EA042632B1 - DIAZABICLOCTANONES AS SERINE BETA-LACTAMASE INHIBITORS - Google Patents

Description

Field of invention

The invention relates to compounds that are inhibitors of serine beta-lactamase enzymes, and compositions containing such compounds. These compounds are useful in the treatment of bacterial infection. The invention also relates to combinations of these compounds with other active agents useful in the treatment of bacterial infection.

Prior Art

The resistance of pathogenic bacteria to antibiotics is a serious threat to public health worldwide. Of particular concern is bacterial resistance to β-lactam antibiotics due to hydrolysis of β-lactam antibiotics by β-lactamase enzymes. These enzymes catalyze the hydrolytic cleavage of the β-lactam ring, rendering the β-lactam antibiotic inactive. Beta-lactamases belong to two families of enzymes that differ in structure and mechanism of action: serine β-lactamases (SBL (from the English serine β-lactamases); classes A, C and D), in which the serine residue in the active center is used for cleavage β-lactam by the mechanism of covalent catalysis, and metallo-β-lactamases (MBL (from the English metallo β-lactamases); class B), in the case of which the mechanism of catalysis by metal ions is used for direct hydrolysis of β-lactam without the formation of a covalent intermediate.

To counter the threat of resistance, the natural product clavulanic acid from Streptomyces clavuligerus (an SBL inhibitor: see Scheme 1 in the General Synthesis Methodology section below) was introduced in 1981 as part of a co-combination with the β-lactam antibiotic amoxicillin (called Augmentin) (See De Koning, G. A. et al., 1981, J. Antimicrobial Chemotherapy, 8, 81-82). Recently, there has been renewed interest in the discovery of β-lactamase inhibitors in order to counter the threat from new β-lactamases that are not inhibited by clavulanic acid, such as extended spectrum β-lactamases (ESBL) and carbapenemases. . This has led to the development of new classes of synthetic inhibitors such as diazabicyclooctonanes (DBOs) of which avibactam is clinically used in combination with ceftazidime (Mawal, Y. et al., 2015, Expert. Rev. Clin. Pharmacol., 8 (6), 691-707).

Carbapenems such as meropenem and imipenem are broadly regarded as the drugs of choice for the treatment of severe infections caused by ESBL-producing enterobacteria and Acinetobacter baumannii. Although avibactam is a good nanomolar inhibitor of many clinically relevant SBLs that catalyze the hydrolysis of carbapenems, it is not effective against (1) variants of the OXA family (from the English oxacillin - oxacillin), which are the most common carbapenemases in Europe and on Middle East; and especially with respect to (2) OXA-producing strains of Acinetobacter baumannii (declared by the World Health Organization as a major priority pathogen) (Canton R. et al., European Network on Carbapenemases, (2012), Rapid evolution and spread of carbapenemases among Enterobacteriaceae in Europe. , Clin. Microbiol. Infect., 18: 413-431 and Nordmann P. et al., 2011, Global spread of carbapenemase-producing Enterobacteriaceae, Emerg. Infect. Dis., 17: 1791-1798).

Thus, there is a need to develop new SBL inhibitors, in particular inhibitors capable of inhibiting the β-lactamases of the OXA family. There is also a need to develop a broad spectrum of inhibitors capable of inhibiting a range of SBLs, including extended spectrum β-lactamases (ESBLs) and carbapenemases. In addition, there is a need to develop inhibitors capable of inhibiting SBL produced by Acinetobacter baumannii. The present invention is directed to solving some or all of these problems.

Brief description of the essence of the invention

The present inventors surprisingly found that the new DBO analogs have unexpected activity against SBL enzymes, including OXA variants. In this regard, it is expected that these compounds will find use as additives to carbapenems in the treatment of infectious disease.

Accordingly, the present invention provides a compound as described in aspect [1]. These compounds are useful in the treatment of bacterial infection, in particular they are useful in eliminating or reducing SBL-mediated bacterial resistance to antibiotics. Thus, they are useful in the treatment of infection caused by SBL-producing bacteria, for example, in combination with antibiotics. Thus, the treatment or prevention of bacterial infections is generally carried out by administering a compound of the invention in combination with an antibiotic. The compounds are also useful in combination with metallo-β-lactamase (MBL) inhibitors, particularly when the bacterial infection is caused by both SBL- and MBL-producing bacteria.

The present invention provides the following.

1. A compound that is diazabicyclooctanone of formula (I) or a pharmaceutically acceptable salt thereof,

- 1 042632

FORMULA (I) where

R is selected from halogen, C _1-4 alkyl and LXR ¹ , wherein the C _1-4 alkyl group is substituted with at least one halogen atom and optionally additionally substituted with one or two R ² substituents;

R ¹ is C _1-4 alkyl which is substituted with at least one halogen atom and optionally additionally substituted with one or two R ² substituents;

each R ² is independently a thiazole;

L is a bond or is a C _1-2 alkylene group which is unsubstituted or substituted by at least one halogen atom; And

X is O or S(O)z, where z is 0.

2. A compound as defined above, wherein said compound is diazabicyclooctanone of formula (II) or a pharmaceutically acceptable salt thereof, or diazabicyclooctanone of formula (III) or a pharmaceutically acceptable salt thereof

Formula (II) Formula (III) where R is halogen.

3. A compound as defined above, wherein R is a fluorine or chlorine atom.

4. A compound as defined above, selected from (2R,5R)-2-fluoro-7-oxo-1,6-diazabicyclo[3.2.1]octan-6-yl-hydrosulfate;

(2S,5R)-2-fluoro-7-oxo-1,6-diazabicyclo[3.2.1]octan-6-yl-hydrosulfate;

(2R,5R)-2-chloro-7-oxo-1,6-diazabicyclo[3.2.1]octan-6-yl-hydrosulfate;

and their pharmaceutically acceptable salts.

5. A compound as defined above, which is diazabicyclooctanone of formula (II) or a pharmaceutically acceptable salt thereof,

FORMULA (II) where R is as defined above in claim 1.

6. A compound as defined above, which is diazabicyclooctanone of formula (II) or a pharmaceutically acceptable salt thereof,

FORMULA (II) where

R is C _1-4 alkyl, wherein the C _1-4 alkyl group is substituted with at least one

- 2 042632 a halogen atom and optionally substituted with one or two R ² substituents;

each R ² is independently a thiazole.

7. Connection as defined above, where

1) R is C _1-4 alkyl, wherein the C _1-4 alkyl group is substituted with at least one halogen atom and optionally additionally substituted with one R ² substituent; or

2) R ¹ is C _1-2 alkyl, wherein the C _1-2 alkyl group is substituted with at least two halogen atoms selected from fluorine and chlorine atoms; or

3) said compound is as defined in claim 5 and R is LXR ¹ where

L is a bond or is an unsubstituted C1 alkylene group; X is O or S; And

R ¹ is a C1 alkyl group substituted with 1, 2 or 3 halogen groups; or

4) R is selected from -CH ₂ -O-CF ₃ and -S-CF3; or

5) R is selected from CF ₃ , CHF ₂ , CHCl ₂ , CCI ₃ , CH ₂ F, CF ₂ CH ₃ , CF ₂ -thiazolyl and CH ₂ CF ₃ , preferably from CF 3 , CHF 2 and CHCl.

8. In particular, the compound of the invention is (2R,5R)-2-fluoro-7-oxo-1,6diazabicyclo[3.2.1]octan-6-yl hydrogen sulfate or a pharmaceutically acceptable salt thereof.

9. In particular, the compound of the invention is sodium (2R,5R)-2-fluoro-7-oxo-1,6diazabicyclo[3.2.1]octan-6-yl sulfate.

10. A pharmaceutical composition containing a compound of any invention, a pharmaceutically acceptable carrier or diluent.

11. The pharmaceutical composition may further comprise a β-lactam antibiotic.

12. The pharmaceutical composition as described above, wherein the β-lactam antibiotic is specifically meropenem.

13. Combination of a compound of the invention and a β-lactam antibiotic for the treatment or prevention of a bacterial infection.

14. Combination according to the invention, where the β-lactam antibiotic is meropenem.

15. The use of a compound, composition or combination as described above to eliminate or reduce antibiotic resistance in Gram-negative bacteria.

16. Use as described above, wherein the Gram-negative bacteria are selected from Enterobacteria, Pseudomonas and Moraxella.

17. Use as described above, wherein the bacteria selected from Enterobacteria, Pseudomonas and Moraxella are selected from Klebsiella pneumoniae, Escherichia coli, Enterobacter cloacae, Pseudomonas aeruginosa, Burkholderia cepacia and Acinetobacter baumannii.

18. The use of a compound, composition or combination described above for the treatment or prevention of a bacterial infection.

19. Use as described above, comprising the use of a compound of the invention for the treatment or prevention of a bacterial infection by co-administration with a β-lactam antibiotic.

20. Use as described above, wherein the β-lactam antibiotic is meropenem.

21. Use as described above, wherein the bacterial infection is caused by bacteria selected from Enterobacteria, Pseudomonas and Moraxella.

22. Use as described above, wherein the bacteria selected from Enterobacteria, Pseudomonas and Moraxella are selected from Klebsiella pneumoniae, Escherichia coli, Enterobacter cloacae, Pseudomonas aeruginosa, Burkholderia cepacia and Acinetobacter baumannii.

23. Use as described above, wherein the bacterial infection is caused by carbapenem-resistant enterobacteria.

Brief description of the figures

In FIG. 1 shows MIC (minimal inhibiting concentration) data (described in the examples section) demonstrating the activity of various inhibitors against two panels (Fig. 1A and B) of clinical strains of OXA-positive enterobacteria resistant to beta- lactam antibiotics such as meropenem, cefepime and ceftazidime. Data are shown for (1) meropenem monotherapy (MEM, from meropenem);

(2) clinical combination of the VNRX-5133 inhibitor and the antibiotic cefepime (CEF(cefepime)/VNRX);

(3) clinical combination of the inhibitor avibactam and the antibiotic ceftazidime (CAZ/AVI, from ceftazidime/avibactam); and (4) the compound of example 10 and the antibiotic meropenem (MEM + compound of example 10).

In FIG. Figure 2 shows MIC data (described in the examples section) demonstrating the activity of various inhibitors against a panel of carbapenemase-positive Klebsiella pneumoniae (KPC) clinical strains of enterobacteria resistant to

- 3 042632 to beta-lactam antibiotics such as meropenem, cefepime and ceftazidime. Data are shown for (1) meropenem as monotherapy (MEM);

(2) clinical combination of the VNRX-5133 inhibitor and the antibiotic cefepime (CEF/VNRX);

(3) clinical combination of the inhibitor avibactam and the antibiotic ceftazidime (CAZ/AVI); and (4) the compound of example 10 and the antibiotic meropenem (MEM + compound of example 10).

In FIG. 3 shows MIC data (described in the examples section) demonstrating the activity of various inhibitors against Acinetobacter baumannii. Data are shown for (1) meropenem as monotherapy (MEM);

(2) clinical combination of the VNRX-5133 inhibitor and the antibiotic cefepime (CEF/VNRX);

(3) clinical combination of the inhibitor avibactam and the antibiotic ceftazidime (CAZ/AVI); and (4) the compound of example 10 and the antibiotic meropenem (MEM + compound of example 10).

In FIG. 4 shows scatterograms showing the bacterial load (colony forming units (CFU)/g) in mouse thigh tissue after infection:

(A) KPC positive strain of K. pneumoniae NR-48977;

(B) OXA-positive K. pneumoniae AC00783; or (C) OXA-positive A. baumannii AC00445, in each case after 9 hours; and treatment with meropenem as monotherapy or in combination with the compound of example 10, as indicated on the x-axis.

The geometric mean for bacterial load for each treatment is indicated by the horizontal bar, and the reduction in bacterial load in thigh tissue (in log ₁₀ ) compared to meropenem alone is indicated in brackets. Statistical significance is determined in comparison with meropenem as monotherapy (**p<0.01;***p<0.001).

Detailed description of the invention

Definitions

As used herein, a C1- _C6 alkyl group is a linear or branched alkyl group containing from 1 to 6 carbon atoms. A C1- _C4 alkyl group is a linear or branched alkyl group containing 1 to 4 carbon atoms. A C1- _C4 alkyl group is often a C1- _C3 alkyl group or a _C2 - _C4 alkyl group. Examples of C1- _C4 alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl and tert-butyl. The C ₁ -C ₂ alkyl group is methyl or ethyl, usually methyl. For the avoidance of doubt, if two alkyl groups are present, these alkyl groups may be the same or different.

As used herein, a C1- _C2 alkylene group is an unsubstituted or substituted bidentate group resulting from the removal of two hydrogen atoms from a C1- _C2 alkane. These two hydrogen atoms can be removed from the same carbon atom or from different carbon atoms. The C1- _C2 alkylene group is methylene or ethylene, usually methylene.

As used in this description, C ₁ . ₄ alkoxy is a _{C1_4} alkyl group as defined above attached to an oxygen atom.

As used herein, a C ₂ -C ₄ alkenyl group is a linear or branched alkenyl group containing 2 to 4 carbon atoms and having one or more, eg one or two, usually one, double bond. Typically, the C ₂ -C ₄ alkenyl group is a C ₂ -C ₃ alkenyl group. Examples of C ₂ -C ₄ alkenyl groups include ethenyl, propenyl and butenyl. A C2-C4 alkenylene group is an unsubstituted or substituted bidentate group resulting from the removal of two hydrogen atoms from a C2-C4 alkene. Typically, the C ₂ -C ₄ alkenylene group is a C ₂ -C ₃ alkenylene group, for example ethenylene, propenylene or butenylene.

As used herein, a C2-C4 alkynyl group is a linear or branched alkynyl group containing 2 to 4 carbon atoms and having one or more, eg one or two, usually one triple bond. Typically, the C ₂ -C ₄ alkynyl group is a C ₂ -C ₃ alkynyl group, for example ethynyl, propynyl or butynyl.

The C ₂ -C ₄ alkynylene group is an unsubstituted or substituted bidentate group resulting from the removal of two hydrogen atoms from a C ₂ -C ₄ alkyne. Typically the C ₂ -C ₄ alkynylene group is a C ₂ -C ₃ alkynylene group, for example ethynylene, propynylene or butynylene.

An alkyl, alkylene, alkoxy, alkenyl, alkynyl, or alkynylene group as used herein may be unsubstituted or substituted. Unless otherwise indicated, substituted alkyl, alkylene, alkoxy, alkenyl, alkenylene, alkynyl or alkynylene groups generally contain one or more, such as 1, 2, 3 or 4, such as one, two or three, such as two or three deputies. Preferred substituents are halogen atoms. Where indicated, the alkyl group may also be substituted with one or two R ² groups, as defined herein. The substituents themselves on a substituted alkyl or alkoxy group

- 4 042632 are generally not substituted unless otherwise indicated. If more than one substituent is present, they may be the same or different.

As used herein, halogen is usually chlorine, fluorine, bromine or iodine, and is preferably chlorine, bromine or fluorine, especially chlorine or fluorine, in the most particular case fluorine. When a group or moiety is substituted with one or more halogen atoms, it preferably carries one, two, three or four halogen atoms, preferably two or three halogen atoms. When a group or moiety is substituted with two or more halogen atoms, the halogen atoms may be the same or different. Usually, the halogen atoms are the same.

As used herein, a 6-10 membered aryl group is a substituted or unsubstituted monocyclic or fused polycyclic aromatic group containing 6 to 10 carbon atoms in the ring portion. Examples include monocyclic groups such as phenyl and fused bicyclic groups such as naphthyl and indenyl. A preferred group is phenyl (benzene).

As used herein, a 5-10 membered heteroaryl group is a substituted or unsubstituted monocyclic aromatic group containing 5 to 10 atoms in the ring portion, including at least one heteroatom, e.g. 1, 2, or 3 heteroatoms, usually selected from O, S and N; and is usually a 5-6 membered heteroaryl group which is a substituted or unsubstituted monocyclic aromatic group containing 5 or 6 atoms in the ring moiety, including at least one heteroatom, e.g. 1, 2 or 3 heteroatoms, usually selected from O , S and N. Examples of 5- and 6-membered heteroaryl groups include pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyridine, pyridazine, pyrimidine and pyrazine.

As used herein, a 3-10 membered heterocyclic group is a cyclic group containing 3 to 10 atoms selected from C, O, N and S in the ring, including at least one heteroatom and usually one or two heteroatoms ; and is typically a 3-8 membered heterocyclic group, typically a 4-6 membered heterocyclic group, which is a cyclic group containing 4 to 6 atoms selected from C, O, N, and S in the ring, including at least one heteroatom and usually one or two heteroatoms. Typically, the heteroatom or heteroatoms are(s) selected from O, N and S. The heterocyclic group may be saturated or partially unsaturated. A 4-6 membered partially unsaturated heterocyclic group is a cyclic group containing 4 to 6 atoms selected from C, O, N and S in the ring and containing 1 or 2, for example 1 double bond.

A 3-10 membered carbocyclyl group is a cyclic hydrocarbon containing 3 to 10 carbon atoms. The carbocyclyl group may be saturated or partially unsaturated, but is usually saturated. A 3-10 membered partially unsaturated carbocyclyl group is a cyclic hydrocarbon containing 3 to 10 carbon atoms and containing 1 or 2, for example 1 double bond. A 3-10 membered carbocyclyl group is usually a 410 membered carbocyclyl group, for example a 3-8 membered carbocyclyl group such as a 3-6-, 4-6-membered or 5-6-membered carbocyclic group. Examples of 3-6 membered saturated carbocyclic groups include cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl groups.

Examples of 4-6 membered saturated heterocyclic groups include oxetane, azetidine, piperazine, piperidine, morpholine, pyrrolidine, imidazolidine and oxazolidine. Examples of 4-6 membered partially unsaturated heterocyclic groups include tetrahydropyrazine, tetrahydropyridine, dihydro-1,4oxazine, tetrahydropyrimidine, dihydro-1,3-oxazine, dihydropyrrole, dihydroimidazole and dihydrooxazole.

An aryl, heterocyclyl or heteroaryl group may be unsubstituted or substituted as described in this application. For example, an aryl, heterocyclyl or heteroaryl group may be unsubstituted or substituted with 1, 2 or 3, usually 1 or 2 substituents, such as 1 substituent. Suitable substituents include halogen, OH, C(O) _{C1_4} alkyl, C(O)OH, C(O) _OC1-4 alkyl, _C1 . ₄ αlkyl and C ₁ . ₄ alkoxy, while themselves C ₁ . ₄ αalkyl and C ₁ . The ₄ alkoxy groups and moieties are unsubstituted or substituted by one or more halogen atoms.

As used herein, a pharmaceutically acceptable salt is a salt with a pharmaceutically acceptable acid or base. Pharmaceutically acceptable acids include both inorganic acids such as hydrochloric, sulfuric, phosphoric, pyrophosphoric, hydrobromic or nitric acid and organic acids such as oxalic, citric, fumaric, maleic, malic, ascorbic, succinic, tartaric, benzoic, acetic, methanesulfonic, ethanesulfonic, benzenesulfonic or p-toluenesulfonic acid. Pharmaceutically acceptable bases include alkali metal (eg sodium or potassium) and alkaline earth metal (eg calcium or magnesium) hydroxides and organic bases such as alkylamines, aralkylamines and heterocyclic amines. Preferred pharmaceutically acceptable salts are those derived from the SO3H group with pharmaceutically acceptable bases, in particular quaternary ammonium salts, eg tetrabutylammonium salts, or alkali metal salts, eg sodium or potassium salts, most preferably sodium salts.

- 5 042632

In formula (I), the bicyclic ring assumes the depicted stereochemical configuration.

Thus, if the bicyclic ring is shown in the form of a chair (as shown below), then the R group is in the axial up position or in the equatorial down position, while the second ring, bearing the OSO3H substituent, is in the axial down position.

In formula (II), the bicyclic ring assumes the depicted stereochemical configuration. Thus, if the bicyclic ring is depicted as a chair (as shown below), then the R group is in the axial up position, while the second ring bearing the OSO ₃ H substituent is in the axial down position.

In formula (III), the bicyclic ring assumes the depicted stereochemical configuration. Thus, if the bicyclic ring is shown in the form of a chair (as shown below), then the R group is in the equatorial down position, while the second ring, bearing the OSO3H substituent, is in the axial down position.

Typically, a compound or composition described herein contains at least 50%, preferably at least 60%, 75%, 90%, or 95% of a compound of formula (I) having a stereochemical configuration at the bicyclic ring (i.e., relatively two chiral centers in a bicyclic ring) shown above. Preferably, the compound is substantially diastereomerically pure with respect to the two chiral centers depicted above.

Usually, if the compound or composition described (described) in this application contains a compound of formula (II), then at least 50%, preferably at least 60, 75, 90 or 95% of said compound has a stereochemical configuration at the bicyclic ring ( i.e. relative to the two chiral centers in the bicyclic ring) shown above. Preferably, the compound is substantially diastereomerically pure with respect to the two chiral centers depicted above.

Typically, if a compound or composition described (described) in this application contains a compound of formula (III), then at least 50%, preferably at least 60%, 75%, 90% or 95% of said compound has a stereochemical configuration at the bicyclic ring (i.e. relative to the two chiral centers in the bicyclic ring) shown above. Preferably, the compound is substantially diastereomerically pure with respect to the two chiral centers depicted above.

In formula (I), (II) or (III), one or more additional chiral centers may be present in the R group. Regarding these chiral centers, the stereochemical configuration is given without limitation, and such compounds can be used in diastereomeric pure form or in form of a mixture of isomers. Usually, the compound or composition described(s) in this application contains at least 50%, preferably at least 60%, 75%, 90% or 95% of the compound corresponding to formula (I), (II) or (III), which is diastereomeric pure with respect to a chiral center in the R group. Typically, a compound or composition of the invention contains at least 60% by weight, such as at least 75%, 90%, or 95% of the individual diastereomer. Preferably, the compound is substantially optically pure.

Typically, a compound of formula (I) is a compound of formula (II).

In addition, for the avoidance of ambiguity, the compounds of the invention may be used in any tautomeric form.

As used herein, the elimination or reduction of antibiotic resistance, or the elimination or reduction of resistance in bacteria, means that there is a prevention or suppression of the manifestation of the mechanism of bacterial resistance, for example, cleavage of the antibiotic by SBL. Thus, there is an elimination or reduction of the effects of bacterial resistance (i.e., the ineffectiveness of antibiotics). In other words, the compounds of the invention are useful for inhibiting or preventing hydrolysis of the β-lactam ring, i.e. to inhibit or prevent hydrolysis of the antibiotic. Thus, they improve the efficacy of antibiotics when used to treat an infection caused by SBL-producing bacteria.

Compounds of the invention

In some preferred compounds of formula (I), R is halogen. Preferably, if R is halogen, then R is a fluorine or chlorine atom. Most preferably, if R is a halogen then R is a fluorine atom.

When R is halogen, the compound of formula (I) may be a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a compound of formula (III) or a pharmaceutically acceptable salt thereof,

Formula (II) Formula (III) where R is halogen.

Preferably, if R is halogen, then the compound of formula (I) may be a compound of formula (II) or a pharmaceutically acceptable salt thereof.

Thus, preferably, the compound is a compound of formula (II) or a pharmaceutically acceptable salt thereof, or a compound of formula (III) or a pharmaceutically acceptable salt thereof, and R represents a fluorine or chlorine atom. More preferably, the compound is a compound of formula (II) or a pharmaceutically acceptable salt thereof, and R is a fluorine or chlorine atom. Most preferably, the compound is a compound of formula (II) or a pharmaceutically acceptable salt thereof, and R is a fluorine atom.

Preferably, in the present invention, the compound of formula (I) is a compound of formula (II) or a pharmaceutically acceptable salt thereof.

In the case of other preferred compounds, the compound of formula (I) is a compound of formula (II) or a pharmaceutically acceptable salt thereof; Where

R is selected from C(O)R1, C _1-4 alkyl and LXR ¹ , wherein the C _1-4 alkyl group is substituted with at least one halogen atom and optionally additionally substituted with one or two R ² substituents;

R ¹ is C _1-4 alkyl which is substituted with at least one halogen atom and optionally additionally substituted with one or two R ² substituents;

each R ² is independently selected from OH; C _1-4 alkoxy which is unsubstituted or substituted by one or more halogen atoms; C(O)R ³ ; C(O)OH; C(O)OR ³ ; 6-10 membered aryl; 5-6 membered heteroaryl; and 4-6 membered heterocyclyl;

while aryl, heteroaryl and heterocyclyl groups are unsubstituted or substituted by one, two or three substituents selected from halogen, OH, C(O)R ³ , C(O)OH, C(O)OR ³ and C1 ₄ alkyl and C _1-4 alkoxy groups which are themselves unsubstituted or substituted by one or more halogen atoms;

R ³ is C _1-4 alkyl which is unsubstituted or substituted by one or more halogen atoms;

L is a bond or is a C _1-2 alkylene group which is unsubstituted or substituted by at least one halogen atom; And

X is O or S(O)Z, where z is 0, 1 or 2.

If R is LXR ¹ , then L is preferably a bond or an unsubstituted C _1-2 alkylene group. More preferably, L is a bond or is

- 7 042632 is an unsubstituted C1 alkylene (methylene) group. X is selected from O and S(O) _z , where z is 0, 1, or

2; those. X is selected from O, S, S(O) and S(O) ₂ . Preferably X is selected from O and S.

Preferred LXR ^r groups include -CH2-OR ¹ , -SR ¹ and -SO2-R ¹ where R ¹ is as defined herein. More preferred LXR ¹ groups include -CH2-OR ¹ and -SR ¹ where R ¹ is as defined herein. For example, in such groups, R ¹ is preferably a C1 or C2 alkyl group, preferably a C1 alkyl group. The group R1 is preferably substituted with 1, 2 or 3 halo groups, for example three halo groups; for example, the R ¹ group is preferably CF3. Thus, the most preferred LCR ¹ groups include -LX-CF3 groups such as -CH2-O-CF3 and -S-CF ₃ .

In the case of other preferred compounds, the compound of formula (I) is a compound of formula (II) or a pharmaceutically acceptable salt thereof; Where

R is selected from C(O)R ¹ and C _1-4 alkyl, wherein the C _1-4 alkyl group is substituted with at least one halogen atom and optionally additionally substituted with one or two R ² substituents;

R ¹ is C _1-4 alkyl which is substituted with at least one halogen atom and optionally additionally substituted with one or two R ² substituents;

each R ² is independently selected from OH; C1-4 alkoxy which is unsubstituted or substituted by one or more halogen atoms; C(O)R ³ ; C(O)OH; C(O)OR ³ ; 6-10 membered aryl; 5-6 membered heteroaryl; and 4-6 membered heterocyclyl; while aryl, heteroaryl and heterocyclyl groups are unsubstituted or substituted by one, two or three substituents selected from halogen, OH, C(O)R ³ , C(O)OH, C(O)OR ³ and C1-4 alkyl and C _1-4 alkoxy groups which are themselves unsubstituted or substituted by one or more halogen atoms;

R ³ is C _1-4 alkyl which is unsubstituted or substituted by one or more halogen atoms.

Preferably R is C ₁ . ₄ alkyl, while C ₁ . The _4αalkylα group R is as defined herein.

Typically, in such compounds, R is selected from C(O)R ¹ and C1.4 alkyl, with the C1.4 α alkyl α group substituted with at least one halogen atom (eg, fluorine or chlorine) and optionally additionally substituted with one R ² substituent. Preferably, R is selected from C(O)R ¹ and C1. ₂ alkyl, while C ₁ . _{The 2} αalkyl group is substituted with at least one halogen atom (eg fluorine or chlorine atom) and optionally additionally substituted with one R ² substituent.

According to one preferred aspect of the invention, the compound is a compound of formula (II) or a pharmaceutically acceptable salt thereof, and R is C _1-4 alkyl, wherein the C _1-4 alkyl group is substituted with at least one halogen atom (e.g., a fluorine atom or chlorine) and optionally additionally substituted with one or two substituents, for example one R ² substituent. Preferably, the alkyl group is substituted with at least two, for example two or three, halogen atoms. The halogen atoms are preferably selected from fluorine and chlorine atoms. Preferably, R is C1.2αkyl, wherein the C1.2alkyl group is substituted with at least one halogen atom (eg, fluorine or chlorine) and optionally additionally substituted with one R ² substituent. More preferably R is C1. ₂ alkyl, while C ₁ . _{The 2} alkylα group is substituted with at least two halogen atoms selected from fluorine and chlorine atoms. Most preferably R is a methyl group which is substituted with two or three halogen atoms selected from fluorine and chlorine atoms, eg CF ₃ , CCl ₃ , CHF 2 and CHCl 2 .

When R is C(O)R ¹ , then typically R ¹ is C1-4 alkyl which is substituted with at least one halogen atom (eg fluorine or chlorine atom) and optionally additionally substituted with one substituent selected from OH and C1. 4alkoxy which is itself unsubstituted or substituted by one or more halogen atoms (eg fluorine or chlorine atoms). Preferably, R ¹ is C1. ₄ alkyl which is substituted with at least one, more preferably at least two (eg two or three) halogen atoms. The halogen atoms are preferably selected from fluorine and chlorine atoms. More preferably R ¹ is C1.2alkyl, wherein the C1_2alkyl group is substituted with at least two halogen atoms selected from fluorine and chlorine atoms. Most preferably R ¹ is a methyl group which is substituted by two or three halogen atoms selected from fluorine and chlorine atoms, for example is CF3.

When R ² is a 6-10 membered aryl, it is usually phenyl. When R ² is a 5-6 membered heteroaryl, it is usually a 5 membered heteroaryl selected from thiazolyl, pyrrolyl, furanyl, thiophenyl, imidazolyl and oxazolyl, preferably thiazolyl. When R ² is a 4-6 membered heterocyclyl, it is usually a 4- or 5-membered heterocyclyl group selected from oxetanyl, azetidinyl, tetrahydrofuranyl, pyrrolidinyl and tetrahydrothiophenyl. Preferably, it is a 4-membered heterocyclyl group selected from oxetanyl and azetidinyl, most preferably oxetanyl.

If R ² is an aryl, heteroaryl or heterocyclyl group, then it is unsubstituted or substituted with one, two or three (for example, one or two) substituents selected from halogen, OH, C(O)R ³ , C (O)OH, C(O)OR ³ and C _1-4 alkyl and C _1-2 alkoxy groups which are themselves unsubstituted or substituted by one or more halogen atoms. Preferred substituents are halogen; HE; and C _1-2 alkyl and C _1-2 alkoxy groups which are themselves unsubstituted or substituted with one or more halogen atoms. More preferred substituents are halogen, OH, Me and OMe. Most preferably, if R ² represents an aryl, heteroaryl or heterocyclyl group, then it is unsubstituted.

Typically, each R ² is independently selected from OH; C1-2 alkoxy which is itself unsubstituted or substituted by one or more halogen atoms; C(O)OR ³ where R ³ is unsubstituted C1-2αkyl; and unsubstituted 5-6 membered heteroaryl. Preferably, each R ² is independently selected from OH; OME; C(O)OMe; and unsubstituted thiazolyl.

Typically R ³ is unsubstituted C _1-2 alkyl, preferably methyl.

In the case of certain preferred compounds of the invention, the compound is a compound of formula (II) or a pharmaceutically acceptable salt thereof, and

R is C _1-4 alkyl, wherein the C _1-4 alkyl group is substituted with at least one halogen atom and optionally additionally substituted with one or two R ² substituents; wherein R is preferably C _1-2 alkyl, wherein the C _1-2 alkyl group is substituted with at least one halogen atom (eg fluorine or chlorine atom) and optionally additionally substituted with one R ² substituent;

each R ² is independently selected from OH; C1-4alkoxy which is unsubstituted or substituted by one or more halogen atoms; C(O)R ³ ; C(O)OH; C(O)OR ³ ; 6-10 membered aryl; 5-6 membered heteroaryl; and 4-6 membered heterocyclyl; while the aryl, heteroaryl and heterocyclyl groups are unsubstituted or substituted by one, two or three substituents selected from halogen, OH, C(O)R ³ , C(O)OH, C(O)OR ³ and C1-4 alkyl and C _1-4 alkoxy groups which are themselves unsubstituted or substituted by one or more halogen atoms; wherein preferably each R ² is selected from OH; C _1-2 alkoxy which is itself unsubstituted or substituted by one or more halogen atoms; C(O)OR ³ and unsubstituted 5-6 membered heteroaryl; And

R ³ is C1-4 alkyl which is unsubstituted or substituted by one or more halogen atoms; preferably R ³ is unsubstituted C1-2 alkyl.

Other preferred compounds of the invention are those wherein the compound is a compound of formula (II) or a pharmaceutically acceptable salt thereof, and wherein

R is selected from C(O)R1 and C _1-4 alkyl, wherein the C _1-4 alkyl group is substituted with at least one halogen atom (eg fluorine or chlorine) and optionally additionally substituted with one R ² substituent;

R ¹ is C _1-4 alkyl which is substituted with at least one halogen atom (eg fluorine or chlorine atom) and optionally additionally substituted with one substituent selected from OH and C _1-4 alkoxy which is itself unsubstituted or substituted with one or several halogen atoms (for example, a fluorine or chlorine atom); And

R ² is selected from OH; C1-2alkoxy which is itself unsubstituted or substituted by one or more halogen atoms; C(O)OR ³ where R ³ is unsubstituted C1-2alkyl; and unsubstituted 5-6 membered heteroaryl.

More preferred compounds of the invention are those wherein the compound is a compound of formula (II) or a pharmaceutically acceptable salt thereof, and wherein

R is selected from C(O)R1 and C _1-4 alkyl, in particular C _1-2 alkyl, wherein the alkyl group is substituted with at least one halogen atom (eg fluorine or chlorine atom) and optionally additionally substituted with one substituent R ² ;

R ¹ is C _1-2 alkyl, wherein the C _1-2 alkyl group is substituted with at least two halogen atoms selected from fluorine and chlorine atoms; And

R ² is selected from OH; OME; C(O)OMe; and unsubstituted thiazolyl.

Even more preferred compounds of the invention are those wherein the compound is a compound of formula (II) or a pharmaceutically acceptable salt thereof, and wherein R is COCF3 or C _1-2 alkyl, wherein the C _1-2 alkyl group is substituted with at least one atom halogen (eg fluorine or chlorine atom) and optionally additionally substituted with one R ² substituent, R ² being selected from OH; OME; C(O)OMe; and unsubstituted thiazolyl. More preferably R is C _1-2 alkyl, wherein the C _1-2 alkyl group is substituted with at least two halogen atoms selected from fluorine or chlorine. Most preferably R is a methyl group which is substituted with two or three halogen atoms selected from fluorine and chlorine atoms, eg CF ₃ , CCI ₃ , CHF2 and CHC12.

Preferred examples of R groups include CF ₃ , CHF2, CHCl ₂ , CCl ₃ , CH2F, CF2CH3, CF2CH2CO2Me, COCF3, CF2-thiazolyl, CF2CH2OCH3, CF2CH2CH2OH, CH(OH)CF3, CH2CF3, CF2oxetanyl, especially CF3, CHF2 and CHCl2 .

Preferred compounds of the invention include

- 9 042632 (25^)-7-oxo-2-(trifluoromethyl)-1,6-diazabicyclo[3.2.1]octan-6-ylhydrosulfate;

(25^)-7-oxo-2-(difluoromethyl)-1,6-diazabicyclo[3.2.1]octan-6-ylhydrosulfate;

(25^)-2-(dichloromethyl)-7-oxo-1,6-diazabicyclo[3.2.1]octan-6-ylhydrosulfate;

(25^)-7-oxo-2-(trichloromethyl)-1,6-diazabicyclo[3.2.1]octan-6-ylhydrosulfate;

(25^)-2-(fluoromethyl)-7-oxo-1,6-diazabicyclo[3.2.1]octan-6-ylhydrosulfate;

(23^)-7-oxo-2-(trichloromethyl)-1,6-diazabicyclo[3.2.1]octan-6-ylhydrosulfate;

(2S,5R)-2-(1,1-diφtoroethyl)-7-oκco-1,6-diazabiL^iclo[3.2.1]octaan-6-ylhydrosulfate;

methyl 3,3-difluoro-3-((25.5H)-7-oxo-6-(sulfooxy)-1,6diazabicyclo[3.2.1]octan-2-yl)propanoate;

(25^)-7-oxo-2-(2,2,2-trifluoroacetyl)-1,6-diazabicyclo[3.2.1]octan-6-ylhydrosulfate;

(2S,5R)-2-[diφτop(1,3-thiazol-2-yl)methyl]-7-oxo-1,6d andazabicyclo[3.2.1]octan-6-yl-hydrosulfate;

(2S,5R)-2-(1,1-difluoro-2-methoxyethyl)-7-oxo-1,6-diazabicyclo[3.2.1]octan-6yl hydrogen sulfate;

(2S,5R)-2-(1,1-difluoro-2-hydroxyethyl)-7-oxo-1,6-diazabicyclo[3.2.1]octan6-ylhydrosulfate;

(25^)-7-oxo-2-(2,2,2-trifluoro-1-hydroxyethyl)-1,6d and azabicyclo[3.2.1]octan-6-yl-hydrosulfate;

(25^)-7-oxo-2-(2,2,2-trifluoroethyl)-1,6-diazabicyclo[3.2.1]octan-6-ylhydrosulfate;

(25^)-2-[difluoro(oxetan-3-yl)methyl]-7-oxo-1,6-diazabicyclo[3.2.1]octan6-yl hydrogen sulfate;

(25^)-2-(chloromethyl)-7-oxo-1,6-diazabicyclo[3.2.1]octan-6-ylhydrosulfate;

(2R,5R)-7-oκco-2-[(τpiφtormethyl)sulfanyl]-1,6-diazabicyclo[3.2.1]octaan6-yl-hydrosulfate;

(25.5P)-7-oxo-2-[(trifluoromethoxy)methyl]-1,6-diazabicyclo[3.2.1]octan-6yl hydrogen sulfate;

(2R,5R)-2-φτop-7-oκco-1,6-diazabicyclo[3.2.1]octaan-6-yl-hydrocylo;

(2S,5R)-2-φτop-7-oκco-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate; and (2R,5R)-2-chloro-7-oκco-1,6-diazabicyclo[3.2.1]octaan-6-yl-hydrocylφat;

and their pharmaceutically acceptable salts.

Preferred pharmaceutically acceptable salts of these compounds are those containing the SO3H group, in particular quaternary ammonium salts, eg tetrabutylammonium salts, and alkali metal salts, eg sodium or potassium salts. Most preferred are the sodium salts, which are listed below:

- 10 042632 sodium (2S,5R)-7-oκco-2-(τpiφτopmethyl)-1,6-diazabicyclo[3.2.1]octan-6 ylsulfate; sodium (25.5C)-7-oxo-2-(difluoromethyl)-1,6-diazabicyclo[3.2.1]octan-6yl sulfate; sodium (28^)-2-(dichloromethyl)-7-oxo-1,6-diazabicyclo[3.2.1]octan-6yl sulfate; sodium (28.5P)-7-oxo-2-(trichloromethyl)-1,6-diazabicyclo[3.2.1]octan-6 ylsulfate; sodium (28^)-2-(fluoromethyl)-7-oxo-1,6-diazabicyclo[3.2.1]octan-6yl sulfate; sodium (28^)-7-oxo-2-(trichloromethyl)-1,6-diazabicyclo[3.2.1]octan-6yl sulfate; sodium (2S,5R)-2-(1,1-d iffluoroethyl)-7-oxo-1,6-d andazabicyclo[3.2.1]octan-6ylsulfate;

methyl-(2S,5R)-3,3-diφτop-3-(7-oκco-6-{[(codiooκci)cylφonyl]oκci}-1,6diazabicyclo[3.2.1]octan-2-yl)propanoate;

sodium (2S,5R)-7-oκco-2-(2,2,2-τpiφtoroacetyl)-1,6-diazabicyclo[3.2.1]octaan 6-ylsulfate;

sodium (2S,5R)-2-[diφτop(1,3-thiazol-2-yl)methyl]-7-oxo-1,6d andazabicyclo[3.2.1]octan-6-ylsulfate; sodium (2S,5R)-2-(1,1-diφτop-2-methoκciethyl)-7-oκco-1,6diazabicyclo[3.2.1]octan-6-ylsulfate; sodium (2S,5R)-2-(1,1-diφτop-2-hydroxiethyl)-7-occo-1,6 d-iazabicyclo[3.2.1]octan-6-ylsulfate;

sodium (2S,5R)-7-oκco-2-(2,2,2-τpiφτop-1-hydropoκceethyl)-1,6d and azabicyclo[3.2.1]octan-6-ylsulfate; sodium (2S,5R)-7-oκco-2-(2,2,2-τpiφtoroethyl)-1,6-diazabicyclo[3.2.1]octaan-6 ylsulfate; sodium (2S,5R)-2-[diφτop(occeta- n-3-yl)methyl]-7-occo-1,6d andazabicyclo[3.2.1]octan-6-ylsulfate; sodium (28.5^-2-(chloromethyl)-7-oxo-1,6-diazabicyclo[3.2.1]octan-6 ylsulfate; sodium (2R,5R)-7-oκco-2-[(τpiφtormethyl) sulfanyl]-1,6d iazabicyclo[3.2.1 ]octan-6-ylsulfate; 6-ylsulfate, sodium (2R,5R)-2-φτop-7-oκco-1,6-diazabicyclo[3.2.1]octaan-6-ylcylφat, sodium (2S,5R)-2-φτop-7-oκco -1,6-diazabicyclo[3.2.1]octan-6-yl sulfate, and sodium (2R,5R)-2-chloro-7-oκco-1,6-diazabicyclo[3.2.1]octan-6-ylcylφaτ.

More preferred compounds are

- 11 042632 (2S,5R)-7-oκco-2-(τpiφτopmethyl)-1,6-diazabiL^iclo[3.2.1]octaan-6-ylhydrosulfate;

(2S,5R)-7-oκco-2-(diφtormethyl)-1,6-diazabicyclo[3.2.1]octaan-6-ylhydrosulfate;

(25.5K)-2-(dichloromethyl)-7-oxo-1,6-diazabicyclo[3.2.1]octan-6-ylhydrosulfate;

(25,5^-2-(fluoromethyl)-7-oxo-1,6-diazabicyclo[3.2.1]octan-6-ylhydrosulfate;

(28,5^-2-(1,1-difluoroethyl)-7-oxo-1,6-diazabicyclo[3.2.1]octan-6-ylhydrosulfate;

(2S,5R)-2-[diφτop(1,3-thiazol-2-yl)methyl]-7-oxo-1,6d andazabicyclo[3.2.1]octan-6-ylhydrosulphate;

(28,5^-2-(chloromethyl)-7-oxo-1,6-diazabicyclo[3.2.1]octan-6-ylhydrosulfate;

(2R,5R)-7-oκco-2-[(τpiφtormethyl)sulfanyl]-1,6-diazabicyclo[3.2.1]octaan6-yl-hydrosulfate;

(2S,5R)-7-oκco-2-[(τpiφtormetoκci)methyl]-1,6-diazabicyclo[3.2.1]octaan-6yl-hydrosulfate;

(2R,5R)-2-φτop-7-oκco-1,6-diazabicyclo[3.2.1]octaan-6-yl-hydrocylo;

(2S,5R)-2-φτop-7-oκco-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate; and (2R,5R)-2-chloro-7-oκco-1,6-diazabicyclo[3.2.1]octaan-6-yl-hydrocylφat;

and their pharmaceutically acceptable salts.

Preferred pharmaceutically acceptable salts of these compounds are those containing the SO ₃ H group, in particular quaternary ammonium salts, eg tetrabutylammonium salts, and alkali metal salts, eg sodium or potassium salts. Most preferred are the sodium salts, which are listed below:

- 12 042632 sodium (28.5H)-7-oxo-2-(trifluoromethyl)-1,6-diazabicyclo[3.2.1]octan-6yl sulfate;

sodium (28.5N)-7-oxo-2-(difluoromethyl)-1,6-diazabicyclo[3.2.1]octan-6yl sulfate;

sodium (28.5H)-2-(dichloromethyl)-7-oxo-1,6-diazabicyclo[3.2.1]octan-6yl sulfate;

sodium (28.5H)-2-(fluoromethyl)-7-oxo-1,6-diazabicyclo[3.2.1]octan-6yl sulfate;

sodium (28^)-2-(1,1-difluoroethyl)-7-oxo-1,6-diazabicyclo[3.2.1]octan-6yl sulfate;

sodium (28.5N)-2-[difluoro(1,3-thiazol-2-yl)methyl]-7-oxo-1,6d andazabicyclo[3.2.1]octan-6-ylsulfate; sodium (28.5H)-2-(chloromethyl)-7-oxo-1,6-diazabicyclo[3.2.1]octan-6yl sulfate; sodium (2B,5P)-7-oxo-2-[(trifluoromethyl)sulfanyl]-1,6d andazabicyclo[3.2.1]octan-6-ylsulfate; sodium (28.5N)-7-oxo-2-[(trifluoromethoxy)methyl]-1.6d andazabicyclo[3.2.1]octan-6-ylsulfate; sodium (2B,5B)-2-fluoro-7-oxo-1,6-diazabicyclo[3.2.1]octan-6-yl sulfate; sodium (28.5B)-2-fluoro-7-oxo-1,6-diazabicyclo [3.2.1]octan-6-yl sulfate; and sodium (2B,5B)-2-chloro-7-oxo-1,6-diazabicyclo[3.2.1]octan-6-yl sulfate. The most preferred compounds in which R is not halogen are (28^)-7-oxo-2-(trifluoromethyl)-1,6-diazabicyclo[3.2.1]octan-6-ylhydrosulfate;

(28^)-7-oxo-2-(difluoromethyl)-1,6-diazabicyclo[3.2.1]octan-6-ylhydrosulfate; and (28^)-2-(dichloromethyl)-7-oxo-1,6-diazabicyclo[3.2.1]octan-6-ylhydrosulfate; and their pharmaceutically acceptable salts.

Preferred pharmaceutically acceptable salts of these compounds are those containing the SO ₃ H group, in particular quaternary ammonium salts, eg tetrabutylammonium salts, and alkali metal salts, eg sodium or potassium salts. Most preferred are the sodium salts as follows: sodium (28.5H)-7-oxo-2-(trifluoromethyl)-1,6-diazabicyclo[3.2.1]octan-6ylsulfate; sodium (28.5H)-7-oxo-2-(difluoromethyl)-1,6-diazabicyclo[3.2.1]octan-6ylsulfate; and sodium (28.5H)-2-(dichloromethyl)-7-oxo-1,6-diazabicyclo[3.2.1]octan-6yl sulfate. The most preferred compounds in which R is halogen are: (2R,5R)-2-φτop-7-oκco-1,6-diazabicyclo[3.2.1]octaan-6-yl-hydrocylφatτ; (28.5H)-2-fluoro-7-oxo-1,6-diazabicyclo[3.2.1]octan-6-yl hydrogen sulfate; (2H,5H)-2-chloro-7-oxo-1,6-diazabicyclo[3.2.1]octan-6-yl hydrogen sulfate; and their pharmaceutically acceptable salts. Preferred pharmaceutically acceptable salts of these compounds are those containing the SO ₃ H group, in particular quaternary ammonium salts, eg tetrabutylammonium salts, and alkali metal salts, eg sodium or potassium salts. The most preferred are

- 13 042632 sodium salts, which are given below:

sodium (2R,5R)-2-φτop-7-oκco-1,6-diazabicyclo[3.2.1]octaan-6-ylsulfat;

sodium (2S,5R)-2-φτop-7-oκco-1,6-diazabicyclo[3.2.1]octaan-6-ylsylφat;

sodium (2R,5R)-2-chloro-7-oκco-1,6-diazabiL^iclo[3.2.1]octaan-6-ylsulfat.

Synthesis

Compounds of the invention (especially compounds of formula (I), formula (II) or formula (III) in which R is not halogen) can be synthesized according to the following scheme A.

Scheme A

1) MsOH, EtOAc

2) JISC ₃ (water)

2) oxalic acid, EtOAc, acetone

l)NaBH(OAc) ₃ , H ₂ SO ₄ , EtOAc

1) KNSO ₃ (aq.), MeTHF

2) triphosgene, NEt ₃ , MeTHF

BOC-pyrrolidinone (Boc means tert-butoxycarbonyl) (2) can be reacted with a reagent, sulfoxonium ylide, to form in situ a new open ring ylide (6). After protonation, a positively charged sulfoxonium species is formed, which is subjected to nucleophilic attack from the chloride anion, which leads to the loss of DMSO (dimethyl sulfoxide) and the formation of chloromethyl ketone, which condenses with O-benzylhydroxylamine to form oxime (7). It does not need to be isolated, but it can be treated with acid to remove the BOC protecting group, and on alkalinization spontaneous cyclization occurs with formal loss of HCl to form an extended ring (8). Again, isolation is not necessary, but stereoselective reduction of the double bond in the oxime followed by salt formation with oxalic acid facilitates the isolation of piperidine (3). Cyclization using triphosgene as a source of phosgene gives bicyclic urea (9), then after debenzylation and sulfonation, the compound of formula (I) is obtained. If necessary, the group R can be protected using suitable methods of introduction of protection. Additional details regarding synthetic routes and alternative synthesis schemes for preparing the compounds of the invention are set forth in the General Synthetic Methodology section below.

Scheme A shows routes for the synthesis of compounds of the invention of formula (II). Compounds according to the invention of formula (III) can be obtained using similar chemical transformations, but starting from the diastereoisomer (2), i. compounds (2A):

(2A)

Compounds of formula (II) and formula (III) in which R is halogen can be synthesized from the precursor, carboxylic acid, according to Scheme B below (shown for compounds of formula (II)).

- 14 042632

Scheme B

(1) AgF, Selectfluor (2) Pb(OAc)4, LiCl

The starting material in Scheme B corresponds to compound (9) in Scheme A and can be synthesized according to Scheme A starting from the carboxyl-functionalized compound (2). The halogenated compounds shown in Scheme B can be converted from the -OBn form to the -OSO form.₃ by debenzylation and sulfonation as described above and in the examples section. The corresponding compounds of formula (III) can be obtained starting from the stereoisomer of the starting material.

In order to obtain the halogen compounds of the invention, the inventors developed the chemical route shown in Scheme B. Although the process for converting aryl-containing or alkyl-containing carboxylic acids to the corresponding aryl- or alkyl-containing halides by decarboxylation has been known for many years (for example, see Hunsdiecker and Hunsdiecker (1942, Chem. Ber., 75, 291)), only in recent years has an original concept of the interaction of a carboxylic acid with a silver salt and bromine been developed. For example, Li et al. (2012, JACS, 134, 10401) reported very mild conditions for converting alkyl acids to alkyl fluorides using silver nitrate and Selectfluor™ as catalysts. Li's work includes one example of the application of the very rare functional group F-CH ₂ -N-CO, but this is a special situation where there are two carbonyl groups on the same nitrogen atom, with the effective removal of the nitrogen lone pair due to significant delocalization towards oxygen atoms carbonyl groups. There are very few examples of this functional group with only one carbonyl group on the nitrogen atom, and even these are limited to reactive intermediates for glycosylation type reactions proceeding through the formation of an iminium ion followed by quenching the iminium ion with alcohol (2016, Organic Letters, 18, 9890). The inventors have found that the tendency of a molecule to fragment and decompose can be overcome by organizing the functional group in such a way that the natural tendency to reactivity can be overcome by stereochemical control. In the case of the stability of the compounds of the invention, stabilization can be achieved through a degradation pathway by passing the iminium particles through the bridging double bond. The impossibility of forming a bridging double bond was first discovered by Bredt (1902, Ueber isomere Dehydrocamphersauren, Lauronolsauren und Bihydrolauro-Lactone, Ber. Deutsch. Chem. Ges., 35, 1286-1292), and later substantiated using the theory of molecular orbitals, according to which for formation of a double bond requires good spatial overlap of the β-orbitals. This is the case for the halogenated compounds of the invention. The introduction of a chlorine atom as a substituent is carried out in a similar manner by decarboxylation halogenation using standard Cauchy reaction conditions with lead tetraacetate and lithium chloride (JK Kochi, 1965, JACS, 87, 2500).

Compositions and combinations

The compounds of the invention may be presented in a pharmaceutical composition, wherein the pharmaceutical composition contains a compound of the invention together with a pharmaceutically acceptable carrier or diluent. Usually the composition contains up to 85 wt.% inclusive of the compounds according to the invention. In a more typical case, it contains up to 50 wt.% inclusive of the compounds according to the invention. Preferred pharmaceutical compositions are sterile and non-pyrogenic. In addition, if the pharmaceutical compositions proposed according to this invention contain a compound according to the invention, which is optically active, then this compound according to

- 15 042632 of the invention usually represents a substantially pure optical isomer.

As explained above, the compounds of the invention are useful in the treatment or prevention of bacterial infection. In particular, they are inhibitors of SBL enzymes and are therefore useful in eliminating or reducing antibiotic resistance in Gram-negative bacteria. These compounds can be used as monotherapy, or they can be used in combination therapy regimens with antibiotics to enhance the action of the antibiotic. The compounds may also be used in combination with MBL inhibitors, particularly if the bacteria causing the infection are resistant to antibiotic treatment as monotherapy and such resistance is or is implied to be caused at least in part by the presence of metallo-β-lactamase enzymes. .

Thus, the present invention also provides a combination of (1) a compound of the invention; and one or more of (2) a metallo-b-lactamase (MBL) inhibitor; and (3) an antibiotic.

If desired, the combinations may also contain other active agents.

In the combinations of the invention, each of the active agents may be present in a single composition, or one or more of them may be formulated in a separate composition. In the case of preparing separate compositions, these two or more than two agents can be administered simultaneously or separately. The active agents from which the individual compositions are prepared may be presented in the form of a kit, possibly together with instructions for their administration. For example, the kit may include (1) a composition containing a compound of the invention; and any one or both of (2) an antibiotic-containing composition and (3) an MBL inhibitor-containing composition, optionally along with instructions for their use. In the case of preparing a joint composition, these two or more than two active agents can be presented in the form of a pharmaceutical composition. Thus, the pharmaceutical composition may contain (1) the compound of the invention described in this application;

(2) a pharmaceutically acceptable carrier or diluent; and possibly one or both of (3) an antibiotic; and (4) a metallo-b-lactamase (MBL) inhibitor.

Thus, the compound of the invention and the antibiotic may be formulated together or separately. In addition, the compound of the invention and the MBL inhibitor may be formulated together or separately. If all three active agents are present, then the compound of the invention, the MBL inhibitor and the antibiotic may be presented in a single composition, or they may be formulated into separate compositions. Alternatively, two of these components may be presented in a single composition, and the remaining component may be presented separately. In other words, a composition based on a compound of the invention can be formulated together with an MBL inhibitor and an antibiotic; or a composition based on the compound of the invention may be formulated together with an MBL inhibitor while the antibiotic is presented separately; or a composition based on the compound of the invention may be formulated together with an antibiotic while the MBL inhibitor is presented separately; or the composition may be based on the MBL inhibitor together with the antibiotic, while the compound of the invention is presented separately; or based on each of the compound of the invention, the MBL inhibitor and the antibiotic, separate compositions can be prepared.

Preferably, the antibiotic administered with a compound of the invention or used in a combination or composition of the invention is a β-lactam antibiotic. More preferably, the antibiotic is a β-lactam antibiotic selected from carbapenems, penicillins, cephalosporins and penems; or the antibiotic is aztreonam. Examples of antibiotics of the carbapenem group include benapenem, imipenem, meropenem, ertapenem, doripenem and biapenem. Examples of penicillins include amoxicillin, ampicillin, ticarcillin, piperacillin and cloxacillin. Examples of cephalosporins include cefepime, cefazolin, ceftriaxone, ceftazidime, and ceftobiprole. Examples of penems include faropenem. Other antibiotics include tobramycin, neomycin, streptomycin, gentamicin, tazobactam, rifampicin, ciprofloxacin, amikacin, colistin, aztreonam, and levofloxacin. Preferably the β-lactam antibiotic is aztreonam or a carbapenem antibiotic, more preferably biapenem, imipenem or meropenem. In one aspect, the preferred carbapenem antibiotic is biapenem. In another aspect, a preferred carbapenem antibiotic is meropenem. In another preferred aspect, the β-lactam antibiotic is a carbapenem antibiotic, more preferably imipenem or meropenem, most preferably meropenem.

If an MBL inhibitor is present in the combinations or compositions of the invention or is administered together with a compound of the invention, then that MBL inhibitor is preferably a compound as described in WO 2014/198849, GB 2533136, PCT/EP 2018/069827 (which published as

WO 2019/016393) or EP 18290056.3.

Preferably the MBL inhibitor is a compound of formula (A) or a pharmaceutically acceptable salt thereof,

FORMULA (A) where

R ^1A is selected from H, R ^1b and -CH2OC(O)R ^1b where R ^1b is selected from an unsubstituted C ₁ -C ₄ alkyl group and phenyl;

® is a cyclic group selected from C ₆ -C ₁₀ aryl, 5-10 membered heteroaryl and 4-10 membered carbocyclic and heterocyclic groups;

each R ^2A is independently selected from (1) halogen or R ⁸ ;

(2) groups C ₁ . ₃ alkyl, O (C ₁ . ₃ alkyl), S (C ₁ . ₃ alkyl), SO (C ₁ . ₃ alkyl) or SO ₂ (C ₁ . ₃ alkyl), each of which may optionally be substituted by 1, 2 or 3 halo substituents and/or one R ⁸ substituent; and (3) NR ^a C(O)R ^c and NR ^a C(O)NR ^b R ^c , where R ^a and R ^b are each independently selected from hydrogen and unsubstituted C1.2 alkyl, and each R ^c is unsubstituted C1 . ₂ alkyl; and each R ⁸ is independently selected from CN, OH, -C(O)NRfRg, -NRfRg, -NR ¹⁰ C(NR ¹¹ )R ¹² , -C(NR ¹⁰ )NR ¹¹ R ¹² and -NR ¹⁰ C(NR ¹¹ )NR ¹² R ¹³ ; wherein Rf and Rg are each independently H or unsubstituted C ₁ . ₂ alkyl;

m is 0, 1, 2, or 3;

R ^3A is selected from a hydrogen atom and a C ₁ -C ₃ alkyl group which is unsubstituted or substituted with 1, 2 or 3 substituents selected from halogen, -OR ¹⁰ and -NR ¹⁰ R ¹¹ ;

n is 0 or 1;

Z ^A is a bond or is selected from

-NR ¹⁰ C(O)-, -C(O)NR ¹⁰ -, -NR ¹⁰ C(O)NR ¹¹ -, -NR ¹⁰ C(O)O-, -OC(O)NR ¹⁰ , -NR ¹⁰ C(O)S-, -SC(O)NR ¹⁰ , -NR ¹⁰ C(NR ¹¹ )-, -C(NR ¹⁰ )NR ¹¹ -, -NR ¹⁰ C(NR ¹¹ )NR ¹² -, -NR ¹⁰ C(N ⁺ R ¹¹ R ¹² )-, -C(N ⁺ R ¹⁰ R ¹¹ )NR ¹² -,

-NR ¹⁰ C(N ⁺ R ¹¹ R ¹² )NR ¹³ -, -NR ¹⁰ C(NR ¹¹ )O-, -OC(NR ¹⁰ )NR ¹¹ , -NR ¹⁰ C(N ⁺ R ¹¹ R ¹² )O- , -OC(N ⁺ R ¹⁰ R ¹¹ )NR ¹² -, -NR ¹⁰ C(NR ¹¹ )S-, -SC(NR ¹⁰ )NR ¹¹ , -NR ¹⁰ C(N ⁺ R ¹¹ R ¹² )S-, -SC(N ⁺ R ¹⁰ R ¹¹ )NR ¹² -, -C(O)NR ¹⁵ -, -NR ¹⁰ C(O)NR ¹⁵ -, -OC(O)NR ¹⁵ , -SC(O)NR ¹⁵ , -C(NR ¹⁰ )NR ¹⁵ -, -NR ¹⁰ C(NR ¹¹ )NR ¹⁵ -, -C(N ⁺ R ¹⁰ R ¹¹ )NR ¹⁵ -, -NR ¹⁰ C(N ⁺ R ¹¹ R ¹² )NR ¹⁵ -, -OC(NR ¹⁰ )NR ¹⁵ , -OC(N ⁺ R ¹⁰ R ¹¹ )NR ¹⁵ -, -SC(NR ¹⁰ )NR ¹⁵ and -SC(N ⁺ R ¹⁰ R ¹¹ )NR ¹⁵ -;

L ^a is a bond or is selected from C ₁ . ₄ alkylene, C ₂ . ₄ alkenylene, C ₂ . ₄ alkynylene, C ₁ . ₃ alkylene-(C ₃ . ₆ cycloalkylene) -C ₁ . ₃ alkylene, C ₁ . ₄ alkylene-(C _{3 .6} cycloalkylene) and (C _{3 .6} cycloalkylene)C ₁ . ₄ alkylene, wherein L is unsubstituted or substituted with 1 or 2 substituents selected from halo, -OR ¹⁰ and -NR ¹⁰ R ¹¹ ; or L is -C(R ¹⁰ )=N-;

X ^A is a bond or if L is not a bond or -C(R ¹⁰ )=N- then X is a bond or is selected from -NR ¹⁰ -, -O-, -NR ¹⁰ C(NR ¹¹ )- and -C(NR ¹⁰ )-;

p is 0 or 1;

R ^4A is selected from H, -CN and C ₁ -C ₃ alkyl which is unsubstituted or substituted with 1, 2 or 3 substituents selected from halo, -OR ¹⁰ , -NR ¹⁰ R ¹¹ and -CN;

or R ^4A is connected to R ^5A to form, together with the atoms to which they are attached, a 5-6 membered heterocyclic group containing at least one saturated carbon atom in the ring, said heterocyclic group being unsubstituted or substituted by 1 or 2 substituents, selected from unsubstituted C ₁ -C ₂ alkyl, halogen, -OR ¹⁰ , -NR ¹⁰ R ¹¹ and -CN;

R ^5A is selected from H, -CN and C ₁ -C ₃ alkyl which is unsubstituted or substituted with 1, 2 or 3 substituents selected from halo, -OR ¹⁰ , -NR ¹⁰ R ¹¹ and -CN;

or R ^5A is connected to R ^4A to form, together with the atoms to which they are attached, a 5-6 membered heterocyclic group containing at least one saturated carbon atom in

- 17 042632 ring, while the specified heterocyclic group is not substituted or substituted by 1 or 2 substituents selected from unsubstituted C1-C ₂ alkyl, halogen, -OR ¹⁰ , -NR ¹⁰ R ¹¹ and -CN;

or R ^5A is connected to R ⁶ to form, together with the atoms to which they are attached, a 5-6 membered heterocyclic group containing at least one saturated carbon atom in the ring, said heterocyclic group being unsubstituted or substituted by 1 or 2 substituents, selected from unsubstituted C1- _C2 alkyl, halogen, -OR ¹⁰ , -NR ¹⁰ R11 and -CN;

R ^6A is selected from H, -CN and C1-C ₃ alkyl which is unsubstituted or substituted with 1, 2 or 3 substituents selected from halo, -OR ¹⁰ , -NR ¹⁰ R11 and -CN;

or R ^6A is connected to R ^5A to form, together with the atoms to which they are attached, a 5-6 membered heterocyclic group containing at least one saturated carbon atom in the ring, said heterocyclic group being unsubstituted or substituted by 1 or 2 substituents, selected from unsubstituted C1- _C2 alkyl, halogen, -OR ¹⁰ , -NR ¹⁰ R11 and -CN;

or R ^6A is connected to R ^7A , if present, to form, together with the atoms to which they are attached, a 5-6 membered heterocyclic group containing at least one saturated carbon atom in the ring, said heterocyclic group being unsubstituted or substituted with 1 or 2 substituents selected from unsubstituted C1- _C2 alkyl, halo, -OR ¹⁰ , -NR ¹⁰ R11 and -CN;

R ^7A , if present, is selected from H, -CN and C1-C ₃ alkyl which is unsubstituted or substituted with 1, 2 or 3 substituents selected from halo, -OR ¹⁰ , -NR ¹⁰ R11 and -CN;

or R ^7A is connected to R ^6A to form, together with the atoms to which they are attached, a 5-6 membered heterocyclic group containing at least one saturated carbon atom in the ring, said heterocyclic group being unsubstituted or substituted by 1 or 2 substituents, selected from unsubstituted C1- _C2 alkyl, halogen, -OR ¹⁰ , -NR ¹⁰ R11 and -CN;

each R ¹⁰ , R ¹¹ , R ¹² , R ¹³ and R ¹⁴ independently represents H or methyl;

each R ¹⁵ is independently substituted C 1 -C ₄ alkyl or unsubstituted C ₂ -C ₄ alkyl, wherein if R ¹⁵ is a substituted alkyl group, then that alkyl group is substituted with 1, 2, or 3 substituents independently selected from halogen, CN, OR ¹⁰ and -NR ¹⁰ R ¹¹ .

More preferably, the MBL inhibitor is a compound of formula (A) or a pharmaceutically acceptable salt thereof, wherein

R ^1a is H;

® f is selected from phenyl, cyclohexane, piperidine, pyridazine, pyridine and thiazole;

m is 1 or 2;

each R ^2A is independently selected from halogen, CN, OH, -C(O)NRfR ^s , -NRfR ^s ; wherein each of R ^f and R ^s is independently H or methyl; and groups C ₁ . ₂ αkyl, O( _C1-2 alkyl), S(C _{1 .2} alkyl), SO(C _{1 .2} alkyl), each of which may optionally be substituted with 1, 2 or 3 substituents selected from halogen, CN, OH ; n is 0;

Z ^A is selected from -NR ¹⁰ C(O)-, -C(O)NR ¹⁰ - and -NR ¹⁰ C(O)NR ¹¹ -; L ^A is a bond or is selected from C ₁ . ₃ αkylene and C ₂ . ₃ alkenylene;

X ^A is a bond;

p is 0; or p is 1 and R ^7A is H;

R ^4A is H;

R ^5A is selected from H, -CN and C1-C ₂ alkyl which is unsubstituted or substituted with 1, 2 or 3 halo substituents and/or one -NR ¹⁰ R ¹¹ H substituent; And

R ^6A is H.

Even more preferably, the MBL inhibitor is a compound of formula (A) selected from

- 18 042632

5-[[4-[(2-guanidinoacetyl)amino]-3(trifluoromethoxy)phenyl]sulfonylamino]thiazole-4-carboxylic acid;

5-[[3-fluoro-4-[[(2-guanidinoacetyl)amino]methyl]phenyl]sulfonylamino]thiazole-4-carboxylic acid;

5-[[3-fluoro-4-(guanidinomethyl)phenyl]sulfonylamino]thiazole-4-carboxylic acid;

5-[[3-fluoro-4-(2-guanidinoethylsulfanylcarbonylamino)phenyl]sulfonylamino]thiazole-4-carboxylic acid;

5-[[4-[2-[(2-amino-2-imino-ethyl) amino]-2-oxo-ethyl]-3-fluorophenyl]sulfonylamino]thiazole-4-carboxylic acid;

5-[[3-carbamoyl-4-[(2guanidinoacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylic acid;

5-[[3-cyano-4-[(2-guanidinoacetyl)amino]phenyl]sulfonylamino]thiazole-4carboxylic acid;

5-[[3-fluoro-4-(2-guanidinoethoxycarbonylamino)phenyl]sulfonylamino]thiazole-4-carboxylic acid;

5-[(4-guanidinophenyl)sulfonylamino]thiazole-4-carboxylic acid;

5-[[4-[2-(2-carbamimidoylhydrazino)-2-oxo-ethyl]-3-fluorophenyl]sulfonylamino]thiazole-4-carboxylic acid;

5-[[3-chloro-4-[(2-guanidinoacetyl)amino]phenyl]sulfonylamino]thiazole-4carboxylic acid;

5-[[4-[(2-guanidinoacetyl)amino]-3-methoxy-phenyl]sulfonylamino]thiazole4-carboxylic acid;

5-[[4-[[2-(2-carbamimidoylhydrazino)acetyl]amino]-3-fluorophenyl]sulfonylamino]thiazole-4-carboxylic acid;

5-[[4-[[(2E)-2-(carbamimidoylhydrazono)acetyl]amino]-3-fluorophenyl]sulfonylamino]thiazole-4-carboxylic acid;

5-[[4-[[2-(4,5-dihydro-1H-imidazol-2-ylamino)acetyl]amino]-3,5-difluorophenyl]sulfonylamino]thiazole-4-carboxylic acid;

5-[[6-[(2-guanidinoacetyl)amino]pyridazin-3-yl]sulfonylamino]thiazole-4carboxylic acid;

5-[[4-[(2-Amino-2-imino-ethyl)carbamoylamino]-3-fluorophenyl]sulfonylamino]thiazole-4-carboxylic acid;

5-[[3,5-difluoro-4(guanidinocarbamoylamino)phenyl]sulfonylamino]thiazole-4-carboxylic acid;

5-[[4-[(3-Amino-3-iminopropanoyl)amino]-3,5-difluorophenyl]sulfonylamino]thiazole-4-carboxylic acid;

5-[[4-[[3-(dimethylamino)-3-iminopropanoyl]amino]-3-fluorophenyl]sulfonylamino]thiazole-4-carboxylic acid;

5-[[3-fluoro-4-[(2-guanidinooxyacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylic acid;

5-[[3-fluoro-4-[[3-imino-3-(methylamino)propanoyl]amino]phenyl]sulfonylamino]thiazole-4-carboxylic acid;

5-[[4-[3-(4,5-dihydro-1H-imidazol-2-yl)propanoylamino]-3-fluorophenyl]sulfonylamino]thiazole-4-carboxylic acid;

5-[[2-[(2-guanidinoacetyl)amino]thiazol-5-yl]sulfonylamino]thiazole-4carboxylic acid;

- 19 042632

5-[[4-[[2-[(N-cyanocarbamimidoyl)amino]acetyl]amino]-3-fluorophenyl]sulfonylamino]thiazole-4-carboxylic acid;

5-[[3-fluoro-4-(guanidinocarbamoylamino)phenyl]sulfonylamino]thiazole-4carboxylic acid;

5-[[3-fluoro-4-[[2-(morpholine-4-carboximidoylamino)acetyl]amino]phenyl]sulfonylamino]thiazole-4-carboxylic acid;

5-[[4-[(3-Amino-3-imino-2-methylpropanoyl)amino]-3-fluorophenyl]sulfonylamino]thiazole-4-carboxylic acid;

5-[[4-[[2-(4,5-dihydro-1H-imidazol-2-yl)acetyl]amino]-3-fluorophenyl]sulfonylamino]thiazole-4-carboxylic acid;

5-[[4-(carbamimidoylcarbamoylamino)-3-fluorophenyl]sulfonylamino]thiazole-4-carboxylic acid;

5-[[4-[[(2K)-2-guanidinopropanoyl]amino]phenyl]sulfonylamino]thiazole-4carboxylic acid;

5-[[3,5-difluoro-4-[(2guanidinoacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylic acid;

5-[[4-[(4-amino-4-iminobutanoyl)amino]-3-fluorophenyl]sulfonylamino]thiazole-4-carboxylic acid;

5-[[4-[[2-(4,5-dihydro-1H-imidazol-2-ylamino)acetyl]amino]-2,5-difluorophenyl]sulfonylamino]thiazole-4-carboxylic acid;

5-[[2,5-difluoro-4-[(2guanidinoacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylic acid;

5-[[3-fluoro-4-[[2-[(M-methylcarbamimidoyl)amino]acetyl]amino]phenyl]sulfonylamino]thiazole-4-carboxylic acid;

5-[[3-fluoro-4-[[2-(2-iminoimidazolidin-1yl)acetyl]amino]phenyl]sulfonylamino]thiazole-4-carboxylic acid;

5-[[4-[[2-[carbamimidoyl(methyl)amino]acetyl]amino]-3-fluorophenyl]sulfonylamino]thiazole-4-carboxylic acid;

5-[[4-[[2-[[M-(2-Aminoethyl)carbamimidoyl]amino]acetyl]amino]-3-fluorophenyl]sulfonylamino]thiazole-4-carboxylic acid;

5-[[5-fluoro-6-[(2-guanidinoacetyl)amino]-3pyridyl]sulfonylamino]thiazole-4-carboxylic acid;

5-[[3-fluoro-4-(3-guanidinopropanoylamino)phenyl]sulfonylamino]thiazole4-carboxylic acid;

5-[[4-[(3-Amino-3-iminopropanoyl)amino]-3-fluorophenyl]sulfonylamino]thiazole-4-carboxylic acid;

5-[[3,5-difluoro-4(guanidinocarbamoylamino)phenyl]sulfonylamino]thiazole-4-carboxylic acid;

5-[[3-fluoro-4-[(2-guanidinoacetyl)amino]phenyl]sulfonylamino]thiazole-4carboxylic acid; And

5-[[4-[(2-guanidinoacetyl)amino]phenyl]sulfonylamino]thiazole-4carboxylic acid;

and their pharmaceutically acceptable salts.

Compounds of formula (A) and their pharmaceutically acceptable salts are described in WO 2019/016393.

Alternatively, the MBL inhibitor may preferably be a compound of formula (B) or a pharmaceutically acceptable salt thereof,

- 20 042632

Where

R ^1B is hydrogen, halogen, CN, R ^12B , OR ^12B , SR ^12B or NR ^12B R ^13B , where R ^12B is C _1-6 alkyl optionally substituted with one or more R ^aB substituents; phenyl or 5-6 membered heteroaryl, each of which may optionally be substituted with one or more R ^bB substituents; or 3-6 membered cycloalkyl or 3-6 membered heterocyclyl, each optionally substituted with one or more R ^cB substituents;

each R ^aB is independently halogen, CN, OH or OC _1-4 alkyl optionally substituted with one or more substituents selected from halogen and OH;

each R ^bB is independently halogen, CN, OH, or C _1-4 alkyl or OC _1-4 alkyl, each of which may optionally be substituted with one or more substituents selected from halogen and OH;

each R ^cB is independently halogen, CN, OH, oxo or C _1-4 alkyl or OC _1-4 alkyl optionally substituted with one or more substituents selected from halogen and OH;

R ^13B represents a hydrogen atom or C _1-6 alkyl optionally substituted with one or more substituents R ^a as defined above;

Y ^B is a single bond, -C _1-4 alkylene- or -C _2-4 alkenylene-, each of which may be substituted with an R ^17B group; or

-C _1-4 alkylene-O-; -C _1-4 alkylene-N(R ^8B )-; -N(R ^8B )-; -C1-4alkylene-C(O)N(R ^8B )-; -C1-4alkylene-N(R ^8B )C(O) or -N(R ^8B )C _1-4 alkylene-; wherein

R ^17B is OR ^IB , NR ^IB R ^mB , NR ^IB C(O)R ^mB , C(O)NR ^IB R ^mB , C(O)OR ^mB ;

each R ^IB and RmB is independently H or C1-4αkyl; and R ^8B represents a hydrogen atom or C 1-6 alkyl or —C(O)C _1-6 alkyl, each of which is optionally substituted with one or more R ^dB substituents; And

The C _1-4 alkylene chains may optionally be substituted with one or more R ^eB substituents;

each RdB and R ^eB is independently halogen, CN, OH or OC _1-4 alkyl optionally substituted with one or more substituents selected from halogen and OH;

A ^B is a cyclic group selected from 6-10 membered aryl, 5-10 membered heteroaryl, or 3-10 membered carbocyclyl or heterocyclyl group;

nB is 0-4;

each R ^2B is independently selected from R ^3B ; or groups C _1-4 alkyl, C _2-4 alkenyl, O(C _1-4 αkyl), S(C _1-4 alkyl), SO(C _1-4 alkyl) or SO ₂ (C _1-4 alkyl) , each of which may optionally be substituted with one or more R ^3B substituents; or C(O)OR ^6B ; C(O)R ^6B ; OR ^5B , NR ^4B R ^5B ; NR ^4B C(O)R ^6B , NR ^4B C(O)NR ^5B R ^6B or SO ₂ NR ^21B R ^22B ;

or if the group A ⁱⁿ is saturated or partially saturated, then R ^2B may also be oxo;

each R ^3B is independently halogen, nitro, CN, OH; or -C(O)OR ^14B , -C(O)NR ^14B R ^15B or -NR ^14B R ^15B ; or phenyl optionally substituted with one or more R ^7B substituents; or naphthyl optionally substituted with one or more R ^7B substituents; or

5-10 membered heteroaryl optionally substituted with one or more R ^7B substituents; or

3-8-membered carbocyclyl optionally substituted with one or more R ^7B substituents; or

3-8 membered heterocyclyl optionally substituted with one or more substituents selected from oxo and R ^7B ;

each of R ^14B and R ^15B is independently H or C _1-6 alkyl optionally substituted with one or more substituents selected from halo and OH;

each R ^7B is independently halogen, CN, OH; or C1-4alkyl or OC1-4alkyl, each of which may optionally be substituted with one or more substituents selected from halo and OH; or NRjBRkB, where ^RjB and RkB are each independently H or C1-4alkyl;

each of R ^21B and R ^22B represents a hydrogen atom or C _1-4 alkyl, or R ^21B and R ^22B together with

- 21 042632 the nitrogen atom to which they are attached may form a 5- or 6-membered heterocyclic ring, optionally containing one additional heteroatom selected from N, O and S, and optionally substituted with C _1-4 alkyl or halogen;

R ^4b represents a hydrogen atom or C 1-4 alkyl optionally substituted with halogen, CN, OH, NR ^jB R ^kB ; or OC1-4alkyl which may optionally be substituted with one or more substituents selected from halo and OH; wherein each R ^jB and R ^kB is independently H or C _1-4 alkyl;

R ^5B represents a hydrogen atom, phenyl, 5-6 membered heteroaryl, 3-8 membered carbocyclyl or 3-8 membered heterocyclyl; or

C _1-6 alkyl optionally substituted with phenyl, 5-6 membered heteroaryl, 3-8 membered carbocyclyl or 3-8 membered heterocyclyl;

wherein the phenyl and heteroaryl groups are optionally substituted with one or more RfB substituents, and the carbocyclyl and heterocyclyl groups are optionally substituted with one or more R ^gB substituents, wherein each RfB is independently halogen, CN, OH, or C _1-4 alkyl or OS _1-4 alkyl, each of which may optionally be substituted with one or more substituents selected from halo and OH;

each R ^gB is independently halogen, CN, OH, oxo, or C _1-4 alkyl, or OC _1-4 alkyl optionally substituted with one or more substituents selected from halogen and OH;

R ^6B is C _1-6 alkyl optionally substituted with one or more R ^hB ; or phenyl or 5-6 membered heteroaryl, each optionally substituted with one or more R ^1B substituents;

each R ^hB is independently halogen, CN, OH, NH2, phenyl, pyridyl, COOH, or OC1-4 alkyl, optionally substituted with one or more substituents selected from halogen and OH; each R ^1B is independently halogen, CN, OH, NH2 or C1-4 alkyl or OC _1-4 alkyl, each of which may optionally be substituted with one or more substituents selected from halogen and OH;

R ^11B represents a hydrogen atom, C _1-4 alkyl optionally substituted with halogen, or benzyl optionally substituted with halogen.

More preferably, if the MBL inhibitor is a compound of formula (B) or a pharmaceutically acceptable salt thereof, then

R ^1B represents a hydrogen atom;

Y ^B is a single bond or -C _1-4 alkylene-;

^AB is a cyclic group selected from phenyl, pyridyl, pyrazolyl, thiophenyl or benzothiophenyl;

nB is 0-3; each R ^2B is independently selected from halogen or, if group A is saturated or partially saturated, then oxo; or C _1-4 alkyl; or NR ^4b R ^5B , where

R ^4b represents a hydrogen atom or C _1-4 alkyl, a

R ^5B represents a hydrogen atom; or

NR ^4b C(O)R ^6b where R ^4b is hydrogen or methyl, a

R ^6B is C _1-4 alkyl optionally substituted with one or more substituents independently selected from OH, NH2 and OC _1-4 alkyl; And

R ^11B represents a hydrogen atom.

If the MBL inhibitor is a compound of formula (B), then the MBL inhibitor is preferably selected from

- 22 042632

5-benzenesulfonamido-1,3-thiazole-4-carboxylic acid;

5-{[(3,5-dichlorophenyl)methyl]sulfonamido}-1,3-thiazole-4-carboxylic acid;

5-(2,4,6-trimethylphenylsulfonamido)thiazole-4-carboxylic acid;

5-{[3-(trifluoromethyl)phenyl]sulfonamido}-1,3-thiazole-4-carboxylic acid;

5-(phenylmethylsulfonamido)thiazole-4-carboxylic acid;

5-(3-methoxyphenylsulfonamido)thiazole-4-carboxylic acid;

5-(2-phenylethylsulfonamido)thiazole-4-carboxylic acid;

5-(thiophene-2-sulfonamido)thiazole-4-carboxylic acid;

5-(4,5-dichlorothiophene-2-sulfonamido)thiazole-4-carboxylic acid;

5-(2,5-dichlorothiophene-3-sulfonamido)thiazole-4-carboxylic acid;

5-(2-(trifluoromethyl)phenylsulfonamido)thiazole-4-carboxylic acid;

5-(4-(trifluoromethyl)phenylsulfonamido)thiazole-4-carboxylic acid;

5-(2-chloro-5-(trifluoromethyl)phenylsulfonamido)thiazole-4-carboxylic acid;

5-(3,5-bis(trifluoromethyl)phenylsulfonamido)thiazole-4-carboxylic acid;

5-({[2-(trifluoromethyl)phenyl]methyl}sulfonamido)-1,3-thiazole-4-carboxylic acid;

5-{[(2-methylphenyl)methyl]sulfonamido}-1,3-thiazole-4-carboxylic acid;

5-((2-nitrophenyl)methylsulfonamido)thiazole-4-carboxylic acid;

5-{[(2-bromophenyl)methyl]sulfonamido}-1,3-thiazole-4-carboxylic acid;

5-(5-chlorothiophene-2-sulfonamido)thiazole-4-carboxylic acid;

5-(5-phenylthiophene-2-sulfonamido)-thiazole-4-carboxylic acid;

- 23 042632

5-(thiophene-3-sulfonamido)thiazole-4-carboxylic acid;

5-(2,5-dimethylthiophene-3-sulfonamido)thiazole-4-carboxylic acid;

5-([1,1'-biphenyl]-2-ylsulfonamido)thiazol-4-carboxylic acid;

5-((2-aminophenyl)methylsulfonamido)thiazole-4-carboxylic acid;

5-((2-acetamidophenyl)methylsulfonamido)thiazole-4-carboxylic acid;

5-((2-benzamidophenyl)methylsulfonamido)thiazole-4-carboxylic acid; (E)-5-((2-styrylphenyl)methylsulfonamido)thiazole-4-carboxylic acid; (E)-5-((2-(3-(dimethylamino)-3-oxoprop-1-en-1yl)phenyl)methylsulfonamido)thiazole-4-carboxylic acid;

5-([1,1'-biphenyl]-2-ylmethylsulfonamido)-thiazole-4-carboxylic acid;

5-((2-(trifluoromethoxy)phenyl)methylsulfonamido)thiazole-4-carboxylic acid;

5-((3-(trifluoromethyl)phenyl)methylsulfonamido)thiazole-4-carboxylic acid;

5-((3-bromophenyl)methylsulfonamido)thiazole-4-carboxylic acid;

5-((3-cyanophenyl)methylsulfonamido)thiazole-4-carboxylic acid;

5-((2-chlorophenyl)methylsulfonamido)thiazole-4-carboxylic acid;

5-(4-nitrophenylsulfonamido)thiazole-4-carboxylic acid;

5-({5-[5-(trifluoromethyl)-1,2-oxazol-3-yl]thiophen-2-yl}sulfonamido)-1,3thiazole-4-carboxylic acid;

5-(1-benzothiophene-2-sulfonamido)-1,3-thiazole-4-carboxylic acid;

5-[(5-methylthiophen-2-yl)sulfonamido]-1,3-thiazole-4-carboxylic acid;

5-[(5-bromothiophen-2-yl)sulfonamido]-1,3-thiazole-4-carboxylic acid;

5-(1-benzothiophene-3-sulfonamido)-1,3-thiazole-4-carboxylic acid;

5-[(4-bromo-2,5-dichlorothiophen-3-yl)sulfonamido]-1,3-thiazole-4-carboxylic acid;

5-({[(2-chlorophenyl)methyl]sulfamoyl}amino)-1,3-thiazole-4-carboxylic acid;

5-[({[3-(trifluoromethyl)phenyl]methyl}sulfamoyl)amino]-1,3-thiazole-4carboxylic acid;

5-[(3-bromothiophen-2-yl)sulfonamido]-1,3-thiazole-4-carboxylic acid;

5-{[(2-iodophenyl)methyl]sulfonamido}-1,3-thiazole-4-carboxylic acid; 5-{[4-phenyl-5-(trifluoromethyl)thiophen-3-yl]sulfonamido}-1,3-thiazole-4carboxylic acid;

- 24 042632

5-{[(2,3-dichlorophenyl)methyl]sulfonamido}-1,3-thiazole-4-carboxylic acid;

5-{[(3,4-dichlorophenyl)methyl]sulfonamido}-1,3-thiazole-4-carboxylic acid;

5-benzylsulfonamido-2-methyl-1,3-thiazole-4-carboxylic acid;

2-methyl-5-(quinoline-8-sulfonamido)-1,3-thiazole-4-carboxylic acid;

5-benzenesulfonamido-2-methyl-1,3-thiazole-4-carboxylic acid;

5-{[(3,5-dichlorophenyl)methyl]sulfonamido}-2-methyl-1,3-thiazole-4carboxylic acid;

5-[(2-chlorophenyl)sulfonamido]-2-methyl-1,3-thiazole-4-carboxylic acid;

2-methyl-5-[(2,4,6-trimethylphenyl)sulfonamido]-1,3-thiazole-4-carboxylic acid;

5-[(2,5-dichlorothiophen-3-yl)sulfonamido]-2-methyl-1,3-thiazole-4-carboxylic acid;

5-{[(2-bromophenyl)methyl]sulfonamido}-2-methyl-1,3-thiazole-4-carboxylic acid;

5-benzenesulfonamido-2-phenyl-1,3-thiazole-4-carboxylic acid;

5-benzenesulfonamido-2-ethyl-1,3-thiazole-4-carboxylic acid;

5-[(1-phenylethyl)sulfonamido]-1,3-thiazole-4-carboxylic acid;

5-{[6-(trifluoromethyl)pyridin-3-yl]sulfonamido}-1,3-thiazole-4-carboxylic acid;

5-[(2-phenoxyethyl)sulfonamido]-1,3-thiazole-4-carboxylic acid;

5-{[2-(1,3-dioxo-2,3-dihydro-1H-isoindol-2-yl)ethyl]sulfonamido}-1,3thiazole-4-carboxylic acid;

5-{[2-(2-chlorophenyl)ethyl]sulfonamido}-1,3-thiazole-4-carboxylic acid;

5-({1-[5-(trifluoromethyl)pyridin-2-yl]-1H-pyrazol-4-yl}sulfonamido)-1,3thiazole-4-carboxylic acid;

5-[(2-chlorophenyl)sulfonamido]-1,3-thiazole-4-carboxylic acid;

5-(pyridine-3-sulfonamido)-1,3-thiazole-4-carboxylic acid;

5-[(2,6-dichlorophenyl)sulfonamido]-1,3-thiazole-4-carboxylic acid;

5-(cyclohexylmethyl)sulfonamido-1,3-thiazole-4-carboxylic acid;

5-[(4-methyl-3,4-dihydro-2H-1,4-benzoxazin-7-yl)sulfonamido]-1,3-thiazole4-carboxylic acid;

5-[(1-phenylpropyl)sulfonamido]-1,3-thiazole-4-carboxylic acid;

- 25 042632

5-{[2-(4-methoxyphenyl)ethyl]sulfonamido}-1,3-thiazole-4-carboxylic acid;

5-({2-[3-(trifluoromethyl)phenyl]ethyl}sulfonamido)-1,3-thiazole-4-carboxylic acid;

5-{[2-(4-chlorophenyl)ethyl]sulfonamido}-1,3-thiazole-4-carboxylic acid;

5-[(piperidine-1-sulfonyl)amino]-1,3-thiazole-4-carboxylic acid;

5-[(phenylsulfamoyl)amino]-1,3-thiazole-4-carboxylic acid;

5-{[benzyl(methyl)sulfamoyl]amino}-1,3-thiazole-4-carboxylic acid;

5-[(4-acetamidophenyl)sulfonamido]-1,3-thiazole-4-carboxylic acid;

5-[(2-methoxyphenyl)sulfonamido]-1,3-thiazole-4-carboxylic acid;

5-(1,2,3,4-tetrahydronaphthalene-1-sulfonamido)-1,3-thiazole-4-carboxylic acid;

5-[(5-methyl-1-phenyl-1H-pyrazol-4-yl)sulfonamido]-1,3-thiazole-4-carboxylic acid;

5-(cyclopropylmethyl)sulfonamido-1,3-thiazole-4-carboxylic acid;

5-{[(2-methoxyphenyl)methyl]sulfonamido}-1,3-thiazole-4-carboxylic acid;

5-{[2-(2-methoxyphenyl)ethyl]sulfonamido}-1,3-thiazole-4-carboxylic acid;

5-{[2-(3-methoxyphenyl)ethyl]sulfonamido}-1,3-thiazole-4-carboxylic acid;

5-{[2-(3-chlorophenyl)ethyl]sulfonamido}-1,3-thiazole-4-carboxylic acid;

5-[(2-methanesulfonylphenyl)sulfonamido]-1,3-thiazole-4-carboxylic acid;

5-{[methyl(phenyl)sulfamoyl]amino}-1,3-thiazole-4-carboxylic acid;

5-{[4-(morpholin-4-yl)phenyl]sulfonamido}-1,3-thiazole-4-carboxylic acid;

5-[(4-cyanophenyl)sulfonamido]-1,3-thiazole-4-carboxylic acid;

5-(pyridine-2-sulfonamido)-1,3-thiazole-4-carboxylic acid;

5-[(1-methyl-1H-imidazol-2-yl)sulfonamido]-1,3-thiazole-4-carboxylic acid;

5-[(6-methoxypyridin-3-yl)sulfonamido]-1,3-thiazole-4-carboxylic acid;

5-{[4-(1H-pyrazol-1-yl)phenyl]sulfonamido}-1,3-thiazole-4-carboxylic acid;

- 26 042632

5-[(1-ethyl-5-methyl-1H-pyrazol-4-yl)sulfonamido]-1,3-thiazole-4-carboxylic acid;

5-{[(2-chlorophenyl)methyl]sulfonamido}-2-(trifluoromethyl)-1,3-thiazole-4carboxylic acid;

5-{[(2-cyanophenyl)methyl]sulfonamido}-1,3-thiazole-4-carboxylic acid;

5-[(1-methyl-1H-pyrazol-3-yl)sulfonamido]-1,3-thiazole-4-carboxylic acid;

5-[(1-methyl-1H-pyrazol-5-yl)sulfonamido]-1,3-thiazole-4-carboxylic acid;

5-({1-[(benzyloxy)carbonyl]piperidin-4-yl}sulfonamido)-1,3-thiazole-4carboxylic acid;

5-[(3-phenylpropyl)sulfonamido]-1,3-thiazole-4-carboxylic acid;

5-{[(2-chlorophenyl)methyl]sulfonamido}-2-methyl-1,3-thiazole-4-carboxylic acid;

5-(2,3-dihydro-1H-indene-1-sulfonamido)-1,3-thiazole-4-carboxylic acid;

5-{[4-(1-methyl-1H-pyrazol-5-yl)phenyl]sulfonamido}-1,3-thiazole-4carboxylic acid;

5-{[2-(1,2,3,4-tetrahydro-roquinolin-1-yl)ethyl]sulfonamido}-1,3-thiazole-4-carboxylic acid;

5-{[2-(M-phenylacetamido)ethyl]sulfonamido}-1,3-thiazole-4-carboxylic acid;

5-{[4-(3-oxomorpholin-4-yl)phenyl]sulfonamido}-1,3-thiazole-4-carboxylic acid;

5-{[4-(2-oxo-1,3-oxazolidin-3-yl)phenyl]sulfonamido}-1,3-thiazole-4carboxylic acid;

5-[(1,2-dimethyl-1H-imidazol-4-yl)sulfonamido]-1,3-thiazole-4-carboxylic acid;

5-[(oxan-4-ylmethyl)sulfonamido]-1,3-thiazole-4-carboxylic acid;

5-[({1-[(benzyloxy)carbonyl]piperidin-4-yl}methyl)sulfonamido]-1,3thiazole-4-carboxylic acid;

5-{[4-(2-oxopyrrolidin-1-yl)phenyl]sulfonamido}-1,3-thiazole-4carboxylic acid;

5-({1-[6-(trifluoromethyl)pyridin-3-yl]-1H-pyrazol-4-yl}sulfonamido)-1,3thiazole-4-carboxylic acid;

- 27 042632

5-{[4-(1,3-oxazol-5-yl)phenyl]sulfonamido}-1,3-thiazole-4-carboxylic acid;

5-{[4-(1H-pyrazol-4-yl)phenyl]sulfonamido}-1,3-thiazole-4-carboxylic acid;

5-[(1-phenyl-1H-pyrazol-4-yl)sulfonamido]-1,3-thiazole-4-carboxylic acid;

5-{[4-(piperidin-4-yl)phenyl]sulfonamido}-1,3-thiazole-4-carboxylic acid;

5-[(4-propanamidophenyl)sulfonamido]-1,3-thiazole-4-carboxylic acid;

5-{[4-(2-hydroxyacetamido)phenyl]sulfonamido}-1,3-thiazole-4-carboxylic acid;

5-({4-[(methylcarbamoyl)amino]phenyl}sulfonamido)-1,3-thiazole-4carboxylic acid;

5-{[(2,4-dichlorophenyl)methyl]sulfonamido}-1,3-thiazole-4-carboxylic acid;

5-{[(2-fluorophenyl)methyl]sulfonamido}-1,3-thiazole-4-carboxylic acid;

5-{[(2,3-difluorophenyl)methyl]sulfonamido}-1,3-thiazole-4-carboxylic acid;

5-{[4-(2-methoxyacetamido)phenyl]sulfonamido}-1,3-thiazole-4-carboxylic acid;

5-{[(2,5-dichlorophenyl)methyl]sulfonamido}-1,3-thiazole-4-carboxylic acid;

5-{[(2,6-dichlorophenyl)methyl]sulfonamido}-1,3-thiazole-4-carboxylic acid;

5-{[(2-chloro-6-fluorophenyl)methyl]sulfonamido}-1,3-thiazole-4-carboxylic acid;

5-{[(2-chloro-4-fluorophenyl)methyl]sulfonamido}-1,3-thiazole-4-carboxylic acid;

5-({[2-chloro-5-(trifluoromethyl)phenyl]methyl}sulfonamido)-1,3-thiazole-4carboxylic acid;

5-({4-[(dimethylamino)methyl]phenyl}sulfonamido)-1,3-thiazole-4-carboxylic acid;

5-{[(2,3,5-trichlorophenyl)methyl]sulfonamido}-1,3-thiazole-4-carboxylic acid;

- 28 042632

5-{[(2,3-dichloro-6-fluorophenyl)methyl]sulfonamido}-1,3-thiazole-4carboxylic acid;

5-({[2,3-dichloro-6-(trifluoromethyl)phenyl]methyl}sulfonamido)-1,3-thiazole-4carboxylic acid;

5-{[(4-bromo-2-chlorophenyl)methyl]sulfonamido}-1,3-thiazole-4-carboxylic acid;

5-({2-[methyl(phenyl)amino]ethyl}sulfonamido)-1,3-thiazole-4-carboxylic acid;

5-{[(4-nitrophenyl)methyl]sulfonamido}-1,3-thiazole-4-carboxylic acid;

5-[6-(piperidin-1-yl)pyridin-3-ylsulfonamido]-1,3-thiazole-4-carboxylic acid;

5-[6-(methylamino)pyridin-3-ylsulfonamido]-1,3-thiazole-4-carboxylic acid;

5-[6-(4-methylpiperazin-1-yl)pyridin-3-ylsulfonamido]-1,3-thiazole-4-carboxylic acid;

5-(6-acetamidopyridin-3-ylsulfonamido)-1,3-thiazole-4-carboxylic acid;

5-{4-[(5-methyl-1,2-oxazol-3-yl)amino]phenylsulfonamido}-1,3-thiazole-4carboxylic acid;

5-(6-aminopyridin-3-ylsulfonamido)-1,3-thiazole-4-carboxylic acid;

5-[(6-chloro-2H-1,3-benzodioxol-5-yl)methylsulfonamido]-1,3-thiazole-4-carboxylic acid;

5-{[(2-chloro-6-nitrophenyl)methyl]sulfonamido}-1,3-thiazole-4-carboxylic acid;

5-(quinoline-6-sulfonamido)-1,3-thiazole-4-carboxylic acid;

5-[(2,3-dihydroindole-1-sulfonyl)amino]-1,3-thiazole-4-carboxylic acid;

5-(4-methanesulfonylphenylsulfonamido)-1,3-thiazole-4-carboxylic acid;

5-[3-(2-oxo-1,3-oxazolidin-3-yl)phenylsulfonamido]-1,3-thiazole-4-carboxylic acid;

5-[3-(2H-pyrazol-3-yl)phenylsulfonamido]-1,3-thiazole-4-carboxylic acid;

5-[2-(pyridin-3-yl)ethylsulfonamido]-1,3-thiazole-4-carboxylic acid;

5-[3-(3-oxomorpholin-4-yl)phenylsulfonamido]-1,3-thiazole-4-carboxylic acid;

- 29 042632

5-[3-(2-oxopyrrolidin-1-yl)phenylsulfonamido]-1,3-thiazole-4-carboxylic acid;

5-[6-(piperidin-4-ylamino)pyridin-3-ylsulfonamido]-1,3-thiazole-4-carboxylic acid;

5-(6-{[2-(dimethylamino)ethyl]amino}pyridin-3-ylsulfonamido)-1,3-thiazole4-carboxylic acid;

5-[(4-acetamidophenyl)methylsulfonamido]-1,3-thiazole-4-carboxylic acid;

5-[6-(piperazin-1-yl)pyridin-3-ylsulfonamido]-1,3-thiazole-4-carboxylic acid;

5-[6-(4-aminopiperidin-1-yl)pyridin-3-ylsulfonamido]-1,3-thiazole-4-carboxylic acid;

5-[6-(3-Aminopyrrolidin-1-yl)pyridin-3-ylsulfonamido]-1,3-thiazole-4-carboxylic acid;

5-[6-(pyrrolidin-1-yl)pyridin-3-ylsulfonamido]-1,3-thiazole-4-carboxylic acid;

5-[6-(3-aminopiperidin-1-yl)pyridin-3-ylsulfonamido]-1,3-thiazole-4-carboxylic acid;

5-(6-(1,4-diazepan-1-yl)pyridin-3-ylsulfonamido]-1,3-thiazole-4-carboxylic acid;

5-[4-(pyrrole idin-3-yloxy)phenylsulfonamido]-1,3-thiazole-4-carboxylic acid;

5-[6-(3-aminoazetidin-1-yl)pyridin-3-ylsulfonamido]-1,3-thiazole-4-carboxylic acid;

5-[6-(piperidin-4-yl)pyridin-3-ylsulfonamido]-1,3-thiazole-4-carboxylic acid;

5-(6-(1,2,3,6-tetrahydropyridin-4-yl)pyridin-3-ylsulfonamido]-1,3thiazole-4-carboxylic acid;

5-{6-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]pyridin-3-ylsulfonamido}-1,3thiazole-4-carboxylic acid;

5-[1-(2-chlorophenyl)ethylsulfonylamino]thiazole-4-carboxylic acid;

5-[1-(2-chlorophenyl)ethylsulfonylamino]thiazole-4-carboxylic acid;

5-(3-pyridylmethylsulfonylamino)thiazole-4-carboxylic acid;

5-(isoindolin-5-ylmethylsulfonylamino)thiazole-4-carboxylic acid;

- 30 042632

C-5-[[4-[1-(2-Amino-2-phenylacetyl)-4piperidyl]phenyl]sulfonylamino]thiazole-4-carboxylic acid;

5-5-[[4-[1-(2-amino-2-phenylacetyl)-4-piperidyl]phenyl]sulfonylamino]thiazole-4-carboxylic acid;

5-[[4-[(2-aminoacetyl)amino]phenyl1sulfonylamino]thiazole-4-carboxylic acid;

5-[(4-acetamido-3-fluoro-phenyl)sulfonylamino]thiazole-4-carboxylic acid;

5-[[4-[(2-Hydroxy-2-methylpropanoyl)amino]phenyl]sulfonylamino]thiazole4-carboxylic acid;

5-[[4-[(2-Hydroxy-2-phenylacetyl)amino]phenyl]sulfonylamino]thiazole-4carboxylic acid;

5-[[4-[(2-hydroxy-3-phenylpropanoyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylic acid;

5-[[2-(2-hydroxyethylamino)pyrimidin-5-yl]sulfonylamino]thiazole-4carboxylic acid;

5-[(2-methylpyrimidin-5-yl)sulfonylamino]thiazole-4-carboxylic acid;

5-[[2-(4-pyridyl)pyrimidin-5-yl]sulfonylamino]thiazole-4-carboxylic acid;

5-[(6-methyl-3-pyridyl)sulfonylamino]thiazole-4-carboxylic acid;

5-[(2-chloro-3-nitro-phenyl)methylsulfonylamino]thiazole-4-carboxylic acid;

and their pharmaceutically acceptable salts.

Compounds of formula (B) and their pharmaceutically acceptable salts are described in WO 2014/198849.

The invention thus provides a combination of a compound of the invention as described herein, for example a compound of formula (I), (II) or (III), wherein R is as defined herein, preferably where R is is a halogen, and an MBL inhibitor of formula (A) or formula (B) as defined herein, wherein the MBL inhibitor is preferably of formula (A). The combination may additionally contain an antibiotic, which is described in this application. The combination may be in the form of a pharmaceutical composition further comprising a pharmaceutically acceptable carrier or diluent.

The compounds, compositions or combinations described in this application can be administered in a variety of dosage forms. Thus, they can be administered orally, for example, in the form of tablets, lozenges, lozenges, aqueous or oily suspensions, dispersible powders or granules. They can also be administered parenterally, whether this administration is subcutaneous, intravenous, intramuscular, intrasternal, transdermal or using infusion methods. The compound, composition or combination can also be administered as a suppository. Preferably, the compound, composition or combination is administered via parenteral administration, in particular via intravenous (IV) administration.

Compounds, compositions or combinations described in this application, usually prepared for administration together with a pharmaceutically acceptable carrier or diluent. For example, solid oral forms may contain, together with the active compound, diluents such as lactose, dextrose, sucrose, cellulose, corn starch or potato starch; lubricants, for example silica, talc, stearic acid, magnesium or calcium stearate and/or polyethylene glycols; binders; for example starches, gum arabic, gelatin, methylcellulose, carboxymethylcellulose or polyvinylpyrrolidone; disaggregating agents, for example starch, alginic acid, alginates or sodium starch glycolate; gas-evolving mixtures; coloring agents; sweeteners; wetting agents such as lecithin, polysorbates, lauryl sulfates; and generally non-toxic and pharmacologically inactive substances used in pharmaceutical compositions. Such pharmaceutical preparations may be prepared in a manner known per se, for example using blending, granulating, tableting, sugar coating or film coating processes.

Liquid dispersions for oral administration may be syrups, emulsions and suspensions. Syrups may contain as carriers, for example, sucrose or sucrose with glycerol and/or mannitol and/or sorbitol.

Suspensions and emulsions may contain as a carrier, for example, natural gum, agar,

- 31 042632 sodium alginate, pectin, methylcellulose, carboxymethylcellulose or polyvinyl alcohol. Suspensions or solutions for intramuscular injection may contain, together with the active compound, a pharmaceutically acceptable carrier such as sterile water, olive oil, ethyl oleate, glycols such as propylene glycol and, if desired, a suitable amount of lidocaine hydrochloride.

Injectable solutions may contain, for example, sterile water as a carrier, or they may preferably be in the form of sterile aqueous isotonic saline solutions. Compositions for injection (eg, for intravenous administration) may contain excipients to increase the solubility of the constituent compounds (eg, compounds of the invention). Suitable excipients include cyclodextrins such as Captisol. Pharmaceutical compositions suitable for delivery by needleless injection, eg transdermally, may also be used.

Therapeutic efficacy

The compounds of the present invention are therapeutically useful. Therefore, according to the present invention, the compounds, compositions and combinations described in this application are provided for use in medicine. The present invention provides the compounds described in this application for use in the treatment of a human or animal body. For the avoidance of doubt, the compound of the invention may be administered in the form of a solvate.

The compounds, compositions and combinations of the invention are useful in the treatment or prevention of a bacterial infection. Thus, the present invention provides a compound, composition or combination of the invention for use in the treatment or prevention of a bacterial infection. The invention also provides the use of a compound, composition or combination of the invention in the manufacture of a medicament for use in the prevention or treatment of a bacterial infection. The invention also provides a method for treating or preventing a bacterial infection in a subject in need thereof, which method comprises administering to said subject an effective amount of a compound, composition or combination described herein.

The compounds of the invention can be used as independent therapeutic agents. For example, the compounds of the invention can be used as stand-alone supplements in antibiotic therapy, for example, in chemotherapy regimens. Alternatively, they can be used in combination with antibiotics and possibly MBL inhibitors to enhance the action of the antibiotic. The compounds of the invention may find particular use in the treatment or prevention of a bacterial infection caused by bacteria that are resistant to antibiotic treatment when used as monotherapy, in particular when resistance is caused by the presence of SBL enzymes. Treatment or prevention of such an infection with β-lactam antibiotics alone may not be successful. Thus, these compounds are useful in eliminating or reducing antibiotic resistance, in particular in Gram-negative bacteria. In particular, these compounds are useful in eliminating or reducing resistance caused by the presence of SBL enzymes.

The invention also provides a compound of the invention for use in the treatment or prevention of a bacterial infection by co-administration with (1) an antibiotic and/or (2) an MBL inhibitor. Also provided is an antibiotic for use in the treatment or prevention of a bacterial infection by co-administration with (1) a compound of the invention and optionally (2) an MBL inhibitor. Also provided is an MBL inhibitor for use in the treatment or prevention of a bacterial infection by co-administration with (1) a compound of the invention and possibly (2) an antibiotic.

Also provided is the use of a compound of the invention in the manufacture of a medicament for use in the treatment or prevention of a bacterial infection by co-administration with (1) an antibiotic and/or (2) an MBL inhibitor. Also provided is the use of an antibiotic in the manufacture of a medicament for use in the treatment or prevention of a bacterial infection by co-administration with (1) a compound of the invention and optionally (2) an MBL inhibitor. Also provided is the use of an MBL inhibitor in the manufacture of a medicament for use in the treatment or prevention of a bacterial infection by co-administration with (1) a compound of the invention and possibly (2) an antibiotic.

Also provided is a method for treating or preventing a bacterial infection in a subject in need thereof, the method comprising co-administration of a compound of the invention (1) and an antibiotic and/or (2) an MBL inhibitor.

In one aspect, the subject to be treated is a mammal, in particular a human. However, he may not be human.

Preferred non-human animals include, but are not limited to, primates such as marmosets or marmosets raised for sale, animals such as horses, cattle, sheep or pigs, and pets such as dogs, cats, mice, rats , guinea pigs, ferrets, gerbils or hamsters. The subject can be any animal that can

- 32 042632 become infected with bacteria.

Compounds, compositions and combinations described in this application are useful in the treatment of bacterial infection that occurs after a relapse at the end of antibiotic treatment. Thus, these compounds, compositions and combinations can be used to treat a patient previously treated with an antibiotic for (same episode of) a bacterial infection.

The bacterium causing the infection can be any bacterium that expresses the SBL enzyme or equivalent. Usually, the bacterium causing the infection expresses the SBL enzyme. Typically, the bacterium is a Gram-negative bacterium. In particular, the bacterium may be a pathogenic bacterium. Typically, a bacterial infection to be treated using the compounds of the invention is resistant to conventional antibiotic treatment when the conventional antibiotic is used as monotherapy or in combination with another partner. For example, a bacterial infection to be treated using the compounds of the invention may be resistant to treatment when a conventional antibiotic is used solely in combination with an MBL inhibitor.

Gram-negative bacteria whose antibiotic resistance can be eliminated using the compounds of general formula (I) are serine β-lactamase producing bacteria, which may be serine β-lactamases of subclasses A, C or D. In particular, bacteria can be bacteria producing extended spectrum β-lactamases (ESBL) and/or carbapenemase, in particular OXA and/or KPC class carbapenemase, preferably OXA class carbapenemase.

The bacterial infection may be caused by bacteria from the Enterobacteriaceae, Pseudomonas and/or Moraxella families, more typically the bacterial infection is caused by bacteria from the Enterobacteriaceae and/or Pseudomonas families, and most typically the bacterial infection is caused by bacteria from the Enterobacteriaceae family. A bacterial infection can be caused by Pseudomonas (eg, Pseudomonas aeruginosa, Pseudomonas oryzihabitans, or Pseudomonas plecoglossicida), Klebsiella, Escherichia, Enterobacter, Acinetobacter, or Burkholderia. Preferably, the bacterial infection may be caused by Pseudomonas (eg Pseudomonas aeruginosa, Pseudomonas oryzihabitans or Pseudomonas plecoglossicida), Klebsiella, Escherichia, Enterobacter or Acinetobacter. More preferably, the bacterial infection is caused by Acinetobacter or Klebsiella, most preferably Acinetobacter. For example, a bacterial infection can be caused by Klebsiella pneumoniae, Escherichia coli, Enterobacter cloacae, Pseudomonas aeruginosa, Burkholderia cepacia, or Acinetobacter baumannii. Preferably, the bacterial infection may be due to Klebsiella pneumoniae, Escherichia coli, Enterobacter cloacae, Pseudomonas aeruginosa or Acinetobacter baumannii. More preferably, the bacterial infection is caused by Acinetobacter baumannii or Klebsiella pneumoniae, most preferably Acinetobacter baumannii. The bacterium may be an opportunistic pathogen.

A compound, composition or combination of the invention may be used to treat or prevent infections and conditions caused by any of the aforementioned bacteria or a combination thereof. In particular, a compound, composition or combination of the invention may be used to treat or prevent pneumonia. The compound, composition, or combination may also be used to treat septic shock, urinary tract infections, and infections of the gastrointestinal tract, skin, or soft tissue.

The compound, composition or combination of the invention can be used to treat patients with carbapenem resistant enterobacteriaceae (CRE). CREs can be found in densely populated settings or in hospitals and other settings that are commonly associated with long-term patients and patients who undergo significant medical interventions, such as those who are typically treated in intensive care units ( ICU, from Intensive Care Units).

A compound, composition or combination of the invention may be administered to a subject to prevent the onset or recurrence of one or more of the symptoms of a bacterial infection. This is called prevention. In this embodiment, the infection in the subject may be asymptomatic. Typically, the subject is a subject that is exposed to the bacterium. Such a subject is administered a prophylactically effective amount of the agent or composition. A prophylactically effective amount is an amount that prevents the onset of one or more symptoms of a bacterial infection.

A compound, composition or combination of the invention may be administered to a subject for the purpose of treating one or more symptoms of a bacterial infection. In this embodiment, the subject typically exhibits symptoms. Such a subject is administered a therapeutically effective amount of the agent or composition. A therapeutically effective amount is an amount effective to reduce the intensity of one or more symptoms of a given disorder.

The subject is administered a therapeutically or prophylactically effective amount of the compound

- 33 042632 of the invention. The dose may be determined according to various parameters, in particular according to the compound used; age, weight and condition of the subject to be treated; route of administration; and the required treatment regimen. The physician will be able to determine the appropriate route of administration and dosage for any particular subject. A typical daily dose is from about 0.01 to 100 mg per kg, preferably from about 0.1 to 50 mg/kg, for example, from about 1 to 10 mg/kg of body weight, according to the specific inhibitor activity, age, weight and the condition of the subject being treated, the type and severity of the disease, and the frequency and route of administration. Preferably, daily dosage levels are from 70 mg to 3.5 g. Sometimes higher dosages are needed, such as from about 0.01 to about 250 mg per kg, for example, from about 0.1 mg/kg to about 200 mg /kg, for example, from about 1 to about 150 mg/kg of body weight, according to the activity of the particular inhibitor, the age, weight and condition of the subject to be treated, the type and severity of the disease and the frequency and route of administration. Such preferred daily dosage levels may be, for example, from 70 mg to 8 g. Higher dosages may be particularly acceptable if the compound of the invention is administered to the subject several times a day, such as 2, 3 or 4 times a day, for example, 4 times a day. A suitable daily dosage may be from 70 mg to 8 g per day administered in 2, 3 or 4 separate dosages.

If a compound of the invention is administered to a subject in combination with another active agent (e.g., in the form of a pharmaceutical combination containing an antibiotic and possibly an MBL inhibitor), then the dose of the other active agent (e.g., MBL inhibitor and/or antibiotic) may be determined as described. higher. The dose may be determined according to various parameters, in particular according to the agent used; age, weight and condition of the subject to be treated; route of administration; and the required treatment regimen. Again, the physician will be able to determine the appropriate route of administration and dosage for any particular subject. A typical daily dose is from about 0.01 to 100 mg per kg, preferably from about 0.1 to 50 mg/kg, for example, from about 1 to 10 mg/kg of body weight, according to the specific inhibitor activity, age, weight and the condition of the subject being treated, the type and severity of the disease, and the frequency and route of administration. Preferably, daily dosage levels are from 70 mg to 3.5 g. Sometimes higher dosages are needed, such as from about 0.01 to about 250 mg per kg, for example, from about 0.1 mg/kg to about 200 mg /kg, for example, from about 1 to about 150 mg/kg of body weight, according to the activity of the particular inhibitor, the age, weight and condition of the subject to be treated, the type and severity of the disease and the frequency and route of administration. Such preferred daily dosage levels may be, for example, from 70 mg to 8 g. Higher dosages may be particularly acceptable if the combination or composition of the invention is administered to the subject several times a day, such as 2, 3 or 4 times a day, for example, 4 times a day. A suitable daily dosage may be from 70 mg to 8 g per day administered in 2, 3 or 4 separate dosages.

The following examples illustrate the invention. However, they do not limit the invention in any way. In this regard, it is important to understand that the particular assay used in the Examples section is only intended to provide an indication of biological activity. There are many assays used to determine biological activity and therefore a negative result in any one particular assay is not definitive.

- 34 042632

Experimental details General synthesis methodology

1) MsOH, LiCl, THF

Me ₃ SO ⁺ r, NaH

DMSO, THF

O

Scheme 1

2) BnONH ₂ .HCI, NaOAc (water), EtOAc

1) MsOH, EtOAc

2) KNSOz (water)

1) NaBH(OAc) ₃ , H ₂ SO ₄ , EtOAc

2) Oxalic acid, EtOAc, acetone

fTrans.Cis (75:25)

1) KNSO ₃ (aq.), MeTHF

2) Triphosgene, NEt ₃ , MeTHF

1) Acetone, LiOH (aq.)

2) Pivaloyl chloride, NEt ₃ , DCM j

3) Ammonia...

35%

TranaCis (75:25) ^ ^t R ^ans ( ^100% )

H ₂ , Pd/C DMF, DCM □MF, DCM o

n

He

SO ₃ , DMF, TVAA, DXM

55%

.o o

Sodium ethyl 2-hexanoate

90%

VISCH

(10) (5) (1)

There are a number of approaches to the synthesis of avibactam (1). One of the main routes is shown in Scheme 1, reprinted from Ball M. et al., Organic Process Research and Development, 2016, 20, 1799. open loop (6). After protonation, a positively charged sulfoxonium species is formed, which is subjected to nucleophilic attack from the chloride anion, which leads to the loss of DMSO and the formation of chloromethyl ketone, which condenses with O-benzylhydroxylamine to form oxime (7). Again, it does not need to be isolated, but it can be treated with acid to remove the BOC protecting group, and on alkalization spontaneous cyclization occurs with formal loss of HCl to form an expanded ring (8). Again, it is not isolated, but the stereoselective reduction of the double bond in the oxime followed by the formation of a salt with oxalic acid facilitates the isolation of piperidine (3). Cyclization using triphosgene as a source of phosgene produces bicyclic urea (9), then after hydrolysis of the ester and formation of the amide, the primary avibactam carboxamide is obtained, as in (4). At the end, after debenzylation and sulfonation, avibactam (1) is obtained.

By using substituents other than the ester in the parent pyrrolidinone, alternative analogs of avibactam can be obtained. In addition, intermediate (3) is currently commercially available (for example, from Shanghai Habo Chemical Technology Company or Frapps Chemicals Co, Zhejiang, China, http://www.frappschem.com), so at this later intermediate step many substituents can be used. Examples are shown below in schemes 2a and b.

- 35 042632

Scheme 2a

Scheme 2b

In Schemes 2a and 2b, Z is a suitable ester substituent such as the benzyl group in compound (9) in Scheme 1. Y is a group:

where R ^pg represents a protecting group, for example, a benzyl group or a CH ₂ CH=CH ₂ group. This protecting group can be removed after the rest of the compound has been synthesized to give the SO ₃ H or SO ₃ group ^- similar to how it is shown in Scheme 1 above.

As depicted in Scheme 2a, an ester functional group such as compound (9) in Scheme 1 can be reduced to give an alcohol (11) which can be oxidized to an aldehyde

- 36 042632 (12) . The aldehyde group can be converted to difluoromethyl substituent (13) or dichloromethyl substituent (14) using DAST (diethylaminosulfur trifluoride; see Xu, Y., Prestwich, GD, Journal of Organic Chemistry, 2002, 67, 20, 7158-7161) or PCl ₅ (Gauthier, J. et al., Bioorganic and Medicinal Chemistry Letters, 2008, 18, 923-928) or similar reagents whose chemical transformations are well known to those skilled in the art. Similarly, the alcohol group can be converted to fluoromethyl (15) using DAST (Liu, Y. et al., Organic Letters, 2004, 6, 209-212) and chloromethyl (16) using thionyl chloride (Gudipati, V. et al. , Journal of Organic Chemistry, 2006, 71, 3599-3607). In addition, using standard Weinreb amide chemistries, acid (17) can be converted to methyl ketone (18), which can similarly be converted to a difluoroethyl substituted compound (19) using DAST.

A wide range of substituents can be obtained using Weinreb amide chemistries as shown in Scheme 2b. For example, a Grignard reagent prepared from 3iodoxetane (commercially available from Sigma-Aldrich) can in principle be reacted with the aforementioned Weinreb reagent to form a similar ketone, which can be converted to the difluoro analog (20) using a conventional DAST reagent. This chemical transformation can also be applied to longer chain alkyl difluoro methylene substituents as well as to aryl difluoro methylene substituents.

For some specific heterocycles, an alternative chemical transformation may be used. For example, 2-trimethylthiazole can react with aldehydes, followed by cleavage of the trimethylsilyl (TMS) ether and oxidation of the alcohol to form 2thiazolyl ketone (21) (for an example of this chemical transformation, see Dondoni, A. et al., JOC, 2004, 69, 5023 ), which can be converted to difluoromethylene using the DAST reagent. For the formation of, for example, a trifluoromethyl ketone substituent, the corresponding chemical transformations of silicon can be used, since (trifluoromethyl)trimethylsilane can react with aldehydes to obtain the corresponding carbinol (Cheng, H., et al., Tet. Lett., 2013, 54, 4483). Then, after standard oxidation, trifluoromethyl ketone (22) is obtained.

Examples

General Methods

Spectra ¹ H NMR (nuclear magnetic resonance) are shown at 300 or 400 MHz in solutions in DMSO-d ₆ (δ ^ppm ) using chloroform as a reference standard (7.25 ^ppm ). When peak multiplicities are given, the following abbreviations are used: s (singlet), d (doublet), t (triplet), m (multiplet), bs (broadened singlet), dd (doublet of doublets), dt (doublet of triplets) , q (quartet). Spin-spin coupling constants, if indicated, are given in hertz (Hz).

All reactions were carried out under an inert gas atmosphere of nitrogen or argon, unless otherwise noted (eg, hydrogenation reactions).

Abbreviations:

ACN (from English acetonitrile) - acetonitrile,

DAST (from English diethylaminosulfur trifluoride) - diethylaminosulfur trifluoride, DCM - dichloromethane,

DIPEA (from the English N, N-diisopropylethylamine) -N, N-diisopropylethylamine,

DMAP (from English 4-dimethylaminopyridine)-4-dimethylaminopyridine,

DMSO - dimethyl sulfoxide, h - hours,

NMVS (from the English heteronuclear multiple bond correlation) - heteronuclear correlation spectroscopy of multiple bonds, psi - pounds per square inch,

SO ₃ :pyr - pyridine-sulfur trioxide complex,

TEA (from English triethylamine) - triethylamine,

TEMPO (from English (2,2,6,6-tetramethylpiperidin-1-yl)oxyl)-(2,2,6,6-tetramethylpiperidin-1-yl)oxyl, THF - tetrahydrofuran.

Example 1

Sodium (2S,5R)-7-Oxo-2-(trifluoromethyl)-1,6-diazabicyclo[3.2.1]octan-6-yl sulfate.

- 37 042632

Prepared using essentially the same methodology as given for avibactam (Ball,

M. et al., Organic Process Research and Development, 2016, 20, 1799), except for using (5S)5-(trifluoromethyl)-2-pyrrolidinone as starting compound.

a) tert-Butyl-(5S)-2-oxo-5-(trifluoromethyl)pyrrolidine-1-carboxylate.

A solution of (5S)-5-(trifluoromethyl)-2-pyrrolidinone (commercially available from Manchester Organics) (5 g; 32.7 mmol) in DCM (60 ml) at 0°C was treated with triethylamine (5.5 ml; 4 0 g, 39.2 mmol) and DMAP (0.4 g, 0.1 mmol), then a solution of di-tert-butyl dicarbonate (8.6 g, 39.2 mmol) in DCM (20 ml) for 10 min. After 0.5 h the mixture was partitioned between DCM and 10% aqueous citric acid and the organic extract was washed with water, dried (Na ₂ SO ₄ ) and evaporated to give an oil (9.4 g) which was purified by chromatography on silica with eluting with ethyl acetate (0-25%) in toluene to give an oil (7.9 g; 96%).

M/z 276 (M+Na)+.

b) (3R,6S)-N-(Benzyloxy)-6-(trifluoromethyl)piperidine-3-amine.

For a detailed explanation of the sequence of this reaction, see Ball, M. et al., Organic Process Research and Development, 2016, 20, 1799.

DMSO (10 ml) was added to a mixture of trimethylsulfoxonium iodide (1.6 g; 7.3 mmol) and potassium tert-butoxide (0.72 g; 6.4 mmol) in THF (7 ml). The mixture was stirred at room temperature for 1 hour, then cooled to -12°C (internal temperature). A solution of tert-butyl (5S)-2-oxo-5-(trifluoromethyl)pyrrolidine-1-carboxylate (1.5 g; 5.8 mmol) in THF (4 ml) was added dropwise over 5 min and the mixture was stirred at - 12° C. for 1 hour. The reaction mixture was treated with 20% aqueous ammonium chloride solution (13 ml) and the stirred mixture was allowed to warm to room temperature, then extracted twice with ethyl acetate. The combined extracts were washed with 10% aqueous sodium chloride solution, then the solution was concentrated to approximately 20 ml and used directly in the next step.

(Mass spectroscopy data for this solution corresponded to the formation of (1Z,5S)-5-{[(tert-butoxy)carbonyl]amino}-1-[dimethyl(oxo)-lambda-6-sulfanyliumyl]-6,6,6-trifluorohex -1-en-2-olata.)

The above solution in ethyl acetate (20 ml) was treated with O-benzylhydroxylamine hydrochloride and the mixture was heated at 60°C for 2.75 hours, then left to cool to room temperature, after which it was washed with 10% aqueous sodium chloride solution. This solution was concentrated to approximately 10 ml and used directly in the next step.

(Mass spectroscopy data for this solution corresponded to the formation of tert-butyl-N[(2S,5E/Z)-5-[(benzyloxy)imino]-6-chloro-1,1,1-trifluorohexan-2-yl] carbamate .)

The above solution in ethyl acetate (10 ml) was treated with methanesulfonic acid (1.1 ml; 1.7 g; 17.4 mmol) and the mixture was heated at 45°C for 1 h, then left to cool to room temperature. The mixture was added to a solution of potassium bicarbonate (2.9 g; 29 mmol) in water (10 ml) and stirred at 45°C for 3 hours. After cooling, the phases were separated and the ethyl acetate phase was washed with 10% aqueous sodium chloride solution. The ethyl acetate solution was used as such in the next step.

(Mass spectroscopy data for this solution corresponded to the formation of (3E/Z,6S)-N(benzyloxy)-6-(trifluoromethyl)piperidin-3-imine.)

The above solution in ethyl acetate was cooled to -15°and was added concentrated sulfuric acid (0.6 g; 0.3 ml; 6 mmol). Sodium triacetoxyborohydride (0.5 g; 2.4 mmol) was then added in portions, allowing the temperature to rise from -15° C. to -5° C. over 1 hour. Water was added, followed by concentrated aqueous ammonia (1 ml). The mixture was extracted with ethyl acetate, the organic extract was washed with brine, dried and evaporated to give an oil which was purified by chromatography on silica eluting with ethyl acetate (15-40%) in DCM to give (3R,6S)-N(benzyloxy)-6-(trifluoromethyl )piperidine-3-amine as an oil (184 mg; 12% over 4 steps).

M/z 275 (M+H) ⁺ .

- 38 042632

c) (2S,5R)-6-(Benzyloxy)-2-(trifluoromethyl)-1,6-diazabicyclo[3.2.1]octan-7-one.

Mixture of (3R,6S)-N-(benzyloxy)-6-(trifluoromethyl)piperidine-3-amine (0.18 g; 0.67 mmol) and potassium carbonate (0.53 g; 3.82 mmol) in DCM (20 ml) was treated with triphosgene (0.2 g; 0.67 mmol) at -10°C. After 30 minutes, DMAP (3 mg; 0.03 mmol) was added. The reaction mixture was stirred at room temperature overnight, then washed with water, dried (Na ₂ SO ₄ ) and evaporated. The residue was purified by silica chromatography eluting with an ethyl acetate-DCM gradient to give a colorless solid (0.16 g, 81%).

M/z 301.4 (M+H)+.

d) Sodium (2S,5R)-7-Oxo-2-(trifluoromethyl)-1,6-diazabicyclo[3.2.1]octan-6-yl sulfate.

A solution of (2S,5R)-6-(benzyloxy)-2-(trifluoromethyl)-1,6-diazabicyclo[3.2.1]octan-7-one (163 mg; 0.54 mmol) in isopropanol (5 mL) was treated trimethylamine-sulfur trioxide complex (85 mg; 0.61 mmol), triethylamine (11 mg; 0.11 mmol), 10% palladium on activated carbon (10 mg) and water (0.6 ml). The mixture was hydrogenated under pressure using a balloon for 4.5 hours, then additional trimethylamine-sulfur trioxide complex (82 mg, 0.3 mmol) was added. The mixture was stirred under nitrogen for 2 hours, then filtered and concentrated to a volume of approximately 1.5 ml. The mixture was diluted with water and treated with saturated aqueous sodium bicarbonate (3 ml). The mixture was loaded onto a C18 reverse phase chromatography cartridge (10 g) and eluted with 0-40% ACN in water. Evaporation of product containing fractions gave a white solid which was redissolved in water and repurified by chromatography using the same chromatography conditions. Evaporation of the product containing fractions gave a white solid which was redissolved in water and repurified by chromatography using the same chromatography conditions except that THF was used instead of ACN. Evaporation gave the title compound as a white solid (86 mg; 51%).

¹ H NMR (400 MHz, d6-DMSO) δ 4.09-4.06 (1H, m), 3.89-3.78 (1H, m), 3.24 (1H, d, J=12.0 Hz), 3.13-3.08 (1H, m), 1.92-1.74 (4H, m).

¹⁹ F NMR (376.4 MHz, d ₆ -DMSO) δ 74.7.

LCMS (liquid chromatography combined with mass spectroscopy) (ESI (electrospray ionization) [M-Na] ^- ) 289.1 (97.2%).

Example 2

Sodium (2S,5R)-7-Oxo-2-(difluoromethyl)-1,6-diazabicyclo[3.2.1]octan-6-yl sulfate.

Suspension of ethyl-(2S,5R)-5-[(benzyloxy)amino]piperidine-2-carboxylate in the form of oxalic salt

- 39 042632 acid (15.0 g; 40.7 mmol; purchased from Frapps Chemicals Co., Zhejiang, China, http://www.frappschem.com) in THF (150 ml) at 0°C was treated with a solution of potassium hydrogen carbonate (16.3 g; 163 mmol) in water (150 ml). After 1 h the mixture was extracted with ethyl acetate, the combined extracts were washed with water then brine, dried (Na ₂ SO ₄ ) and evaporated to give ethyl-(2S,5R)-5[(benzyloxy)amino]piperidine-2-carboxylate as a brown oil (10.0 g; 88%).

A solution of ethyl (2S,5R)-5-[(benzyloxy)amino]piperidine-2-carboxylate (5.0 g; 18 mmol) in DCM (100 ml) at 0°C was treated with DIPEA (12.5 ml; 72 mmol), then a solution of triphosgene (2.6 g; 9 mmol) in DCM (10 ml) was added. The mixture was stirred at ambient temperature for 16 h, then saturated aqueous potassium bicarbonate (100 ml) was added. After 1 h, the phases were separated, the DCM phase was washed with water, then with brine, dried (Na ₂ SO ₄ ) and evaporated to give a brown oil (5.0 g; 92%), spectrally corresponding to ethyl-2S,5R)-6- (benzyloxy)-7-oxo-1,6-diazabicyclo[3.2.1]octane-2-carboxylate, and which was used directly without purification.

M/z 305.4 (M+H)+.

b) (2S,5R)-6-(Benzyloxy)-2-(hydroxymethyl)-1,6-diazabicyclo[3.2.1]octan-7-one.

Solution of crude ethyl (2S,5R)-6-(benzyloxy)-7-oxo-1,6-diazabicyclo[3.2.1]octane-2carboxylate (5.0 g; 16.4 mmol) in THF (75 ml) and ethanol (75 ml) was treated at 0°C with a solution of lithium borohydride in THF (4 M; 24 ml; 96 mmol). After 1 hour at room temperature, the mixture was re-cooled to 0°C and another portion of a solution of lithium borohydride in THF (4 M; 12 ml; 48 mmol) was added. After an additional 16 hours at room temperature, the mixture was treated with saturated aqueous potassium monophosphate (200 ml) and the mixture was extracted several times with DCM. The combined DCM extracts were washed with water then brine, dried (Na ₂ SO ₄ ) and evaporated to give a brown oil (5 g). It was purified by chromatography on silica eluting with ethyl acetate (0-100%) in hexane to give a colorless oil (2.2 g; 47% over 2 steps).

M/z 263.3 (M+H) ⁺ .

c) (2S,5R)-6-(Benzyloxy)-7-oxo-1,6-diazabicyclo[3.2.1]octane-2-carbaldehyde.

Solution of (2S,5R)-6-(benzyloxy)-2-(hydroxymethyl)-1,6-diazabicyclo[3.2.1]octan-7-one (1.5 g; 5.7 mmol) in DCM (50 ml ) was treated at 0°C with trichloroisocyanuric acid (1.9 g; 8.6 mmol) and TEMPO (90 mg; 0.6 mmol).

The mixture was stirred at 0°C for 2 h, then filtered through celite, washing with DCM. The combined DCM filtrates were washed with saturated aqueous sodium bicarbonate, brine, dried (Na ₂ SO ₄ ) and evaporated to give an oil (1.5 g; 100%) which was used directly in the next step.

M/z 261.4 (M+H) ⁺ .

d) (2S,5R)-6-(Benzyloxy)-2-(difluoromethyl)-1,6-diazabicyclo[3.2.1]octan-7-one.

A solution of (2S,5R)-6-(benzyloxy)-7-oxo-1,6-diazabicyclo[3.2.1]octane-2-carbaldehyde (1.5 g; 5.7 mmol) in DCM (30 ml) at 0°C was treated with DAST (1.5 ml; 12 mmol). The mixture was stirred at room temperature for 4 h, then the solvent was removed by purging with nitrogen. The residue was dissolved

- 40 042632 in ethyl acetate and added to ice water. The organic phase was separated, washed with saturated aqueous sodium bicarbonate, brine, dried (Na ₂ SO ₄ ) and evaporated to give an oil. This oil was purified by chromatography on silica eluting with 20% ethyl acetate in hexane to give a yellow oil (0.7 g; 44% over 2 steps).

M/z 283.3 (M+H)+.

f) (2S,5R)-2-(Difluoromethyl)-6-hydroxy-1,6-diazabicyclo[3.2.1]octan-7-one.

A solution of (2S,5R)-6-(benzyloxy)-2-(difluoromethyl)-1,6-diazabicyclo[3.2.1]octan-7-one (0.60 g; 2.1 mmol) in methanol (15 ml ) was hydrogenated over 10% palladium on activated carbon (0.60 g) at 100 psi (689 kPa) in a steel canister. After 4 hours, the mixture was filtered through celite, washing with methanol, and the combined filtrates were evaporated to give a white solid (0.4 g; 100%), which was used directly in the next step.

M/z 193.3 (M+H) ⁺ .

f) Tetrabutylammonium (2S,5R)-2-(Difluoromethyl)-7-oxo-1,6-diazabicyclo[3.2.1]octan-6-ylsulfate.

A solution of (2S,5R)-2-(difluoromethyl)-6-hydroxy-1,6-diazabicyclo[3.2.1]octan-7-one (0.40 g; 2.1 mmol) in DCM (30 ml) at 0°C was treated with TEA (1.1 ml; 8.3 mmol), then the pyridine sulfur trioxide complex (0.67 g; 4.2 mmol) was added. After 4 hours at room temperature, a solution of ammonium n-tetrabutyl acetate (0.94 g; 3.1 mmol) in water (20 ml) was added. After 2 hours, additional DCM was added and phase separation was performed. The DCM phase was washed with water, dried (Na ₂ SO ₄ ) and evaporated. The residue was purified by chromatography on silica eluting with ethyl acetate (0-100%) in hexane then 4% methanol in DCM to give a colorless oil (0.45 g; 42% over 2 steps).

M/z 271.4 (M) ^- .

g) Sodium (2S,5R)-7-Oxo-2-(difluoromethyl)-1,6-diazabicyclo[3.2.1]octan-6-yl sulfate.

A solution of tetrabutylammonium (2S,5R)-2-(difluoromethyl)-7-oxo-1,6-diazabicyclo[3.2.1]octan-6-yl sulfate (0.45 g; 0.88 mmol) in water (10 ml) treated with Dowex™ Na resin (1 g). After 1 hour, the mixture was filtered through a pad of Dowex™ Na resin. The combined filtrates were passed through a second pack of Dowex™ Na resin, washing with water (5 ml), then the combined filtrates were freeze-dried to give the title compound as a white solid (202 mg; 78%).

¹ H NMR (400 MHz, d ₆ -DMSO) δ 6.24 (1H, t, J=4.4 Hz, CHF2), 4.06 (1H, m), 3.40 (1H, m), 3.20 (1H, m), 3.05 (1Н, m), 1.95-1.75 (4Н, m).

LCMS (ESI [M-Na] ^- ) 271.1.

Example 3

Sodium (2S,5R)-2-(Dichloromethyl)-7-oxo-1,6-diazabicyclo[3.2.1]octan-6-yl sulfate.

a) (2S,5R)-6-(Benzyloxy)-2-(dichloromethyl)-1,6-diazabicyclo[3.2.1]octan-7-one.

- 41 042632

A solution of (2S,5R)-6-(benzyloxy)-7-oxo-1,6-diazabicyclo[3.2.1]octane-2-carbaldehyde (2.2 g; 8.4 mmol) in DCM (100 mL) was treated phosphorus pentachloride (3.5 g; 16.9 mmol). After 16 h the mixture was diluted with DCM, washed with ice water, brine, dried (Na ₂ SO ₄ ) and evaporated. The residue was purified by chromatography on silica eluting with ethyl acetate (0-20%) to give 3 separate fractions. Fractions 1 and 3 did not contain the desired compound, and fraction 2 (TLC (Thin Layer Chromatography): R _f (retention factor) 0.4 in 20% ethyl acetate/hexane) was a white solid (130 mg; 5%), spectral analysis data corresponding to the desired compound. In particular, studies using HMBS NMR showed a close spatial proximity between the carbonyl carbon and the C2-H proton, thereby defining the -CC12H substituent as axial and the C2 chirality as S, as in the diagram.

M/z 315.4; 317.3 (M+H)+.

b) (2S,5R)-2-(Dichloromethyl)-6-hydroxy-1,6-diazabicyclo[3.2.1]octan-7-one.

A solution of 6-(benzyloxy)-2-(dichloromethyl)-1,6-diazabicyclo[3.2.1]octan-7-one (130 mg; 0.4 mmol) in methanol (10 ml) was treated with 10% palladium on activated charcoal (130 mg) and hydrogenated (using a pressurized balloon) for 2 hours. The mixture was filtered through celite, washing with methanol. The combined filtrates were evaporated to give an off-white solid (85 mg; 92%) which was used directly in the next step.

M/z 225.4; 227.3 (M+H) ⁺ .

c) Tetrabutylammonium (2S,5R)-2-(Dichloromethyl)-7-oxo-1,6-diazabicyclo[3.2.1]octan-6-ylsulfate.

A solution of 2-(dichloromethyl)-6-hydroxy-1,6-diazabicyclo[3.2.1]octan-7-one (85 mg; 0.37 mmol) in DCM (10 ml) was treated with TEA (0.2 ml; 0 .8 mmol) and pyridine-sulfur trioxide complex (0.12 g; 0.76 mmol) at 0°C. The mixture was stirred at room temperature for 4 hours, then evaporated to dryness. The residue was redissolved in dimethylformamide (DMF) (2 ml), treated with TEA (0.4 ml; 1.6 mmol) and additional pyridine-sulfur trioxide complex (0.12 g; 0.76 mmol). The mixture was stirred at room temperature for 2 h, then a solution of ammonium n-tetrabutyl acetate (0.28 g; 0.8 mmol) in water (10 ml) was added. After 2 h the mixture was diluted with DCM, the organic extract was washed with water then brine, dried (Na ₂ SO ₄ ), then evaporated to give an oil. This oil was purified by chromatography on silica eluting with ethyl acetate (0-100%) in hexane followed by 6% methanol in DCM to give a colorless oil (68 mg; 33%).

M/z 304.4; 306.3 (M) ^- .

d) sodium (2S,5R)-2-(dichloromethyl)-7-oxo-1,6-diazabicyclo[3.2.1]octan-6-yl sulfate.

A solution of (2S,5R)-2-(dichloromethyl)-7-oxo-1,6-diazabicyclo[3.2.1]octan-6-ylsulfate tetrabutylammonium (68 mg; 0.12 mmol) in water (2 ml) and ACN (2 ml) was treated with Dowex™ Na resin (0.5 g). After 1 hour, the mixture was filtered through a pad of Dowex™ Na resin. The combined filtrates were passed through a second pack of Dowex™ Na resin, washing with water-ACN (1 ml:1 ml). The filtrates were passed through a second pack of Dowex™ Na resin, then the combined filtrates were freeze-dried to give the title compound as a white solid (36 mg; 88%).

¹ H NMR (400 MHz, d ₆ -DMSO) δ 6.25 (1H, d, J=0.8 Hz, CHCl ₂ ), 4.02 (1H, m), 3.45 (1H, m), 3.22 (1H, d) , 2.95 (1Н, m), 1.95–1.75 (4Н, m).

LCMS (ESI [M-Na]-) 303.4; 305.4.

Example 4

Sodium (2S,5R)-2-(Fluoromethyl)-7-oxo-1,6-diazabicyclo[3.2.1]octan-6-yl sulfate.

Example 4 was prepared from the key alcohol intermediate via fluoromethyl substituent conversion using standard DAST workup (see, e.g., Collet, C. et al., Bioorg. Med. Chem. Lett, 2017, 25, 5603).

The remainder of this synthesis (during debenzylation, sulfonation and sodium salt formation) followed the procedures outlined above.

M/z=253.0 (M-Na)'.

¹ H NMR (500 MHz, d ₆ -DMSO) δ 4.71-4.48 (2H, m, CH ₂ F ₂ ), 3.98 (1H, m), 3.42 (1H, m), 3.19 (1H, m), 2.92 ( 1Н, m), 1.84 (1Н, m), 1.78-1.66 (2Н, m), 1.51 (1Н, m).

Example 5

Sodium (2S,5R)-2-(1,1-difluoroethyl)-7-oxo-1,6-diazabicyclo[3.2.1]octan-6-yl sulfate.

Example 5 was prepared by converting the acid to Weinreb amide, forming methyl ketone using methylmagnesium bromide, then converting methyl ketone by introducing a fluoroethyl substituent using DAST treatment (Wityak, J., et al., J. Medicinal Chemistry, 2015, 58, 2967) .

The remainder of this synthesis (during debenzylation, sulfonation and sodium salt formation) followed the procedures outlined above.

M/z=285.0 (M-Na)-.

¹ H NMR (500 MHz, d ₆ -DMSO) δ 4.02 (1H, m), 3.40 (1H, m), 3.18 (1H, m), 3.02 (1H, m), 1.89-1.79 (2H, m), 1.78-1.65 (2H, m), 1.72 (3H, m), 1.51 (1H, m).

Example 6

Sodium (2S,5R)-2-(Chloromethyl)-7-oxo-1,6-diazabicyclo[3.2.1]octan-6-yl sulfate.

The compound of Example 6 was prepared by reacting the alcohol with methanesulfonyl chloride in the presence of triethylamine to form the corresponding mesylate, then substitution using ammonium tetra-n-butyl chloride to give the chloromethyl-containing intermediate.

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The remainder of this synthesis (during debenzylation, sulfonation and sodium salt formation) followed the procedures outlined above.

M/z=268.8 (M-Na) ^- .

1Н NMR (500 MHz, de-DMSO) δ 3.99 (1Н, m), 3.88 (1Н, m), 3.81 (1Н, m), 3.32 (1Н, m), 3.19 (1Н, m), 2.89 (1Н, m), 1.83-1.69 (3Н, m), 1.52 (1Н, m).

Example 7

Sodium (2R,5R)-7-Oxo-2-[(trifluoromethyl)sulfanyl]-1,6-diazabicyclo[3.2.1]octan-6-yl sulfate.

The compound of Example 7 was prepared using a decarboxylation approach using a similar methodology to the introduction of halogens, but using silver(I) trifluoromethanethiolate, following the conditions described by Liu C. et al. (RSC Advances, 2017, 7, 880).

The remainder of this synthesis (during debenzylation, sulfonation, and sodium salt formation) followed the procedures outlined above, with the difference that the benzyl group was removed using titanium tetrachloride, since the hydrogenation was unsuccessful.

M/z=320.9 (M-Na) ^- .

1H NMR (500 MHz, de-DMSO) δ 5.04 (1H, dd, J=12.5 Hz; J=4.5 Hz; CHSCF3), 4.01 (1H, d, J=3.0 Hz), 3.23 ( 2Н, m), 2.01 (2Н, m), 1.89–1.72 (2Н, m).

Example 8

Sodium (2S,5R)-7-Oxo-2-[(trifluoromethoxy)methyl]-1,6-diazabicyclo[3.2.1]octan-6-yl sulfate.

Example 8 was prepared by trifluoromethylation of an alcohol using trifluoromethyltrimethylsilane (TMSCF3) using silver(1) triflate and Selectfluor according to the conditions of Liu J.-B. and others (Organic Letters, 2017, 17, 5048).

The remainder of this synthesis (during debenzylation, sulfonation and sodium salt formation) followed the procedures outlined above.

M/z=318.9 (M-Na) ^- .

1H NMR (500 MHz, de-DMSO) δ 4.32 (1H, dd, J=10.5 Hz; J=9.5 Hz), 4.16 (1H, dd, J=10.5 Hz; J=5.5 Hz), 3.98 (1Н, d, J=3 Hz), 3.45 (1Н, m), 3.21 (1Н, m), 2.91 (1Н, m), 1.85-1.69 (3Н, m), 1.48 (1Н, m ).

- 44 042632

Example 9

(2S,5R)-2-[Difluoro(1,3-thiazol-2-yl)methyl]-7-oxo-1,6-diazabicyclo[3.2.1]octan-6-ylsulfate sodium.

Example 9 was prepared from an aldehyde which was converted to a thiazolyl ketone following the methodology described by Dondoni A. et al. (2004, Journal of Organic Chemistry, 69, 5023), namely by in situ formation of the thiazole anion and reaction with this aldehyde; capture with acetic anhydride to obtain acetate; hydrolysis of the acetate followed by a Svern-like oxidation to form a ketone. This ketone was reacted with DAST in the usual manner to form geminal difluoride.

The remainder of this synthesis (during debenzylation, sulfonation, and sodium salt formation) followed the procedures outlined above, with the difference that the benzyl group was removed using titanium tetrachloride, since the hydrogenation was unsuccessful.

M/z=353.9 (M-Na) ^- .

¹ H NMR (500 MHz, d6-DMSO) δ 8.04 (2H, s), 4.05-3.97 (2H, m), 3.24 (1H, m), 3.01 (1H, m), 2.03-1.98 (1H, m) , 1.88-1.77 (3H, m).

Example 10

Sodium (2R,5R)-2-Fluoro-7-oxo-1,6-diazabicyclo[3.2.1]octan-6-yl sulfate.

a) (2S,5R)-6-(Benzyloxy)-7-oxo-1,6-diazabicyclo[3.2.1]octane-2-carboxylic acid.

To a solution of commercially available ethyl (2S,5R)-6-(benzyloxy)-7-oxo-1,6-diazabicyclo[3.2.1]octane2-carboxylate (7.0 g; 23.0 mmol) in acetone :water (1:1, 120 mL) LiOH-H2O (0.97 g; 23.0 mmol) was added at 0°C. The resulting reaction mixture was stirred at room temperature for 2 h, then the reaction mixture was diluted with water (50 ml) and washed with EtOAc (2x100 ml). The aqueous layer was acidified with 1N. HCl solution (to pH ~3) and then extracted with EtOAc (2x100 ml). The combined organic phases were washed with water, brine, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated to give (2S,5R)-6-(benzyloxy)-7-oxo-1,6-diazabicyclo[3.2.1]octane-2 α-carboxylic acid (5 g; 79%) as a white solid, which was used in the next step without purification.

M/z=277.1 (M+H)+.

b) (2R,5R)-6-(Benzyloxy)-2-fluoro-1,6-diazabicyclo[3.2.1]octan-7-one (A) and (2S,5R)-6-(benzyloxy)2- fluoro-1,6-diazabicyclo[3.2.1]octan-7-one (B).

- 45 042632

To a stirred solution of (2S,5R)-6-(benzyloxy)-7-oxo-1,6-diazabicyclo[3.2.1]octane-2-

carboxylic acid (2.5 g; 9.04 mmol) in acetone:water (4:1, 100 ml) were added Selectfluor™ (6.4 g; 18.0 mmol) and AgNO ₃ (153 mg; 0.90 mmol) at room temperature. The resulting reaction mixture was heated at 50°C for 3 h, then evaporated. The resulting residue was treated with EtOAc (100 ml), filtered through a Celite pad and the pad washed with EtOAc (10 ml). The filtrate was washed with NaHCO ₃ solution (50 ml), water (50 ml) and brine (50 ml). The organic layer was dried over anhydrous Na ₂ SO ₄ , filtered and concentrated to give an oil. This oil was purified by silica gel chromatography using 10% EtOAc in hexane as eluent to give (2R,5R)-6-(benzyloxy)-2-fluoro1,6-diazabicyclo[3.2.1]octan-7-one ( A) (550 mg; 24%) as a pale yellow viscous oil. Further elution with 50-60% EtOAc in hexane gave (2S,5R)-6-(benzyloxy)-2-fluoro-1,6diazabicyclo[3.2.1]octan-7-one (B) (300 mg; 13%) as a pale yellow solid.

(2R,5R)-6-(Benzyloxy)-2-fluoro-1,6-diazabicyclo[3.2.1]octan-7-one (A), m/z=251.1 (M+H)+.

Experiments using NMR showed that the values of the spin-spin interaction constants for the H atom at the same carbon atom to which the F atom is connected were 45.0 Hz (for interaction with F) and 4.5 Hz (for interaction with axial proton on the adjacent carbon atom), thereby confirming that this H atom is located equatorially and the molecule has the shown stereochemical configuration.

(2S,5R)-6-(Benzyloxy)-2-fluoro-1,6-diazabicyclo[3.2.1]octan-7-one (B), m/z=251.1 (M+H)+.

Experiments using NMR showed that the values of the spin-spin interaction constants for the H atom at the same carbon atom to which the F atom is connected were 44.0 Hz (for interaction with F) and 10.5 Hz (for interaction with axial proton on the adjacent carbon atom), thereby confirming that this H atom is located axially, and the molecule has the shown stereochemical configuration.

c) (2R,5R)-2-Fluoro-6-hydroxy-1,6-diazabicyclo[3.2.1]octan-7-one.

10 % Pd/C (300 mg). The reaction mixture was hydrogenated at room temperature for 1 hour using a pressurized hydrogen balloon. The reaction mixture was filtered through a pad of celite, washing with methanol (10 ml). The filtrate was evaporated to give (2R,5R)-2-fluoro6-hydroxy-1,6-diazabicyclo[3.2.1]octan-7-one (180 mg; 94%) as an off-white solid which was used in the next step without cleaning.

M/z=161.0 (M+H) ⁺ .

d) tetrabutylammonium (2R,5R)-2-fluoro-7-oxo-1,6-diazabicyclo[3.2.1]octan-6-yl sulfate.

TEA (1 .5 ml; 11.2 mmol), then SO ₃ :Py complex (1.07 g; 6.74 mmol) was added at 0° C. and stirred at room temperature for 4 h. 7 g, 8.99 mmol) in water (20 ml) and stirred at room temperature for 2 hours. The reaction mixture was diluted with DCM (50 ml) and the organic layer was separated. The organic layer was washed with water (5x25 ml), dried over Na ₂ SO ₄ , filtered and evaporated. The resulting residue was purified by chromatography on silica eluting with ethyl acetate (0-100%) in hexane followed by 5% methanol in DCM to give (2R,5R)-2-fluoro-7-oxo-1,6-diazabicyclo[3.2 .1] tetrabutylammonium octan-6yl sulfate (220 mg; 40% in 2 steps) as a colorless oil.

M/z=239.0 (M-NBU4)·.

(a) Sodium (2R,5R)-2-Fluoro-7-oxo-1,6-diazabicyclo[3.2.1]octan-6-yl sulfate.

A stirred solution of rabutylammonium (2R,5C)-2-fluoro-7-oxo-1,6-diazabicyclo[3.2.1]octan-6-yl sulfate (220 mg; 0.45 mmol) in water (10 ml ) were treated with Dowex™ Na resin (1 g). After 1 hour, the mixture was filtered through a pad of Dowex™ Na resin, rinsing with H 2 O (5 ml). The combined filtrate was re-treated with Dowex™ Na resin (1 g) for 1 hour, filtered through a resin pack.

Dowex™ Na, washing with H2O (5 ml). This process was repeated 3 more times. The combined filtrates were freeze-dried to give the title compound (90 mg; 75%) as an off-white solid.

M/z=239.0 (M-Na) ^- .

¹ H NMR (500 MHz, d6-DMSO) δ 5.35 (1H, ddd, J=46 Hz; 5 Hz; 2.5 Hz, CHF), 4.08-4.06 (1H, m), 3.263.24 (1H, m ), 3.06–3.04 (1Н, m), 1.99–1.68 (4Н, m).

Example 11.

Sodium (2S,5R)-2-Fluoro-7-oxo-1,6-diazabicyclo[3.2.1]octan-6-yl sulfate.

NJ _Λ I О //---SO ^Na

Ο 0'7 o

a) (2S,5R)-2-Fluoro-6-hydroxy-1,6-diazabicyclo[3.2.1]octan-7-one.

J--N

Oh Oh

To a solution of (2S,5R)-6-(benzyloxy)-2-fluoro-1,6-diazabicyclo[3.2.1]octan-7-one (250 mg; 0.99 mmol) (see compound (B) from example 10) in methanol (10 ml) was added 10% Pd/C (250 mg). The reaction mixture was hydrogenated at room temperature for 1 hour using a pressurized hydrogen balloon. The reaction mixture was filtered through a pad of celite and the pad was washed with methanol (5 ml). The filtrate was evaporated to give (2S,5R)-2-fluoro-6-hydroxy-1,6-diazabicyclo[3.2.1]octan-7-one (130 mg; 81%) as an off-white solid which was used in the next stages without purification.

M/z=161.1 (M+H)+.

b) Tetrabutylammonium (2S,5R)-2-Fluoro-7-oxo-1,6-diazabicyclo[3.2.1]octan-6-yl sulfate.

TEA (1 14 ml, 8.11 mmol), then SO ₃ :Py complex (775 mg, 4.87 mmol) at 0°C. The mixture was stirred at room temperature for 4 hours. A solution of ammonium n-tetrabutyl acetate (1.9 g; 6.49 mmol) in water (30 ml) was added and the mixture was stirred at room temperature for 2 hours. The reaction mixture was diluted with DCM (50 ml) and the organic layer was separated. The DCM layer was washed with water (5x25 ml), dried over Na ₂ SO ₄ , filtered and evaporated. The resulting residue was purified by chromatography on silica eluting with 0-100% EtOAc in hexane followed by 5% methanol in DCM to give (2S,5R)-2-fluoro-7-oxo-1,6-diazabicyclo[3.2.1 tetrabutylammonium octan-6-yl sulfate (60 mg; 15% in 2 steps) as a colorless oil.

M/z=239.1 (M-NBU4) ^- .

c) sodium (2S,5R)-2-fluoro-7-oxo-1,6-diazabicyclo[3.2.1]octan-6-yl sulfate.

A stirred solution of tetrabutylammonium (2R,5R)-2-fluoro-7-oxo-1,6-diazabicyclo[3.2.1]octan-6-ylsulfate (60 mg; 0.12 mmol) in CH ₃ CN:H ₂ O (1:1, 4 ml) was treated with Dowex™ Na resin (1 g). After 1 h, the mixture was filtered through a pad of Dowex™ Na resin and washed with CH ₃ CN:H ₂ O (1:1, 2 ml). The combined filtrate was re-treated with Dowex™ Na resin (1 g) for 30 min, filtered through a pad of Dowex™ Na resin and washed with CH ₃ CN:H ₂ O (1:1, 2 ml). This process was repeated 3 more times. The combined filtrates were freeze-dried to give the title compound as an off-white solid.

M/z=238.9 (M-Na) ^- .

¹ H NMR (500 MHz, d6-DMSO) δ 5.27 (1H, ddd, J=44 Hz; J=10.5 Hz; J=4.5 Hz, CHF), 3.96-3.94 (1H, m), 3.24 -3.21 (1Н, m), 3.01 (1Н, J=12 Hz), 2.06-2.01 (2Н, m), 1.73-1.67 (1Н, m), 1.61-1.57 (1Н, m).

Example 12.

Sodium (2R,5R)-2-Chloro-7-oxo-1,6-diazabicyclo[3.2.1]octan-6-yl sulfate.

- 47 042632

a) (2R,5R)-6-(Benzyloxy)-2-chloro-1,6-diazabicyclo[3.2.1]octan-7-one.

^S 'P

N. L

To a solution of (2S,5R)-6-(benzyloxy)-7-oxo-1,6-diazabicyclo[3.2.1]octane-2-carboxylic acid (1 g; 3.62 mmol) in DMF (20 ml) and AcOH (4 ml) was added N-chlorosuccinimide (4.83 g; 36.2 mmol) at room temperature. The reaction mixture was purged with nitrogen gas for 5 minutes. Pb(OAc) ₄ (2.4 g; 5.43 mmol) was then added and the reaction mixture was purged with nitrogen gas for 5 minutes. The reaction mixture was heated at 60°C for 4 h and then treated with a saturated solution of K ₂ CO ₃ at room temperature. The mixture was extracted with diethyl ether (2x50 ml), the combined organic layer was washed with brine (20 ml), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated to give the crude product. This product was purified by chromatography on silica using 10% EtOAc in hexane as eluent to give (2R,5R)-6-(benzyloxy)-2-chloro1,6-diazabicyclo[3.2.1]octan-7-one (270 mg; 28%) as a light yellow oil.

M/z=267.0 (M+H)+.

Experiments using NMR showed that the value of the spin-spin coupling constant for the H atom at the same carbon atom to which the Cl atom is connected was 5.5 Hz (for interaction with the axial proton on the neighboring carbon atom), thereby confirming that this H atom is located equatorially and the molecule has the stereochemical configuration shown.

b) (2R,5R)-2-Chloro-6-hydroxy-1,6-diazabicyclo[3.2.1]octan-7-one.

J--N

OH OH

10%- ny Pd/C (220 mg). The reaction mixture was hydrogenated at room temperature for 1 hour using a pressurized hydrogen balloon. The reaction mixture was filtered through a pad of celite and the pad was washed with methanol (10 ml). The filtrate was evaporated to give (2R,5R)-2-chloro-6-hydroxy-1,6-diazabicyclo[3.2.1]octan-7-one (150 mg) as an off-white solid which was used without purification.

M/z=177.0 (M+H)+.

c) Tetrabutylammonium (2R,5R)-2-Chloro-7-oxo-1,6-diazabicyclo[3.2.1]octan-6-yl sulfate.

To a stirred solution of (2R,5R)-2-chloro-6-hydroxy-1,6-diazabicyclo[3.2.1]octan-7-one (150 mg; 0.849 mmol) in DCM (20 ml) 8 ml, 12.7 mmol), then SO ₃ :Py complex (1.35 g, 8.49 mmol) at 0° C. and stirred at room temperature for 3 h. ammonium acetate (2.56 g; 8.49 mmol) in water (20 ml) and stirred at room temperature for 3 hours. The reaction mixture was diluted with DCM (50 ml) and the phases were separated. The organic extract was washed with water (5x25 ml), dried over Na ₂ SO ₄ , filtered and evaporated. The residue was purified by chromatography on silica eluting with ethyl acetate (0-100%) in hexane followed by 5% methanol in DCM to give (2R,5R)-2-chloro-7-oxo-1,6-diazabicyclo[3.2. 1]Tetrabutylammonium octan-6-yl sulfate (130 mg; 30% over 2 steps) as a colorless oil.

M/z=254.9 (M-NBu4).

d) Sodium (2R,5R)-2-Chloro-7-oxo-1,6-diazabicyclo[3.2.1]octan-6-yl sulfate.

A solution of (2R,5R)-2-chloro-7-oxo-1,6-diazabicyclo[3.2.1]octan-6-ylsulphate tetrabutylammonium (130 mg; 0.26 mmol) in CH ₃ CN:H ₂ O ( 1:1, 3 ml) were treated with Dowex™ Na resin (1 g). After 30 minutes, the mixture was filtered through a pad of Dowex™ Na resin and washed with CH ₃ CN:H ₂ O (1:1, 1 ml). Merged

- 48 042632 The filtrate was re-treated with Dowex™ Na resin (1 g) for 30 min, filtered through a pad of Dowex™ Na resin and washed with CH ₃ CN:H ₂ O (1:1, 1 ml). This process was repeated 3 more times. The combined filtrates were freeze dried and the resulting compound was purified by preparative high performance liquid chromatography (HPLC) [X-SELECT-C18 (150x19), 5 μm; mobile phase: H2O:MeCN]. The collected fractions were freeze-dried to give the title compound (5.0 mg, 7%) as an off-white solid.

M/z=255.0 (M-Na)-.

¹ H NMR (500 MHz, d6-DMSO) δ 5.51 (1H, dd, J=6.0 Hz; J=2.0 Hz, CHCl), 4.10 (1H, J=3.0 Hz), 3.51-3.49 (1Н, m), 3.07-3.05 (1Н, m), 2.26-2.18 (1Н, m), 1.94-1.80 (3Н, m).

Biological activity

Experiments were carried out to determine (1) the inhibitory activity of the compounds of the invention against serine β-lactamase enzymes from different classes;

(2) enhancing the action of β-lactam antibiotics with these compounds against strains expressing SBL enzymes.

Details of the protocols used for each set of experiments are provided below.

Enzyme inhibition

In Vitro Enzyme Inhibition Assays

Enzyme inhibition assays were performed using solutions of purified SBL enzymes from Enterobacter cloacae (such as TEM-1; AmpC; CTX-M15; KPC-2; OXA-48) in 10 mM HEPES buffer (4-(2-hydroxyethyl)- 1-piperazineethanesulfonic acid), pH 7.5, in 96-well microtiter plates. Nitrocephin was used as a substrate (100 μM for TEM-1, AmpC, KPC-2, OXA-48; and 50 μM for CTX-M15). Its hydrolysis progress was monitored at 482 nm every 30 s for 12 min after an initial incubation period of 10 min at 30° C. using a Perkin Elmer Envision UV fluorescence plate reader. Data on the rate of hydrolysis in the presence of a number of inhibitors were analyzed and IC ₅₀ values (concentration causing 50% inhibition) were determined for each compound using software from the Dotmatics database.

Compounds were diluted in DMSO.

Shown below are the average IC ₅₀ results for inhibition of a selected panel of SBL enzymes. The data is grouped as follows.

TEM-1: IC ₅₀ less than 3 nM (A); 3-10 nM (B); more than 10 nM (C).

KPC-2: IC ₅₀ less than 10 nM (A); more than 10 nM (B).

AmpC: IC ₅₀ less than 10 nM (A); 10-40 nM (B); 40-60 nM (C); greater than 60 nM (D).

OXA-48: IC ₅₀ less than 20 nM (A); 20-100 nM (B); 100-200 nM (C); 200-300 nM (D); more than 300 nM (E).

CTX-M15: IC ₅₀ less than 5 nM (A); 5-20 nM (B); 20-100 nM (C); 100-200 nM (D).

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Compound Deputy R KPC-2 Amp-C OXA-48 CTX-M15 Avibactam conh ₂ 7.8(A) 8.0(A) 296.6(D) 1.4(A) Example 1 _CF3 (A) (A) (A) (B) Example 2 chf ₂ (A) (B) (B) (B) Example 3 CHCI ₂ (A) (C) (C) (C) Example 4 CH ₂ F (B) (A) (D) (B) Example 5 cf ₂ ch ₃ (B) (C) (B) (B) Example 6 _CH2CI (B) (B) (D) N/D Example 7 CF _3S (B) (C) (D) N/D Example 8 CH ₂ OCF ₃ (A) (C) (D) (D) Example 9 СР ₂ -2-thiazolyl (A) (A) (D) (C) Example 10 F(R) (B) (B) (D) N/D Example 11 F(S) (B) (D) (E) N/D Example 12 Cl (B) (A) (E) N/D

N / D (from English not determined) means not determined.

Compound Deputy R TEM-1 Avibactam CONH ₂ 4.7 (V) Example 10 F(R) (A) Example 11 F(S) (WITH) Example 12 Cl (A)

Antimicrobial susceptibility testing Antibiotic activity of β-lactam antibiotics against SBL/ESBL-expressing bacteria in the presence of compounds of the invention

Experiments were performed using the broth microdilution method according to protocols M07-A8 approved by the Clinical and Laboratory Standards Institute (CLSI). Serial dilutions of the β-lactam antibiotic meropenem were prepared in 96-well plates using cation-adjusted Mueller-Hinton broth (CAMHB); the concentration range was set from 0.03 mg/l to 512 mg/l. The turbidity of the bacterial inoculum of each strain (clinical isolates) was corrected to a turbidity standard of 0.5 McFarland in physiological serum (0.9% NaCl), then diluted 1:100 in CAMHB and added to each well to give a final number of bacterial cells of 5 x 10 ⁵ cfu /well. After incubation for 18-20 hours in a heated chamber at 37° C., growth inhibition was assessed based on the absence of any bacterial growth.

The minimum inhibitory concentration (MIC) is taken to be the lowest concentration of antibiotic at which the tested microorganism does not show visible growth; the results were confirmed by measuring the optical density (OD) at 600 nm on a spectrophotometer.

Compounds of the invention were tested at a constant concentration of 4 μg/ml. The clinical strains for these meropenem (mero) enhancement experiments were NTBC091.1 (E. coli strain expressing KPC-2, TEM-1); NTBC093 (E. cloacae strain expressing KPC-2, TEM-1); NTBC096.1 (K. pneumoniae strain expressing OXA-181 and SHV-11); NTBC099 (K. pneumoniae strain expressing KPC-3, SHV-11 and TEM-1); NTBC189 (K. pneumoniae strain expressing TEM-OSBL(b), CTX-M-14, OXA-48(c)).

- 50 042632

Strain number Enzyme(s) p-lactamase(s) Classification MIC for mero, µg/mL, no inhibitor MIC for mero + conn. example 1, 4 µg/ml MIC for mero + conn. example 2, 4 µg/ml NTBC091.1 KRS-2 + TEM-1 E. coli 4 0.03 0.03 NTBO093 KRS-2 + TEM-1 E. cloacae 8 0.06 0.06 NTBC096.1 OXA-181 + SHV-11 TO. pneumoniae 32 4 4 NTBC099 KRS-3 + SHV- 11 + TEM-1 TO. pneumoniae 256 2 1 NTBC189 TEM-OSBL(b) + CTX-M + OXA- 48(c) TO. pneumoniae 32 2 2

Data for other compounds of the invention are shown in the following table. In this table, the data are grouped as follows (data for the compounds of examples 1 and 2 are given for ease of comparison): MIC values less than 1 μg/ml are indicated (A); MIC values of 1 or 2 µg/mL are indicated (B); MIC values of 4 μg/mL are indicated (C); MIC values 8 µg/mL (D); MIC values greater than 8 μg/ml are indicated (E).

Number Enzyme(s) p-lactamase(s) MIC to MIC to MIC d schya M1Sdl I MIC for MIC strain mero + mero mero mero MIC for avibactam, + comp. + connection + connection mero, 4 µg/ml example 1, example 2, example 3, µg/ml, 4 µg/ml 4 µg/ml 4 µg/ml without inhibitor mero for + connection mero example 4, + Comm. 4 µg/ml example a 5.4 µg/ml NTBC091.1 KRS-2 + TEM-1 4 0.06 (A) (. A) (A) (A) (A) (A) NTBC093 KRS-2 + TEM-1 8 0.03 (А) С A) (A) (A) (A) (A) NTBC096.1 OXA-181 + SHV-11 32 1 (B) (C) (C) (E) (IN) (E) NTBC099 NTBC189 KRS-3 + SHV-11 + TEM-1 256 0.25(A) (B) (B) (B) TEM-OSBL(b) + CTX-M + OXA- 32 0.25 (A) (B) (B) N/D 48(c) (IN) N/D (IN) N/D Strain number Enzyme(s) p-lactamase(s) MIC for mero + conn. example 6, 4 µg/ml MIC for mero + conn. example 7, 4 µg/ml MIC for mero + conn. example 8, 4 µg/ml MIC for mero + conn. example 9, 4 µg/ml MIC for mero + conn. example 10, 4 µg/ml MIC for mero + conn. example 11, 4 µg/ml MIC for mero + conn. example 12, 4 µg/ml NTBC091.1 KRS-2 + TEM-1 (A) (A) (A) (A) (A) (A) (A) NTBC093 KRS-2 + TEM-1 (A) (IN) (A) (A) (A) (IN) (A) NTBC096.1 OXA-181 + SHV-11 (E) (E) (E) (E) (A) (D) (D) NTBC099 KRS-3 + SHV-11 +TEM-1 (IN) (E) (E) (D) (IN) (E) (E) NTBC189 TEM-OSBL(b) + CTX-M + OXA-48(c) N/D N/D N/D N/D (A) (WITH) (WITH)

The compounds of the invention are highly active. For example, the combination of meropenem and the compound of example 1 has a lower MIC than the combination of meropenem and avibactam for strain NTBC091.1. The combination of meropenem and the compound of example 2 has a lower MIC than the combination of meropenem and avibactam with respect to strain NTBC091.1. The combination of meropenem and the compound of Example 10 has a lower MIC than the combination of meropenem and avibactam for strains NTBC091.1, NTBC096.1 and NTBC189, and a MIC similar to that of the combination of meropenem and avibactam for strain NTBC093. The combination of meropenem and the compound of example 12 has a lower MIC than the combination of meropenem and avibactam for strain NTBC091.1.

- 51 042632

Additional data on MIC

Further, the compound of example 10 was tested against a large panel of clinical strains of enterobacteria resistant to beta-lactam antibiotics such as meropenem, cefepime and ceftazidime because they produce OXA or KPC variants of the SBL enzymes.

In FIG. 1 shows data showing the effect of the compound of Example 10 on two panels of clinical strains of OXA-positive enterobacteria resistant to beta-lactam antibiotics such as meropenem, cefepime, and ceftazidime. The MIC data is shown in Fig. 1 in standard cumulative form.

In FIG. 1A shows that, for example, at an MIC of 1 μg/ml, 0% of the strains are susceptible to meropenem as monotherapy (MEM). However, the proportion of strains susceptible to various antibiotics at a concentration of 1 μg/ml increases to about 33% (in the case of a clinical combination of the VNRX-5133 inhibitor and the antibiotic cefepime, CEF/VNRX); 63% (in case of clinical combination of avibactam inhibitor and antibiotic ceftazidime, CAZ/AVI); over 90% (in the case of a combination of the inhibitor from example 10 and the antibiotic meropenem).

Similarly, the data of FIG. 1B, which refers to a more extended panel of clinical strains of OXA-positive enterobacteria resistant to beta-lactam antibiotics such as meropenem, cefepime, and ceftazidime, confirm that meropenem exhibits weak activity (MIC ₅₀ is 32 µg/mL; MIC90 is greater than 32 µg /ml). Addition of the compound of example 10 at a concentration of 4 μg/ml restores susceptibility to meropenem (MIC ₅₀ is 0.12 μg/ml; MIC ₉₀ is 0.5 μg/ml). As a comparison, the clinical combination CEF/VNRX has MIC ₅₀ /MIC ₉₀ values of 1 and 2 μg/ml, respectively. The clinical combination CAZ/AVI is characterized by MIC ₅₀ /MIC ₉₀ values of 2 and 8 μg/ml, respectively.

In FIG. 2 shows data showing the effect of the compound of Example 10 on a panel of clinical strains of KPC-positive enterobacteria resistant to beta-lactam antibiotics such as meropenem, cefepime, and ceftazidime. The MIC data is shown in Fig. 2 in standard cumulative form. In FIG. 2 shows that against KPC-positive enterobacteria, meropenem also shows little activity (MIC ₅₀ greater than 32 μg/ml and MIC90 greater than 32 μg/ml). Addition of the compound of example 10 at a concentration of 4 μg/ml restores susceptibility to meropenem (MIC ₅₀ is 0.12 μg/ml; MIC90 is 2 μg/ml). As a comparison, the clinical combination of CEF/VNRX (VNRX-5133 inhibitor and antibiotic cefepime) has MIC ₅₀ /MIC ₉₀ values of 1 and 2 μg/ml, respectively. The clinical combination of CAZ/AVI (avibactam inhibitor and antibiotic ceftazidime) is characterized by MIC ₅₀ /MIC ₉₀ values of 2 and 4 μg/ml, respectively.

Example 10 was also tested against a panel of 48 strains of A. baumannii resistant to beta-lactam antibiotics such as meropenem, cefepime and ceftazidime because they produce OXA variants of the SBL enzymes. The MIC data is shown in Fig. 3 in standard cumulative form. For example, in the presence of each of the inhibitors at a concentration of 2 μg/ml, 0% of the strains are susceptible to (1) meropenem as monotherapy (MEM), (2) clinical combination of the VNRX-5133 inhibitor and the antibiotic cefepime, CEF/VNRX; or (3) clinical combination of the inhibitor avibactam and the antibiotic ceftazidime, CAZ/AVI. In contrast, approximately 50% are susceptible to the compound of Example 10 in combination with the antibiotic meropenem. Similarly, in the presence of each of the inhibitors at a concentration of 16 μg/ml, less than 10% of the strains are susceptible to (1) meropenem as monotherapy (MEM), (2) clinical combination of the VNRX-5133 inhibitor and the antibiotic cefepime, CEF/VNRX; or (3) clinical combination of the inhibitor avibactam and the antibiotic ceftazidime, CAZ/AVI. On the contrary, under these conditions, almost 100% of the strains are susceptible to the compound of example 10 in combination with the antibiotic meropenem. With respect to MIC values, meropenem was shown to show little activity against this panel (MIC ₅₀ is 64 μg/ml and MIC ₉₀ is greater than 64 μg/ml). Addition of the compound of example 10 at a concentration of 4 μg/ml restores susceptibility to meropenem (MIC ₅₀ is 2 μg/ml; MIC90 is 8 μg/ml). As a comparison, the CEF/VNRX clinical combination showed little activity against these OXA-positive clinical isolates of Acinetobacter baumanni with MIC ₅₀ /MIC ₉₀ values of 64 μg/ml and greater than 64 μg/ml, respectively. Similarly, the CAZ/AVI clinical combination also showed little activity against these OXA-positive clinical isolates of Acinetobacter baumanni, with the combination of the inhibitor avibactam and the antibiotic ceftazidime having MIC ₅₀ /MIC ₉₀ values of 64 µg/mL and greater than 64 µg/mL respectively.

In vivo efficacy of the compounds of the invention

Compounds of the invention were further tested in animal efficacy models. Mice were infected by injection into the thigh of (A) a bovine-expressing clinical strain of Klebsiella pneumoniae (NR-48977) (MIC (meropenem as monotherapy) is 64 μg/ml; MIC (meropenem + compound of example 10 in concentrated

- 52 042632 4 µg/ml) is 1 µg/ml); or (B) OXA-expressing clinical strain of Klebsiella pneumoniae (AC00783) (MIC (meropenem as monotherapy) is 32 µg/ml; MIC (meropenem + compound of example 10 at 4 µg/ml) is 0.25 µg/ml) ; or (C) OXA-expressing clinical strain Acinetobacter baumannii (AC00445) (MIC (meropenem as monotherapy) is 64 μg/ml; MIC (meropenem + compound of example 10 at 4 μg/ml) is 4 μg/ml).

The diluent, meropenem as monotherapy or meropenem and the compound of the invention (from Example 10) were administered iv. 1, 3, 5 and 7 hours after infection. 9 hours after infection, the animals were sacrificed and the number of colony forming units (CFU) was measured to quantify the bacterial load (number of colony forming units per gram of thigh tissue, CFU/g). The results of these experiments, which were performed using appropriate controls and statistical analysis, are shown in FIG. 4. For all three bacterial strains tested, a dose-dependent effect on bacterial load can be seen in the form of lower colony forming units (CFU) when the dose of Example 10 is increased with a constant dose of meropenem.

Claims (23)

1) R is C _1-4 alkyl, wherein the C _1-4 alkyl group is substituted with at least one halogen atom and optionally additionally substituted with one R ² substituent; or 1. A compound that is diazabicyclooctanone of formula (I) or a pharmaceutically acceptable salt thereof, FORMULA (I) where R is selected from halogen, C ₁ . ₄ alkyl and LXR ¹ , while C ₁ . _{the 4} alkyl group is substituted with at least one halogen atom and optionally additionally substituted with one or two R ² substituents; R ¹ is C ₁ . ₄ alkyl which is substituted with at least one halogen atom and optionally additionally substituted with one or two R ² substituents; each R ² is independently a thiazole; L is a bond or is C ₁ . ₂ an alkylene group which is unsubstituted or substituted by at least one halogen atom; And X is O or S(O)z, where z is 0. 2) R ¹ is C _1-2 alkyl, wherein the C _1-2 alkyl group is substituted with at least two halogen atoms selected from fluorine and chlorine atoms; or 2. A compound according to claim 1, wherein said compound is diazabicyclooctanone of formula (II) or a pharmaceutically acceptable salt thereof, or diazabicyclooctanone of formula (III) or a pharmaceutically acceptable salt thereof: Formula (II) Formula (III) where R is halogen. 3) said compound is as defined in claim 5 and R is LXR ¹ where L is a bond or is an unsubstituted C1 alkylene group; X is O or S; And R ¹ is a C1 alkyl group substituted with 1, 2 or 3 halogen groups; or 3. The compound according to claim 1 or 2, where R represents a fluorine or chlorine atom. 4) R is selected from -CH ₂ -O-CF ₃ and -S-CF ₃ ; or 4. The compound according to claim 1, where the specified connection is selected from (2R,5R)-2-fluoro-7-oxo-1,6-diazabicyclo[3.2.1]octan-6-yl-hydrosulfate; (2S,5R)-2-fluoro-7-oxo-1,6-diazabicyclo[3.2.1]octan-6-yl-hydrosulfate; (2R,5R)-2-chloro-7-oxo-1,6-diazabicyclo[3.2.1]octan-6-yl-hydrosulfate; and their pharmaceutically acceptable salts. 5) R is selected from CF ₃ , CHF ₂ , CHCl ₂ , CCl ₃ , CH ₂ F, CF ₂ CH ₃ , CF ₂ -thiαzolyl and CH ₂ CF ₃ , preferably from CF 3 , CHF 2 and CHCl. 5. The compound according to claim 1, which is diazabicyclooctanone of formula (II) or a pharmaceutically acceptable salt thereof, - 53 042632 Ν / )—\ ο ο—SO ₃ H FORMULA (II) where R is as defined in claim 1. 6. The compound according to claim 1 or 5, which is diazabicyclooctanone of formula (II) or a pharmaceutically acceptable salt thereof, -\ O O --- SO ₃ H FORMULA (II) where R is C _1-4 alkyl, wherein the C _1-4 alkyl group is substituted with at least one halogen atom and optionally additionally substituted with one or two R ² substituents; each R ² is independently a thiazole. 7. Connection according to claim 5 or 6, where 8. The compound according to claim 1, which is (2R,5R)-2-fluoro-7-oxo-1,6-diazabicyclo[3.2.1]octan6-yl hydrogen sulfate or a pharmaceutically acceptable salt thereof. 9. The compound according to claim 1, which is (2R,5R)-2-fluoro-7-oxo-1,6-diazabicyclo[3.2.1]octan6-sulfate sodium. 10. A pharmaceutical composition containing a compound according to any one of claims 1 to 9 and a pharmaceutically acceptable carrier or diluent. 11. Pharmaceutical composition according to claim 10, additionally containing a β-lactam antibiotic. 12. The pharmaceutical composition of claim 11 wherein the β-lactam antibiotic is meropenem. 13. A combination of a compound according to any one of claims 1 to 9 and a β-lactam antibiotic for the treatment or prevention of a bacterial infection. 14. The combination of claim 13 wherein the β-lactam antibiotic is meropenem. 15. The use of a compound according to any one of claims 1 to 9, a composition according to any one of claims 10 to 12, or a combination according to claims 13 or 14 to eliminate or reduce antibiotic resistance in Gram-negative bacteria. 16. Use according to claim 15, wherein the Gram-negative bacteria are selected from Enterobacteria, Pseudomonas and Moraxella. 17. Use according to claim 16, wherein the bacteria selected from Enterobacteria, Pseudomonas and Moraxella are selected from Klebsiella pneumoniae, Escherichia coli, Enterobacter cloacae, Pseudomonas aeruginosa, Burkholderia cepacia and Acinetobacter baumannii. 18. The use of a compound according to any one of claims 1 to 9, a composition according to any one of claims 10 to 12 or a combination according to claims 13 or 14 for the treatment or prevention of a bacterial infection. 19. Use according to claim 18, comprising the use of a compound according to any one of claims 1 to 9 for the treatment or prevention of a bacterial infection by co-administration with a β-lactam antibiotic. - 54 042632 20. Use according to claim 19, wherein the β-lactam antibiotic is meropenem. 21. Use according to any one of claims 18-20, wherein the bacterial infection is caused by bacteria selected from Enterobacteria, Pseudomonas and Moraxella. 22. Use according to claim 21, wherein the bacteria selected from Enterobacteria, Pseudomonas and Moraxella are selected from Klebsiella pneumoniae, Escherichia coli, Enterobacter cloacae, Pseudomonas aeruginosa, Burkholderia cepacia and Acinetobacter baumannii. 23. Use according to any one of claims 18-20, wherein the bacterial infection is caused by carbapenem-resistant enterobacteria.