WO2013153394A1 - Quinolonones with antibacterial properties - Google Patents

Abstract

This invention relates to derivatives of fluoroquinolone antibacterial drug compounds. It also relates to pharmaceutical formulations of derivatives of fluoroquinolone antibacterial drug compounds. It also relates to uses of the derivatives in treating bacterial infections and in methods of treating bacterial infections.

Description

QUINOLONONES WITH ANTIBACTERIAL PROPERTIES

This invention relates to derivatives of antibacterial drug compounds. It also relates to pharmaceutical formulations of derivatives of antibacterial drug compounds. It also relates to uses of the derivatives in treating bacterial infections and in methods of treating bacterial infections.

The fluoroquinolone antibacterial family are synthetic broad-spectrum antibiotics. They were originally introduced to treat Gram negative bacterial infections, but are also used for the treatment of Gram positive strains. One problem with existing fluoroquinolones can be the negative side effects that may sometimes occur as a result of fluoroquinolone use. In general, the common side-effects are mild to moderate but, on occasion, more serious adverse effects occur. Some of the serious side effects that occur, and which occur more commonly with fluoroquinolones than with other antibiotic drug classes, include central nervous system (CNS) toxicity and cardiotoxicity. In cases of acute overdose there may be renal failure and seizure.

A particular problem, both with antibiotics in general and with fluoroquinolones in particular, is the increasing frequency of resistant strains of bacterial pathogens such as Staphylococcus aureus, Streptococcus pneumonia, Clostridium difficile and Pseudomonas aeruginosa. In fact, multidrug resistance has become the norm for some pathogens. Of these, Staphylococcus aureus, a Gram positive bacteria, is the most concerning due to its potency and its capacity to adapt to environmental conditions. MRSA (methicillin resistant Staphylococcus aureus) is probably the most well known and has reached pandemic proportions. Of particular concern is the increasing incidence of 'community acquired' infections, those occurring in subjects with no prior hospital exposure. Many strains of MRSA are also resistant to fluoroquinolones, in addition to β-lactam antibiotics such as methicillin.

While less wide-spread, antibiotic resistant Gram negative strains, such as either E. Coli NDM- 1 (New Delhi metallo-p-lactamase) mutation or Klebsiella pneumoniae with the same mutation, are very difficult to treat, with only expensive antibiotics such as vancomycin and colistin being effective.

The present invention seeks to overcome the disadvantages of known fluoroquinolones. In spite of the numerous different antibiotics known in the art for a variety of different infections, there continues to be a need to provide antibiotics that can provide an effective treatment in a reliable manner. In addition, there remains a need for antibiotic drugs which can avoid or reduce the side-effects associated with known antibiotics. A further aim is to provide treatment which is effective in a selective manner at a chosen site of interest. Another aim is to provide antibiotics having a convenient pharmacokinetic profile and a suitable duration of action following dosing.

It is an aim of this invention to provide new antibiotics. In particular it is an aim to provide antibiotics which are active against resistant strains of Gram positive and/or Gram negative bacteria. It is also an aim to provide antibiotics which are active against strains of bacteria which may be associated with biowarfare. Such strains are typified by high mortality and/or rapid onset of disease.

A further aim of the present invention is to provide antibiotics in which the metabolised fragment or fragments of the drug after absorption are GRAS (Generally Regarded As Safe). A further aim of the invention is to provide prodrugs which are not species dependent and/or which reduce inter-patient variability due to differences in metabolism. Another aim of the invention is to provide prodrugs which are able to overcome the food effect in the sense that they can be administered to fed or fasted patients without the need to control carefully the dosing schedule relative to meal times.

The present invention satisfies some or all of the above aims.

According to a first aspect of the present invention, there is provided a compound of formula (I):

X is C or N;

Y is O or NR³;

R¹ is independently selected from the group consisting: H, C_1-4 alkyl or Ac;

R² is independently selected from the group consisting: H, C Ce alkyl, C₂-C₆-alkenyl, C₂-C₆- alkynyl, C Ce haloalkyi, C₃-C₈ cydoalkyi, C₃-C₈ heterocycloalkyl, -(CH₂)n-C₃-C8 cydoalkyi, - (CH₂)n-C3-C₈ heterocycloalkyl, aryl, -(CH₂)_n-aryl, -(CO)-aryl, -(CO)-(CH₂)_n-aryl, heteroaryl, -(CO)- heteroaryl, -(CH₂)_n-heteroaryl and -(CO)-(CH₂)_n-heteroaryl; wherein each n is independently 1 , 2, 3 or 4;

R³ is independently selected at each occurrence from: H, C_1-6 alkyl or Ac; or R² and R³, together with the nitrogen to which they are attached form a 3- to 7- membered ring which optionally contains an O, S or NR⁴ group;

R⁴ is independently at each occurrence H, C_1-6 alkyl or C(0)-CrC₆-alkyl;

R⁵ is selected from the group consisting of: C_rC₄ alkyl, C_rC₄ haloalkyi, C₃-C₅ cydoalkyi, C₃-C₅ halocycloalkyl; unsubstituted phenyl; phenyl substituted with from 1 to 3 independently selected halogen atoms; unsubstituted pyridyl; and pyridyl substituted with from 1 to 3 independently substituents selected from the group consisting of: halo and NHR_a; wherein R_a is H or Ac;

R⁶ is selected from the group consisting of: H, C_rC₄ alkyl and C_rC₄ haloalkyi; or alternatively, R⁵ and R⁶, together with the atoms to which they are attached to form a 4-6- membered ring which optionally contains an O or S atom; wherein the 4-6-membered ring is optionally substituted with 1 or 2 groups independently selected from halo and Ci-C₄ alkyl;

R⁹ is selected from the group consisting: H, NHR^a or C C₄-alkyl; wherein R_a is H or Ac;

R¹⁰ is independently selected from the group: H or F;

R¹¹ is selected from the group consisting of: an N-heterocycloalkyl group and a C₃-C₈ cydoalkyi group; wherein the N-heterocycloalkyl group comprises from 5 to 10 ring atoms and at least one nitrogen atom wherein the N-heterocycloalkyl group is optionally substituted with from 1-3 groups independently selected from halo, tri(C₁-C₄ alkyl)silyloxy, hydroxyl, C C₄ alkyl, oxo or oxime and wherein any nitrogen which does not attach the N-heterocycloalkyl group to the rest of the compound of Formula (I) is an NR^a group; and the C₃-C₈ cydoalkyi group is optionally substituted with at least one NHR^a group and optionally further substituted with from 1-3 groups independently selected from halo, hydroxyl, tri(C₁-C₄ alkyl)silyloxy, C C₄ alkyl, oxo or oxime; wherein R^a is H or Ac;

R¹² is absent or is selected from the group consisting of: H, OR¹⁶ and halo; wherein R¹⁶ is selected from the group consisting of: C C₄ alkyl and C C₄ haloalkyl; or R¹² and R⁵, together with the atoms to which they are attached form a saturated or unsaturated 5- to 7- membered ring which optionally contains an O, S or NR⁴ group; wherein the 5-7-membered ring is optionally substituted with 1 or 2 groups independently selected from halo, C C₄ alkyl and C C₄ haloalkyl; wherein if X is N, R¹² is absent; wherein each of the aforementioned alkyl, haloalkyl, cydoalkyi, halocycloalkyl, aryl (e.g. phenyl) and heteroaryl (e.g. pyridyl) groups are optionally substituted, where chemically possible, by 1 to 3 substituents which are each independently selected at each occurrence from the group consisting of: oxo, imino, oximo, halo, nitro, cyano, hydroxyl, amino, N- heterocycloalkyl, S0₃R^b, S0₂R^b, S0₂NR^bR^bC0₂R^b C(0)R^b, CONR^bR^b, C C₄-alkyl, C C₄ haloalkyi, C C₄ alkoxy, and C C₄ haloalkoxy, wherein R is selected from H, C C₄ alkyl and C C₄ haloalkyi.

In an embodiment, there is provided a compound of formula

; wherein

X is C or N;

Y is O or NR³;

R¹ is independently selected from the group consisting: H, C_1-4 alkyl or Ac;

R² is independently selected from the group consisting: H, C Ce alkyl, C₂-C₆-alkenyl, C₂-C₆- alkynyl, C Ce haloalkyi, C₃-C₈ cycloalkyi, C₃-C₈ heterocycloalkyl, -(CH₂)n-C₃-C8 cycloalkyi, - (CH₂)n-C₃-C8 heterocycloalkyl, aryl, -(CH₂)_n-aryl, -(CO)-aryl, heteroaryl, -(CO)-heteroaryl, and - (CH₂)n-heteroaryl; wherein each n is independently 1 , 2, 3 or 4;

R³ is independently selected at each occurrence from: H, C_1-6 alkyl or Ac; or R² and R³, together with the nitrogen to which they are attached form a 3- to 7- membered ring which optionally contains an O, S or NR⁴ group;

R⁴ is independently at each occurrence H, C_1-6 alkyl or C(0)-CrC₆-alkyl;

R⁵ is selected from the group consisting of: C C₄ alkyl, C C₄ haloalkyi, C₃-C₅ cycloalkyi, C₃-C₅ halocycloalkyl; unsubstituted phenyl; phenyl substituted with from 1 to 3 independently selected halogen atoms; unsubstituted pyridyl; and pyridyl substituted with from 1 to 3 independently substituents selected from the group consisting of: halo and NHR_a; wherein R^a is H or Ac;

R⁶ is selected from the group consisting of: H, C C₄ alkyl and C C₄ haloalkyl;

or alternatively, R⁵ and R⁶, together with the atoms to which they are attached to form a 4-6- membered ring which optionally contains an O or S atom; wherein the 4-6-membered ring is optionally substituted with 1 or 2 groups independently selected from halo and C C₄ alkyl;

R⁹ is selected from the group consisting: H, NHR^a or CrC₄-alkyl; wherein R^a is H or Ac;

R¹⁰ is independently selected from the group: H or F;

R¹¹ is selected from the group consisting of: an N-heterocycloalkyl group and a C₃-C₈ cydoalkyi group; wherein the N-heterocycloalkyl group comprises from 5 to 10 ring atoms and at least one nitrogen atom wherein the N-heterocycloalkyl group is optionally substituted with from 1-3 groups independently selected from halo, tri(C₁-C₄ alkyl)silyloxy, hydroxyl, C C₄ alkyl, oxo or oxime and wherein any nitrogen which does not attach the N-heterocycloalkyl group to the rest of the compound of Formula (I) is an NR^a group; and the C₃-C₈ cydoalkyi group is optionally substituted with at least one NHR^a group and optionally further substituted with from 1-3 groups independently selected from halo, hydroxyl, tri^-C^ alkyl)silyloxy, C C₄ alkyl, oxo or oxime; wherein R_a is H or Ac;

R¹² is absent or is selected from the group consisting of: H, OR¹⁶ and halo; wherein R¹⁶ is selected from the group consisting of: C C₄ alkyl and C C₄ haloalkyl; wherein if X is N, R¹² is absent; wherein each of the aforementioned alkyl, haloalkyl, cydoalkyi, halocycloalkyl, aryl (e.g. phenyl) and heteroaryl (e.g. pyridyl) groups are optionally substituted, where chemically possible, by 1 to 3 substituents which are each independently selected at each occurrence from the group consisting of: oxo, imino, oximo, halo, nitro, cyano, hydroxyl, amino, C0₂H, C0₂-(C₁-C₄alkyl), C(0)H, C C₄-alkyl, C C₄ haloalkyl, C C₄ alkoxy, and C C₄ haloalkoxy. In an embodiment, the compound of formula (I) is a compound of formula (II):

R¹² and X are as described above.

In an embodiment, the compound of formula (I) is a compound of formula

X are as described above.

In an embodiment, there is provided a derivative of an antibacterial compound selected from the group comprising: enoxacin, fleroxacin, lomefloxacin, nadifloxacin, norfloxacin, rufloxacin, balofloxacin, grepafloxacin, pazufloxacin, sparfloxacin, temafloxacin, tosufloxacin, besifloxacin, clinafloxacin, garenoxacin, gemifloxacin, gatifloxacin, sitafloxacin, trovafloxacin, prulifloxacin, ciprofloxacin, pefloxacin, moxifloxacin, ofloxacin, levofloxacin, delafloxacin and jnj-q2, in which the carboxylic acid of the antibacterial compound is replaced by a group having the following partial formula (IV):

wherein:

Y is O or NR³;

R¹ is independently selected from the group consisting: H, C_1-4 alkyl or Ac;

R² is independently selected from the group consisting: C Ce alkyl, C₂-C₆-alkenyl, C₂-C₆- alkynyl, Ci-C₆ haloalkyl, C₃-C₈ cycloalkyl, C₃-C₈ heterocycloalkyl, -(CH₂)n-C₃-C8 cycloalkyl, - (CH₂)n-C₃-C8 heterocycloalkyl, aryl, -(CH₂)_n-aryl, -(CO)-aryl, heteroaryl -(CO)-heteroaryl, and - (CH₂)n-heteroaryl; wherein each n is independently 1 , 2, 3 or 4;

R³ is independently selected at each occurrence from: H, C_1-6 alkyl or Ac; or R² and R³, together with the nitrogen to which they are attached form a 3- to 7- membered ring which optionally contains an O, S or NR⁴ group;

R⁴ is independently at each occurrence H, C_1-6 alkyl or C^-C Ce-alkyl; wherein each of the aforementioned alkyl, haloalkyl, cycloalkyl, halocycloalkyl, aryl (e.g. phenyl) and heteroaryl (e.g. pyridyl) groups are optionally substituted, where chemically possible, by 1 to 3 substituents which are each independently selected at each occurrence from the group consisting of: oxo, imino, oximo, halo, nitro, cyano, hydroxyl, amino, C0₂H, C0₂-(C₁-C₄alkyl), C(0)H, d-C-alkyl, C C₄ haloalkyl, C C₄ alkoxy, and C C₄ haloalkoxy; and wherein, if present, any one or more amine group in the fluoroquinolone may optionally be acetylated.

In an embodiment, the antibacterial compound is selected from: fleroxacin, lomefloxacin, nadifloxacin, norfloxacin, rufloxacin, balofloxacin, grepafloxacin, pazufloxacin, sparfloxacin, temafloxacin, tosufloxacin, besifloxacin, clinafloxacin, garenoxacin, gemifloxacin, gatifloxacin, sitafloxacin, trovafloxacin, prulifloxacin, ciprofloxacin, pefloxacin, moxifloxacin, delafloxacin and jnj-q2 In an embodiment, one amine group in the fluoroquinolone is acetylated.

In another aspect of the present invention, there is provided a method of preparing a formulation of a derivative of an antibacterial compound as defined above, the method comprising:

(i) obtaining the derivative of an antibacterial drug molecule; and

(ii) mixing the derivative with one or more pharmaceutically acceptable excipients to produce the pharmaceutical formulation.

Synthetic procedures for the formation of amides are well known in the art.

The compounds of the invention are based on the parent approved pharmaceutically active compounds disclosed below. The synthetic routes to each of the compounds are available in the literature and in the relevant EMA and FDA regulatory files and accordingly are not reproduced here. These disclosures insofar as the synthetic procedures are concerned form part of the disclosure of the present invention. In the interests of brevity, the details of these synthetic procedures are not reproduced here but it is intended that this subject matter is specifically incorporated into the disclosure of these documents by reference. rs to a fluoroquinolone with the following structure:

The skilled man will appreciate that adaptation of methods known in the art could be applied in the manufacture of the compounds of the present invention.

For example, the skilled person will be immediately familiar with standard textbooks such as "Comprehensive Organic Transformations - A Guide to Functional Group Transformations", RC Larock, Wiley-VCH (1999 or later editions), "March's Advanced Organic Chemistry - Reactions, Mechanisms and Structure", MB Smith, J. March, Wiley, (5th edition or later) "Advanced Organic Chemistry, Part B, Reactions and Synthesis", FA Carey, RJ Sundberg, Kluwer Academic/Plenum Publications, (2001 or later editions), "Organic Synthesis - The

Disconnection Approach", S Warren (Wiley), (1982 or later editions), "Designing Organic Syntheses" S Warren (Wiley) (1983 or later editions), "Guidebook To Organic Synthesis" RK Mackie and DM Smith (Longman) (1982 or later editions), etc., and the references therein as a guide.

The skilled chemist will exercise his judgement and skill as to the most efficient sequence of reactions for synthesis of a given target compound and will employ protecting groups as necessary. This will depend inter alia on factors such as the nature of other functional groups present in a particular substrate. Clearly, the type of chemistry involved will influence the choice of reagent that is used in the said synthetic steps, the need, and type, of protecting groups that are employed, and the sequence for accomplishing the protection / deprotection steps. These and other reaction parameters will be evident to the skilled person by reference to standard textbooks and to the examples provided herein.

Sensitive functional groups may need to be protected and deprotected during synthesis of a compound of the invention. This may be achieved by conventional methods, for example as described in "Protective Groups in Organic Synthesis" by TW Greene and PGM Wuts, John Wiley & Sons Inc (1999), and references therein.

Each of the compounds of the present invention may be used as a medicament. Thus, in another aspect of the invention, there is provided a derivative of an antibacterial compound as defined in this specification for the treatment of antibacterial infections.

The invention includes phamaceutical formulations comprising a compound of the invention and optionally, a pharmaceutically acceptable exceipient. Such formulations may also include other active agents.

The compounds and formulations of the present invention may be used in the treatment of a wide range of bacterial infections. In some embodiments, the compounds can be used to treat bacterial infections caused by one or more resistant strains of bacteria. In a further embodiment, the compounds can be used to treat bacterial infections caused by one or more resistant strains of Gram positive bacteria. In a further embodiment, the compounds can be used to treat bacterial infections caused by one or more resistant strains of Gram negative bacteria.

The term 'resistant strains' is intended to mean strains of bacteria which have shown resistance to one or more known antibacterial drug. For example, it may refer to strains which are resistant to meticillin, strains that are resistant to other β-lactam antibiotics and/or strains that are resistant to fluoroquinolones which do not fall within this application. A resistant strain is one in which the MIC 100 of a given compound or class of compounds for that strain has shifted to a significantly higher number than for the parent (susceptible) strain.

The compounds and formulations of the present invention can be used to treat or to prevent infections caused by bacterial strains associated with biowarfare. These may be strains which are category A pathogens as identified by the US government (e.g. those which cause anthrax, plague etc.) and/or they may be strains which are category B pathogens as identified by the US government (e.g. those which cause Glanders disease, mellioidosis etc). In a specific embodiment, the compounds and formulations of the present invention can be used to treat or to prevent infections caused by Gram positive bacterial strains associated with biowarfare (e.g. anthrax). More particularly, the compounds and formulations may be used to treat category A and/or category B pathogens as defined by the US government on 1 ^st April 2013.

The compounds and formulations of the present invention can be used to treat both Gram positive and Gram negative bacterial infections such as infections of the urinary tract, the respiratory tract, the ear, the skin, the throat, soft tissue, bone and joints (including infections caused by Staph Aureus). The compounds can be used to treat pneumonia, sinusitis, acute bacterial sinusitis, bronchitis, acute bacterial exacerbation of chronic bronchitis, anthrax, chronic bacterial prostatitis, acute pyelonephritis, pharyngitis, tonsillitis, eColi, prophylaxis before dental surgery, cellulitis, acnes, cystitis, infectious diarrhoea, typhoid fever, infections caused by anaerobic bacteria, peritonitis, malaria, babesiosis bacterial vaginosis, pelvic inflammatory disease, pseudomembranous colitis, helicobacter pylori, amoebiasis, giardasis, acute gingivitis, Crohn's Disease, rosacea, fungating Tumours, MRSA, impetigo. The compounds of the present invention may also be used in treating other conditions treatable by eliminating or reducing a bacterial infection. In this case they will act in a secondary manner alongside for example a chemotherapeutic agent used in the treatment of cancer.

The compounds of the present invention can be used in the treatment of the human body. They may be used in the treatment of the animal body. In particular, the compounds of the present invention can be used to treat commercial animals such as livestock. Alternatively, the compounds of the present invention can be used to treat companion animals such as cats, dogs, etc.

The parent compound upon which the derivatives of the invention are based may be selected from the group consisting of: Enoxacin, Fleroxacin, Lomefloxacin, Nadifloxacin, Norfloxacin, Rufloxacin, Balofloxacin, Grepafloxacin, Pazufloxacin, Sparfloxacin, Temafloxacin, Tosufloxacin, Besifloxacin, Clinafloxacin, Garenoxacin, Gemifloxacin, Gatifloxacin, Sitafloxacin, Trovafloxacin, Prulifloxacin, Ciprofloxacin, Pefloxacin, Moxifloxacin, Ofloxacin, jnj- q2, Levofloxacin and Delafloxacin.

Compounds of the invention intended for pharmaceutical use may be administered as crystalline or amorphous products. They may be obtained, for example, as solid plugs, powders, or films by methods such as precipitation, crystallization, freeze drying, or spray drying, or evaporative drying. Microwave or radio frequency drying may be used for this purpose.

Compounds of the invention containing one or more asymmetric carbon atoms can exist as two or more stereoisomers. Where a compound of the invention contains a double bond such as a C=C or C=N group, geometric cis/trans (or Z E) isomers are possible. Specifically, any oxime group present in the compounds of the invention may be present as the E-oxime, as the Z-oxime or as a mixture of both in any proprotion. Where structural isomers are interconvertible via a low energy barrier, tautomeric isomerism ('tautomerism') can occur. This can take the form of proton tautomerism in compounds of the invention containing, for example, an imino, keto, or oxime group, or so-called valence tautomerism in compounds which contain an aromatic moiety. It follows that a single compound may exhibit more than one type of isomerism. Included within the scope of the present invention are all stereoisomers, geometric isomers and tautomeric forms of the compounds of the invention, including compounds exhibiting more than one type of isomerism, and mixtures of one or more thereof. Also included are acid addition or base salts wherein the counter ion is optically active, for example, d-lactate or l-lysine, or racemic, for example, dl-tartrate or dl-arginine.

Cis/trans isomers may be separated by conventional techniques well known to those skilled in the art, for example, chromatography and fractional crystallisation.

Conventional techniques for the preparation/isolation of individual enantiomers when necessary include chiral synthesis from a suitable optically pure precursor or resolution of the racemate (or the racemate of a salt or derivative) using, for example, chiral high pressure liquid chromatography (HPLC).

Alternatively, the racemate (or a racemic precursor) may be reacted with a suitable optically active compound, for example, an alcohol, or, in the case where the compound of the invention contains an acidic or basic moiety, a base or acid such as 1-phenylethylamine or tartaric acid. The resulting diastereomeric mixture may be separated by chromatography and/or fractional crystallization and one or both of the diastereoisomers converted to the corresponding pure enantiomer(s) by means well known to a skilled person.

Chiral compounds of the invention (and chiral precursors thereof) may be obtained in enantiomerically-enriched form using chromatography, typically HPLC, on an asymmetric resin with a mobile phase consisting of a hydrocarbon, typically heptane or hexane, containing from 0 to 50% by volume of isopropanol, typically from 2% to 20%, and from 0 to 5% by volume of an alkylamine, typically 0.1 % diethylamine. Concentration of the eluate affords the enriched mixture.

When any racemate crystallises, crystals of two different types are possible. The first type is the racemic compound (true racemate) referred to above wherein one homogeneous form of crystal is produced containing both enantiomers in equimolar amounts. The second type is the racemic mixture or conglomerate wherein two forms of crystal are produced in equimolar amounts each comprising a single enantiomer. While both of the crystal forms present in a racemic mixture have identical physical properties, they may have different physical properties compared to the true racemate. Racemic mixtures may be separated by conventional techniques known to those skilled in the art - see, for example, "Stereochemistry of Organic Compounds" by E. L. Eliel and S. H. Wilen (Wiley, 1994).

In an embodiment, the present invention provides a compound according to any one or more than one of the following formulae 1-27 taken alone or in any combination:





wherein:

Y is O or NR ;

R¹ is independently selected from the group consisting: H , Ci _-4 alkyl or Ac;

R² is independently selected from the group consisting: C Ce alkyl, C₂-C₆-alkenyl, C₂-C₆- alkynyl, C Ce haloalkyl, C₃-C₈ cycloalkyl, C₃-C₈ heterocycloalkyl, -(CH₂)n-C₃-C8 cycloalkyl, - (CH₂)n-C₃-C8 heterocycloalkyl, aryl, -(CH₂)_n-aryl, -(CO)-aryl, heteroaryl, -(CH₂)_n-heteroaryl and - (CO)-heteroaryl; wherein each n is independently 1 , 2, 3 or 4;

R³ is independently selected at each occurrence from: H , Ci _-6 alkyl or Ac; or R² and R³, together with the nitrogen to which they are attached form a 3- to 7- membered ring which optionally contains an O, S or N R⁴ group;

R⁴ is independently at each occurrence H, C_1-6 alkyl or C(0)-CrC₆-alkyl; wherein each of the aforementioned alkyl, haloalkyl, cycloalkyl, halocycloalkyl, aryl (e.g. phenyl) and heteroaryl (e.g. pyridyl) groups are optionally substituted, where chemically possible, by 1 to 3 substituents which are each independently selected at each occurrence from the group consisting of: oxo, imino, oximo, halo, nitro, cyano, hydroxyl, amino, C0₂H, C0₂-(C₁-C₄alkyl), C(0)H, d-C-alkyl, C C₄ haloalkyl, C C₄ alkoxy, and C C₄ haloalkoxy;

R_a is independently at each occurrence H or Ac.

The compound may be selected from one, some or all of the group of compounds defined by formulae 1-27. Thus, it may be selected from a smaller group such as that defined by a single formula from within the formulae 1 to 27 (e.g. any one of formulae 4, 21 , 22, 23, 24 and 27), or from a group of compounds defined by more than one (e.g. from two to twenty) of any of the above formulae (e.g. the group defined by formulae 4, 21 , 22, 23, 24 and 27) taken together or in combination.

In an embodiment, the compound is not a compound of formula 1 , formula 26 or formula 27.

The following embodiments apply to any of the embodiments of the invention previously described, as appropriate:

In an embodiment R¹ is H.

In an embodiment, Y is NR³. In an alternative embodiment, Y is O.

In an embodiment, R² is selected from the group consisting: Ci-C₆ alkyl, C₂-C₆-alkenyl, C₂-C₆- alkynyl, CrC₆ haloalkyl, C₃-C₈ cycloalkyl, C₃-C₈ heterocycloalkyl, -(CH₂)n-C₃-C8 cycloalkyl, - (CH₂)n-C₃-C8 heterocycloalkyl, aryl, -(CH₂)_n-aryl, -(CO)-aryl, heteroaryl, -(CO)-heteroaryl, and - (CH₂)n-heteroaryl; wherein each n is independently 1 , 2, 3 or 4. In an alternative embodiment, R² is H.

In an embodiment, R² is independently selected from the group consisting: H, Ci-C₆ alkyl, C₂- C₆-alkenyl, C₂-C₆-alkynyl, CrC₆ haloalkyl, C₃-C₈ cycloalkyl, C₃-C₈ heterocycloalkyl, -(CH₂)_n-C₃- C₈ cycloalkyl, -(CH₂)_n-C₃-C₈ heterocycloalkyl, aryl, -(CH₂)_n-aryl, heteroaryl, and -(CH₂)_n- heteroaryl; wherein each n is independently 1 , 2, 3 or 4;

In an embodiment, R² is selected from: CrC₆ alkyl, C₂-C₆-alkenyl, C₂-C₆-alkynyl, CrC₆ haloalkyl, C₃-C₅ cycloalkyl, aryl, -(CH₂)_n-aryl, heteroaryl, -(CO)-heteroaryl and -(CH₂)_n- heteroaryl. In a further embodiment, R² is selected from: Ci-C₆ alkyl, C₂-C₆-alkenyl, Ci-C₆ haloalkyl, -(CO)-heteroaryl, aryl, -(CH₂)_n-aryl and heteroaryl. In yet another embodiment, R² is selected from: C Ce alkyl, C₂-C₆-alkenyl, CrC₆ haloalkyl, aryl, -(CH₂)_n-aryl and heteroaryl.

In an embodiment, R² is C Ce alkyl. In an embodiment, R² is CrC₄ alkyl. Thus, R² may be methyl, ethyl, iso-propyl, n-propyl, iso-butyl, n-butyl, tert-butyl. In a particular embodiment, R² is ethyl (e.g. unsubstituted ethyl, 2-cyanoeth-1-yl or 2-hydroxyeth-1 -yl). In another, R² is tert- butyl. R² may be

In an embodiment, R² is C₂-C₆ alkenyl. In an embodiment, R² is C₃-C₆ alkenyl. Thus, R² may be ally I .

In an alternative embodiment, R² is C Ce haloalkyl. In an embodiment, R² is C C₄ haloalkyl. Thus, R² may be trifluoromethyl or R² may be 1 , 1 , 1 -trifluoroeth-2-yl. In a particular embodiment, R² is 1 , 1 , 1-trifluoroeth-2-yl.

In an alternative embodiment, R² is selected from -(CO)-heteroaryl, -(CO)-aryl -(CO)-(CH₂)_n- aryl, -(CO)-(CH₂)_n-heteroaryl and -(CO^C Cealkyl. In an embodiment, R² is -(CO)-heteroaryl. In a further embodiment, R² is -(CO)-pyridyl, e.g. -(CO)-3-pyridyl.

In a further alternative, R² is aryl or -(CH₂)_n-aryl. Thus, R² may be aryl, e.g. a phenyl group optionally substituted with from 1 to 5 groups independently selected at each occurrence from: halo, nitro, cyano, hydroxyl, amino, N-heterocycloalkyl, S0₃R^B, S0₂R^B, S0₂NR^BR^B C0₂R^B C(0)R^B, CON R^BR^B, C C₄-alkyl, C C₄ haloalkyl, C C₄ alkoxy, and C C₄ haloalkoxy. It may be that the substituent groups are independently selected from: halo; nitro; cyano; hydroxyl; NR⁴R⁴; C0₂H ; C(0)NR⁴R⁴; C0₂-(C C₆alkyl); C(0)H; C C₆-alkyl; C C₄ haloalkyl; C C₄ alkoxy; and Ci-C₆ haloalkoxy. In an embodiment, R² is unsubstituted phenyl. In an alternative embodiment, R² is substituted phenyl. Thus, it may be that R² is not an unsubstituted phenyl ring. Thus, R² may be mono substituted or it may be disubstituted or it may be trisubstituted. In embodiments in which the phenyl is substituted, it may be substituted with at least one electron-withdrawing group. In embodiments in which the phenyl is substituted, it may be substituted with at least 1 group selected from the group consisting of: halo, nitro, S0₃R^B, S0₂R^B, S0₂N R^BR^B C0₂R^B C(0)R^B, CON R^BR^B, cyano, and C C₄ haloalkyl. Thus, R² may be 2,6-dichloro-3-trifluoromethylphenyl. R² may be a phenyl group substituted with one group selected from the group consisting of: halo, nitro, cyano, CO^CrCealkyl), and C C₄ haloalkyl. R² may be a halophenyl group. Thus, R² may be a chlorophenyl group, e.g. a 2-chlorophenyl or a 3-chlorophenyl or a 4-chlorophenyl group. R² may be a fluorophenyl group, e.g. a 2- fluorophenyl or a 3-fluorophenyl or 4-fluorophenyl group. R² may be a bromophenyl group, e.g. a 2-bromophenyl or a 3-bromophenyl or 4-bromophenyl group. In an embodiment, R² may be a nitrophenyl group, e.g. a 2-nitrophenyl or a 3- nitrophenyl or a 4- nitrophenyl group.

In an embodiment, R² may be a para- or meta- substituted phenyl group. This embodiment applies equally to monosubstituted rings, to bisubstituted rings and to trisubstituted phenyl rings. In other words, it may be that if R² is a substituted phenyl group it is not ortho substituted. In a particular embodiment, R² may be a mono- para- or meta substituted phenyl group. The single substituent may be an electron withdrawing group e.g. a group selected from: halo, nitro, S0₃R^b, S0₂R^b, S0₂NR^bR^b C0₂R^b C(0)R^b, CONR^bR^b, cyano, and C C₄ haloalkyl. Thus, the single substituent may be any one of or may be selected from a group consisting of any more than one of: C0₂H, S0₃H, S0₂Me, CN, S0₂NH₂, F, CI, Br, N0₂, OCF₃, CF₃, CONMe₂, CONHMe, CONH₂. The single substituent may also be any one of or may be selected from a group consisting of any more than one of: methyl, OMe, isopropyl, f-butyl.

We have found that in some cases the presence of a meta- or para- substituent has a beneficial effect on the activity.

R² may also be -(CH₂)_n-aryl. The integer n may be 1 or 2, e.g. 1 . Thus, R² may be benzyl in which the aryl ring is optionally substituted with from 1 to 5 groups independently selected at each occurrence from: halo; nitro; cyano; hydroxyl; NR⁴R⁴; C0₂H; C(0)NR⁴R⁴; C0₂-(C C₆alkyl); C(0)H; C C₆-alkyl; C C₄ haloalkyl; C C₄ alkoxy; and C C₆ haloalkoxy. In a particular embodiment, R² is benzyl. In another particular embodiment, R² is pentafluorobenzyl. R2 may be fluorobenzyl (e.g. 4-fluorophenyl)

In another embodiment, R² is heteroaryl or C^Ce alkylheteroaryl. Thus, R² may be a heteroaryl group, e.g. a monocyclic heteroaryl group. In particular, R² may be a 6-membered heteroaryl group containing 1-3-nitrogen atoms. In an embodiment, R² may be pyridine, pyridazine, pyrimidine or pyrazine. Thus, R² may be pyridine (e.g. 2-pyridine) or R² may be pyrimidine (e.g. 2-pyrimidine). In all of these embodiments, the heteroaryl ring is optionally substituted with from 1 to 5 groups independently selected at each occurrence from: halo, nitro, cyano, hydroxyl, amino, N-heterocycloalkyl, S0₃R^b, S0₂R^b, S0₂NR^bR^b C0₂R^b C(0)R^b, CONR^bR^b, C_r C₄-alkyl, C C₄ haloalkyl, C C₄ alkoxy, and C C₄ haloalkoxy. It may be that the substituent groups are independently selected from: halo; nitro; cyano; hydroxyl; NR⁴R⁴; C0₂H; C(0)NR⁴R⁴; C0₂-(C₁-C₆alkyl); C(0)H; C C₆-alkyl; C C₄ haloalkyl; C C₄ alkoxy; and C C₆ haloalkoxy. Alternatively, the heteroaryl may be unsubstituted.

In an embodiment, R² and R³ together with the nitrogen to which they are attached form a 3- to 7- membered ring which optionally contains an O, S or NR⁴ group. In a further embodiment, R² and R³ together with the nitrogen to which they are attached form a 6- membered ring which optionally contains an O, S or NR⁴ group. In a further embodiment, R² and R³ together with the nitrogen to which they are attached form a 6- membered ring, e.g. a piperidine ring. In all of these embodiments, the heterocyclic ring formed by R², R³ and the nitrogen to which they are attached is optionally substituted with from 1 to 5 groups independently selected at each occurrence from: halo; nitro; cyano; hydroxyl; NR⁴R⁴; C0₂H; C(0)NR⁴R⁴; CC CCrCealkyl); C(0)H; C Ce-alkyl; C C₄ haloalkyl; C C₄ alkoxy; and C Ce haloalkoxy. Alternatively, the ring may be unsubstituted.

In an embodiment, it is not the case that R² and R³ together with the nitrogen to which they are attached form a 3- to 7- membered ring which optionally contains an O, S or NR⁴ group.

In an embodiment, R³ is H.

In an embodiment, each R⁴ is H.

In an embodiment X is N. In an alternative embodiment, X is C.

In an embodiment R⁹ is H. Alternatively, R⁹ is NHR_a. In a further alternative embodiment, R⁹ is C C₄ alkyl, e.g. R⁹ may be methyl, ethyl, propyl, butyl, iso-propyl or tert-butyl. In a particular embodiment, R⁹ is methyl.

In an embodiment, R¹⁰ is H. In an alternative embodiment R¹⁰ is F.

In an embodiment, R¹¹ is a N-heterocycloalkyl group comprising from 5 to 10 ring atoms wherein at least one of the ring atoms is an NR_a group wherein the N-heterocycloalkyl group is optionally substituted with from 1 -3 groups independently selected from halo, hydroxyl, C C₄ alkyl, oxo, oxime comprising from 5 to 10 ring atoms and at least one nitrogen atom which is optionally substituted with from 1 -3 groups independently selected from halo, hydroxyl, C C₄ alkyl, oxo, oxime. In an embodiment, the N-heterocycloalkyl group is attached to the remainder of the compound of Formula (I) via the N-atom.

In an embodiment, R¹¹ is a piperazine ring which is optionally substituted with from 1-3 groups independently selected from halo, hydroxyl, C C₄ alkyl, oxo or oxime. In a preferred embodiment, R¹¹ is a piperazine ring which is optionally substituted with from 1-3 independently selected C C₄ alkyl groups. In a further preferred embodiment, R¹¹ is a piperazine ring substituted with a methyl group, e.g. an N-methyl piperazine ring. In an alternative preferred embodiment, R¹¹ is a 3-methyl piperazine ring. In a further alternative, R¹¹ may be a 2-methyl piperazine ring. In another preferred embodiment, R¹¹ is a piperazine ring which is optionally substituted with a C_1-4 alkanoyl group. In an embodiment, the R¹¹ group is an acetyl piperazine group.

In an alternative embodiment, R¹¹ is a piperidine ring which is optionally substituted with from 1-3 groups independently selected from halo, hydroxyl, Ci-C₄ alkyl, oxo, oxime.

In an alternative embodiment,

In an embodiment, the NH moiety in the structure depicted may alternatively be substituted with an acetyl group.

In an alternative embodiment, R¹¹ is C₃-C₈ cycloalkyl group substituted with at least one NHR_a group and optionally further substituted with from 1 -3 groups independently selected from halo, hydroxyl, C C₄ alkyl, oxo or oxime. In a preferred embodiment, R¹¹ is a cyclopropyl group substituted with a NHR_a group.

In an embodiment, R¹² is H. In an alternative embodiment, R¹² is CI or F. In a further alternative, R¹² is OR¹⁶. In an embodiment, R¹⁶ is C C₄ alkyl, e.g. R¹⁶ may be methyl. In an alternative embodiment, R¹⁶ is C C₄ haloalky, e.g. C C₄ fluoroalkyl such as trifluoromethyl or difluoromethyl.

In an alternative embodiment, R¹² and R⁵, together with the atoms to which they are attached, form a 6-membered ring which optionally contains an O or S atom; wherein the 6-membered ring is optionally substituted with 1 or 2 groups independently selected from halo and C C₄ alkyl. In an embodiment, the ring contains an O atom. Alternatively, the ring contains a S atom. The ring may be substituted with a Ci-C₄ alkyl group, e.g. a methyl group.

In an embodiment, R⁵ is C C₄ alkyl, e.g. ethyl. In an embodiment, alternative embodiment R⁵ is C C₄ haloalkyi, e.g. a 2-fluoroethyl group. In a further alternative embodiment, R⁵ is C₃-C₅ cycloalkyl, e.g. cyclopropyl. In yet a further alternative embodiment, R⁵ is C₃-C₅ halocycloalkyl, e.g.fluoro cyclopropyl. In other embodiments R⁵ is phenyl which may be optionally substituted with from 1-3 halo groups, i.e. R⁵ may be a difluorophenyl group, such as 2,4-difluorophenyl.

In an embodiment, R⁶ is H.

In an embodiment, the compound of formula (I) is not a compound selected from:

In another aspect the present invention provides a pharmaceutical formulation comprising a compound of the invention and a pharmaceutically acceptable excipient.

Aryl groups may be 6-membered aryl groups. Aryl groups may be optionally substituted phenyl groups, optionally substituted biphenyl groups, optionally substituted naphthalenyl groups or optionally substituted anthracenyl groups.

Heteroaryl groups may be 5- or 6-membered heteroaryl groups. They may be monocyclic heteroaryl groups, (e.g. heteroaryl groups may be selected from: 5-membered heteroaryl groups in which the heteroaromatic ring is substituted with 1 -3 heteroatoms selected from O, S and N; and 6-membered heteroaryl groups in which the heteroaromatic ring is substituted with 1-2 nitrogen atoms) or they may be bicyclic heteroaryl groups (9-membered bicyclic heteroaryl groups in which the heteroaromatic system is substituted with 1 -4 heteroatoms selected from O, S and N; 10-membered bicyclic heteroaryl groups in which the heteroaromatic system is substituted with 1-4 nitrogen atoms). Specifically, heteroaryl groups may be selected from: pyrrole, furan, thiophene, pyrazole, imidazole, oxazole, isoxazole, triazole, oxadiazole, thiodiazole, pyridine, pyridazine, pyrimidine, pyrazine, triazine (e.g. 1 ,3,5-triazine or 1 ,3,4 triazine), indole, isoindole, benzofuran, isobenzofuran, benzothiophene, indazole,

benzimidazole, benzoxazole, benzthiazole, benzisoxazole, purine, quinoline, isoquinoline, cinnoline, quinazoline, quinoxaline, pteridine, phthalazine, naphthyridine.

Heterocycloalkyi groups of the invention may be from 3-8 membered saturated rings in which the ring contains from 1 to 3 heteroatoms (e.g. O, N or S). Where the heterocycloalkyi group contains a nitrogen, the heterocycloalkyi group may be linked to the remainder of the molecule via the nitrogen or via one of the carbon atoms in the ring. Where the heterocycloalkyi group does not contain a nitrogen, the heterocycloalkyi group will be linked to the rest of the molecule via one of the carbon atoms in the ring.

The aryl and heteroaryl groups are optionally substituted with from 1 to 4 groups independently selected at each occurrence from: halo, nitro, cyano, hydroxyl, NHR_a, C0₂H, C0₂-(CrC₄alkyl), C(0)H, C C4-alkyl, C C₄ haloalkyl, C C₄ alkoxy, and C C₄ haloalkoxy.

The present invention also includes the synthesis of all pharmaceutically acceptable isotopically-labelled compounds of the invnetion wherein one or more atoms are replaced by atoms having the same atomic number, but an atomic mass or mass number different from the atomic mass or mass number usually found in nature.

For the above-mentioned compounds of the invention the dosage administered will, of course, vary with the compound employed, the mode of administration, the treatment desired and the disorder indicated. For example, if the compound of the invention is administered orally, then the daily dosage of the compound of the invention may be in the range from 0.01 micrograms per kilogram body weight ^g/kg) to 100 milligrams per kilogram body weight (mg/kg).

A compound of the invention, or pharmaceutically acceptable salt thereof, may be used on their own but will generally be administered in the form of a pharmaceutical composition in which the compounds of the invention, or pharmaceutically acceptable salt thereof, is in association with a pharmaceutically acceptable adjuvant, diluent or carrier. Conventional procedures for the selection and preparation of suitable pharmaceutical formulations are described in, for example, "Pharmaceuticals - The Science of Dosage Form Designs", M. E. Aulton, Churchill Livingstone, 1988. Depending on the mode of administration of the compounds of the invention, the pharmaceutical composition which is used to administer the compounds of the invention will preferably comprise from 0.05 to 99 %w (per cent by weight) compounds of the invention, more preferably from 0.05 to 80 %w compounds of the invention, still more preferably from 0.10 to 70 %w compounds of the invention, and even more preferably from 0.10 to 50 %w compounds of the invention, all percentages by weight being based on total composition.

The pharmaceutical compositions may be administered topically (e.g. to the skin) in the form, e.g., of creams, gels, lotions, solutions, suspensions, or systemically, e.g. by oral

administration in the form of tablets, capsules, syrups, powders or granules; or by parenteral administration in the form of a sterile solution, suspension or emulsion for injection (including intravenous, subcutaneous, intramuscular, intravascular or infusion); or by rectal administration in the form of suppositories.

If administered topically high-dosages of the compounds of the invention can be administered. Thus, a compound with an in vitro MIC100 of, for example, 16-64 may still provide an effective treatment against bacteria.

For oral administration the compounds of the invention may be admixed with an adjuvant or a carrier, for example, lactose, saccharose, sorbitol, mannitol; a starch, for example, potato starch, corn starch or amylopectin; a cellulose derivative; a binder, for example, gelatine or polyvinylpyrrolidone; and/or a lubricant, for example, magnesium stearate, calcium stearate, polyethylene glycol, a wax, paraffin, and the like, and then compressed into tablets. If coated tablets are required, the cores, prepared as described above, may be coated with a concentrated sugar solution which may contain, for example, gum arabic, gelatine, talcum and titanium dioxide. Alternatively, the tablet may be coated with a suitable polymer dissolved in a readily volatile organic solvent.

For the preparation of soft gelatine capsules, the compounds of the invention may be admixed with, for example, a vegetable oil or polyethylene glycol. Hard gelatine capsules may contain granules of the compound using either the above-mentioned excipients for tablets. Also liquid or semisolid formulations of the compound of the invention may be filled into hard gelatine capsules. Liquid preparations for oral application may be in the form of syrups or suspensions, for example, solutions containing the compound of the invention, the balance being sugar and a mixture of ethanol, water, glycerol and propylene glycol. Optionally such liquid preparations may contain colouring agents, flavouring agents, sweetening agents (such as saccharine), preservative agents and/or carboxymethylcellulose as a thickening agent or other excipients known to those skilled in art.

For intravenous (parenteral) administration the compounds of the invention may be administered as a sterile aqueous or oily solution.

The size of the dose for therapeutic purposes of compounds of the invention will naturally vary according to the nature and severity of the conditions, the age and sex of the animal or patient and the route of administration, according to well known principles of medicine

Dosage levels, dose frequency, and treatment durations of compounds of the invention are expected to differ depending on the formulation and clinical indication, age, and co-morbid medical conditions of the patient. The standard duration of treatment with compounds of the invention is expected to vary between one and seven days for most clinical indications. It may be necessary to extend the duration of treatment beyond seven days in instances of recurrent infections or infections associated with tissues or implanted materials to which there is poor blood supply including bones/joints, respiratory tract, endocardium, and dental tissues.

Examples of isotopes suitable for inclusion in the compounds of the invention include isotopes

2 3 1 1 13 14 36 of hydrogen, such as H and H, carbon, such as C, C and C, chlorine, such as CI, fluorine, such as ¹⁸F, iodine, such as ¹²³l and ¹²⁵l, nitrogen, such as ¹³N and ¹⁵N, oxygen, such as ¹⁵0, ¹⁷0 and ¹⁸0, phosphorus, such as ³²P, and sulphur, such as ³⁵S.

Certain isotopically-labelled compounds, for example, those incorporating a radioactive isotope, are useful in drug and/or substrate tissue distribution studies. The radioactive isotopes tritium, i.e. ³H, and carbon-14, i.e. ¹⁴C, are particularly useful for this purpose in view of their ease of incorporation and ready means of detection.

Substitution with heavier isotopes such as deuterium, i.e. ²H, may afford certain therapeutic advantages resulting from greater metabolic stability, for example, increased in vivo half-life or reduced dosage requirements, and hence may be preferred in some circumstances.

Substitution with positron emitting isotopes, such as ¹¹C, ¹⁸F, ¹⁵0 and ¹³N, can be useful in Positron Emission Topography (PET) studies for examining substrate receptor occupancy. Isotopically-labelled compounds can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described using an appropriate isotopically-labelled reagent in place of the non-labelled reagent previously employed.

Throughout the description and claims of this specification, the words "comprise" and "contain" and variations of the words, for example "comprising" and "comprises", means "including but not limited to", and is not intended to (and does not) exclude other moieties, additives, components, integers or steps.

Throughout the description and claims of this specification, the singular encompasses the plural unless the context otherwise requires. In particular, where the indefinite article is used, the specification is to be understood as contemplating plurality as well as singularity, unless the context requires otherwise.

Features, integers, characteristics, compounds, chemical moieties or groups described in conjunction with a particular aspect, embodiment or example of the invention are to be understood to be applicable to any other aspect, embodiment or example described herein unless incompatible therewith.

Example 1 - Preparation of quinolone imidazole derivatives (Method A).

Method

To a solution of the starting material carbonyl diimidazole (0.2g) in dimethylformamide (5ml) under argon atmosphere at room temperature was added the required substrate (1 .1 equivalent; levofloxacin is shown in the scheme above). The reaction was stirred overnight then quenched with water (20ml), extracted using dichloromethane (x 3) and dried (MgS0₄). The products were purified using automated grace flash chromatography. In the case of levofloxacin the gradient elution was MeOH 10%/ DCM 90%.

Example 2 - Preparation of compounds of the invention (Method A)

To a solution of the starting material imidazole (0.2g; levofloxacin imidazole is shown as an example in the scheme above), in dimethylformamide (5ml) under argon atmosphere at room temperature was added the relevant hydrazine, hydrazide or hydroxylamine (1 .1 equivalent). The reaction was stirred overnight then quenched with water (20ml), extracted using dichloromethane (x 3) and dried (MgS0₄). The products were purified using automated grace flash chromatography. In the case of levofloxacin the gradient elution MeOH 10%/ DCM 90%.

Example 3 - Preparation of compounds of the invention (Method B)

To a solution of fluoroquinolone (e.g. levofloxacin hemihydrate, 200. mg, 0.5500mmol) in DCM (2ml_) was added triethylamine (0.08ml_, 0.5500mmol) followed by ethyl chloroformate

(0.05ml_, 0.5500mmol) at 0°C under Ar. This was allowed to stir at that temperature for 5 minutes. After which time the reaction mixture was allowed to warm to room temperature and hydrazine (0.5500mmol) was added in one portion. This was then allowed to stir at that temperature for 18 hours. After which time the reaction mixture was directly purified by automated flash chromatography (Grace Reveleris®) using an eluent system of DCM:MeOH.

Example 4 - Preparation of compounds of the invention (Method C)

To a room temperature solution of fluoroquinolone (1.25mmol) in MeCN (10ml_) was added trifluoroacetic anhydride at room temperature (1.21 ml_, 8.72mmol). The solution was left to stir for two hours and hydrazine (0.22ml_, 2.49mmol) was added, followed by triethylamine

(0.52ml_, 3.74mmol) approx. 20 minutes later. The solution was left to stir overnight, after which time the solution was diluted with DCM (10ml) and washed with water (4 x 10ml). The organic layer was dried (MgS0₄) and the solvent removed under vacuum. The resulting residue was purified by automated GRACE chromatography to give the desired product.

Exemplary compounds of the invention:

In an embodiment, the compound of the invention is any one or is selected from any group more than one of the following exemplary compounds:

B2013/050943



Unless indicated otherwise, the NMR data provided below was obtained in CDCI₃ using a 400 MHz NMR machine.

(A) δ 1.59-1.57 (3H, m), 2.37 (3H, s), 2.56-2.59 (4H, m), 3.32-3.43 (4H, m), 4.38-4.39 (1H, m), 4.41-4.48 (2H, m), 4.50 (2H, d), 5.33-5.41 (2H, m), 6.03-6.11 (1H, m), 7.67- 7.70 (1H, d), 8.60 (1H, s), 12.17 (1H, s). Compound A was prepared using method A.

(B) δ 1.53-1.54 (3H, d), 2.36 (3H, s), 2.54-2.58 (4H, m), 3.31-3.41 (4H, m), 4.27-4.36 (3H, m), 5.02-5.09 (2H, q), 7.27-7.61 (6H, m), 8.57 (1H, s), 12.14 (1H, s). Compound B was prepared using method A.

(C) δ 1.58-1.60 (3H, d), 2.37 (3H, s) 2.58 (4H, 4), 3.32-3.43 (4H, m), 4.29-4.33 (1H, q), 4.40-4.47 (2H, m), 5.17-5.30 (2H, d), 7.63-7.66 (1H, d), 8.56 (1H, s), 12.21 (1H, S). Compound C was prepared using method A.

(D) δ 1.51-1.52 (3H, d), 2.37 (3H, s), 2.56 (4H, m), 3.32-3.48 (4H, m), 4.18-4.21 (1H, q), 4.28-4.31 (1H, q), 4.48-4.53 (1H, m), 7.00-7.03(1 H, m), 7.12-7.14(2H, m), 7.26-7.31 (2H, m) 7.70-7.73 (2H, d) 8.77 (1 H, s), 12.68 (1 H, s). Compound D was prepared using method A. (E) δ 1.37 (9H, s), 1.58-1.59 (3H, m), 2.37 (3H, s), 2.56 (4H, m), 3.35-3.40 (4H, m), 4.31-4.43 (3H, m), 7.69-7.72 (1H, d), 8.63 (1H, s), 11.96 (1H, s). Compound E was prepared using method A.

(F) δ 0.95-0.99 (2H, m), 1.12-1.16 (2H, m), 1.31-1.34 (2H, m), 1.36-1.39 (1H, t), 1.41-

I.43 (1H, s), 3.22-3.25 (4H, m), 3.50 (1H, m), 3.60-3.67 (4H, m), 4.15-4.19 (2H, m), 7.29-7.31 (1H, d), 7.95-7.98 (1H, d), 8.70 (1H, s). Compound F was prepared using method A.

(G) δ 1.52-1.57 (3H, m), 2.37 (3H, s), 2.57 (4H, m), 3.34-3.42 (4H, m), 3.38-3.40 (2H, m), 4.47-4.52 (1H, m), 7.61-7.62 (1H, m), 7.84 (1H, s), 7.98 (1H, s), 8.09 (1H, m), 8.20 (1 H, s), 8.64-8.67 (1 H, d), 11.91 (1 H, s). Compound G was prepared using method A.

(H) δ 1.59-1.62 (3H, m), 2.37 (3H, s), 2.56 (4H, m), 3.33-3.42 (4H, m), 4.33-4.40 (3H, m), 5.11-5.13 (1H, d), 7.55-7.58 (1H, d), 8.51 (1H, s), 11.19-11.21 (1H, d). Compound H was prepared using method A.

(I) δ 1.34 (3H, m), 2.38 (3H, s), 2.59 (4H, m), 3.35-3.42 (4H, m), 4.23-4.30 (2H, m), 4.52-4.57 (1H, q), 6.71-6.77(2H, m), 7.46-7.71 (3H, m), 8.20-8.21 (1H, m), 8.88 (1H, s),

I I .63 (1 H, d). Compound I was prepared using method A.

(J) MS (m/z, rel. intensity) 554.3 (M⁺H⁺, 100). Compound J was prepared using method A.

(K) δ 1.48-1.76 (12H, m), 1.88-1.93 (5H, m), 2.17 (3H, s), 2.73-2.78 (4H, m), 2.88-2.89 (3H, m), 3.13-3.16 (5H, m), 3.41-3.45 (4H, m), 4.30-4.45 (3H, m), 7.59-7.69 (1H, dd), 8.70 (1H, s), 10.90 (1H, s). Compound K was prepared using method A. (L) δ 1.53-1.59 (3H, m), 2.36-2.37 (3H, s), 2.55-2.57 (4H, m), 2.88-2.95 (1H, S), 3.34- 3.41 (4H, m), 4.39-4.59 (3H, 7.34-7.41 (2H, m), 8.19 (1H, m), 8.52 (1H, m), 8.60-8.69 (1H, m) 9.14 (1H, s), 12.52 (1H, s). Compound Lwas prepared using method A.

(M) δ 1.54-1.56 (3H, d), 2.36 (3H, s), 2.55 (4H, m), 3.31-3.42 (4H, m), 4.29-4.40 (3H, m), 7.07-7.08 (1 H, d), 7.51 (2H, s), 8.51 (1 H, s), 11.84-11.85 (1 H, d). Compound M was prepared using method A.

(O) δ 1.53-1.55 (3H, d), 2.39 (3H, s), 2.55-2.60 (4H, m), 3.33-3.50 (4H, m), 4.26-4.44 (3H, m), 6.49-6.50 (1H, d), 6.82-6.87 (2H, m), 7.12-7.17 (2H, m), 7.66-7.70 (1H, d), 8.61 (1 H, s), 11.45-11.46 (1 H, d). Compound O was prepared using method A.

(P) δ 1.54-1.56 (3H, d), 2.38 (3H, s), 2.56-2.59 (4H, m), 3.36-3.47 (4H, m), 4.31-4.52 (3H, m), 6.88-6.91 (1H, d), 7.70-7.75 (2H, m), 8.11-8.14 (1H, d), 8.58-8.61 (1H, s), 11.67-11.68 (1 H, d). Compound P was prepared using method A. ill δ 11.5 (1H, m), 8.6 (1H, s), 7.7 (1H, d), 7.3 (1H, m), 7.2 (1H, m), 6.9 (1H, m), 6.8 (1H, m), 6.7 (1H, m), 4.5 (1H, m), 4.3 (2H, m), 3.3-3.5 (4H, m), 2.6 (4H, m), 2.4 (3H, m), 1.5 (3H, m). Compound T was prepared using method A.

(U) δ 12.8 (1H, m), 10.4 (1H, m), 8.7 (1H, m), 8.4 (1H, m), 8.2 (1H, m), 7.8 (1H, m), 6.7 (1H, m), 4.4-4.6 (2H, m), 4.3 (1H, m), 3.9 (3H, s), 3.4 (4H, m), 2.6 (4H, m), 2.3 (3H, m), 1.6 (3H, m). Compound U was prepared using method A.

(V) δ 14.8 (1H, m), 8.7 (1H, m), 7.9-8.0 (5H, m), 7.4 (1H, m), 7.2 (1H, m), 7.1 (1H, m), 3.8-4.0 (4H, m), 3.6 (1H, m), 3.4 (4H, m), 2.0 (2H, m), 1.4 (2H, m), 1.2 (2H, m). Compound V was prepared using method A. (W) δ 8.6 (1 H, s), 8.2 (1 H, s), 7.8-8.0 (2H, m), 7.4 (2H, m), 7.2 (1 H, m), 7.1 (1 H, m), 3.8 (4H, m), 3.5 (1 H, m), 3.3 (4H, m), 1.4 (2H, m), 1.2 (2H, m). Compound W was prepared using method A.

(X) δ 8.6 (1 H, m), 7.8 (4H, m), 7.1 (2H, m), 4.2 (3H, m), 3.2 (4H, m), 2.8 (4H, m), 2.4 (3H, s), 1.7 (3H, m). Compound X was prepared using method A.

(Y) δ 1 1.4 (1 H, s), 8.6 (1 H, s), 7.8 (1 H, d), 7.2 (2H, m), 6.8 (2H, m), 6.3 (1 H, m), 4.3 (2H, m), 4.2 (1 H, m), 3.2 (4H, m), 2.6 (4H, m), 2.2 (3H, s), 1.5 (3H, s). Compound Y was prepared using method A.

(AA) δ 1 1.5 (1 H, m), 8.6 (1 H, m), 7.7 (1 H, d), 6.9-7.2 (4H, m), 4.5 (3H, m), 3.5 (4H, m), 2.7 (4H, m), 2.5 (3H, s), 1.6 (3H, m). Compound AA was prepared using method A.

(AE) δ 1.5 (d, 3H), 2.4 (s, 3H), 2.5 (m, 4H), 3.4 (m, 4H) 4.0 (s, 2H) 4.4 (m, 3H), 7-7.6 (m, 5H), 8.6 (s, 1 H). Compound AE was prepared using method A.

(AG) δ 1 1.5 (s, 1 H), 8.96 (m, 1 H), 8.0 (s, 2NH), 7.75 (m, 1 H), 7.1 (m, 1 H), 4.8 (m, 1 H), 4.4 (m, 1 H), 3.4 (m, 4H), 3.3 (m, 1 H), 2.45 (m, 4H), 2.35 (s, 3H), 2.3 (s, 3H), 1.57-1.59 (3H, d).

Compound AG was prepared using method A.

(AH) δ 8.96 (m, 1 H), 8.0 (s, 2NH), 7.8 (m, 1 H), 7.75 (m, 1 H), 7.4 (m, 2H), 7.1 (m, 1 H), 4.8 (m, 1 H), 4.4 (m, 1 H), 3.4 (m, 4H), 3.3 (m, 1 H), 2.45 (m, 4H), 2.3 (s, 3H), 1.57-1.59 (3H, d). MS (m/z, rel. intensity) 495.5 (M⁺ H⁺, 97). Compound AH was prepared using method A.

(AI) MS (m/z, rel. intensity) 524.5 (M⁺ H⁺, 100). Compound Al was prepared using method A.

(AJ) MS (m/z, rel. intensity) 432.5 (M⁺ H⁺, 92). Compound AJ was prepared using method A. (AK) MS (m/z, rel. intensity) 493.5 (M⁺ H⁺, 90). Compound AK was prepared using method A.

(ΑΙ_) δ 8.96 (m, 1 H), 8.0 (s, 2NH), 7.95 (m, 2H), 7.75 (m, 1 H), 7.7 (m, 2H), 6.75 (m, 1 H), 5.35 (s, 1 H) 4.8 (m, 2H), 4.4 (m, 2H), 3.4 (m, 4H), 3.3 (m, 1 H), 2.45 (m, 4H), 2.3 (s, 3H), 1.57-1.59 (3H, d) MS (m/z, rel. intensity) 495.5 (M⁺ H⁺, 95). MS (m/z, rel. intensity) 495.5 (M⁺ H⁺, 95). Compound AL was prepared using method A.

(AM) δ 8.5 (s, 1 H), 7.7 (m, 2H), 7.3 (m, 2H), 4.5-4.3 (m, 3H), 3.4 (m, 4H), 2.5 (m, 4H), 2.3 (s, 3H), 1.4 (3H, d). MS (m/z, rel. intensity) 514 (M⁺ H⁺, 100). Compound AM was prepared using method A.

(AN) δ 8.96 (m, 1 H), 8.05-7.5 (m, 6H), 4.4 (m, 3H), 3.4 (m, 4H), 2.45 (m, 4H), 2.3 (s, 3H), 1.6 (3H, d). MS (m/z, rel. intensity) 480 (M⁺ H⁺, 100). Compound AN was prepared using method A.

(AP) δ 1.208-1.226 (m, 3H), 1.472-1.489 (m, 3H), 2.297 (s, 3H), 2.494 (s, 4H), 3.286- 3.334 (m, 4H), 3.671-3.724 (m, 2H), 4.159-4.195 (q, 2H), 4.253-4.381 (m, 2H), 6.1 -6.2 (s, 1 H), 7.626 (d, 1 H), 8.477 (s, 1 H), 1 1.229 (s, 1 H). Compound AP was prepared using method A.

(AQ) δ 1.443-1.460 (d, 3H), 2.27 (s, 3H), 2.366 (s, 3H), 2.531-2.561 (m, 4H), 3.31 1 -3.450 (m, 5H) 4.189-4.274 (m, 2H), 6.787-6.814 (m, 2H), 6.982-7.048 (m, 2H), 7.600-7.613 (m, 2H), 8.678 (s, 1 H), 1 1.475 (s, 1 H). Compound AQ was prepared using method A.

(AR) δ 1.550-1.566 (m, 3H), 2.176 (s, 3H), 2.582 (s,4H), 3.366-3.417 (m, 4H), 4.370-4.404 (m, 3H), 7.043-7.1 1 1 (m, 4H), 7.676-7.707 (m, 1 H), 7.91 (s, 1 H), 8.60 (s, 1 H), 1 1.55 (s, 1 H).

Compound AR was prepared using method A. (AS) δ 1.5-1.6 (m,3H), 2.35 (s, 3H), 2.5-2.6 (m, 4H), 3.3-3.5 (m, 4H), 3.7 (s, 1H), 4.2-4.6 (m, 3H), 6.8 (m, 1H), 6.9 (m,1H), 7.0 (m, 1H), 7.3-7.5 (d, 1H), 8.2 (s, 1H), 8.3-8.6 (m, 2H), 12.0 (s, 1H). Compound AS was prepared using method A.

(AT) δ 1.5-1.6 (m, 3H), 2.3 (s, 3H), 2.4-2.6 (m, 4H), 3.2-3.4 (m, 4H), 4.2-4.4 (m, 3H), 6.8 (m, 3H), 7.0 (m, 2H), 7.6-7.8 (m, 1H), 8.5 (s, 1H), 11.4-11.5 (s, 1H). MS (m/z, rel. intensity) 536 (M⁺ H⁺, 100). Compound AT was prepared using method A.

(AU) δ 1.5-1.6 (m,3H), 2.35 (s, 3H), 2.5-2.6 (m, 4H), 3.3-3.5 (m, 4H), 4.3-4.6 (m, 3H), 6.8 (d,2H), 7.3 (s, 1H), 7.4(d, 2H), 7.6 (m, 1H), 8.6 (s, 1H), 11.5 (s, 1H). Compound AU was prepared using method A.

(AV) δ 1.5-1.6 (m,3H), 2.35 (s, 3H), 2.5-2.6 (m, 4H), 3.3-3.5 (m, 4H), ), 4.2-4.6 (m, 3H), 6.5 (bs,1H), 6.7 (m, 1H), 6.8 (m, 1H), 6.9 (m, 1H), 7.1 (s, 1H), 7.5-7.7 (m, 1H), 8.7 (s, 1H), 11.47 (s, 1H). MS (m/z, rel. intensity) 488 (M⁺H⁺, 100). Compound AVwas prepared using method A.

(AW) δ 1.4 (d, 3H), 2.4 (s, 3H), 2.6 (m, 4H), 3.4 (m, 4H) 4.3 (m, 3H), 6.4-6.9 (m, 3H), 7.7 (m, 3H), 8.6 (s, 1H), 9.5 (s, 1H), 11.6 (s, 1H). Compound AW was prepared using method A.

(AX) MS (m/z, rel. intensity) 466.4 (M⁺H⁺, 100). UV254nm. Compound AX was prepared using method A.

(AY) δ 1.44-1.46 (d, 3H), 2.27 (s, 3H), 2.366 (s, 3H), 2.531-2.561 (m, 4H), 3.311-3.450 (m, 5H) 4.189-4.274 (m, 2H), 4.6 (1H, m) 6.9-7.2 (m, 2H), 6.982-7.048 (m, 3H), 7.8 (d, 1H)-7.613 (m, 2H), 8.1 (s, 1H) 8.9 (s, 1H), 11.6 (s, 1H). MS (m/z, rel. intensity) 520.3 (M⁺, 100). UV 254nm. Compound AY was prepared using method A.

(AZ) δ 0.8 (s, 2H), 1.0 (s, 3 H), 1.1-1.3 (m, 2H), 1.5-1.6 (m, 3H), 1.8-1.9 (m,3H), 2.2-2.4 (m, 1H), 3.1-3.3 (m, 2H), 3.35-3.5 (m,3H), 3.6 (s, 3H), 3.8-3.9 (m, 3H), 4.0-4.1 (m, 2H), 4.5-4.6 (m, 1 H), 5.1 (m, 1 H), 7.7-7.8 (d, 1 H), 8.3 (s, 1 H), 8.8 (s, 1 H), 15.0 (s, 1 H). Compound AZ was prepared using method C.

(BB) MS (m/z, rel. intensity) 531 (M⁺ H⁺, 100). Compound BB was prepared using method B.

(BD) δ 1 1.1 1 (s, 1 H), 8.58 (s, 1 H), 7.70 (d, 1 H), 5.08 (m, 1 H), 4.46-4.31 (m, 4 H), 3.45-3.25 (m, 6 H), 2.62 (t, 2 H), 2.58 (br t, 4 H), 2.39 (s, 3 H), 2.28 (s, 3 H) and 1.60 (d, 3 H).

Compound BD was prepared using method B.

(BE) δ 1 1.51 (s, 1 H), 8.82 (s, 1 H), 7.59 (d, 1 H), 7.07 (m, 1 H), 6.92 (d, 1 H), 6.80 (m, 1 H), 6.39 (m, 1 H), 4.35 (m, 1 H), 4.1 1 (m, 2 H), 3.35 (m, 4 H), 2.55 (br t, 4 H), 2.36 (s, 3 H), 2.28 (s,

3 H) and 1.37 (d, 3 H). Compound BE was prepared using method B.

(BF) δ 12.02 (s, 1 H), 8.85 (s, 1 H), 7.44 (m, 3 H), 7.02 (m, 1 H), 4.59 (m, 1 H), 4.29 (m, 2 H), 3.39 (m, 4 H), 2.59 (br t, 4 H), 2.40 (s, 3 H) and 1.38 (d, 3 H). Compound BF was prepared using method B.

(BG) δ 10.79 (s, 1 H), 8.76 (s, 1 H), 7.70 (d, 1 H), 4.43 (m, 2 H), 4.30 (m, 1 H), 3.39 (m, 4 H), 2.72 (2 x (3 H, s)), 2.57 (br t, 4 H), 2.38 (s, 3 H), 1.58 (d, 3 H). MS (m/z, rel. intensity) 404 (M⁺ H⁺, 50). Compound BG was prepared using method B.

(BH) δ 1 1.53 (s, 1 H), 8.61 (s, 1 H), 7.76 (d, 1 H), 6.90 (m, 3 H), 4.36 (m, 3 H), 3.41 (m, 4 H), 2.60 (br t, 4 H), 2.40 (s, 3 H), 2.30 (s, 3 H), 2.25 (s, 3 H) and 1.57 (d, 3 H). Compound BH was prepared using method A.

(BI) δ 1 1.1 1 (s, 1 H), 8.59 (s, 1 H), 7.70 (d, 1 H), 4.43-3.34 (m, 7 H), 3.05 (br t, 4 H), 2.57 (br t,

4 H), 2.38 (s, 3 H) and 1.61 (d, 3 H). Compound Bl was prepared using method A. (BJ) δ 1 1.75 (s, 1 H), 8.70 (s, 1 H), 8.22 (m, 1 H), 7.78 (d, 1 H), 7.48 (m, 1 H), 7.21 (m, 1 H), 6.86 (m, 1 H), 4.42 (m, 3 H), 3.51 (m, 4 H), 2.77 (m, 4 H), 2.53 (s, 3 H) and 1.61 (d, 3 H).

Compound BJ was prepared using method B.

(BK) δ 1 1.93 (s, 1 H), 9.07 (s, 1 H), 7.63-7.10 (m, 5 H), 4.52 (m, 1 H), 4.24 (m, 1 H), 4.00 (m, 1 H), 3.39 (br t, 4 H), 2.60 (br t, 4 H), 2.41 (s, 3 H) and 1.07 (d, 3 H). MS (m/z, rel. intensity) 509 (M⁺ H⁺, 60). Compound BK was prepared using method B.

(BM) δ 12.13 (s, 1 H), 8.64 (s, 1 H), 8.15 (d, 1 H), 7.70 (d, 2 H), 6.18 (d, 1 H), 4.48 (m, 1 H), 4.40 (m, 2 H), 3.94 (s, 3 H), 3.40 (m, 4 H), 2.58 (br t, 4 H), 2.39 (s, 3 H) and 1.57 (d, 3 H). MS (m/z, rel. intensity) 485 (M⁺ H⁺, 100). Compound BM was prepared using method B.

(BN) δ 1 1.7 (1 H, s), 8.7 (1 H, s), 8.3 (1 H, s), 8.1 (1 H, d), 7.7 (1 H, d), 7.2 (1 H, br s), 6.9 (1 H, s), 4.6-4.3 (3H, m), 3.5-3.3 (4H, m), 2.7 (4H, s), 2.5 (3H, s), 1.6 (3H, d). MS (m/z, rel. intensity) 529 (M H^", 95). Compound BN was prepared using method B.

(BO) δ 7.9 (1 H, s), 7.8 (1 H, s), 7.7 (1 H, d), 7.5 (2H, d), 6.7 (2H, d), 4.9-4.4 (3H, m), 3.6-3.4 (3H, s), 3.3 (4H, s), 2.9 (4H, s), 2.7 (3H, s), 1.5 (3H, d). MS (m/z, rel. intensity) 531 (M⁺ H⁺, 100). Compound BO was prepared using method B.

(BQ) δ 12.2 (1 H, s), 8.7 (1 H, s), 8.0 (1 H, d), 7.7 (1 H, d), 6.0 (1 H, s), 4.6-4.3 (3H, m), 3.8 (4H, d), 3.6 (4H, d), 3.4 (4H, d), 2.6 (4H, s), 2.4 (3H, s), 1.6 (3H, d). MS (m/z, rel. intensity) 538 (M⁺ H⁺, 10). Compound BQ was prepared using method B.

(BP) δ 1 1.5 (1 H, s), 8.8 (1 H, s), 7.6 (2H, d), 7.4 (2H, d), 6.9 (2H, d), 4.4-4.2 (3H, m), 3.5-3.2 (4H, m), 2.6 (4H, s), 2.4 (3H, s), 1.5 (3H, d). MS (m/z, rel. intensity) 520 (M⁺ H⁺, 100).

Compound BP was prepared using method B. (BR) δ 12.1 (1H, s), 9.8 (1H, s), 9.2 (1H, s), 8.8 (1H, s), 8.3 (1H, d), 7.7 (1H, d), 7.3 (1H, d), 4.6-4.2 (3H, m), 3.5-3.3 (4H, m), 2.7 (4H, s), 2.4 (3H, s) and 1.6 (3H, d). MS (m/z, rel.

intensity) 542 (M⁺H⁺, 100). Compound BR was prepared using method B.

(BS) δ 11.9 (1 H, s), 8.8 (1 H, s), 7.5 (1 H, d), 6.9 (1 H. s), 4.8 (1 H, s), 4.4 (2H, q), 3.4 (4H, s), 2.6 (4H, s), 2.4 (3H, s), 1.5 (3H, d). Compound BS was prepared using method B.

(BT) δ 11.8 (1H, s), 8.7 (1H, s), 7.8-7.6 (2H, m), 7.3 (1H, d), 7.1 (1H, s), 6.9 (1H, s), 4.5 (1H, d), 4.4-4.2 (2H, m), 3.5-3.3 (4H, m), 2.6 (4H, s), 2.4 (3H, s), 1.5 (3H, d). MS (m/z, rel. intensity) 521 (M⁺H⁺, 100). Compound BT was prepared using method B.

(BU) δ 12.1 (1H, s), 8.6 (1H, s), 8.1 (1H, s), 7.8 (1H, d), 7.5 (2H, d), 4.6 (1H, s), 4.5-4.3 (2H, m), 3.4 (4H, s), 2.5 (4H, s), 2.3 (3H, s), 1.5 (3H, d). MS (m/z, rel. intensity) 521 (M⁺ H⁺, 100). Compound BU was prepared using method B.

(BV) (NMR measured in D₆-DMSO and not CDCI₃) δ 10.8 (1H, s), 8.8 (1H, s), 8.4 (1H, s), 7.7 (1H, d), 6.6 (1H, brs), 4.9 (1H, d), 4.6 (1H, d), 4.4 (1H, d), 3.8 (4H, m) 3.3 (4H, m, H₂0), 2.8 (3H, s), 1.4 (3H, d). MS (m/z, rel. intensity) 488 (M⁺ H⁺, 97). Compound BVwas prepared using method B.

(BW) δ 11.2 (1H, s), 8.7 (1H, s), 7.8 (1H, d), 4.5 (2H, d), 4.4 (1H, d), 3.6-3.4 (4H, m), 2.6 (4H, s), 2.4 (3H, s), 1.6 (3H, s), 1.2 (9H, s). MS (m/z, rel. intensity) 432 (M⁺ H⁺, 20). Compound BWwas prepared using method B.

(BX) δ 11.3 (1H, s), 8.8 (1H, s), 8.7 (1H, s), 7.8 (2H, d), 7.6 (1H, d), 6.7 (2H, d), 4.9 (1H, d), 4.5 (1H, d), 4.4 (1H, d), 3.2 (2H, s), 2.5-2.3 (7H, m), 2.2 (3H, s), 1.4 (3H, d). MS (m/z, rel. intensity) 496 (M⁺H⁺, 100). Compound BX was prepared using method B.

(BY) δ 11.2 (1 H, s), 8.6 (1 H, s), 7.8 (1 H, s), 7.2 (2H, d), 6.9 (2H, d), 6.6 (2H, br s), 6.3 (1 H, s), 4.8 (1H, brs), 4.2-2.8 (6H, m), 1.5 (10H, m). Compound BY was prepared using method B. (BZ) δ 1.53-1.57 (3H, t), 2.26 (3H, s), 2.42 (3H, s), 2.67-2.69 (4H, t), 3.32-3.35 (4H, t), 4.25- 4.30 (2H, q), 6.40-6.41 (1H, d), 6.80-6.82 (1H, d), 6.86-6.89 (2H, d), 7.03-7.05 (2H, d), 8.09- 8.12 (1H, d), 8.67 (1H, s), 11.52-11.53 (1H, d). MS (m/z, rel. intensity) 438 (M⁺ H⁺, 100).

Compound BZ was prepared using method B.

(CA) δ 1.41-1.45 (1H, t), 1.57-1.61 (3H, t), 2.40 (3H, s), 2.56-2.62 (4H, m), 2.76 (2H, s), 3.34- 3.44 (4H, m), 4.31-4.38 (2H, m), 4.38-4.45 (2H, m), 7.68-7.72 (1H, m), 8.59 (1H, s), 11.07 (1H, s). MS (m/z, rel. intensity) 390 (M⁺H⁺, 35). Compound CAwas prepared using method B.

(CB) δ: 4.2 (m, 2H), 4.5 (m, 1H, 4.7 (m, 2H), 6.8 (m, 3H), 7.2 (d, 2H), 7.8 (m, 2H), 8.2 (s, 1H), 8.6 (s, 1 H), 11.0 (s, 1 H). Compound CB was prepared using method B.

(CC) (NMR measured in CD₃OD and not CDCI₃) δ 1.55 (m, 3H), 2.1 (s, 3H), 2.72 (m, 4H), 3.33 (m, 4H), 3.4 (m, 1H), 4.5-4.6 (m, 3H), 4.87 (s, H₂0), 6.85 (d, 2H), 7.19 (d, 2H), 8.03 (s, 1H), 8.8 (s, 1H). Compound CC was prepared using method B.

(CD) (NMR measured in CD₃OD and not CDCI₃) δ 1.25 (9H, s), 1.52-1.53 (3H, d), 2.96 (3H, s), 3.18-3.24 (4H, m), 3.30-3.32 (3H, m), 3.35 (2H, s), 3.55-3.70 (6H, m), 4.40-4.46 (1H, d), 4.52- 4.55 (1H, d), 4.72-4.73 (1H, d), 6.80-6.82 (2H, d), 7.23-7.25 (2H, d), 7.55-7.58 (1H, d), 8.79 (1H, s). MS (m/z, rel. intensity) 508 (M⁺H⁺, 100). Compound CD was prepared using method B.

(CE) δ 1.57-1.59 (3H, d), 2.37 (3H , s), 2.56-2.59 (4H, m), 3.32-3.42 (4H, m), 4.33-4.42 (2H, m), 4.46-4.47 (1H, d), 6.81-6.84 (1H, m), 7.74-7.77 (1H, d), 7.76-7.78 (1H, d), 7.94-7.96 (1H, d), 8.43-8.44 (1H, d), 8.60 (1H, s), 12.56-12.58 (1H, d). MS (m/z, rel. intensity) 519 (MH^", 100). Compound CE was prepared using method B.

(CF) δ 11.5 (s, 1H), 8.75 (s, 1H), 7.70 (d, 1H), 7.1 (m, 1H), 6.72 (m, 3H), 6.5 (m, 1H), 4.4-4.2 (m, 3H), 3.35 (m, 4H), 2.55 (m, 4H), 2.35 (s, 3H), 2.3 (s, 3H), 1.45 (d, 3H). MS (m/z, rel.

intensity) 466 (M⁺H⁺, 100). Compound CF was prepared using method B. (CG) δ 1 1.5 (s, 1 H), 8.6 (s, 1 H), 8.0 (s, 1 H), 7.10-7.65 (m, 3H), 4.4-4.2 (m, 4H), 3.35 (m, 4H), 2.50 (m, 4H), 2.4 (s, 3H), 1.5 (d, 3H). Compound CG was prepared using method B.

(CH) δ 1 1.8 (s, 1 H), 8.55 (s, 1 H), 7.8 (d, 1 H) 6.95 (m, 3H), 4.4-4.1 (m, 3H), 3.4 (m, 4H), 2.50 (m, 4H), 2.4 (s, 3H), 1.5 (d, 3H). MS (m/z, rel. intensity) 488 (M⁺ H⁺, 95). Compound CH was prepared using method B.

(CI) δ 1 1.8 (s, 1 H), 9.25 (s, 1 H), 8.7 (s, 1 H), 8.15 (s, 1 H), 7.05-7.6 (m, 3H), 4.4-4.1 (m, 3H), 3.35 (m, 4H), 2.50 (m, 4H), 2.3 (s, 3H), 1.5 (d, 3H). MS (m/z, rel. intensity) 531 (M⁺ H⁺, 95). Compound CI was prepared using method B.

(CJ) MS (m/z, rel. intensity) 545.2 (M⁺ H⁺, 100). UV 254nm. Compound CJ was prepared using method B.

(CK) MS (m/z, rel. intensity) 496.35 (M⁺ H⁺, 100). UV 254nm. Compound CK was prepared using method B.

(CL) δ 1 1.6 (1 H, s), 8.8 (1 H, s), 8.0 (1 H, d), 7.2-6.5 (5H, m) 4.3 (1 H, m), 4.0 (2H, m), 3.3-2.5 (8H, m) 2.0-1.5 (6H, m), 1.3 (3H, d). MS (m/z, rel. intensity) 451.3 (M⁺ H⁺, 100). UV 254nm. Compound CL was prepared using method B.

Example 5

The in vitro efficacy of a series of compounds was assessed for activity against a range of bacterial strains. All test articles were stored in the dark at 4 °C following delivery. Immediately prior to use, approximately 1 mg of each compound was accurately weighed and dissolved in the appropriate volume of DMSO to give a stock concentration of 1.28g/L.

Strains Susceptibility tests were performed against a range of anaerobic bacterial strains: Details of the strains used are as follows.

Revival and Growth of the Strains

All strains were recovered from long-term storage at -80 °C by sub-culturing onto fresh blood agar plates and incubating at 35-37 °C in air, except for Streptococcus pneumoniae which was incubated in the presence of 5% C0₂, for 24 hours. Following visual checks to ensure purity and appropriate colony characteristics, isolates were deemed suitable for use.

Preparation of the Inoculum

The inocula for each bacterial strain were prepared by picking 5-10 distinct colonies from the culture plates and suspending them in 3ml of sterile saline. The inoculum was resuspended by vigorous shaking on a vortex mixer for 15s. The turbidity was then adjusted to McFarland standard 0.5 (1 -5 x 106 CFU/ml). The inoculum was further diluted in Mueller Hinton Broth for MIC tests to give a final inoculum in each well of 2-8 x 105 CFU/ml. For Streptococcus pneumoniae the Mueller Hinton broth was supplemented with 5% lysed horse blood (MHLB).

MIC Assay Conditions

MICs were tested in Mueller Hinton broth (MHLB for S. pneumoniae) in accordance with the appropriate CLSI guidelines. STEP 1: Addition of Test Article

a. A stock solution was prepared at a concentration of 1.28 g/L in DMSO. The stock was further diluted in Mueller Hinton broth (or MHLB) to give a top starting concentration of 128 mg/L in the assay. In addition, for each strain, a comparator control was included. The final concentration range for the comparator control (ciprofloxacin) was 0.03 -16 η^/Ι_.100μΙ_ of Mueller Hinton broth was dispensed into each well in columns 2-12. 200μΙ_ of the appropriate test compound solution (at 256mg/L) was dispensed into each well in column 1. b. 100μΙ_ aliquots were pipetted from column 1 wells and dispensed into column 2 with a multichannel pipette (± 2% coefficient of variation) thus diluting two-fold. 100 μΙ_ samples were then pipetted from column 2 wells and dispensed into column 3. The process was repeated through to column 10. The final 100 μΙ_ of diluted drug from column 10 was then discarded. Row 1 1 acted as a positive control (no drug or test article, organisms added), Row 12 acted as a negative control (no drug or test article, and no organisms added).

STEP 2: Addition of Bacterial Strains

100μΙ_ of the appropriate inoculum suspension in Mueller Hinton broth (or MHLB) was added to the appropriate wells. This resulted in a well containing 200μΙ_ final volume (made up of 100μΙ_ diluted compound or diluents and 100μΙ_ of inoculum or broth alone).

STEP 3: Incubation of Assay Plates

All plates were incubated in the dark at 35-37 °C in air for 18-24 hours. STEP 4: Reading of Plates

Plates were read visually and spectrophotometrically (450nm) where possible, 24 hours post inoculation. Endpoints of 50%, 80% and 100% inhibition were determined (or CLSI interpretation endpoints following visual examination). The results are presented in Table 1 (as MICsn along with levofloxacin) and further results are presented in Table 2 (as MICioo)- P201345WO specification as filed{804521 }[1 ].doc

Table 1 : MIC₅₀ for selected compounds and levofloxacin against non-resistant (susceptible) strains of Gram negative and Gram positive bacteria (indicated by a + or a - in parentheses after the species name)

Klebsiella Streptococcus

Pseudomonas Escherichia Acinetobacter pneumoniae Staphylococcus pneumoniae Staphylococcus Enterococcu

Compound aeruginosa {-) coli (-) baumannii (-) (-) aureus (+) w epidermidis (+) s faecal is (+)

E 128 8 32 128 64 128 128 128

C 128 32 64 128 128 128 128 128

A 128 16 32 128 32 128 64 128

N 8 0.25 1 16 4 16 2 16

H 2 0.25 0.25 4 0.5 2 1 4

G 2 0.25 0.25 2 0.5 2 0.5 4

I 16 0.25 1 16 4 2 2 16

D 128 32 64 128 128 64 128 128

B 128 64 64 128 128 128 128 128

K 128 16 32 128 128 32 128 128

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F 64 4 8 128 1 128 0.25 4

<0.25 <0.25 <0.25 <0.25 <0.25 <0.25 <0.25 <0.25

0

<0.25 <0.25 <0.25 <0.25 <0.25

P 0.5 0.5 0.5

M 64 32 32 128 16 2 16 16

Q 0.5 0.25 0.25 2 0.25 0.25 0.25 1

8 0.25 0.5 8 2 8 1 8

S 2 0.25 0.25 2 0.25 1 0.25 4

T 2 0.25 0.25 8 0.25 0.25 0.25 4

<0.25 <0.25 <0.25 <0.25 <0.25

levofloxacin 0.5 0.5 0.5

Table 2: MIC₁₀o for selected compounds against a variety of resistant strains and non-resistant (susceptible) strains of Gram positive and Gram negative bacteria.

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Example 6

The in vitro efficacy of a series of compounds was assessed for activity against a range of bacterial strains in either unbuffered Mueller-Hinton (MH) broth, or MH broth buffered to pH 6.2 or pH 8.3, and their activity compared with that of levofloxacin.

Method

The bacterial strains used during this study are detailed below

All strains were recovered from long-term storage at -80°C by sub-culturing onto fresh agar plates and incubating aerobically overnight at 37°C. Following visual checks to ensure purity and appropriate colony characteristics, isolates were deemed suitable for use.

Compounds were stored at 4°C in the dark. All samples were received in 100μΙ aliquots at a concentration of 1.28 mg/mL.

MICs were determined in MH broth in accordance with Clinical and Laboratory Standards Institute guidelines M07-A9 for aerobic bacteria, other than using flat-bottomed wells to allow spectrophotometric reading of MIC assays. MH broth was prepared according to the manufacturer's instructions and either left unmodified, or buffered as detailed below. All broths were then filter-sterilised through a 0.22 μηη filter before use. Unbuffered MH broth was found to have a pH of 7.18. • For MH broth pH 6.2, 3-(N-morpholino)propanesulfonic acidacid (MOPS) was added to a final concentration of 20 mM, and the pH adjusted with 1 M HCI.

• For MH broth pH 8.2, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) was added to a final concentration of 20 mM, and the pH adjusted with 1 M NaOH.

Five to 10 well-isolated colonies were picked and suspended in 3 mL sterile saline. The inoculum was suspended by vigorous shaking on a vortex mixer for 15 s. Turbidity was adjusted to McFarland standard 0.5 (1 -5 x 10⁸ CFU/mL). The inoculum was further diluted in the corresponding MH broth for MIC tests to give a final inoculum in each well of 2-8 x 10⁵ CFU/mL. a. Stock solutions of compounds were diluted in MH to give a maximum starting concentration of 64 μg/mL in the assay. 100 μΙ_ of MH was dispensed into each well in columns two to 12 of a 96 well plate. 200 μΙ_ of the appropriate test compound solution (128 μg/mL) was dispensed into each well in column one. b. Serial two-fold dilutions were performed from column one to column 10 to give a concentration range of 128 to 0.25 μg/mL of each compound in the assay. Columns 1 1 and 12 served as positive (no drug or test article, inoculum added), and negative (no drug, test article, or inoculum added) growth controls respectively.

For all strains, 100 μΙ of inoculum suspension was added to 100 μΙ_ of diluted compound to give a final volume of 200 μΙ_ and a final concentration range of 64 - 0.125 μg/mL.

Assay plates were incubated aerobically at 37°C for 18-20 h. Following incubation, plates were assessed both visually and spectrophotometrically at 490 nm.

Results

Table 3. MICs for 100, 80, and 50% inhibition of growth of E. coli ATCC 25922 at pH 6.2, 7.18 and 8.2. NG, No growth in positive control wells.

E. coli ATCC 25922 pH 6.2 (MOPS) Unbuffered (pH 7.18) pH 8.2 (HEPES)

Compound 100% 80% 50% 100% 80% 50% 100% 80% 50%

BX 4 4 2 0.25 0.25 <0.125 NG NG NG

AA <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 NG NG NG

0 0.5 0.5 0.25 <0.125 <0.125 <0.125 NG NG NG

Q 1 0.5 0.5 0.25 0.25 <0.125 NG NG NG

AV 0.25 0.25 0.25 <0.125 <0.125 <0.125 NG NG NG

AW 4 4 4 0.5 0.5 0.25 NG NG NG

CA 8 4 4 2 2 1 NG NG NG

CB <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 NG NG NG

Nadifloxacin <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 NG NG NG

Delafloxacin <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 NG NG NG

Levofloxacin <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 NG NG NG

Table 4. MICs for 100, 80, and 50% inhibition of growth of A. baumannii NCTC 13420 at pH 6.2, 7.18 and 8.2.

Table 5. MICs for 100, 80, and 50% inhibition of growth of S.aureus ATCC 29213 at pH 6.2, 7.18 and 8.2.

0 8 8 8 0.5 0.5 0.25 0.5 0.5 0.5

Q 4 4 2 2 2 2 4 4 4

AV 2 2 1 0.5 0.5 0.5 0.5 0.5 0.5

AW 8 8 8 4 4 4 2 1 1

CA 2 2 2 4 4 2 8 8 4

CB <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 <0.125

Nadifloxacin <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 <0.125

Delafloxacin <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 <0.125

Levofloxacin 0.25 0.25 <0.125 <0.125 <0.125 <0.125 0.25 0.25 <0.125

Table 6. MICs for 100, 80, and 50% inhibition of growth of S.aureus NRS74 at pH 6.2, 7.2 and 8.2.

Example 7

The data provided in Example 6 shows that there can be expected to be a considerable degree of activity variation in the activity at certain pHs for any given compound. Thus some compounds of the invention may be more active at low pH infection sites such as open wounds, whereas others may be more active at high pH infection sites such as urinary tract infections. The following study demonstrates that compound O, which was shown in Example 4 to have low activity at low pH, does as expected have very little activity in acidic wound sites.

Sprague-Dawley male rats 225-250g (at delivery) were used in these studies which were supplied by Charles River UK Ltd and were specific pathogen free. Following receipt at our facility rats were allowed to acclimatise for 5-7 days (weigh at the start of the experiment ~250- 275g).

Rats were housed in sterilized individual ventilated cages that expose the animals at all times to HEPA filtered sterile air. Rats had free access to food and water (sterile) and were provided with sterile aspen chip bedding (changed every 3-4 days). Additionally, during infection, rats had additional access to wet food if required to ensure they remained fully hydrated. The room temperature was 22°C +/- 1°C, with a relative humidity of 60% and maximum background noise of 56dB. Rats were exposed to 12 hour light/dark cycles.

Staphylococcus aureus (S. aureus) ATCC 29213 (MSSA), a well characterised strain, was used throughout the study.

Method

In this efficacy model, group sizes of 4 rats per treatment group (test articles, comparator or vehicle control) per time-point were used. An additional group of 3 rats was included to determine burden 1 hour post-infection as pre-treatment group (total 19 rats). Rats were euthanized 24 hours post treatment. All rats were rendered neutropenic by immunosuppression with cyclophosphamide at 150mg/kg 3 days before infection and 75mg/kg 1 day before infection by intraperitoneal injection. The immunosuppression regime leads to neutropenia starting 24 hours post the first administration which continues throughout the study. 24 hours post the second round of immunosuppression, rats were infected with S. aureus ATCC 29213 by intramuscular injection of 0.1 mL of bacteria into both lateral thigh muscles under temporary inhaled anaesthesia using ~6.54x10⁶ cfu/rat thigh. Rats were administered buprenorphine at ~0.03 mg/kg subcutaneously immediately post infection and 12 hours post infection for pain relief (buprenorphine causes delayed gastric emptying that can effect the PK following oral administration).

Table 7 shows the study treatment groups, treatment regimen and harvest (kill) time points. A total of 19 rats were used in the study (4 per time point plus 3 untreated controls).

Compound O was reconstituted freshly at each time point for immediate dosing in dimethyl sulfoxide (DMSO) giving a stock of 100mg/ml_, the DMSO stock was then diluted 1 :50 and 1 : 10 with water for injection (WFI) to provide dosing solutions at 2mg/ml_ and 10mg/ml_ respectively. Upon dilution solutions immediately turned cloudy, but after addition of 100- 200μΙ_ 1 M HCI the solutions became clear, non-particulate, straw coloured solutions (~pH3). Dosing solutions were delivered at 10ml_/kg orally in order to achieve 20mg/kg and 100mg/kg per dose. Vehicle solution (10% Dimethyl sulfoxide (DMSO), WFI, HCI), was prepared in the same manner minus the test article and delivered by oral administration at 10ml_/kg. Comparator Levofloxacin (Tavanic-5mg/ml_, Sanofi Aventis, UK) was also diluted in water for injection to 2mg/ml_ and delivered orally at 10ml_/kg in order to achieve 20mg/kg per dose. Following reconstitution, all test article solutions remained clear and non-particulate for the duration of the dosing period. Antibacterial treatment was initiated 1 hour post infection and delivered twice by oral gavage at 10 mL/kg (2.5ml_ per 250g rat) (see Table 1). All dosing solutions were well tolerated when administered via oral gavage at 10ml_/kg.

Table 7: Treatment groups, treatment regime, time of tissue harvest and number of rats per group.

Throughout the study rats had their clinical condition assessed. Animals were euthanized 24h after treatment. Following euthanasia rat weights were determined prior to both thighs being removed and weighed individually. Individual thigh tissue samples were homogenized in ice cold sterile phosphate buffered saline. Thigh homogenates were then quantitatively cultured on CLED agar, incubated at 37°C for up to 2 days and colonies counted. The data from the culture burdens were analyzed using the Kruskal-Wallis test, all pairwise comparisons (Conover-lnman), using Stats Direct v. 2.7.8. Culture burdens of the animals treated with test agents were compared to those of the vehicle control group. Note that each thigh has been treated as an independent sample for the statistical analysis.

Results

Table 8: Endpoint thigh tissue burdens (cfu/g; cfu = colony forming unit) for samples collected 24 hours post treatment

In conclusion, compound O failed to demonstrate any antibacterial efficacy in reducing Staphylococcus aureus ATCC 29213 (MSSA) burdens from the low pH localised sites of infection in this study. These results were entirely consistant with our understanding (see Example 6).

Example 8 - Biodefence

Several compounds of the invention were further tested for activity against strains which are known surrogates for bacterial strains associated with biowarfare. The testing methodology were the same as those described in Example 5. The activity is shown in Table 9 and the surrogate relationship is shown in Table 10.

Table 9: Activity of selected compounds against biodefense surrogate strains

Bacillus subtilis - QBR114769-04 positive 4

Burk olderia cepacia - QBR114769-09 negative 32

B Bacillus cereus - QBR114769-06 positive 1

Bacillus subtilis - QBR114769-05 positive 0.5000

Burkholderia cepacia - QBR114769-08 negative 8

Burkholderia cepacia - QBR114769-09 negative 4

Burkholderia cepacia - QBR114769-07 negative 16

Yersinia enterocolitica - QBR114769-03 negative 0.1200

Yersinia enterocolitica - QBR114769-02 negative 0.2500

Yersinia enterocolitica - QBR114769-01 negative 0.2500

Bacillus subtilis - QBR114769-04 positive 0.2500

P Bacillus subtilis - QBR114769-04 positive 1

Yersinia enterocolitica - QBR114769-01 negative 2

Burkholderia cepacia - QBR114769-09 negative 16

Yersinia enterocolitica - QBR114769-03 negative 1

Burkholderia cepacia - QBR114769-07 negative 32

Burkholderia cepacia - QBR114769-08 negative 32

Bacillus cereus - QBR114769-06 positive 4

Yersinia enterocolitica - QBR114769-02 negative 1

Bacillus subtilis - QBR114769-05 positive 2

Table 10: Surrogate relationships of biodefense strains

Preliminary studies show that compounds have comparable activity to the corresponding approved parent compounds.

This preliminary data indicates that the compounds have antibacterial activity and can be used as a medicament against one of the above bacterial strains.

Thus the compounds of the invention are active in the treatment of one or more of the above strains.

In some cases the activity is broad spectrum. In other cases the activity is selective over Gram-negative bacteria or Gram positive bacteria. In yet further cases the activity is selective over one or more strains.

In addition, the compounds are active against bacterial strains associated with biowarfare, particularly Gram positive strains.

Claims

Claims

1. A compound of formula (I):

X is C or N;

Y is O or NR³;

R¹ is independently selected from the group consisting: H , C_{1 -4} alkyl or Ac;

R² is independently selected from the group consisting: H , Ci-C₆ alkyl, C₂-C₆-alkenyl, C₂-C₆- alkynyl, C C₆ haloalkyi, C₃-C₈ cydoalkyi, C₃-C₈ heterocycloalkyl, -(CH₂)n-C₃-C8 cydoalkyi, - (CH₂)n-C3-C₈ heterocycloalkyl, aryl, -(CH₂)_n-aryl, -(CO)-aryl, -(CO)-(CH₂)_n-aryl, heteroaryl, -(CO)- heteroaryl, -(CH₂)_n-heteroaryl and -(CO)-(CH₂)_n-heteroaryl; wherein each n is independently 1 , 2, 3 or 4;

R³ is independently selected at each occurrence from: H , C_{1 -6} alkyl or Ac; or R² and R³, together with the nitrogen to which they are attached form a 3- to 7- membered ring which optionally contains an O, S or NR⁴ group;

R⁴ is independently at each occurrence H , C_{1 -6} alkyl or C^-C Ce-alkyl;

R⁵ is selected from the group consisting of: C C₄ alkyl, C C₄ haloalkyi, C₃-C₅ cydoalkyi, C₃-C₅ halocycloalkyl; unsubstituted phenyl; phenyl substituted with from 1 to 3 independently selected halogen atoms; unsubstituted pyridyl; and pyridyl substituted with from 1 to 3 independently substituents selected from the group consisting of: halo and NHR_a; wherein R_a is H or Ac;

R⁶ is selected from the group consisting of: H, C C₄ alkyl and C C₄ haloalkyl;

or alternatively, R⁵ and R⁶, together with the atoms to which they are attached to form a 4-6- membered ring which optionally contains an O or S atom; wherein the 4-6-membered ring is optionally substituted with 1 or 2 groups independently selected from halo and C C₄ alkyl;

R⁹ is selected from the group consisting: H, NHR^a or CrC₄-alkyl; wherein R_a is H or Ac;

R¹⁰ is independently selected from the group: H or F;

R¹¹ is selected from the group consisting of: an N-heterocycloalkyl group and a C₃-C₈ cycloalkyi group; wherein the N-heterocycloalkyl group comprises from 5 to 10 ring atoms and at least one nitrogen atom wherein the N-heterocycloalkyl group is optionally substituted with from 1 -3 groups independently selected from halo, tri(C₁-C₄ alkyl)silyloxy, hydroxyl, C C₄ alkyl, oxo or oxime.and wherein any nitrogen which does not attach the N-heterocycloalkyl group to the rest of the compound of Formula (I) is an NR_a group; and the C₃-C₈ cycloalkyi group is optionally substituted with at least one NHR_a group and optionally further substituted with from 1-3 groups independently selected from halo, hydroxyl, tri(C₁-C₄ alkyl)silyloxy, C C₄ alkyl, oxo or oxime; wherein R_a is H or Ac;

R¹² is absent or is selected from the group consisting of: H, OR¹⁶ and halo; wherein R¹⁶ is selected from the group consisting of: C C₄ alkyl and C C₄ haloalkyl; or R¹² and R⁵, together with the atoms to which they are attached form a saturated or unsaturated 5- to 7- membered ring which optionally contains an O, S or NR⁴ group; wherein the 5-7-membered ring is optionally substituted with 1 or 2 groups independently selected from halo, C C₄ alkyl and C C₄ haloalkyl; wherein if X is N, R¹² is absent; wherein each of the aforementioned alkyl, haloalkyl, cycloalkyi, halocycloalkyl, aryl (e.g.

phenyl) and heteroaryl (e.g. pyridyl) groups are optionally substituted, where chemically possible, by 1 to 3 substituents which are each independently selected at each occurrence from the group consisting of: oxo, imino, oximo, halo, nitro, cyano, hydroxyl, amino, N- heterocycloalkyl, S0₃R^b, S0₂R^b, S0₂NR^bR^bC0₂R^b C(0)R^b, CONR^bR^b, C C₄-alkyl, C C₄ haloalkyi, C C₄ alkoxy, and C C₄ haloalkoxy, wherein R^b is selected from H, C C^alkyl and C C₄-haloalkyl.

2. A compound of claim 1 , wherein the compound of formula (I) is a compound of formula (II):

A compound of claim 1 , wherein the compound of formula (I) is a compound of formuls

4. A compound of any one of claims 1 to 3, wherein R² is independently selected from the group consisting: H, C C₆ alkyl, C₂-C₆-alkenyl, C₂-C₆-alkynyl, C^Ce haloalkyi, C₃-C₈ cydoalkyi, C₃-C₈ heterocycloalkyl, -(CH₂)_n-C₃-C₈ cydoalkyi, -(CH₂)_n-C₃-C₈ heterocycloalkyl, aryl, -(CH₂)_n- aryl, heteroaryl, and -(CH₂)_n-heteroaryl; wherein each n is independently 1 , 2, 3 or 4.

5. A compound of any one of claims 1 to 3, wherein R² is aryl.

6. A compound of claim 5, wherein R² is a phenyl group substituted with from 1 to 5 groups independently selected at each occurrence from: halo, nitro, cyano, hydroxyl, amino, N- heterocycloalkyl, S0₃R^b, S0₂R^b, S0₂NR^bR^b C0₂R^b C(0)R^b, CONR^bR^b, C C₄-alkyl, C C₄ haloalkyl, C C₄ alkoxy, and C C₄ haloalkoxy.

7. A compound of claim 6, wherein R² is a phenyl group substituted with at least one electron-withdrawing group.

8. A compound of claim 6 or claim 7, wherein R² is a mono- para- substituted phenyl group.

9. A compound of any of claims 1 to 3, wherein R² is a heteroaryl group.

10. A compound of claim 9, wherein R² is a 6-membered heteroaryl group containing 1 -3- nitrogen atoms optionally substituted with from 1 to 4 groups independently selected at each occurrence from: halo, nitro, cyano, hydroxyl, amino, N-heterocycloalkyl, S0₃R^b, S0₂R^b, S0₂NR^bR^b C0₂R^b C(0)R^b, CONR^bR^b, C C₄-alkyl, C C₄ haloalkyl, C C₄ alkoxy, and C C₄ haloalkoxy.

1 1 . A compound of any of claims 1 to 3, wherein R² is a Ci-C₆ alkyl or Ci-C₆ haloalkyl group.

12. A compound of any of claims 1 to 1 1 , for therapeutic use.

13. A compound of claim 12, wherein the compound is for use in treating bacterial infections.

14. A compound of claim 13, wherein the compound is for use in treating a bacterial infection caused by one or more resistant strains of bacteria.

15. A compound of claim 13, wherein the compound is for use in treating an infection caused by a bacterial strain associated with biowarfare.