicyclo3.2.1]oct-6-ylester) in 300 mg O quantity and in 81% yield. H ul 10 Analysis: MS (ES-) CHNO.S.Na, 346.2 (M-1) as a --> H 'o. freeH'NMR sulfonic (DMSO-d)=10.2 acid; (s, 1H), 10.05 (s, 1H), 3.99 (s. O N 1H), 3.82 (d. 1H), 3.72 (s, 1H), 3.36 (s, 1H), 3.14 (brid, 1H), 2.99 (d. 1H), 1.98-2.03 (m. 1H), 1.75-1.84 (m, 1H), 1.56-1.72 O Noso Na 15 (m. 2H). 3 Compounds 44 to 52 (Table 2) were using a procedure described in Example-16 and using corresponding Tetrabutylammonium salt of trans-sulfuric acid mono-2- RCONHNH, in the place of cyano acetic acid hydrazide. (N'-(cyano-acetyl)-hydrazinocarbonyl)-7-oxo-1,6-diaza-bi cyclo[3.2.1]oct-6-ylester (600 mg. obtained using procedure 20 described in Step-1 to Step-3 of Example 1 (using cyano O acetic acid hydrazide in the place of (S)- N-tert-butoxycar- l bonyl-pyrrolidine-2-carboxylic acid hydrazide) was loaded R11 NN ... in tetrahydrofuran and water 1:9 mixture (10 ml) and passed H slowly through freshly activated Amberlite 200C resin in 25 N sodium form (100 gm). The column was eluted with 10% u1 tetrahydrofuran in water mixture. The fractions were ana- O YOSO,Na lyzed on TLC and desired fractions were evaporated to remove volatile solvent under vacuum below 40° C. The TABLE 2 Example Acid hydrazide Mass (ES-1) as No. (RCONHNH) R H NMR (DMSO-d) free acid (MF) 44. t-Boc NHNH t-C4H9 O—CO— 9.70 (s, 1H), 8.71 (s, 1H), 3.98 379.3 (s, 1H), 3.73, (d. 1H), 3.13 (d. (C2HNOSNa) 1H), 2.97 (brid, 1H), 1.97 2.01 (m, 1H), 1.73-1.83 (m, 1H), 155-2.72 (m, 2H), 1.38 (s, 9H).

45. 10.02 (brs, 2H), 3.98 (brs, 4O6.3 r O r O 1H), 3.85 (d. 1H), 3.66-2.70 (CHNOSNa) (m, 4H), 3.37 (brs, 2H), 3.05 Null NHNH2 - a (s,2.04 2H), (m, 2.76 1H), (brs, 1.73-1.82 4H), 2.00 (m, H), 1.58-1.70 (m, 2H).

46. 11.00 3H), 7.74 (s, 1H), 4.04 (brs, 1C2 NHNH2 -Ox2 8.911H), 3.91E.Y.S. (d. 1H), 3.06 E.(br chnos) HNCO N H2NCO N d, 1H), 1.90-2.06 (m, 2H), 1.77-1.87 (im, 1H), 1.61-1.77 O O (m, 2H).

47. O O 10.45 (brs, 1H), 10.08 (brs, 419.9 1H), 3.99 (brs, 1H), 3.81 (brid, (CHNOSNa) NHNH 1H), 3.56-3.58 (m, 4H), 341 r 1X 2H),3.51 2.06(m, (brs,2H), 2.99-3.102H) 2.01-2.03 (m, su O su O (m, 1H), 2.82-1.83 (m, 1H), 1.59-1.72 (m, 2H). 48. 9.81 (brs, 1H), 9.69 (brs, 1H), 419.2 5.77 (s, 2H), 4.21 (t, 1H), (CHNOSNa) NHNH 3.98 (brs, 1H), 3.77 (d. 1H), N 3.34-3.38 (m, 1H), 3.16-3.29 (m, 2H), 2.96 (brid, 1H), 1.73 NX 1.98 (m, 6H), 1.58-1.72 1s. 1. O (m, 2H). US 8,822,450 B2

TABLE 2-continued Example Acid hydrazide Mass (ES-1) as No. (RCONHNH) R H NMR (DMSO-d) free acid (MF) 49. F F 9.77 (brs, 2H), 5.95 (s. 2H), 437.2 5.32 (s, 0.5H), 5.18 (brs (CHNOFSNa) 0.5H), 4.38 (dd, 1H), 3.98 (br s, 1H), 3.81 (d. 1H), 3.35 NHNH2 3.59 (m, 2H), 2.97-3.25 (m, N N 1H), 2.96 (bird, 1H), 1.99 2.39 (m, 2H), 1.82-1.95 (m, O O 1H), 1.75-1.82 (m, 1H), O NH2 O NH2 1.59-1.69 (m, 2H). SO. 9.83 (brs, 2H), 7.32 (s, 1H), 454.2 4.21 (dd, 1H), 3.98 (brs, 1H), (CHNO.S.Na) NHNH 3.77 (d. 1H), 3.38-3.44 (m, N 2 N 1H), 3.21-3.25 (m, 1H), 2.90 s 2.98 ("s-l- 1's O o72.S. so O \ ... . S1. H3C CH3 H3C CH3 DO exchange: 3.95-4.05 (m, 374.2 2H), 3.10-3.19 (m, 1H), (C2HNO2S Na) NHNH2 2.95-3.05 (m, 1H), 1.68-1.98 NC NC (m, 4H), 1.29 (s.3H), 1.21 (s, 3H). O O 52. 9.95 (s, 2H), 7.93 (s, 1H), 4.03 O O 4.05 (m, 1H), 3.99 (brs, 1H), NHNH 3.81 (d. 1H), 3.19 (d. 1H), 2.98 N 2 N (brid, 1H), 2.28–2.48 (m, 1H), 1.84-2.25 (m, 5H), 1.56-1.75 boc O O (m, 2H).

Compounds of the invention from Example 1 to 52 were strains (NCTC 13351, M 50 and 7 MP) expressing ESBL prepared using (S)-pyrroglutamic acid as a starting com (Extended Spectrum Beta Lactamases). pound. The absolute stereochemistry is therefore (2S,5R) 35 7-OXO-1,6-diaza-bicyclo3.2.1]octane ring. Thus, the com TABLE 3 pound of Example-2, trans-sulfuric acid mono-2-(N'-((R)- piperidin-3-carbonyl-hydrazinocarbonyl)-7-OXO-1,6-diaza Antibacterial activity of representative compounds according bicyclo[3.2.1]oct-6-ylester has the absolute stereochemistry to the invention (MIC expressed in meg/ml as (2S,5R)-sulfuric acid mono-2-(N' (R)-piperidin-3-car 40 Bacterial Strain bonyl-hydrazinocarbonyl)-7-oxo-1,6-diaza-bicyclo3.2.1 Compound of E. coi E. coi E. coi oct-6-ylester. Alternatively, if the starting compound used is Sr. Example No. NCTC 13351 MSO 7 MP (R)-pyrroglutamic acid the resulting compounds will have (2R,5S) stereochemistry in 7-oxo-1,6-diaza-bicyclo[3.2.1 1. 1 O.O2S O.25 O.25 45 2. 2 O.25 O.25 1 octane ring. A reference to a compound according to the 3. 3 O.25 O.25 1 invention also includes corresponding compounds having 4. 4 8 8 16 5. 5 O.25 O.25 O.S (2S.5R) and (2R,5S) stereochemistry. 6. 6 32 64 128 7. 7 2 2 4 Biological Activity 8. 8 2 50 9. 9 2 4 10. 10 8 32 The biological activity of representative compounds 11. 11 2 4 according to the invention against various bacterial strains 12. 12 8 32 was investigated. In a typical study, overnightgrown bacterial 13. 13 2 14. 14 O.S O.25 2 cultures were diluted appropriately and inoculated on the agar 55 1S. 15 8 4 16 media containing doubling dilutions of the test compounds. 16. 16 2 Observation for growth or no growth was performed after 17. 17 O.S O.S 2 18. 18 8 4 32 16-20 hours of incubation at 35+2° C. in ambient air. The 19. 19 2 overall procedure was performed as per Clinical and Labora 20. 2O 2 2 8 tory Standards Institute (CLSI) recommendations (Clinical 60 21. 21 4 and Laboratory Standards Institute (CLSI), Performance 22. 22 4 4 8 Standards for Antimicrobial Susceptibility Testing, 20th 23. 23 4 4 8 24. 24 4 Informational Supplement, M 100-S20, Volume 30, No. 1, 25. 25 4 2010). The results of these studies are summarized in Tables 26. 26 2 3 to 8. 65 27. 27 4 2 8 Table 3 details antibacterial activity of representative com 28. 28 4 pounds according to the invention against various E. coli US 8,822,450 B2 45 46 TABLE 3-continued The antibacterial activity of representative compounds according to the invention was also investigated in combina Antibacterial activity of representative compounds according tion with at least one antibacterial agent using the above study to the invention (MIC expressed in meg/ml protocol and the results are given Table 6. As can be seen, the Bacterial Strain use of compounds according to the invention significantly Compound of E. coi E. coi E. coi lowered MIC values of the antibacterial agent (e.g. in this case Sr. Example No. NCTC 13351 MSO 7 MP Ceftazidime). The results also suggest the compounds according the invention increase antibacterial effectiveness of 29. 29 32 16 128 the antibacterial agent when said antibacterial agent is co 30. 30 16 8 32 10 31. 31 1 1 4 administered with a pharmaceutically effective amount of a 32. 32 O.S O.S 1 compound of Formula (I) or a stereoisomer or a pharmaceu 33. 33 1 O.S 2 tically acceptable salt thereof. 34. 34 2 2 8 35. 35 1 1 4 The antibacterial activity of representative compounds 36. 36 1 1 2 15 according to the invention was also investigated in combina 37. 37 2 2 4 tion with a beta-lactamase inhibitor using the above study 38. 38 4 2 8 protocol and the results are given in Table 7. As can be seen, 39. 39 4 2 8 40. 40 1 1 4 the compounds according to the invention in combination 41. 41 4 4 16 with a beta lactam inhibitor exhibited excellent antibacterial 42. 42 32 32 64 activity against various bacterial strains. For example, a com 43. 43 1 2 8 bination comprising compound of Example 2 (and also 44. 44 32 32 64 45. 45 4 8 8 Example 3) according to the invention in combination with 46. 46 32 64 128 sulbactam exhibited much superior MIC values compared 47. 47 32 64 128 with when these were used alone. The results also suggest that 48. 48 16 8 32 compounds according to the invention in combination with a 49. 49 32 16 16 25 SO. 50 16 16 32 beta-lactam inhibitor can be effectively used in preventing or S1. 51 16 16 32 treating a bacterial infection in a subject, including those 52. 52 O.S O.S 2 infections caused by bacteria producing one or more beta lactamase enzymes. The antibacterial activity of representative compounds Tables 4 and 5 provide antibacterial activity of representa 30 tive compounds according to the invention against various according to the invention was also investigated in combina Multi Drug Resistant (MDR) Gram-negative bacterial strains tion with a beta-lactamase inhibitor and a antibacterial agent expressing various ESBLS. The activities are expressed as using the above study protocol and the results are given in MICs (mcg/ml). For comparison, the activity of several Table 8. AS can be seen, the compounds according to the known antibacterial agents (for example, Ceftazidime, AZtre invention in combination with at least one beta lactam inhibi onam, Imipenem, Ciprofloxacin and Tigecycline) are also 35 tor and at least one antibacterial agent exhibited excellent included. As can be seen, the representative compounds antibacterial activity against various bacterial Strains. For according to the invention exhibit antibacterial activity example, a combination comprising compound of Example 2 against various MDR strains. The data in Table 4 and 5 also (and also Example 3) according to the invention in combina indicates that the compounds according to the invention tion with sulbactam and Cefepime exhibited better MIC val exhibit potent activity against a wide variety of bacteria, even 40 ues compared with when these were used alone. The results against those producing different types of beta-lactamase also suggest that compounds according to the invention in enzymes. In general, the activity of the compounds according combination with at least one beta-lactam inhibitor and at to the invention against various beta-lactamase producing least one antibacterial agent can be effectively used in pre bacterial strains is even better than the other antibacterial venting or treating a bacterial infection in a Subject, including agents currently employed in the clinical practice to treat Such 45 those infections caused by bacteria producing one or more infections. beta-lactamase enzymes. TABLE 4 Comparative antibacterial activity of representative compounds according to the invention against various Multi Drug Resistant (MDR) Gran negative strains (expressed as MICs (meg/ml). Class AESBL Class C ESBL KPCESBL

E. Coi E. Coi E. Coi E. Coi E. Coi E. Coli Kpenumoniae Kpenumoniae Kpenumoniae Sr. Compound W 13353 W 13351 W 13352 MSO H 484 B 89 HS21 HS23 H525

1. Ceftazidime 32 32 >32 >32 >32 >32 >32 >32 >32 2. Aztreonam >32 >32 >32 >32 >32 >32 >32 >32 >32 3. Imipenem O.25 O.25 O.25 O.S 4 O.S 16 16 16 4. Ciprofloxacin >32 O.S O.12 >32 >32 >32 32 8 32 5. Tigecyclin 1 1 O.25 O.S O.25 O.S 2 8 2 6. Example 1 O.25 O.S O.S O.S 2 O.S O.S O.S O.S 7. Example 2 O.12 O.25 O.25 O.25 O.12 O.25 O.S 1 O.S 8. Example 3 O.25 O.S O.S O.S O.S O.25 O.S O.S O.S 9. Example 5 O.S O.25 O.S 1 1 O.S 1 1 2 10. Example 7 O.S 2 1 2 4 4 2 1 2 11. Example 8 1 1 1 1 2 1 1 O.S 1 12. Example 9 1 1 1 1 2 1 1 O.S O.S 13. Example 11 2 2 2 2 16 2 4 2 2 US 8,822,450 B2 47 48 TABLE 4-continued Comparative antibacterial activity of representative compounds according to the invention against various Multi Drug Resistant (MDR) Gran negative strains (expressed as MICs (meg/ml). Class AESBL Class C ESBL KPCESBL E. Coi E. Coi E. Coi E. Coi E. Coi E. Coi Kpenumoniae Kpenumoniae Kpenumoniae Sr. Compound W 13353 W 13351 W 13352 MSO H484 B 89 HS21 HS23 H525 14. Example 13 1 1 2 1 4 1 1 O.S 1 15. Example 14 O.S O.S O.S O.S 2 O.S O.S O.25 O.25 16. Example 17 O.25 O.S O.S O.S 4 O.S O.S O.S O.S 17. Example 19 2 2 2 2 4 1 1 1 1 18. Example 30 8 16 16 8 8 8 8 8 >32 19. Example 35 1 1 1 1 1 1 2 1 1 20. Example 38 2 4 4 2 8 2 2 2 2 21. Example 43 1 1 1 1 8 1 2 1 1

TABLE 5 Comparative antibacterial activity of representative compounds according to the invention against various Multi Drug Resistant (MDR) Gran negative strains (expressed as MICs (meg/ml). Class BESBL K. penumoniae E. Coi E. Coli F aeruginosa Sr. Compound S 48 M3 M44 ATCC 27853 PS21 PS32

1. Ceftazidime >32 >32 >32 1 >32 >32 2. Aztreonam >32 >32 >32 2 8 8 3. Imipenem 16 8 32 4 >32 >32 4. Ciprofloxacin >32 >32 >32 O.S 32 O.12 5. Tigecyclin 1 4 O.25 16 16 16 6. Example 1 2 1 0.25 16 16 32 7. Example 2 O.S 2 O.S 8 8 8 8. Example 3 O.S O.S O.12 2 4 4 9. Example 5 2 1 O.S >32 >32 >32 10. Example 7 4 4 2 >32 >32 >32 11. Example 8 1 4 O.S 8 16 8 12. Example 9 1 2 O.S 8 8 16 13. Example 11 2 8 1 >32 >32 >32 14. Example 13 1 1 1 32 32 16 15. Example 14 4 O.S O.25 8 16 8 16. Example 17 O.S O.S O.S >32 >32 >32 17. Example 19 4 1 1 32 >32 >32 18. Example 30 >32 8 16 >32 >32 >32 19. Example 35 2 1 2 >32 >32 32 2O. Example 38 4 1 4 >32 >32 >32 21. Example 43 1 4 O.S >32 >32 >32

TABLE 6 TABLE 6-continued Antibacterial activity of Ceftazidime in presence of representative compounds of the invention against various 50 Antibacterial activity of Ceftazidime in presence of Multi Drug Resistant (MDR) Gran negative strains. representative compounds of the invention against various Multi Drug Resistant (MDR) Gram negative strains. Ceftazidine MIC (expressed in mcg/ml Ceftazidime MIC (expressed in mcg/ml) K. pneumoniae P. vulgaris ATCC 700603 S-137B (ESBL type: (ESBL type: 55 K. pneumoniae P. vulgaris Sr. Composition Class A) Class C) ATCC 700603 S-137B (ESBL type: (ESBL type: 1. Ceftazidime alone >32 >32 Sr. Composition Class A) Class C) 2. Ceftazidime + Compound O.O6 1 of Example 1 (4 mcg/ml) 3. Ceftazidime + Compound O.O6 1 60 7. Ceftazideftazidime + UompounC d O.12 1 of Example 2 (4 mcg/ml) of Example 9 (4 mcg/ml) 4. Ceftazidime + Compound O.O6 1 8. Ceftazidime + Compound O.O3 O.S of Example 3 (4 mcg/ml) of Example 14 (4 mcg.mil) 5. Ceftazidime + Compound O.25 4 of Example 5 (4 mcg/ml) Note: 6. Ceftazidime + Compound O.O6 O.S 65 The MICs of each of compounds of Example 1,2,3,5,8,9 and 14 when used alone (in the of Example 8 (4 mcg/ml) absence of Ceftazidime) is >32 mcg.mil. US 8,822,450 B2 49 50 TABLE 7 Antibacterial activity of Sulbactan in combination with a compound according to the invention. MIC of sulbactan (expressed in mcg/ml A. battmannii A. battmannii A. battmannii A. battmannii Sr. Composition J-143 146.0648 S-334 G-16S 1. Sulbactam alone 32 32 32 32 2. Sullbactam + compound of Example 2 (4 mcg/ml) 4 4 4 2 3. Sullbactam + compound of Example 2 (8 mcg/ml) 4 4 2 2 4. Sulbactam + compound of Example 3 (4 mcg/ml) 8 4 8 2 5. Sullbactam + compound of Example 3 (8 mcg/ml) 1 2 4 2 Standalone MIC of compound of Example 2 and Example 3 for each of the strains was >32 mcg.mil.

TABLE 8 Antibacterial activity of an antibacterial agent in combination with Sulbactan and a compound according to the invention. MIC of Cefepine (expressed in mcg/ml

A. battmannii A. battmannii A. battmannii A. battmannii S . Composition J-143 1460648 S-334 G-16S

Cefepime alone >32 >32 32 >32 Cefepime + compound of Example 2 (4 mcg/ml) >32 32 32 >32 Cefepime + compound of Example 2 (8 mcg/ml) 32 32 32 32 Cefepime + Sulbactam (4 mcg/ml) 16 32 16 16 Cefepime + Sulbactam (8 mcg/ml) 16 16 8 8 Cefepime + Sulbactam (8 mcg/ml) + compound of Example 2 (4 mcg/ml) O.25 O.25 O.25 O.25 Cefepime + Sulbactam (8 mcg/ml) + compound of Example 2 (8 mcg/ml) O.25 O.25 O.25 O.25 Cefepime + compound of Example 3 (4 mcg/ml) >32 32 32 >32 Cefepime + compound of Example 3 (8 mcg/ml) 32 32 32 32 Cefepime + Sulbactam (8 mcg/ml) + compound of Example 3 (4 mcg/ml) O.12 O.12 O.12 O.12 Cefepime + Sulbactam (8 mcg/ml) + compound of Example 3 (8 mcg/ml) O.O6 O.O6 O.O6 O.O6 Standalone MIC of sulbactam for each of the strains was 32 mcgfml. Standalone MIC of compound of Example 2 and Example 3 for each of the strains was >32 mcg.mil.

35 The invention claimed is: M is hydrogen or a cation; 1. A compound of Formula (I) R is: (a) hydrogen, (b) C-C alkyl optionally substituted with one or more Formula (I) 40 Substituents independently selected from halogen, OR, CN, COOR, CONRR, NRR, NRCORs, NRCONRR-7, heterocyclyl, heteroaryl, cycloalkyl or aryl, (c) CN, N R2 (d) NRR-7, Ou- No1 45 (e) CONRR, (f) NHCONRR, or a stereoisomer or a pharmaceutically acceptable salt (g) aryl optionally substituted with one or more Substitu thereof 50 ents independently selected from C-C alkyl, ORs, NRR, halogen, CN, CONRR-7, SO-alkyl, SO wherein: aryl, OSO-alkyl, OSO-aryl, or NHCONRR, R is: (h) heterocyclyl optionally substituted with one or more (a) hydrogen, Substituents independently selected from C-C alkyl, (b) (CO), R., or 55 ORs, NRR, halogen, CN, CONRR, SO-alkyl, (c) COOR. SO-aryl, OSO-alkyl, OSO-aryl, or NHCONRR, n is 0, 1 or 2; (i) heteroaryl optionally substituted with one or more R is: Substituents independently selected from C-C alkyl, (a) SOM, 60 ORs, NRR-7, halogen, CN, CONRR-7, SO-alkyl, (b) SONH, SO-aryl, OSO-alkyl, OSO-aryl, or NHCONRR, (j) cycloalkyl optionally substituted with one or more (c) POM, Substituents independently selected from C-C alkyl, (d) CHCOOM, ORs, NRR-7, halogen, CN, CONRR-7, SO-alkyl, (e) CFCOOM, 65 SO-aryl, OSO-alkyl, OSO-aryl, or NHCONRR, (f) CHFCOOM, or (k) cycloalkyl substituted with C-C alkyl wherein (g) CF; C-C alkyl is further substituted with one or more US 8,822,450 B2 51 52 substituents independently selected from ORs, trans-Sulfuric acid mono-2-(N'-(amino-acetyl)-hydrazi NRR, halogen, CN, or CONRR, or nocarbonyl)-7-oxo-1,6-diaza-bicyclo[3.2.1]oct-6-yl) (i) ORs: ester, R is: trans-Sulfuric acid mono-2-(N'-(3-amino-propioyl)-hy (a) hydrogen, drazinocarbonyl)-7-oxo-1,6-diaza-bicyclo[3.2.1]oct-6- (b) C-C alkyl optionally substituted with one or more ylester; Substituents independently selected from halogen, trans-Sulfuric acid mono-2-(N'-(4-amino-butanoyl)-hy OR, CN, COOR, CONRR, NRR, NRCORs, drazinocarbonyl)-7-oxo-1,6-diaza-bicyclo[3.2.1]oct-6- heterocyclyl, heteroaryl, cycloalkyl or aryl, ylester; (c) aryl optionally substituted with one or more substitu 10 trans-sulfuric acid mono-2-(N'-((2S)-2-amino-3-hy ents independently selected from C-C alkyl, ORs, droxy-propioyl)-hydrazinocarbonyl)-7-oxo-1,6-diaza NRR, halogen, CN, CONRR-7, SO-alkyl, SO bicyclo[3.2.1]oct-6-ylester; aryl, OSO-alkyl, OSO-aryl, or NHCONRR, trans-sulfuric acid mono-2-(N'-(2S,4S)-4-fluoro-pyrroli (d) heterocyclyl optionally substituted with one or more 15 din-2-carbonyl-hydrazinocarbonyl)-7-oxo-1,6-diaza Substituents independently selected from C-C alkyl, bicyclo[3.2.1]oct-6-ylester; ORs, NRR-7, halogen, CN, CONRR-7, SO-alkyl, trans-sulfuric acid mono-2-(N'-(2S,4R)-4-methoxy-pyr SO-aryl, OSO-alkyl, OSO-aryl, or NHCONRR, rolidin-2-carbonyl-hydrazinocarbonyl)-7-oxo-1,6- (e) heteroaryl optionally substituted with one or more diaza-bicyclo[3.2.1]oct-6-ylester; Substituents independently selected from C-C alkyl, trans-Sulfuric acid mono-2-(N'-(piperidin-4-carbonyl)- ORs, NRR, halogen, CN, CONRR, SO-alkyl, hydrazinocarbonyl)-7-oxo-1,6-diaza-bicyclo[3.2.1]oct SO-aryl, OSO-alkyl, OSO-aryl, or NHCONRR, 6-ylester; O trans-sulfuric acid mono-2-(N' -((RS)-piperidin-3-car (f) cycloalkyl optionally substituted with one or more bonyl)-hydrazinocarbonyl)-7-oxo-1,6-diaza-bicyclo Substituents independently selected from C-C alkyl, 25 3.2.1]oct-6-ylester; ORs, NRR-7, halogen, CN, CONRR-7, SO-alkyl, trans-sulfuric acid mono-2-(N-((S)-piperidin-3-carbo SO-aryl, OSO-alkyl, OSO-aryl, or NHCONRR: nyl)-hydrazinocarbonyl)-7-oxo-1,6-diaza-bicyclo Rs and Rs are each independently: 3.2.1]oct-6-ylester; (a) hydrogen, or trans-sulfuric acid mono-2-(N' -((RS)-piperidin-2-car (b) C-C alkyl optionally substituted with one or more 30 bonyl)-hydrazinocarbonyl)-7-oxo-1,6-diaza-bicyclo Substituents independently selected from halogen, 3.2.1]oct-6-ylester; CN, CONRR, NRR, heterocyclyl, heteroaryl, trans-sulfuric acid mono-2-(N-(S)-piperidin-2-carbo cycloalkyl or aryl; nyl)-hydrazinocarbonyl)-7-oxo-1,6-diaza-bicyclo Re and R7 are each independently: 3.2.1]oct-6-ylester; (a) hydrogen, 35 trans-sulfuric acid mono-2-(N' ((R)-piperidin-2-carbo (b) C-C alkyl optionally substituted with one or more nyl)-hydrazinocarbonyl)-7-oxo-1,6-diaza-bicyclo Substituents independently selected from halogen, 3.2.1]oct-6-ylester; ORs, CN, COORs, CONRRs, NRRs, NRCORs, trans-Sulfuric acid mono-2-(N'-(piperazin-4-yl-acetyl)- heterocyclyl, heteroaryl, cycloalkyl or aryl, hydrazinocarbonyl)-7-oxo-1,6-diaza-bicyclo[3.2.1]oct (c) aryl optionally substituted with one or more substitu 40 6-ylester; ents independently selected from C-C alkyl, ORs, trans-sulfuric acid mono-2-(N' ((RS)-1-amino-1-phe NRRs halogen, CN, CONRRs, SO-alkyl, SO nyl-acetyl)-hydrazinocarbonyl)-7-oxo-1,6-diaza-bicy aryl, OSO-alkyl, OSO-aryl, or NHCONRRs. clo3.2.1]oct-6-ylester; (d) heterocyclyl optionally substituted with one or more trans-sulfuric acid mono-2-(N'-(3-amino-2,2-dimethyl Substituents independently selected from C-C alkyl, 45 propioyl)-hydrazinocarbonyl)-7-OXO-1,6-diaza-bicyclo ORs, NRSRs, halogen, CN, CONRRs, SO-alkyl, 3.2.1]oct-6-ylester; SO-aryl, OSO-alkyl, OSO-aryl, or NHCONRRs, trans-Sulfuric acid mono-2-(N'-(1-aminomethyl-cyclo (e) heteroaryl optionally substituted with one or more propan-1-carbonyl)-hydrazinocarbonyl)-7-oxo-1,6- Substituents independently selected from C-C alkyl, diaza-bicyclo[3.2.1]oct-6-ylester; ORs, NRSRs, halogen, CN, CONRRs, SO-alkyl, 50 trans-Sulfuric acid mono-2-(N'-(5-amino-pentanoyl)-hy SO-aryl, OSO-alkyl, OSO-aryl, or NHCONRRs, drazinocarbonyl)-7-oxo-1,6-diaza-bicyclo[3.2.1]oct-6- (f) cycloalkyl optionally substituted with one or more ylester; Substituents independently selected from C-C alkyl, trans-sulfuric acid mono-2-(N' (R)-pyrrolidin-2-car ORs, NRRs halogen, CN, CONRRs, SO-alkyl, bonyl-hydrazinocarbonyl)-7-oxo-1,6-diaza-bicyclo SO-aryl, OSO-alkyl, OSO-aryl, or NHCONRRs, 55 3.2.1]oct-6-ylester; O trans-Sulfuric acid mono-2-(N'-(2-amino-thiazol-4-car (g) Re and R, are joined together to form a four to seven bonyl)-hydrazinocarbonyl)-7-oxo-1,6-diaza-bicyclo member ring. 3.2.1]oct-6-ylester; 2. A compound according claim 1, selected from: or a stereoisomer or a pharmaceutically acceptable salt trans-sulfuric acid mono-2-(N' (S)-pyrrolidin-2-carbo 60 thereof. nyl-hydrazinocarbonyl)-7-oxo-1,6-diaza-bicyclo 3. A compound according claim 1, selected from: 3.2.1]oct-6-ylester; Sodium salt of trans-sulfuric acid mono-2-(N'-(cyano trans-sulfuric acid mono-2-(N-((R)-piperidin-3-carbo acetyl)-hydrazinocarbonyl)-7-oxo-1,6-diaza-bicyclo nyl)-hydrazinocarbonyl)-7-oxo-1,6-diaza-bicyclo 3.2.1]oct-6-ylester; 3.2.1]oct-6-ylester; 65 Sodium salt of trans-N'-(7-oxo-6-sulfooxy-1,6-diaza-bi trans-Sulfuric acid mono-2-hydrazinocarbonyl-7-oxo-1, cyclo3.2.1]octane-2-carboxylic acid)-hydrazinecar 6-diaza-bicyclo3.2.1]oct-6-ylester; boxylic acid tert-butyl ester; US 8,822,450 B2 53 54 Sodium salt of trans-sulfuric acid mono-2-(N'-(morpho 9. A pharmaceutical composition according to claim 6. lin-4-yl-acetyl)-hydrazinocarbonyl)-7-oxo-1,6-diaza wherein said antibacterial agent is selected from a group bicyclo[3.2.1]oct-6-ylester; consisting of penicillins, penems, carbapenems, cephalospor Sodium salt of trans-sulfuric acid mono-2-(N'-(6-car boxamido-pyridin-2-carbonyl)-hydrazinocarbonyl)-7- ins, and monobactams. oxo-1,6-diaza-bicyclo[3.2.1]oct-6-ylester; 10. A pharmaceutical composition according to claim 6. Sodium salt of trans-sulfuric acid mono-2-(N'-(morpho wherein the antibacterial agent is a cephalosporin antibiotic lin-4-oxo-carbonyl)-hydrazinocarbonyl)-7-OXO-1,6- selected from a group consisting of cephalothin, cephalori diaza-bicyclo[3.2.1]oct-6-ylester; dine, cefaclor, cefadroxil, cefamandole, cefazolin, cephal Sodium salt of trans-sulfuric acid mono-2-(N' (S)-1- exin, cephradine, ceftizoxime, cefoxitin, cephacetrile, cefo carbamoyl-pyrrolidin-2-carbonyl-hydrazinocarbonyl)- 10 tiam, cefotaxime, cefsulodin, cefoperaZone, ceftizoxime, 7-oxo-1,6-diaza-bicyclo[3.2.1]oct-6-ylester; cefnenoxime, cefimetazole, cephaloglycin, cefonicid, cefo Sodium salt of trans-sulfuric acid mono-2-(N'-(2S4S)- dizime, cefpirome, ceftazidime, ceifriaxone, cefpiramide, 1-carbamoyl-4-fluoro-pyrrolidin-2-carbonyl-hydrazi cefbuperaZone, cefoZopran, cefepime, cefoselis, ceflupre nocarbonyl)-7-oxo-1,6-diaza-bicyclo[3.2.1]oct-6-yl) nam, cefuZonam, cefpimizole, cefclidin, cefixime, ceftibuten, ester, 15 cefdinir, cefpodoxime axetil, cefpodoxime proxetil, cefteram Sodium salt of trans-sulfuric acid mono-2-(N' (S)-1- methanesulfonyl-pyrrolidin-2-carbonyl-hydrazinocar pivoxil, cefetamet pivoxil, cefcapene pivoxil or cefditoren bonyl)-7-oxo-1,6-diaza-bicyclo[3.2.1]oct-6-ylester; pivoxil, cefuroxime, cefuroxime axetil, loracarbacef, ceftaro Sodium salt of trans-sulfuric acid mono-2-(N'-(cyano line and latamoxef. dimethyl-acetyl)-hydrazinocarbonyl)-7-oxo-1,6-diaza 11. A pharmaceutical composition according to claim 6. bicyclo[3.2.1]oct-6-ylester; wherein the antibacterial agent is selected from a group con Sodium salt of trans-sulfuric acid mono-2-(N' (S)-5- sisting of ceftazidime, cefepime, cefpirome, piperacillin oXo-pyrrolidin-2-carbonyl-hydrazinocarbonyl)-7-OXo doripenem, meropenem, imipenem, ceftaroline and ceftolo 1,6-diaza-bicyclo[3.2.1]oct-6-ylester; ZaC. or a stereoisomer thereof. 25 12. A pharmaceutical composition according to claim 6. 4. A pharmaceutical composition comprising a compound wherein the antibacterial agent is selected from a group con according to any of the claims 1 to 3. sisting of aminoglycosides, ansamycins, carbacephems, 5. A pharmaceutical composition according to claim 4. cephalosporins, cephamycins, lincosamides, lipopeptides, further comprising at least one beta-lactamase inhibitor macrollides, monobactams, nitrofurans, penicillins, polypep selected from Sulbactam, taZobactam, clavulanic acid, or a 30 tides, quinolones, Sulfonamides, tetracyclines, or oxazolidi pharmaceutically acceptable derivative thereof. none antibacterial agents. 6. A pharmaceutical composition according to claim 4 or 5. 13. A pharmaceutical composition according to claim 5, further comprising at least one antibacterial agent or a phar comprising (a) trans-Sulfuric acid mono-2-(N' (R)-piperi maceutically acceptable derivative thereof. din-3-carbonyl-hydrazinocarbonyl)-7-oxo-1,6-diaza-bicy 7. A pharmaceutical composition according to claim 6. 35 clo3.2.1]oct-6-yl ester or a stereoisomer or a pharmaceuti wherein the antibacterial agent is selected from a group con cally acceptable salt thereof, and (b) sulbactam or a sisting of aminoglycosides, ansamycins, carbacephems, pharmaceutically acceptable derivative thereof. cephalosporins, cephamycins, lincosamides, lipopeptides, 14. A pharmaceutical composition according to claim 5. macrollides, monobactams, nitrofurans, penicillins, polypep comprising (a) trans-Sulfuric acid mono-2-(N'-(R)-pyrro tides, quinolones, Sulfonamides, tetracyclines, or oxazolidi 40 lidin-3-carbonyl-hydrazinocarbonyl)-7-oxo-1,6-diaza-bi none antibacterial agents. cyclo3.2.1]oct-6-yl ester or a stereoisomer or a pharmaceu 8. A pharmaceutical composition according to claim 6. tically acceptable salt thereof, and (b) sulbactam or a wherein the antibacterial agent is a beta-lactam antibacterial pharmaceutically acceptable derivative thereof. agent. k k k k k