EA038487B1 - Thiazole derivatives, pharmaceutical composition comprising same and uses thereof - Google Patents

Abstract

The invention relates to compounds which are thiazole of Formula (I), or a pharmaceutically acceptable salt thereof. Furthermore, the invention relates to a pharmaceutical composition comprising said compounds, as well as the uses of the compounds and a composition for the treatment or prevention of bacterial infection and for the removal or reduction of antibiotic resistance in Gram-negative bacteria.

Description

Scope of invention

The present invention relates to compounds that are derivatives of thiazoles. The compounds of the invention find use in the prevention or treatment of bacterial infection. The invention also provides such compounds per se and pharmaceutical compositions containing such compounds. The compounds of the invention are useful as inhibitors of metallo-β-lactamase (MBL) enzymes. The compounds of the invention can be used in combination therapy, for example in combination with one or more antibiotics and possibly one or more inhibitors of serine β-lactamase (SBL) enzymes. This combination therapy finds particular applications for the prevention or treatment of bacterial infection caused by bacteria that are resistant to antibiotic treatment when used alone, especially when resistance is due to the presence of metal-β-lactamase and / or serine-β-lactamase enzymes. , and treatment with β-lactam antibiotics alone may be unsuccessful. In such cases, combination therapy may preserve the antibacterial effect of the β-lactam antibiotic.

Prior art

Bacteria, both in clinical and non-clinical settings, are becoming increasingly resistant to traditional antibiotics, and this resistance is becoming a serious clinical and epidemiological problem for human health. For example, it has been shown that single amino acid mutations in bacterial DNA-dependent RNA polymerase can reduce the binding affinity of this target enzyme to antibiotics, resulting in a high resistance index (FoR). One approach that takes into account FoR, which was considered earlier, is to develop a single agent that inhibits two related bacterial enzymes. Examples of such agents include hepotidacin, which inhibits two similar DNA-processing components of topoisomerase II and IV enzymes (GyrA and PerC), and zoliflodacin, which inhibits two similar ATP-hydrolyzing components of topoisomerase II and IV enzymes (GyrB and PerE). However, this approach is not always suitable in the case of other forms of resistance, for example, when microbial resistance to an antibiotic results from the production of a bacterial enzyme capable of inactivating the antibacterial drug.

In gram-negative bacteria, resistance to antibiotics (in particular, β-lactam antibiotics) often results from the production of β-lactamases by the microorganism. The β-lactamase enzymes include both metallo-β-lactamase (MBL) and serine-β-lactamase (SBL). In serine β-lactamase enzymes, a serine residue in the active center is used for hydrolysis of β-lactam rings by the mechanism of covalent catalysis, while in metallo-β-lactamase enzymes differing in structure, coordination of the metal ion Zn and hydroxide ion is used for hydrolysis of the β-lactam ring. Among the enzymes of bacterial β-lactamases, in particular in gram-negative bacteria and, more specifically, in enterobacteria, older serine-β-lactamase enzymes are supplemented by relatively recently discovered metallo-β-lactamases. Consequently, the resistance of gram-negative bacteria to β-lactam antibiotics is mainly due to the production of two types of β-lactamases by this type of microorganism.

As discussed above, antibiotic resistance in gram-negative bacteria often results from the production of β-lactamases by the microorganism, especially metallo-β-lactamases (MBL). MBL are critical resistance factors of increasing clinical relevance. In fact, due to their broad spectrum, strong carbapenemase activity and resistance to inhibitors, these enzymes can confer resistance to almost all β-lactam antibiotics.

MBLs were first discovered in the mid-1960s as being carried by mobile DNA elements only in species with low pathogenic potential. However, the 1990s saw a proliferation of genes encoding MBL among major gram-negative bacteria, leading to a public health crisis resulting from the worldwide proliferation of carbapenem-resistant enterobacteriaceae producing VIM-type metallo-β-lactamases and NDMs. -type (Veronaintegron-encoded metallo-β-lactamase and metallo-β-lactamase New Delhi).

The functional features of these enterobacteria include strong carbapenemase activity and resistance to clinically used β-lactamase inhibitors (clavulanate and sulfones). The activity against β-lactams differs among different metallo-β-lactamases, and the substrate specificity can range from a narrow range (for example, in the case of metallo-β-lactamase CphA from Aeromonas hydrophila) to an extended range (for example, in the case of metallo-β- VIM-type lactamases, which can catalyze the degradation of almost all classes of β-lactams, except for monobactams).

There are three main structural subclasses of MBL, which are characterized by significant internal diversity. Representatives of different subclasses differ not only in a high degree of sequence diversity, but also in the structure of their active centers. In enzymes of subclasses B1 and B3, the active site contains two zinc ions; the representatives of subclass B2 have an active

- The 1,038487 center contains only one zinc ion.

Acquired metal-β-lactamases have been found in strains of Enterobacteriaceae, Pseudomonas aeruginosa, Acinetobacter baumannii, and other gram-negative bacteria. Among the acquired MBLs, almost all enzymes belong to the B1 subclass, which indicates an overall higher propensity of the representatives of this subclass to capture and spread mobile genetic elements in comparison with the representatives of the B2 and B3 subclasses.

For example, subclass B1 contains IMP-type (metal-to-lactamase imipenemase), VIM-type and NDM-type enzymes.

IMP-type enzymes, including IMP-1, were first discovered in Japan in the late 1980s and have since been reported worldwide in enterobacteriaceae and gram-negative bacteria. IMP-type enzymes have broad substrate specificity and high affinity for cephalosporins and carbapenems, but they are characterized by low activity against temocillin.

VIM-type enzymes, including VIM-2, were first discovered in Europe in the late 1990s and have since been registered worldwide. VIM-type enzymes were originally found in P. aeruginosa and other gram-negative bacteria, but have since emerged in enterobacteria and have become a serious problem in some cases. More than 20 different VIM-allotypes are known, each of which is characterized by a certain geographic distribution, with the exception of VIM-1 and VIM-2, which are more widespread than the IMP-type enzymes. VIM-type metallo-β-lactamases exhibit an even broader substrate specificity than IMP-type enzymes, showing the ability to catalyze the hydrolysis of 6-a-methoxy-penicillins. In addition, VIM-type enzymes are unique among metallo-β-lactamases in that they have a high affinity for carbapenems.

New Delhi metal-in-lactamase 1 (NDM-1) is a novel metal-in-lactamase originally identified in a patient admitted to a hospital in New Delhi with Klebsiella pneumoniae infection. Subsequently, it was found that microorganisms of the Enterobacteriaceae family containing this new β-lactamase are widespread throughout India, Pakistan and Bangladesh and are now found in the UK and many other countries. New Delhi metal-β-lactamase 1 (NDM-1) is a 158 amino acid long polypeptide (accession number AB571289) capable of catalyzing the hydrolysis of a wide range of β-lactam antibiotics, including penicillins, cephalosporins and carbapenem antibiotics, which are the basis for treatment antibiotic-resistant bacterial infections.

Accordingly, there is an urgent need for new antibacterial compounds and compositions and methods of adjuvant therapy for the treatment of a bacterial infection, in particular a bacterial infection caused by bacteria expressing MBL enzymes. There is also an urgent need for new compositions for the treatment of bacterial infection caused by bacteria that exhibit high resistance, in particular when resistance to antibiotics (especially β-lactam antibiotics) arises from the production of one or more enzymes by bacteria capable of inactivating the antibacterial drug. means. The present invention addresses some or all of these problems.

Brief description of the essence of the invention

Previously, the present inventors reported in WO 2014/198849 that certain thiazole derivatives are inhibitors of metallo-β-lactamases, including NDM-1. We have now surprisingly found that the compounds of formula (I) are potent inhibitors of metal lactamases, including NDM-1, and have improved properties over the compounds described in WO 2014/198849. Thus, these compounds are useful in the treatment and prevention of bacterial infection, for example when used in combination with β-lactam antibiotics.

The inventors have also found that the compounds of formula (I) can preferably be used in combination with inhibitors of serine β-lactamase enzymes and other antibiotics such as β-lactam antibiotics, for example antibiotics of the carbapenem group. Such combination therapies are of particular importance in the prevention or treatment of bacterial infection caused by bacteria that exhibit a high degree of antibiotic resistance when used alone, especially when the bacterial infection is caused by bacteria that produce β-lactamase enzymes.

Accordingly, the present invention provides a compound that is a thiazole derivative according to formula (I) or a pharmaceutically acceptable salt thereof

- 2 038487

[FORMULA (1)] where

R ¹ represents H;

® represents a cyclic group selected from C6-C ₁₀ aryl group and a 5-10-membered heteroaryl group containing 1, 2 or 3 heteroatoms selected from O, S and N;

each R ^{2 is} independently selected from (1) halogen or R ⁸ , where each R ^{8 is} independently selected from CN and —C (O) NR ^f R ^g , where each of R ^f and R ^g is H; and (2) O (C _1-3 αalkyl), which may optionally be substituted with 3 halogen substituents;

m is 0, 1 or 2;

R ³ represents a hydrogen atom;

n is 0 or 1;

Z 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- and NR ¹⁰ C ( O) S-;

L is a bond or is C _1-3 αalkylene; or

L represents -C (R ¹⁰ ) = N-;

X is a bond or, if L is different from a bond or —C (R ¹⁰ ) = N—, then X is a bond or is selected from —NR ¹⁰ -, —O— and —C (NR ¹⁰ ) -;

p is 0 or 1;

A

i) R ⁴ is H and R ^{5 is} selected from H, —CN and C1-C3 alkyl, which is unsubstituted or substituted with NR ¹⁰ R ¹¹ ;

or R ⁴ together with R ⁵ forms, together with the atoms to which they are attached, a 5-6 membered heterocyclic group containing at least one saturated ring carbon atom and containing 1 or 2 heteroatoms selected from O, S and N; and ii) R ^{6 is} selected from H, —CN, and C ₁ -C 3 alkyl, which is unsubstituted or substituted by —NR10R ¹¹ ; and

R ⁷ , if present, is selected from H, —CN, and C1-C3 alkyl;

or R ⁶ together with R ⁷ , if present, forms, together with the atoms to which they are attached, a 5-6 membered heterocyclic group containing at least one saturated ring carbon atom and containing 1 or 2 heteroatoms selected from O, S and N; or

V

R ⁴ represents H;

R ⁵ together with R ⁶ forms, together with the atoms to which they are attached, a 5-6 membered heterocyclic group containing at least one saturated ring carbon atom and containing 1 or 2 heteroatoms selected from O, S and N; and

R ^7, if present, is selected from H, -CN and _C1-C3 alkyl;

R ¹⁰ and R ¹¹ are each H.

In addition, the invention relates to a specific compound which is 5 - [[3,5 difluoro-4- (guanidinocarbamoylamino) phenyl] sulfonylamino] thiazole-4-carboxylic acid or a pharmaceutically acceptable salt thereof.

In yet another aspect, the present invention relates to a pharmaceutical composition comprising a compound of the invention together with at least one pharmaceutically acceptable carrier or diluent.

In another aspect, the present invention relates to the use of compounds of the invention for the treatment or prevention of a bacterial infection.

In another aspect, the present invention relates to the use of compounds of the invention for eliminating or reducing antibiotic resistance in gram-negative bacteria.

In another aspect, the present invention relates to the use of a pharmaceutical composition of the invention for the treatment or prevention of a bacterial infection.

- 3 038487

Brief description of the figures

FIG. 1 shows the results of in vivo efficacy studies carried out using the compound of Example 2 described in this invention in a mouse model. The data demonstrate the inhibition of bacterial infection due to K. pneumoniae NTBC104 in mice with meropenem alone compared to the combination (meropenem + compound of example 2). Meropenem at a dose of 30 mg / kg reduced the bacterial load slightly, while the combination of meropenem at a dose of 30 mg / kg with the compound of example 2 at a dose of 30 mg / kg significantly reduced the bacterial load compared to meropenem alone, which demonstrates a decrease in the number of colony-forming units (CFU) up to 1.6 log ₁₀ .

FIG. 2 shows the cumulative improvement in the minimum inhibitory concentration (MIC) for meropenem using the compounds of Example 2 and Example 26 in a panel of clinical strains of Enterobacteriaceae expressing NDM-type enzymes (out of 196 isolates). The data demonstrate that when the compounds of either Example 2 or Example 26 are used at a concentration of 8 μg / ml, the effect of meropenem is enhanced to such an extent that nearly 90% of the strains exhibit an MIC of 8 μg / ml for meropenem. In contrast, meropenem alone at the same concentration is capable of arresting growth in only less than 1% of the strains, and within the parameters of this experiment, growth arrest in 90% of all strains could not be achieved with meropenem alone.

Detailed description of the invention

Definitions

_C1-C4 alkplnaya group represents a linear or branched alkyl group containing from 1 to 4 carbon atoms. The C ₁ -C ₄ alkyl group is often a C ₁ -C ₃ alkyl group or a C ₂ -C ₄ alkyl group. Examples of _C1-C4 alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl and tert-butyl.

_C1-C3 alkplanaya group usually represents a _C1-C2 alkyl group. _C1-C2 alkplnaya group is methyl or ethyl, suitably methyl. For the avoidance of doubt, if two alkyl groups are present, the alkyl groups may be the same or different.

A C ₂ -C ₄ alkenyl group is a linear or branched alkenyl group containing from 2 to 4 carbon atoms and having one or more, for example one or two, usually one, double bond. Typically, a 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. For the avoidance of doubt, if two alkenyl groups are present, the alkenyl groups may be the same or different.

A C ₂ -C ₄ alkynyl group is a linear or branched alkynyl group containing from 2 to 4 carbon atoms and having one or more, for example one or two, usually one, triple bond. Typically, a C ₂ -C ₄ alkynyl group is a C ₂ -C ₃ alkynyl group. Examples of C ₂ -C ₄ alkynyl groups include ethynyl, propynyl, and butynyl. For the avoidance of doubt, if two alkynyl groups are present, then the alkynyl groups may be the same or different.

A C ₁ -C ₄ alkylene group is an unsubstituted or substituted bidentate moiety resulting from the removal of two hydrogen atoms from a C 1 -C ₄ alkane. These two hydrogen atoms can be removed from the same carbon or from different carbon atoms. Typically, the C _{1 -C 4} alkylene group is a C 1 -C ₃ alkylene group. Examples of C 1 -C ₄ alkylene groups include methylene, ethylene, n-propylene, isopropylene, n-butylene, sec-butylene and tert-butylene. _C1-C4 alkylene group is typically a C ₁ -C ₂ alkylene group. The C ₁ -C ₂ alkylene group is methylene or ethylene, usually methylene. For the avoidance of doubt, if two alkylene groups are present, the alkylene groups may be the same or different.

A C ₂ -C ₄ alkenylene group is an unsubstituted or substituted bidentate moiety resulting from the removal of two hydrogen atoms from a C ₂ -C ₄ alkene. These two hydrogen atoms can be removed from the same carbon or from different carbon atoms. Typically, a C ₂ -C ₄ alkenylene group is a C ₂ -C ₃ alkenylene group. Examples of C ₂ -C ₄ alkenylene groups include ethenylene, n-propenylene, isopropenylene, n-butenylene, sec-butenylene, and tert-butenylene. The C ₂ C ₃ alkenylene group is usually C ₂ alkenylene, i.e. ethenylene. For the avoidance of doubt, if two alkenylene groups are present, the alkenylene groups may be the same or different.

A C ₂ -C ₄ alkynylene group is an unsubstituted or substituted bidentate moiety resulting from the removal of two hydrogen atoms from a C ₂ -C ₄ alkyne. These two hydrogen atoms can be removed from the same carbon or from different carbon atoms. Typically, a C ₂ -C ₄ alkynylene group is a C ₂ -C ₃ alkynylene group. Examples of C ₂ -C ₄ alkynylene groups include ethynylene, n-propynylene, isopropinylene, n-butynylene, sec-butynylene, and t-butynylene. The C ₂ C ₃ alkynylene group is typically C ₂ alkynylene, i.e. ethynylene. For the avoidance of doubt, if two alkynylene groups are present, then the alkenylene groups may be the same.

- 4 038487 or different.

The alkyl, alkenyl, alkynyl, alkylene, alkenylene or alkynylene group can be unsubstituted or substituted. Unless otherwise indicated, substituted alkyl, alkenyl or alkynyl groups usually contain one or more than one, for example 1, 2, 3 or 4, such as one, two or three, for example one or two, for example one substituent selected from halogen, —CN, —R ⁸ , —OR ¹⁰ and —NR ¹⁰ R ¹¹ , where R ¹⁰ and R ¹¹ are as defined herein. The substituents themselves on a substituted alkyl, alkenyl, or alkynyl group are generally unsubstituted, unless otherwise indicated. If more than one substituent is present, they may be the same or different.

Halogen is usually chlorine, fluorine, bromine or iodine, and is preferably chlorine, bromine or fluorine, especially chlorine or fluorine, especially fluorine.

A 3-10 membered carbocyclic group is a cyclic hydrocarbon containing 3 to 10 carbon atoms. The carbocyclic group can be saturated or partially unsaturated, but is usually saturated. A 3-10 membered partially unsaturated carbocyclic 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 carbocyclic group is usually a 4-10 membered carbocyclic group. Often, the 3-10 membered carbocyclic group is a 3-6 membered carbocyclic group, such as a 4-6 membered or 5-6 membered carbocyclic group. The 3-10 membered carbocyclic group can be a fused bicyclic group as defined herein. The 3-10 membered carbocyclic group may be a saturated 4-6 membered, preferably 5 or 6 membered, carbocyclic group. Examples of 3-6 membered saturated carbocyclic groups include cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl groups.

A 3-10 membered heterocyclic group is a cyclic group containing 3 to 10 atoms selected from C, O, N and S in a ring including at least one heteroatom and usually one or two heteroatoms. Typically, the heteroatom or heteroatoms are selected (s) from O, N and S, most typically from S and N, especially N. For example, if the heterocyclic group is indicated to be a nitrogen-containing heterocyclic group, then it contains one nitrogen atom and optionally an additional heteroatom selected from O, N and S. The heterocyclic group can be saturated or partially unsaturated. A 3-10 membered partially unsaturated heterocyclic group is a cyclic group containing 3 to 10 atoms selected from C, O, N and S in a ring and containing 1 or 2, for example 1, double bond.

The 3-10 membered heterocyclic group is usually a 4-10 membered heterocyclic group. Sometimes, the 3-10 membered heterocyclic group is a 3-6 membered heterocyclic group such as a monocyclic 4-6 membered heterocyclic group or a monocyclic 5-6 membered heterocyclic group. Alternatively, the 3-10 membered heterocyclic group may be a 9 or 10 membered fused bicyclic heterocyclic group (i.e., a fused heterobicyclic group).

Examples of 5- and 6-membered saturated heterocyclic groups include piperazine, piperidine, morpholine, 1,3-oxazinane, pyrrolidine, imidazolidine, and oxazolidine, including their quaternized derivatives as defined herein. Examples of 5- and 6-membered partially saturated heterocyclic groups include tetrahydropyrazine, tetrahydropyridine, dihydro-1,4-oxazine, tetrahydropyrimidine, dihydro-1,3-oxazine, dihydropyrrole, dihydroimidazole and dihydrooxazole, including their derivatives, which are quaternized this description.

Examples of 9- and 10-membered fused heterobicyclic groups include 9-membered fused heterobicyclic groups such as indoline, 2,3-dihydrobenzofuran, 2,3-dihydrobenzo [b] thiophene, 2,3-dihydro-1H-benzo [d] imidazole, 2,3-dihydrobenzo [d] oxazole, 2,3-dihydrobenzo [d] thiazole, benzo [d] [1,3] dioxole, 4,5,6,7-tetrahydrothiazolo [5,4-c] pyridine and 4,5 , 6,7tetrahydrothiazolo [4,5-c] pyridine, including their quaternized derivatives as defined herein; and 10-membered heterobicyclic groups such as 1,2,3,4-tetrahydroquinoline, 1,2,3,4-tetrahydroisoquinoline, chroman, isochroman, thiochroman, isothiochroman, 1,2,3,4 tetrahydroquinoxaline, 1,2,3 , 4-tetrahydroquinazoline, 1,4-dihydro-2H-benzo [d] [1,3] oxazine, 3,4-dihydro2H-benzo [b] [1,4] oxazine, 3,4-dihydro-2H-benzo [b] [1,4] thiazine, 1,4-dihydro-2H-benzo [d] [1,3] thiazine, 4Hbenzo [d] [1,3] dioxin and 2,3-dihydrobenzo [b] [1 , 4] dioxin, including their quaternized derivatives. Often, the fused heterobicyclic group contains 1, 2 or 3, preferably 1 or 2, nitrogen atoms.

For the avoidance of doubt, references to a heterocyclic group also include fused polycyclic ring systems, including, for example, fused bicyclic systems in which the heterocyclic group is fused to an aryl group. When the heterocyclic group is such a fused heterocyclic group, preferred examples are fused ring systems where a 5-6 membered heterocyclic group is fused with a phenyl group.

A C ₃ -C ₆ cycloalkylene group (also referred to as a C _3-6 cycloalkylene group) is

- 5,038487 is an unsubstituted or substituted bidentate moiety resulting from the removal of two hydrogen atoms from a saturated C ₃ -C ₆ carbocyclic group as defined herein. These two hydrogen atoms can be removed from the same carbon or from different carbon atoms. Examples of C _3-6 cycloalkylene groups include cyclopropylene, cyclobutylene, cyclopentylene, and cyclohexylene.

A C ₆ -C ₁₀ aryl group is a substituted or unsubstituted monocyclic or fused polycyclic aromatic group having 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. The preferred group is phenyl (benzene).

The 5-10 membered heteroaryl group is a substituted or unsubstituted monocyclic or fused polycyclic aromatic group containing from 5 to 10 atoms in the ring portion, including at least one heteroatom, for example 1, 2 or 3 heteroatoms, usually selected from O, S and N. Typically, the heteroaryl group is a 5 or 6 membered heteroaryl group, or a 9 or 10 membered heteroaryl group, preferably a 5 or 6 membered heteroaryl group. Preferably, the heteroaryl group contains 1, 2 or 3, preferably 1 or 2, nitrogen atoms.

Examples of 5- and 6-membered heteroaryl groups include pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyridine, pyridazine, pyrimidine, and pyrazine. Examples of 9- and 10-membered heteroaryl groups include 9-membered heteroaryl groups such as indole, benzothiophene, benzofuran, benzoxazole, benzothiazole, benzimidazole, imidazo [1,2-a] pyridine, [1,2,4] thiazolo [1 , 5-a] pyridine and imidazo [1,2a] pyrazine, including their quaternized derivatives; and 10-membered heteroaryl groups such as quinoline, isoquinoline, quinazoline, and quinoxaline.

For the avoidance of doubt, references to a heteroaryl group also include fused polycyclic ring systems, including, for example, fused bicyclic systems in which the heteroaryl group is fused to an aryl group. When the heteroaryl group is such a fused heteroaryl group, preferred examples are fused ring systems where a 5-6 membered heteroaryl group is fused with a phenyl group.

A fused bicyclic group is a group containing two cyclic moieties having a common bond between two atoms.

A carbocyclic, heterocyclic, aryl, or heteroaryl group may be unsubstituted or substituted as described herein. For example, a carbocyclic, heterocyclic, aryl 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; -CN; —OR ¹⁰ and —NR ¹⁰ R ¹¹ (where R ¹⁰ and R ¹¹ are as defined herein), unsubstituted C1C ₂ alkyl and R ² as depicted in formula (I) and defined herein. The substituents themselves on a substituted carbocyclic, heterocyclic, aryl, or heteroaryl group are usually unsubstituted, unless otherwise indicated.

The compounds of the invention may contain heterocyclic or heteroaryl groups containing at least one nitrogen atom. In such compounds, said nitrogen atom (s) are independently selected from secondary, tertiary, and quaternary nitrogen atom (s). A quaternary nitrogen atom is present when the compound contains a quaternized derivative of one or more monocyclic groups or fused bicyclic groups. As used herein, a quaternized derivative of a moiety such as a cyclic moiety is formed by attaching an additional alkyl group to a nitrogen in the moiety such that the valence of said nitrogen increases from 3 to 4 and the nitrogen becomes positively charged.

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 n-toluenesulfonic acid. Pharmaceutically acceptable bases include hydroxides of alkali metals (eg sodium or potassium), alkaline earth metals (eg calcium or magnesium), and organic bases such as alkylamines, aralkylamines, and heterocyclic amines. Hydrochloride salts and acetate salts, especially hydrochloride salts, are preferred.

In formula (I), the stereochemical configuration is given without limitation. In particular, compounds of formula (I) containing one or more chiral centers can be used in enantiomerically or diastereomerically pure form or in the form of a mixture of isomers. In addition, for the avoidance of doubt, the compounds of the invention can be used in any tautomeric form. Typically, the agent or substance described in this invention contains at least 50%,

6,038487 preferably at least 60, 75, 90 or 95% of a compound according to formula (I) which is enantiomerically or diastereomerically pure. Typically, the compound of the invention contains at least 60% by weight, such as at least 75, 90 or 95% of the individual enantiomer or diastereomer. Preferably, this compound is substantially optically pure.

For the avoidance of doubt, the terms thiazole derivative and thiazolyl-containing derivative may be used interchangeably and, unless otherwise indicated, refer to compounds of the invention such as compounds of formula (I).

Compounds of the invention

In formula (I):

R ¹ represents H;

represents a cyclic group selected from a C ₆ -C ₁₀ aryl group and a 5-10 membered heteroaryl group containing 1, 2 or 3 heteroatoms selected from O, S and N;

each R ^{2 is} independently selected from (1) halogen or R ⁸ , where each R ^{8 is} independently selected from CN and —C (O) NRfR ^g , where each of R ^f and R ^g is H; and (2) O (C _1-3 alkyl), which may optionally be substituted with 3 halogen substituents;

m is 0, 1 or 2;

R ³ represents a hydrogen atom;

n is 0 or 1;

Z is a bond or selected from -NR ¹⁰ C (O) -, -C (O) NR ¹⁰ -, -NR ¹⁰ C (O) NR ¹¹ -, -NR ¹⁰ C (O) O- and -NR ¹⁰ C (O) S-;

L is a bond or is C _1-3 alkylene; or

L represents -C (R ¹⁰ ) = N-;

X is a bond or, if L is different from a bond or —C (R ¹⁰ ) = N—, then X is a bond or is selected from —NR ¹⁰ -, —O— and —C (NR ¹⁰ ) -;

p is 0 or 1;

A

i) R ⁴ is H and R ^{5 is} selected from H, —CN, and C ₁ -C ₃ alkyl, which is unsubstituted or substituted with —NR ¹⁰ R ¹¹ ;

or R ⁴ together with R ⁵ forms, together with the atoms to which they are attached, a 5-6 membered heterocyclic group containing at least one saturated ring carbon and containing 1 or 2 heteroatoms selected from O, S and N; and ii) R ^{6 is} selected from H, —CN, and C ₁ -C ₃ alkyl, which is unsubstituted or substituted with —NR ¹⁰ R ¹¹ ; and

R ⁷ , if present, is selected from H, —CN, and C ₁ -C ₃ alkyl;

or R ⁶ together with R ⁷ , if present, forms, together with the atoms to which they are attached, a 5-6 membered heterocyclic group containing at least one saturated ring carbon atom and containing 1 or 2 heteroatoms selected from O, S and N; or

V

R ⁴ represents H;

R ⁵ together with R ⁶ forms, together with the atoms to which they are attached, a 5-6 membered heterocyclic group containing at least one saturated ring carbon atom and containing 1 or 2 heteroatoms selected from O, S and N; and

R ⁷ , if present, is selected from H, —CN, and C ₁ -C ₃ alkyl;

R ¹⁰ and R ¹¹ are each H.

In one particular embodiment of the invention, ® is a cyclic group selected from a phenyl group and a 5-6 membered heteroaryl group containing 1, 2 or 3 heteroatoms selected from O, S and N.

In one particular embodiment of the invention, ® is selected from phenyl, pyridazine, pyridine and thiazole.

In one particular embodiment of the invention, ® is phenyl.

In one particular embodiment of the invention, each R ^{2 is} independently selected from halogen, —CN, —C (O) NR ^f R ^g , where R ^f and R ^g are each H; -OCF3 and -OCH ₃ .

In one particular embodiment of the invention, each R ² is fluoro.

In one particular embodiment of the invention, n is 0.

In one particular embodiment of the invention, Z is a bond or is selected from —NR ¹⁰ C (O) -, —C (O) NR ¹⁰ - and —NR ¹⁰ C (O) NR ¹¹ -.

In one particular embodiment of the invention, Z is selected from —NR ¹⁰ C (O) -, —C (O) NR ¹⁰ - and —NR ¹⁰ C (O) NR ¹¹ -.

In one particular embodiment of the invention, L is a bond.

In one particular embodiment of the invention, L is C1-3 alkylene.

In one particular embodiment of the invention, X is a bond.

In one particular embodiment of the invention, p is 1 and R ⁷ is H or methyl.

- 7 038487

In one particular embodiment of the invention, R ⁴ is H.

In one particular embodiment of the invention, R ⁵ is H and R ⁶ is H or methyl.

In one of the private embodiments of the invention

R ¹ represents H;

® represents a phenyl group;

m is 2;

each R ^{2 is} independently halogen;

η is 0;

Ζ selected from -NR ¹⁰ C (O) -, -C (O) NR ¹⁰ - and -NR ^lu C (O) NR- _:

L is a bond or is Ci-alkylene;

X is a bond;

p is 1; and

R ⁴ represents H;

R ⁵ is selected from H, -CN and _C1-C2 alkyl;

R ⁶ is H or methyl, and R ⁷ is H or methyl.

In one of the private embodiments of the invention:

R ¹ represents H;

® is phenyl;

m is 2;

each R ² is fluorine;

η is 0;

Ζ represents -NR ¹⁰ C (O) NR ⁿ -;

L is a bond;

X is a bond;

p is 1 and R ⁷ is H;

R ⁴ represents H;

R ⁵ represents H; and

R ⁶ is H.

In one particular embodiment of the invention, R ⁴ , R ⁵ , R ⁶ and R ⁷ , if present, each represent a hydrogen atom.

For the avoidance of doubt, a heterocyclic group containing at least one saturated carbon atom in the ring contains a —CH ₂ - group within the ring, and one or both hydrogen atoms in the —CH ₂ - group may be substituted as defined herein. description. Typically, the saturated ring carbon is unsubstituted; those. a heterocyclic group containing at least one saturated carbon atom in the ring usually contains a —CH ₂ - group within the ring. Thus, a heterocyclic group containing at least one saturated ring carbon is saturated or partially saturated. A heterocyclic group containing at least one saturated ring carbon is not aromatic.

-8 038487

In particular, preferred compounds of the invention are those selected from • 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-fluoro-phenyl] - sulfonylamino] thiazole-4-carboxylic acid;

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

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

• 5 - [[4 - [(2-guanidinoacetyl) amino] -3-methoxyphenyl] sulfonylamino] thiazole-4-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;

- 9 038487 • 5 - [[4 - [[2- (4,5-dihydro-1H-imidazol-2-ylamino) acetyl] amino] -3,5-difluoro-phenyl] sulfonylamino] thiazole-4-carboxylic acid ;

• 5 - [[6 - [(2-guanidinoacetyl) amino] pyridazin-3-yl] sulfonylamino] thiazole-4-carboxylic 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-imino-propanoyl) amino] -3,5-difluorophenyl] sulfonylamino] thiazole-4-carboxylic acid;

• 5 - [[4 - [[3- (dimethylamino) -3-imino-propanoyl] amino] -3-fluoro-phenyl] - 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-

4-carboxylic acid;

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

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

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

• 5 - [[4 - [(3-amino-3-imino-2-methyl-propanoyl) amino] -3-fluoro-phenyl] - 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 - [[(2P) -2-guanidinopropanoyl] amino] phenyl] sulfonylamino] - thiazole-4-carboxylic acid;

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

- 10 038487 • 5 - [[4 - [(4-amino-4-imino-butanoyl) amino] -3-fluorophenyl] sulfonylamino] thiazole-4-carboxylic acid;

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

• 5 - [[2,5-difluoro-4 - [(2-guanidinoacetyl) 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-1 - yl) acetyl] amino] phenyl] sulfonylamino] thiazole-4-carboxylic acid;

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

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

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

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

• 5 - [[4 - [(3-amino-3-imino-propanoyl) 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-4-carboxylic acid; and • 5 - [[4 - [(2-guanidinoacetyl) amino] phenyl] sulfonylamino] thiazole-4-carboxylic acid;

and their pharmaceutically acceptable salts.

In addition, the invention is a compound which is 5 - [[3,5 difluoro-4- (guanidinocarbamoylamino) phenyl] sulfonylamino] thiazole-4-carboxylic acid or a pharmaceutically acceptable salt thereof.

In one particular embodiment, the compound is 5 - [[3,5-difluoro-4 (guanidinocarbamoylamino) phenyl] sulfonylamino] thiazole-4-carboxylic acid.

In one particular embodiment, the compound is 5 - [[3,5-difluoro-4 (guanidinocarbamoylamino) phenyl] sulfonylamino] thiazole-4-carboxylic acid sodium salt.

Synthesis

The compounds of the invention can be prepared by any suitable method. Detailed general routes for the synthesis of representative compounds of the invention are provided below and in the Examples section.

In general, usually the compounds of the invention can be obtained by reactions corresponding to the following scheme

- 11 038487

The starting material, SM, is readily available and can be obtained, for example, using the methods described in WO 2014/198849. The description of WO 2014/198849 regarding the formation of the SM compound and its analogs is incorporated by reference. As a result of the reaction of SM with a sulfonyl chloride-containing derivative A (reaction step 1), a thiazole-sulfonamide-containing derivative A (A) is obtained. Reacting A with WZLX-Pro (B) provides intermediate C. In the above Scheme, Q and W are reactive groups that react with each other to couple A with B to provide C. For example, Q can be bromine, a -Z'-W can be -C (O) NH ₂ , as a result of which Q and W interact with each other through chemical transformations according to Buchwald (this, in particular, corresponds to the case when n is 0 ). Alternatively, Q may be —NH ₂ , and —Z'-W may be —C (O) OH, whereby Q and W interact with each other in a standard peptide coupling reaction using reagents such as HATU. Other methods for coupling compounds are well known to those skilled in the art. In compounds B and C, the —NR ⁷ —Pro moiety is a protected amine moiety that can be deprotected to provide the amine by standard techniques such as acid catalyzed deprotection (to provide compounds D). Suitable protecting groups for the amine are well known to those skilled in the art and include Boc (tert-butoxycarbonyl) protecting groups. The amine can then be reacted to form a guanidine group as in compound E by reaction with known guanidinylating agents such as 1H-pyrazole-1-carboximidamide. In compounds of the invention where p is zero and thus an amidine group rather than a guanidine group is present, the above synthetic route can be modified so that Compound B contains a protected amidine group rather than the protected amine NR ⁷ -Pro. Suitable protecting groups for amidine are well known to those skilled in the art and include Boc (tert-butoxycarbonyl) protecting groups. In these cases, the reaction of A and B gives compound C ', which is then deprotected to give the desired amidine-containing product E'. Detailed synthetic routes for representative compounds of the invention are set forth below.

Therapeutic efficacy

The compounds of the present invention are therapeutically useful and therefore suitable for use in medicine. The compounds of the invention can be used in the treatment of a human or animal body. For the avoidance of doubt, a compound of the invention may be administered in the form of a solvate.

Also provided is a pharmaceutical composition comprising a compound of the invention together with a pharmaceutically acceptable carrier or diluent.

In one particular embodiment, the composition may further comprise a β-lactam antibiotic.

In one particular embodiment, the β-lactam antibiotic is selected from carbapenems, penicillins, cephalosporins, and penems.

In one particular embodiment, the β-lactam antibiotic is meropenem.

In one particular embodiment, the composition may further comprise a serine β-lactamase inhibitor.

In one particular embodiment, the serine β-lactamase inhibitor is a compound of formula (II) or a pharmaceutically acceptable salt thereof

where

G is selected from —CN and —C (O) NR ^j R ^k ;

R ^{k is} selected from -W and -QW; where W is selected from 5-6 membered heterocyclyl, R ^j and -N (R ^j ) 2; and

Q is selected from -NR ^j C (O) -, -C (O) -NR ^j -, C1-3αalkylene, -O-C1-3alkylene, and -N (R ^j ) -C1-3αalkylene;

each R ^{j is} selected from H and unsubstituted C _1-3 alkyl, preferably H.

In one particular embodiment, the serine β-lactamase inhibitor is selected from WCK4234, avibactam, relbactam, zidebactam and nakubactam, or a pharmaceutically acceptable salt thereof, preferably WCK4234 or a pharmaceutically acceptable salt thereof.

Typically, the composition contains up to 85% by weight of the compound of the invention. More typically, it contains up to 50 wt% inclusive of the compound of the invention. Preferred pharmaceutical compositions are sterile and pyrogen-free. In addition, if pharmaceuticals

- 12,038487 If the compositions contain a compound of the invention that is optically active, the compound of the invention is usually a substantially pure optical isomer.

A composition of the invention may be in the form of a kit containing instructions allowing the kit to be used in the applications described herein, or details as to which subjects the application may be for.

As explained above, the compounds of the invention are useful in the treatment or prevention of bacterial infection. In particular, they are inhibitors of metallo-β-lactamase (MBL) enzymes and are therefore useful in eliminating or reducing antibiotic resistance of gram-negative bacteria.

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 additives in antibiotic therapy, eg, chemotherapy regimens. Alternatively, they can be used in combination with antibiotics to enhance the action of the antibiotics. The compounds of the invention may find particular use in the treatment or prevention of 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 metallo-β-lactamase and / or serine-β-lactamase enzymes. Treatment or prevention of such an infection with β-lactam antibiotics alone may not be successful.

When a joint composition is prepared, these two active agents may be presented as a pharmaceutical composition comprising (1) a compound of the invention as described in this invention, and (2) an additional antibacterial compound; and (3) a pharmaceutically acceptable carrier or diluent.

As indicated above, the antibiotic is a β-lactam antibiotic. More preferably, the antibiotic is a β-lactam antibiotic selected from carbapenems, penicillins, cephalosporins, and penems. Examples of antibiotics in the carbapenem group include imipenem, meropenem, ertapenem, doripenem, and biapenem. Examples of penicillins include amoxicillin, ampicillin, ticarcillin, piperacillin, and cloxacillin. Examples of cephalosporins include cefazolin, ceftriaxone, ceftazidime, and ceftobiprol. 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 a carbapenem antibiotic, more preferably imipenem or meropenem, most preferably meropenem.

As indicated above, the compositions of the invention may further comprise a serine β-lactamase (SBL) inhibitor, that is, they may contain (1) a compound of the invention; (2) a serine β-lactamase (SBL) inhibitor; and (3) an antibiotic. These compositions are referred to herein as triple combinations. The ternary combinations contain the above three active agents (1) to (3), but may also contain additional active agents if desired.

In a triple combination, each of a compound of the invention, an SBL inhibitor, and an antibiotic may be present in a single composition, or separate compositions may be prepared from them. Alternatively, two of these components can be presented in a single composition, and the remaining component can be presented separately. In other words, the composition can be prepared on the basis of a compound of the invention together with an SBL inhibitor and an antibiotic; or the composition can be prepared on the basis of a compound of the invention together with an SBL inhibitor, while the antibiotic is presented separately; or the composition can be prepared on the basis of a compound of the invention together with an antibiotic, while the SBL inhibitor is presented separately; or the composition can be formulated on the basis of an SBL inhibitor together with an antibiotic, while the compound of the invention is presented separately; or separate compositions can be prepared based on each of the compounds of the invention, SBL inhibitor and antibiotic. In the case of preparing separate compositions, the components of such a triple combination can be entered simultaneously or separately. They can be presented in kit form, possibly along with instructions for their administration.

If a joint composition can be prepared based on two or more active agents, then these two or more active agents can be presented in the form of a pharmaceutical composition containing (1) a compound of the invention described in this invention; (2) a pharmaceutically acceptable carrier or diluent; and either one or both of (3) an antibiotic and (4) a serine β-lactamase (SBL) inhibitor.

As indicated above, more preferably, in the pharmaceutical combination of the invention, the SBL inhibitor is selected from WCK4234, avibactam, relbactam, zidebactam and nakubactam, or pharmaceutically acceptable salts thereof. The structures of WCK4234, avibactam, relbactam, zidebactam and nakubactam are shown below. Such SBL inhibitors are commercially available and / or can be synthesized according to published protocols available to those skilled in the art.

- 13 038487

For example, WCK4234 and its synthesis are described in WO 2013/038330 and WO 2015/114595. Avibactam and its synthesis are described in Ball M. et al., Org. Process Res. Dev., 2016, 20 (10), pp. 1799-1805 and US 2012/323010. Relebactam and its synthesis are described in WO 2009/091856. Zidebactam and its synthesis are described in WO 2015/110885. Nakubactam and its synthesis are described in WO 2014/091268 and US 2016/272641.

nakubaktam

More preferably, in the pharmaceutical combination of the invention, the SBL inhibitor is WCK4234 or a pharmaceutically acceptable salt thereof. Even more preferably, the SBL inhibitor is WCK4234 or a sodium salt thereof. A process for the preparation of WCK4234 sodium salt is described in WO 2015/114595.

In the triple combination of the invention, the antibiotic is a β-lactam antibiotic. Preferably, the β-lactam antibiotic is selected from carbapenems, penicillins, cephalosporins and pemen, more preferably the β-lactam antibiotic is a cabapenem antibiotic, preferably imipenem or meropenem, most preferably meropenem.

Therefore, it is most preferred that the pharmaceutical combination of the invention comprises (1) a compound of the invention; (2) an SBL inhibitor selected from WCK4234, avibactam, relebactam, zidebactam, and nakubactam, and pharmaceutically acceptable salts thereof, preferably WCK4234 or a pharmaceutically acceptable salt thereof; and (3) an antibiotic of the cabapenem group, preferably meropenem.

The compounds of the invention are also useful in the treatment or prevention of bacterial infection.

Thus, the present invention also relates to the use of the compounds of the invention for the treatment or prevention of a bacterial infection.

In addition, the present invention also relates to the use of the compounds of the invention for the elimination or reduction of antibiotic resistance in gram-negative bacteria.

In addition, the present invention also relates to the use of a pharmaceutical composition according to the invention for the treatment or prevention of a bacterial infection.

In the above treatment, the subject 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 monkeys raised for the sale of animals such as horses, cattle, sheep or pigs, and pets such as dogs, cats, mice, rats, guinea pigs, ferrets, gerbils, or hamsters. A subject can be any animal that is susceptible to infection with a bacterium.

The compounds and compositions described in this invention are useful for treating bacterial infection that occurs after relapse after the end of antibiotic treatment. Thus, these compounds and compositions can be used to treat a patient who has previously received antibiotic treatment in the event of (the same episode) of a bacterial infection.

The bacterium causing the infection can be any bacteria that expresses the metal enzyme.

- 14 038487 lo-in-lactamase or its analogue. Typically, the bacterium that causes the infection expresses the MBL enzyme. Typically, the bacterium is a gram-negative bacterium. In particular, a bacterium can be a pathogenic bacterium. Typically, the bacterial infection to be treated using the compounds of the invention is resistant to conventional antibiotic treatment when the traditional antibiotic is used as monotherapy.

Gram-negative bacteria, the resistance of which to antibiotics can be eliminated using the compounds of general formula (I), are bacteria that produce metallo-β-lactamases, which can be metal-β-lactamases of subclasses B1, B2 or B3, for example, IMP-type enzymes (including IMP-1), VIM-type (including VIM-1 and VIM-2), and NDM-type (including NDM-1). Typically, gram-negative bacteria express NDM-type MBL enzymes, VIM-type MBL enzymes and / or IMP-type MBL enzymes; more typically, the bacteria express NDM-type MBL enzymes and / or VIM-type MBL enzymes; in the most typical case, the bacteria express NDM-type MBL enzymes. Gram-negative bacteria can express one or more of the following enzymes: ACT-TYPE, CMY-4, CTX-M-3, CTX-M-15, IMP-1, IMP-28, KPC-2, NDM-1, OXA-48 , OXA-181, SHV-OSBL, SHV-11, SHV-12, TEM-OSBL, TEM-1, VIM-1 and / or VIM-19.

The bacterial infection can be caused by bacteria from the families of Enterobacteriaceae, Pseudomonas and / or Moraxella, more typically the bacterial infection is caused by bacteria from the families of Enterobacteriaceae and / or Pseudomonas, and most typically the bacterial infection is caused by bacteria from the family of Enterobacteriaceae. Bacterial infection can be caused by Pseudomonas (for example, Pseudomonas aeruginosa, Pseudomonas oryzihabitans, or Pseudomonas plecoglossicida), Klebsiella, Escherichia, Acinetobacter, or Burkholderia. For example, a bacterial infection can be caused by Klebsiella pneumoniae, Escherichia coli, Pseudomonas aeruginosa, Burkholderia cepacia, or Acinetobacter baumannii. The bacterial infection can be caused by Escherichia coli, Klebsiella pneumoniae, or Klebsiella oxytoca. The bacterium can be a conditionally pathogenic microorganism.

The compounds and compositions of the invention are useful for preventing or treating infection with the following strains:

NTBC020 (E. coli strain expressing NDM-1, TEM-1 and CTX-M-15); NTBC035-2 (K. pneumoniae strain expressing NDM-1, CMY-4 and SHV-11); NTBC104-1 (K. pneumoniae strain expressing NDM-1 and SHV-11); NTBC123 (K. pneumoniae strain expressing NDM-1); NTBC062 (K. pneumoniae strain expressing IMP-1 and TEM-1); NTBC024 (K. pneumoniae strain expressing VIM-19, TEM-1 and CTX-M-3); NTBC042 (E. coli strain expressing VIM-1, TEM-1, CTXM-15, SHV-12); NTBC055 (E. coli strain expressing VIM-1); and NTBC039 (K. oxytoca strain expressing IMP-28).

The compounds and compositions of the invention may also be useful for preventing or treating an infection caused by the following strains. The triple combination is particularly useful for preventing or treating infection with the following strains:

NTBC019 (K. pneumoniae strain expressing NDM-1, CTX-M-15 and OXA-181); NTBC185 (K. pneumoniae strain expressing SHV-OSBL, TEM-OSBL, NDM-1 and OXA-48); NTBC186 (K. pneumoniae strain expressing ACT-TYPE, VIM-1 and OXA-48); NTBC187 (K. pneumoniae strain expressing SHV-OSBL, NDM-1 and OXA-48); and NTBC188 ((K. pneumoniae strain expressing NDM-1 and KPC-2).

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

The compound and composition of the invention can be used to treat patients with carbapenem-resistant enterobacteriaceae (CRE). CREs can be found in populations or in hospitals and other settings that are typically associated with long-term patients and patients undergoing significant medical interventions, such as those typically treated in intensive care units ( ICU).

The compound and composition of the invention can be administered to a subject to prevent the onset or recurrence of one or more 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 a bacterium. Such a subject is administered a prophylactically effective amount of an 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 and composition of the invention can be administered to a subject for the purpose of treating one or more symptoms of a bacterial infection. In this incarnation, the subject usually demonstrates the presence of

- 15,038487 symptoms. Such a subject is administered a therapeutically effective amount of an agent or composition. A therapeutically effective amount is an amount effective to reduce the intensity of one or more symptoms of the disorder.

The compound and composition of the invention can be administered in a variety of dosage forms. Thus, he / she can be administered orally, for example in the form of tablets, lozenges, troches, aqueous or oily suspensions, dispersible powders or granules. The formulated composition of the invention can also be administered parenterally, whether it is subcutaneous, intravenous, intramuscular, intrasternal, transdermal, or infusion techniques. The compound and composition can also be administered as a suppository. Preferably, the compound and composition can be administered by inhalation (aerosolized administration) or intravenous administration, most preferably by inhalation (aerosolized administration).

A compound and composition of the invention is typically formulated for administration together with a pharmaceutically acceptable carrier or diluent. For example, solid oral forms may contain, together with the active compound, diluents, for example lactose, dextrose, sucrose, cellulose, corn starch or potato starch; lubricants such as silicon dioxide, talc, stearic acid, magnesium or calcium stearate and / or polyethylene glycols; binders such as starches, arabic gums, gelatin, methylcellulose, carboxymethylcellulose or polyvinylpyrrolidone; disaggregating agents such as starch, alginic acid, alginates or sodium starch glycolate; gas-emitting mixtures; coloring agents; sweeteners; moisturizing agents such as lecithin, polysorbates, lauryl sulfates; and generally non-toxic and pharmacologically inactive substances used in pharmaceutical compositions. Such pharmaceutical preparations can be prepared in a known manner, for example using mixing, granulating, tabletting, sugar-coating or film-coating processes.

The compound and composition of the invention can be formulated for inhalation (aerosolized) administration as a solution or suspension. The compound and composition of the invention can be administered via a metered-dose inhaler (MDI) or nebulizer, such as an electronic or jet nebulizer. Alternatively, the compound and composition of the invention can be formulated for inhalation administration in the form of a powdered drug, such compositions can be administered from a dry powder inhaler (DPI). When prepared for inhalation administration, the compound and composition of the invention can be delivered in the form of particles having a mass average aerodynamic diameter (MMAD) of 1 to 100 μm, preferably 1 to 50 μm, more preferably 1 to 20 μm, such as from 3 to 10 microns, for example from 4 to 6 microns. When a compound and composition of the invention is delivered as a nebulized aerosol, reference to particle diameters indicates the MMAD for aerosol droplets. MMAD can be measured by any suitable method such as laser diffraction.

Liquid dispersions for oral administration can be syrups, emulsions, and suspensions. Syrups can 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, sodium alginate, pectin, methylcellulose, carboxymethyl cellulose or polyvinyl alcohol. Suspensions or solutions for intramuscular injection or inhalation may contain, together with the active compound, a pharmaceutically acceptable carrier, for example sterile water, olive oil, ethyl oleate, glycols, for example propylene glycol and, if desired, a suitable amount of lidocaine hydrochloride.

Solutions for inhalation, injection or infusion may contain, for example, sterile water as a carrier, or preferably they may be in the form of sterile aqueous isotonic saline solutions. Pharmaceutical compositions suitable for delivery by injection, for example, transdermally, can also be used.

The subject is administered a therapeutically or prophylactically effective amount of a compound of the invention. The dose can be determined according to various parameters, in particular according to the compound used; the age, weight and condition of the subject to be treated; route of administration; and the required treatment regimen. In addition, 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 body weight, according to the activity of the particular inhibitor, age, weight and condition. the subject to be treated, the type and severity of the disease, and the frequency and route of administration. Preferred daily dosage levels are between 5 mg and 2 g.

If a compound of the invention is administered to a subject in combination with another active agent (for example, in the form of a pharmaceutical combination containing an antibiotic and possibly an SBL inhibitor), then the dose of the other active agent (for example, an SBL inhibitor and / or antibiotic) can be determined as follows: as described above. The dose can be determined according to various parameters, in particular according to the agent used; the age, weight and condition of the subject to be treated; the way

- 16,038487 introductions; and the required treatment regimen. In addition, 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 body weight, according to the activity of the particular inhibitor, age, weight and condition. the subject to be treated, the type and severity of the disease, and the frequency and route of administration. Preferred daily dosage levels are between 5 mg and 2 g.

The antibacterial properties of the compounds described in this invention means that they are also useful in eliminating bacterial infection in vitro, i. E. in addition to treating subjects such as humans or animals. Thus, according to the invention, there is also provided a cleansing composition containing a thiazole derivative of the formula (I) or a salt thereof. The cleaning composition may further comprise, for example, a detergent, a surfactant (including ionic and nonionic surfactants), a diluent, a bleach (including hypochlorite such as sodium hypochlorite or calcium hypochlorite, chlorine, chlorine dioxide, hydrogen peroxide or its adduct , sodium perborate and sodium percarbonate), alcohol (such as ethanol or isopropanol), or a disinfectant. Typically, the disinfectant can be selected from benzyl 4-chlorophenol, amylphenol, phenylphenol, glutaraldehyde, alkyldimethylbenzyl ammonium chloride, alkyldimethylethylbenzyl ammonium chloride, iodine, peracetic acid, and chlorine dioxide. Typically, the detergent can be an alkaline detergent such as sodium hydroxide, sodium metasilicate or sodium carbonate, or an acidic detergent such as hydrochloric acid, nitric acid, sulfuric acid, phosphoric acid, citric acid, or tartaric acid.

The following examples illustrate the invention. However, they do not limit the invention in any way. Therefore, it is important to understand that the specific 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 determinative.

Experiment Details General Synthesis Methodology

There are several similar synthetic routes for this class of compounds described by formula 1, which are described below, where R is understood to mean any substituent on the phenyl ring.

Scheme 1.

The preparation of the key thiazole-containing intermediate, tert-butyl 5 - {[(4methoxyphenyl) methyl] amino} -1,3-thiazole-4-carboxylate, has been previously described (WO 2014/198849) and is readily prepared to scale 100 g. Its interaction with a wide range of arylsulfonyl chlorides was carried out using catalysis with bases (such as pyridine, triethylamine or sodium hydride) to give sulfonamide-containing intermediates such as [A]. Other variants of the thiazole-containing starting material (eg, where R1 is ethyl; R2 is H) are also readily available or even commercially available. Many of the compounds described in this invention are obtained by carrying out the standard Buchwald reaction for bromophenylsulfonamide [A], for example, with protected glycinamides such as [B]. General acid catalyzed deprotection liberates the primary amine [C], which can be converted to guanidine [D] if desired (Scheme 1) using a guanidinylating reagent such as 1H-pyrazole-1-carboxyimidamide.

Scheme 2

Alternatively, instead of forming an aryl-nitrogen bond in aryl bromide using Buchwald chemical transformations, some aniline-containing intermediates, such as [E], can be reacted with an N-protected amino acid glycine using standard peptide coupling reagents such as HATU (Scheme 2). Deprotection and then guanidinylation again give [C] and [D], respectively. Anilines [E] are prepared from the corresponding nitro compounds by standard reduction or from bromine-containing intermediates in the Buchwald reaction using ammonia (see, for example, Scheme 4).

- 17 038487

Scheme 3.

In certain circumstances, for example when substituents on the aryl ring are in particular electron-withdrawing, neither Buchwald amidation nor amide formation using protected glycine derivatives is successful. In these situations, it is necessary to react aniline with highly reactive chloroacetyl chloride to produce intermediate [F]. Then, as a result of substitution using sodium azide, azidoacetamide [G] is formed, which can be reduced with standard reducing agents, providing [C] and [D] in the usual way (Scheme 3).

Scheme 4.

To obtain some ureide-containing derivatives, it is necessary to carry out a special synthesis (Scheme 4). For example, in the Buchwald reaction, the corresponding aniline is obtained by reacting conventional bromoarylsulfonamide with ammonia as the nitrogen-containing component. Activation of this aniline with 4-nitrophenylchloroformate affords compound [H], which reacts with BOC-protected hydrazine [I] to provide coupling product [J], possibly by forming an isocyanate intermediate derived from [H]. As a result of treatment with a weak acid, the BOC group, which can be guanidinylated, is removed to obtain a protected guanidine functional group. Then, after total removal of the protective BOC groups, p-methoxybenzyl groups and tert-butyl ester groups, guanidine [K] is obtained.

Scheme 5.

To obtain some analogs, a special glyoxamide-containing intermediate is required

- 18 038487 compound [M], which is synthesized by reacting ordinary aniline with 0.5 eq. fumaryl chloride to give the symmetric bis-amide [L]. After ozonolysis, labile glyoxamide [M] is obtained, which can be reacted with a number of nucleophiles, including bisBOC-protected aminoguanidine, to obtain [N]. Then, after general removal of the protective groups in the usual way, the corresponding imine [O] is obtained (Scheme 5).

Abbreviations.

ACN acetonitrile, Ason acetic acid, Ag (OTf) silver triflate, AIBN azobisisobutyronitrile, Vos tert-butoxycarbonyl, Vos ₂ O di-tert-butyl dicarbonate, Sz ₂ COz cesium carbonate, CFU colony-forming unit, Cui copper iodide, DCM dichloromethane, DIPEA Ν, Ν-diisopropylethylamine, DMAP 4-dimethylaminopyridine, DMF dimethylformamide, DMS dimethyl sulfide, DMSO dimethyl sulfoxide, dppf 1,1'-bis (diphenylphosphino) ferrocene, EDC.HCI M- (3-dimethylaminopropyl) -N'-ethylcarbodiimide hydrochloride, EtOAc ethyl acetate, EtOH ethanol, Et ₃ N triethylamine, HATU 1- [bis (dimethylamino) methylene] -1 H-1,2,3-triazolo [4,5- B] pyridinium-3-oxide hexafluorophosphate, HCI hydrochloric acid, HOBt hydroxybenzotriazole, H ₂ SO ₄ sulphuric acid, IPA isopropyl alcohol, Km Michaelis constant, Mel methyl iodide, MeOH methanol, Nbs N-bromosuccinimide, Na ₂ CO ₃ sodium carbonate, Na ₂ SO ₄ sodium sulfate, Pd ₂ (dba) ₃ tris (dibenzylideneacetone) dipalladium (0), PdCI ₂ (PPh ₃ ) ₂ bis (triphenylphosphine) palladium (H) dichloride, PdCI ₂ (dppf) [1,1'-bis (diphenylphosphino) ferrocene] -dichloropalladium (P), PMB lara-methoxybenzyl, TEA triethylamine, TES triethylsilane, TMSOK potassium trimethylsilanolate, TFA trifluoroacetic acid, TMSOTf trimethylsilyl trifluoromethanesulfonate, TFA trifluoroacetic acid, THF tetrahydrofuran, T ₃ P propylphosphinic anhydride, KT room temperature.

- 19 038487

Structure of (RuPhos) -palladium (I) phenethylamine chloride (as an adduct with MTBE (methyl tert-butyl ether), 1: 1) used in the Buchwald coupling steps (first generation palladium catalyst complex based on RuPhos (RuPhos-Pd G1) ) is shown below

Examples of

Common Methods

Spectra Ή NMR (nuclear magnetic resonance) shown at 300 or 400 MHz in solutions in DMSO-d ₆ (δ in ppm ^-1) using chloroform as the reference standard (7.25 million ^-1). 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 specified, are given in hertz (Hz).

The term preparative high performance liquid chromatography (HPLC) (MDAP) purified refers to the purification of compounds using a Mass Directed Automated Purification (MDAP) system on an Agilent 1260 Infinity device with a preparative XSelect CHS C18 column eluting with 0.1% formic acid solution. acids in water / acetonitrile and detection using liquid chromatography combined with quadrupole mass spectrometry (Quadrupole LC / MS).

Example 1 tert-Butyl 5 - [(4-methoxyphenyl) methylamino] thiazole-4-carboxylate (key intermediate 1).

A suspension of potassium tert-butylate (874 mg; 7.79 mmol) in anhydrous tetrahydrofuran (10 ml) was vigorously stirred at room temperature. A solution of tert-butyl isocyanoacetate (1.0 g; 7.08 mmol) in anhydrous tetrahydrofuran (5 ml) was added dropwise thereto, and the mixture was stirred at room temperature for 10 minutes. To this mixture, a solution of 4-methoxybenzyl isothiocyanate (1.27 g; 7.08 mmol) in anhydrous tetrahydrofuran (5 ml) was added dropwise at room temperature. After 2 h, the solution was poured into saturated NaHCO ₃ solution and extracted with ethyl acetate. The organic layer was dried using Na ₂ SO ₄ , filtered and concentrated in vacuo to dryness. The residue was purified by silica gel chromatography (eluted with 0-50% ethyl acetate / cyclohexane) to give the title product as a pale yellow solid (852 mg).

¹ H NMR (CDCls) δ: 7.81 (1H, m), 7.73 (1H, br s), 7.31-7.23 (2H, m), 6.92-6.85 (2H, m), 4.35 (2H, d), 3.80 ( 3H, s), 1.61 (9H, s).

M / z 321 (M + H) +.

Example 2. 5 - [[3,5-Difluoro-4 - [(2-guanidinoacetyl) amino] phenyl] sulfonylamino] thiazole-4-carboxylic acid.

a) tert-Butyl 5 - [(4-bromo-3,5-difluoro-phenyl) sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4carboxylate.

- 20 038487

A solution of tert-butyl 5 - [(4-methoxyphenyl) methylamino] thiazole-4-carboxylate (1 g; 3.12 mmol; 1 eq.) In THF (15 ml) was added to a suspension of NaH in THF (10 ml) at 0 ° C in an argon atmosphere. After 30 min, a solution of 4-bromo-3,5-difluoro-benzenesulfonyl chloride (1.0 g; 3.43 mmol; 1.1 eq.) In THF (15 ml) was added at 0 ° C under argon. The resulting reaction mixture was stirred at RT for 1 h, quenched with ice-cold water (20 ml) and extracted with ethyl acetate (2x20 ml). The combined organic layer was dried over Na ₂ SO ₄ , filtered and concentrated. The crude material was purified by trituration with diethyl ether (2x5 ml) to give a pale yellow solid (800 mg; 44%).

M / z 577.0 (M + H) ⁺ .

b) tert-Butyl 5 - [[4 - [[2- (tert-butoxycarbonylamino) acetyl] amino] -3,5-difluoro-phenyl] sulfonyl [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate ...

A solution of tert-butyl 5 - [(4-bromo-3,5-difluoro-phenyl) sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole 4-carboxylate (100 mg; 0.173 mmol; 1 eq.) In 1, 4-dioxane (5 ml) was purged with argon for 15 min. Then, tert-butyl-N- (2-amino-2-oxo-ethyl) carbamate (45 mg; 0.26 mmol; 1.5 eq.), K ₃ PO ₄ (110 mg; 0.521 mmol; 3 eq.), Pd ₂ (dba) ₃ (16 mg; 0.17 mmol; 0.1 eq.) And Xantphos (30 mg; 0.052 mmol; 0.3 eq.). The resulting reaction mixture was heated to 85 ° C for 16 h in a closed vial. The reaction mixture was allowed to cool to room temperature, filtered through a pad of Celite and the pad was washed with EtOAc (2x5 mL). The organic layer was concentrated under reduced pressure. The resulting crude compound was dissolved in ethyl acetate (25 ml), washed with water (10 ml) and brine (10 ml). The organic layer was dried over Na ₂ SO ₄ , filtered and concentrated. The crude material was purified by flash chromatography (eluting with 55% ethyl acetate in petroleum ether) to give a pale yellow solid (60 mg; 51%).

M / z 669.5 (M + H) ⁺ .

c) 5 - [[4 - [(2-Aminoacetyl) amino] -3,5-difluoro-phenyl] sulfonylamino] thiazole-4-carboxylic acid, trifluoroacetate.

TFA (4 ml) was added to tert-butyl-5 - [[4 - [[2- (tert-butoxycarbonylamino) acetyl] amino] -3,5 difluoro-phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole -4-carboxylate (300 mg; 0.448 mmol; 1 eq.) At RT and stirred for 4 h. TFA was evaporated under reduced pressure. The resulting crude product was triturated with diethyl ether (2x5 ml) and dried under high vacuum to give a pale yellow solid (150 mg; 85%).

M / z 393.3 (M + H) +.

d) 5 - [[3,5-Difluoro-4 - [(2-guanidinoacetyl) amino] phenyl] sulfonylamino] thiazole-4-carboxylic acid.

- 21 038487

To a stirred solution of 5 - [[4 - [(2-aminoacetyl) amino] -3,5-difluoro-phenyl] sulfonylamino] thiazole-4-carboxylic acid in the form of trifluoroacetate (150 mg; 0.382 mmol; 1 eq.) In DMF (5 ml) pyrazole-1-carboxamidine hydrochloride (84 mg; 0.573 mmol; 1.5 eq.) And DIPEA (0.3 ml; 1.91 mmol; 5 eq.) Were added at RT. The resulting reaction mixture was stirred at RT for 16 h and concentrated under reduced pressure. Water (5 ml) was added to the residue, the precipitate was filtered off and washed with diethyl ether (2x5 ml). The crude product was purified by preparative HPLC to give the title compound as a white solid (47 mg; 28%).

¹ H NMR (400 MHz, DMSO-de) δ 13.20 (1H, s), 10.14 (1H, brs), 8.12 (1H, s), 7.55 (1H, brs), 7.43 (2H, d, J = 7, 2 Hz), 7.35-7.10 (3H, brs), 4.12 (2H, s).

M / z 434.9 (M + H) ⁺ .

LC-MS conditions.

Column: Acquity VEN C18 (50 mm x 2.1 mm; 1.7 μm).

Mobile phase: A: 0.05% formic acid (FA) solution in water; B: 0.05% formic acid solution in ACN.

Time (min) /% B: 0/3; 0.4 / 3; 2/98; 3.4 / 98; 3.5 / 3; 4/3.

Column temperature: 35 ° C, flow rate: 0.6 ml / min.

Preparative HPLC conditions.

Column: Symmetry C18 (300x19mm, 7μm).

Mobile phase: (A): 0.1% formic acid solution, (B): acetonitrile.

Flow rate: 19 ml / min.

Gradient (T (time) /% B): 0/5; 1/5; 8/20; 11/20; 11.02 / 99; 12/99; 12.1 / 5; 15/5.

Solubility: ACN + H ₂ O + THF + FA.

Example 3.

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

a) 5-Fluoro-6-hydroxy-pyridine-3-sulfonyl chloride.

3-Fluoropyridin-2-ol (2 g; 17.6 mmol) was added to chlorosulfonic acid (20 ml; 300.3 mmol) at 0 ° C. The reaction mixture was stirred at 160 ° C for 2 h, cooled to RT and slowly poured into ice-cold water (50 ml). The aqueous layer was extracted with EtOAc (3x50 ml). The organic layer was dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The crude compound was triturated with n-pentane (2x50 ml) to give an off-white solid (2.7 g; 72%).

M / z 212.11 (M + H) +.

b) 6-Chloro-5-fluoro-pyridine-3-sulfonyl chloride.

Thionyl chloride (5 ml; 68.9 mmol) was added to 5-fluoro-6-hydroxy-pyridine-3-sulfonyl chloride (1 g; 4.73 mmol) in toluene (25 ml) at 0 ° C. Then DMF (0.2 ml) was added slowly at 0 ° C. The reaction mixture was boiled under reflux for 3 h, cooled to RT, and concentrated under reduced pressure. The resulting crude material was co-distilled with toluene (2x25 ml) to give a pale yellow liquid which was used in the next step without further purification (0.9 g crude).

1H NMR (400 MHz, DMSO-d ₆ ) δ 8.83 (1H, m), 8.04 (1H, m).

c) tert-Butyl 5 - [(6-chloro-5-fluoro-3-pyridyl) sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate.

- 22 038487

A solution of tert-butyl 5 - [(4-methoxyphenyl) methylamino] thiazole-4-carboxylate (1.5 g; 4.68 mmol) in THF (25 ml) was added to a NaH suspension (1.12 g; 46.8 mmol) in THF (10 ml) at 0 ° C under argon. After 15 minutes, a solution of 6-chloro-5-fluoro-pyridine-3sulfonyl chloride (1.6 g; 7.0 mmol) in THF (15 ml) was added to the above reaction mixture at 0 ° C. under argon. The resulting reaction mixture was stirred at RT for 0.5 h, quenched with ice-cold water (20 ml) and extracted with ethyl acetate (3x20 ml). The combined organic layer was dried over Na ₂ SO ₄ , filtered and concentrated. The crude compound was purified by flash chromatography (eluting with 10-15% ethyl acetate in petroleum ether) to give a yellow oil (1.3 g; 54%).

M / z 514.27 (M + H) +.

d) tert-Butyl-5 - [[6 - [[2- (tert-butoxycarbonylamino) acetyl] amino] -5-fluoro-3-pyridyl] sulfonyl

[(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate.

Solution of tert-butyl 5 - [(6-chloro-5-fluoro-3-pyridyl) sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate (150 mg; 0.29 mmol) in 1, 4-dioxane (5 ml) was purged with argon for 20 min. Then tert-butyl-N- (2-amino-2-oxo-etul) carbamate (75 mg; 0.43 mmol), Cs ₂ CO ₃ (282 mg; 0.87 mmol), Pd ₂ (dba) ₃ (26 mg; 0.02 mmol) and Xantphos (50 mg; 0.08 mmol) in argon atmosphere. The resulting reaction mixture was heated to 70 ° C for 0.5 h in a sealed tube, allowed to cool to RT, filtered through a pad of Celite and the pad was washed with ethyl acetate (2x3 ml). The organic layer was concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with 50% ethyl acetate in petroleum ether) to give a pale yellow solid (75 mg; 39%).

M / z 652.41 (M + H) ⁺ .

f) 5 - [[6 - [(2-Aminoacetyl) amino] -5-fluoro-3-pyridyl] sulfonylamino] thiazole-4-carboxylic acid, trifluoroacetate.

N

911 about

S ^ NH o

TFA (1.5 ml) was added to a solution of tert-butyl-5 - [[6 - [[2- (tert-butoxycarbonylamino) acetyl] amino] -5-fluoro-3-pyridyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate (150 mg; 0.23 mmol) in DCM (2 ml) at 0 ° C, stirred at RT for 18 h and concentrated under reduced pressure. The residue was triturated with diethyl ether (2x2 ml), n-pentane (2x2 ml) and dried under high vacuum to give an off-white solid which was used in the next step without further purification (60 mg crude).

M / z 376.24 (M + H) ⁺ .

f) 5 - [[5-Fluoro-6 - [(2-guanidinoacetyl) amino] -3-pyridyl] sulfonylamino] thiazole-4-carboxylic acid.

Pyrazole-1-carboxamidine hydrochloride (70 mg; 0.48 mmol) and DIPEA (0.27 ml; 1.6 mmol) were added to a stirred solution of 5 - [[6 - [(2-aminoacetyl) amino] -5-fluoro-3-pyridyl] sulfonylamino] thiazole-4-carboxylic acid in the form of trifluoroacetate (120 mg; 0.32 mmol) in DMF (2 ml) at RT. The resulting reaction mixture was stirred at RT for 4 h, concentrated under reduced pressure, and ice-cold 1N hydrochloric acid was added to the crude compound. HCl solution (2 ml) and stirred for 10 min. The resulting precipitate was filtered off, washed with diethyl ether (2x5 ml) and dried under high vacuum. The crude product was purified by preparative HPLC to give the title product as an off-white solid (25 mg; 18%).

Ή NMR (400 MHz, DMSO-de) δ 13.20 (1H, brs), 10.8 (1H, brs), 8.51 (1H, d, J = 1.6Hz), 8.13 (1H, s), 7.99 (1H, dd, J = 9.6 Hz, J = 1.6 Hz), 7.52 (1H, brs), 7.26 (3H, brs), 4.20 (2H, d, J = 4.4 Hz).

M / z 418.18 (M + H) +.

LC-MS conditions.

Column: Acquity VEN C18 (50 mm x 2.1 mm; 1.7 μm).

Mobile phase: A: 0.05% formic acid solution in water; B: 0.05% formic acid solution in ACN.

Time (min) /% B: 0/3; 0.4 / 3; 3.2 / 98; 3.8 / 98; 4.2 / 3; 4.5 / 3.

Column temperature: 35 ° C.

Flow rate: 0.6 ml / min.

Preparative HPLC conditions.

Column used: PHENYL HEXYL (150x30 mm, 5 μm).

Mobile phase: (A): 0.1% formic acid solution, (B): acetonitrile.

Flow rate: 19 ml / min.

Gradient (T /% B): 0/5; 1/5; 6/30; 8.9 / 30; 8.95 / 99; 11/99; 11.1 / 5; 14/5.

Solubility: ACN + THF.

Example 4.

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

n θΜΑ ° ^NH2

S _NH

a) tert-Butyl N- [3-amino-1- (tert-butoxycarbonylamino) -3-oxo-prop-1-enyl] carbamate.

About NHBoc

Saturated NaHCO ₃ solution (10 ml) was added to a stirred solution of 3-amino-3-iminopropanamide (3 g; 29.6 mmol) in dioxane (20 ml) at RT. Then (Boc) ₂ O (16.5 ml; 74.0 mmol) was added dropwise at 0 ° C. The resulting reaction mixture was stirred at RT for 16 h, concentrated under reduced pressure, and water (30 ml) was added to the residue. The crude compound was extracted with ethyl acetate (2x50 ml). The combined organic layer was dried over Na ₂ SO ₄ , filtered and concentrated. The crude material was purified by flash column chromatography (eluting with 2% methanol in DCM) to give an off-white solid (3.1 g; 34%).

M / z 302.36 (M + H) ⁺ .

b) tert-Butyl-5 - [[4- [3,3-bis (tert-butoxycarbonylamino) prop-2 -enoylamino] -3-fluorophenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole- 4-carboxylate.

A solution of tert-butyl 5 - [(4-bromo-3-fluoro-phenyl) sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4 carboxylate (1.1 g; 1.97 mmol) in 1,4- dioxane (15 ml) was purged with argon for 15 min. Then tert-butyl-N- [3-amino-1- (tert-butoxycarbonylamino) -3-oxo-prop-1yl] carbamate (892 mg; 2.95 mmol), K ₃ PO ₄ (837 mg ; 3.94 mmol), Pd ₂ (dba) ₃ (180 mg; 0.19 mmol) and Xantphos (342 mg; 0.59 mmol). The resulting reaction mixture was heated to 65 ° C for 3 h in a sealed tube, cooled to RT, filtered through celite pad and the pad washed with EtOAc (2x10 mL). The filtrate was concentrated under reduced pressure. Received crude so

- 24 038487 the combination was dissolved in ethyl acetate (50 ml), washed with water (30 ml) and brine (30 ml). The organic layer was dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with 55% ethyl acetate in petroleum ether) to give a pale yellow solid (1.3 g; 85%).

M / z 778.52 (M + H) +.

c) 5 - [[4 - [(3-Amino-3-imino-propanoyl) amino] -3-fluoro-phenyl] -sulfonylamino] thiazole-4-carboxylic acid.

TFA (6 ml) was added to tert-butyl-5 - [[4- [3,3-bis (tert-butoxycarbonylamino) prop-2-enoylamino] 3-fluoro-phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate (600 mg; 0.77 mmol) at RT. The resulting mixture was stirred for 3 hours and concentrated under reduced pressure. The resulting crude product was triturated with diethyl ether (2x10 ml) and dried under high vacuum. The crude product was purified by preparative HPLC to give the title product as an off-white solid (47 mg; 15%).

¹ H NMR (400 MHz, DMSO-de) δ 13.42 (1H, brs), 10.34 (1H, brs), 8.99 (2H, brs), 8.62 (2H, brs), 8.14-8.02 (2H, m), 7.58 -7.50 (2H, m), 3.68 (2H, s).

M / z 402.3 (M + H) ⁺ .

LC-MS conditions.

Column: Acquity VEN C18 (50 mm x 2.1 mm; 1.7 μm).

Mobile phase: A: 0.05% formic acid solution in water; B: 0.05% formic acid solution in ACN.

Time (min) /% B: 0/3; 0.4 / 3; 3.2 / 98; 3.8 / 98; 4.2 / 3; 4.5 / 3.

Column temperature: 35 ° C.

Flow rate: 0.6 ml / min.

Preparative HPLC conditions.

Column used: PRONTOSIL (250x19 mm, 5 μm).

Mobile phase: (A): 0.1% formic acid solution, (B): acetonitrile.

Flow rate: 19 ml / min.

Gradient (T /% B): 0/5; 1/5; 7.3 / 59; 7.4 / 99; 11/99; 11.1 / 5; 14/5.

Solubility: ACN + THF + H ₂ O + formic acid.

Examples 5 and 6.

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

Example 6 5 - [[3-carbamoyl-4 - [(2-guanidinoacetyl) amino] phenyl] sulfonylamino] thiazole-4-carboxylic acid.

- 25 038487

a) 4-Bromo-3-cyano-benzenesulfonyl chloride.

Solution A: To a stirred solution of 5-amino-2-bromo-benzonitrile (2 g; 10.1 mmol) in AcOH (25 ml) was added conc. HCl (5 ml) at 0 ° C and stirred for 10 min. Then NaNO2 (770 mg; 11.1 mmol) in H ₂ O (10 ml) was added at the same temperature and stirred for 20 min.

Solution B: Purge AcOH (25 ml) with SO2 gas for 30 min. Then a solution of CuCl ₂ (1.62 g; 12.2 mmol) in H ₂ O (10 ml) was added at 0 ° C and stirred for 20 min. Thereafter, solution B was added dropwise to solution A. The reaction mixture was stirred at RT for 20 min and diluted with water (20 ml). The resulting precipitate was filtered off, washed with n-pentane (2x20 ml) and dried under high vacuum to give a yellow solid (1.7 g; 60%).

b) 5 - [[3-Cyano-4 - [(2-hydroxyacetyl) amino] phenyl] sulfonylmethyl-amino] thiazole-4-carboxylic acid.

This compound was obtained following the procedure outlined in step b) of Example 2.

M / z 658.8 (M + H) +.

c) 5 - [[4 - [(2-Aminoacetyl) amino] -3,5-difluoro-phenyl] sulfonylamino] thiazole-4-carboxylic acid, trifluoroacetate.

This compound was obtained following the procedure outlined in step c of example 2.

M / z 382.4 (M + H) ⁺ .

d) 5 - [[4 - [(2-Aminoacetyl) amino] -3-cyano-phenyl] -sulfonylamino] thiazole-4-carboxylic acid and 5 - [[4 - [(2-aminoacetyl) amino] -3- carbamoyl-phenyl] sulfonylamino] thiazole-4-carboxylic acid.

A mixture of TFA: H2O (9: 1, 5 ml) was added to tert-butyl-5 - [[4 - [[2- (tert-butoxycarbonylamino) acetyl] amino] -3-cyano-phenyl] sulfonyl - [(4 α-methoxyphenyl) methyl] amino] thiazole-4-carboxylate (400 mg; 0.60 mmol) at RT. The reaction mixture was stirred for 6 hours and concentrated under reduced pressure. The resulting material was triturated with diethyl ether (2x10 ml) and dried under high vacuum to give a yellow solid that was used in the next step without further purification (300 mg crude) (72% 5 - [[4 - [(2-aminoacetyl) amino ] -3-cyanophenyl] sulfonylamino] thiazole-4-carboxylic acid and 8% 5 - [[4 - [(2-aminoacetyl) amino] -3-carbamoylphenyl] sulfonylamino] thiazole-4-carboxylic acid).

M / z 382.05 (M + H) + and 400.01 (M + H) +.

f) 5 - [[3-Cyano-4 - [(2-guanidinoacetyl) amino] phenyl] -sulfonylamino] thiazole-4-carboxylic acid and 5 - [[3-carbamoyl-4 - [(2-guanidinoacetyl) amino] phenyl] sulfonylamino] thiazole-4-carboxylic acid.

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Pyrazole-1-carboxamidine (172 mg; 1.18 mmol) and DIPEA (0.3 ml; 1.57 mmol) were added to a stirred solution of 5 - [[4 - [(2-aminoacetyl) amino] -3-cyano- phenyl] sulfonylamino] thiazole-4carboxylic acid and 5 - [[4 - [(2-aminoacetyl) amino] -3-carbamoyl-phenyl] sulfonylamino] thiazole-4carboxylic acid (300 mg; 0.78 mmol) in DMF (5 ml ) at CT. The resulting reaction mixture was stirred at RT for 16 h and concentrated under reduced pressure. Water (5 ml) was added to the residue. The resulting precipitate was filtered off and washed with diethyl ether (2x5 ml). The crude product was purified by preparative HPLC giving the title products:

the compound of example 5 (72 mg; off-white solid)

Ή NMR (400 MHz, DMSO-d ₆ ) δ 13.30 (1H, brs), 10.50 (1H, brs), 8.09 (1H, s), 8.02 (1H, d, J = 2.0 Hz), 7.99 (1H , dd, J = 8.8 Hz, J = 2.0 Hz), 7.82 (1H, d, J = 8.8 Hz), 7.52 (2H, brs), 7.23 (3H, brs), 4.13 (2H, s).

M / z 424.34 (M + H) +;

compound of example 6 (5.2 mg; off-white solid)

Ή NMR (400 MHz, DMSO-d6) δ 13.42 (1H, brs), 12.0 (1H, brs), 8.59 (1H, brs), 8.51 (1H, d), 8.20 (1H, d, J = 2.0 Hz), 8.03 (1H, s), 7.83 (1H, dd, J = 8.8 Hz, J = 2.0 Hz), 7.80 (1H, brs), 7.44 (4H, brs), 4.07 (2H, s ).

M / z 442.34 (M + H) ⁺ .

LC-MS conditions.

Column: Acquity VEN C18 (50 mm x 2.1 mm; 1.7 μm).

Mobile phase: A: 0.05% formic acid solution in water; B: 0.05% formic acid solution in ACN.

Time (min) /% B: 0/3; 0.4 / 3; 3.2 / 98; 3.8 / 98; 4.2 / 3; 4.5 / 3.

Column temperature: 35 ° C.

Flow rate: 0.6 ml / min.

Preparative HPLC conditions.

Column: Symmetry C18 (150x25mm, 10um).

Mobile phase: (A): 0.05% formic acid solution, (B): acetonitrile.

Flow rate: 19 ml / min.

Gradient (T /% B): 0/5; 1/5; 5/20; 10.5 / 24; 10.52 / 99; 12/99; 12.02 / 5; 15/5.

Solubility: ACN + H ₂ O + THF + FA.

Compounds prepared using methods analogous to those described in Examples 2-6 and purified in a similar manner by preparative HPLC are shown in the table below.

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Example Structure Name, NMR data and masses 7 and ^NH / VX ^ N ^ NH, 9 AL about ⁿ O = S ^X ^^ F ε ^ χΝΗ Μγ ° ^Η 0 5 - [[3-Fluoro-4 - [(2guanidinoacetyl) amino] phenyl] sulfonylamino] thiazole-4-carboxylic acid. M / z 415.2 (M-Na) '; ¹ H NMR (d ₆ -DMSO) δ 13.1 (1H, s), 8.08 (1H, s), 7.93 (1H, brs), 7.48 (2H, d, J = 7 Hz), 3.88 (2H, s). eight and ^ΝΗ А Ц1 ο ι S Ν ^Η nA ^ ^oh 0 5 - [[4 - [[2- [Carbamimidoyl (methyl) amino] acetyl] amino] -3-fluorophenyl] sulfonylamino] thiazole-4carboxylic acid. M / z 431.2 (M + H) ⁺ ; ¹ H NMR (d ₆ -DMSO) δ 13.30 (1H, s), 10.14 (1H, brs), 8.44 (1H, s), 8.13 (1H, t), 8.07 (1H, s), 7.57-7.32 (5H , brs), 4.27 (2H, s), 2.95 (3H, s).

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nine ζ ^ ω \ = / o oV ζ-ω ^ΐΟ -Π ^s ' 6 ΙΖ г? 5 - [[2,5-Difluoro-4 - [(2-guanidinoacetyl) amino] phenyl] sulfonylamino] thiazole-4carboxylic acid. M / z 435.3 (M + H) ⁺ ; ¹ H NMR (d ₆ -DMSO) δ 13.3 (1H, s), 10.3 (1H, brs), 8.1 (1H, s), 8.04 (1H, t), 7.57 (2H, t), 7.39-7.09 (4H , brs), 4.10 (2H, s). ten ? L I 0 ^{n n} S ^ / NH Vy ° ^H 0 5 - [[4 - [[2- (4,5-Dihydro-1H-imidazole-2ylamino) acetyl] amino] -2,5-difluorophenyl] sulfonylamino] thiazole-4carboxylic acid. M / z 461.3 (M + H) ⁺ ; ¹ H NMR (d ₆ -DMSO) δ 13.19 (1H, s), 10.1 (1H, brs), 8.39 (2H, brs), 8.10 (1H, s), 8.03 (1H, m), 7.55 (1H, m ), 4.09 (2H, s), 3.60 (4H, s). eleven and 1 ^NH A οΑ VS ⁿ S ^ N N ^^ ^0H O 5 - [[4 - [[(2P) -2-Guanidinopropanoyl] amino] phenyl] sulfonylamino] thiazole-4carboxylic acid. M / z 413.3 (M + H) ⁺ ; ¹ H NMR (d ₆ -DMSO) δ 13.6 (1H, s), 10.3 (1H, s), 8.03 (1H, s), 7.71 (5H, t), 7.256.96 (4H, brs), 4.25 (1H , t, J = 7.2 Hz), 1.40 (ZN, d, J = 6.8 Hz). 12 H ZV ^N W ^NH _n ° iXj> C _c O Ν O = S '^ C ^' F zL ^ XyOI O 5 - [[4 - [[3- (Dimethylamino) -3-iminopropanoyl] amino] -3-fluorophenyl] sulfonylamino] thiazole-4carboxylic acid. M / z 430.3 (M + H) ⁺ ; ¹ H NMR (d ₆ -DMSO) δ 13.5 (1H, brs), 9.51 (1H, brs), 8.45 (1H, t, J = 8 Hz), 8.19 (1 Η, s), 7.91 (2H, m) , 4.22 (2H, s), 3.42 (3H, s), 3.35 (3H, s).

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13 \ = / О О = (ζ-ω- ° “Π ΖΙ J. KZ I 5 - [[4 - [(3-Amino-3-iminopropanoyl) amino] -3,5-difluorophenyl] sulfonylamino] thiazole-4carboxylic acid. Μ / ζ 420.3 (M + H) ⁺ ; ¹ H NMR (d ₆ -DMSO) δ 13.21 (1H, s), 9.01 (2H, brs), 8.58 (2H, brs), 8.12 (1H, s), 7.43 (2H, d, J = 7.2 Hz ), 3.62 (2H, s). fourteen Η ^ΝΗ l «aJ ο ^η 0 = 8 ^ ⁴ ^ S ^ NH νίγθΗ 0 5 - [[6 - [(2-Guanidinoacetyl) amino] pyridazin-3-yl] sulfonylamino] thiazole 4-carboxylic acid. Μ / ζ 401.4 (M + H) ⁺ ; ¹ H NMR (d ₆ -DMSO) δ 13.1 (1H, brs), 11.5 (1H, brs), 8.45 (1H, s), 8.38 (1H, d, J = 9.6 Hz), 8.21 (1H, brs ), 8.10 (1H, s), 8.07 (1H, t), 7.89-7.68 (4H, brs), 4.13 (2H, s). The key intermediate 6-chloro-Zpyridazine sulfonyl chloride was prepared as described in the literature, K. Ashton et al., WO2013 / 123444. 15 ΐ Η ΪΛ Αν “ ^Λ “?. ί I ο ^{Η Η} 0 = 5 S_.NH 0 5 - [[4 - [[2- (4,5-Dihydro-1H-imidazole-2ylamino) acetyl] amino] -3,5-difluorophenyl] sulfonylamino] thiazole-4carboxylic acid. Μ / ζ 461.3 (M + H) ⁺ ; ¹ H NMR (d ₆ -DMSO) δ 13.1 (1H, brs), 10.1 (1H, brs), 8.12 (1H, s), 7.42 (2H, d, J = 9.6 Hz), 4.14 (2H, s ), 3.59 (4H, s).

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16 0 ^{x _{N NH 9. s N H ^ nX}} ^ oh 0 5 - [[4 - [(2-Guanidinoacetyl) amino] -3methoxyphenyl] sulfonylamino] thiazole 4-carboxylic acid. M / z 429.3 (M + H) ⁺ ; ¹ H NMR (d ₆ -DMSO) δ 13.59 (1H, brs), 9.41 (1H, brs), 8.44 (1H, s), 8.10 (1H, d, J = 8.4 Hz), 8.04 (1H, s ), 7.77 (1H, brs), 7.567.32 (4H, brs), 7.30 (2H, t), 4.09 (2H, s), 3.86 (3H, s). The key intermediate 4-acetamido-3-methoxybenzenesulfonyl chloride was prepared following literature procedures described by P. Patel et al., Bioorg. Med. Chern. Lett., 2007, 17, 6610. 17 Cl _u NH 1 NA rAY Υ ^ ν ^λ νη ₂ 0 ^H 0 = 8 ^^ _S NH ^ nX ^ oh 0 5 - [[3-Chloro-4 - [(2-guanidinoacetyl) amino] phenyl] sulfonylamino] thiazole-4carboxylic acid. M / z 433.3 (M + H) ⁺ ; ¹ H NMR (d ₆ -DMSO) δ 13.4 (1H, brs), 9.81 (1H, brs), 8.43 (1H, s), 8.08 (1H, s), 7.95 (1H, d, J = 8.4 Hz ), 7.38 (1H, d, J = 2 Hz), 7.67 (1H, t), 7.51-7.22 (4H, brs), 4.12 (2H, s). The key intermediate 3-chloro-4-nitrobenzene-1-sulfonyl chloride is commercially available.

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eighteen F ^ O _u NH 4 .NH 0 5 - [[4 - [(2-Guanidinoacetyl) amino] -3 (trifluoromethoxy) phenyl] sulfonylamino] thiazole-4-carboxylic acid. M / z 483.0 (M + H) ⁺ ; ¹ H NMR (d ₆ -DMSO) δ 13.38 (1H, s), 10.1 (1H, brs), 8.17 (1H, d, J = 9 Hz), 8.09 (1 Η, s), 7.72 (1H, m) , 7.64 (1H, d, J = 1.5 Hz), 7.54 (1H, brs), 7.39-7.25 (4H, brs), 4.12 (2H, s). The key intermediate 4-bromo-3 (trifluoromethoxy) benzenesulfonyl chloride was prepared following literature procedures, CM. Park et al., J. Med. Chern., 2008, 51, 6902. 19 HH .x, NN ^NH 2 on I Г „iAAr O NH c, NH 0 5 - [[3-Fluoro-4- (3-guanidinopropanoylamino) phenyl] sulfonylamino] thiazole-4-carboxylic acid. M / z 431.2 (M + H) ⁺ ; ¹ H NMR (d ₆ -DMSO) δ 10.19 (1H, s), 8.37 (1H, s), 8.18 (1H, t, J = 8.1 Hz), 7.60 (2H, t), 7.46 (1H, t ), 7.35-6.81 (4H, brs), 3.40 (2H, t), 2.70 (2H, t, J = 6.3 Hz).

Example 20. 5 - [[4 - [(2-Guanidinoacetyl) amino phenyl] sulfonylamino] thiazole-4-carboxylic acid.

a) Ethyl 5 - [[4 - [[2- (tert-butoxycarbonylamino) acetyl] amino] phenyl] sulfonylamino] thiazole-4carboxylate.

DIPEA (0.63 ml; 3.66 mmol) and HATU (696 mg; 1.83 mmol) were added to a stirred solution of 2- (tert-butoxycarbonylamino) acetic acid (321 mg; 1.83 mmol) in DMF (5 ml ). The reaction mixture was stirred at RT for 15 min and then ethyl 5 - [(4aminophenyl) sulfonylamino] thiazole-4-carboxylate (400 mg; 1.22 mmol) was added at the same temperature under N ₂ . The resulting reaction mixture was stirred at RT for 16 h and concentrated under reduced pressure. The resulting crude compound was dissolved in a 10% solution of MeOH in DCM (20 ml), washed with sat. NH ₄ Cl solution (2x10 ml), water (10 ml) and brine (10 ml). The organic layer was dried over Na ₂ SO ₄ , filtered and concentrated in vacuo. The crude material was purified by column chromatography (eluting with 3% MeOH) to give an off-white solid (400 mg; 67%).

M / z 484.8 (M + H) +; 507.06 (M + Na) +.

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b) Ethyl 5 - [[4 - [(2-aminoacetyl) amino] phenyl] sulfonylamino] thiazole-4-carboxylate.

n. a solution of HCl in Et ₂ O (4 ml) was added to ethyl 5 - [[4 - [[2- (tert-butoxycarbonylamino) acetyl] amino] phenyl] sulfonylamino] thiazole-4-carboxylate (400 mg; 0.82 mmol ) in diethyl ether (5 ml) at 0 ° C. The reaction mixture was stirred for 5 h at RT and concentrated under reduced pressure. The crude product was purified by preparative HPLC (HCOOH / CH3CN / H2O) to give an off-white solid (300 mg; 94%).

M / z 385.13 (M + H) +.

c) Ethyl 5 - [[4 - [[2- [[N, N'-bis (tert-butoxycarbonyl) carbamimidoyl] amino] acetyl] amino] phenyl] sulfonylamino] thiazole-4-carboxylate.

DIPEA (0.08 ml; 0.49 mmol) and tert-butyl-N - [(tert-butoxycarbonylamino) -pyrazol-1-ylmethylene] carbamate (87 mg; 0.28 mmol) were added to a stirred solution of ethyl 5- [[4 - [(2aminoacetyl) amino] phenyl] sulfonylamino] thiazole-4-carboxylate (270 mg; 0.70 mmol) in DMF (5 ml) at RT. The resulting reaction mixture was stirred at RT for 16 h and concentrated under reduced pressure. The resulting crude compound was dissolved in a 10% solution of MeOH in DCM (20 ml), washed with water (10 ml) and brine (10 ml). The organic layer was dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The crude material was purified by column chromatography (eluting with 4% MeOH in DCM) to give an off-white solid (250 mg; 56%).

M / z 626.97 (M + H) ⁺ .

d) 5 - [[4 - [[2 - [[(Z) -N, N'-Bis (tert-butoxycarbonyl) carbamimidoyl] amino] acetyl] amino] phenyl] sulfonylamino] thiazole-4-carboxylic acid.

_s NBoc

H y (Af ^N NN HBos ⁿ

0 = 3 ^ I

O

TMSOK (69 mg; 0.54 mmol) was added to a stirred solution of ethyl 5 - [[4 - [[2 - [[N, N'-bis (tert-butoxycarbonyl) carbamimidoyl] amino] acetyl] amino] phenyl] sulfonylamino] thiazole-4-carboxylate (170 mg; 0.27 mmol) in THF (4 ml) at RT under N ₂ . The resulting reaction mixture was stirred at 40 ° C for 5 h and concentrated under reduced pressure. The resulting crude product was triturated with diethyl ether (2x5 ml) and dried under high vacuum. The residue was dissolved in water (5 ml) and acidified with 1N HCl. HCl solution (the pH was adjusted to about 2). The resulting solid was filtered off, washed with n-pentane and dried under high vacuum to give an off-white solid that was used in the next step without further purification (70 mg crude; 43%).

M / z 598.92 (M + H) ⁺ .

f) 5 - [[4 - [(2-Guanidinoacetyl) amino] phenyl] sulfonylamino] thiazole-4-carboxylic acid.

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n. a solution of HCl in ether (1 ml) was added to 5 - [[4 - [[2 - [[(Z) -N, N'-bis (m-tert-butoxycarbonyl) carbamimidoyl] amino] acetyl] amino] phenyl] sulfonylamino] - thiazole-4-carboxylic acid (70 mg; 0.11 mmol) in diethyl ether (2 ml) at 0 ° C. The reaction mixture was stirred at RT for 5 h and concentrated under reduced pressure. The crude product was purified by preparative HPLC to give the title product as an off-white solid (11 mg; 23%).

Ή NMR (500 MHz, DMSO-de) δ 13.59 (1H, s), 10.42 (1H, brs), 8.02 (1H, s), 7.68-7.63 (5H, m), 7.42 (4H, brs), 4.02 ( 2H, s).

M / z 398.78 (M + H) +.

Example 21 5 - [[4 - [[2- (4,5-Dihydro-1H-imidazol-2-yl) acetyl] amino] -3-fluoro-phenyl] sulfonylamino] thiazole-4-carboxylic acid.

a) tert-Butyl 5 - [[4 - [(2-cyanoacetyl) amino] -3-fluoro-phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate.

A solution of 2-cyanoacetic acid (86 mg; 1.01 mmol) and PCl ₅ (210 mg; 1.01 mmol) in DCM (20 ml) was heated to reflux for 30 minutes. The temperature of the reaction mixture was lowered to RT and a solution of tert-butyl 5 - [(4-amino-3-fluoro-phenyl) sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate (500 mg; 1, 01 mmol) in DCM (30 ml) under nitrogen atmosphere. The resulting reaction mixture was heated to reflux for 2.5 h, cooled to RT, diluted with DCM (50 ml) and washed with aqueous NaHCO ₃ (30 ml), water (30 ml) and brine (30 ml). The organic layer was dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with 1-2% MeOH in DCM) to give a pale yellow solid (180 mg; 31%).

M / z 561.43 (M + H) ⁺ .

b) tert-Butyl-5 - [[4 - [[2- (4,5-dihydro-1H-imidazol-2-yl) acetyl] amino] -3-fluoro-phenyl] sulfonyl - [(4methoxyphenyl) methyl] amino] thiazole-4-carboxylate.

RMV OtBu about

HCl gas was passed through a solution of tert-butyl 5 - [[4 - [(2-cyanoacetyl) amino] -3-fluorophenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate (300 mg; 0.53 mmol) in a mixture of ethanol: Et ₂ O (1: 2; 30 ml) at 0 ° С for 2 h.The resulting reaction mixture was kept in a refrigerator for 16 h. Then the volatiles were evaporated under reduced pressure at 40 ° WITH. The residue was dissolved in ethanol (10 ml) and ethylenediamine (48 mg; 0.80 mmol) was added at RT. The reaction mixture was stirred at RT for 16 h and concentrated under reduced pressure. The resulting crude product was triturated with diethyl ether (2x5 ml) and dried under high vacuum to give a pale brown solid which was used in the next step without further purification (330 mg crude).

M / z 548.29 (M-Boc + H) +.

c) 5 - [[4 - [[2- (4,5-Dihydro-1H-imidazol-2-yl) acetyl] amino] -3-fluoro-phenyl] sulfonylamino] thiazole 4-carboxylic acid.

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TFA (3 ml) was added to tert-butyl-5 - [[4 - [[2- (4,5-dihydro-1H-imidazol-2-yl) acetyl] amino] -3-fluorophenyl] sulfonyl - [(4 -methoxyphenyl) methyl] amino] -thiazole-4-carboxylate (300 mg; 0.54 mmol) at RT. The reaction mixture was stirred at RT for 4 h and concentrated under reduced pressure. The resulting crude product was triturated with diethyl ether (2x5 ml) and dried under high vacuum. The crude product was purified by preparative HPLC to give the title product as an off-white solid (26 mg; 11%).

1H NMR (400 MHz, DMSO-d ₆ ) δ 13.50 (1H, brs), 10.30 (1H, brs), 8.29 (2H, brs), 8.08-8.04 (2H, m), 7.547.48 (2H, m) , 3.40 (2H, s), 3.36-3.28 (2H, obs), 2.88-2.85 (2H, m).

M / z 428.37 (M + H) +.

Example 22 5 - [[3 -Fluoro-4 - [[3-imino-3 - (methylamino) propanoyl] amino] phenyl] sulfonylamino] thiazole-4-carboxylic acid.

a) tert-Butyl-5- [[3-fluoro-4- [[3- [hydroxy (methyl) amino] -3-imino-propanoyl] amino] phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate.

MeNHOH-HCl (298 mg; 3.56 mmol) and sodium carbonate (472 mg; 4.45 mmol) were added to a solution of tert-butyl 5 - [[4 - [(2-cyanoacetyl) amino] -3-fluoro- phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate (1 g; 1.78 mmol) in ethanol (15 ml) at RT. The resulting reaction mixture was stirred at 60 ° C for 3 h, cooled to RT, filtered and washed with ethanol (2x10 ml). The combined organic layer was concentrated under reduced pressure. The resulting crude compound was triturated with Et ₂ O (2x10 ml) and dried under high vacuum to give a brown solid which was used in the next step without further purification.

M / z 608.03 (M + H) ⁺ .

b) tert-Butyl-5 - [[3-fluoro-4- [[3-imino-3 - (methylamino) propanoyl] amino] phenyl] sulfonyl- [(4-methoxyphenyl) methyl] amino] thiazole-4- carboxylate.

n $ O HN

Λη ^ ^Ν ~ ΡΜΒ N ^ OiBu

O

Bis (pinacolato) dibor (Adv. Synth. Catal., 2015, 357, 451-462) (357 mg; 1.4 mmol) was added to a solution of tert-butyl-5 - [[3-fluoro-4 - [[3 - [hydroxy (methyl) amino] -3-imino-propanoyl] amino] phenyl] sulfonyl [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate (570 mg; 0.93 mmol) in acetonitrile (10 ml) at CT. The resulting reaction mixture was stirred at RT for 1 h and concentrated under reduced pressure. The crude compound was purified by flash chromatography (eluting with 2% triethylamine in 10% methanol in DCM) to give a pale yellow solid (130 mg; 23%).

M / z 592.05 (M + H) ⁺ .

c) 5 - [[3-Fluoro-4 - [[3-imino-3- (methylamino) propanoyl] amino] phenyl] sulfonylamino] thiazole-4carboxylic acid.

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TFA (3 ml) was added to tert-butyl-5 - [[3-fluoro-4 - [[3-imino-3- (methylamino) propanoyl] amino] phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] -thiazole-4-carboxylate (130 mg; 0.21 mmol) at RT. The reaction mixture was stirred for 2 h at RT and concentrated under reduced pressure. The resulting crude product was triturated with diethyl ether (2x5 ml) and dried under high vacuum. The crude product was purified by preparative HPLC to give the title product as a yellow solid (20 mg; 22%).

¹ H NMR (400 MHz, CF ₃ COOD) δ 9.53 (1H, brs), 8.42 (1H, t, J = 8.0 Hz), 8.20 (1H, s), 7.92 (1H, d, J = 8, 8 Hz), 7.88 (1H, d, J = 9.2 Hz), 4.08 (2H, s), 3.18 (3H, s).

M / z 416.34 (M + H) ⁺ .

LC-MS conditions.

Column: Acquity VEN C18 (50 mm x 2.1 mm; 1.7 μm).

Mobile phase: A: 0.05% formic acid solution in water; B: 0.05% formic acid solution in ACN.

Time (min) /% B: 0/3; 0.4 / 3; 3.2 / 98; 3.8 / 98; 4.2 / 3; 4.5 / 3.

Column temperature: 35 ° C.

Flow rate: 0.6 ml / min.

Preparative HPLC conditions.

Column: Symmetry C18 (300x19mm, 7μm).

Mobile phase: (A): 0.1% formic acid solution, (B): acetonitrile.

Flow rate: 19 ml / min.

Gradient (T /% B): 0/5; 1/5; 8.9 / 40; 8.92 / 99; 12/99; 12.1 / 5; 15/5.

Solubility: ACN + H ₂ O + THF.

Example 23 5 - [[2 - [(2-Guanidinoacetyl) amino] thiazol-5-yl] sulfonylamino] thiazole-4-carboxylic acid.

a) 2-Acetamidothiazole-5-sulfonyl chloride.

N-Thiazol-2-ylacetamide (5 g; 35.2 mmol) was added in portions to a solution of chlorosulfonic acid (11.7 ml; 176 mmol) at 0 ° C. The reaction mixture was stirred at 100 ° C for 4 h, cooled to RT and poured into ice-cold water (100 ml). The resulting precipitate was filtered off and washed with water (20 ml). The precipitate was triturated with n-pentane (2x20 ml) and azeotroped with toluene to give an off-white solid that was used in the next step without further purification (2 g crude; 23%).

M / z 241.23 (M + H) +.

b) Ethyl 5 - [(2-acetamidothiazol-5-yl) sulfonylamino] thiazole-4-carboxylate.

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A solution of ethyl 5-aminothiazole-4-carboxylate (300 mg; 1.74 mmol) in THF (10 ml) was added to a stirred solution of NaH (250 mg; 10.4 mmol) in THF (10 ml) at 0 ° C and stirred for 5 minutes. Then to the reaction mixture was added a solution of 2-acetamidothiazole-5-sulfonyl chloride (502 mg; 2.0 mmol) in THF (10 ml) at 0 ° C. The reaction mixture was stirred at this temperature for 1 h. Ice-cooled water (30 ml) was added to the reaction mixture, which was then washed with EtOAc (2x15 ml). The aqueous layer was acidified to pH 2.0 using 1N hydrochloric acid. HCl solution and extracted with EtOAc (3x15 ml). The organic layer was dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a pale brown solid that was used in the next step without further purification (175 mg crude; 26%).

M / z 377.32 (M + H) +.

c) Ethyl 5 - [(2-aminothiazol-5-yl) sulfonylamino] thiazole-4-carboxylate, hydrochloride.

Concentrated HCl (7 ml) was added to a solution of ethyl 5 - [(2-acetamidothiazole-5yl) sulfonylamino] thiazole-4-carboxylate (700 mg; 1.86 mmol) in ethanol (70 ml) at RT. The reaction mixture was boiled under reflux for 5 h and concentrated under reduced pressure. The resulting crude compound was washed with diethyl ether (20 ml), n-pentane (20 ml) and dried under high vacuum to give a brown solid which was used in the next step without further purification (600 mg crude).

M / z 335.04 (M + H) ⁺ .

d) Ethyl 5 - [[2 - [[2- (tert-butoxycarbonylamino) acetyl] amino] thiazol-5-yl] sulfonylamino] thiazole 4-carboxylate.

_n ^ NHBoc o jHn ° 0 = 5- ^ ⁷

S En ^ X ^ OEt

O

HATU (1.36 g; 3.58 mmol) and DIPEA (2.5 ml; 14.3 mmol) were added to a stirred solution of 2 (tert-butoxycarbonylamino) acetic acid (628 mg; 3.58 mmol) in DMF (6 ml) at CT. The reaction mixture was stirred at RT for 15 min and then ethyl 5 - [(2-aminothiazol-5yl) sulfonylamino] thiazole-4-carboxylate hydrochloride (600 mg; 1.79 mmol) was added at the same temperature under N2. The resulting reaction mixture was stirred at RT for 18 h. Ice-cold water (30 ml) was added and extracted with DCM (3x20 ml). The organic layer was dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The crude compound was purified by flash chromatography (eluting with 60-80% EtOAc in petroleum ether) to give a brown solid (400 mg; 45%).

M / z 492.34 (M + H) ⁺ .

f) 5 - [[2 - [[2- (tert-Butoxycarbonylamino) acetyl] amino] thiazol-5-yl] sulfonylamino] thiazole-4carboxylic acid.

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TMSOK (625 mg; 4.8 mmol) was added to a stirred solution of ethyl 5 - [[2 - [[2- (tert-butoxycarbonylamino) acetyl] amino] thiazol-5-yl] sulfonylamino] thiazole-4-carboxylate (400 mg ; 0.8 mmol) in THF (40 ml) at RT. The reaction mixture was stirred at 40 ° C for 1 h, concentrated under reduced pressure, and water (2 ml) was added to the residue. The reaction mixture was acidified to pH 2 using 1N HCl. HCl solution. The resulting precipitate was filtered off, washed with diethyl ether (2x10 ml), n-pentane (10 ml) and dried under high vacuum to give a pale yellow solid (200 mg; 53%).

M / z 464.30 (M + H) +.

f) 5 - [[2 - [(2-Aminoacetyl) amino] thiazol-5-yl] sulfonylamino] thiazole-4-carboxylic acid, hydrochloride.

Η ^ H ₂

N— _n sA ⁰ o Ϊ N

0 = 5- ^ ⁷ I

S_ / NH ^ nX ^ oh О

A solution of HCl in Et ₂ O (2 M; 10 ml) was added to 5 - [[2 - [[2- (tert-butoxycarbonylamino) acetyl] amino] thiazol-5-yl] sulfonylamino] thiazole-4-carboxylic acid (200 mg; 0.43 mmol) at RT. The reaction mixture was stirred at the same temperature for 3 hours, concentrated under reduced pressure and the resulting residue was washed with diethyl ether (2x5 ml) and n-pentane (5 ml) to give a pale yellow solid which was used in the next step without further purification. (With 150 mg unrefined).

M / z 364.30 (M + H) ⁺ .

g) 5 - [[2 - [(2-Guanidinoacetyl) amino] thiazol-5-yl] sulfonylamino] thiazole-4-carboxylic acid.

DIPEA (0.44 ml; 2.7 mmol) was added to a stirred solution of 5 - [[2 - [(2aminoacetyl) amino] thiazol-5-yl] sulfonylamino] thiazole-4-carboxylic acid hydrochloride (100 mg; 0.27 mmol) and pyrazole-1-carboxamidine hydrochloride (80 mg; 0.55 mmol) in DMF (2 ml) at RT. The reaction mixture was stirred at the same temperature for 6 hours. The DMF was evaporated and then water (3 ml) was added to the resulting crude material with stirring for 5 minutes. The resulting precipitate was filtered off and washed with water (2x2 ml), then dried under high vacuum. The crude compound was purified by preparative HPLC to give an off-white solid (16 mg; 14%).

¹ H NMR (400 MHz, DMSO-d6) δ 13.28 (1H, s), 12.6 (1H, brs), 8.15 (1H, s), 7.71 (1H, s), 7.44 (1H, t, J = 6, 4 Hz), 7.21 (4H, brs), 4.11 (2H, d, J = 6.4 Hz).

M / z 405.9 (M + H) +.

LC-MS conditions.

Column: Acquity VEN C18 (50 mm x 2.1 mm; 1.7 μm).

Mobile phase: A: 0.05% formic acid solution in water; B: 0.05% formic acid solution in ACN.

Gradient: time (min) /% B: 0/3; 0.4 / 3; 3.2 / 98; 3.8 / 98; 4.2 / 3; 4.5 / 3.

Column temperature: 35 ° C.

Flow rate: 0.6 ml / min.

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Preparative HPLC conditions.

Column used: Atlantis T3 (250x19 mm, 5 μm).

Mobile phase: (A): 0.1% formic acid solution, (B): acetonitrile.

Flow rate: 19 ml / min.

Gradient (T /% B): 0/5; 1/5; 8.2 / 55; 8.21 / 99; 10/99; 10.1 / 5; 5/13.

Thinner: ACN + H ₂ O + FA.

Example 24 5 - [[4- [2- (2-Carbamimidoylhydrazino) -2-oxo-ethyl] -3-fluoro-phenyl] sulfonylamino] thiazole-4-carboxylic acid.

a) tert-Butyl 5 - [[4- (2-ethoxy-2-oxo-ethyl) -3-fluoro-phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate.

A mixture of tert-butyl 5 - [(4-bromo-3-fluoro-phenyl) sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4carboxylate (2 g; 3.58 mmol), 3-ethoxy-3- potassium oxo-propanoate (1.2 g; 7.16 mmol) and DMAP (43 mg; 0.35 mmol) in mesitylene (20 ml) were purged with argon gas for 30 min. Then BINAP (2,2'-bis (diphenylphosphino) -1,1'-binaphthyl; 222 mg; 0.35 mmol) and Pd ₂ (dba) ₃ (327 mg; 0.35 mmol) were added at the same temperature. The reaction mixture was stirred at 120 ° C for 18 h, cooled to RT and concentrated under reduced pressure. The crude product was purified by flash chromatography (eluting with 40% EtOAc in petroleum ether) to give a yellow solid which was used in the next step without further purification (0.45 g crude).

M / z 565.43 (M + H) +.

b) tert-Butyl 5 - [[3-fluoro-4- (2-hydrazino-2-oxo-ethyl) phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate.

Hydrazine hydrate (709 mg; 14.1 mmol) was added to a stirred solution of tert-butyl-5 - [[4- (2-ethoxy-2-oxo-ethyl) -3-fluoro-phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate (400 mg; 0.7 mmol) in ethanol (20 ml) at RT. The reaction mixture was refluxed for 5 h and concentrated under reduced pressure to give a brown solid which was used in the next step without further purification (350 mg crude).

M / z 551.42 (M + H) ⁺ .

c) tert-Butyl-5 - [[4- [2- [2-N, N'-bis (tert-butoxycarbonyl) -carbαmimidoyl] hydrazino] -2-oxo-ethyl] 3-fluoro-phenyl] sulfonyl- [ (4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate.

NBoc

NHBoc

O

DIPEA (0.32 ml; 1.89 mmol) was added to a stirred solution of tert-butyl-5 - [[3-fluoro-4- (2-hydrazino-2-oxo-ethyl) phenyl] sulfonyl - [(4-methoxyphenyl) methyl ] amino] thiazole-4-carboxylate (350 mg; 0.63 mmol) and tert-butyl NN - [(tert-butoxycarbonylamino) pyrazol-1-yl-methylene] carbamate (394 mg;

- 39 038487

1.27 mmol) in DMF (5 ml) at RT. The reaction mixture was stirred at the same temperature for 18 hours. Ice-cold water was added to the reaction mixture and stirred for 10 minutes. The resulting precipitate was filtered off, washed with water (2x5 ml) and dried under high vacuum. The crude product was purified by flash chromatography (eluting with 60% EtOAc in petroleum ether) to give a yellow solid (120 mg; 23%).

M / z 793.53 (M + H) +.

d) 5 - [[4- [2- (2-Carbamimidoylhydrazino) -2-oxo-ethyl] -3-fluoro-phenyl] sulfonylamino] thiazole-4carboxylic acid.

TFA (2 ml) was added to tert-butyl-5 - [[4- [2- [2 - [(Z) -N, N'-bis (tert-butoxycarbonyl) carbamimidoyl] hydrazino] -2-oxo-ethyl] -3-fluoro-phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate (120 mg; 0.15 mmol) at RT. The reaction mixture was stirred for 3 hours at the same temperature and concentrated under reduced pressure. The resulting crude material was triturated with diethyl ether (2x5 ml). The crude product was purified by preparative HPLC to give the title product as an off-white solid (23 mg).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 13.40 (1H, brs), 8.06 (1H, s), 7.63 (3H, brs), 7.51-7.39 (3H, m), 3.56 (2H, s).

M / z 417.35 (M + H) ⁺ .

LC-MS conditions.

Column: Acquity VEN C18 (50 mm x 2.1 mm; 1.7 μm).

Mobile phase: A: 0.05% formic acid solution in water; B: 0.05% formic acid solution in ACN.

Time (min) /% B: 0/3; 0.4 / 3; 3.2 / 98; 3.8 / 98; 4.2 / 3; 4.5 / 3.

Column temperature: 35 ° C.

Flow rate: 0.6 ml / min.

Preparative HPLC conditions.

Column used: Symmetry C18 (300x19mm, 7μm).

Mobile phase: (A): 0.05% formic acid solution, (B): acetonitrile.

Flow rate: 19 ml / min.

Gradient (T /% B): 0/2; 1/2; 8/20; 10.5 / 20; 10.51 / 99; 12/99; 12.1 / 2; 15/2.

Solubility: ACN + H ₂ O + THF.

The compound prepared using methods analogous to those described in Examples 20-24 above and purified in a similar manner by preparative HPLC is shown in the table below.

Example Structure Name, NMR data and masses 25 n « ⁿ - ^Н 2 о O = S ^^^ F SA ^H О 5 - [[4- [2 - [(2-Amino-2-imino-ethyl) amino] -2-oxo-ethyl] -3-fluoro-phenyl] sulfonylamino] thiazole-4-carboxylic acid. M / z 416.1 (M + H) ⁺ ; ¹ H NMR (d ₆ -DMSO) δ 13.1 (1H, s), 8.67 (1H, brs), 8.49 (3H, brs), 8.07 (1H, brs), 7.48 (1H, m), 7.43 (2H, m ), 3.89 (2H, s), 3.61 (2H, s), 2.07 (1H, s).

- 40 038487

Example 26 5 - [[3,5-Difluoro-4- (guanidinocarbamoylamino) phenyl] sulfonylamino] thiazole-4carboxylic acid.

a) tert-Butyl 5 - [(4-amino-3,5-difluoro-phenyl) sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4carboxylate.

A saturated solution of NH ₃ in dioxane (120 ml) was added to a mixture of tert-butyl 5 - [(4-bromo-3,5-difluoro-phenyl) sulfonyl - [(4-methoxyphenyl) methyl] amino] -thiazole-4-carboxylate ( 2 g; 3.47 mmol), Xantphos (0.6 g; 1.04 mmol), Pd ₂ (dba) ₃ (0.317 g; 0.34 mmol) and K ₃ PO ₄ (2.2 g; 10, 4 mmol). The resulting mixture was stirred in a sealed tube at 100 ° C for 5 h, filtered through a pad of Celite and the pad was washed with ethyl acetate (2x25 ml). The filtrate was concentrated under reduced pressure. The crude material was purified by flash chromatography (eluted with 50% ethyl acetate in petroleum ether) to give a pale yellow solid (1.25 g; 70%).

M / z 512.4 (M + H) +; 534.56 (M + Na) +.

b) tert-Butyl 5 - [[3,5-difluoro-4 - [(4-nitrophenoxy) carbonylamino] phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate.

(4-Nitrophenyl) carbonochloridate (1.57 g; 7.82 mmol) was added to a stirred solution of tert-butyl 5 - [(4-amino-3,5-difluoro-phenyl) sulfonyl - [(4-methoxyphenyl) methyl] amino ] thiazole-4-carboxylate (2 g; 3.91 mmol) in toluene (120 ml) at room temperature and refluxed for 3 hours. The reaction mixture was concentrated under reduced pressure. The crude product was used in the next step without further purification (3.5 g crude).

c) tert-Butyl 5 - [[4 - [(tert-butoxycarbonylamino) carbamoylamino] -3,5-difluoro-phenyl] sulfonyl [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate.

DIPEA (2.6 ml; 15.5 mmol) was added to the suspension of tert-butyl-5 - [[3,5-difluoro-4 - [(4-nitrophenoxy) carbonylamino] phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino ] thiazole-4-carboxylate (3.5 g; 5.17 mmol) and tert-butyl-N-aminocarbamate (1.36 g; 10.3 mmol) in THF (100 ml) at 0 ° C. The reaction mixture was stirred at RT for 3 h and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with 70% ethyl acetate in petroleum ether) to give a pale yellow solid (1.5 g; 43%).

M / z 670.4 (M + H) +.

d) 5 - [[3,5-Difluoro-4- (hydrazinecarbonylamino) phenyl] sulfonyl- [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylic acid hydrochloride.

- 41 038487

A solution of HCl in Et ₂ O (2 M; 200 ml) was added to tert-butyl-5 - [[4 - [(tert-butoxycarbonylamino) carbamoylamino] -3,5-difluoro-phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate (1.5 g; 2.24 mmol) at RT. The reaction mixture was stirred for 24 h, cooled to 0 ° C over 30 min, and the Et2O was decanted. The crude product was triturated with diethyl ether (2x40 ml) and dried under high vacuum to give an off-white solid (1 g crude).

M / z 514.3 (M + H) ⁺ .

f) 5 - [[4 - [[[N, N'-Bis (tert-butoxycarbonyl) carbamimidoyl] amino] carbamoylamino] -3,5-difluorophenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole -4-carboxylic acid.

NBoc

NHBoc about

DIPEA (3.0 ml; 17.5 mmol) was added to a stirred solution of 5 - [[3,5-difluoro-4 (hydrazinecarbonylamino) phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylic hydrochloride acid (1 g; 1.75 mmol) and tert-butyl- (NZ) -N - [(tert-butoxycarbonylamino) -pyrazol-1-ylmethylene] carbamate (0.54 g; 1.75 mmol) in DMF ( 6 ml) at RT. The reaction mixture was stirred for 5 h and the DMF was removed. Then water was added to the crude product, stirring for 5 minutes. The resulting precipitate was filtered off, washed with water (2x5 ml) and dried under high vacuum to give an off-white solid (1.25 g crude).

M / z 756.1 (M + H) ⁺ .

f) 5 - [[3,5-Difluoro-4- (guanidinocarbamoylamino) phenyl] sulfonylamino] thiazole-4-carboxylic acid.

TFA (13 ml) was added to 5 - [[4 - [[[(Z) -N, N'-bis (tert-butoxycarbonyl) carbamimidoyl] amino] carbamoylamino] -3,5-difluoro-phenyl] sulfonyl - [( 4-methoxyphenyl) methyl] amino] thiazole-4-carboxylic acid (1.25 g; 1.54 mmol) at RT. The reaction mixture was stirred for 3 h at RT and the TFA was evaporated while purging with N2 gas. The resulting crude product was triturated with diethyl ether and purified by preparative HPLC to give the title compound as a white solid (150 mg).

¹ H NMR (300 MHz, DMSO-d ₆ ) δ 13.28 (1H, brs), 8.70 (3H, br s), 8.12 (1H, s), 7.39 (2H, d, J = 6.9 Hz), 7.0 -7.37 (3H, br s).

M / z 436.0 (M + H) +.

LC-MS conditions.

Column: Acquity VEN C18 for ultra high performance liquid chromatography (UPLC) (50 mm x 2.1 mm; 1.7 μm).

Mobile phase: A: 0.1% formic acid solution in water; B: 0.1% formic acid solution in acetonitrile.

Gradient: time (min) /% B: 0/2; 0.2 / 2; 1.5 / 98; 2.6 / 98; 2.61 / 2; 3.2 / 2.

Column temperature: 45 ° C.

Flow rate: 0.8 ml / min.

Preparative HPLC conditions.

Column: X Select C18 (150x30mm, 5um).

Mobile phase: (A): 0.05% formic acid solution in H ₂ O; (B): acetonitrile.

Flow rate: 25 ml / min.

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Gradient (T /% B): 0/50; 8/50; 8/40; 9/40; 9.1 / 98; 11/98; 11.1 / 5; 14/40.

Thinner: ACN + H2O + MeOH + THF.

Example 27 5 - [[3-Fluoro-4- (2-guanidinoethoxycarbonylamino) phenyl] sulfonylamino] thiazole-4carboxylic acid.

a) tert-Butyl 5 - [[3-fluoro-4 - [(4-nitrophenoxy) carbonylamino] phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate.

p-Nitrophenyl chloroformate (1.33 g; 6.0 mmol) was added to a stirred solution of tert-butyl 5 [(4-amino-3-fluoro-phenyl) sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4 β-carboxylate (1 g; 2.02 mmol) in toluene (30 ml) at RT. The resulting reaction mixture was stirred at 120 ° C for 1 hour and concentrated under reduced pressure. The resulting crude product was triturated with n-pentane (2x10 ml) and dried under high vacuum to give an off-white solid that was used in the next step without further purification (1.5 g crude).

M / z 659.43 (M + H) ⁺ .

b) tert-Butyl 5 - [[4- [2- (tert-butoxycarbonylamino) ethoxycarbonylamino] -3-fluoro-phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate.

n Y ^^ NHBoc o / ⁵ η ^ ^Ν ΡΜΒ

O tert-Butyl-N- (2-hydroxyethyl) carbamate (440 mg; 2.73 mmol) and DIPEA (0.97 ml; 5.46 mmol) were added to a stirred solution of tert-butyl-5 - [[3-fluoro -4 - [(4-nitrophenoxy) carbonylamino] phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate (1.2 g; 1.82 mmol) in THF (20 ml) at RT. The resulting reaction mixture was stirred at RT for 2 h. Ice-cold water was added followed by extraction with ethyl acetate (2x50 ml). The combined organic layer was dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluted with 40% ethyl acetate in petroleum ether) to give an off-white solid (500 mg; 40%).

M / z 681.50 (M + H) ⁺ .

c) 5 - [[4- (2-Aminoethoxycarbonylamino) -3-fluoro-phenyl] sulfonylamino] thiazole-4-carboxylic acid, trifluoroacetate.

TFA (5 ml) was added to tert-butyl 5 - [[4- [2- (tert-butoxycarbonylamino) ethoxycarbonylamino] -3fluoro-phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate ( 450 mg; 0.66 mmol) at RT. The reaction mixture was stirred for 24 h at the same temperature and concentrated under reduced pressure. The resulting crude product was triturated with diethyl ether (2x10 ml) and dried under high vacuum to give an off-white solid which was used as follows.

43 038487 step without further purification (350 mg crude).

M / z 405.36 (M + H) ⁺ .

d) 5 - [[3-Fluoro-4- (2-guanidinoethoxycarbonylamino) phenyl] sulfonylamino] thiazole-4-carboxylic acid.

Pyrazole-1-carboxamidine hydrochloride (136 mg; 0.92 mmol) and DIPEA (0.55 ml; 3.0 mmol) were added to a stirred solution of 5 - [[4- (2-aminoethoxycarbonylamino) -3-fluoro-phenyl] sulfonylamino] thiazole-4-carboxylic acid in the form of trifluoroacetate (250 mg; 0.61 mmol) in DMF (6 ml) at RT. The resulting reaction mixture was stirred at RT for 4 h, concentrated under reduced pressure, and water (5 ml) was added to the residue. The resulting precipitate was filtered off, washed with diethyl ether (2x5 ml) and dried under high vacuum. The crude product was purified by preparative HPLC to give the title product as an off-white solid (45 mg; 16%).

Ή NMR (300 MHz, DMSO-d ₆ ) δ 13.42 (1H, brs), 9.60 (1H, s), 8.07 (1H, s), 7.79 (1H, t, J = 8.0 Hz), 7.627.56 (1H, m), 7.51 (1H, d, J = 8.0 Hz, J = 2.0 Hz), 7.46 (1H, dd, J = 10.4 Hz; 2.0 Hz), 7.12 (4H, brs), 4.18 (2H, t, J = 5.2 Hz), 3.48-3.40 (2H, m).

M / z 447.27 (M + H) +.

Compounds prepared using methods analogous to those described in Examples 26 and 27 and purified in a similar manner by preparative HPLC are shown in the table below. ______________________________________________________________________________

Example Structure Name, NMR data and masses 28 About [| ] π p „9. 1 L o NH S ^ NH νίγθΗ O 5 - [[4- (Carbamimidoylcarbamoyl-amino) -3fluoro-phenyl] sulfonylamino] thiazole-4carboxylic acid. M / z 402.9 (M + H) ⁺ ; ¹ H NMR (d ₆ -DMSO) δ 13.5 (1H, brs), 8.18 (1 Η, brs), 8.13 (1H, m), 8.04 (1H, s), 7.72 (1H, brs), 7.40 (2H, m), 6.96-6.61 (4H, m). 29 Η n “ί1ΥΥ ^ΝΗι ς _NH O 5 - [[3-Fluoro-4- (guanidinocarbamoylamino) phenyl] sulfonylamino] thiazole-4carboxylic acid. M / z 416.3 (M + H) ⁺ ; ¹ H NMR (d ₆ -DMSO) δ 8.49 (1H, s), 8.04 (1H, brs), 7.88 (1H, brs), 7.50 (4H, m), 3.94 (2H, brs). thirty Η Η u ^H ο ίΎΥ ^ ^ΝΗ = AL о _S NH 0 5 - [[4 - [(2-Amino-2-iminoethyl) carbamoylamino] -3-fluoro-phenyl] sulfonylamino] thiazole-4-carboxylic acid. M / z 417.3 (M + H) ⁺ ; ¹ H NMR (d ₆ -DMSO) δ 13.5 (1H, brs), 9.01 (1 Η, s), 9.21-8.71 (4H, m), 8.43 (1H, s), 8.19 (1H, t, J = 8 , 4 Hz), 8.06 (1H, s), 7.46 (2H, t, J = 8.8 Hz), 7.29 (1H, m), 4.04 (2H, d, J = 5.2 Hz).

- 44 038487

31 n ^NH _N Ah ₂ О ^Н O = S— sA ^H 0 5 - [[3-Fluoro-4- (2-guanidinoethylsulfanylcarbonylamino) phenyl] sulfonylamino] thiazole-4-carboxylic acid. M / z 463.1 (M + H) ⁺ ; ¹ H NMR (d ₆ -DMSO) δ 13.42 (1H, s), 10.51 (1 Η, s), 8.16 (1H, s), 8.08 (1H, s), 7.79 (1H, t, J = 7.5 Hz), 7.61 (1H, s), 7.51 (2H, m), 7.31-6.79 (4H, m), 3.36 (2H, t, J = 6.5 Hz), 3.36 (2H, t, J = 6, 5 Hz).

Example 32 5 - [[3,5-Difluoro-4- (guanidinocarbamoylamino) phenyl] sulfonylamino] thiazole-4-carboxylic acid.

a) tert-Butyl 5 - [[4 - [(2-chloroacetyl) amino] -3-fluoro-phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate.

Et ₃ N (276 mg; 2.73 mmol) and chloroacetyl chloride (185 mg; 1.64 mmol) were added to a stirred solution of tert-butyl-5 - [(4-amino-3-fluoro-phenyl) sulfonyl - [(4 α-methoxyphenyl) methyl] amino] thiazole-4-carboxylate (450 mg; 0.91 mmol) in DCM (5 ml) at 0 ° C. The resulting reaction mixture was stirred at RT for 2 h, quenched with ice-cold water (5 ml) and extracted with DCM (2x10 ml). The combined organic layer was dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The crude material was purified by trituration with diethyl ether (2x5 ml) to give a green solid which was used in the next step without further purification (400 mg crude).

M / z 570.69 (M + H) +.

b) tert-Butyl 5 - [[4 - [(2-azidoacetyl) amino] -3-fluoro-phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate.

NaN ₃ (92 mg; 1.40 mmol) was added to a stirred solution of tert-butyl-5 - [[4 - [(2chloroacetyl) amino] -3-fluoro-phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino ] amino] thiazole-4-carboxylate (400 mg; 0.70 mmol) in DMF (5 ml) at RT. The resulting reaction mixture was stirred at RT for 16 h, quenched with ice-cold water (10 ml) and extracted with ethyl acetate (2x10 ml). The combined organic extracts were dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The crude material was purified by trituration with diethyl ether (2x5 ml) to give a light brown solid (370 mg; 91%).

M / z 577.23 (M + H) ⁺ .

c) tert-Butyl 5 - [[4 - [(2-aminoacetyl) amino] -3-fluoro-phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate.

- 45 038487

10% Pd / C (300 mg) was added to a solution of tert-butyl-5 - [[4 - [(2-azidoacetyl) amino] -3-fluorophenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] - thiazole-4-carboxylate (370 mg; 0.64 mmol) in EtOAc (10 ml) at RT under nitrogen. The resulting reaction mixture was stirred at RT under a hydrogen atmosphere (balloon pressure) for 16 h, filtered through a Celite pad and washed with EtOAc (20 mL). The filtrate was concentrated under reduced pressure. The crude material was purified by trituration with diethyl ether (2x5 ml) to give a brown solid (340 mg; 96%).

M / z 551.35 (M + H) +.

d) tert-Butyl-5 - [[4 - [[2- (4,5-dihydro-1H-imidazol-2-ylamino) acetyl] amino] -3-fluoro-phenyl] sulfonyl - [(4-methoxyphenyl ) methyl] amino] thiazole-4-carboxylate.

2-Methylsulfanyl-4,5-dihydro-1H-imidazole (124 mg; 0.50 mmol) was added to a stirred solution of tert-butyl-5 - [[4 - [(2-aminoacetyl) amino] -3-fluoro-phenyl ] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate (400 mg; 0.72 mmol) in THF (5 ml) at RT. The resulting reaction mixture was heated to 70 ° C for 48 h in a closed vial and concentrated under reduced pressure to give a yellow solid that was used in the next step without further purification (500 mg crude).

M / z 619.36 (M + H) ⁺ .

f) 5 - [[3,5-Difluoro-4- (guanidinocarbamoylamino) phenyl] sulfonylamino] thiazole-4-carboxylic acid.

TFA (5 ml) was added to tert-butyl-5 - [[4 - [[2- (4,5-dihydro-1H-imidazol-2-ylamino) acetyl] amino] -3fluoro-phenyl] sulfonyl - [(4 α-methoxyphenyl) methyl] amino] thiazole-4-carboxylate (500 mg; 0.50 mmol) at 0 ° C and stirred at RT for 6 h. TFA was evaporated under reduced pressure and the resulting crude product was triturated with diethyl ether (2x5 ml) and dried under vacuum. The crude product was purified by preparative HPLC to give the title product as a white solid (37 mg; 18%).

1H NMR (400 MHz, DMSO-d ₆ ) δ 13.43 (1H, brs), 10.20 (1H, brs), 8.4 (3H, brs), 8.11-8.08 (2H, m), 7.557.48 (2H, m) , 4.08 (2H, s), 3.60 (4H, s).

M / z 443.24 (M + H) +.

LC-MS conditions.

Column: Acquity VEN C18 (50 mm x 2.1 mm; 1.7 μm).

Mobile phase: A: 0.05% formic acid solution in water; B: 0.05% formic acid solution in ACN.

Time (min) /% B: 0/3; 0.4 / 3; 2/98; 3.8 / 98; 4.2 / 3; 4.5 / 3.

Column temperature: 35 ° C.

Flow rate: 0.6 ml / min.

Preparative HPLC conditions.

Column: Atlantis T3 (250x19 mm, 5 μm).

Mobile phase: (A): 0.1% formic acid solution; (B): acetonitrile.

Flow rate: 19 ml / min.

Gradient (T /% B): 0/5; 1/5; 9/30; 10.31 / 99; 12/99; 12.1 / 5; 15/5.

- 46 038487

Solubility: ACN + H2O + THF.

Example 33.

5 - [[3-Fluoro-4 - [[2- (morpholine-4-carboxyimidoylamino) acetyl] amino] phenyl] sulfonylamino] thiazole-4-carboxylic acid.

a) tert-Butyl 5 - [[3-fluoro-4- [[2- (morpholine-4-carbothioylamino) acetyl] amino] phenyl] sulfonyl- [(4methoxyphenyl) methyl] amino] thiazole-4-carboxylate.

A solution of tert-butyl 5 - [[4 - [(2-aminoacetyl) amino] -3-fluoro-phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate (500 mg; 0.9 mmol) and di (imidazol-1-yl) methanethione (242 mg; 1.36 mmol) in CH ₂ Cl ₂ (15 ml) were stirred at RT for 30 min. Then morpholine (118 mg; 1.36 mmol) was added and the resulting reaction mixture was stirred at 40 ° С for 1 h. The reaction mixture was concentrated in vacuo.

The crude compound was purified by flash chromatography eluting with 3% MeOH in CH ₂ Cl ₂ to give a pale pink gummy substance (100 mg; 83%).

M / z 680.42 (M + H) ⁺ .

b) tert-Butyl-5 - [[3-fluoro-4 - [[2- (morpholine-4-carboximidoylamino) acetyl] amino] phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4- carboxylate.

Ag (OTf) (255 mg; 0.99 mmol) was added to a solution of tert-butyl-5 - [[3-fluoro-4 - [[2- (morpholine-4carbothioylamino) acetyl] amino] phenyl] sulfonyl - [(4 -methoxyphenyl) methyl] amino] thiazole-4-carboxylate (450 mg; 0.66 mmol) in a mixture of CH ₂ Cl ₂ : THF (1: 1; 20 ml). The reaction mixture was cooled to -30 ° C and NH ₃ gas was purged for 15 min. The reaction mixture was stirred at RT for 4 h, quenched with MeOH (1 ml) and concentrated under reduced pressure. Added water (25 ml) and was extracted with EtOAc (2x50 ml). The combined organic layer was dried over Na ₂ SO ₄ , filtered and concentrated. The crude material was purified by flash chromatography eluting with 3% MeOH in CH ₂ Cl ₂ to give a black gum (250 mg; 57%).

M / z 663.53 (M + H) ⁺ .

c) 5 - [[3-Fluoro-4 - [[2- (morpholine-4-carboximidoylamino) acetyl] amino] phenyl] sulfonylamino] thiazole-4-carboxylic acid.

A solution of tert-butyl 5 - [[3-fluoro-4 - [[2- (morpholine-4-carboximidoylamino) -acetyl] amino] phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate in a mixture of TFA: H ₂ O (95: 5; 2 ml) was stirred at RT for 3 h. The reaction mixture was concentrated and the crude product was neutralized with methanolic ammonia solution. The reaction mixture was then concentrated under a pony

- 47 038487 female pressure. The crude material was purified by preparative HPLC to give the title product as an off-white solid (12.4 mg; 8%).

1H NMR (400 MHz, DMSO-de) δ 13.32 (1H, brs), 9.67 (1H, s), 8.13 (1H, s), 8.05-7.85 (3H, m), 7.63-7.61 (2H, m), 7.52-7.49 (1H, m), 4.18 (2H, brs), 3.63-3.55 (4H, m), 3.50-3.30 (4H, obs).

M / z 487.34 (M + H) +.

LC-MS conditions.

Column: Acquity VEN C18 (50 mm x 2.1 mm; 1.7 μm).

Mobile phase: A: 0.05% formic acid solution in water; B: 0.05% formic acid solution in ACN.

Time (min) /% B: 0/3; 0.4 / 3; 3.2 / 98; 3.8 / 98; 4.2 / 3; 4.5 / 3.

Column temperature: 35 ° C.

Flow rate: 0.6 ml / min.

Preparative HPLC conditions.

Column used: Symmetry C18 (300x19mm, 7μm).

Mobile phase: (A): 0.1% formic acid solution; (B): acetonitrile.

Flow rate: 19 ml / min.

Gradient (T /% B): 0/5; 1/5; 7.1 / 56; 7.15 / 99; 10/99; 10.1 / 5; 5/13.

Solubility: CH ₃ CN + H ₂ O.

Example 34.

5 - [[4 - [[2 - [(N-Cyanocarbamimidoyl) amino] acetyl] amino] -3-fluoro-phenyl] sulfonylamino] thiazole-4carboxylic acid

DIPEA (0.1 ml; 0.58 mmol) and NaN (CN) 2 (142 mg; 1.6 mmol) were added to a stirred solution of 5 - [[4 - [(2-aminoacetyl) amino] -3-fluoro- phenyl] sulfonylamino] thiazole-4-carboxylic acid (200 mg; 0.53 mmol) in DMF (5 ml) at RT under nitrogen. The reaction mixture was stirred at 50 ° C for 48 h and concentrated under reduced pressure. The crude compound was diluted with water (5 ml) and acidified to pH 2-3 using 1N hydrochloric acid. HCl solution. The resulting precipitate was filtered off and dried under high vacuum.

The crude product was purified by preparative HPLC to give the title product as an off-white solid (20.8 mg; 8%).

Ή NMR (400 MHz, DMSO-d ₆ ) δ 12.8 (1H, brs), 10.0 (1H, s), 8.17 (1H, s), 8.13-8.09 (1H, m), 7.55-7.52 (2H, m) , 6.96 (1H, brs), 6.87 (2H, s), 4.00 (2H, d, J = 6.0 Hz).

M / z 442.18 (M + H) ⁺ .

LC-MS conditions.

Column: Acquity VEN C18 (50 mm x 2.1 mm; 1.7 μm).

Mobile phase: A: 0.05% formic acid solution in water; B: 0.05% formic acid solution in ACN.

Time (min) /% B: 0/3; 0.4 / 3; 2/98; 3.4 / 98; 3.5 / 3; 4/3.

Column temperature: 35 ° C.

Flow rate: 0.6 ml / min.

Preparative HPLC conditions.

Column: XBRIDGE C18 (150x19mm, 5μm).

Mobile phase: (A): 0.1% formic acid solution; (B): acetonitrile.

Flow rate: 19 ml / min.

Gradient (T /% B): 0/0; 2/0; 8/20; 10.9 / 20; 10.95 / 99; 13/99; 13.10 / 0; 16/0.

Solubility: ACN + H ₂ O + THF.

Example 35.

5 - [[4 - [(4-Amino-4-imino-butanoyl) amino] -3-fluoro-phenyl] sulfonylamino] thiazole-4-carboxylic acid.

- 48 038487

a) 5 - [[4- (3-Chloropropanoylamino) -3-fluoro-phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole 4-carboxylic acid.

Acetic anhydride (5 ml) in DCM (5 ml) was added to a stirred solution of 3 chloropropanoyl chloride (1.5 g; 3.04 mmol) in DCM (5 ml) at 0 ° C. After 10 min tert-butyl 5 [(4-amino-3-fluoro-phenyl) sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate (1.5 g) in DCM (10 ml) was added at 0 ° C. The resulting reaction mixture was stirred at RT for 2 h, quenched with ice-cold water (20 ml) and extracted with DCM (2x10 ml). The organic layer was dried over Na ₂ SO ₄ , filtered and concentrated. The crude material was purified by flash chromatography (eluting with 60% ethyl acetate in petroleum ether) to give an off-white solid (600 mg; 33%).

M / z 584.40 (M + H) +.

b) tert-Butyl 5 - [[4- (3-cyanopropanoylamino) -3-fluoro-phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate.

Sodium cyanide (76 mg; 1.54 mmol) was added to a stirred solution of 5 - [[4- (3chloropropanoylamino) -3-fluoro-phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylic acid (600 mg; 1.02 mmol) in DMF (5 ml) at RT. The resulting reaction mixture was stirred at RT for 6 h and quenched with ice-cold water (10 ml). The resulting precipitate was filtered off, washed with Et ₂ O (2x10 ml) and dried under high vacuum to give a brown solid (500 mg; 84%).

M / z 597.24 (M + Na) +.

c) tert-Butyl 5 - [[4 - [(4-amino-4-imino-butanoyl) amino] -3-fluoro-phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate ...

HCl gas was passed through a stirred solution of tert-butyl 5 - [[4- (3cyanopropanoylamino) -3-fluoro-phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate (400 mg; 0, 69 mmol) in a mixture of ethanol: Et ₂ O (1: 4; 10 ml) at 0 ° C for 2 h. The resulting reaction mixture was kept at 4 ° C for 16 h. Then the volatiles were evaporated under reduced pressure. The residue was dissolved in ethanol (5 ml) and NH ₃ gas was bubbled in for 20 min. The volatiles were evaporated under reduced pressure. The resulting crude product was triturated with diethyl ether (2x5 ml) and dried under high vacuum to give a brown solid which was used in the next step without further purification (350 mg crude).

M / z 592.43 (M + H) ⁺ .

d) 5 - [[4 - [(4-Amino-4-imino-butanoyl) amino] -3-fluoro-phenyl] sulfonylamino] thiazole-4-carboxylic acid.

- 49 038487

A mixture of TFA: H2O (95: 5; 3 ml) was added to tert-butyl-5 - [[4 - [(4-amino-4-imino-butanoyl) amino] -3fluoro-phenyl] sulfonyl - [(4-methoxyphenyl ) methyl] amino] thiazole-4-carboxylate (350 mg; 0.592 mmol) at 0 ° C. The resulting reaction mixture was stirred at RT for 6 h and concentrated under reduced pressure. The resulting crude product was triturated with diethyl ether (2x5 ml) and dried under high vacuum. The crude product was purified by preparative HPLC to give the title product as an off-white solid (31 mg; 12%).

1H NMR (400 MHz, DMSO-d ₆ ) δ 13.40 (1H, s), 10.08 (1H, s), 8.99-8.71 (4H, m), 8.07 (1H, s), 8.05-8.01 (1H, m) , 7.54-7.46 (2H, m), 2.85 (2H, t, J = 7.2 Hz), 2.62 (2H, t, J = 7.2 Hz).

M / z 415.93 (M + H) +.

LC-MS conditions.

Column: Acquity VEN C18 (50 mm x 2.1 mm; 1.7 μm).

Mobile phase: A: 0.05% formic acid solution in water; B: 0.05% formic acid solution in ACN.

Time (min) /% B: 0/3; 0.4 / 3; 2/98; 3.4 / 98; 3.5 / 3; 4/3.

Column temperature: 35 ° C.

Flow rate: 0.6 ml / min.

Preparative HPLC conditions.

Column: Symmetry C18 (300x19mm, 7μm).

Mobile phase: (A): 0.1% formic acid solution; (B): acetonitrile.

Flow rate: 19 ml / min.

Gradient (T /% B): 0/5; 1/5; 7/20; 10.1 / 20; 10.1 / 99; 13/99; 13.1 / 5; 16/5.

Solubility: ACN + Н ₂ О + THF + DMSO + conc. FA.

Example 36.

5 - [[4- [3- (4,5-Dihydro-1H-imidazol-2-yl) propanoylamino] -3-fluoro-phenyl] sulfonylamino] thiazole 4-carboxylic acid.

a) 5 - [[4- [3- (4,5-Dihydro-1H-imidazol-2-yl) propanoylamino] -3-fluoro-phenyl] sulfonyl - [(4methoxyphenyl) methyl] amino] thiazole-4-carboxylic acid.

HCl gas was passed through a stirred solution of tert-butyl 5 - [[4- (3cyanopropanoylamino) -3-fluoro-phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate (310 mg; 0, 53 mmol) in a mixture of ethanol: Et ₂ O (1: 4; 15 ml) at 0 ° С for 2 h. The resulting reaction mixture was kept in a refrigerator for 16 h. Then the volatiles were evaporated under reduced pressure. The resulting residue was dissolved in ethanol (5 ml). Then ethylenediamine (32 mg; 0.53 mmol) was added at RT. The resulting reaction mixture was stirred at RT for 8 h and concentrated under reduced pressure. The resulting crude product was triturated with n-pentane (2x5 ml) and dried under high vacuum to give a brown solid which was used in the next step without further purification (400 mg crude).

M / z 618.46 (M + H) ⁺ .

- 50 038487

b) 5 - [[4- [3- (4,5-Dihydro-1H-imidazol-2-yl) propanoylamino] -3-fluoro-phenyl] sulfonylamino] thiazole-4-carboxylic acid.

A mixture of TFA: H ₂ O (95: 5; 3 ml) was added to 5 - [[4- [3- (4,5-dihydro-1H-imidazol-2-yl) propanoylamino] -3fluoro-phenyl] sulfonyl- [ (4-methoxyphenyl) methyl] amino] thiazole-4-carboxylic acid (280 mg; 0.45 mmol) at 0 ° C. The resulting reaction mixture was stirred at RT for 4 h and concentrated under reduced pressure. The resulting crude product was triturated with diethyl ether (2x5 ml) and dried under high vacuum. The crude product was purified by preparative HPLC to give the title product as an off-white solid (21 mg; 10%).

1H NMR (300 MHz, DMSO-d6) δ 13.45 (1H, brs), 9.80 (1H, brs), 8.12-8.04 (2H, m), 7.54-7.44 (2H, m), 3.65 (4H, s), 2.82-2.77 (2H, m), 2.62-2.58 (2H, m).

M / z 441.98 (M + H) +.

LC-MS conditions.

Column: Acquity VEN C18 (50 mm x 2.1 mm; 1.7 μm).

Mobile phase: A: 0.05% formic acid solution in water; B: 0.05% formic acid solution in ACN.

Time (min) /% B: 0/3; 0.4 / 3; 3.2 / 98; 3.8 / 98; 4.2 / 3; 4.5 / 3.

Column temperature: 35 ° C.

Flow rate: 0.6 ml / min.

Preparative HPLC conditions.

Column: Symmetry C18 (300x19mm, 7μm).

Mobile phase: (A): 0.1% formic acid solution; (B): acetonitrile.

Flow rate: 19 ml / min.

Gradient (T /% B): 0/5; 1/5; 7/30; 8.7 / 30; 8.75 / 99; 11/99; 11.1 / 5; 5/13.

Solubility: ACN + H ₂ O + THF.

Example 37.

5 - [[4 - [(3-Amino-3-imino-2-methyl-propanoyl) amino] -3-fluoro-phenyl] sulfonylamino] thiazole-4 carboxylic acid.

a) tert-Butyl 5 - [[4- (2-chloropropanoylamino) -3-fluoro-phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate.

2-Chloropropanoyl chloride (1.48 ml; 15.1 mmol) was added to a stirred solution of tert-butyl-5 [(4-amino-3-fluoro-phenyl) sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4 β-carboxylate (3 g; 6.0 mmol) in DCM (50 ml) at 0 ° C. The resulting reaction mixture was stirred at RT for 2 h and concentrated under reduced pressure. The residue was triturated with diethyl ether (2x50 ml), pentane (2x50 ml) and dried under reduced pressure to give an off-white solid (3.4 g; 95%).

M / z 606.28 (M + Na) +; 582.75 (M-H) ^- .

b) tert-Butyl 5 - [[4- (2-cyanopropanoylamino) -3-fluoro-phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate.

- 51 038487

NaCN (570 mg; 11.6 mmol) was added to a solution of tert-butyl-5 - [[4- (2-chloropropanoylamino) -3-fluorophenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4- carboxylate (3.4 g; 5.8 mmol) in DMF (35 ml) at RT. The resulting reaction mixture was stirred at RT for 16 h and quenched with ice-cold water. The resulting precipitate was filtered off and dried under high vacuum. The crude product was purified by silica gel chromatography (eluting with 40% EtOAc in petroleum ether) to give an off-white solid (1.5 g; 44%).

M / z 597.29 (M + Na) +; 573.62 (M-H) ^- .

c) tert-Butyl-5 - [[3-fluoro-4 - [[3- (hydroxyamino) -3-imino-2-methylpropanoyl] amino] phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate.

NH ₂ OH-HCl (362 mg; 5.22 mmol) and Na ₂ CO ₃ (828 mg; 7.8 mmol) were added to a stirred solution of tert-butyl-5 - [[4- (2-cyanopropanoylamino) -3- fluoro-phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate (1.5 g; 2.61 mmol) in EtOH (30 ml) at RT. The resulting reaction mixture was stirred at 65 ° C for 1 h, cooled to RT, and filtered. The filtrate was concentrated under reduced pressure to give a pale yellow gum which was used in the next step without further purification (1.5 g crude).

M / z 608.48 (M + H) ⁺ .

d) tert-Butyl-5 - [[4 - [(3-amino-3-imino-2-methylpropanoyl) amino] -3-fluoro-phenyl] sulfonyl - [(4methoxyphenyl) methyl] amino] thiazole-4 -carboxylate.

Iron powder (193 mg; 3.4 mmol) was added to tert-butyl-5 - [[3-fluoro-4 - [[3- (hydroxyamino) -3imino-2-methyl-propanoyl] amino] phenyl] sulfonyl- [ (4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate (350 mg; 0.57 mmol) in ethanol: water (1: 1; 3 ml) and heated to reflux for 30 min. Then, 1N hydrochloric acid was added to the reaction mixture. a solution of HCl (0.3 ml) in a mixture of ethanol: water (1: 1; 3 ml) over a period of 30 minutes. The reaction mixture was stirred for another 1 h at 70 ° C, cooled to RT and filtered through celite. The celite pad was washed with ethanol (2x10 ml). The filtrate was concentrated under reduced pressure to give a pale yellow liquid, which was used in the next step without further purification (340 mg crude).

M / z 592.28 (M + H) ⁺ .

f) 5 - [[4 - [(3-Amino-3-imino-2-methyl-propanoyl) amino] -3-fluoro-phenyl] sulfonylamino] thiazole-4carboxylic acid.

Η I oDAAg ^{0 nh} >

VQoh o tert-Butyl-5 - [[4 - [(3-amino-3-imino-2-methyl-propanoyl) amino] -3-fluoro-phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole -4-carboxylate (340 mg; 0.57 mmol) was added to a solution of TFA: H ₂ O (9: 1; 3 ml) at RT. The reaction mixture was stirred at RT for 3 h and concentrated under reduced

- 52 038487 nominal pressure (bath temperature below 30 ° C). The residue was triturated with diethyl ether (2x5 ml) and dried under high vacuum. The crude product was purified by preparative HPLC to give the title product as an off-white solid (48.2 mg).

¹ H NMR (300 MHz, DMSO-d ₆ ) δ 13.40 (1H, brs), 10.25 (1H, s), 8.89 (2H, s), 8.65 (2H, s), 8.11 (1H, s), 8.0 ( 1H, t, J = 8.1 Hz), 7.60-7.50 (2H, m), 3.87 (1H, q, J = 7.2 Hz), 1.49 (3H, d, J = 7.2 Hz).

M / z 416.34 (M + H) +.

LC-MS conditions.

Column: Acquity VEN C18 (50 mm x 2.1 mm; 1.7 μm).

Mobile phase: A: 0.05% formic acid solution in water; B: 0.05% formic acid solution in ACN.

Time (min) /% B: 0/3; 0.4 / 3; 3.2 / 98; 3.8 / 98; 4.2 / 3; 4.5 / 3.

Column temperature: 35 ° C.

Flow rate: 0.6 ml / min.

Preparative HPLC conditions.

Column used: Symmetry C18 (300x19mm, 7μm).

Mobile phase: (A): 0.1% formic acid solution; (B): acetonitrile.

Flow rate: 19 ml / min.

Gradient (T /% B): 0/5; 1/5; 8/50; 8.1 / 99; 11/99; 11.1 / 5; 14/5.

Solubility: ACN + H ₂ O + concentrated FA.

Example 38.

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

a) tert-Butyl-5 - [[3-fluoro-4 - [[2- (2-thioxoimidazolidin-1-yl) acetyl] amino] phenyl] sulfonyl - [(4methoxyphenyl) methyl] amino] thiazole-4-carboxylate ...

A solution of tert-butyl 5 - [[4 - [(2-chloroacetyl) amino] -3-fluoro-phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate (500 mg; 0.87 mmol) in acetonitrile (50 ml) was added to a stirred solution of ethylenediamine (0.29 ml; 4.38 mmol) in acetonitrile (50 ml) over a period of 30 min at 75 ° C. The resulting reaction mixture was stirred at 75 ° C for 2.5 h. Then di (imidazol-1-yl) methanethione (1.56 g; 8.77 mmol) was added at 75 ° C and the reaction mixture was stirred for 1 h at the same temperature, then concentrated under reduced pressure. The resulting crude compound was diluted with EtOAc (50 ml), washed with water (10 ml) and brine (10 ml). The organic layer was dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with 60% EtOAc in petroleum ether) to give a light brown solid (200 mg; 36%).

M / z 636.20 (M + H) ⁺ .

b) tert-Butyl-5 - [[3-fluoro-4 - [[2- (2-iminoimidazolidin-1-yl) acetyl] amino] phenyl] sulfonyl - [(4methoxyphenyl) methyl] amino] thiazole-4-carboxylate ...

Ag (OTf) (121 mg; 0.47 mmol) and saturated NH ₃ in THF (5 ml) were added to a stirred solution of tert-butyl-5 - [[3-fluoro-4 - [[2- (2-thioxoimidazolidine- 1-yl) acetyl] amino] phenyl] sulfonyl - [(4

- 53 038487 methoxyphenyl) methyl] amino] thiazole-4-carboxylate (200 mg; 0.31 mmol) in CH ₂ Cl ₂ (10 ml) at -30 ° C under nitrogen atmosphere. The resulting reaction mixture was stirred at RT for 16 h, quenched with MeOH (2 ml) and stirred at RT for 10 min. The reaction mixture was filtered through a pad of celite and the pad was washed with CH ₂ Cl ₂ (2x10 ml). The filtrate was concentrated under reduced pressure to give a dark brown liquid, which was used in the next step without further purification (300 mg crude).

M / z 619.48 (M + H) ⁺ .

c) 5 - [[3-Fluoro-4 - [[2- (2-iminoimidazolidin-1-yl) acetyl] amino] phenyl] sulfonylamino] thiazole-4carboxylic acid.

A mixture of TFA: H ₂ O (9: 1; 5 ml) was added to tert-butyl-5 - [[3-fluoro-4 - [[2- (2-iminoimidazolidin-1yl) acetyl] amino] phenyl] sulfonyl- [ (4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate (270 mg; 0.43 mmol) at 0 ° C. The resulting reaction mixture was stirred at RT for 3 h and concentrated under reduced pressure. The resulting crude product was triturated with diethyl ether (2x5 ml) and dried under high vacuum. The crude product was purified by preparative HPLC to give the title product as an off-white solid (10.6 mg).

1H NMR (400 MHz, DMSO-d ₆ ) δ 13.32 (1H, brs), 10.24 (1H, s), 8.70 (1H, brs), 8.12 (1H, s), 7.68-7.60 (3H, m), 7.47 (1H, dd, J = 8.0 Hz, J = 7.6 Hz), 7.38 (1H, s), 4.11 (2H, s), 3.74-3.67 (2H, m), 3.61-3.55 (2H, m ).

M / z 443.24 (M + H) +.

LC-MS conditions.

Column: Acquity VEN C18 (50 mm x 2.1 mm; 1.7 μm).

Mobile phase: A: 0.05% formic acid solution in water; B: 0.05% formic acid solution in ACN.

Time (min) /% B: 0/3; 0.4 / 3; 3.2 / 98; 3.8 / 98; 4.2 / 3; 4.5 / 3.

Column temperature: 35 ° C.

Flow rate: 0.6 ml / min.

Preparative HPLC conditions.

Column used: Atlantis T3 (250x19 mm, 5 μm).

Mobile phase: (A): 0.1% formic acid solution; (B): acetonitrile.

Flow rate: 19 ml / min.

Gradient (T /% B): 0/5; 1/5; 7/30, 8.25 / 30; 8.3 / 99; 11/99; 11.1 / 5; 14/5.

Solubility: ACN + H ₂ O + THF.

Example 39.

5 - [[4 - [[2 - [[N- (2-Aminoethyl) carbamimidoyl] amino] acetyl] amino] -3-fluoro-phenyl] sulfonylamino] thiazole-4-carboxylic acid.

a) tert-Butyl-5 - [[4 - [[2- [2- (tert-butoxycarbonylamino) ethylcarbamothioylamino] acetyl] amino] -3fluorophenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate ...

Di (imidazol-1-yl) methanethione (97 mg; 0.54 mmol) was added to a stirred solution of tert

- 54 038487 butyl 5 - [[4 - [(2-aminoacetyl) amino] -3-fluoro-phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4carboxylate (200 mg; 0.36 mmol) in DCM (10 ml) at RT. The reaction mixture was stirred at RT for 4 h and NH ₂ CH ₂ CH ₂ NHBoc (174 mg; 1.08 mmol) was added. The reaction mixture was stirred at 40 ° C for 6 h and concentrated under reduced pressure. The crude product was purified by column chromatography (eluting with 60% EtOAc in petroleum ether) to give a brown solid (80 mg; 29%).

M / z 753.43 (M + H) +.

b) tert-Butyl-5 - [[4 - [[2 - [[N- [2- (tert-butoxycarbonylamino) ethyl] carbamimidoyl] amino] acetyl] amino] -3-fluoro-phenyl] sulfonyl - [( 4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate.

Ag (OTf) (107 mg; 0.47 mmol) was added to a stirred solution of tert-butyl-5 - [[4 - [[2- [2- (tert-butoxycarbonylamino) ethylcarbamothioylamino] acetyl] amino] -3-fluoro-phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate (220 mg; 0.27 mmol) in CH ₂ Cl ₂ (10 ml) at RT. The reaction mixture was stirred at RT for 15 min and saturated NH ₃ in THF (5 ml) was added at -30 ° C under nitrogen atmosphere. The resulting reaction mixture was stirred at RT for 6 h, filtered through a pad of Celite and the pad was washed with CH ₂ Cl ₂ (10 mL). The filtrate was concentrated and the resulting crude material was triturated with n-pentane (10 ml) to give a brown solid which was used in the next step without further purification.

M / z 736.40 (M + H) ⁺ .

c) 5 - [[4 - [[2 - [[N- (2-Aminoethyl) carbamimidoyl] amino] acetyl] amino] -3-fluoro-phenyl] sulfonylamino] thiazole-4-carboxylic acid

A mixture of TFA: H ₂ O (9: 1; 2 ml) was added to tert-butyl-5 - [[4 - [[2 - [[N- [2- (tert-butoxycarbonylamino) ethyl] carbamimidoyl] amino] acetyl]] amino] -3-fluoro-phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate (210 mg; 0.28 mmol) at 0 ° C. The reaction mixture was stirred at RT for 4 h and concentrated under reduced pressure. The resulting crude product was triturated with diethyl ether (2x5 ml) and dried under high vacuum. The crude product was purified by preparative HPLC to give the title product as a brown solid (25 mg).

¹ H NMR (300 MHz, DMSO-d ₆ ) δ 8.13 (1H, brs), 8.09 (1H, s), 7.42-7.30 (2H, m), 6.96 (1H, dd, J = 8.7 Hz, J = 8.4 Hz), 6.20-6.00 (1H, m), 5.70-5.40 (1H, m), 3.82 (2H, s), 3.27 (2H, brs), 2.80-2.75 (2H, m).

M / z 460.30 (M + H) +.

LC-MS conditions.

Column: Acquity VEN C18 (50 mm x 2.1 mm; 1.7 μm).

Mobile phase: A: 0.05% formic acid solution in water; B: 0.05% formic acid solution in ACN.

Time (min) /% B: 0/3; 0.4 / 3; 3.2 / 98; 3.8 / 98; 4.2 / 3; 4.5 / 3.

Column temperature: 35 ° C.

Flow rate: 0.6 ml / min.

Preparative HPLC conditions.

Column: Atlantis T3 (250x19 mm, 5 μm).

Mobile phase: (A): 0.1% formic acid solution; (B): acetonitrile.

Flow rate: 19 ml / min.

Gradient (T /% B): 0/5; 1/5; 7/25; 12/30; 12.1 / 99; 15/99; 15.1 / 5; 18/5.

Solubility: ACN + H ₂ O + THF + FA.

The compound prepared using methods analogous to those described in Example 39 above using methylamine in step a) and purified in a similar manner by preparative HPLC is shown in the table below.

- 55 038487

Example Structure Name, NMR data and masses 40 and ^NH 9 11 o ^{n n} O = S ^^^ F O 5 - [[3-Φτορ-4 - [[2 - [(Ν-methylcarbamimidoyl) amino] acetyl] amino] phenyl] sulfonylamino] thiazole-4carboxylic acid. M / z 431.2 (M + H) ⁺ ; ¹ H NMR (d ₆ -DMSO) δ 13.39 (1H, s), 9.65 (1H, s), 8.11 (1H, m), 8.05 (1H, m), 7.94 (3H, m), 7.61 (2H, m ), 7.49 (1H, t, J = 8.4 Hz), 3.88 (2H, d, J = 5.2 Hz), 2.65 (3H, d, J = 4.4 Hz).

Example 41.

5 - [[4 - [[2- (2-Carbamimidoylhydrazino) acetyl] amino] -3-fluoro-phenyl] sulfonylamino] thiazole-4carboxylic acid.

a) tert-Butyl-5 - [[4 - [[2- (2-tert-butoxycarbonylhydrazino) acetyl] amino] -3-fluoro-phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4 -carboxylate.

KI (1.17 g; 7.01 mmol) was added to a solution of tert-butyl-5 - [[4 - [(2-chloroacetyl) amino] -3-fluorophenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate (2 g; 3.50 mmol) in DMF (20 ml) at RT. After 10 minutes, tert-butyl-Naminocarbamate (695 mg; 5.26 mmol) was added to the reaction mixture at the same temperature. The resulting reaction mixture was stirred at RT for 16 h and concentrated under reduced pressure. Water (25 ml) was added to the crude compound and stirred for 20 minutes. The resulting precipitate was filtered off, washed with diethyl ether and dried under high vacuum to give a pale yellow solid that was used in the next step without further purification (1.2 g; 52%).

M / z 666.48 (M + H) +.

b) 5 - [[3-Fluoro-4 - [(2-hydrazinoacetyl) amino] phenyl] sulfonylamino] thiazole-4-carboxylic acid.

TFA (4 ml) was added to tert-butyl-5 - [[4 - [[2- (2-tert-butoxycarbonylhydrazino) acetyl] amino] -3fluoro-phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole -4-carboxylate (1.2 g; 1.80 mmol) at RT. The reaction mixture was stirred at RT for 16 h and concentrated under reduced pressure. The resulting crude product was triturated with diethyl ether (3x10 ml) to give a pale yellow solid which was used in the next step without further purification (1 g crude).

M / z 390.32 (M + H) ⁺ .

c) 5 - [[4 - [[2- (2-Carbamimidoylhydrazino) acetyl] amino] -3-fluoro-phenyl] sulfonylamino] thiazole-4carboxylic acid.

- 56 038487

DIPEA (1.1 ml; 6.42 mmol) and pyrazole-1-carboxamidine hydrochloride (212 mg; 1.92 mmol) were added to a stirred solution of 5 - [[3-fluoro-4 - [(2-hydrazinoacetyl) amino] phenyl] sulfonylamino] thiazole-4-carboxylic acid (500 mg; 1.28 mmol) in DMF (5 ml) at RT. The resulting reaction mixture was stirred at RT for 16 h, concentrated under reduced pressure, and water (5 ml) was added to the residue. The resulting precipitate was filtered off, washed with Et ₂ O (2x10 ml) and dried under high vacuum. The crude product was purified by preparative HPLC to give the title product as an off-white solid (15 mg).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 13.40 (1H, brs), 10.0 (1H, brs), 9.00 (1H, brs), 8.48 (1H, s), 8.05 (1H, m), 7.55- 7.49 (2H, m), 7.45-7.22 (3H, brs), 5.67 (1H, brs), 3.62 (2H, d, J = 4.4Hz).

M / z 432.37 (M + H) +.

LC-MS conditions.

Column: Acquity VEN C18 (50 mm x 2.1 mm; 1.7 μm).

Mobile phase: A: 0.05% formic acid solution in water; B: 0.05% formic acid solution in ACN.

Time (min) /% B: 0/3; 0.4 / 3; 3.2 / 98; 3.8 / 98; 4.2 / 3; 4.5 / 3.

Column temperature: 35 ° C.

Flow rate: 0.6 ml / min.

Preparative HPLC conditions.

Column: XBRIDGE C18 (150x19mm, 5μm).

Mobile phase: (A): 0.1% formic acid solution; (B): acetonitrile.

Flow rate: 19 ml / min.

Gradient (T /% B): 0/0; 3/0; 8.8 / 33; 9/33; 9.10 / 99; 12/99; 12.10 / 0; 15/0.

Solubility: ACN + H ₂ O + THF + DMSO + F A.

Example 42.

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

a) tert-Butyl 5 - [[4 - [[2- (tert-butoxycarbonylamino) oxyacetyl] amino] -3-fluoro-phenyl] sulfonyl [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate.

A solution of tert-butyl N-hydroxycarbamate (175 mg; 1.31 mmol) in THF (10 ml) was added to a suspension of NaH (195 mg; 4.38 mmol) in THF (10 ml) at 0 ° C under argon and stirred at RT for 30 min. Then, tert-butyl 5 - [[4 - [(2-chloroacetyl) amino] 3-fluoro-phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate (500 mg; 0.87 mmol) in THF (10 ml) at 0 ° C under argon. The resulting reaction mixture was stirred at RT for 1.5 h, quenched with ice-cold water (20 ml) and extracted with ethyl acetate (2x20 ml). The combined organic layer was dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The crude material was purified by column chromatography (eluted with 30% ethyl acetate in petroleum ether) to give a yellow solid (350 mg; 59%).

M / z 667.10 (M + H) ⁺ .

b) 5 - [[4 - [(2-Aminooxyacetyl) amino] -3-fluoro-phenyl] sulfonylamino] thiazole-4-carboxylic acid.

- 57 038487

A mixture of TFA: H2O (9: 1; 3 ml) was added to tert-butyl-5 - [[4 - [[2- (tert-butoxycarbonylamino) oxyacetyl] amino] -3-fluoro-phenyl] sulfonyl - [(4- methoxyphenyl) methyl] amino] thiazole-4-carboxylate (300 mg; 0.44 mmol) at 0 ° C. The reaction mixture was stirred at RT for 4 h and concentrated under reduced pressure. The resulting crude product was triturated with diethyl ether (3x10 ml) to give an off-white solid which was used in the next step without further purification (200 mg crude).

M / z 390.95 (M + H) +.

c) 5 - [[3-Fluoro-4 - [(2-guanidinooxyacetyl) amino] phenyl] sulfonylamino] thiazole-4-carboxylic acid.

DIPEA (0.26 ml; 1.53 mmol) and pyrazole-1-carboxamidine hydrochloride (149 mg; 0.92 mmol) were added to a stirred solution of 5 - [[4 - [(2-aminooxyacetyl) amino] -3-fluoro -phenyl] sulfonylamino] thiazole-4-carboxylic acid (200 mg; 0.51 mmol) in DMF (6 ml) at 0 ° C. The resulting reaction mixture was stirred at RT for 6 h, concentrated under reduced pressure, and water (5 ml) was added to the residue. The resulting precipitate was filtered off, washed with Et ₂ O (2x10 ml) and dried under high vacuum. The crude product was purified by preparative HPLC to give the title product as an off-white solid (70 mg; 31%).

¹ H NMR (300 MHz, DMSO-de) δ 13.50 (1H, brs), 5 9.60 (1H, s), 8.37 (2H, brs), 8.17-8.12 (1H, m), 8.07 (1H, s), 7.58-7.51 (2H, m), 5.45 (2H, brs), 4.62 (2H, brs), 4.22 (2H, s).

M / z 433.30 (M + H) ⁺ .

LC-MS conditions.

Column: Acquity VEN C18 (50 mm x 2.1 mm; 1.7 μm).

Mobile phase: A: 0.05% formic acid solution in water; B: 0.05% formic acid solution in acetonitrile.

Time (min) /% B: 0/3; 0.4 / 3; 3.2 / 98; 3.8 / 98; 4.2 / 3; 4.5 / 3.

Column temperature: 35 ° C.

Flow rate: 0.6 ml / min.

Preparative HPLC conditions.

Column: XBRIDGE C18 (150x19mm, 5μm).

Mobile phase: (A): 0.1% formic acid solution; (B): acetonitrile.

Flow rate: 19 ml / min.

Gradient (T /% B): 0/0; 3/0; 8.8 / 33; 9/33; 9.10 / 99; 12/99; 12.10 / 0; 15/0.

Solubility: ACN + H ₂ O + THF + DMSO + FA.

Example 43 5 - [[4 - [[(2E) -2- (Carbamimidoylhydrazono) acetyl] amino] -3-fluoro-phenyl] sulfonylamino] thiazole-4-carboxylic acid.

a) tert-Butyl-5 - [[4 - [[4- [4 - [(4-tert-butoxycarbonylthiazol-5-yl) - [(4-methoxyphenyl) methyl] sulfamoyl] -2-fluoro-anilino] - 4-oxo-but-2-enoyl] amino] -3-fluoro-phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate.

- 58 038487

But-2-enedioyl dichloride (0.18 g; 1.2 mmol) in DCM (20 ml) was added to a solution of tert-butyl 5 - [(4amino-3-fluoro-phenyl) sulfonyl - [(4-methoxyphenyl ) methyl] amino] thiazole-4-carboxylate (1 g; 2.0 mmol) in DCM (10 ml) at 0 ° C. The resulting reaction mixture was stirred at RT for 6 h, diluted with

DCM and washed with water (2x10 ml) and brine (2x10 ml). The combined organic layer was dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with 50% ethyl acetate in petroleum ether) to give a pale yellow solid (500 mg; 23%).

M / z 1067.05 (M + H) +.

b) tert-Butyl 5 - [[3-fluoro-4- (oxaldehydoylamino) phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate.

A solution of tert-butyl-5 - [[4 - [[4- [4 - [(4-tert-butoxycarbonylthiazol-5-yl) - [(4-methoxyphenyl) methyl] sulfamoyl] -2-fluoro-anilino] -4 -oxo-but-2-enoyl] amino] -3-fluoro-phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate (500 mg; 0.46 mmol) in DCM / MeOH mixture ( 3: 1; 40 ml) was purged with ozone gas for 1 h at -78 ° C. DMS (2 ml) was then added to the reaction mixture at the same temperature and the resulting mixture was stirred at RT for 2 h. The crude reaction mixture was used in the next step without further purification.

c) tert-Butyl 5 - [[4 - [[(2E) -2- (carbamimidoylhydrazono) acetyl] amino] -3-fluoro-phenyl] sulfonyl - [(4 methoxyphenyl) methyl] amino] thiazole-4-carboxylate ...

1-Aminoguanidine hydrochloride (30 mg; 0.27 mmol) was added to a stirred solution of tert-butyl 5 - [[3-fluoro-4- (oxaldehydoylamino) phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4 carboxylate ( 100 mg; 0.18 mmol) in DCM / MeOH (1: 1; 10 ml) at RT. The resulting reaction mixture was stirred at RT for 16 h and concentrated under reduced pressure. The crude product was used in the next step without further purification (100 mg crude).

M / z 522.1 (M + H) ⁺ .

d) 5 - [[4 - [[(2E) -2- (Carbamimidoylhydrazono) acetyl] amino] -3-fluoro-phenyl] sulfonylamino] thiazole 4-carboxylic acid.

A mixture of TFA / H2O (9: 1; 1.0 ml) was added to tert-butyl-5 - [[4 - [[(2E) -2- (carbamimidoylhydrazono) acetyl] amino] -3-fluoro-phenyl] sulfonyl- [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate (80 mg;

- 59 038487

0.13 mmol) at 0 ° C, stirred at RT for 4 h and concentrated under reduced pressure. The resulting crude product was triturated with diethyl ether (2x5 ml) and dried under high vacuum. The crude product was purified by preparative HPLC to give the title product as an off-white solid (11 mg).

1H NMR (400 MHz, DMSO-de) δ 9.68 (1H, brs), 8.05 (1H, s), 7.80 (1H, dd, J = 8.4 Hz, J = 8.0 Hz), 7.537.47 ( 2H, m), 7.19 (1H, s), 6.80-6.00 (4H, brs).

M / z 430.31 (M + H) +.

LC-MS conditions.

Column: Acquity VEN C18 (50 mm x 2.1 mm; 1.7 μm).

Mobile phase: A: 0.05% formic acid solution in water; B: 0.05% formic acid solution in ACN.

Time (min) /% B: 0/3; 0.4 / 3; 3.2 / 98; 3.8 / 98; 4.2 / 3; 4.5 / 3.

Column temperature: 35 ° C.

Flow rate: 0.6 ml / min.

Preparative HPLC conditions.

Column: Atlantis T3 (250x19 mm, 5 μm).

Mobile phase: (A): 0.1% formic acid solution; (B): acetonitrile.

Flow rate: 19 ml / min.

Gradient (T /% B): 0/5; 1/5; 7/25; 12/30; 12.1 / 99; 15/99; 15.1 / 5; 18/5.

Solubility: ACN + H ₂ O + THF + FA.

Example 44 5 - [(4-Guanidinophenyl) sulfonylamino] thiazole-4-carboxylic acid.

n

NH ₂ o || | n „CHA. A NH

0 = 3 ^^^

S SCHN

O

a) tert-Butyl 5 - [[4- [[N, N'-bis (tert-butoxycarbonyl) carbamimidoyl] amino] phenyl] sulfonyl - [(4methoxyphenyl) methyl] amino] thiazole-4-carboxylate.

A solution of tert-butyl 5 - [(4-aminophenyl) sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate (500 mg; 1.05 mmol) in THF (20 ml) was added to a suspension of NaH ( 250 mg; 10.5 mmol) in THF (20 ml) at 0 ° C under argon. After 30 min, a solution of tert-butyl-N - [(tert-butoxycarbonylamino) pyrazol-1-yl-methylene] carbamate (1.0 g; 3.43 mmol) in THF (10 ml) was added at 0 ° C under argon ... The resulting reaction mixture was stirred at RT for 16 h, quenched with ice-cold water (20 ml) and extracted with ethyl acetate (2x20 ml). The combined organic layer was dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure.

The crude material was purified by trituration with diethyl ether (2x5 ml) to give a pale yellow solid that was used in the next step without further purification (150 mg crude).

M / z 662.03 (M + H-Boc) +.

b) 5 - [(4-Guanidinophenyl) sulfonylamino] thiazole-4-carboxylic acid.

n

NH ₂ o ί | I Π ΝΗ

S ^ NH

O

A mixture of TFA: H ₂ O (9: 1; 2 ml) was added to tert-butyl-5 - [[4 - [[N, N'-bis (tert-butoxycarbonyl) carbamimidoyl] amino] phenyl] sulfonyl - [(4 α-methoxyphenyl) methyl] amino] thiazole-4-carboxylate (150 mg; 0.22 mmol) at RT. The resulting mixture was stirred for 4 hours and concentrated under reduced pressure. The resulting crude product was triturated with diethyl ether (2x5 ml) and

- 60 038487 was dried under high vacuum. The crude product was purified by preparative HPLC to give the title product as an off-white solid (22 mg; 28%).

¹ H NMR (400 MHz, DMSO-de) δ 13.60 (1H, brs), 8.05 (1H, s), 7.74 (2H, d, J = 8.8 Hz), 7.47 (3H, brs), 7.25 (2H , d, J = 8.8 Hz).

M / z 342.29 (M + H) +.

LC-MS conditions.

Column: Acquity VEN C18 (50 mm x 2.1 mm; 1.7 μm).

Mobile phase: A: 0.05% formic acid solution in water; B: 0.05% formic acid solution in ACN.

Time (min) /% B: 0/3; 0.4 / 3; 3.2 / 98; 3.8 / 98; 4.2 / 3; 4.5 / 3.

Column temperature: 35 ° C.

Flow rate: 0.6 ml / min.

Preparative HPLC conditions.

Column: Symmetry C18 (300x19mm, 7μm).

Mobile phase: (A): 0.1% formic acid solution; (B): acetonitrile.

Flow rate: 19 ml / min.

Gradient (T /% B): 0/2; 1/2; 8/30; 9.10 / 99; 12/99; 12.10 / 2; 15/2.

Solubility: ACN + H ₂ O + DMSO.

Example 45 5 - [[3-Fluoro-4 - [[(2-guanidinoacetyl) amino] methyl] phenyl] sulfonylamino] thiazole-4-carboxylic acid.

O

a) tert-Butyl 5 - [(3-fluoro-4-vinyl-phenyl) sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate.

about [| o = s ^ I ς / ΊΓ ^pmv . OfRu .OtBu

A solution of tert-butyl 5 - [(4-bromo-3-fluoro-phenyl) sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4 carboxylate (3 g; 5.38 mmol) in 1,4-dioxane ( 40 ml) was purged with argon for 15 min. Then 4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborolane (0.99 g; 6.45 mmol), K ₂ CO ₃ (1.11 g; 8.07 mmol) was added , PdCl ₂ (PPh ₃ ) ₂ (0.37 g; 0.53 mmol) in an argon atmosphere. The resulting reaction mixture was heated to 85 ° C for 24 h in a closed vial. The reaction mixture was allowed to cool to room temperature, filtered through a pad of celite (washed with EtOAc (2x50 ml)). The organic layer was concentrated under reduced pressure. The resulting crude compound was dissolved in ethyl acetate (50 ml), washed with water (50 ml) and brine (50 ml). The organic layer was dried over Na ₂ SO ₄ , filtered and concentrated in vacuo. The crude compound was purified by flash chromatography (eluting with 20% ethyl acetate in petroleum ether) to give an off-white solid (1.5 g; 55%).

M / z 505.1 (M + H) ⁺ .

b) tert-Butyl 5 - [(3-fluoro-4-formyl-phenyl) sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4 carboxylate.

• rmv

OtBu

NaIO ₄ (8.51 g; 39.8 mmol) and OsO ₄ (1.68 g; 6.63 mmol) were added to a solution of tert-butyl-5 - [(3-fluoro4-vinyl-phenyl) sulfonyl - [( 4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate (6.7 g; 13.2 mmol) in acetonutrile: H ₂ O: CCl ₄ (1: 1: 1; 60 ml). The resulting reaction mixture was stirred at RT for 4 h. Water (30 ml) was added and the mixture was extracted with ethyl acetate (2x100 ml). United

- 61 038487 organic extracts were dried over Na ₂ SO ₄ , filtered and concentrated. The crude material was purified by flash chromatography (eluting with 22% ethyl acetate in petroleum ether) to give an off-white solid (4.5 g; 66%).

M / z 507.4 (M + H) +.

c) tert-Butyl 5 - [[3-fluoro-4- [hydroxyiminomethyl] phenyl] sulfonyl- [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate.

о О т O = S '^^' F ς N / ^рмв

O

A solution of tert-butyl 5 - [(3-fluoro-4-formyl-phenyl) sulfonyl - [(4-ethoxyphenyl) methyl] amino] thiazole-4-carboxylate (3.6 g; 7.10 mmol) in EtOH (20 ml ) was added to a stirred solution of hydroxylamine hydrochloride (593.8 mg; 8.52 mmol) and ammonium chloride (454.9 mg; 8.52 mmol) in H ₂ O: EtOH (4: 1; 30 ml). The resulting reaction mixture was stirred at RT for 4 h. Water (20 ml) was added and the mixture was extracted with ethyl acetate (2x50 ml). The combined organic extracts were dried over Na ₂ SO ₄ , filtered and concentrated. The crude material was purified by flash chromatography (eluted with 40% ethyl acetate in petroleum ether) to give an off-white solid (2.8 g; 75%).

M / z 522.1 (M + H) ⁺ .

d) tert-Butyl 5 - [[4- (aminomethyl) -3-fluoro-phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole 4-carboxylate.

Zn (dust; 0.52 g; 8.04 mmol) was added to a stirred solution of tert-butyl-5 - [[3-fluoro-4 [hydroxyiminomethyl] phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole -4-carboxylate (2.8 g; 5.36 mmol) in AcOH (20 ml) at RT. The resulting reaction mixture was stirred at RT for 16 h. Ice-cold water was added and the mixture was extracted with ethyl acetate (2x50 ml). The combined organic extracts were dried over Na2SO4, filtered and concentrated. The crude material was purified by trituration with diethyl ether (2x10 ml) to give a yellow solid (2 g; 73%).

M / z 508.1 (M + H) ⁺ .

f) tert-Butyl-5 - [[4- [[[2- (tert-butoxycarbonylamino) acetyl] amino] methyl] -3-fluoro-phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole- 4-carboxylate.

DIPEA (0.61 ml; 3.54 mmol) and HATU (0.67 g; 1.77 mmol) were added to a solution of tert-butyl-5 - [[4 (aminomethyl) -3-fluoro-phenyl] sulfonyl- [ (4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate (0.6 g; 1.18 mmol) and 2- (tert-butoxycarbonylamino) acetic acid (0.31 g; 1.77 mmol) in DMF (20 ml) in an argon atmosphere. The resulting reaction mixture was stirred at RT for 4 h. Ice-cold water (10 ml) was added and the mixture was extracted with ethyl acetate (2x20 ml). The combined organic extracts were dried over Na ₂ SO ₄ , filtered and concentrated. The crude material was purified by flash chromatography (eluted with 50% ethyl acetate in petroleum ether) to give an off-white solid (500 mg; 63%).

M / z 508.1 (M + H) ⁺ .

f) 5 - [[4 - [[(2-Aminoacetyl) amino] methyl] -3-fluoro-phenyl] sulfonylamino] thiazole-4-carboxylic acid, trifluoroacetate.

- 62 038487

TFA (5 ml) was added to tert-butyl-5 - [[4 - [[[2- (tert-butoxycarbonylamino) acetyl] amino] methyl] -3fluoro-phenyl] sulfonyl - [(4-methoxyphenyl) methyl] amino] thiazole-4-carboxylate (500 mg; 0.75 mmol) at RT and stirred for 4 hours. TFA was evaporated under reduced pressure. The resulting crude product was triturated with diethyl ether (2x10 ml) and dried under high vacuum to give an off-white solid (250 mg; 85%).

M / z 389.1 (M + H) +.

g) 5 - [[3-Fluoro-4 - [[(2-guanidinoacetyl) amino] methyl] phenyl] sulfonylamino] thiazole-4-carboxylic acid.

Pyrazole-1-carboxamidine hydrochloride (141.2 mg; 0.96 mmol) and DIPEA (0.55 ml; 3.21 mmol) were added to a stirred solution of 5 - [[4 - [[(2-aminoacetyl) amino] methyl ] -3-fluorophenyl] sulfonylamino] thiazole-4-carboxylic acid in the form of trifluoroacetate (250 mg; 0.64 mmol) in DMF (6 ml) at RT. The resulting reaction mixture was stirred at RT for 18 h and concentrated under reduced pressure. Water (5 ml) was added to the residue. The resulting precipitate was filtered off and washed with diethyl ether (2x5 ml). The crude product was purified by preparative HPLC to give the title compound as a white solid (70 mg; 25%).

¹ H NMR (500 MHz, DMSO-d6) δ 13.43 (1H, brs), 8.60 (1H, brs), 8.08 (1H, s), 7.51-7.42 (3H, m), 7.507.10 (4H, brs) , 4.33 (2H, s), 3.85 (2H, s).

M / z 431.0 (M + H) ⁺ .

LC-MS conditions.

Column: Acquity UPLC VEN C18 (50 mm x 2.1 mm; 1.7 μm).

Mobile phase: A: 0.1% formic acid solution in water; B: 0.1% formic acid solution in acetonitrile.

Flow rate: 0.8 ml / min.

Time (min) /% B: 0/2; 0.4 / 2; 2.2 / 98; 2.6 / 98; 2.61 / 2; 3.0 / 2.

Column temperature: 60 ° C.

Preparative HPLC conditions.

Column: KROMASIL-C18 (150x25 mm, 10 μm).

Mobile phase: 0.05% formic acid in H ₂ O: acetonitrile.

Flow rate: 25 ml / min.

Gradient (T /% B): 0/5; 1/5; 7/40; 7.1 / 98; 9/98; 9.1 / 5; 11/5.

Solubility: ACN + H ₂ O + THF + DMSO.

The compound prepared using methods analogous to those described in Example 45 above and purified in a similar manner by preparative HPLC is shown in the table below. ______________________________________________________________________________

Example Structure Name, NMR data and masses 46 cm T ZI bA O-W-Z) = o o rh 5 - [[3-Fluoro-4- (guanidinomethyl) phenyl] sulfonylamino] thiazole-4carboxylic acid. M / z 374.0 (M + H) ⁺ ; ¹ H NMR (d ₆ -DMSO) δ 13.40 (1H, s), 8.09 (1H, s), 7.59 (1H, d, J = 7.5 Hz), 7.47 (1H, d, J = 10 Hz), 7.59 (1H, t, J = 7.5 Hz), 7.50-7.25 (4H, brs), 4.41 (2H, s).

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Example 47. Activity of the compounds according to the invention.

Experiments were carried out to determine (1) the inhibitory activity of the compounds of the invention against MBL enzymes;

(2) binding of the compounds of the invention to blood plasma proteins;

(3) the stability of the compounds of the invention in blood plasma.

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

1. Inhibition of enzymes.

In vitro enzyme inhibition assays

Enzyme inhibition assays were performed using solutions of purified MBL enzymes (NDM-1; VIM-1; VIM-2; IMP-1) in 10 mM HEPES (4- (2-hydroxyethyl) -1piperazineethanesulfonic acid) buffer, pH 7, 5 in 96-well microtiter plates. Imipenem (300 μM solution) was used as a substrate and its hydrolysis was monitored at a wavelength of 299 nm in the ultraviolet (UV) range for 10 min every 30 s using a Perkin Elmer Envision UV fluorescence plate reader. The data on the rate of hydrolysis in the presence of a number of inhibitors were analyzed using software from the Dotmatics database and the calculated IC ₅₀ values (concentration causing 50% inhibition) were converted to inhibition constant (Ki) values using the Cheng-Prusoff equation:

where the K _m values for NDM-1, VIM-2 and IMP-1 are 70 μM, 1.5 μM, 9 μM and 25 μM, respectively. Compounds were diluted in DMSO.

Average Ki values obtained in numerous experiments are presented below. Experimental results are shown using the following ranges:

in the case of NDM-1, Ki values less than 0.05 μM are indicated by (A); Ki values in the range of 0.05-0.2 μM are indicated by (B); Ki values greater than 0.2 μM (0.2-2 μM) are indicated by (C);

in the case of VIM-1, Ki values less than 0.2 μM are indicated by (A); Ki values in the 0.2-0.5 μM range are indicated by (B); Ki values greater than 0.5 μM (0.5-1 μM) are indicated by (C);

in the case of VIM-2, Ki values less than 0.02 μM are indicated by (A); Ki values in the range of 0.02-0.05 μM are indicated by (B); Ki values greater than 0.05 μM (0.05-0.15 μM) are indicated by (C);

in the case of IMP-1, Ki values less than 0.5 μM are indicated by (A); Ki values in the range of 0.5-1 μM are indicated by (B); Ki values greater than 1 μM (1-10 μM) are indicated by (C).

Ki values for compounds of the invention

Example Ki (μM) NDM-1 VIM-1 VIM-2 IMP-1 2 (A) (V) (WITH) (WITH) 3 (V) (V) (V) (WITH) 4 (A) (V) (V) (WITH) 5 (V) (A) (V) (A) 6 (WITH) (V) (WITH) 7 (V) (V) (WITH) (WITH) eight (V) (V) (WITH) nine (V) (V) (A) (WITH) ten (A) (A) (A) (V) eleven (WITH) (V) (WITH) 12 (V) (V) (V) 13 (A) (A) (WITH) (V) fourteen (WITH) (V) (WITH) 15 (V) (WITH) (WITH)

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16 (V) (A) (A) (V) 17 (V) (A) (A) (A) eighteen (V) (A) (A) (A) 19 (V) (V) (V) (WITH) twenty (V) (V) 21 (A) (V) (V) 22 (A) (V) (WITH) (WITH) 23 (A) (A) (V) (WITH) 24 (V) (WITH) (WITH) 25 (V) (V) (V) 26 (A) (V) (V) (WITH) 27 (A) (A) (A) (V) 28 (V) (A) (A) (A) 29 (A) (V) (V) (WITH) thirty (A) (V) (V) (WITH) 31 (A) (A) (V) (A) 32 (V) (V) (V) (WITH) 33 (WITH) (V) (WITH) 34 (A) (WITH) (V) (A) 35 (A) (A) (V) (V) 36 (A) (V) (WITH) (WITH) 37 (WITH) (A) (WITH) 38 (WITH) (V) (WITH) 39 (WITH) (A) (V) (WITH) 40 (WITH) (V) (WITH) 41 (A) (V) (V) (V) 42 (WITH) (WITH) (WITH) 43 (V) (V) (A) 44 (WITH) (V) (WITH) (WITH) 45 (WITH) (WITH) (WITH) (WITH) 46 (V) (V) (WITH) (WITH)

2. Antimicrobial susceptibility testing.

Antibiotic activity of β-lactam antibiotics against MBL-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 standardized cationic Mueller-Hinton broth (CAMHB); the concentration range was set from 0.03 to 512 mg / l. Compounds were added at a constant concentration of 8 μg / ml. The turbidity of the bacterial inoculum of each strain (clinical isolates) was adjusted to a 0.5 McFarland turbidity standard in physiological serum (0.9% NaCl), then diluted 1: 100 in CAMHB and added to each well to give a final bacterial cell count of 5x105 CFU / hole. After incubation for 18-20 hours in a heating chamber at 37 ° C., growth inhibition was assessed based on the absence of any bacterial growth.

The minimum inhibitory concentration (MIC) is the lowest antibiotic concentration 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 8 μg / ml. As clinical strains for these drug enhancement experiments, NTBC020 (E. coli strain expressing NDM-1, TEM-1 and CTX-M-15) was used; NTBC035-2 (K. pneumoniae strain expressing NDM-1, CMY-4 and SHV-11); NTBC104-1 (K. pneumoniae strain, express

- 65 038487 NDM-1 and SHV-11); NTBC123 (K. pneumoniae strain expressing NDM-1); NTBC018 (C. freundii strain expressing VIM-2); NTBC024 (K. pneumoniae strain expressing VIM19, TEM-1 and CTX-M-3); NTBC042 (E. coli strain expressing VIM-1, TEM-1, CTX-M-15, SHV-12); NTBC055 (E. coli strain expressing VIM-1); NTBC062 (K. pneumoniae strain expressing IMP-1 and TEM-1) and NTBC039 (K. oxytoca strain expressing IMP-28).

The results are shown below. Data are grouped as follows: MIC values less than 1 μg / ml are indicated by (A); MIC values in the 1-2 μg / ml range are indicated (B); MIC values greater than 2 μg / ml (2-200 μg / ml) are indicated by (C).

Example 48. Comparative study 3. Plasma protein binding.

Brief description of p rotokola Method Rapid equilibrium dialysis Sample Human blood plasma Plasma 100% plasma Test compound concentration 10 μM Buffer Phosphate buffered saline, pH 7.4 Incubation duration 5 h Number of repetitions 2 Quality Control (QC) Connections Warfarin Final concentration of DMSO less than 0.1% Analysis method LC-MS / MS

Analysis method

The test compound was added to blood plasma to give a final concentration of 10 μM. A 300 μL aliquot of plasma was placed in the red chamber of the insert, and 500 μL of PBS (phosphate buffered saline) was placed in the white chamber of the insert. The plate was incubated at

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37 ° C in a thermomixer at 400 rpm for 5 hours. After incubation, the samples were equilibrated in content relative to the content of the opposite sample (10 μl plasma / 100 μl of the buffered sample was compared with 100 μl buffer blank / 10 μl plasma). Samples matched by content were precipitated with 200 μl of acetonitrile containing an internal standard. The samples were vortexed at 1000 rpm for 5 minutes and centrifuged at 4000 rpm for 10 minutes. The supernatant was separated, diluted 2-fold with water and analyzed on an instrument for liquid chromatography combined with tandem mass spectrometry (LC-MS / MS). Blank control samples were processed immediately after the preparation of concentrated working plasma solutions. These samples served as an indicator for calculating the recovery of the test compounds in%.

Counting & Analysis & Data

The percentage of the compound bound to the plasma was calculated in accordance with the following equations:

% unbound compound = 100 * F _c / T _c ,% recovered compound = 100 * (F _c + T _c ) / T ₀ , where

Tc means the total concentration of the compound, determined from the calculated concentration on the plasma side of the membrane;

Fc means the concentration of free compound, determined from the calculated concentration on the membrane from the side of the buffer;

T0 refers to the total compound concentration determined before dialysis.

For each set of duplicates / compound, the percentage of bound compound, the percentage of unbound compound, and the percentage of recovered compound were determined. The results are presented below.

4. Stability of the tested compounds in blood plasma.

Brief description of the protocol

Test compound concentration 1 μM Matrix Human blood plasma Incubation duration 0, 1, 3 and 5 h Number of repetitions 2 for each moment in time Connections for QC Propantheline Analysis method LC-MS / MS End point % Remaining Test Connection

Analysis method

The test compound and the QC compound were incubated at a final concentration of 1 μM in blood plasma at 37 ° C. in a shaking water bath with gentle shaking. At predetermined time points, the reaction was stopped using 200 μl of acetonitrile containing an internal standard, and the reaction mixture was centrifuged at 4000xRCF (relative centrifugal acceleration) and 4 ° C for 20 minutes. The supernatant was separated and analyzed using LC-MS / MS.

Counting & Analysis & Data

The following equation was used to determine the percentage of test compound / compound remaining for QC according to the procedure described above:

I Ratio of peak areas at time (min)% of remaining test. connections —-------------------- h --------------------------- --X 100

Peak area ratio at time 0 min

The results are presented below.

Studies were conducted to compare the compounds of the invention with structurally similar compounds (comp. X). Experiments were carried out as described above. The data is grouped as shown below.

for VIM-1, Ki values less than 0.15 μM are indicated by (++++); Ki values in the range of 0.15-0.3 μM are indicated by (+++); Ki values in the range of 0.3-0.5 μM are indicated by (++); Ki values greater than 0.5 μM (0.5-1 μM) are indicated by (+);

in the case of IMP-1, Ki values less than 0.15 μM are indicated by (++++); Ki values in the range of 0.15-0.6 μM are indicated by (+++); Ki values in the 0.6-5 μM range are indicated by (++); Ki values greater than 5 μM (5-10 μM) are indicated by (+);

in the case of VIM-2, Ki values less than 0.02 μM are indicated by (++++); Ki values in the range of 0.02 - 67 038487

0.05 μM are indicated by (+++); Ki values in the range of 0.05-0.1 μM are indicated by (++); Ki values greater than 0.1 μM (0.1-0.15 μM) are indicated by (+);

in the case of NDM-1, Ki values less than 0.03 μM are indicated by (++++); Ki values in the range of 0.030.1 μM are indicated by (+++); Ki values in the range of 0.1-0.3 μM are indicated by (++); Ki values greater than 0.5 μM (0.3-2 μM) are indicated by (+).

Example Stability in human blood plasma Binding to human blood plasma proteins Conn. A n V ^ NH ₂ F s-Dn O 29% remaining after 2 hours 90.3% Conn. example 7 n ^NH rf ^ Y ^Ν Ν Ν Η ₂ J ^Η O = S SyNH ^ LuON O 100% remaining after 5 hours 60.8% Conn. V ΐ n <Y < ^N > 1 ^ NH2 "HELL S S ^ NH 0 0% remaining after 5 hours No data available Conn. example 2 TH G ii P N NH ₂ ° .1I S ^H O = SF S ^ NH 0 73% remaining after 2 hours 81% Conn. E H s O = S ^^ 0 SyNH ¹ ^ JyOH 0 29% remaining after 2 hours No data available Conn. example 16 H ^NH ^ XA δ ⁿ O = S 0 S ^ NH ¹ ^ Dy ° ⁿ 0 89% remaining after 2 hours No data available

MBL inhibitory effect was observed for the compound of Example 7.

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Example Ki (mkM) VIM-1 IMP-1 VIM-2 NDM-1 Conn. V I n S O = S F S ^ / NH O ++ ++ + ++++ Conn. example 2 F Η ^ΝΗ ι ^^ ΑΚ ^ Ν ^ ΝΙ ^ ^ DL S ⁿ O = SF bDN O ++ ++ ++ ++++ Conn. WITH il'V'lWH; BHP 5 O = S F hs ^ LuON 0 + ++ + ++

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Conn. example 3 u ^NH А if Ύ π ν νη ₂ ? ΑΑ S ⁿ O = SF S ^ / NH ^ ά> ^η 0 ++ ++ +++ ++ Conn. D 0 GU OA ^ CN S ^ / NH 44γ ° ^Η О + +++ +++ +++ Conn. example 5 Η ^ΝΗ [fA ^ ^ JLA S ^н O = S ^{X /} CN S ^ NH ^ Χγ ^0Η 0 +++ +++ +++ +++ Conn. E o ^{gA mng} O = S 0 S ^ / NH ¹ ^ A ^he O ++ ++ ND ++ Conn. example 16 n ^NH rfA ^ ^ AA S ⁿ o = s o S ^ NH ¹ ^ A ^he O ++++ +++ ++++ +++

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Conn. F N S O = S Cl S ^ NH O +++ ND +++ +++ Conn. example 17 Η ^ΝΗ ^ DD S ^h 0 = 5 ^ 01 S ^ NH 0 ++++ ++++ ++++ +++ Conn. G H s 0 = S 0 Ν ^ γ ° Η 0 ++ +++ +++ +++ Conn. example 18 H ^NH ίί ^ γ ^Ν > | ^ Ν ^ ΝΗ ₂ ? -DD S ^H 0-S '0 S _V NH _р ^ _р 0 ++++ ++++ ++++ +++

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Conn. N H <^ < ^N > i ^ NH2 ^ JU s O = S S ^ NH 0 + + +++ ++ Conn. example 20 H ^NH V ^ N ^ NF ^ S ^H o = s S ^ / NH 0 +++ ND ND ++ Conn. 1 ° XT ¹ '' ™ ² “YES o O = SF S ^ NH 0 + ++ ND ++ Conn. example 29 HHW ^H L / U'A and chAD S ⁿ O = SF S ^ / NH 0 +++ ++ +++ ++++

ND: undefined.

It was also found that the compounds of the invention exhibit better enzyme inhibition (lower Ki values) and better enhancement of drug action (lower MIC values) against the aforementioned MBL enzymes (VIM / IMP / NDM) and bacterial strains compared to structurally related analogs lacking the -C (NR) -NR ₂ motif.

As it is easy to see, the compounds according to the invention have improved properties in comparison with structurally analogous compounds. This discovery is surprising, not least because the -C (NR) -NR2 motif common to the compounds of the invention can be subject to rapid hydrolysis, so one would expect these compounds to be unsuitable for applications of the type described in this application. The fact that this motif can be introduced not only without compromising the efficacy of the compounds, but also with increased plasma stability and efficacy, is thus unexpected.

5. Pharmacokinetic (PK) studies.

Compound A and the compound of Example 7 were administered intravenously (iv) at a dose of 1 mg / kg to male Swiss albino mice. The measured PK parameters are shown in the table below.

Compound C _o (n g / ml) AUC (ng h / ml) Cl (ml / min / kg) Compound A 17 nine 819 Compound of Example 7 1814 1036 16

With ₀ means the concentration in blood plasma; AUC means area under the curve; CI stands for Clearance.

It should be noted that in the case of the same dosage

1) when using the compound of example 7, a maximum concentration of more than 100 times higher than that of the compound A is achieved;

2) when using the compound of example 7, an area under the pharmacokinetic curve (AUC, the result of integration of the concentration-time curve) is achieved, more than 100 times higher than that for compound A;

3) the compound of example 7 is cleared from the blood about 50 times slower than compound A.

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These data are consistent with in vitro data obtained for plasma stability, and they confirm that plasma stability is a limiting factor regarding the potential for Compound A to be useful in efficacy studies in animals.

6. In vivo efficacy studies.

Mice were infected by injecting K. pneumoniae NTBC104 into the thigh. The MIC of meropenem for this strain is 64 μg / ml due to the production of NDM-1 by this strain. The MIC of meropenem in the presence of the compound of Example 2 at a concentration of 8 mg / ml is 4 μg / ml.

At the end of the experiment (9 hours after infection), the animals were sacrificed and the number of colony forming units (CFU) was measured to quantify the bacterial load (degree of infection). Meropenem at a dose of 30 mg / kg reduced bacterial load slightly, while the combination of meropenem at a dose of 30 mg / kg with the compound of Example 2 at a dose of 30 mg / kg significantly reduced bacterial load compared to meropenem alone, demonstrating a decrease in CFU to 1.6 log ₁₀ . The results are shown in FIG. 1.

Under the same experimental conditions, the compound of Example 7 caused a decrease in CFU of up to 1.7 log ₁₀ compared to meropenem alone. Compound A was not suitable for efficacy studies because in vitro and PK studies suggested a potential failure when using this compound in efficacy studies, so it would be unethical to conduct such an experiment.

Under the same conditions, the compound of Example 26 caused a decrease in CFU of up to 1.8 log ₁₀ compared to meropenem alone.

7. Expanded MIC Profile Analysis for MBL Expressing Clinical Strains

The susceptibility of about 200 clinical isolates was investigated in order to evaluate the scope and potentiation of the action of meropenem when used in conjunction with the compounds of the invention. The selection criteria for the panel were that the clinical strain should have resistance to carbapenems, but only express NDM variants of the enzyme, and not the serine beta-lactamase enzymes with carbapenemase activity, such as KPC or OXA.

In the case of using the compounds of either example 2 or example 26 at a concentration of 8 μg / ml, the effect of meropenem is enhanced to such an extent that almost 90% of the strains show an MIC for meropenem of 8 μg / ml, while meropenem alone in that the same concentration is able to stop the growth of only less than 1% of the strains, and within the parameters of this experiment, the cessation of growth in 90% of all strains could not be achieved using meropenem alone. The results are shown in FIG. 2.

Example 49.

1. Combination therapy with compounds of the invention together with SBL inhibitors and antibiotics.

As discussed above, bacteria exhibit antibiotic resistance through mechanisms that include both modification of a biological target, resulting in decreased binding affinity for the antibiotic, and the production of enzymes that inactivate the antibacterial drug, such as beta-lactamase enzymes (including serine β-lactamase, SBL, and metal-β-lactamase, MBL). A proposed strategy to address this resistance is to administer a combination therapy containing agents that inhibit antibiotic inactivating enzymes in conjunction with the antibiotic itself. In other words, the antibacterial effect of the drug can be retained by using a dual antibiotic / drug combination approach that inhibits the inactivating enzyme.

Combinations of β-lactamase serine inhibitors with antibiotics are known. For example, a double combination has been developed containing the natural product of streptomycetes clavulanic acid, an inhibitor of serine β-lactamases, together with the β-lactam antibiotic amoxicillin called augmentin. More recently, avibactam, a serine β-lactamase inhibitor with an improved serine β-lactamase inhibition spectrum compared to clavulanic acid in combination with the cephalosporin β-lactam antibiotic ceftazidime (collectively known as avicase), has been introduced to the clinic. However, these combinations are not effective in treating bacterial infections caused by bacteria that express MBL enzymes, since SBL inhibitors are usually inactive against such enzymes.

An additional complication due to the presence of two different categories of β-lactamase enzymes present in the case of bacterial infections arises due to the fact that neither diagnostic tests are currently available in the clinic, allowing very quickly to establish the exact mechanism of β-lactamase resistance, nor possessing double the action of inhibitors of enzymes, both serine and metallo-β-lactamases. Indeed, there is currently no clinically approved metallo-β-lactamase inhibitor that addresses the problem associated with the metallo-β-lactamase enzymes, even if a rapid diagnostic test were available.

- 73 038487 to distinguish between SBL-induced resistance and MBL-induced resistance.

The inventors are now convinced that a pharmaceutical combination of an antibiotic, a serine β-lactamase inhibitor and a metallo-β-lactamase inhibitor (the so-called triple combination) can eliminate the need to identify whether a resistant bacterium causing a particular infection is producing enzyme serine β-lactamase or metallo-β-lactamase (or both, in an increasing number of highly resistant strains). In this regard, there are three possible scenarios for analyzing the β-lactamase profile of carbapenem-resistant enterobacteriaceae (CRE). Microorganisms of group 1 have either exclusively metallo-β-lactamase enzymes or a mixture of metallo-β-lactamase and serine β-lactamase enzymes, but resistance is mainly due to methyl-β-lactamase. Microorganisms of group 2 have only serine β-lactamase enzymes. Group 3 microorganisms have both metallo-β-lactamase and serine β-lactamase enzymes, and both types of enzymes play a significant role in the development of resistance.

This example uses the following abbreviations:

CMY: class C β-lactamase;

TEM: class A β-lactamase;

SHV: class A β-lactamase (with variation for the sulfhydryl group);

CTX-M: class A β-lactamase (CTX for cefotaximase and M for Munich β-lactamase);

OSBL: old spectrum β-lactamases;

OXA: class D β-lactamase (oxacillinase);

ACT-TYPE: class C β-lactamase (AmpC-type beta-lactamase);

KPC: class A β-lactamase (K. pneumoniae carbapenemase);

VIM: Verona integron-encoded metal-β-lactamase;

NDM: New Delhi metallo-β-lactamase;

IMP: metallo-β-lactamase imipenemase.

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 meropenem (mero) were prepared in 96-well plates using standardized cationic Mueller-Hinton broth (CAMHB); the concentration range was set from 0.03 to 512 mg / l. Compounds (the aforementioned compound of Example 2 and / or WCK4234) were added at the concentration indicated in the table below. The turbidity of the bacterial inoculum of each strain (clinical isolates) was corrected to a 0.5 McFarland turbidity standard in physiological serum (0.9% NaCl), then diluted 1: 100 in CAMHB and added to each well, obtaining a final bacterial cell count of 5x10 ⁵ CFU / hole. After incubation for 18-20 hours in a heating chamber at 37 ° C., growth inhibition was assessed based on the absence of any bacterial growth.

The minimum inhibitory concentration (MIC) is the lowest antibiotic concentration 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.

The SBL inhibitor WCK4234 was synthesized according to the procedure described in WO 2015/114595.

WCK4234

M: H (WCK4234) or Na (sodium salt WCK4234).

Briefly, WCK4234 and its sodium salt were synthesized following published procedures (WO 2105114595), the final steps of which are shown below:

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Hydrogenolysis t

Formula (A) sodium salt

NC

Education

OSO ₃ NBu.

Formula (F)

Sulfonation

Formula (E)

The compound of formula (B) was prepared by the synthesis detailed by Ball M. et al. In Organic Process Research and Development (2016), 1799.

The following clinical strains were used in these experiments.

Group 1 (strains, the resistance of which is mainly due to metallo-β-lactamase enzymes):

NTBC020 (E. coli strain expressing NDM-1, TEM-1 and CTX-M-15); NTBC035-2 (K. pneumoniae strain expressing NDM-1, CMY-4 and SHV-11); NTBC104-1 (K. pneumoniae strain expressing NDM-1 and SHV-11); NTBC123 (K. pneumoniae strain expressing NDM-1); NTBC062 (K. pneumoniae strain expressing IMP-1 and TEM-1); NTBC024 (K. pneumoniae strain expressing VIM-19, TEM-1 and CTX-M-3); NTBC042 (E. coli strain expressing VIM-1, TEM-1, CTXM-15, SHV-12); NTBC055 (E. coli strain expressing VIM-1); and NTBC039 (K. oxytoca strain expressing IMP-28).

Group 2 (strains whose resistance is due to serine β-lactamase enzymes):

NTBC091-1 (E. coli strain expressing KPC-2 and TEM-1); NTBC093 (E. cloacae strain expressing KPC-2 and TEM-1); NTBC096-1 (K. pneumoniae strain expressing OXA-181 and SHV11); NTBC099 (K. pneumoniae strain expressing KPC-3, SHV-11 and TEM-1); and NTBC189 (K. pneumoniae strain expressing TEM-OSBL, CTX-M-14 and OXA-48).

Group 3 (strains whose resistance is due to both serine and metallo-β-lactamases):

NTBC019 (K. pneumoniae strain expressing NDM-1, CTX-M-15 and OXA-181); NTBC185 (K. pneumoniae strain expressing SHV-OSBL, TEM-OSBL, NDM-1 and OXA-48); NTBC186 (K. pneumoniae strain expressing ACT-TYPE, VIM-1 and OXA-48); NTBC187 (K. pneumoniae strain expressing SHV-OSBL, NDM-1 and OXA-48); and NTBC188 (K. pneumoniae strain expressing NDM-1 and KPC-2).

The results are presented below. Data are grouped as follows: MIC values less than 1 μg / ml are indicated by (A); MIC values of 1 or 2 μg / ml are indicated (B); MIC values of 4 or 8 μg / ml are indicated (C); and MIC values greater than or equal to 16 μg / ml are indicated by (D).

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As can be seen, for the strains of group 1 and group 2, the use of a double combination of meropenem and the corresponding β-lactamase inhibitor leads to the necessary decrease in MIC.

For microorganisms of group 3, the results indicate that the combination of an MBL inhibitor according to the invention and an SBL inhibitor (WCK4234) together with meropenem is able to reduce the MIC as needed.

1. Discussion.

In the antibacterial field, there is no known example of the use of triple therapy specifically designed to eliminate a bacterial infection. There is one well-known example of the use of triple therapy for the treatment of gastroesophageal reflux disease (GORD), in which H. pylori infection and stomach ulcers are believed to be a component of the disorder, but in this case as directed by the National Institute for Health and Care Excellence ( NICE) recommends treatment with a triple combination of an antiulcer proton pump inhibitor along with two antibiotics (amoxicillin and clarithromycin). Neither in the development nor in the clinic is there any triple combination of antibacterial drugs or antibacterial drugs together with adjuvants such as an inhibitor of β-lactamase enzymes.

One of the important advantages of the proposed triple combination according to the invention is that when faced with the CRE strain and the need to quickly provide treatment, which is essential for the patient's survival, the use of such a triple combination means that, in principle, there is no need to wait for the microbiological and molecular characteristics of the elements resistance before starting treatment. Thus, the triple combination described in this application is useful for preventing or treating any bacterial infection since it eliminates the need for prior identification of the bacterial strain.

2. Other data.

Additional experiments were performed to demonstrate the advantages of the ternary combination of the invention.

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Experiments were carried out as described above. Compounds (compounds of examples 2 and 26) were tested at a concentration of 8 μg / ml. Avibactam and WCK4234 were tested at a concentration of 4 μg / ml. For them, the MIC values were determined.

The tested strains expressed both one carbapenemase from class B (MBL) and one from class A or D (serine beta-lactamase).

NTBC19 is a K. pneumoniae strain expressing NDM-1, CTXM-15 and OXA-181.

NTBC188 is an E. cloacae strain expressing NDM-1 and KPC-2.

Data are grouped as follows: MIC values <0.5 μg / ml are indicated by (A); MIC values in the 1-4 μg / ml range are indicated (B); MIC values> 8 μg / ml (8-512 μg / ml) are indicated by (C). The results are shown in the table below.

NTBC19 NTBC188 Meropenem (WITH) (WITH) Meropenem + avibactam (WITH) (WITH) Meropenem + WCK4234 (WITH) (WITH) Meropenem + compound of example 2 (V) (WITH) Meropenem + compound of example 2 + avibactam (A) (A) Meropenem + compound of example 2 + WCK4234 (A) (A) Meropenem + compound of example 26 (V) (WITH) Meropenem + Example 26 + Avibactam (A) (A) Meropenem + compound of example 26 + WCK4234 (A) (A)

The data clearly show that the use of a triple combination: (1) meropenem; (2) a compound of the invention such as the compound of Example 2 or the compound of Example 26; and (3) an SBL inhibitor such as avibactam or WCK4234 results in a beneficial decrease in MIC values for both strains tested.

Claims (36)

CLAIM 1 (A) is a cyclic group selected from a C ₆ -C ₁₀ aryl group and a 5-10 membered heteroaryl group containing 1, 2 or 3 heteroatoms selected from O, S and N; each R ^{2 is} independently selected from (1) halogen or R ⁸ , where each R ^{8 is} independently selected from CN and —C (O) NR ^f R ^g , where each of R ^f and R ^g is H; 1. A thiazole compound of formula (I) or a pharmaceutically acceptable salt thereof where R ¹ represents H; 2. A compound according to claim 1, wherein ® is a cyclic group selected from a phenyl group and a 5-6 membered heteroaryl group containing 1, 2 or 3 heteroatoms selected from O, S and N. (2) O (C _1-3 alkyl), which may optionally be substituted with 3 halogen substituents; m is 0, 1 or 2; R ³ represents a hydrogen atom; n is 0 or 1; Z is a bond or selected from -NR ¹⁰ C (O) -, -C (O) NR ¹⁰ -, -NR ¹⁰ C (O) NR ¹¹ -, -NR ¹⁰ C (O) O- and -NR ¹⁰ C (O) S-; L is a bond or is C _1-3 alkylene; or L is -C (R ¹⁰ ) = N-; - 77 038487 X is a bond or, if L is different from a bond or —C (R ¹⁰ ) = N—, then X is a bond or is selected from —NR ¹⁰ -, —O— and —C (NR ¹⁰ ) -; p is 0 or 1; A i) R ⁴ is H and R ^{5 is} selected from H, —CN, and C 1 -C ₃ alkyl, which is unsubstituted or substituted with —NR ¹⁰ R ¹¹ ; or R ⁴ together with R ⁵ forms, together with the atoms to which they are attached, a 5-6 membered heterocyclic group containing at least one saturated ring carbon atom and containing 1 or 2 heteroatoms selected from O, S and N ; and ii) R ^{6 is} selected from H, —CN, and C1-C3 alkyl, which is unsubstituted or substituted with —NR ¹⁰ R ¹¹ ; and R ⁷ , if present, is selected from H, —CN, and C1-C3 alkyl; or R ⁶ together with R ⁷ , if present, forms, together with the atoms to which they are attached, a 5-6 membered heterocyclic group containing at least one saturated ring carbon atom and containing 1 or 2 heteroatoms selected from O, S and N; or V R ⁴ represents H; R ⁵ together with R ⁶ forms, together with the atoms to which they are attached, a 5-6 membered heterocyclic group containing at least one saturated ring carbon atom and containing 1 or 2 heteroatoms selected from O, S and N; and R ^7, if present, is selected from H, -CN and _C1-C3 alkyl; R ¹⁰ and R ¹¹ are each H. 3. A compound according to any one of claims 1, 2, wherein ® is selected from phenyl, pyridazine, pyridine and thiazole. 4. A compound according to any one of claims 1 to 3, wherein (R) is phenyl. 5 - [[4 - [(2-guanidinoacetyl) amino] phenyl] sulfonylamino] thiazole-4-carboxylic acid; and its pharmaceutically acceptable salts. 5 - [[3-fluoro-4 - [(2-guanidinoacetyl) amino] phenyl] 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-fluorophenyl] sulfonylamino] thiazole-4-carboxylic acid; 5 - [[3-fluoro-4- (3-guanidinopropanoylamino) phenyl] sulfonylamino] thiazole-4-carboxylic acid; 5 - [[5-fluoro-6 - [(2-guanidinoacetyl) amino] -3-pyridyl] sulfonylamino] thiazole-4-carboxylic acid; 5 - [[4 - [[2 - [[N- (2-aminoethyl) carbamimidoyl] amino] acetyl] amino] -3-fluorophenyl] sulfonylamino] thiazole-4-carboxylic acid; 5 - [[4 - [[2- [carbamimidoyl (methyl) amino] acetyl] amino] -3-fluorophenyl] sulfonylamino] thiazole-4carboxylic acid; 5 - [[3-fluoro-4 - [[2- (2-iminoimidazolidin-1-yl) acetyl] amino] phenyl] sulfonylamino] thiazole-4 carboxylic acid; 5 - [[3-fluoro-4 - [[2 - [(N-methylcarbamimidoyl) amino] acetyl] amino] phenyl] sulfonylamino] thiazole-4carboxylic acid; 5 - [[2,5-difluoro-4 - [(2-guanidinoacetyl) amino] phenyl] 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; - 79 038487 5 - [[4 - [(4-amino-4-imino-butanoyl) amino] -3-fluorophenyl] sulfonylamino] thiazole-4-carboxylic acid; 5 - [[3,5-difluoro-4 - [(2-guanidinoacetyl) amino] phenyl] sulfonylamino] thiazole-4-carboxylic acid; 5 - [[4 - [[(2R) -2-guanidinopropanoyl] amino] phenyl] sulfonylamino] thiazole-4-carboxylic acid; 5 - [[4- (carbamimidoylcarbamoylamino) -3-fluorophenyl] sulfonylamino] thiazole-4-carboxylic acid; 5 - [[4 - [[2- (4,5-Dihydro-1H-imidazol-2-yl) acetyl] amino] -3-fluorophenyl] sulfonylamino] thiazole-4carboxylic acid; 5 - [[4 - [(3-amino-3-imino-2-methyl-propanoyl) amino] -3-fluorophenyl] sulfonylamino] thiazole-4 carboxylic acid; 5 - [[3-fluoro-4 - [[2- (morpholine-4-carboximidoylamino) acetyl] amino] phenyl] sulfonylamino] thiazole-4-carboxylic acid; 5 - [[3-fluoro-4- (guanidinocarbamoylamino) phenyl] sulfonylamino] thiazole-4-carboxylic acid; 5 - [[4 - [[2 - [(N-cyanocarbamimidoyl) amino] acetyl] amino] -3-fluorophenyl] sulfonylamino] thiazole-4carboxylic acid; 5 - [[2 - [(2-guanidinoacetyl) amino] thiazol-5-yl] sulfonylamino] thiazole-4-carboxylic acid; 5 - [[4- [3- (4,5-dihydro-1H-imidazol-2-yl) propanoylamino] -3-fluorophenyl] sulfonylamino] thiazole-4carboxylic acid; 5 - [[3-fluoro-4 - [[3-imino-3 - (methylamino) propanoyl] amino] phenyl] sulfonylamino] thiazole-4 carboxylic acid; 5 - [[3-fluoro-4 - [(2-guanidinooxyacetyl) amino] phenyl] sulfonylamino] thiazole-4-carboxylic acid; 5 - [[4 - [[3- (dimethylamino) -3-iminopropanoyl] amino] -3-fluorophenyl] sulfonylamino] thiazole-4 carboxylic acid; 5 - [[4 - [(3-amino-3-iminopropanoyl) amino] -3,5-difluorophenyl] sulfonylamino] thiazole-4-carboxylic acid; 5 - [[3,5-difluoro-4- (guanidinocarbamoylamino) phenyl] sulfonylamino] thiazole-4-carboxylic acid; 5 - [[4 - [(2-amino-2-iminoethyl) carbamoylamino] -3-fluorophenyl] sulfonylamino] thiazole-4-carboxylic acid; 5 - [[6 - [(2-guanidinoacetyl) amino] pyridazin-3-yl] 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 - [[4 - [[(2E) -2- (carbamimidoylhydrazono) acetyl] amino] -3-fluorophenyl] sulfonylamino] thiazole-4carboxylic acid; 5 - [[4 - [[2- (2-carbamimidoylhydrazino) acetyl] amino] -3-fluorophenyl] sulfonylamino] thiazole-4carboxylic acid; 5 - [[4 - [(2-guanidinoacetyl) amino] -3-methoxyphenyl] sulfonylamino] thiazole-4-carboxylic acid; 5 - [[3-chloro-4 - [(2-guanidinoacetyl) amino] phenyl] sulfonylamino] thiazole-4-carboxylic acid; 5 - [[4- [2- (2-carbamimidoylhydrazino) -2-oxoethyl] -3-fluorophenyl] sulfonylamino] thiazole-4carboxylic acid; 5 - [(4-guanidinophenyl) sulfonylamino] thiazole-4-carboxylic acid; 5 - [[3-fluoro-4- (2-guanidinoethoxycarbonylamino) phenyl] sulfonylamino] thiazole-4-carboxylic acid; 5 - [[3-cyano-4 - [(2-guanidinoacetyl) amino] phenyl] sulfonylamino] thiazole-4-carboxylic acid; 5 - [[3-carbamoyl-4 - [(2-guanidinoacetyl) amino] phenyl] sulfonylamino] thiazole-4-carboxylic acid; 5 - [[4- [2 - [(2-amino-2-iminoethyl) amino] -2-oxoethyl] -3-fluorophenyl] sulfonylamino] thiazole-4carboxylic acid; 5 - [[3-fluoro-4- (2-guanidinoethylsulfanylcarbonylamino) phenyl] sulfonylamino] thiazole-4carboxylic acid; 5 - [[3-fluoro-4- (guanidinomethyl) phenyl] sulfonylamino] thiazole-4-carboxylic acid; 5 - [[3-fluoro-4 - [[(2-guanidinoacetyl) amino] methyl] phenyl] sulfonylamino] thiazole-4-carboxylic acid; 5 - [[4 - [(2-guanidinoacetyl) amino] -3 - (trifluoromethoxy) phenyl] sulfonylamino] thiazole-4-carboxylic acid; 5. The compound according to any one of claims ^{1 to 4} , where each R 2 is independently selected from halogen, -CN, -C (O) NRfR ^g , where each of Rf and R ^g is H; -OCF3 and -OCH3. 6. A compound according to any one of claims 1-5, wherein each R ² is fluoro. 7. A compound according to any one of claims 1-6, wherein n is 0. 8. A compound according to any one of claims 1-7, wherein Z is a bond or is selected from —NR ¹⁰ C (O) -, —C (O) NR ¹⁰ - and —NR ¹⁰ C (O) NR ¹¹ -. 9. A compound according to any one of claims 1 to 8, wherein Z is selected from —NR ¹⁰ C (O) -, —C (O) NR ¹⁰ - and —NR ¹⁰ C (O) NR ¹¹ -. 10. A compound according to any one of claims 1-9, wherein L is a bond. 11. A compound according to any one of claims 1-10, wherein L is C _1-3 alkylene. 12. A compound according to any one of claims 1-11, wherein X is a bond. 13. A compound according to any one of claims 1-12, wherein p is 1 and R ⁷ is H or methyl. 14. A compound according to any one of claims 1-13, wherein R ⁴ is H. 15. A compound according to any one of claims 1-14, wherein R ⁵ is H and R ⁶ is H or methyl. 16. A compound according to any one of claims 1-15, where R ¹ represents H; pr represents a phenyl group; m is 2; each R ^{2 is} independently halogen; n is equal to 0; Z is selected from —NR ¹⁰ C (O) -, —C (O) NR ¹⁰ - and —NR ¹⁰ C (O) NR ¹¹ -; L is a bond or is C1 _-3 alkylene; X is a bond; p is 1; and R ⁴ is H; R ⁵ is selected from H, -CN and _C1-C2 alkyl; R ⁶ is H or methyl, R ⁷ is H or methyl. 17. A compound according to any one of claims 1-16, where R ¹ represents H; pr is phenyl; m is 2; each R ² is fluorine; n is equal to 0; Z is —NR ¹⁰ C (O) NR ¹¹ -; L is a bond; - 78 038487 X is a bond; p is 1 and R ⁷ is H; R ⁴ represents H; R ⁵ represents H; R ⁶ is H. 18. A compound according to any one of claims 1-17, wherein R ⁴ , R ⁵ , R ⁶ and R ⁷ , if present, each represent a hydrogen atom. 19. The connection according to claim 1, selected from 20. A compound which is 5 - [[3,5-difluoro-4- (guanidinocarbamoylamino) phenyl] sulfonylamino] thiazole-4-carboxylic acid or a pharmaceutically acceptable salt thereof. 21. A compound according to claim 20 which is 5 - [[3,5-difluoro-4- (guanidinocarbamoylamino) phenyl] sulfonylamino] thiazole-4-carboxylic acid. 22. A compound according to claim 20 which is 5 - [[3,5-difluoro-4 (guanidinocarbamoylamino) phenyl] sulfonylamino] thiazole-4-carboxylic acid sodium salt. 23. A pharmaceutical composition comprising a compound according to any one of claims 1-22 together with at least one pharmaceutically acceptable carrier or diluent. 24. A pharmaceutical composition according to claim 23, further comprising a β-lactam antibiotic. 25. The pharmaceutical composition of claim 24, wherein the β-lactam antibiotic is selected from carbapenems, penicillins, cephalosporins and penems. 26. The pharmaceutical composition of claim 24, wherein the β-lactam antibiotic is meropenem. 27. A pharmaceutical composition according to any one of claims 23 to 26, further comprising a serine β-lactamase inhibitor. 28. The pharmaceutical composition according to claim 27, wherein the serine-β-lactamase inhibitor is a compound of formula (II) or a pharmaceutically acceptable salt thereof [FORMULA (II)] where G is selected from —CN and —C (O) NR ^j R ^k ; R ^{k is} selected from -W and -QW; where W is selected from 5-6 membered heterocyclyl, R ^j and -N (R ^j ) 2; and Q is selected from —NR ^j C (O) -, —C (O) —NR ^j -, C1-3 alkylene, —O — C _1-3 alkylene, and —N (R ^j ) —C ₁ . ₃ αalkylene α; each R ^{j is} selected from H and unsubstituted C _1-3 alkyl, preferably H. 29. The pharmaceutical composition of claim 28, wherein the serine β-lactamase inhibitor is selected from WCK4234, avibactam, relbactam, zidebactam and nakubactam, or a pharmaceutically acceptable salt thereof, preferably WCK4234 or a pharmaceutically acceptable salt thereof. 30. Use of a compound according to any one of claims 1 to 22 for the treatment or prevention of a bacterial infection. 31. Use of a compound according to any one of claims 1 to 22 for the elimination or reduction of antibiotic resistance in gram-negative bacteria. 32. Use of a pharmaceutical composition according to any one of claims 23 to 29 for the treatment or prevention of a bacterial infection. 33. Use according to claim 30 or 32, wherein the bacterial infection is caused by bacteria selected from Enterobacteriaceae, Pseudomonas and Moraxel. 34. Use according to claim 31, wherein the bacteria are selected from Enterobacteriaceae, Pseudomonas and Moraxel. 35. Use according to claim 33 or 34, wherein the bacteria selected from Enterobacteriaceae, Pseudomonads and Moraxel are selected from Klebsiella pneumoniae, Escherichia coli, Pseudomonas aeruginosa, Burkholderia cepacia and Acinetobacter baumannii. - 80 038487 36. Use according to claim 30 or 32, wherein the bacterial infection is caused by the action of carbapenem-resistant enterobacteriaceae.