Chemical compound

Abstract

The present invention relates to a compound which is a thiazole derivative of formula (I) or a pharmaceutically acceptable salt thereof, wherein R<supgt;1< / supgt>, R<supgt;2< / supgt>, R<supgt;3< / supgt>, R<supgt;4< / supgt>, R<supgt;5< / supgt>, R<supgt;6< / supgt>, R<supgt;7< / supgt>, Z, L, X, m, n and p are as defined herein. The compound can be used for treating and preventing bacterial infections. The present invention also relates to combinations of compounds of formula (I) with additional active agents.

Description

Field of the Invention

[0001] The present invention relates to compounds belonging to thiazole derivatives. The compounds of the present invention are applied to the prevention or treatment of bacterial infections. The present invention also provides such compounds themselves and pharmaceutical compositions containing these compounds. The compounds of the present invention can be used as inhibitors of metallo-β-lactamase (MBL). The compounds of the present invention can be used in combination therapy, for example, in combination with one or more antibiotic agents and optionally with one or more serine-β-lactamase (SBL) inhibitors. Such combination therapy has particular application in the prevention or treatment of bacterial infections caused by bacteria resistant to treatment with antibiotic agents administered alone, especially when the resistance is attributed to the presence of metallo-β-lactamase and / or serine-β-lactamase and treatment with β-lactam antibiotics alone may be unsuccessful. In such cases, combination therapy can salvage the antibacterial activity of β-lactam antibiotics. Background Art

[0002] Bacteria in clinical and non-clinical settings are becoming increasingly resistant to conventional antibiotics, and this resistance is becoming a serious clinical and epidemiological problem for human health. For example, it has been shown that a single amino acid mutation in bacterial DNA-dependent RNA polymerase can reduce the binding affinity of this target enzyme for antibiotics, resulting in a high frequency of resistance (FoR). One way previously considered to address FoR was to develop a single agent that inhibits two related bacterial enzymes. Examples of such agents include gepotidacin and zoliflodacin. Gepotidacin inhibits two similar DNA processing components (GyrA and ParC) of type II and IV topoisomerases, and zoliflodacin inhibits two similar ATP hydrolysis components (GyrB and ParE) of type II and IV topoisomerases. However, this approach is not always suitable for addressing other forms of resistance, such as when microbial resistance to antibiotics arises through the production of bacterial enzymes capable of inactivating antibacterial drugs.

[0003] In Gram-negative bacteria, resistance to antibiotics, particularly β-lactam antibiotics, often arises due to the production of β-lactamases by the organism. β-lactamases include metallo-β-lactamases (MBLs) and serine-β-lactamases (SBLs). Serine β-lactamases use an active serine to hydrolyze the β-lactam ring by a covalent mechanism, while the structurally distinct metallo-β-lactamases use Zn metal coordination and a hydroxide ion to hydrolyze the β-lactam ring. In the field of bacterial β-lactamases, particularly in the Gram-negative field, and more particularly in the Enterobacteriaceae, recently evolved metallo-β-lactamases have supplemented the older serine-β-lactamases. Thus, resistance of Gram-negative bacteria to β-lactam antibiotics particularly arises due to the production of both β-lactamases by the organism.

[0004] As described above, in Gram-negative bacteria, resistance to antibiotics often arises due to the production of β-lactamases, particularly metallo-β-lactamases (MBLs), by the organism. MBLs are resistance determinants that increase clinical relevance. In fact, due to their broad spectrum, effective carbapenemase activity, and resistance to inhibitors, these enzymes can confer resistance to almost all β-lactam antibiotics.

[0005] MBLs were first discovered in the mid-1960s to be carried by mobile DNA elements in species with only low pathogenic potential. However, genes encoding MBLs spread among major Gram-negative bacteria in the 1990s, leading to a health crisis due to the international spread of carbapenem-resistant Enterobacteriaceae producing VIM-type and NDM-type metallo-β-lactamases.

[0006] Functional characteristics of these Enterobacteriaceae include effective carbapenemase activity and resistance to clinical β-lactamase inhibitors (clavulanate and sulbactam). β-Lactam resistance varies among different metallo-β-lactamases, and substrate specificity can vary from a narrow range (e.g., the CphA metallo-β-lactamase of Aeromonas hydrophila) to a broadened range (e.g., VIM-type metallo-β-lactamases, which can hydrolyze almost all classes of β-lactams except monobactams).

[0007] MBLs have three major structural subclasses, which have considerable internal diversity. Members of different subclasses differ not only in their high sequence diversity but also in the structure of their active sites. In enzymes of the B1 and B3 subclasses, the active site contains two zinc ions; in members of the B2 subclass, the active site contains only one zinc ion.

[0008] Acquired metallo-β-lactamases have been detected in strains of Enterobacteriaceae, Pseudomonas aeruginosa, Acinetobacter baumannii, and other Gram-negative bacteria. Among the acquired MBLs, almost all of the enzymes belong to subclass B1, suggesting that members of this subclass have a greater overall propensity to be captured and disseminated by mobile genetic elements than members of subclasses B2 and B3.

[0009] As examples, subclass B1 includes IMP-type, VIM-type, and NDM-type enzymes.

[0010] IMP-type enzymes include IMP-1, first discovered in Japan in the late 1980s and subsequently reported worldwide in Enterobacteriaceae and Gram-negative bacteria. IMP-type enzymes have a broad substrate specificity and high affinity for cephalosporins and carbapenems, but they have little activity against temocillin.

[0011] VIM-type enzymes include VIM-2, first discovered in Europe in the late 1990s and subsequently reported worldwide. VIM-type enzymes were initially detected in Pseudomonas aeruginosa and other Gram-negative bacteria and have since emerged in Enterobacteriaceae and have become a major problem in some settings. More than 20 different VIM isoforms are known, each with a defined geographical distribution except for VIM-1 and VIM-2, which have a wider distribution than IMP-type enzymes. VIM-type metallo-β-lactamases show a broader substrate specificity than IMP-type enzymes and are able to hydrolyze 6-α-methoxy-penicillins. In addition, VIM-type enzymes are unique among metallo-β-lactamases in that they have a high affinity for carbapenems.

[0012] New Delhi metallo-β-lactamase 1 (NDM-1) is a novel metallo-β-lactamase first discovered in a patient with Klebsiella pneumoniae infection hospitalized in New Delhi. Subsequently, Enterobacteriaceae organisms containing this new β-lactamase were found to be widely distributed in India, Pakistan, and Bangladesh and are now emerging in the United Kingdom and many other countries. New Delhi metallo-β-lactamase 1 (NDM-1) is a polypeptide 158 amino acids in length (accession number AB571289) that is able to hydrolyze a broad range of β-lactam antibiotics, including penicillins, cephalosporins, and carbapenems, which are mainstays in the treatment of antibiotic-resistant bacterial infections.

[0013] Therefore, there is an urgent need for new antibacterial compounds and compositions, as well as adjuvant therapies for treating bacterial infections, particularly those caused by bacteria expressing MBL enzymes. There is also an urgent need for new compositions for treating bacterial infections caused by bacteria showing high resistance, particularly when resistance to antibiotic agents (especially β-lactam antibiotic agents) occurs through the production by bacteria of one or more enzymes capable of inactivating the antibacterial drugs. The present invention aims to solve some or all of these problems. Summary of the Invention

[0014] Previously, the inventors reported in WO2014 / 198888 that certain thiazole derivatives are inhibitors of metallo-β-lactamases including NDM-1. The inventors have now surprisingly found that the compounds of formula (I) are effective inhibitors of metallo-β-lactamases including NDM-1 and have improved properties compared to the compounds disclosed in WO2014 / 198849. Thus, the compounds can be used for treating and preventing bacterial infections, for example, by using in combination with β-lactam antibiotics.

[0015] The inventors have also found that the compounds of formula (I) can be advantageously used in combination with inhibitors of serine-β-lactamases and other antibiotic agents such as β-lactam antibiotics (e.g., carbapenem antibiotics). Such combination therapy is particularly relevant for preventing or treating bacterial infections caused by bacteria that show high resistance to treatment by administration of antibiotic agents alone, especially when the bacterial infection is caused by bacteria producing β-lactamases.

[0016] Accordingly, the present invention provides a compound which is a thiazole derivative of formula (I) or a pharmaceutically acceptable salt thereof,

[0017]

[0018] wherein,

[0019] οR 1 is selected from H, R 1a and -CH 2 OC(O)R 1a , where R 1a is selected from unsubstituted C 1 to C 4 alkyl and phenyl;

[0020] ο is a cyclic group selected from C 6 to C 10 aryl, 5- to 10-membered heteroaryl, and 4- to 10-membered carbocyclic and heterocyclic groups;

[0021] οeach R 2 is independently selected from:

[0022] (i) Halogen or R 8 ;

[0023] (ii) C 1-3 alkyl, O(C 1-3 alkyl), S(C 1-3 alkyl), SO(C 1-3 alkyl) or SO 2 (C 1-3 alkyl), any of which may optionally be substituted by 1, 2 or 3 halogen substituents and / or one R 8 substituent; and

[0024] (iii) NR a C(O)R c and NR a C(O)NR b R c , where each R a and R b is independently selected from hydrogen and unsubstituted C 1-2 alkyl, and each R c is unsubstituted C 1-2 alkyl;

[0025] as well as

[0026] · Each R 8 is independently selected from CN, OH, -C(O)NR f R g , -NR f R g , -NR 10 C(NR 11 )R 12 , -C(NR 10 )NR 11 R 12 and -NR 10 C(NR 11 )NR 12 R 13 ; where R f and R g are each independently H or unsubstituted C 1-2 alkyl;

[0027] ο m is 0, 1, 2 or 3

[0028] ο R 3 is selected from hydrogen and C 1 to C 3 alkyl, the C 1 to C 3 alkyl being unsubstituted or substituted by a group selected from halogen, -OR 10 and -NR 10 R11 substituted by 1, 2 or 3 substituents;

[0029] n is 0 or 1

[0030] 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-, -OC(O)NR 10 -, -NR 10 C(O)S-, -SC(O)NR 10 -, -NR 10 C(NR 11 )-, -C(NR 10 )NR 11 -, -NR 10 C(NR 11 )NR 12 -, -NR 10 C(N + R 11 R 12 )-, -C(N + R 10 R 11 )NR 12 -, -NR 10 C(N + R 11 R 12 )NR 13 -, -NR 10 C(NR 11 )O-, -OC(NR 10 )NR 11 -, -NR 10 C(N + R 11 R 12 )O-, -OC(N + R 10 R 11 )NR 12 -, -NR 10 C(NR 11 )S-, -SC(NR 10 )NR 11 -, -NR 10 C(N + R 11 R 12 )S-, -SC(N + R 10 R 11 )NR 12 -, -C(O)NR15 -、-NR 10 C(O)NR 15 -、-OC(O)NR 15 、-SC(O)NR 15 、-C(NR 10 )NR 15 -、-NR 10 C(NR 11 )NR 15 -、-C(N + R 10 R 11 )NR 15 -、-NR 10 C(N + R 11 R 12 )NR 15 -、-OC(NR 10 )NR 15 、-OC(N + R 10 R 11 )NR 15 -、-SC(NR 10 )NR 15 and -SC(N + R 10 R 11 )NR 15 -;

[0031] oL is a bond or selected from C 1-4 Alkylene, C 2-4 Alkenylene, C 2-4 Alkynylidene, C 1-3 Alkylene-(C 3-6 Cycloalkylene)-C 1-3 Alkylene, C 1-4 Alkylene-(C 3-6 Cycloalkylene) and (C 3-6 Cycloalkylene)-C 1-4 Alkylene, wherein L is unsubstituted or selected from halogen, -OR 10 and-NR 10 R 11 substituted with 1 or 2 substituents; or L is -C(R 10 )=N-;

[0032] οX is a bond, or when L is not a bond or -C(R 10 )=N-, X is a bond or selected from NR 10 -、-O-、-NR 10 C(NR 11 )- and -C(NR 10 )-;

[0033] οp is 0 or 1;

[0034] οR 4 is selected from H, -CN, and C 1 to C 3 alkyl, and the C 1 to C 3 alkyl is unsubstituted or substituted with 1, 2, or 3 substituents selected from halogen, -OR 10 , -NR 10 R 11 , and -CN;

[0035] or R 4 is linked to R 5 to form together with the atoms to which they are attached a 5- to 6-membered heterocyclic group containing at least one saturated carbon atom in the ring, and the heterocyclic group is unsubstituted or substituted with 1 or 2 substituents selected from unsubstituted C 1 to C 2 alkyl, halogen, -OR 10 , -NR 10 R 11 , and -CN;

[0036] οR 5 is selected from: H, -CN, and unsubstituted or C 10 substituted with 1, 2, or 3 substituents selected from halogen, -OR 10 , -NR 11 R 1 to C 3 alkyl;

[0037] or R 5 is linked to R 4 to form together with the atoms to which they are attached a 5- to 6-membered heterocyclic group containing at least one saturated carbon atom in the ring, and the heterocyclic group is unsubstituted or substituted with 1 or 2 substituents selected from unsubstituted C 1 to C 2 alkyl, halogen, -OR 10 , -NR 10 R 11 , and -CN;

[0038] or R 5 is linked to R 6 to form together with the atoms to which they are attached a 5- to 6-membered heterocyclic group containing at least one saturated carbon atom in the ring, and the heterocyclic group is unsubstituted or substituted with 1 or 2 substituents selected from unsubstituted C 1 to C 2 alkyl, halogen, -OR 10 , -NR 10 R 11substituted by 1 or 2 substituents of -CN;

[0039] οR 6 selected from: H, -CN, and C which is unsubstituted or substituted by 1, 2 or 3 substituents selected from halogen, -OR 10 , -NR 10 R 11 and -CN; 1 to C 3 alkyl;

[0040] or R 6 is linked with R 5 to form together with the atoms to which they are attached a 5- to 6-membered heterocyclic group containing at least one saturated carbon atom in the ring, said heterocyclic group being unsubstituted or substituted by 1 or 2 substituents selected from unsubstituted C 1 to C 2 alkyl, halogen, -OR 10 , -NR 10 R 11 and -CN;

[0041] or R 6 is linked with R 7 (if present) to form together with the atoms to which they are attached a 5- to 6-membered heterocyclic group containing at least one saturated carbon atom in the ring, said heterocyclic group being unsubstituted or substituted by 1 or 2 substituents selected from unsubstituted C 1 to C 2 alkyl, halogen, -OR 10 , -NR 10 R 11 and -CN;

[0042] οR 7 (if present) selected from: H, -CN, and C which is unsubstituted or substituted by 1, 2 or 3 substituents selected from halogen, -OR 10 , -NR 10 R 11 and -CN; 1 to C 3 alkyl;

[0043] or R 7 is linked with R 6 to form together with the atoms to which they are attached a 5- to 6-membered heterocyclic group containing at least one saturated carbon atom in the ring, said heterocyclic group being unsubstituted or substituted by 1 or 2 substituents selected from unsubstituted C 1 to C 2 alkyl, halogen, -OR 10 , -NR 10 R 11 and -CN;

[0044] Each R 10 、R 11 、R 12 、R 13 and R 14 is independently H or methyl;

[0045] Each R 15 is independently a substituted C 1 to C 4 alkyl or an unsubstituted C 2 to C 4 alkyl, wherein when R 15 is a substituted alkyl, the alkyl is independently selected from halogen, CN, -OR 10 、and -NR 10 R 11 and is substituted with 1, 2 or 3 substituents.

[0046] The present invention also provides a compound of formula (I), wherein

[0047] οR 1 、 R 2 、m、R 3 、n、L、X、p、R 4 、R 5 、R 6 、R 7 (if present), R 10 、R 11 、R 12 、R 13 and R 14 are as defined herein;

[0048] οZ is a bond or is selected from -NR 10 C(O)-, -C(O)NR 10 -, -NR 10 C(O)NR 11 -, -NR 10 C(O)O-, -OC(O)NR 10 、-NR 10 C(O)S-, -SC(O)NR 10 、-NR 10 C(NR 11 )-, -C(NR 10 )NR 11 -, -NR 10 C(NR 11 )NR 12 -, -NR 10 C(N + R 11 R 12 )-, -C(N+ R 10 R 11 )NR 12 -, -NR 10 C(N + R 11 R 12 )NR 13 -, -NR 10 C(NR 11 )O-, -OC(NR 10 )NR 11 , -NR 10 C(N + R 11 R 12 )O-, -OC(N + R 10 R 11 )NR 12 -, -NR 10 C(NR 11 )S-, -SC(NR 10 )NR 11 , -NR 10 C(N + R 11 R 12 )S-, and -SC(N + R 10 R 11 )NR 12 -; and

[0049] οR 15 does not exist.

[0050] The present invention also provides a pharmaceutical composition, which comprises a compound as described herein and optionally further comprises an antibiotic agent. The pharmaceutical composition generally comprises a compound as described herein and at least one pharmaceutically acceptable carrier or diluent, and optionally further comprises (i) an antibiotic agent and / or (ii) a serine-β-lactamase inhibitor. Also provided is a product comprising a combination of a compound as described herein and an antibiotic agent.

[0051] The present invention also provides a compound as described herein for use in treating or preventing a bacterial infection in a subject in need thereof. Also provided is a method for treating or preventing a bacterial infection in a subject, which method comprises administering to the subject an effective amount of a compound as described herein. Also provided is the use of a compound as described herein in the preparation of a medicament for treating or preventing a bacterial infection in a subject.

[0052] The present invention also provides a product comprising a compound described herein, a serine-β-lactamase inhibitor, and an antibiotic agent. The product can be used to treat or prevent bacterial infections in a subject in need thereof, particularly when the bacterial infection is caused by bacteria resistant to treatment with the antibiotic agent alone, and especially when the resistance is attributable to the presence of a metallo-β-lactamase and / or a serine-β-lactamase. In patients suffering from or susceptible to infections caused by these bacteria, treatment with a β-lactam antibiotic alone may not be successful. BRIEF DESCRIPTION OF THE DRAWINGS

[0053] Figure 1 Shows the results of an in vivo efficacy study using the compound of Example 2 described herein in a mouse model. The data show that meropenem alone inhibits Klebsiella pneumoniae NTBC104 bacterial infection in mice compared to [meropenem + Example 2]. Compared to meropenem alone, 30 mg / kg of meropenem slightly reduced the bacterial load, while 30 mg / kg of meropenem plus 30 mg / kg of Example 2 showed a 1.6 Log reduction in CFU 10 , significantly reducing the bacterial load.

[0054] Figure 2 Shows the cumulative MIC - meropenem potentiation of the compounds of Example 2 and Example 26 in a panel of clinical Enterobacteriaceae strains (196 isolates) expressing the NDM enzyme. The data show that at a concentration of 8 μg / mL of Example 2 or Example 26, meropenem was potentiated to just below 90% of the strains showing a meropenem MIC of 8 μg / mL. In contrast, meropenem alone at the same concentration was only able to prevent the growth of <1% of the strains, and within the experimental parameters, meropenem alone was not able to achieve growth arrest in all 90% of the strains. DETAILED DESCRIPTION

[0055] DEFINITIONS

[0056] As used herein, C 1 to C 4 alkyl is a straight-chain or branched-chain alkyl containing 1 to 4 carbon atoms. C 1 to C 4 alkyl is generally C 1 to C 3 alkyl or C 2 to C 4 alkyl. Examples of C 1 to C 4 alkyl include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and tert-butyl. C 1 to C 3 alkyl is generally C 1 to C2 Alkyl. C 1 to C 2 The alkyl is methyl or ethyl, usually methyl. For the avoidance of doubt, when there are two alkyls, the alkyls can be the same or different.

[0057] As used herein, C 2 to C 4 The alkenyl is a straight-chain or branched-chain alkenyl having 2 to 4 carbon atoms and having one or more (e.g., one or two, usually one) double bonds. Usually, C 2 to C 4 The alkenyl is C 2 to C 3 alkenyl. C 2 to C 4 Examples of the alkenyl include vinyl, propenyl, and butenyl. For the avoidance of doubt, when there are two alkenyls, the alkenyls can be the same or different.

[0058] As used herein, C 2 to C 4 The alkynyl is a straight-chain or branched-chain alkynyl having 2 to 4 carbon atoms and having one or more (e.g., one or two, usually one) triple bonds. Usually, C 2 to C 4 The alkynyl is C 2 to C 3 alkynyl. C 2 to C 4 Examples of the alkynyl include ethynyl, propynyl, and butynyl. For the avoidance of doubt, when there are two alkynyls, the alkynyls can be the same or different.

[0059] As used herein, C 1 to C 4 The alkylene is an unsubstituted or substituted bidentate moiety obtained by removing two hydrogen atoms from a C 1 to C 4 alkane. The two hydrogen atoms can be removed from the same carbon atom or different carbon atoms. Usually, C 1 to C 4 The alkylene is C 1 to C 3 alkylene. C 1 to C 4 Examples of the alkylene include methylene, ethylene, n-propylene, isopropylidene, n-butylene, sec-butylene, and tert-butylene. C 1 to C 4 The alkylene is usually C 1 to C 2 alkylene. C 1 to C 2The alkyl group is methylene or ethylene group, usually methylene group. To avoid ambiguity, when there are two alkylene groups, the alkylene groups can be the same or different.

[0060] As used herein, C 2 to C 4 The alkenylene group is an unsubstituted or substituted bidentate moiety obtained by removing two hydrogen atoms from an alkene having from C 2 to C 4 The two hydrogen atoms can be removed from the same carbon atom or different carbon atoms. Generally, the alkenylene group having from C 2 to C 4 is an alkenylene group having from C 2 to C 3 Examples of the alkenylene group having from C 2 to C 4 include vinylidene group, n-propenylene group, isopropenylene group, n-butenylene group, sec-butenylene group and tert-butenylene group. The alkenylene group having from C 2 to C 3 is generally an alkenylene group having from C 2 i.e., vinylidene group. To avoid ambiguity, when there are two alkenylene groups, the alkenylene groups can be the same or different.

[0061] As used herein, C 2 to C 4 The alkynylene group is an unsubstituted or substituted bidentate moiety obtained by removing two hydrogen atoms from an alkyne having from C 2 to C 4 The two hydrogen atoms can be removed from the same carbon atom or different carbon atoms. Generally, the alkynylene group having from C 2 to C 4 is an alkynylene group having from C 2 to C 3 Examples of the alkynylene group having from C 2 to C 4 include ethynylene group, n-propynylene group, isopropynylene group, n-butynylene group, sec-butynylene group and tert-butynylene group. The alkynylene group having from C 2 to C 3 is generally an alkynylene group having from C 2 i.e., ethynylene group. To avoid ambiguity, when there are two alkynylene groups, the alkynylene groups can be the same or different.

[0062] The alkyl group, alkenyl group, alkynyl group, alkylene group, alkenylene group or alkynylene group as used herein can be unsubstituted or substituted. Unless otherwise specified, the substituted alkyl group, alkenyl group or alkynyl group usually bears one or more, such as 1, 2, 3 or 4, such as 1, 2 or 3, such as 1 or 2, such as 1 substituents selected from -CN, -R 8 , -OR 10 and -NR 10 R 11 wherein R10 and R 11 As defined herein. Unless otherwise specified, the substituents on a substituted alkyl, alkenyl or alkynyl are generally unsubstituted themselves. When there are more than one substituent, they can be the same or different.

[0063] As used herein, halogen is generally chlorine, fluorine, bromine or iodine, preferably chlorine, bromine or fluorine, especially chlorine or fluorine, especially fluorine.

[0064] A 3- to 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 generally saturated. A 3- to 10-membered partially unsaturated carbocyclic group is a cyclic hydrocarbon containing 3 to 10 carbon atoms and containing 1 or 2 (e.g., 1) double bonds. A 3- to 10-membered carbocyclic group is generally a 4- to 10-membered carbocyclic group. Generally, a 3- to 10-membered carbocyclic group is a 3- to 6-membered carbocyclic group, such as a 4- to 6-membered or 5- to 6-membered carbocyclic group. A 3- to 10-membered carbocyclic group can be a fused bicyclic group as defined herein. A 3- to 10-membered carbocyclic group can be a saturated 4- to 6-membered, preferably 5- or 6-membered carbocyclic group. Examples of 3- to 6-membered saturated carbocyclic groups include cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

[0065] A 3- to 10-membered heterocyclic group is a cyclic group containing 3 to 10 atoms selected from C, O, N and S in the ring, the cyclic group containing at least one heteroatom, generally containing one or two heteroatoms. One or more heteroatoms are generally selected from O, N and S, most usually from S and N, especially N. For example, when the heterocyclic group represents a nitrogen-containing heterocyclic group, it contains one nitrogen atom and optionally contains additional heteroatoms selected from O, N and S. The heterocyclic group can be saturated or partially unsaturated. A 3- to 10-membered partially unsaturated heterocyclic group is a cyclic group containing 3 to 10 atoms selected from C, O, N and S and containing 1 or 2 (e.g., 1) double bonds.

[0066] A 3- to 10-membered heterocyclic group is generally a 4- to 10-membered heterocyclic group. Sometimes, a 3- to 10-membered heterocyclic group is a 3- to 6-membered heterocyclic group, such as a monocyclic 4- to 6-membered heterocyclic group or a monocyclic 5- or 6-membered heterocyclic group. Alternatively, a 3- to 10-membered heterocyclic group can be a 9- or 10-membered fused bicyclic heterocyclic group (i.e., a fused heterobicyclic group).

[0067] Examples of 5- and 6-membered saturated heterocyclic groups as defined herein include piperazine, piperidine, morpholine, 1,3-oxazolidine, pyrrolidine, imidazolidine and oxazolidine, including their quaternized derivatives. Examples of 5- and 6-membered partially saturated heterocyclic groups as defined herein include tetrahydropyrazine, tetrahydropyridine, dihydro-1,4-oxazine, tetrahydropyrimidine, dihydro-1,3-oxazine, dihydropyrrole, dihydroimidazole and dihydrooxazole, and include their quaternized derivatives.

[0068] As defined herein, examples of 9- and 10-membered fused heterobicyclic groups include: 9-membered fused heterobicyclic groups such as indoline, 2,3-dihydrobenzofuran, 2,3-dihydrobenzothiophene, 2,3-dihydro-1H-benzimidazole, 2,3-dihydrobenzoxazole, 2,3-dihydrobenzothiazole, benzodioxole, 4,5,6,7-tetrahydrothiazolo[5,4-c]pyridine and 4,5,6,7-tetrahydrothiazolo[4,5-c]pyridine, including their quaternized derivatives; and 10-membered heterobicyclic groups such as 1,2,3,4-tetrahydroquinoline, 1,2,3,4-tetrahydroisoquinoline, chroman, isochroman, dihydrobenzothiopyran, isothiochromane, 1,2,3,4-tetrahydroquinoxaline, 1,2,3,4-tetrahydroquinazoline, 1,4-dihydro-2H-benzo[d][1,3]oxazine, 3,4-dihydro-2H-benzo[b][1,4]oxazine, 3,4-dihydro-2H-benzo[b][1,4]thiazine, 1,4-dihydro-2H-benzo[d][1,3]thiazine, 4H-benzo[d][1,3]dioxin and 2,3-dihydrobenzo[b][1,4]dioxin, and including their quaternized derivatives. Generally, the fused heterobicyclic group contains 1, 2 or 3, preferably 1 or 2 nitrogen atoms.

[0069] For the avoidance of doubt, reference to a heterocyclic group also includes fused polycyclic systems, including for example fused bicyclic systems in which a heterocyclic group is fused to an aryl group. When the heterocyclic group is such a fused heterocyclic group, preferred examples are fused ring systems in which a 5- to 6-membered heterocyclic group is fused to a phenyl group.

[0070] As used herein, C 3 to C 6 subcycloalkyl (also referred to as C 3-6 subcycloalkyl) is an unsubstituted or substituted bidentate moiety obtained by removing two hydrogen atoms from a saturated C 3 to C 6 carbocyclic group. The two hydrogen atoms can be removed from the same carbon atom or different carbon atoms. Examples of C 3-6 subcycloalkyl include cyclopropylidene, cyclobutylidene, cyclopentylidene and cyclohexylidene.

[0071] As used herein, C 6 to C 10 aryl is a substituted or unsubstituted monocyclic or fused polycyclic aromatic group containing 6 to 10 carbon atoms in the ring portion. Examples include monocyclic groups (e.g., phenyl) and fused bicyclic groups (e.g., naphthyl and indenyl). Phenyl (benzene) is preferred.

[0072] As used herein, a 5- to 10-membered heteroaryl is a substituted or unsubstituted monocyclic or fused polycyclic aromatic group containing 5 to 10 atoms in the ring portion and including at least one heteroatom usually selected from O, S, and N, such as 1, 2, or 3 heteroatoms. The heteroaryl is usually a 5- or 6-membered heteroaryl or a 9- or 10-membered heteroaryl, preferably a 5- or 6-membered heteroaryl. Preferably, the heteroaryl contains 1, 2, or 3, preferably 1 or 2 nitrogen atoms.

[0073] Examples of 5- and 6-membered heteroaryls include pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyridine, pyridazine, pyrimidine, and pyrazine. Examples of 9- and 10-membered heteroaryls include: 9-membered heteroaryls such as indole, benzothiophene, benzofuran, benzoxazole, benzothiazole, benzimidazole, imidazo[1,2-a]pyridine, [1,2,4]triazolo[1,5-a]pyridine, and imidazo[1,2-a]pyrazine, including their quaternized derivatives; and 10-membered heteroaryls such as quinoline, isoquinoline, quinazoline, and quinoxaline.

[0074] To avoid doubt, the mention of heteroaryl also includes fused polycyclic systems, including for example fused bicyclic systems in which a heteroaryl is fused to an aryl. When the heteroaryl is such a fused heteroaryl, preferred examples are fused ring systems in which a 5- to 6-membered heteroaryl is fused to a phenyl group.

[0075] As used herein, a fused bicyclic group is a group containing two cyclic moieties that share a common bond between two atoms.

[0076] As described herein, a carbocyclic group, a heterocyclic group, an aryl, or a heteroaryl can be unsubstituted or substituted. For example, a carbocyclic group, a heterocyclic group, an aryl, or a heteroaryl can be unsubstituted or substituted by 1, 2, or 3, usually 1 or 2, such as 1 substituent. Suitable substituents include halogen; -CN; OR 10 and -NR 10 R 11 (wherein R 10 and R 11 are as defined herein) unsubstituted C 1 -C 2 alkyl and R as shown in formula (I) and defined herein 2 . Unless otherwise specified, the substituents on a substituted carbocyclic group, heterocyclic group, aryl, or heteroaryl are usually unsubstituted themselves.

[0077] The compounds of the present invention may contain a heterocyclic group or heteroaryl containing at least one nitrogen atom. In these compounds, the nitrogen atoms are independently selected from secondary, tertiary and quaternary nitrogen atoms. 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 is formed by bonding an additional alkyl group to a nitrogen atom in the moiety such that the valence of the nitrogen atom increases from 3 to 4 and the nitrogen atom becomes positively charged.

[0078] As used herein, a pharmaceutically acceptable salt is a salt of a pharmaceutically acceptable acid or base. Pharmaceutically acceptable acids include inorganic acids and organic acids, such as hydrochloric acid, sulfuric acid, phosphoric acid, diphosphoric acid, hydrobromic acid or nitric acid, and organic acids such as oxalic acid, citric acid, fumaric acid, maleic acid, malic acid, ascorbic acid, succinic acid, tartaric acid, benzoic acid, acetic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid or p-toluenesulfonic acid. Pharmaceutically acceptable bases include alkali metals (such as sodium or potassium) and alkaline earth metals (such as calcium or magnesium) hydroxides and organic bases, such as alkylamines, aralkylamines and heterocyclic amines. The hydrochloride and acetate salts are preferred, especially the hydrochloride salt.

[0079] In formula (I), the stereochemistry is not restricted. In particular, compounds of formula (I) containing one or more chiral centers can be used in enantiomerically pure or diastereomerically pure form, or in the form of a mixture of isomers. In addition, to avoid doubt, the compounds of the present invention can be used in any tautomeric form. Generally, the medicaments or substances described herein contain at least 50%, preferably at least 60%, 75%, 90% or 95% of enantiomerically pure or diastereomerically pure compounds according to formula (I). Generally, the compounds of the present invention contain at least 60%, such as at least 75%, 90% or 95% of a single enantiomer or diastereomer. Preferably, the compound is substantially optically pure.

[0080] To avoid doubt, the terms "thiazole derivative" and "thiazolyl derivative" are used interchangeably and refer to the compounds of the present invention, such as compounds of formula (I), unless otherwise stated.

[0081] The compounds of the present invention

[0082] Typically, in formula (I), R 1 is selected from H and R 1a . More preferably, R 1 is H. R 1a is generally unsubstituted C 1 to C 4 alkyl, such as unsubstituted C 1 to C 2 alkyl. More preferably, R 1a is methyl or tert-butyl.

[0083] In formula (I), may preferably be a cyclic group selected from C 6 to C 10 aryl and 5- to 10-membered heteroaryl. Preferably a cyclic group selected from phenyl, 5- to 6-membered heteroaryl, and 5- to 6-membered carbocyclic and heterocyclic groups. More preferably selected from phenyl and 5- to 6-membered heteroaryl. Even more preferably, is phenyl.

[0084] When is a 5- to 10-membered heteroaryl, it is preferably a 5- or 6-membered group. When is a 4- to 10-membered heterocyclic or carbocyclic group, it is preferably a 5- or 6-membered group. When is a heterocyclic group or heteroaryl, it preferably contains 1, 2 or 3, preferably 1 or 2 heteroatoms selected from O, N and S. When is a heterocyclic group or heteroaryl, it is preferably a nitrogen-containing group. When is a fused heteroaryl or heterocyclic group, preferably contains a benzene ring fused to a 5- or 6-membered heterocyclic group or heteroaryl as defined herein.

[0085] Preferably, selected from phenyl, cyclohexane, piperidine, pyridazine, pyridine and thiazole. More preferably, selected from phenyl, pyridazine, pyridine and thiazole. Even more preferably, is phenyl.

[0086] In formula (I), each R 2 is independently selected from:

[0087] (i) halogen or R 8 ;

[0088] (ii) C 1-3 alkyl, O(C 1-3 alkyl), S(C 1-3 alkyl), SO(C 1-3 alkyl) or SO 2 (C 1-3 alkyl), where any one may optionally be substituted by 1, 2 or 3 halogen substituents and / or one R 8 substituent; and

[0089] (iii) NR a C(O)R c and NR a C(O)NR b R c where each R aand R b are independently selected from hydrogen and unsubstituted C 1-2 alkyl, and each R c is unsubstituted C 1-2 alkyl;

[0090] wherein R 8 is as defined herein.

[0091] When the R 2 group is according to option (i) above, preferably the group is a halogen group. Fluorine is preferred.

[0092] When the R 2 group is according to option (ii) above, preferably, the C 2 in the R 1-3 alkyl is C 1-2 alkyl, more preferably C 1 alkyl (methyl). The R 2 group is preferably selected from C 1-2 alkyl, O(C 1-2 alkyl), S(C 1-2 alkyl) and SO(C 1-2 alkyl); more preferably C 1-2 alkyl and O(C 1-2 alkyl), wherein as described above, each may be unsubstituted or substituted. When the R 2 group is according to option (ii) above, the R 2 group may optionally be substituted with 1, 2 or 3 halogen substituents and / or one R 8 substituent; preferably substituted with 1, 2 or 3 halogen substituents (one or more, preferably all fluorine) or one R 8 substituent. Preferably, the R 2 group according to option (ii) includes C 1-2 alkyl and O(C 1-2 alkyl) which are each unsubstituted or substituted with 3 fluorine substituents (e.g., -CF 3 , -OCF 3 and -OCH 3 ). To avoid doubt, when R 2 is according to option (ii) above and is substituted as described above, one or more substituents are each preferably present on the alkyl portion of the R 2 group.

[0093] When the R 2 group is according to option (iii) above, each R a and R b are independently preferably selected from hydrogen and methyl. Each R c is preferably methyl. More preferably, each R a and Rb independently selected from hydrogen and methyl (preferably hydrogen) and R c is methyl.

[0094] Preferably, each R 8 group is independently selected from CN, OH, -C(O)NR f R g , -NR f R g , where R f and R g are each independently H or methyl, preferably hydrogen. More preferably, each R 8 group is independently selected from CN and -C(O)NR f R g .

[0095] Thus, in formula (I), each R 2 is preferably independently selected from:

[0096] · halogen or R 8 ;

[0097] · C 1-2 alkyl, O(C 1-2 alkyl), S(C 1-2 alkyl), SO(C 1-2 alkyl) or SO 2 (C 1-2 alkyl), where any one of them can optionally be substituted by 1, 2 or 3 halogen substituents and / or one R 8 substituent; and

[0098] · NR a C(O)R c and NR a C(O)NR b R c , where each R a and R b are independently selected from hydrogen and unsubstituted C 1-2 alkyl, and each R c is unsubstituted C 1-2 alkyl;

[0099] where each R 8 is independently selected from CN, OH, -C(O)NR f R g and -NR f R g ; where R f and R g are each independently H or unsubstituted C 1-2 alkyl.

[0100] More preferably, in formula (I), each R 2 is independently selected from

[0101] · halogen, CN, OH, -C(O)NR f R g 、-NR f R g ; wherein R f and R g are each independently H or methyl; and

[0102] · C 1-2 alkyl, O(C 1-2 alkyl), S(C 1-2 alkyl), SO(C 1-2 alkyl), wherein any one of them may optionally be substituted with 1, 2 or 3 halogen substituents and / or one substituent selected from CN and OH.

[0103] In formula (I), m is preferably 0, 1 or 2. More preferably, m is 1 or 2. Sometimes m is 1. Sometimes m is 2.

[0104] Therefore, in formula (I), it is preferred that:

[0105] · is a cyclic group selected from phenyl, 5- to 6-membered heteroaryl, and 5- to 6-membered carbocyclic and heterocyclic groups

[0106] · Each R 2 is independently selected from:

[0107] ο halogen or R 8 ;

[0108] ο C 1-2 alkyl, O(C 1-2 alkyl), S(C 1-2 alkyl), SO(C 1-2 alkyl) or SO 2 (C 1-2 alkyl), wherein any one of them may optionally be substituted with 1, 2 or 3 halogen substituents and / or one R 8 substituent; and

[0109] ο NR a C(O)R c and NR a C(O)NR b R c wherein each R a and R b are independently selected from hydrogen and unsubstituted C 1-2 alkyl, and each R c is unsubstituted C 1-2 alkyl;

[0110] · Each R 8 is independently selected from CN, OH, -C(O)NR f R g and -NR f R g ; wherein R f and R g are each independently H or unsubstituted C 1-2 alkyl.

[0111] · m is 0, 1 or 2.

[0112] More preferably, in formula (I):

[0113] · is selected from phenyl, cyclohexane, piperidine, pyridazine, pyridine and thiazole;

[0114] · Each R 2 is independently selected from

[0115] ο halogen, CN, OH, -C(O)NR f R g , -NR f R g ; wherein R f and R g are each independently H or methyl; and

[0116] ο C 1-2 alkyl, O(C 1-2 alkyl), S(C 1-2 alkyl), SO(C 1-2 alkyl), where any one of them may optionally be substituted with 1, 2 or 3 halogen substituents and / or one substituent selected from CN and OH;

[0117] And

[0118] · m is 1 or 2.

[0119] Generally, in formula (I), n is 0.

[0120] In formula (I), if n is 1, then R 3 is preferably selected from hydrogen and unsubstituted C 1 to C 3 alkyl, such as methyl or ethyl, preferably methyl. More preferably, R 3 (if present) is hydrogen.

[0121] Generally, in formula (I), Z is a bond or selected from -NR 10 C(O)-, -C(O)NR 10 -, -NR 10 C(O)NR11 -, -NR 10 C(O)O-, -OC(O)NR 10 -, -NR 10 C(O)S-, -SC(O)NR 10 -, -NR 10 C(NR 11 )-, -C(NR 10 )NR 11 -, -NR 10 C(NR 11 )NR 12 -,-NR 10 C(NR 11 )O-, -OC(NR 10 )NR 11 ,

[0122] -NR 10 C(NR 11 )S-, -SC(NR 10 )NR 11 , -C(O)NR 15 -, -NR 10 C(O)NR 15 -, -OC(O)NR 15 , -SC(O)NR 15 , -C(NR 10 )NR 15 -, -NR 10 C(NR 11 )NR 15 , -OC(NR 10 )NR 15 and -SC(NR 10 )NR 1 , wherein R 10 , R 11 , R 12 and R 15 is as defined herein. Preferably, 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-, -OC(O)NR 10 , -NR 10 C(O)S-, -SC(O)NR 10 , -NR 10 C(NR 11 )-, -C(NR 10 )NR 11 -, -NR 10 C(NR11 )NR 12 -,-NR 10 C(NR 11 )O-、-OC(NR 10 )NR 11 、-NR 10 C(NR 11 )S- and -SC(NR 10 )NR 11 , where R 10 , R 11 and R 12 As defined herein. More preferably, 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-、-OC(O)NR 10 、-NR 10 C(O)S-、-SC(O)NR 10 、-NR 10 C(NR 11 )-、-C(NR 10 )NR 11 -and-NR 10 C(NR 11 )NR 12 -. Still more preferably, 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-、-NR 10 C(O)S- and -NR 10 C(NR 11 Most preferably, Z is selected from -NR 10 C(O)-、-C(O)NR 10 -and-NR 10 C(O)NR 11 -, preferably -NR 10 C(O)-.

[0123] Typically, each R 15 Independently substituted C 1 To C 3 Alkyl or unsubstituted C 2 To C 3 Alkyl, more preferably, each R 15 independently substituted C 1 To C 2alkyl or unsubstituted C 2 alkyl; more preferably, each R 15 is independently a substituted or unsubstituted C 2 alkyl. When R 15 is a substituted alkyl, the alkyl is usually substituted by 1, 2 or 3, preferably 1 or 2, more preferably 1 substituent independently selected from halogen, CN and OR 10 , more preferably selected from CN and -OR 10 , and most preferably selected from CN and OH.

[0124] In formula (I), L is a bond or is selected from C 1-4 alkylene, C 2-4 alkenylene, C 2-4 alkynylene, C 1-3 alkylene-(C 3-6 cycloalkylene)-C 1-3 alkylene, C 1-4 alkylene-(C 3-6 cycloalkylene) and (C 3-6 cycloalkylene)-C 1-4 alkylene, where L is unsubstituted or substituted by 1 or 2 substituents selected from halogen, -OR 10 and -NR 10 R 11 ; or L is -C(R 10 )=N-. Generally, in formula (I), L is unsubstituted or substituted by 1 substituent selected from halogen, -OR 10 and -NR 10 R 11 ; most typically L is unsubstituted. When L is not a bond or -C(R 10 )=N- and L is substituted by one or more substituents as described above, the one or more substituents are each preferably present on the alkylene, alkenylene or alkynylene of L. For the avoidance of doubt, when L is a bond, L is unsubstituted.

[0125] L is preferably a bond or is selected from C 1-4 alkylene, C 2-4 alkenylene and C 2-4 alkynylene; or L is -C(R 10 )=N-. More preferably, L is a bond or is selected from C 1-3 alkylene and C 2-3 alkenylene, or is -C(R 10 )=N-. Even more preferably, L is selected from C 1-3 alkylene and C 2-3 alkenylene.

[0126] Generally, in formula (I), X is a bond, or when L is not a bond or -C(R 10) = N-, X is a bond or selected from -NR 10 -, and -O-. More preferably, X is a bond.

[0127] Therefore, preferably, in formula (I):

[0128] · 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-, -OC(O)NR 10 , -NR 10 C(O)S-, -SC(O)NR 10 , -NR 10 C(NR 11 ), -C(NR 10 )NR 11 -, and -NR 10 C(NR 11 )NR 12 ;

[0129] · L is a bond or selected from C 1-4 alkylene, C 2-4 alkenylene, and C 2-4 alkynylene; or L is -C(R 10 ) = N-;

[0130] And

[0131] · X is a bond.

[0132] More preferably, in formula (I):

[0133] · Z is selected from -NR 10 C(O)-, -C(O)NR 10 -, and -NR 10 C(O)NR 11 -;

[0134] · L is selected from C 1-3 alkylene and C 2-3 alkenylene, each of which is preferably unsubstituted;

[0135] And

[0136] · X is a bond.

[0137] In formula (I), R 4 :

[0138] (i) is selected from H, -CN, and unsubstituted or substituted by a group selected from halogen, -OR 10 (ii), -NR 10 R11 C substituted with 1, 2 or 3 substituents of -CN 1 to C 3 alkyl;

[0139] or

[0140] (ii) joined together with R 5 to form, together with the atoms to which they are attached, a 5- to 6-membered heterocyclic group containing at least one saturated carbon atom in the ring, said heterocyclic group being unsubstituted or substituted with 1 or 2 substituents selected from unsubstituted C 1 to C 2 alkyl, halogen, -OR 10 , -NR 10 R 11 and -CN.

[0141] In formula (I), R 5 :

[0142] (i) selected from H, -CN, and C 10 substituted with 1, 2 or 3 substituents selected from halogen, -OR 10 R 11 and -CN, from C 1 to C 3 alkyl;

[0143] or

[0144] (ii) joined together with R 4 to form, together with the atoms to which they are attached, a 5- to 6-membered heterocyclic group containing at least one saturated carbon atom in the ring, said heterocyclic group being unsubstituted or substituted with 1 or 2 substituents selected from unsubstituted C 1 to C 2 alkyl, halogen, -OR 10 , -NR 10 R 11 and -CN;

[0145] or

[0146] (iii) joined together with R 6 to form, together with the atoms to which they are attached, a 5- to 6-membered heterocyclic group containing at least one saturated carbon atom in the ring, said heterocyclic group being unsubstituted or substituted with 1 or 2 substituents selected from unsubstituted C 1 to C 2 alkyl, halogen, -OR 10 , -NR 10 R 11 and -CN.

[0147] When R 4 is C according to option (i) above1 to C 3 alkyl group, it is usually unsubstituted or substituted by 1, 2 or 3 halogen substituents or by 1 or 2 halogen substituents and / or substituted by a substituent selected from -OR 10 , -NR 10 R 11 and -CN. When R 4 is according to the above option (i), R 4 is preferably: H, or unsubstituted or substituted by 1, 2 or 3 halogen substituents or a -OR 10 substituent of C 1 to C 2 alkyl; more preferably R 4 is H or methyl, most preferably H.

[0148] When R 5 is C 1 to C 3 alkyl according to the above option (i), it is usually unsubstituted, or substituted by 1, 2 or 3 halogen substituents or by 1 or 2 halogen substituents and / or substituted by a substituent selected from -OR 10 , -NR 10 R 11 and -CN. When R 5 is according to the above option (i), R 5 is preferably selected from: H, -CN, and unsubstituted or substituted by 1, 2 or 3 halogen substituents or a -NR 10 R 11 substituent of C 1 to C 2 alkyl; more preferably, R 5 is H or methyl, most preferably H.

[0149] When R 4 is according to the above option (ii) and R 5 is according to the above option (ii), R 4 and R 5 are linked together to form, together with the atoms to which they are attached, a 5- to 6-membered heterocyclic group containing at least one saturated carbon atom in the ring, and the heterocyclic group is preferably unsubstituted or substituted by 1 substituent selected from unsubstituted C 1 to C 2 alkyl, halogen and -OR 10 ; more preferably, the heterocyclic group is unsubstituted or substituted by 1 substituent selected from methyl and methoxy; most preferably, the heterocyclic group is unsubstituted. Preferably, when R 4 is according to the above option (ii) and R 5 is according to the above option (ii), R 4 and R5 Joined together to form, together with the atoms to which they are attached, a 5-membered heterocyclic group, preferably 4,5-dihydro-1H-imidazole.

[0150] In formula (I), R 6

[0151] (i) is selected from: H, -CN, and C 10 to C 10 alkyl which is unsubstituted or substituted by 1, 2 or 3 substituents selected from halogen, -OR 11 and -CN; 1 to C 3 alkyl;

[0152] or

[0153] (ii) is joined to R 5 to form, together with the atoms to which they are attached, a 5- to 6-membered heterocyclic group containing at least one saturated carbon atom in the ring, said heterocyclic group being unsubstituted or substituted by 1 or 2 substituents selected from unsubstituted C 1 to C 2 alkyl, halogen, -OR 10 and -CN; 10 R 11 and -CN;

[0154] or

[0155] (iii) is joined to R 7 (if present) to form, together with the atoms to which they are attached, a 5- to 6-membered heterocyclic group containing at least one saturated carbon atom in the ring, said heterocyclic group being unsubstituted or substituted by 1 or 2 substituents selected from unsubstituted C 1 to C 2 alkyl, halogen, -OR 10 and -CN; 10 R 11 and -CN.

[0156] When R 6 is C 1 to C 3 alkyl according to option (i) above, it is usually unsubstituted or substituted by 1, 2 or 3 halogen substituents or by 1 or 2 halogen substituents and / or by one substituent selected from -OR 10 and -CN. When R 10 R 11 is according to option (i) above, R 6 is preferably selected from: H, -CN, and unsubstituted or substituted by 1, 2 or 3 halogen substituents or one -NR 6 R 10 R11 Substituted C 1 to C 2 alkyl; more preferably, R 6 is H or methyl, most preferably H.

[0157] When R 5 is according to option (iii) above and R 6 is according to option (ii) above, R 6 and R 6 are linked together to form, together with the atoms to which they are attached, a 5- to 6-membered heterocyclic group containing at least one saturated carbon atom in the ring, which heterocyclic group is preferably unsubstituted or substituted by 1 substituent selected from unsubstituted C 1 to C 2 alkyl, halogen, and -OR 10 ; more preferably, the heterocyclic group is unsubstituted or substituted by 1 substituent selected from methyl and methoxy; most preferably, the heterocyclic group is unsubstituted. Preferably, when R 5 is according to option (iii) above and R 6 is according to option (ii) above, R 5 and R 6 are linked together to form, together with the atoms to which they are attached, a 6-membered heterocyclic group, preferably morpholine or piperazine, more preferably morpholine.

[0158] In formula (I), p is 0 or 1.

[0159] In formula (I), R 7 (if present)

[0160] (i) is selected from H, -CN, and unsubstituted or C 10 substituted by 1, 2, or 3 substituents selected from halogen, -OR 10 R 11 and -CN 1 to C 3 alkyl;

[0161] or

[0162] (ii) is linked together with R 6 to form, together with the atoms to which they are attached, a 5- to 6-membered heterocyclic group containing at least one saturated carbon atom in the ring, said heterocyclic group being unsubstituted or substituted by 1 or 2 substituents selected from unsubstituted C 1 to C 2 alkyl, halogen, -OR 10 , -NR 10 R 11 and -CN.

[0163] When R 7is present and is C according to option (i) above 1 -C 3 When it is alkyl, it is usually unsubstituted or substituted with 1, 2 or 3 halogen substituents or substituted with 1 or 2 halogen substituents and / or substituted with a substituent selected from -OR 10 , -NR 10 R 11 and -CN. When R 7 is present and according to option (i) above, R 7 is preferably selected from: H, -CN, and unsubstituted or substituted with 1, 2 or 3 halogen substituents or a -NR 10 R 11 substituent-substituted C 1 to C 2 alkyl; more preferably, R 7 if present, is H or methyl, most preferably H.

[0164] When R 7 is present and R 6 is according to option (iii) above and R 7 is according to option (ii) above such that R 6 and R 7 are linked together to form, together with the atoms to which they are attached, a 5- to 6-membered heterocyclic group containing at least one saturated carbon atom in the ring, the heterocyclic group is preferably unsubstituted or substituted with 1 substituent selected from unsubstituted C 1 to C 2 alkyl, halogen and -OR 10 ; more preferably, the heterocyclic group is unsubstituted or substituted with 1 substituent selected from methyl and methoxy; most preferably, the heterocyclic group is unsubstituted. Preferably, when R 7 is present and is according to option (ii) above and R 6 is according to option (iii) above, R 6 and R 7 are linked together to form, together with the atoms to which they are attached, a 5-membered heterocyclic group, preferably imidazolidine.

[0165] Thus, preferably, in formula (I), p is 1, and R 7 is H or methyl, or is linked together with R 6 to form, together with the atoms to which they are attached, an unsubstituted 5- to 6-membered heterocyclic group. Preferably, R 4 is H or is linked together with R 5 to form, together with the atoms to which they are attached, an unsubstituted 5- to 6-membered heterocyclic group. More preferably, in formula (I), R 5 is selected from: H, -CN, and unsubstituted or substituted with 1, 2 or 3 halogen substituents or a -NR10 R 11 C substituted by substituents 1 to C 2 alkyl; and R 6 is H or methyl. Most preferably, R 4 、R 5 、R 6 and R 7 (if present) are each independently selected from methyl and hydrogen, preferably hydrogen.

[0166] For the avoidance of doubt, a heterocyclic group containing at least one saturated carbon atom in the ring contains an -CH 2 - group within the ring, where one or both hydrogen atoms of the -CH 2 - group may be substituted as defined herein. Generally, the saturated carbon atoms in the ring are unsubstituted; that is, a heterocyclic group containing at least one saturated carbon atom in the ring generally contains an -CH 2 - group within the ring. Thus, a heterocyclic group containing at least one saturated carbon atom in the ring is saturated or partially saturated. A heterocyclic group containing at least one saturated carbon atom in the ring is not aromatic.

[0167] Thus, in some preferred compounds of formula (I),

[0168] ·R 1 is H;

[0169] · is a cyclic group selected from phenyl, 5- to 6-membered heteroaryl, and 5- to 6-membered carbocyclic and heterocyclic groups;

[0170] ·m is 0, 1 or 2;

[0171] ·Each R 2 is independently selected from:

[0172] ο halogen or R 8 ;

[0173] ο C 1-3 alkyl, O(C 1-3 alkyl), S(C 1-3 alkyl), SO(C 1-3 alkyl) or SO 2 (C 1-3 alkyl), where any one of them may optionally be substituted by 1, 2 or 3 halogen substituents and / or one R 8 substituent; and

[0174] ο NR a C(O)R c and NR a C(O)NR b R c, wherein each R a and R b is independently selected from hydrogen and unsubstituted C 1-2 alkyl, and each R c is unsubstituted C 1-2 alkyl;

[0175] · Each R 8 is independently selected from CN, OH; -C(O)NR f R g , -NR f R g , wherein R f and R g are each independently H or unsubstituted C 1-2 alkyl;

[0176] · n is 0; or n is 1 and R 3 is H.

[0177] · Z is selected from -NR 10 C(O)-, -C(O)NR 10 -, -NR 10 C(O)NR 11 -, -NR 10 C(O)O-, -OC(O)NR 10 , -NR 10 C(O)S-, -SC(O)NR 10 , -NR 10 C(NR 11 )-, -C(NR 10 )NR 11 -, and -NR 10 C(NR 11 )NR 12 -;

[0178] · L is a bond or is selected from C 1-4 alkylene, C 2-4 alkenylene, and C 2-4 alkynylene; or L is -C(R 10 )=N-;

[0179] · X is a bond;

[0180] · i) p is 0;

[0181] R 4 is H; and R 5 is selected from: H, -CN, and unsubstituted or substituted with 1, 2, or 3 halogen substituents or substituted with a -NR 10 R 11 substituent of C 1 to C 2 alkyl; or R4 linked to R 5 to form, together with the atoms to which they are linked, an unsubstituted 5- or 6-membered heterocyclic group; and

[0182] R 6 is H or methyl;

[0183] or

[0184] ii) p is 1; and

[0185] R 4 is H; R 5 is selected from H, -CN, and unsubstituted or substituted by 1, 2 or 3 halogen substituents or a -NR 10 R 11 substituent substituted C 1 to C 2 alkyl; R 6 is H or methyl, R 7 is H or methyl; or R 4 linked to R 5 to form, together with the atoms to which they are linked, an unsubstituted 5- or 6-membered heterocyclic group; R 6 is H or methyl, R 7 is H;

[0186] In some even more preferred compounds of formula (I),

[0187] · R 1 is H;

[0188] · is phenyl, cyclohexane, piperidine, pyridazine, pyridine and thiazole;

[0189] · m is 1 or 2;

[0190] · Each R 2 is independently selected from:

[0191] ο halogen, CN, OH, -C(O)NR f R g 、-NR f R g wherein R f and R g are each independently H or methyl; and

[0192] ο C 1-2 alkyl, O(C 1-2 alkyl), S(C 1-2 alkyl), SO(C 1-2 alkyl), any of which may optionally be substituted by 1, 2 or 3 substituents selected from halogen, CN and OH;

[0193] ·n is 0;

[0194] ·Z is selected from -NR 10 C(O)-, -C(O)NR 10 -, and -NR 10 C(O)NR 11 -;

[0195] ·L is selected from C 1-3 alkylene and C 2-3 alkenylene;

[0196] ·X is a bond;

[0197] ·p is 0; or p is 1 and R 7 is H;

[0198] ·R 4 is H;

[0199] ·R 5 is selected from: H, -CN, and unsubstituted or substituted with 1, 2 or 3 halogen substituents and / or one -NR 10 R 11 substituent H-substituted C 1 to C 2 alkyl; and

[0200] ·R 6 is H.

[0201] Particularly preferred compounds of the present invention are

[0202] ·5-[[4-[(2-Guanidinoacetyl)amino]-3-(trifluoromethoxy)phenyl]sulfonylamino]thiazole-4-carboxylic acid;

[0203] ·5-[[3-Fluoro-4-[[(2-guanidinoacetyl)amino]methyl]phenyl]sulfonylamino]thiazole-4-carboxylic acid;

[0204] ·5-[[3-Fluoro-4-(guanidinomethyl)phenyl]sulfonylamino]thiazole-4-carboxylic acid;

[0205] ·5-[[3-Fluoro-4-(2-guanidinoethylthiocarbonylamino)phenyl]sulfonylamino]thiazole-4-carboxylic acid;

[0206] ·5-[[4-[2-[(2-Amino-2-imino-ethyl)amino]-2-oxo-ethyl]-3-fluorophenyl]sulfonylamino]thiazole-4-carboxylic acid;

[0207] ·5-[[3-Carbamoyl-4-[(2-guanidinoacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylic acid;

[0208] · 5-[[3-cyano-4-[(2-guanidinoacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylic acid;

[0209] · 5-[[3-fluoro-4-(2-guanidinoethoxycarbonylamino)phenyl]sulfonylamino]thiazole-4-carboxylic acid;

[0210] · 5-[(4-guanidinophenyl)sulfonylamino]thiazole-4-carboxylic acid;

[0211] · 5-[[4-[2-(2-formimidoylhydrazino)-2-oxo-ethyl]-3-fluorophenyl]sulfonylamino]thiazole-4-carboxylic acid;

[0212] · 5-[[3-chloro-4-[(2-guanidinoacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylic acid;

[0213] · 5-[[4-[(2-guanidinoacetyl)amino]-3-methoxyphenyl]sulfonylamino]thiazole-4-carboxylic acid;

[0214] · 5-[[4-[[2-(2-formimidoylhydrazino)acetyl]amino]-3-fluorophenyl]sulfonylamino]thiazole-4-carboxylic acid;

[0215] · 5-[[4-[[(2E)-2-(formimidoylimino)acetyl]amino]-3-fluorophenyl]sulfonylamino]thiazole-4-carboxylic acid;

[0216] · 5-[[4-[[2-(4,5-dihydro-1H-imidazol-2-ylamino)acetyl]amino]-3,5-difluorophenyl]sulfonylamino]thiazole-4-carboxylic acid;

[0217] · 5-[[6-[(2-guanidinoacetyl)amino]pyridazin-3-yl]sulfonylamino]thiazole-4-carboxylic acid;

[0218] · 5-[[4-[(2-amino-2-imino-ethyl)carbamoylamino]-3-fluorophenyl]sulfonylamino]thiazole-4-carboxylic acid;

[0219] · 5-[[3,5-difluoro-4-(guanidinocarbamoylamino)phenyl]sulfonylamino]thiazole-4-carboxylic acid;

[0220] · 5-[[4-[(3-amino-3-imino-propionyl)amino]-3,5-difluorophenyl]sulfonylamino]thiazole-4-carboxylic acid;

[0221] · 5-[[4-[[3-(dimethylamino)-3-imino-propionyl]amino]-3-fluorophenyl]sulfonylamino]thiazole-4-carboxylic acid;

[0222] · 5-[[3-Fluoro-4-[(2-guanidinooxyacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylic acid;

[0223] · 5-[[3-Fluoro-4-[[3-imino-3-(methylamino)propanoyl]amino]phenyl]sulfonylamino]thiazole-4-carboxylic acid;

[0224] · 5-[[4-[3-(4,5-Dihydro-1H-imidazol-2-yl)propanoylamino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylic acid;

[0225] · 5-[[2-[(2-Guanidinoacetyl)amino]thiazol-5-yl]sulfonylamino]thiazole-4-carboxylic acid;

[0226] · 5-[[4-[[2-[(N-cyanoguanidinyl)amino]acetyl]amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylic acid;

[0227] · 5-[[3-Fluoro-4-(guanidinocarbamoylamino)phenyl]sulfonylamino]thiazole-4-carboxylic acid;

[0228] · 5-[[3-Fluoro-4-[[2-(morpholine-4-carboxamidoamino)acetyl]amino]phenyl]sulfonylamino]thiazole-4-carboxylic acid;

[0229] · 5-[[4-[(3-Amino-3-imino-2-methyl-propanoyl)amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylic acid;

[0230] · 5-[[4-[[2-(4,5-Dihydro-1H-imidazol-2-yl)acetyl]amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylic acid;

[0231] · 5-[[4-(Formamidocarbamoylamino)-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylic acid;

[0232] · 5-[[4-[[(2R)-2-Guanidinopropanoyl]amino]phenyl]sulfonylamino]thiazole-4-carboxylic acid;

[0233] · 5-[[3,5-Difluoro-4-[(2-guanidinoacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylic acid;

[0234] · 5-[[4-[(4-Amino-4-imino-butanoyl)amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylic acid;

[0235] · 5-[[4-[[2-(4,5-dihydro-1H-imidazol-2-ylamino)acetyl]amino]-2,5-difluoro-phenyl]sulfonylamino]thiazole-4-carboxylic acid;

[0236] · 5-[[2,5-difluoro-4-[(2-guanidinoacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylic acid;

[0237] · 5-[[3-fluoro-4-[[2-[(N-methylformamidinyl)amino]acetyl]amino]phenyl]sulfonylamino]thiazole-4-carboxylic acid;

[0238] · 5-[[3-fluoro-4-[[2-(2-iminoimidazolidin-1-yl)acetyl]amino]phenyl]sulfonylamino]thiazole-4-carboxylic acid;

[0239] · 5-[[4-[[2-[formamido(methyl)amino]acetyl]amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylic acid;

[0240] · 5-[[4-[[2-[[N-(2-aminoethyl)formamidinyl]amino]acetyl]amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylic acid;

[0241] · 5-[[5-fluoro-6-[(2-guanidinoacetyl)amino]-3-pyridyl]sulfonylamino]thiazole-4-carboxylic acid;

[0242] · 5-[[3-fluoro-4-(3-guanidinopropionylamino)phenyl]sulfonylamino]thiazole-4-carboxylic acid;

[0243] · 5-[[4-[(3-amino-3-imino-propionyl)amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylic acid;

[0244] · 5-[[3,5-difluoro-4-(guanidinocarbamoylamino)phenyl]sulfonylamino]thiazole-4-carboxylic acid;

[0245] · 5-[[3-fluoro-4-[(2-guanidinoacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylic acid; and

[0246] · 5-[[4-[(2-guanidinoacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylic acid;

[0247] and their pharmaceutically acceptable salts.

[0248] Synthesis

[0249] The compounds of the present invention can be prepared by any suitable method. The detailed general synthetic routes of representative compounds of the present invention are set forth below and in the Examples.

[0250] In general, the compounds of the present invention can be prepared in reactions according to the following scheme:

[0251]

[0252] The starting material SM is readily available and can be prepared, for example, using the methods described in WO2014 / 198888. The disclosure of WO2014 / 198849 regarding the formation of compound SM and its analogs is incorporated herein by reference. SM reacts with the sulfonyl chloride derivative of to give the thiazole sulfonamide derivative (A). A reacts with the W-Z-L-X-Pro moiety (B) to give intermediate C. In the above scheme, Q and W are complementary reactive groups that react together to couple A with B to give compound C. For example, Q can be bromine and -Z'-W can be -C(O)NH 2 such that Q and W react together via Buchwald chemistry (especially suitable when n is 0). Alternatively, Q can be -NH 2 and -Z'-W can be -C(O)OH such that Q and W react together in a standard peptide coupling reaction using a reagent such as HATU. Other methods of coupling compounds are well known to those skilled in the art. In compounds B and C, the -NR 7 -Pro moiety represents a protected amine moiety that can be deprotected by standard methods (such as acid-catalyzed deprotection) to give the amine (compound D). Suitable amine protecting groups are well known to those skilled in the art, including the Boc (tert-butoxycarbonyl) protecting group. The amine can then be reacted with a known guanidinylation reagent (such as 1H-pyrazole-1-carboxamide) to form the guanidine group as in compound E. In the compounds of the present invention where p is 0 such that there is an amidino group instead of a guanidine group, the synthesis shown above can be modified such that compound B contains a protected amidino group instead of the protected amine NR 7 -Pro. Suitable amidine protecting groups are well known to those skilled in the art, including the Boc (tert-butoxycarbonyl) protecting group. In these cases, the reaction of A and B gives compound C', which upon deprotection gives the desired amidine product E'. The detailed synthetic routes of exemplary compounds of the present invention are listed below.

[0253] Therapeutic efficacy

[0254] The compounds of the present invention are therapeutically useful. Accordingly, the present invention provides the compounds as described herein for use in medicine. The present invention provides the compounds as described herein for treating a human or animal body. To avoid doubt, the compounds of the present invention can be administered in the form of solvates.

[0255] There is also provided a pharmaceutical composition which comprises a compound of the invention and a pharmaceutically acceptable carrier or diluent, and optionally also comprises an antibiotic agent. Generally, the composition contains up to 85 wt% of the compound of the invention. More typically, the composition contains up to 50 wt% of the compound of the invention. Preferred pharmaceutical compositions are sterile and pyrogen-free. Further, when the pharmaceutical composition provided by the invention contains an optically active compound of the invention, the compound of the invention is generally a substantially pure optical isomer.

[0256] The composition of the invention may be provided as a kit which comprises instructions enabling the kit to be used in the methods described herein or details regarding which subjects the methods are applicable to.

[0257] As described above, the compounds of the invention can be used for treating or preventing bacterial infections. In particular, they are inhibitors of metallo-β-lactamase (MBL), and thus can be used to remove or reduce the resistance of Gram-negative bacteria to antibiotics.

[0258] The compounds of the invention can be used as an independent therapeutic agent. For example, the compounds of the invention can be used as an independent adjuvant in antibacterial therapy (such as in a chemotherapy regimen). Alternatively, the compounds of the invention can be used in combination with an antibiotic agent to enhance the action of the antibiotic agent. The compounds of the invention can be particularly used for treating or preventing bacterial infections caused by bacteria resistant to treatment with the antibiotic agent alone, especially in cases where the resistance is caused by the presence of metallo-β-lactamase and / or serine-β-lactamase. Treatment or prevention of such infections with β-lactam antibiotics alone may not be successful.

[0259] Accordingly, the invention also provides a product which comprises (i) a compound of the invention as described herein and (ii) an antibiotic agent. The compound of the invention and the antibiotic agent can be provided in a single formulation, or the compound of the invention and the antibiotic agent can be formulated separately. In the case of separate formulation, the two agents can be administered simultaneously or separately. They can be provided in the form of a kit, optionally together with instructions for their administration. The product may also be referred to herein as a combination or a pharmaceutical combination.

[0260] When formulated together, the two active agents can be provided as a pharmaceutical composition which comprises (i) a compound of the invention as described herein and (ii) an additional antibacterial compound; and (iii) a pharmaceutically acceptable carrier or diluent.

[0261] Preferably, the antibiotic agent is a β-lactam antibiotic. More preferably, the antibiotic agent is a β-lactam antibiotic selected from the group consisting of carbapenems, penicillins, cephalosporins, and penems. Examples of carbapenem antibiotics 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 ceftobiprole. Examples of penems include faropenem. Other antibiotic agents 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, and most preferably meropenem.

[0262] The product of the present invention may further comprise a serine-β-lactamase (SBL) inhibitor. Accordingly, the present invention also provides a product comprising (i) a compound of the present invention; (ii) a serine-β-lactamase (SBL) inhibitor; and (iii) an antibiotic agent. These products are referred to herein as "triple combinations". The triple combinations comprise the three active agents (i) to (iii) described above, but may also comprise other active agents if desired.

[0263] In the triple combinations of the present invention, the compound of the present invention, the SBL inhibitor, and the antibiotic agent may each be provided in a single formulation, or they may be formulated separately. Alternatively, two of the components may be provided in a single formulation, while the remaining component is provided separately. In other words, the compound of the present invention may be formulated together with the SBL inhibitor and the antibiotic agent; or, the compound of the present invention may be formulated together with the SBL inhibitor, while the antibiotic agent is provided separately; or, the compound of the present invention may be formulated together with the antibiotic agent, while the SBL inhibitor is provided separately; or, the SBL inhibitor may be formulated together with the antibiotic agent, while the compound of the present invention is provided separately; or, the compound of the present invention, the SBL inhibitor, and the antibiotic agent may each be formulated separately. In the case of separate formulation, the components of the triple combination may be administered simultaneously or separately. They may be provided in the form of a kit, optionally together with instructions for their administration.

[0264] When two or more active agents are formulated together, the two or more active agents may be provided as a pharmaceutical composition comprising (i) a compound of the present invention as described herein; (ii) a pharmaceutically acceptable carrier or diluent; and (iii) one or both of an antibiotic agent and (iv) a serine-β-lactamase (SBL) inhibitor.

[0265] In the triple combination of the present invention, the SBL inhibitor is a compound of formula (II) or a pharmaceutically acceptable salt thereof,

[0266]

[0267] wherein,

[0268] οG is selected from -CN and -C(O)NR j R k ;

[0269] οR k is selected from -W and -QW; where W is selected from 5- to 6-membered heterocyclic groups, R j and -N(R j ) 2 ; Q is selected from -NR j C(O)-, -C(O)-NR j -, C 1-3 alkylene, -OC 1-3 alkylene and -N(R j )-C 1-3 alkylene;

[0270] οEach R j is selected from H and unsubstituted C 1-3 alkyl, preferably H.

[0271] In formula (II), when W is a 5- to 6-membered heterocyclic group, W is preferably a 6-membered heterocyclic ring containing a nitrogen atom; more preferably W is piperidinyl. Preferably, in formula (II), W is selected from 5- to 6-membered heterocyclic groups and -N(R j ) 2 , more preferably W is selected from piperidinyl and NH 2 . In formula (II), Q is preferably selected from -NR j C(O)- and -OC 1-3 alkylene. Preferably, in formula (II), each R j is H. Thus, the preferred definition of G in formula (II) is -CN and -C(O)NHR k , where R k is selected from -W and -QW; where W is selected from 5- to 6-membered heterocyclic groups and -NH 2 , the 5- to 6-membered heterocyclic group is preferably pyridinyl; Q is selected from -NHC(O)- and -OC 1-3 alkylene.

[0272] More preferably, in the pharmaceutical combination of the present invention, the SBL inhibitor is selected from WCK4234, avibactam, relebactam, zidebactam, and nacubactam, or a pharmaceutically acceptable salt thereof. The structures of WCK4234, avibactam, relebactam, zidebactam, and nacubactam 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. For example, WCK4234 and its synthesis are described in WO2013 / 038330 and WO2015 / 114595. Ball, M. et al, Org. Process Res. Dev., 2016, 20(10), pp 1799–1805 and US2012 / 323010 describe avibactam and its synthesis. WO2009 / 091856 describes relebactam and its synthesis. WO 2015 / 110885 describes zidebactam and its synthesis. WO2014 / 091268 and US2016 / 272641 describe Nacubactam and its synthesis.

[0273]

[0274]

[0275] More preferably, in the pharmaceutical combination of the present invention, the SBL inhibitor is WCK4234 or a pharmaceutically acceptable salt thereof. More preferably, the SBL inhibitor is WCK4234 or its sodium salt. The process for preparing the sodium salt of WCK4234 is described in WO 2015 / 114595.

[0276] In the triple combination of the present invention, the antibiotic agent can be any antibiotic agent disclosed herein. Preferably, in the pharmaceutical combination of the present invention, the antibiotic agent is a β-lactam antibiotic. Preferably, the β-lactam antibiotic is selected from carbapenems, penicillins, cephalosporins, and penems, more preferably the β-lactam antibiotic is a carbapenem antibiotic, preferably imipenem or meropenem, and most preferably meropenem.

[0277] Therefore, most preferably, the pharmaceutical combination of the present invention comprises (i) a compound of the present invention; (ii) an SBL inhibitor selected from WCK4234, avibactam, relebactam, zidebactam, and nacubactam and pharmaceutically acceptable salts thereof, preferably WCK4234 or a pharmaceutically acceptable salt thereof; (iii) a carbapenem antibiotic, preferably meropenem.

[0278] The compounds of the present invention can also be used for treating or preventing bacterial infections. Accordingly, the present invention provides the compounds of the present invention for use in medicine. The present invention also provides the use of the compounds of the present invention in the preparation of medicaments. The present invention also provides compositions and products comprising the compounds of the present invention as described herein. Such compositions and products can also be used for treating or preventing bacterial infections. Accordingly, the present invention provides compositions or products as defined herein for use in medicine. The present invention also provides the use of the compositions or products of the present invention in the preparation of medicaments.

[0279] As explained above, the compounds, compositions and products of the present invention can be used for treating or preventing bacterial infections. Accordingly, the present invention also provides a method for treating or preventing bacterial infections in a subject, which method comprises administering to the subject an effective amount of a compound, composition or product as described herein. Also provided are the compounds, compositions or products of the present invention as described herein for use in the preparation of a medicament for treating or preventing bacterial infections; the compounds of the present invention are generally used in combination with antibiotic agents.

[0280] As further explained above, the compounds of the present invention can be used in combination with additional antibacterial compounds. Accordingly, the present invention provides the compounds of the present invention for use in treating or preventing bacterial infections, wherein such use comprises co-administering the compounds of the present invention with an additional antibacterial compound. The present invention also provides the use of the compounds of the present invention in the preparation of a medicament for treating or preventing bacterial infections by co-administering the compounds of the present invention with an additional antibacterial compound. The present invention also provides a method for treating or preventing bacterial infections by co-administering the compounds of the present invention and an additional antibacterial compound to a subject in need thereof. The additional antibacterial compound is preferably an antibacterial compound as described herein; more preferably a β-lactam antibiotic as described herein.

[0281] The compounds of the present invention can also be used in combination with serine-β-lactamase (SBL) inhibitors and antibiotic agents, i.e., as a "triple combination". Accordingly, the present invention provides the compounds of the present invention for the treatment or prevention of bacterial infections, wherein such use comprises co-administering (i) a compound of the present invention and (ii) a serine-β-lactamase (SBL) inhibitor and (iii) an antibiotic agent. There is also provided an antibiotic agent for the treatment or prevention of bacterial infections by co-administering with a compound of the present invention and an optional SBL inhibitor. There is also provided an SBL inhibitor for the treatment or prevention of bacterial infections by co-administering with a compound of the present invention and an optional antibiotic agent. The present invention also provides the use of a compound of the present invention in the preparation of a medicament for the treatment or prevention of bacterial infections by co-administering (i) a compound of the present invention with (ii) a serine-β-lactamase (SBL) inhibitor and (iii) an antibiotic agent. There is also provided the use of an antibiotic agent in the preparation of a medicament for the treatment or prevention of bacterial infections by co-administering with a compound of the present invention and an optional SBL inhibitor. There is also provided the use of an SBL inhibitor in the preparation of a medicament for the treatment or prevention of bacterial infections by co-administering with a compound of the present invention and an optional antibiotic agent. The present invention also provides a method for treating or preventing bacterial infections by co-administering (i) a compound of the present invention, (ii) a serine-β-lactamase (SBL) inhibitor, and / or (iii) an antibiotic agent to a subject in need thereof. The serine-β-lactamase (SBL) inhibitor is preferably a serine-β-lactamase (SBL) inhibitor as described herein. The antibiotic agent is preferably an antibacterial compound as described herein; more preferably a β-lactam antibiotic as described herein.

[0282] In one aspect, the subject is a mammal, particularly a human. However, the subject may be non-human. Preferred non-human animals include, but are not limited to, primates such as marmosets or monkeys, commercially farmed animals (such as horses, cows, sheep or pigs), and pets (such as dogs, cats, mice, rats, guinea pigs, ferrets, gerbils or hamsters). The subject can be any animal capable of being infected by bacteria.

[0283] The compounds, compositions and combinations described herein can be used to treat bacterial infections that occur after recurrence following antibiotic treatment. Accordingly, the compounds, compositions and combinations can be used to treat patients who have previously received antibiotic treatment for a (similarly experienced) bacterial infection.

[0284] The bacteria causing the infection can be any bacteria that express a metallo-β-lactamase or an analogue thereof. Generally, the bacteria causing the infection express an MBL enzyme. The bacteria are generally Gram-negative. The bacteria can in particular be pathogenic bacteria. Generally, the bacterial infections to be treated with the compounds of the present invention are resistant to conventional antibiotic treatment when using conventional antibiotics alone.

[0285] The compound of general formula (I) can be used to remove antibiotic-resistant Gram-negative bacteria, which are bacteria that produce metallo-β-lactamases. The bacteria that produce metallo-β-lactamases can be metallo-β-lactamases of subclasses B1, B2 or B3, such as IMP-type (including IMP-1), VIM-type (including VIM-1 and VIM-2) and NDM-type (including NDM-1) enzymes. 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; most typically, 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.

[0286] The bacterial infection can be caused by bacteria from the Enterobacteriaceae, Pseudomonadaceae and / or Moraxellaceae. More commonly, the bacterial infection is caused by bacteria from the Enterobacteriaceae and / or Pseudomonadaceae. Most typically, the bacterial infection is caused by bacteria from the Enterobacteriaceae. The bacterial infection can be caused by Pseudomonas (such as Pseudomonas aeruginosa, Pseudomonas oryzihabitans or Pseudomonas plecoglossicida), Klebsiella, Escherichia, Acinetobacter or Burkholderia. For example, the 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 bacteria can be opportunistic pathogens.

[0287] The compounds, compositions and products of the present invention can be used to prevent or treat infections of the following strains:

[0288] NTBC020 (Escherichia coli strain expressing NDM-1, TEM-1, and CTX-M-15), NTBC035-2 (Klebsiella pneumoniae strain expressing NDM-1, CMY-4, and SHV-11), NTBC104-1 (Klebsiella pneumoniae strain expressing NDM-1 and SHV-11), NTBC123 (Klebsiella pneumoniae strain expressing NDM-1), NTBC062 (Klebsiella pneumoniae strain expressing IMP-1 and TEM-1), NTBC024 (Klebsiella pneumoniae strain expressing VIM-19, TEM-1, and CTX-M-3), NTBC042 (Escherichia coli strain expressing VIM-1, TEM-1, CTX-M-15, and SHV-12), NTBC055 (Escherichia coli strain expressing VIM-1), and NTBC039 (Klebsiella oxytoca strain expressing IMP-28).

[0289] The compounds, compositions, and products of the present invention can also be used for preventing or treating infections by the following strains. The triple combination is particularly useful for preventing or treating infections by these strains:

[0290] NTBC019 (Klebsiella pneumoniae strain expressing NDM-1, CTX-M-15, and OXA-181), NTBC185 (Klebsiella pneumoniae expressing SHV-OSBL, TEM-OSBL, NDM-1, and OXA-48), NTBC186 (Klebsiella pneumoniae strain expressing ACT-TYPE, VIM-1, and OXA-48), NTBC187 (Klebsiella pneumoniae strain expressing SHV-OSBL, NDM-1, and OXA-48), and NTBC188 (Klebsiella pneumoniae strain expressing NDM-1 and KPC-2).

[0291] The compounds, compositions, or combinations of the present invention can be used for treating or preventing infections and diseases caused by any one of the above bacteria or combinations of the above bacteria. In particular, the compounds, compositions, or combinations of the present invention can be used for treating or preventing pneumonia. The compound, composition, or combination can also be used for treating septic shock, urinary tract infections, and gastrointestinal, skin, or soft tissue infections.

[0292] The compounds, compositions, or combinations of the present invention can be used for treating patients infected with carbapenem-resistant Enterobacteriaceae (CRE). CRE can be found in the community or in hospitals and other institutions, which are usually associated with long-term patients and those undergoing significant medical interventions such as those typically carried out in intensive care units (ICUs).

[0293] The compounds, compositions or combinations of the present invention can be administered to a subject to prevent the onset or recurrence of one or more symptoms of a bacterial infection. This is the prophylactic method. In this embodiment, the subject can be asymptomatic. The subject is typically a subject who has been exposed to bacteria. A prophylactically effective amount of the agent or formulation is administered to the subject. A prophylactically effective amount is an amount that prevents the onset of one or more symptoms of a bacterial infection.

[0294] The compounds, compositions or combinations of the present invention can be administered to a subject to treat one or more symptoms of a bacterial infection. In this embodiment, the subject is typically symptomatic. A therapeutically effective amount of the agent or formulation is administered to such a subject. A therapeutically effective amount is an amount that effectively ameliorates one or more symptoms of the condition.

[0295] The compounds, compositions or combinations of the present invention can be administered in a variety of dosage forms. Thus, the compounds, compositions or combinations of the present invention can be administered orally, for example, as tablets, lozenges, troches, aqueous or oily suspensions, dispersible powders or granules. The pharmaceutical compositions of the present invention can also be administered parenterally, whether subcutaneously, intravenously, intramuscularly, intrasternal, transdermally or by infusion techniques. The compound, composition or combination can also be administered as a suppository. Preferably, the compound, composition or combination can be administered by inhalation (nebulization) or intravenously, most preferably by inhalation (nebulization) administration.

[0296] The compounds, compositions or combinations of the present invention are generally formulated for administration with a pharmaceutically acceptable carrier or diluent. For example, solid oral dosage forms can contain the active compound and a diluent (such as lactose, dextrose, sucrose, cellulose, corn starch or potato starch), a lubricant (such as silica, talc, stearic acid, magnesium stearate or calcium stearate, and / or polyethylene glycol), a binder (such as starch, gum arabic, gelatin, methylcellulose, carboxymethylcellulose or polyvinylpyrrolidone), a disintegrant (such as starch, alginic acid, alginates or sodium starch glycolate), an effervescent mixture, a dye, a sweetening agent, a wetting agent (such as lecithin, polysorbates, dodecyl sulfates), and non-toxic and pharmacologically inert substances commonly used in pharmaceutical formulations. These pharmaceutical formulations can be prepared in known ways, for example, by mixing, granulating, tableting, sugar coating or film coating processes.

[0297] The compounds, compositions or combinations of the present invention can be formulated into solutions or suspensions for administration by inhalation (nebulization). The compounds, compositions or combinations of the present invention can be administered by a metered-dose inhaler (MDI) or a nebulizer (e.g., an electronic or jet nebulizer). Alternatively, the compounds, compositions or combinations of the present invention can be formulated into powdered drugs for administration by inhalation, and such formulations can be administered from a dry powder inhaler (DPI). The compounds, compositions or combinations of the present invention can be delivered in particulate form when formulated for administration by inhalation, and the mass median aerodynamic diameter (MMAD) of the particles is from 1 μm to 100 μm, preferably from 1 μm to 50 μm, more preferably from 1 μm to 20 μm, such as from 3 μm to 10 μm, such as from 4 μm to 6 μm. When the compounds, compositions or combinations of the present invention are delivered as an aerosolized aerosol, the reference to particle size defines the MMAD of the aerosol droplets. The MMAD can be measured by any suitable technique such as laser diffraction.

[0298] Liquid dispersions for oral administration can be syrups, emulsions and suspensions. Syrups can contain a carrier such as sucrose or sucrose with glycerol and / or mannitol and / or sorbitol.

[0299] Suspensions and emulsions can contain, for example, natural gums, agar, sodium alginate, pectin, methylcellulose, carboxymethylcellulose or polyvinyl alcohol as carriers. Suspensions or solutions for intramuscular injection or inhalation can contain the active compound and a pharmaceutically acceptable carrier such as sterile water, olive oil, ethyl oleate, a diol (e.g., propylene glycol), and, if desired, an appropriate amount of lidocaine hydrochloride.

[0300] Solutions for inhalation, injection or infusion can contain, for example, sterile water as a carrier, or preferably they can be in the form of a sterile aqueous isotonic saline solution. Pharmaceutical compositions suitable for delivery by needleless injection (e.g., transdermal) can also be used.

[0301] A therapeutically or prophylactically effective amount of the compound of the present invention is administered to a subject. The dosage can be determined according to various parameters, especially according to the compound used, the age, weight and condition of the subject to be treated, the route of administration, and the desired regimen. Similarly, the doctor will be able to determine the route of administration and dosage required for any particular subject. Depending on the activity of the specific inhibitor, the age, weight and condition of the subject to be treated, the type and severity of the disease, and the frequency and route of administration, a typical daily dose is about 0.01 mg to 100 mg / kg of body weight, preferably about 0.1 mg to 50 mg per kilogram of body weight, such as about 1 mg to 10 mg per kilogram of body weight. Preferably, the daily dose level is from 5 mg to 2 g.

[0302] When the compounds of the present invention are administered to a subject in combination with another active agent (e.g., in the form of a pharmaceutical combination comprising an antibiotic agent and an optional SBL inhibitor), the dose of the other active agent (e.g., SBL inhibitor and / or antibiotic agent) can be determined as described above. The dose can be determined according to various parameters, especially according to the agent used, the age, weight and condition of the subject to be treated, the route of administration, and the desired regimen. Again, the doctor will be able to determine the route of administration and dose required for any particular subject. Depending on the activity of the specific inhibitor, the age, weight and condition of the subject to be treated, the type and severity of the disease, and the frequency and route of administration, the typical daily dose is about 0.01 mg to 100 mg per kg of body weight, preferably about 0.1 mg to 50 mg per kg of body weight, e.g., about 1 to 10 mg per kg of body weight. Preferably, the daily dose level is 5 mg to 2 g.

[0303] The antibacterial properties of the compounds described herein mean that they can also be used for the in vitro treatment of bacterial infections, i.e., not for the treatment of human or animal subjects. Accordingly, the present invention also provides a cleaning composition comprising a thiazole derivative of formula (I) or a salt thereof. The cleaning composition may further comprise, for example, a detergent, a surfactant (including ionic and non-ionic surfactants), a diluent, a bleach (including hypochlorites (such as sodium hypochlorite or calcium hypochlorite), chlorine, chlorine dioxide, hydrogen peroxide or its adducts, sodium perborate, and sodium percarbonate), an alcohol (e.g., ethanol or isopropanol), or a disinfectant. Generally, the disinfectant may be selected from benzyl-4-chlorophenol, amylphenol, phenylphenol, glutaraldehyde, alkyldimethylbenzylammonium chloride, alkyldimethylethylbenzylammonium chloride, iodine, peracetic acid, and chlorine dioxide. Generally, the detergent may be an alkaline detergent (e.g., sodium hydroxide, sodium metasilicate, or sodium carbonate) or an acidic detergent (e.g., hydrochloric acid, nitric acid, sulfuric acid, phosphoric acid, citric acid, or tartaric acid).

[0304] The following examples illustrate the invention. However, they do not limit the invention in any way. In this regard, it is important to understand that the specific tests used in the examples section are only designed to provide an indication of biological activity. There are many tests available for determining biological activity, and thus a negative result in any one specific test is not conclusive.

[0305] Experimental details

[0306] General synthetic methods

[0307] There are several related synthetic methods for this type of compound described by formula 1, and as described below, where R refers to any substituent on the benzene ring.

[0308]

[0309] The preparation of the key thiazole intermediate tert-butyl 5-{[(4-methoxyphenyl)methyl]amino}-1,3-thiazole-4-carboxylate has been previously described (WO2014 / 198849) and is readily prepared on a 100 g scale. Its reaction with various arylsulfonyl chlorides gives sulfonamide intermediates such as [A] by using a base catalyst (e.g., pyridine, triethylamine or sodium hydride). Other forms of thiazole starting materials (e.g., R1 = ethyl; R2 = H) are also readily available or even commercially available. Many of the compounds described herein can be obtained from the bromophenylsulfonamide [A] via a standard Buchwald reaction with a protected glycinamide such as [B]. Acid-catalyzed global deprotection shows that if desired, the primary amine [C] can be converted to the guanidine [D] using a guanidinylation reagent such as 1H-pyrazole-1-carboxamide (Scheme 1).

[0310]

[0311] Alternatively, instead of forming the aryl-nitrogen bond by using Buchwald chemistry on the aryl bromide, certain aniline intermediates such as [E] can be reacted with an N-protected glycine using a standard peptide coupling reagent such as HATU (Scheme 2). Deprotection and guanidinylation again give [C] and [D], respectively. The aniline [E] can be obtained from the corresponding nitro compound by standard reduction or from a bromo intermediate using ammonia via a Buchwald reaction (e.g., see Scheme 4).

[0312]

[0313] In some cases, such as when the substituents on the aryl ring are particularly electron-withdrawing, both Buchwald amidation and formation of the amide using a protected glycine derivative are unsuccessful. For these cases, the aniline must be reacted with highly reactive chloroacetyl chloride to give the intermediate [F]. This is then displaced with sodium azide to give the azidoacetamide [G], which can be reduced with a standard reducing agent to give [C] and [D] in a conventional manner (Scheme 3).

[0314]

[0315] Certain urea derivatives require custom synthesis (Scheme 4). For example, a Buchwald reaction of a typical bromoarylsulfonamide with ammonia, which itself serves as the nitrogen-containing component, gives the corresponding aniline. This aniline is activated with 4-nitrophenyl chloroformate to give [H], and the coupling product [J] can be obtained by reacting [H] with a BOC-protected hydrazine [I] via the intermediacy of the isocyanate derived from [H]. Mild acid treatment removes the BOC group that can be guanidinylated to give the protected guanidine functionality. Then, global deprotection of the BOC, p-methoxybenzyl and tert-butyl ester groups gives the guanidine [K].

[0316]

[0317] Some analogs require a key glyoxylamide intermediate [M], which is synthesized by reacting a conventional aniline with 0.5 equivalent of fumaryl chloride to obtain a symmetric diamide [L]. Ozonolysis is carried out to obtain the unstable glyoxylamide [M], which can react with various nucleophiles including bis-BOC protected aminoguanidine to obtain [N]. Then, full deprotection is carried out in a conventional manner to obtain the corresponding imine [O] (Scheme 5).

[0318] Abbreviations

[0319] ACN Acetonitrile

[0320] AcOH Acetic acid

[0321] Ag(OTf) Silver trifluoromethanesulfonate

[0322] AIBN Azobisisobutyronitrile

[0323] Boc tert-Butoxycarbonyl

[0324] Boc2O Di-tert-butyl dicarbonate

[0325] Cs 2 CO 3 Cesium carbonate

[0326] CFU Colony forming unit

[0327] CuI Copper iodide

[0328] DCM Dichloromethane

[0329] DIPEA N,N-Diisopropylethylamine

[0330] DMAP 4-Dimethylaminopyridine

[0331] DMF N,N-Dimethylformamide

[0332] DMS Dimethyl sulfide

[0333] DMSO Dimethyl sulfoxide

[0334] dppf 1,1'-Bis(diphenylphosphino)ferrocene

[0335] EDC.HCl N-(3-Dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride

[0336] EtOAc Ethyl acetate

[0337] EtOH Ethanol

[0338] Et3N Triethylamine

[0339] HATU 1-[Bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate

[0340] HCl Hydrochloric acid

[0341] HOBt Hydroxybenzotriazole

[0342] H2SO4 Sulfuric acid

[0343] IPA Isopropyl alcohol

[0344] Km Michaelis constant

[0345] MeI Methyl iodide

[0346] MeOH Methanol

[0347] NBS N-Bromosuccinimide

[0348] Na2CO3 Sodium carbonate

[0349] Na2SO4 Sodium sulfate

[0350] Pd 2 (dba) 3 Tris(dibenzylideneacetone)dipalladium(0)

[0351] PdCl 2 (PPh3)2 Bis(triphenylphosphine)palladium(II) dichloride

[0352] PdCl 2 (dppf) [1,1'-Bis(diphenylphosphino)ferrocene]palladium(II) dichloride

[0353] PMB p-Methoxybenzyl

[0354] TEA Triethylamine

[0355] TES Triethylsilyl

[0356] TMSOK Potassium trimethylsilanolate

[0357] TFA Trifluoroacetic acid

[0358] TMSOTf Trimethylsilyl trifluoromethanesulfonate

[0359] TFA Trifluoroacetic acid

[0360] THF Tetrahydrofuran

[0361] T3P Propylphosphinic anhydride

[0362] RT Room temperature

[0363] The structure of palladium(II) chloride (1:1 MTBE adduct) of (RuPhos)phenethylamine (RuPhos Pd G1 complex) used in the Buchwald coupling step is shown below.

[0364]

[0365] Example

[0366] General techniques

[0367] 1H NMR spectra (δ in ppm) were reported in DMSO-d6 solution at 300 or 400 MHz using chloroform as the reference standard (7.25 ppm). When reporting peak multiplicities, the following abbreviations were used: s (singlet), d (doublet), t (triplet), m (multiplet), bs (broad singlet), dd (doublet of doublets), dt (doublet of triplets), q (quartet). Coupling constants were reported in hertz (Hz) when given.

[0368] The term “purified by prep hplc (MDAP)” refers to the purification of a compound using a mass-directed automated purification system, eluting on an Agilent 1260 infinity machine with an XSelect CHS Prep C18 column, using a 0.1% formic acid in water / acetonitrile solution and detected by Quadruploe LC / MS.

[0369] Example 1

[0370] tert-Butyl 5-[(4-methoxyphenyl)methylamino]thiazole-4-carboxylate

[0371] (Key intermediate - 1)

[0372]

[0373] A suspension of potassium tert-butoxide (874 mg, 7.79 mmol) in anhydrous tetrahydrofuran (10 mL) was stirred vigorously 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. At room temperature, a solution of 4-methoxybenzyl isothiocyanate (1.27 g, 7.08 mmol) in anhydrous tetrahydrofuran (5 mL) was added dropwise thereto. After 2 hours, the solution was poured into saturated NaHCO3 solution and extracted with ethyl acetate. The organic layer was washed with Na 2 SO4 Dry, filter and concentrate in vacuo to dryness. Purify the residue by silica gel chromatography (eluting with 0 to 50% ethyl acetate / cyclohexane) to give the title product (852 mg) as a pale yellow solid.

[0374] 1H NMR(CDCl 3 ) 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).

[0375] M / z 321 (M + H) +

[0376] Example 2

[0377] 5 - [[3,5 - Difluoro - 4 - [(2 - guanidinoacetyl)amino]phenyl]sulfonylamino]thiazole - 4 - carboxylic acid

[0378]

[0379] a. tert - Butyl 5 - [(4 - bromo - 3,5 - difluorophenyl)sulfonyl - [(4 - methoxyphenyl)methyl]amino]thiazole - 4 - carboxylate

[0380]

[0381] Under 0 °C and an argon atmosphere, add a solution of tert - butyl 5 - [(4 - methoxyphenyl)methylamino]thiazole - 4 - carboxylate (1 g, 3.12 mmol, 1 equiv) in THF (15 mL) to a suspension of NaH in THF (10 mL). After 30 minutes, under 0 °C and an argon atmosphere, add a solution of 4 - bromo - 3,5 - difluorobenzenesulfonyl chloride (1.0 g, 3.43 mmol, 1.1 equiv) in THF (15 mL). Stir the resulting reaction mixture at room temperature for 1 h, quench with ice - water (20 mL) and extract with ethyl acetate (2×20 mL). Dry the combined organic layers over Na 2 SO 4 Dry, filter and concentrate. Purify the crude material by trituration with diethyl ether (2×5 mL) to give a pale yellow solid (800 mg, 44%).

[0382] M / z 577.0 (M + H) +

[0383] b. tert - Butyl 5 - [[4 - [[2 - (tert - butoxycarbonylamino)acetyl]amino]-3,5 - difluorophenyl]sulfonyl - [(4 - methoxyphenyl)methyl]amino]thiazole - 4 - carboxylate

[0384]

[0385] Purge a solution of tert-butyl 5-[(4-bromo-3,5-difluorophenyl)sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate (100 mg, 0.173 mmol, 1 equiv) in 1,4-dioxane (5 mL) with argon for 15 min. Then add tert-butyl N-(2-amino-2-oxoethyl)carbamate (45 mg, 0.26 mmol, 1.5 equiv), K 3 PO 4 (110 mg, 0.521 mmol, 3 equiv), Pd 2 (dba) 3 (16 mg, 0.17 mmol, 0.1 equiv) and Xantphos (30 mg, 0.052 mmol, 0.3 equiv) under an argon atmosphere. Heat the resulting reaction mixture in a sealed vial to 85 °C and hold for 16 h. Cool the temperature to room temperature, filter the reaction mixture through a pad of diatomaceous earth, and wash the pad with EtOAc (2 × 5 mL). Concentrate the organic layer under reduced pressure. Dissolve the resulting crude compound in ethyl acetate (25 mL), wash with water (10 mL) and brine solution (10 mL). Dry the organic layer over Na 2 SO 4 , filter and concentrate. Purify the crude material by flash chromatography (eluting with a petroleum ether solution of 55% ethyl acetate) to give a pale yellow solid (60 mg, 51%).

[0386] M / z 669.5 (M+H) +

[0387] c. 5-[[4-[(2-Aminoacetyl)amino]-3,5-difluorophenyl]sulfonylamino]thiazole-4-carboxylic acid trifluoroacetate

[0388]

[0389] Add TFA (4 mL) to tert-butyl 5-[[4-[[2-(tert-butoxycarbonylamino)acetyl]amino]-3,5-difluorophenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate (300 mg, 0.448 mmol, 1 equiv) at room temperature and stir for 4 h. Evaporate TFA under reduced pressure. Triturate the resulting crude product with diethyl ether (2 × 5 mL) and dry under high vacuum to give a pale yellow solid (150 mg, 85%).

[0390] M / z 393.3 (M+H) +

[0391] d. 5-[[3,5-difluoro-4-[(2-guanidinoacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylic acid

[0392]

[0393] At room temperature, pyrazole-1-carboxamidine hydrochloride (84 mg, 0.573 mmol, 1.5 equiv) and DIPEA (0.3 mL, 1.91 mmol, 5 equiv) were added to a stirred solution of 5-[[4-[(2-aminoacetyl)amino]-3,5-difluoro-phenyl]sulfonylamino]thiazole-4-carboxylic acid trifluoroacetate (150 mg, 0.382 mmol, 1 equiv) in DMF (5 mL). The resulting reaction mixture was stirred at room temperature for 16 h and concentrated under reduced pressure. Water (5 mL) was added to the residue, the precipitate was filtered and washed with diethyl ether (2 × 5 mL). The crude product was purified by preparative HPLC to give the title compound as a white solid (47 mg, 28%).

[0394] 1 H NMR (400 MHz, DMSO-d 6 ) 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).

[0395] M / z 434.9 (M+H) +

[0396] LC-MS conditions:

[0397] Column: Acquity BEH C18 (50 mm x 2.1 mm, 1.7 um);

[0398] Mobile phase: A: aqueous solution of 0.05% formic acid;

[0399] B: ACN solution of 0.05% formic acid

[0400] Time (min) / % B: 0 / 3, 0.4 / 3, 2 / 98, 3.4 / 98, 3.5 / 3, 4 / 3;

[0401] Column temperature: 35 °C, flow rate: 0.6 mL / min

[0402] Prep.HPLC conditions:

[0403] Column: Symmetry C18 (300 × 19) mm, 7u;

[0404] Mobile phase: (A) 0.1% formic acid (B) acetonitrile;

[0405] Flow rate: 19 mL / min;

[0406] Gradient (T / %B): 0 / 5, 1 / 5, 8 / 20, 11 / 20, 11.02 / 99, 12 / 99, 12.1 / 5, 15 / 5;

[0407] Solubility: ACN + H 2 O + THF + FA

[0408] Example 3

[0409] 5-[[5-Fluoro-6-[(2-guanidinoacetyl)amino]-3-pyridyl]sulfonylamino]thiazole-4-carboxylic acid

[0410]

[0411] a. 5-Fluoro-6-hydroxy-pyridine-3-sulfonyl chloride

[0412]

[0413] At 0 °C, 3-fluoropyridin-2-ol (2 g, 17.6 mmol) was added to chlorosulfonic acid (20 mL, 300.3 mmol). The reaction mixture was stirred at 160 °C for 2 h, cooled to room temperature and slowly poured into ice water (50 mL). The aqueous layer was extracted with EtOAc (3 × 50 mL). The organic layer was dried over Na 2 SO 4 dried, filtered and concentrated under reduced pressure. The crude compound was triturated with n-pentane (2 × 50 mL) to give an off-white solid (2.7 g, 72%).

[0414] M / z 212.11 (M + H) +

[0415] b. 6-Chloro-5-fluoro-pyridine-3-sulfonyl chloride

[0416]

[0417] At 0 °C, thionyl chloride (5 mL, 68.9 mmol) was added to a solution of 5-fluoro-6-hydroxy-pyridine-3-sulfonyl chloride (1 g, 4.73 mmol) in toluene (25 mL). Then DMF (0.2 mL) was slowly added at 0 °C. The reaction mixture was refluxed for 3 h, cooled to room temperature and concentrated under reduced pressure. The resulting crude material was co-distilled with toluene (2 × 25 mL) to give a pale yellow liquid which was used in the next step without further purification (0.9 g, crude).

[0418] 1 H NMR (400 MHz, DMSO-d 6 ) δ 8.83 (1H, m), 8.04 (1H, m).

[0419] c. tert-Butyl 5-[(6-chloro-5-fluoro-3-pyridinyl)sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

[0420]

[0421] At 0 °C under an argon atmosphere, 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 suspension of NaH (1.12 g, 46.8 mmol) in THF (10 mL). After 15 minutes, at 0 °C under an argon atmosphere, a solution of 6-chloro-5-fluoro-pyridine-3-sulfonyl chloride (1.6 g, 7.0 mmol) in THF (15 mL) was added to the above reaction mixture. The resulting reaction mixture was stirred at room temperature for 0.5 h, quenched with ice water (20 mL), and extracted with ethyl acetate (3 × 20 mL). The combined organic layers were dried over Na 2 SO 4 dried, filtered, and concentrated. The crude compound was purified by flash chromatography (eluting with a petroleum ether solution of 10% to 15% ethyl acetate) to give a yellow oil (1.3 g, 54%).

[0422] M / z 514.27 (M + H) +

[0423] d. tert-Butyl 5-[[6-[[2-(tert-butoxycarbonylamino)acetyl]amino]-5-fluoro-3-pyridinyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

[0424]

[0425] A solution of tert-butyl 5-[(6-chloro-5-fluoro-3-pyridinyl)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 minutes. Then, under an argon atmosphere, tert-butyl N-(2-amino-2-oxo-ethyl)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). The resulting reaction mixture was heated to 70 °C in a sealed tube for 0.5 h, cooled to room temperature, filtered through a pad of diatomaceous earth, and the pad was washed with ethyl acetate (2 × 3 mL). The organic layer was concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with a petroleum ether solution of 50% ethyl acetate) to give a pale yellow solid (75 mg, 39%).

[0426] M / z 652.41 (M+H) +

[0427] e. 5-[[6-[(2-Aminoacetyl)amino]-5-fluoro-3-pyridinyl]sulfonylamino]thiazole-4-carboxylic acid trifluoroacetate

[0428]

[0429] At 0 °C, TFA (1.5 mL) was added to a solution of tert-butyl 5-[[6-[[2-(tert-butoxycarbonylamino)acetyl]amino]-5-fluoro-3-pyridinyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate (150 mg, 0.23 mmol) in DCM (2 mL), stirred at room temperature for 18 h, and concentrated under reduced pressure. The residue was triturated with diethyl ether (2 × 2 mL) and n-pentane (2 × 2 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).

[0430] M / z 376.24 (M+H) +

[0431] f. 5-[[5-Fluoro-6-[(2-guanidinoacetyl)amino]-3-pyridinyl]sulfonylamino]thiazole-4-carboxylic acid

[0432]

[0433] At room temperature, 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-pyridinyl]sulfonylamino]thiazole-4-carboxylic acid trifluoroacetate (120 mg, 0.32 mmol) in DMF (2 mL). The resulting reaction mixture was stirred at room temperature for 4 h, concentrated under reduced pressure and 1N ice HCl (2 mL) was added to the crude compound and stirred for 10 min. The resulting precipitate was filtered, washed with diethyl ether (2×5 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%).

[0434] 1 H NMR (400 MHz, DMSO-d 6 ) δ 13.20 (1H, brs), 10.8 (1H, brs), 8.51 (1H, d, J = 1.6 Hz), 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).

[0435] M / z 418.18 (M+H) +

[0436] LC-MS conditions:

[0437] Column: Acquity BEH C18 (50 mm x 2.1 mm, 1.7 um);

[0438] Mobile phase: A: aqueous solution of 0.05% formic acid; B: ACN solution of 0.05% formic acid;

[0439] Time (min) / % B: 0 / 3, 0.4 / 3, 3.2 / 98, 3.8 / 98, 4.2 / 3, 4.5 / 3;

[0440] Column temperature: 35 °C;

[0441] Flow rate: 0.6 mL / min.

[0442] Prep.HPLC conditions:

[0443] Column used: PHENYL HEXYL (150*30) mm 5u;

[0444] Mobile phase: (A) 0.1% formic acid, (B) acetonitrile;

[0445] Flow rate: 19 mL / min;

[0446] Gradient - (T / %B): 0 / 5, 1 / 5, 6 / 30, 8.9 / 30, 8.95 / 99, 11 / 99, 11.1 / 5, 14 / 5;

[0447] Solubility: ACN + THF.

[0448] Example 4

[0449] 5 - [[4 - [(3 - amino - 3 - imino - propionyl)amino] - 3 - fluoro - phenyl]sulfonylamino]thiazole - 4 - carboxylic acid

[0450]

[0451] a. N - [3 - amino - 1 - (tert - butoxycarbonylamino) - 3 - oxo - prop - 1 - enyl]carbamic acid tert - butyl ester

[0452]

[0453] At room temperature, saturated NaHCO 3 solution (10 mL) was added to a stirred solution of 3 - amino - 3 - imino - propionamide (3 g, 29.6 mmol) in dioxane (20 mL). Then (Boc) 2 O (16.5 mL, 74.0 mmol) was added dropwise at 0 °C. The resulting reaction mixture was stirred at room temperature for 16 h, concentrated under reduced pressure, and water (30 mL) was added to the residue. The crude compound was extracted with ethyl acetate (2 × 50 mL). The combined organic layers were dried over Na 2 SO 4 filtered and concentrated. The crude material was purified by flash column chromatography (eluting with a DCM solution of 2% methanol) to give an off - white solid (3.1 g, 34%).

[0454] M / z 302.36 (M + H) +

[0455] b. tert - butyl 5 - [[4 - [3,3 - bis(tert - butoxycarbonylamino)prop - 2 - enoyl]amino] - 3 - fluoro - phenyl]sulfonyl - [(4 - methoxyphenyl)methyl]amino]thiazole - 4 - carboxylate

[0456]

[0457] The solution of tert-butyl 5-[(4-bromo-3-fluorophenyl)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 minutes. Then, tert-butyl N-[3-amino-1-(tert-butoxycarbonylamino)-3-oxo-prop-1-enyl]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) were added under an argon atmosphere. The resulting reaction mixture was heated to 65 °C in a sealed tube, maintained for 3 hours, cooled to room temperature, filtered through a pad of diatomaceous earth, and the pad was washed with EtOAc (2 × 10 mL). The filtrate was concentrated under reduced pressure. The resulting crude compound was dissolved in ethyl acetate (50 mL) and washed with water (30 mL) and brine solution (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 a petroleum ether solution of 55% ethyl acetate) to give a pale yellow solid (1.3 g, 85%).

[0458] M / z 778.52 (M+H) +

[0459] c. 5-[[4-[(3-amino-3-imino-propionyl)amino]-3-fluorophenyl]sulfonylamino]thiazole-4-carboxylic acid

[0460]

[0461] At room temperature, TFA (6 mL) was added to tert-butyl 5-[[4-[3,3-bis(tert-butoxycarbonylamino)prop-2-enoyl]amino]-3-fluorophenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate (600 mg, 0.77 mmol). The resulting mixture was stirred for 3 hours and concentrated under reduced pressure. The resulting crude product was triturated with diethyl ether (2 × 10 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%).

[0462] 1 H NMR (400 MHz, DMSO-d 6)δ13.42(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).

[0463] M / z 402.3(M + H) +

[0464] LC-MS conditions:

[0465] Column: Acquity BEH C18(50mm x 2.1mm, 1.7um);

[0466] Mobile phase: A: Aqueous solution of 0.05% formic acid; B: ACN solution of 0.05% formic acid;

[0467] Time (min) / % B: 0 / 3, 0.4 / 3, 3.2 / 98, 3.8 / 98, 4.2 / 3, 4.5 / 3;

[0468] Column temperature: 35°C;

[0469] Flow rate: 0.6 mL / min.

[0470] Prep.HPLC conditions:

[0471] Column used: PRONTOSIL(250 * 19)mm, 5u;

[0472] Mobile phase: (A) 0.1% formic acid, (B) acetonitrile;

[0473] Flow rate: 19 mL / min;

[0474] Gradient - (T / % B): 0 / 5, 1 / 5, 7.3 / 59, 7.4 / 99, 11 / 99, 11.1 / 5, 14 / 5;

[0475] Solubility: ACN + THF + H2O + formic acid.

[0476] Examples 5 and 6

[0477] Example 5: 5 - [[3 - cyano - 4 - [(2 - guanidinoacetyl)amino]phenyl]sulfonylamino]thiazole - 4 - carboxylic acid

[0478]

[0479] Example 6: 5 - [[3 - carbamoyl - 4 - [(2 - guanidinoacetyl)amino]phenyl]sulfonylamino]thiazole - 4 - carboxylic acid

[0480]

[0481] a. 4-Bromo-3-cyano-benzenesulfonyl chloride

[0482]

[0483] Solution A: At 0 °C, concentrated HCl (5 mL) was added to a stirred solution of 5-amino-2-bromo-benzonitrile (2 g, 10.1 mmol) in AcOH (25 mL), and the mixture was stirred for 10 minutes. Then, a solution of NaNO 2 (770 mg, 11.1 mmol) in H 2 O (10 mL) was added and the mixture was stirred for 20 minutes.

[0484] Solution B: SO2 gas was bubbled into AcOH (25 mL) for 30 minutes. Then, CuCl 2 (1.62 g, 12.2 mmol) in H 2 O (10 mL) was added at 0 °C and the mixture was stirred for 20 minutes. Thereafter, Solution B was added dropwise to Solution A. The reaction mixture was stirred at room temperature for 20 minutes and diluted with water (20 mL). The resulting precipitate was filtered, washed with n-pentane (2 × 20 mL) and dried under high vacuum to obtain a yellow solid (1.7 g, 60%).

[0485] b. 5-[[3-Cyano-4-[(2-hydroxyacetyl)amino]phenyl]sulfonyl-methyl-amino]thiazole-4-carboxylic acid

[0486]

[0487] This compound was prepared according to the method reported in Step b of Example 2.

[0488] M / z 658.8 (M+H) +

[0489] c. 5-[[4-[(2-Aminoacetyl)amino]-3,5-difluoro-phenyl]sulfonylamino]thiazole-4-carboxylic acid trifluoroacetate

[0490]

[0491] This compound was prepared according to the method reported in Step c of Example 2.

[0492] M / z 382.4 (M+H) +

[0493] 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

[0494]

[0495] At room temperature, TFA:H 2 O (9:1, 5 mL) was added to tert-butyl 5-[[4-[[2-(tert-butoxycarbonylamino)acetyl]amino]-3-cyano-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate (400 mg, 0.60 mmol). The reaction mixture was stirred for 6 h and concentrated under reduced pressure. The resulting material was triturated with diethyl ether (2 × 10 mL) and dried under high vacuum to give a yellow solid which was used in the next step without further purification (300 mg, crude) (72% of 5-[[4-[(2-aminoacetyl)amino]-3-cyano-phenyl]sulfonylamino]thiazole-4-carboxylic acid and 8% of 5-[[4-[(2-aminoacetyl)amino]-3-carbamoyl-phenyl]sulfonylamino]thiazole-4-carboxylic acid).

[0496] M / z 382.05 (M+H) + and 400.01 (M+H) +

[0497] e. 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

[0498]

[0499] At room temperature, pyrazole-1-carboximidamide (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-4-carboxylic acid and 5-[[4-[(2-aminoacetyl)amino]-3-carbamoyl-phenyl]sulfonylamino]thiazole-4-carboxylic acid (300 mg, 0.78 mmol) in DMF (5 mL). The resulting reaction mixture was stirred at room temperature for 16 h and concentrated under reduced pressure. Water (5 mL) was added to the residue. The resulting precipitate was filtered and washed with diethyl ether (2 × 5 mL). The crude product was purified by preparative HPLC to give the title products:

[0500] Example 5

[0501] (72 mg, off-white solid):

[0502] 1 H 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).

[0503] M / z 424.34 (M+H) +

[0504] Example 6

[0505] (5.2 mg, off-white solid):

[0506] 1 H NMR (400 MHz, DMSO-d 6 ) 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).

[0507] M / z 442.34 (M+H) +

[0508] LC-MS conditions:

[0509] Column: Acquity BEH C18 (50 mm x 2.1 mm, 1.7 um);

[0510] Mobile phase: A: aqueous solution of 0.05% formic acid; B: ACN solution of 0.05% formic acid;

[0511] Time (min) / % B: 0 / 3, 0.4 / 3, 3.2 / 98, 3.8 / 98, 4.2 / 3, 4.5 / 3;

[0512] Column temperature: 35 °C;

[0513] Flow rate: 0.6 mL / min.

[0514] Prep.HPLC conditions:

[0515] Column: Symmetry C18(150*25)mm, 10u;

[0516] Mobile phase: (A) 0.05% formic acid (B) acetonitrile;

[0517] Flow rate: 19 mL / min;

[0518] Gradient (T / %B): 0 / 5, 1 / 5, 5 / 20, 10.5 / 24, 10.52 / 99, 12 / 99, 12.02 / 5, 15 / 5;

[0519] Solubility: ACN + H 2 O + THF + FA.

[0520] The following table shows compounds prepared using a method similar to that described in Examples 2 to 6 and purified in a similar manner by preparative HPLC.

[0521]

[0522]

[0523]

[0524]

[0525]

[0526]

[0527] Example 20

[0528] 5-[[4-[(2-Guanidinoacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylic acid

[0529]

[0530] a. Ethyl 5-[[4-[[2-(tert-Butoxycarbonylamino)acetyl]amino]phenyl]sulfonylamino]thiazole-4-carboxylate

[0531]

[0532] At room temperature, 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 room temperature for 15 minutes and then at the same temperature and N 2Ethyl 5-[(4-aminophenyl)sulfonylamino]thiazole-4-carboxylate (400 mg, 1.22 mmol) was added under an atmosphere. The resulting reaction mixture was stirred at room temperature for 16 h and concentrated under reduced pressure. The obtained crude compound was dissolved in a solution of DCM (20 mL) with 10% MeOH and washed with saturated NH 4 Cl (2 × 10 mL), water (10 mL), and brine solution (10 mL). The organic layer was dried over Na 2 SO 4 , filtered, and concentrated under vacuum. The crude material was purified by column chromatography (eluting with 3% MeOH) to give an off-white solid (400 mg, 67%).

[0533] M / z 484.8 (M+H) + 507.06 (M+Na) +

[0534] b. Ethyl 5-[[4-[(2-aminoacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylate

[0535]

[0536] A solution of 2N HCl in Et 2 O (4 mL) was added to a solution of 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 at room temperature for 5 h and concentrated under reduced pressure. The crude product was purified by preparative HPLC (HCOOH / CH 3 CN / H 2 O) to give an off-white solid (300 mg, 94%).

[0537] M / z 385.13 (M+H) +

[0538] c. Ethyl 5-[[4-[[2-[[N,N'-bis(tert-butoxycarbonyl)formamidinyl]amino]acetyl]amino]phenyl]sulfonylamino]thiazole-4-carboxylate

[0539]

[0540] At room temperature, DIPEA (0.08 mL, 0.49 mmol) and tert-butyl N-[(tert-butoxycarbonylamino)-pyrazol-1-yl-methylene]carbamate (87 mg, 0.28 mmol) were added to a stirred solution of ethyl 5-[[4-[(2-aminoacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylate (270 mg, 0.70 mmol) in DMF (5 mL). The resulting reaction mixture was stirred at room temperature for 16 h and concentrated under reduced pressure. The obtained crude compound was dissolved in a solution of 10% MeOH in DCM (20 mL) and washed with water (10 mL) and brine solution (10 mL). The organic layer was dried with Na 2 SO 4 , filtered and concentrated under reduced pressure. The crude material was purified by column chromatography (eluting with a solution of 4% MeOH in DCM) to give an off-white solid (250 mg, 56%).

[0541] M / z 626.97 (M+H) +

[0542] d. Ethyl 5-[[4-[[2-[[(Z)-N,N'-bis(tert-butoxycarbonyl)formamidinyl]amino]acetyl]amino]phenyl]sulfonylamino]thiazole-4-carboxylate

[0543]

[0544] At room temperature and under N 2 atmosphere, TMSOK (69 mg, 0.54 mmol) was added to a stirred solution of ethyl 5-[[4-[[2-[[N,N'-bis(tert-butoxycarbonyl)formamidinyl]amino]acetyl]amino]phenyl]sulfonylamino]thiazole-4-carboxylate (170 mg, 0.27 mmol) in THF (4 mL). The resulting reaction mixture was stirred at 40 °C for 5 h and concentrated under reduced pressure. The obtained crude product was triturated with diethyl ether (2 × 5 mL) and dried under high vacuum. The residue was dissolved in water (5 mL) and acidified with 1 N HCl (adjusting pH~2). The resulting solid was filtered, washed with n-pentane and dried under high vacuum to give an off-white solid which was used in the next step without further purification (70 mg crude, 43%).

[0545] M / z 598.92 (M+H) +

[0546] e. 5-[[4-[(2-Guanidinoacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylic acid

[0547]

[0548] At 0 °C, a solution of 2N HCl in diethyl ether (1 mL) was added to a solution of 5-[[4-[[2-[[(Z)-N,N'-bis(tert-butoxycarbonyl)formamidinyl]amino]acetyl]amino]phenyl]sulfonylamino]thiazole-4-carboxylic acid (70 mg, 0.11 mmol) in diethyl ether (2 mL). The reaction mixture was stirred at room temperature 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%).

[0549] 1 H NMR (500 MHz, DMSO-d 6 ) 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).

[0550] M / z 398.78 (M + H) +

[0551] Example 21

[0552] 5-[[4-[[2-(4,5-Dihydro-1H-imidazol-2-yl)acetyl]amino]-3-fluorophenyl]sulfonylamino]thiazole-4-carboxylic acid

[0553]

[0554] a. tert-Butyl 5-[[4-[(2-cyanoacetyl)amino]-3-fluorophenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

[0555]

[0556] 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 min. The temperature of the reaction mixture was cooled to room temperature, and a solution of tert-butyl 5-[(4-amino-3-fluorophenyl) sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate (500 mg,

[0557] 1.01 mmol) in DCM (30 mL) was added under a nitrogen atmosphere. The resulting reaction mixture was heated to reflux for 2.5 h, cooled to room temperature, diluted with DCM (50 mL), and washed with aqueous NaHCO 3 solution (30 mL), water (30 mL), and brine (30 mL). The organic layer was dried over Na 2 SO 4Dry, filter and concentrate under reduced pressure. Purify the crude material by flash chromatography (eluting with a DCM solution of 1% to 2% MeOH) to afford a light yellow solid (180 mg, 31%).

[0558] M / z 561.43 (M+H) +

[0559] b. tert-Butyl 5-[[4-[[2-(4,5-dihydro-1H-imidazol-2-yl)acetyl]amino]-3-fluorophenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

[0560]

[0561] At 0 °C, HCl gas was bubbled 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 ethanol:Et 2 O (1:2, 30 mL) for 2 h. The resulting reaction mixture was kept in the refrigerator for 16 h. Then the volatile components were evaporated under reduced pressure at 40 °C. The residue was dissolved in ethanol (10 mL) and ethylenediamine (48 mg, 0.80 mmol) was added at room temperature. The reaction mixture was stirred at room temperature for 16 h and concentrated under reduced pressure. The obtained crude product was triturated with diethyl ether (2 × 5 mL) and dried under high vacuum to give a light brown solid, which was used for the next step without further purification (330 mg, crude).

[0562] M / z 548.29 (M-Boc+H) +

[0563] c. 5-[[4-[[2-(4,5-dihydro-1H-imidazol-2-yl)acetyl]amino]-3-fluorophenyl]sulfonylamino]thiazole-4-carboxylic acid

[0564]

[0565] At room temperature, 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). The reaction mixture was stirred at room temperature for 4 h and concentrated under reduced pressure. The obtained crude product was triturated with diethyl ether (2 × 5 mL) and dried under high vacuum. Purify the crude product by preparative HPLC to afford the title product as an off-white solid (26 mg, 11%).

[0566] 1 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), 7.54 - 7.48 (2H, m), 3.40 (2H, s), 3.36 - 3.28 (2H, obs), 2.88 - 2.85 (2H, m).

[0567] M / z 428.37 (M + H) +

[0568] Example 22

[0569] 5 - [[3 - fluoro - 4 - [[3 - imino - 3 - (methylamino) propionyl] amino] phenyl] sulfonylamino] thiazole - 4 - carboxylic acid

[0570]

[0571] a. tert - Butyl 5 - [[3 - fluoro - 4 - [[3 - [hydroxy(methyl) amino] - 3 - imino - propionyl] amino] phenyl] sulfonyl - [(4 - methoxyphenyl) methyl] amino] thiazole - 4 - carboxylate

[0572]

[0573] At room temperature, 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). The resulting reaction mixture was stirred at 60 °C for 3 h, cooled to room temperature, filtered and washed with ethanol (2 × 10 mL). The combined organic layers were concentrated under reduced pressure. The obtained crude compound was triturated with Et 2 O (2 × 10 mL) and dried under high vacuum to give a brown solid, which was used in the next step without further purification.

[0574] M / z 608.03 (M + H) +

[0575] b. tert - Butyl 5 - [[3 - fluoro - 4 - [[3 - imino - 3 - (methylamino) propionyl] amino] phenyl] sulfonyl - [(4 - methoxyphenyl) methyl] amino] thiazole - 4 - carboxylate

[0576]

[0577] At room temperature, bis(pinacolato)diboron (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-propionyl]amino]phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate (570 mg, 0.93 mmol) in acetonitrile (10 mL). The resulting reaction mixture was stirred at room temperature for 1 h and concentrated under reduced pressure. The crude compound was purified by flash chromatography (eluting with a 10% methanol and DCM solution containing 2% triethylamine) to afford a pale yellow solid (130 mg, 23%).

[0578] M / z 592.05 (M + H) +

[0579] c. 5-[[3-Fluoro-4-[[3-imino-3-(methylamino)propionyl]amino]phenyl]sulfonylamino]thiazole-4-carboxylic acid

[0580]

[0581] At room temperature, TFA (3 mL) was added to tert-butyl 5-[[3-fluoro-4-[[3-imino-3-(methylamino)propionyl]amino]phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate (130 mg, 0.21 mmol). The reaction mixture was stirred at room temperature for 2 h and concentrated under reduced pressure. The resulting crude product was triturated with diethyl ether (2 × 5 mL) and dried under high vacuum. The crude product was purified by preparative HPLC to afford the title product as a yellow solid (20 mg, 22%).

[0582] 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).

[0583] M / z 416.34 (M + H) +

[0584] LC-MS conditions:

[0585] Column: Acquity BEH C18 (50 mm x 2.1 mm, 1.7 um);

[0586] Mobile phase: A: Aqueous solution of 0.05% formic acid; B: ACN solution of 0.05% formic acid;

[0587] Time (min) / % B: 0 / 3, 0.4 / 3, 3.2 / 98, 3.8 / 98, 4.2 / 3, 4.5 / 3;

[0588] Column temperature: 35 °C;

[0589] Flow rate: 0.6 mL / min.

[0590] Prep.HPLC conditions:

[0591] Column: Symmetry C18 (300*19) mm, 7u;

[0592] Mobile phase: (A) 0.1% formic acid (B) acetonitrile;

[0593] Flow rate: 19 mL / min;

[0594] Gradient (T / % B): 0 / 5, 1 / 5, 8.9 / 40, 8.92 / 99, 12 / 99, 12.1 / 5, 15 / 5;

[0595] Solubility: ACN + H 2 O + THF.

[0596] Example 23

[0597] 5-[[2-[(2-Guanidinoacetyl)amino]thiazol-5-yl]sulfonylamino]thiazole-4-carboxylic acid

[0598]

[0599] a. 2-Acetamidothiazole-5-sulfonyl chloride

[0600]

[0601] At 0 °C, N-thiazol-2-ylacetamide (5 g, 35.2 mmol) was added portionwise to chlorosulfonic acid solution (11.7 mL, 176 mmol). The reaction mixture was stirred at 100 °C for 4 h, cooled to room temperature and poured into ice water (100 mL). The resulting precipitate was filtered and washed with water (20 mL). The precipitate was triturated with n-pentane (2×20 mL) and azeotroped with toluene to give an off-white solid which was used in the next step without further purification (2 g crude, 23%).

[0602] M / z 241.23 (M+H) +

[0603] b. Ethyl 5-[(2-acetamidothiazol-5-yl)sulfonylamino]thiazole-4-carboxylate

[0604]

[0605] At 0 °C, 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), and the mixture was stirred for 5 minutes. Then, at 0 °C, a solution of 2-acetamidothiazole-5-sulfonyl chloride (502 mg, 2.0 mmol) in THF (10 mL) was added to the reaction mixture. The reaction mixture was stirred at the same temperature for 1 hour. Ice water (30 mL) was added to the reaction mixture, and then it was washed with EtOAc (2 × 15 mL). The aqueous layer was acidified to pH 2.0 with 1N HCl and extracted with EtOAc (3 × 15 mL). The organic layer was dried over Na 2 SO 4 dried, filtered, and concentrated under reduced pressure to give a light brown solid, which was used in the next step without further purification (175 mg crude, 26%).

[0606] M / z 377.32 (M+H) +

[0607] c. Ethyl 5-[(2-aminothiazol-5-yl)sulfonylamino]thiazole-4-carboxylate hydrochloride

[0608]

[0609] At room temperature, concentrated HCl (7 mL) was added to a solution of ethyl 5-[(2-acetamidothiazol-5-yl)sulfonylamino]thiazole-4-carboxylate (700 mg, 1.86 mmol) in ethanol (70 mL). The reaction mixture was refluxed for 5 hours and concentrated under reduced pressure. The resulting crude compound was washed with diethyl ether (20 mL) and 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).

[0610] M / z 335.04 (M+H) +

[0611] d. Ethyl 5-[[2-[[2-(tert-butoxycarbonylamino)acetyl]amino]thiazol-5-yl]sulfonylamino]thiazole-4-carboxylate

[0612]

[0613] At room temperature, 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). The reaction mixture was stirred at room temperature for 15 minutes and then ethyl 5-[(2-aminothiazol-5-yl)sulfonylamino]thiazole-4-carboxylate hydrochloride (600 mg, 1.79 mmol) was added under the same temperature and N 2 atmosphere. The resulting reaction mixture was stirred at room temperature for 18 hours. Ice water (30 mL) was added and the mixture was extracted with DCM (3 × 20 mL). The organic layer was dried over Na 2 SO 4 4, filtered and concentrated under reduced pressure. The crude compound was purified by flash chromatography (eluting with a petroleum ether solution of 60% to 80% EtOAc) to give a brown solid (400 mg, 45%).

[0614] M / z 492.34 (M+H) +

[0615] e. 5-[[2-[[2-(tert-Butoxycarbonylamino)acetyl]amino]thiazol-5-yl]sulfonylamino]thiazole-4-carboxylic acid

[0616]

[0617] At room temperature, 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). The reaction mixture was stirred at 40 °C for 1 hour, concentrated under reduced pressure and water (2 mL) was added to the residue. The reaction mixture was acidified to pH 2 with 1N HCl. The resulting precipitate was filtered, washed with diethyl ether (2 × 10 mL) and n-pentane (10 mL) and dried under high vacuum to give a pale yellow solid (200 mg, 53%).

[0618] M / z 464.30 (M+H) +

[0619] f. 5-[[2-[(2-Aminoacetyl)amino]thiazol-5-yl]sulfonylamino]thiazole-4-carboxylic acid hydrochloride

[0620]

[0621] At room temperature, Et of HCl 2The O solution (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). 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 (2 × 5 mL) and n-pentane (5 mL) to give a pale yellow solid, which was used in the next step without further purification (150 mg, crude).

[0622] M / z 364.30 (M+H) +

[0623] g. 5-[[2-[(2-guanidinoacetyl)amino]thiazol-5-yl]sulfonylamino]thiazole-4-carboxylic acid

[0624]

[0625] At room temperature, DIPEA (0.44 mL, 2.7 mmol) was added to a stirred solution of 5-[[2-[(2-aminoacetyl)amino]thiazol-5-yl]sulfonylamino]thiazole-4-carboxylic acid hydrochloride (100 mg, 0.27), pyrazole-1-carboxamidine hydrochloride (80 mg, 0.55 mmol) in DMF (2 mL). The reaction mixture was stirred at the same temperature for 6 hours. The DMF was evaporated, then water (3 mL) was added to the resulting crude material and stirred for 5 minutes. The resulting precipitate was filtered and washed with water (2 × 2 mL), then dried under high vacuum. The crude compound was purified by preparative HPLC to give an off-white solid (16 mg, 14%).

[0626] 1 H NMR (400 MHz, DMSO-d 6 ) 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).

[0627] M / z 405.9 (M+H) +

[0628] LC-MS conditions:

[0629] Column: Acquity BEH C18 (50 mm x 2.1 mm, 1.7 um)

[0630] Mobile phase: A: aqueous solution of 0.05% formic acid; B: ACN solution of 0.05% formic acid

[0631] Gradient: Time (min) / %B: 0 / 3, 0.4 / 3, 3.2 / 98, 3.8 / 98, 4.2 / 3, 4.5 / 3

[0632] Column temperature: 35 °C,

[0633] Flow rate: 0.6 mL / min

[0634] Prep.HPLC conditions:

[0635] Column used: Atlantis T3 (250*19) mm, 5u;

[0636] Mobile phase: (A) 0.1% formic acid (B) acetonitrile

[0637] Flow rate: 19 mL / min

[0638] Gradient - (T / %B): 0 / 5, 1 / 5, 8.2 / 55, 8.21 / 99, 10 / 99, 10.1 / 5, 13 / 5

[0639] Diluent: ACN + H 2 O + FA

[0640] Example 24

[0641] 5 - [[4 - [2 - (2 - formamidinohydrazino)-2 - oxo - ethyl]-3 - fluoro - phenyl]sulfonylamino]thiazole - 4 - carboxylic acid

[0642]

[0643] a. tert - Butyl 5 - [[4 - (2 - ethoxy - 2 - oxo - ethyl)-3 - fluoro - phenyl]sulfonyl - [(4 - methoxyphenyl)methyl]amino]thiazole - 4 - carboxylate

[0644]

[0645] Purge a mixture of tert - butyl 5 - [(4 - bromo - 3 - fluoro - phenyl) sulfonyl - [(4 - methoxyphenyl)methyl]amino]thiazole - 4 - carboxylate (2 g, 3.58 mmol), potassium 3 - ethoxy - 3 - oxo - propionate (1.2 g, 7.16 mmol) and DMAP (43 mg, 0.35 mmol) in mesitylene (20 mL) with argon for 30 minutes. Then add BINAP (222 mg, 0.35 mmol) and Pd 2 (dba) 3(327 mg, 0.35 mmol). The reaction mixture was stirred at 120 °C for 18 h, cooled to room temperature and concentrated under reduced pressure. The crude product was purified by flash chromatography (eluting with a petroleum ether solution of 40% EtOAc) to give a yellow solid which was used in the next step without further purification (0.45 g, crude).

[0646] M / z 565.43 (M+H)+

[0647] b. tert-Butyl 5-[[3-fluoro-4-(2-hydrazino-2-oxo-ethyl)phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

[0648]

[0649] At room temperature, hydrazine hydrate (709 mg, 14.1 mmol) was added to a stirred solution of tert-butyl 5-[[4-(2-ethoxy-2-oxo-ethyl)-3-fluorophenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate (400 mg, 0.7 mmol) in ethanol (20 mL). 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).

[0650] M / z 551.42 (M+H) +

[0651] c. tert-Butyl 5-[[4-[2-[2-N,N'-bis(tert-butoxycarbonyl)formimidoyl]hydrazino]-2-oxo-ethyl]-3-fluorophenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

[0652]

[0653] At room temperature, 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 N-N-[(tert-butoxycarbonylamino)-pyrazol-1-yl-methylene]carbamate (394 mg, 1.27 mmol) in DMF (5 mL). The reaction mixture was stirred at the same temperature for 18 h. Ice water was added to the reaction mixture and stirred for 10 min. The resulting precipitate was filtered, washed with water (2×5 mL) and dried under high vacuum. The crude product was purified by flash chromatography (eluting with a petroleum ether solution of 60% EtOAc) to give a yellow solid (120 mg, 23%).

[0654] M / z 793.53 (M+H) +

[0655] d. 5-[[4-[2-(2-Carbamimidoylhydrazino)-2-oxo-ethyl]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylic acid

[0656]

[0657] At room temperature, 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). The reaction mixture was stirred at the same temperature for 3 h and concentrated under reduced pressure. The resulting crude material was triturated with diethyl ether (2×5 mL). The crude product was purified by preparative HPLC to give the title product (23 mg) as an off-white solid.

[0658] 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).

[0659] M / z 417.35 (M+H) +

[0660] LC-MS conditions:

[0661] Column: Acquity BEH C18 (50 mm x 2.1 mm, 1.7 um);

[0662] Mobile phase: A: Aqueous solution of 0.05% formic acid; B: ACN solution of 0.05% formic acid;

[0663] Time (min) / % B: 0 / 3, 0.4 / 3, 3.2 / 98, 3.8 / 98, 4.2 / 3, 4.5 / 3;

[0664] Column temperature: 35 °C;

[0665] Flow rate: 0.6 mL / min.

[0666] Prep.HPLC conditions:

[0667] Column used: Symmetry C18 (300×19) mm, 7u;

[0668] Mobile phase: (A) 0.05% formic acid (B) acetonitrile;

[0669] Flow rate: 19 mL / min;

[0670] Gradient (T / %B): 0 / 2, 1 / 2, 0, 8 / 20, 10.5 / 20, 10.51 / 99, 12 / 99, 12.1 / 2, 15 / 2;

[0671] Solubility: ACN + H2O + THF.

[0672] The compounds prepared by a method similar to that described in Examples 20 to 24 above and purified in a similar manner by preparative HPLC are shown in the following table:

[0673]

[0674] Example 26

[0675] 5-[[3,5-Difluoro-4-(guanidinoaminocarbonylamino)phenyl]sulfonylamino]thiazole-4-carboxylic acid

[0676]

[0677] a. tert-Butyl 5-[(4-amino-3,5-difluorophenyl)sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

[0678]

[0679] Add a saturated solution of NH 3 in dioxane (120 mL) to a mixture of tert-butyl 5-[(4-bromo-3,5-difluorophenyl)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). At 100 °C, stir the resulting mixture in a sealed tube for 5 hours, filter through a pad of diatomaceous earth, and wash the pad with ethyl acetate (2 × 25 mL). Concentrate the filtrate under reduced pressure. Purify the crude material by flash chromatography (eluting with a 50% ethyl acetate in petroleum ether solution) to give a pale yellow solid (1.25 g, 70%).

[0680] M / z 512.4 (M+H) + ; 534.56 (M+Na) +

[0681] b. tert-Butyl 5-[[3,5-difluoro-4-[(4-nitrophenoxy)carbonylamino]phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

[0682]

[0683] At room temperature, (4-nitrophenyl) chloroformate (1.57 g, 7.82 mmol) was added to a stirred solution of tert-butyl 5-[(4-amino-3,5-difluorophenyl)sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate (2 g, 3.91 mmol) in toluene (120 mL), and the mixture was 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).

[0684] c. tert-Butyl 5-[[4-[(tert-butoxycarbonylamino)carbamoylamino]-3,5-difluorophenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

[0685]

[0686] At 0 °C, DIPEA (2.6 mL, 15.5 mmol) was added to a 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). The reaction mixture was stirred at room temperature for 3 hours and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with a petroleum ether solution of 70% ethyl acetate) to give a pale yellow solid (1.5 g, 43%).

[0687] M / z 670.4 (M+H) +

[0688] d. Hydrochloride of 5-[[3,5-difluoro-4-(hydrazinecarbonylamino)phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylic acid

[0689]

[0690] At room temperature, Et of HCl 2The O solution (2 M, 200 mL) was added to tert-butyl 5-[[4-[(tert-butoxycarbonylamino)carbamoyl amino]-3,5-difluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate (1.5 g, 2.24 mmol). The reaction mixture was stirred for 24 h, cooled to 0 °C for 30 min and the Et 2 O was decanted. The crude product was triturated with diethyl ether (2 × 40 mL) and dried under high vacuum to give an off-white solid (1 g, crude).

[0691] M / z 514.3 (M+H) +

[0692] e. 5-[[4-[[[N,N'-Bis(tert-butoxycarbonyl)formamidinyl]amino]carbamoyl amino]-3,5-difluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylic acid

[0693]

[0694] At room temperature, 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 acid hydrochloride (1 g, 1.75 mmol) and tert-butyl (NZ)-N-[(tert-butoxycarbonylamino)-pyrazol-1-yl-methylene]carbamate (0.54 g, 1.75 mmol) in DMF (6 mL). The reaction mixture was stirred for 5 h and the DMF was removed. Then water was added to the crude product and stirred for 5 min. The resulting precipitate was filtered, washed with water (2 × 5 mL) and dried under high vacuum to give an off-white solid (1.25 g, crude).

[0695] M / z 756.1 (M+H) +

[0696] f. 5-[[3,5-Difluoro-4-(guanidinocarbamoyl amino)phenyl]sulfonylamino]thiazole-4-carboxylic acid

[0697]

[0698] At room temperature, TFA (13 mL) was added to 5-[[4-[[[(Z)-N,N'-bis(tert-butoxycarbonyl)formamidinyl]amino]carbamoyl amino]-3,5-difluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylic acid (1.25 g, 1.54 mmol). The reaction mixture was stirred at room temperature for 3 h and concentrated by N 2The crude product was evaporated with TFA by gas purge. The obtained crude product was triturated with diethyl ether and purified by preparative HPLC to give the title compound (150 mg) as a white solid.

[0699] 1 1H 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).

[0700] M / z 436.0 (M+H) +

[0701] LC-MS conditions:

[0702] Column: Aquity UPLC BEH C18 (50 mm x 2.1 mm, 1.7 um);

[0703] Mobile phase: A: aqueous solution of 0.1% formic acid, B: acetonitrile solution of 0.1% formic acid;

[0704] Gradient: time (min) / % B 0 / 2, 0.2 / 2, 1.5 / 98, 2.6 / 98, 2.61 / 2, 3.2 / 2;

[0705] Column temperature: 45 °C, flow rate: 0.8 mL / min

[0706] Prep.HPLC conditions:

[0707] Column: X select C18 (150 * 30 mm), 5u;

[0708] Mobile phase: H 2 O solution of 0.05% formic acid: acetonitrile;

[0709] Flow rate: 25 mL / min;

[0710] Gradient (T / % B): 0 / 50, 8 / 50, 8 / 40, 9 / 40, 9.1 / 98, 11 / 98, 11.1 / 5, 14 / 40

[0711] Diluent: ACN + H 2 O + MeOH + THF.

[0712] Example 27

[0713] 5-[[3-Fluoro-4-(2-guanidinoethoxycarbonylamino)phenyl]sulfonylamino]thiazole-4-carboxylic acid

[0714]

[0715] a. tert-Butyl 5-[[3-fluoro-4-[(4-nitrophenoxy)carbonylamino]phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

[0716]

[0717] At room temperature, p-nitrophenyl chloroformate (1.33 g, 6.0 mmol) was added to a stirred solution of tert-butyl 5-[(4-amino-3-fluorophenyl)sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate (1 g, 2.02 mmol) in toluene (30 mL). The resulting reaction mixture was stirred at 120 °C for 1 h and concentrated under reduced pressure. The obtained crude product was triturated with n-pentane (2 × 10 mL) and dried under high vacuum to give an off-white solid, which was used in the next step without further purification (1.5 g, crude).

[0718] M / z 659.43 (M+H) +

[0719] b. tert-Butyl 5-[[4-[2-(tert-butoxycarbonylamino)ethoxycarbonylamino]-3-fluorophenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

[0720]

[0721] At room temperature, 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). The resulting reaction mixture was stirred at room temperature for 2 h. Ice water was added, and then the mixture was extracted with ethyl acetate (2 × 50 mL). The combined organic layers were dried over anhydrous Na 2 SO 4 , filtered and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with a petroleum ether solution of 40% ethyl acetate) to give an off-white solid (500 mg, 40%).

[0722] M / z 681.50 (M+H) +

[0723] c. Trifluoroacetate of 5-[[4-(2-aminoethoxycarbonylamino)-3-fluorophenyl]sulfonylamino]thiazole-4-carboxylic acid

[0724]

[0725] At room temperature, TFA (5 mL) was added to tert-butyl 5-[[4-[2-(tert-butoxycarbonylamino)ethoxycarbonylamino]-3-fluorophenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate (450 mg, 0.66 mmol). The reaction mixture was stirred at the same temperature for 24 h and concentrated under reduced pressure. The resulting crude product was triturated with diethyl ether (2 × 10 mL) and dried under high vacuum to give an off-white solid, which was used in the next step without further purification (350 mg, crude).

[0726] M / z 405.36 (M+H) +

[0727] d. 5-[[3-Fluoro-4-(2-guanidinoethoxycarbonylamino)phenyl]sulfonylamino]thiazole-4-carboxylic acid

[0728]

[0729] At room temperature, pyrazole-1-carboximidamide 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-fluorophenyl]sulfonylamino]thiazole-4-carboxylic acid trifluoroacetate (250 mg, 0.61 mmol) in DMF (6 mL). The resulting reaction mixture was stirred at room temperature for 4 h and concentrated under reduced pressure. Water (5 mL) was added to the residue. The resulting precipitate was filtered, washed with diethyl ether (2 × 5 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%).

[0730] 1 H 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), 7.62 - 7.56 (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).

[0731] M / z 447.27 (M+H) +

[0732] The compounds prepared using a method similar to that described in Examples 26 and 27 and purified in a similar manner by preparative HPLC are shown in the following table:

[0733]

[0734]

[0735] Example 32

[0736] 5-[[3,5-Difluoro-4-(guanidinoaminocarbonylamino)phenyl]sulfonylamino]thiazole-4-carboxylic acid

[0737]

[0738] a. tert-Butyl 5-[[4-[(2-chloroacetyl)amino]-3-fluorophenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

[0739]

[0740] At 0 °C, 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-fluorophenyl) sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate (450 mg, 0.91 mmol) in DCM (5 mL). The resulting reaction mixture was stirred at room temperature for 2 h, quenched with ice water (5 mL) and extracted with DCM (2 × 10 mL). The combined organic layers were dried over Na 2 SO 4 dried, filtered and concentrated under reduced pressure. The crude material was purified by trituration with diethyl ether (2 × 5 mL) to give a green solid which was used in the next step without further purification (400 mg, crude).

[0741] M / z 570.69 (M+H) +

[0742] b. tert-Butyl 5-[[4-[(2-azidoacetyl)amino]-3-fluorophenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

[0743]

[0744] At room temperature, NaN 3(92 mg, 1.40 mmol) was added to a stirred solution of tert-butyl 5-[[4-[(2-chloroacetyl)amino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]amino]thiazole-4-carboxylate (400 mg, 0.70 mmol) in DMF (5 mL). The resulting reaction mixture was stirred at room temperature for 16 h, quenched with ice water (10 mL) and extracted with ethyl acetate (2 × 10 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 (2 × 5 mL) to afford a light brown solid (370 mg, 91%).

[0745] M / z 577.23 (M+H) +

[0746] c. tert-butyl 5-[[4-[(2-aminoacetyl)amino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

[0747]

[0748] Under room temperature and nitrogen atmosphere, 10% Pd / C (300 mg) was added to a solution of tert-butyl 5-[[4-[(2-azidoacetyl)amino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate (370 mg, 0.64 mmol) in EtOAc (10 mL). The resulting reaction mixture was stirred at room temperature under a hydrogen atmosphere (balloon pressure) for 16 h, filtered through a pad of Celite and washed with EtOAc (20 mL). The filtrate was concentrated under reduced pressure. The crude material was purified by trituration with diethyl ether (2 × 5 mL) to afford a brown solid (340 mg, 96%).

[0749] M / z 551.35 (M+H) +

[0750] 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

[0751]

[0752] At room temperature, 2-methylthio-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). The resulting reaction mixture was heated to 70 °C in a sealed vial and maintained for 48 h, concentrated under reduced pressure to give a yellow solid which was used in the next step without further purification (500 mg, crude).

[0753] M / z 619.36 (M+H) +

[0754] e. 5-[[3,5-Difluoro-4-(guanidinoaminocarbonylamino)phenyl]sulfonylamino]thiazole-4-carboxylic acid

[0755]

[0756] At 0 °C, TFA (5 mL) was added to 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 (500 mg, 0.50 mmol), and the mixture was stirred at room temperature for 6 h. TFA was evaporated under reduced pressure, and the resulting crude product was triturated with diethyl ether (2 × 5 mL) and dried in vacuo. The crude product was purified by preparative HPLC to give the title product as a white solid (37 mg, 18%).

[0757] 1 H 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), 7.55 - 7.48 (2H, m), 4.08 (2H, s), 3.60 (4H, s).

[0758] M / z 443.24 (M+H) +

[0759] LC-MS conditions:

[0760] Column: Acquity BEH C18 (50 mm x 2.1 mm, 1.7 um);

[0761] Mobile phase: A: aqueous solution of 0.05% formic acid; B: ACN solution of 0.05% formic acid;

[0762] Time (min) / % B: 0 / 3, 0.4 / 3, 2 / 98, 3.8 / 98, 4.2 / 3, 4.5 / 3;

[0763] Column temperature: 35 °C,

[0764] Flow rate: 0.6 mL / min.

[0765] Prep.HPLC conditions:

[0766] Column: Atlantis T3 (250*19) mm, 5u;

[0767] Mobile phase: (A) 0.1% formic acid (B) acetonitrile;

[0768] Flow rate: 19 mL / min;

[0769] Gradient (T / % B): 0 / 5, 1 / 5, 9 / 30, 10.31 / 99, 12 / 99, 12.1 / 5, 15 / 5;

[0770] Solubility: ACN + H 2 O + THF.

[0771] Example 33

[0772] 5-[[3-Fluoro-4-[[2-(morpholine-4-carboxamidoamino)acetyl]amino]phenyl]sulfonylamino]thiazole-4-carboxylic acid

[0773]

[0774] a. tert-Butyl 5-[[3-fluoro-4-[[2-(morpholine-4-thiocarbonylamino)amino]acetyl]amino]phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

[0775]

[0776] 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 bis(imidazol-1-yl)methanethione (242 mg, 1.36 mmol) in CH 2 Cl 2 (15 mL) was stirred at room temperature for 30 minutes. Then morpholine (118 mg, 1.36 mmol) was added and the resulting reaction mixture was stirred at 40 °C for 1 hour. The reaction mixture was concentrated in vacuo. The crude compound was purified by flash chromatography, using CH 2 Cl 2Eluted with the solution to obtain a light pink gummy substance (100 mg, 83%).

[0777] M / z 680.42 (M+H) +

[0778] b. tert-Butyl 5-[[3-fluoro-4-[[2-(morpholine-4-carboxamido)amino]acetyl]amino]phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

[0779]

[0780] Ag(OTf) (255 mg, 0.99 mmol) was added to a solution of tert-butyl 5-[[3-fluoro-4-[[2-(morpholine-4-thiocarbonylamino)amino]acetyl]amino]phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate (450 mg, 0.66 mmol) in CH 2 Cl 2 :THF (1:1, 20 mL). The reaction mixture was cooled to -30 °C and purged with NH 3 gas for 15 minutes. The reaction mixture was stirred at room temperature for 4 hours, quenched with MeOH (1 mL) and concentrated under reduced pressure. Water (25 mL) was added and the mixture was extracted with EtOAc (2 × 50 mL). The combined organic layers were dried over Na 2 SO 4 and filtered and concentrated. The crude material was purified by flash chromatography, eluting with a 3% MeOH in CH 2 Cl 2 solution to obtain a black gummy substance (250 mg, 57%).

[0781] M / z 663.53 (M+H) +

[0782] c. 5-[[3-Fluoro-4-[[2-(morpholine-4-carboxamidoamino)acetyl]amino]phenyl]sulfonylamino]thiazole-4-carboxylic acid

[0783]

[0784] TFA:H of tert-butyl 5-[[3-fluoro-4-[[2-(morpholine-4-carboxamido)amino]acetyl]amino]phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate 2The O(95:5, 2 mL) solution was stirred at room temperature for 3 h. The reaction mixture was concentrated, and the crude product was neutralized with methanolic ammonia. Then the reaction mixture was concentrated under reduced pressure. The crude material was purified by preparative HPLC to give the title product as an off-white solid (12.4 mg, 8%).

[0785] 1 H NMR (400 MHz, DMSO-d 6 ) 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).

[0786] M / z 487.34 (M + H) +

[0787] LC-MS conditions:

[0788] Column: Acquity BEH C18 (50 mm x 2.1 mm, 1.7 um);

[0789] Mobile phase: A: aqueous solution of 0.05% formic acid; B: ACN solution of 0.05% formic acid;

[0790] Time (min) / % B: 0 / 3, 0.4 / 3, 3.2 / 98, 3.8 / 98, 4.2 / 3, 4.5 / 3;

[0791] Column temperature: 35 °C,

[0792] Flow rate: 0.6 mL / min.

[0793] Prep.HPLC conditions:

[0794] Column used: Symmetry C18 (300 × 19) mm, 7u;

[0795] Mobile phase: (A) 0.1% formic acid (B) acetonitrile;

[0796] Flow rate: 19 mL / min;

[0797] Gradient - (T / % B): 0 / 5, 1 / 5, 7.1 / 56, 7.15 / 99, 10 / 99, 10.1 / 5, 13 / 5;

[0798] Solubility: CH 3 CN + H 2O.

[0799] Example 34

[0800] 5-[[4-[[2-[(N-cyanoguanidino)amino]acetyl]amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylic acid

[0801]

[0802] At room temperature and under a nitrogen atmosphere, 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]acetyl]amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylic acid (200 mg, 0.53 mmol) in DMF (5 mL). The reaction mixture was stirred at 50 °C for 48 hours and concentrated under reduced pressure. The crude compound was diluted with water (5 mL) and acidified to pH ~2 to 3 with 1N HCl. The resulting precipitate was filtered 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%).

[0803] 1 H 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).

[0804] M / z 442.18 (M + H) +

[0805] LC-MS conditions:

[0806] Column: Acquity BEH C18 (50 mm x 2.1 mm, 1.7 um);

[0807] Mobile phase: A: aqueous solution of 0.05% formic acid; B: ACN solution of 0.05% formic acid;

[0808] Time (min) / % B: 0 / 3, 0.4 / 3, 2 / 98, 3.4 / 98, 3.5 / 3, 4 / 3;

[0809] Column temperature: 35 °C,

[0810] Flow rate: 0.6 mL / min.

[0811] Prep.HPLC conditions:

[0812] Column used: XBRIDGE C18 (150*19) mm, 5u;

[0813] Mobile phase: (A) 0.1% formic acid (B) acetonitrile;

[0814] Flow rate: 19 mL / min,

[0815] Gradient - (T / %B): 0 / 0, 2 / 0, 8 / 20, 10.9 / 20, 10.95 / 99, 13 / 99, 13.10 / 0, 16 / 0;

[0816] Solubility: ACN + H 2 O + THF.

[0817] Example 35

[0818] 5 - [[4 - [(4 - Amino - 4 - imino - butanoyl)amino] - 3 - fluoro - phenyl]sulfonylamino]thiazole - 4 - carboxylic acid

[0819]

[0820] a. 5 - [[4 - (3 - Chloropropionylamino) - 3 - fluoro - phenyl]sulfonyl - [(4 - methoxyphenyl)methyl]amino]thiazole - 4 - carboxylic acid

[0821]

[0822] At 0 °C, a solution of acetic anhydride (5 mL) in DCM (5 mL) was added to a stirred solution of 3 - chloropropionyl chloride (1.5 g, 3.04 mmol) in DCM (5 mL). After 10 minutes, a solution of 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 room temperature for 2 hours, quenched with ice water (20 mL) and extracted with DCM (2 × 10 mL). The organic layer was dried over Na 2 SO 4 dried, filtered and concentrated. The crude product was purified by flash chromatography (eluting with a solution of 60% ethyl acetate in petroleum ether) to give an off - white solid (600 mg, 33%).

[0823] M / z 584.40 (M + H) +

[0824] b. tert - Butyl 5 - [[4 - (3 - cyanopropionylamino) - 3 - fluoro - phenyl]sulfonyl - [(4 - methoxyphenyl)methyl]amino]thiazole - 4 - carboxylate

[0825]

[0826] At room temperature, sodium cyanide (76 mg, 1.54 mmol) was added to a stirred solution of 5-[[4-(3-chloropropionylamino)-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylic acid (600 mg, 1.02 mmol) in DMF (5 mL). The resulting reaction mixture was stirred at room temperature for 6 h and quenched with ice water (10 mL). The precipitate obtained was filtered, washed with Et 2 O (2 × 10 mL) and dried under high vacuum to give a brown solid (500 mg, 84%).

[0827] M / z 597.24 (M+Na) +

[0828] c. tert-Butyl 5-[[4-[(4-amino-4-imino-butanoyl)amino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

[0829]

[0830] HCl gas was bubbled through a stirred solution of tert-butyl 5-[[4-(3-cyanopropionylamino)-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate (400 mg, 0.69 mmol) in ethanol:Et 2 O (1:4, 10 mL) for 2 h. The resulting reaction mixture was kept at 4 °C for 16 h. The volatile components were then evaporated under reduced pressure. The residue was dissolved in ethanol (5 mL) and NH 3 gas was bubbled through it for 20 min. The volatile components were evaporated under reduced pressure. The crude product obtained was triturated with diethyl ether (2 × 5 mL) and dried under high vacuum to give a brown solid, which was used in the next step without further purification (350 mg, crude).

[0831] M / z 592.43 (M+H) +

[0832] d. 5-[[4-[(4-amino-4-imino-butanoyl)amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylic acid

[0833]

[0834] At 0 °C, TFA:H 2O(95:5, 3 mL) was added to tert-butyl 5-[[4-[(4-amino-4-imino-butanoyl)amino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate (350 mg, 0.592 mmol). The resulting reaction mixture was stirred at room temperature for 6 h and concentrated under reduced pressure. The obtained crude product was triturated with diethyl ether (2 × 5 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%).

[0835] 1 H 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).

[0836] M / z 415.93 (M + H) +

[0837] LC-MS conditions:

[0838] Column: Acquity BEH C18 (50 mm x 2.1 mm, 1.7 um);

[0839] Mobile phase: A: aqueous solution of 0.05% formic acid; B: ACN solution of 0.05% formic acid;

[0840] Time (min) / % B: 0 / 3, 0.4 / 3, 2 / 98, 3.4 / 98, 3.5 / 3, 4 / 3;

[0841] Column temperature: 35 °C;

[0842] Flow rate: 0.6 mL / min.

[0843] Prep.HPLC conditions:

[0844] Column: SymmetryC18 (300 * 19) mm, 7 u;

[0845] Mobile phase: (A) 0.1% formic acid (B) acetonitrile;

[0846] Flow rate: 19 mL / min;

[0847] Gradient (T / % B): 0 / 5, 1 / 5, 7 / 20, 10.1 / 20, 10.1 / 99, 13 / 99, 13.1 / 5, 16 / 5;

[0848] Solubility: ACN + H 2 O + THF + DMSO + concentrated FA.

[0849] Example 36

[0850] 5-[[4-[3-(4,5-Dihydro-1H-imidazol-2-yl)propanoyl amino]-3-fluoro-phenyl]sulfonyl amino]thiazole-4-carboxylic acid

[0851]

[0852] a. 5-[[4-[3-(4,5-Dihydro-1H-imidazol-2-yl)propanoyl amino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylic acid

[0853]

[0854] At 0 °C, HCl gas was bubbled into a stirred solution of tert-butyl 5-[[4-(3-cyanopropanoyl amino)-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate (310 mg, 0.53 mmol) in ethanol:Et 2 O (1:4, 15 mL) for 2 hours. The resulting reaction mixture was kept in the refrigerator for 16 hours. Then the volatile components were evaporated under reduced pressure. The resulting residue was dissolved in ethanol (5 mL). Then ethylenediamine (32 mg, 0.53 mmol) was added at room temperature. The resulting reaction mixture was stirred at room temperature for 8 hours and concentrated under reduced pressure. The resulting crude product was triturated with n-pentane (2 × 5 mL) and dried under high vacuum to give a brown solid, which was used in the next step without further purification (400 mg, crude).

[0855] M / z 618.46 (M + H) +

[0856] b. 5-[[4-[3-(4,5-Dihydro-1H-imidazol-2-yl)propanoyl amino]-3-fluoro-phenyl]sulfonyl amino]thiazole-4-carboxylic acid

[0857]

[0858] At 0 °C, TFA:H 2O(95:5, 3 mL) was added to 5-[[4-[3-(4,5-dihydro-1H-imidazol-2-yl)propanoyl]amino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylic acid (280 mg, 0.45 mmol). The resulting reaction mixture was stirred at room temperature for 4 hours and concentrated under reduced pressure. The obtained crude product was triturated with diethyl ether (2 × 5 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%).

[0859] 1 H NMR (300 MHz, DMSO-d 6 ) 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).

[0860] M / z 441.98 (M + H) +

[0861] LC-MS conditions:

[0862] Column: Acquity BEH C18 (50 mm x 2.1 mm, 1.7 um);

[0863] Mobile phase: A: Aqueous solution of 0.05% formic acid; B: ACN solution of 0.05% formic acid;

[0864] Time (min) / % B: 0 / 3, 0.4 / 3, 3.2 / 98, 3.8 / 98, 4.2 / 3, 4.5 / 3;

[0865] Column temperature: 35 °C;

[0866] Flow rate: 0.6 mL / min.

[0867] Prep.HPLC conditions:

[0868] Column: Symmetry C18 (300 * 19) mm, 7 u;

[0869] Mobile phase: (A) 0.1% formic acid (B) acetonitrile;

[0870] Flow rate: 19 mL / min;

[0871] Gradient (T / % B): 0 / 5, 1 / 5, 7 / 30, 8.7 / 30, 8.75 / 99, 11 / 99, 11.1 / 5, 13 / 5;

[0872] Solubility: ACN + H 2 O + THF。

[0873] Example 37

[0874] 5-[[4-[(3-Amino-3-imino-2-methyl-propionyl)amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylic acid

[0875]

[0876] a. tert-Butyl 5-[[4-(2-chloropropionylamino)-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

[0877]

[0878] At 0 °C, 2-chloropropionyl 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-carboxylate (3 g, 6.0 mmol) in DCM (50 mL). The resulting reaction mixture was stirred at room temperature for 2 h and concentrated under reduced pressure. The residue was triturated with diethyl ether (2 × 50 mL) and pentane (2 × 50 mL) and dried under reduced pressure to give an off-white solid (3.4 g, 95%).

[0879] M / z 606.28 (M+Na) + ; 582.75 (M-H) -

[0880] b. tert-Butyl 5-[[4-(2-cyanopropionylamino)-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

[0881]

[0882] At room temperature, NaCN (570 mg, 11.6 mmol) was added to a solution of tert-butyl 5-[[4-(2-chloropropionylamino)-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate (3.4 g, 5.8 mmol) in DMF (35 mL). The resulting reaction mixture was stirred at room temperature for 16 h and quenched with ice water. The precipitate obtained was filtered and dried under high vacuum. The crude product was purified by silica gel chromatography (eluting with a petroleum ether solution of 40% EtOAc) to give an off-white solid (1.5 g, 44%).

[0883] M / z 597.29 (M+Na)+ ; 573.62 (M-H) -

[0884] c. tert-Butyl 5-[[3-fluoro-4-[[3-(hydroxyamino)-3-imino-2-methyl-propanoyl]amino]phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

[0885]

[0886] At room temperature, 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). The resulting reaction mixture was stirred at 65 °C for 1 h, cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure to give a pale yellow gummy material, which was used for the next step without further purification (1.5 g, crude).

[0887] M / z 608.48 (M+H) +

[0888] d. tert-Butyl 5-[[4-[(3-amino-3-imino-2-methyl-propanoyl)amino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

[0889]

[0890] Iron powder (193 mg, 3.4 mmol) was added to a solution of tert-butyl 5-[[3-fluoro-4-[[3-(hydroxyamino)-3-imino-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 the mixture was heated to reflux for 30 min. Then a solution of 1N HCl (0.3 mL) in ethanol:water (1:1, 3 mL) was added to the reaction mixture over 30 min. The reaction mixture was stirred at 70 °C for an additional 1 h, cooled to room temperature and filtered through celite. The celite pad was washed with ethanol (2 × 10 mL). The filtrate was concentrated under reduced pressure to give a pale yellow liquid, which was used for the next step without further purification (340 mg, crude).

[0891] M / z 592.28 (M+H)+

[0892] e. 5-[[4-[(3-Amino-3-imino-2-methyl-propionyl)amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylic acid

[0893]

[0894] At room temperature, tert-butyl 5-[[4-[(3-amino-3-imino-2-methyl-propionyl)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). The reaction mixture was stirred at room temperature for 3 h and concentrated under reduced pressure (30 °C below the bath temperature). The residue was triturated with diethyl ether (2 × 5 mL) and dried under high vacuum. The crude product was purified by preparative HPLC to give the title product (48.2 mg) as an off-white solid.

[0895] 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).

[0896] M / z 416.34 (M+H) +

[0897] LC-MS conditions:

[0898] Column: Acquity BEH C18 (50 mm x 2.1 mm, 1.7 um);

[0899] Mobile phase: A: aqueous solution of 0.05% formic acid; B: ACN solution of 0.05% formic acid;

[0900] Time (min) / % B: 0 / 3, 0.4 / 3, 3.2 / 98, 3.8 / 98, 4.2 / 3, 4.5 / 3;

[0901] Column temperature: 35 °C;

[0902] Flow rate: 0.6 mL / min.

[0903] Prep.HPLC conditions:

[0904] Column used: Symmetry C18 (300×19) mm, 7u;

[0905] Mobile phase: (A) 0.1% formic acid (B) acetonitrile;

[0906] Flow rate: 19 mL / min;

[0907] Gradient - (T / %B): 0 / 5, 1 / 5, 8 / 50, 8.1 / 99, 11 / 99, 11.1 / 5, 14 / 5;

[0908] Solubility: ACN + H 2 O + concentrated FA.

[0909] Example 38

[0910] 5 - [[3 - fluoro - 4 - [[2 - (2 - iminoimidazolidin - 1 - yl)acetyl]amino]phenyl]sulfonylamino]thiazole - 4 - carboxylic acid

[0911]

[0912] a. tert - butyl 5 - [[3 - fluoro - 4 - [[2 - (2 - thioimidazolidin - 1 - yl)acetyl]amino]phenyl]sulfonyl - [(4 - methoxyphenyl)methyl]amino]thiazole - 4 - carboxylate

[0913]

[0914] At 75 °C, 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 30 minutes. The resulting reaction mixture was stirred at 75 °C for 2.5 hours. Then bis(imidazol - 1 - yl)methanethione (1.56 g,

[0915] 8.77 mmol) was added at 75 °C and the reaction was stirred at the same temperature for 1 hour, then concentrated under reduced pressure. The resulting crude compound was diluted with EtOAc (50 mL) and washed with water (10 mL) and brine solution (10 mL). The organic layer was dried over Na 2 SO 4 dried, filtered and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with a 60% EtOAc in petroleum ether solution) to give a light brown solid (200 mg, 36%).

[0916] M / z 636.20 (M + H) +

[0917] b. tert-Butyl 5-[[3-fluoro-4-[[2-(2-iminoimidazolidin-1-yl)acetyl]amino]phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

[0918]

[0919] At -30 °C under a nitrogen atmosphere, Ag(OTf) (121 mg, 0.47 mmol) and a saturated solution of NH 3 in THF (5 mL) were added to a stirred solution of tert-butyl 5-[[3-fluoro-4-[[2-(2-thioimidazolidin-1-yl)acetyl]amino]phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate (200 mg, 0.31 mmol) in CH 2 Cl 2 (10 mL). The resulting reaction mixture was stirred at room temperature for 16 h, quenched with MeOH (2 mL) and stirred at room temperature for 10 min. The reaction mixture was filtered through a pad of Celite, and the pad was washed with CH 2 Cl 2 (2 × 10 mL). The filtrate was concentrated under reduced pressure to give a dark brown liquid which was used without further purification in the next step (300 mg, crude).

[0920] M / z 619.48 (M+H) +

[0921] c. 5-[[3-fluoro-4-[[2-(2-iminoimidazolidin-1-yl)acetyl]amino]phenyl]sulfonylamino]thiazole-4-carboxylic acid

[0922]

[0923] At 0 °C, TFA:H 2 O (9:1, 5 mL) was added to tert-butyl 5-[[3-fluoro-4-[[2-(2-iminoimidazolidin-1-yl)acetyl]amino]phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate (270 mg, 0.43 mmol). The resulting reaction mixture was stirred at room temperature for 3 h and concentrated under reduced pressure. The resulting crude product was triturated with diethyl ether (2 × 5 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).

[0924] 1 H NMR (400 MHz, DMSO-d 6)δ13.32(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=8.0Hz,J=7.6Hz),7.38(1H,s),4.11(2H,s),3.74 - 3.67(2H,m),3.61 - 3.55(2H,m).

[0925] M / z 443.24(M + H) +

[0926] LC-MS conditions:

[0927] Column: Acquity BEH C18(50mm x 2.1mm, 1.7um);

[0928] Mobile phase: A: Aqueous solution of 0.05% formic acid; B: ACN solution of 0.05% formic acid;

[0929] Time (min) / % B: 0 / 3, 0.4 / 3, 3.2 / 98, 3.8 / 98, 4.2 / 3, 4.5 / 3;

[0930] Column temperature: 35 °C,

[0931] Flow rate: 0.6 mL / min

[0932] Prep.HPLC conditions:

[0933] Column used: Atlantis T3(250 * 19)mm, 5u;

[0934] Mobile phase: (A) 0.1% formic acid (B) acetonitrile;

[0935] Flow rate: 19 mL / min;

[0936] Gradient - (T / % B): 0 / 5, 1 / 5, 7 / 30, 8.25 / 30, 8.3 / 99, 11 / 99, 11.1 / 5, 14 / 5;

[0937] Solubility: ACN + H 2 O + THF

[0938] Example 39

[0939] 5 - [[4 - [[2 - [[N-(2 - aminoethyl)formamidinyl]amino]acetyl]amino]-3 - fluoro - phenyl]sulfonylamino]thiazole - 4 - carboxylic acid

[0940]

[0941] a. tert-Butyl 5-[[4-[[2-[2-(tert-Butoxycarbonylamino)ethylthiocarbamoyl]amino]acetyl]amino]-3-fluorophenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

[0942]

[0943] At room temperature, bis(imidazol-1-yl)methanethione (97 mg, 0.54 mmol) was added to a stirred solution of tert-butyl 5-[[4-[(2-aminoacetyl)amino]-3-fluorophenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate (200 mg, 0.36 mmol) in DCM (10 mL). The reaction mixture was stirred at room temperature 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 a petroleum ether solution of 60% EtOAc) to give a brown solid (80 mg, 29%).

[0944] M / z 753.43 (M+H) +

[0945] b. tert-Butyl 5-[[4-[[2-[[N-[2-(tert-Butoxycarbonylamino)ethyl]formamidinyl]amino]acetyl]amino]-3-fluorophenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

[0946]

[0947] At room temperature, Ag(OTf) (107 mg, 0.47 mmol) was added to a stirred solution of tert-butyl 5-[[4-[[2-[2-(tert-Butoxycarbonylamino)ethylthiocarbamoyl]amino]acetyl]amino]-3-fluorophenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate (220 mg, 0.27 mmol) in CH 2 Cl 2 (10 mL). The reaction mixture was stirred at room temperature for 15 min and a saturated solution of NH 3 in THF (5 mL) was added at -30 °C under a nitrogen atmosphere. The resulting reaction mixture was stirred at room temperature for 6 h, filtered through a pad of Celite, and washed with CH 2 Cl 2Wash the pad with (10 mL). Concentrate the filtrate and triturate the obtained crude material with n-pentane (10 mL) to give a brown solid, which is used for the next step without further purification.

[0948] M / z 736.40 (M+H) +

[0949] c. 5-[[4-[[2-[[N-(2-Aminoethyl)formamidinyl]amino]acetyl]amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylic acid

[0950]

[0951] At 0 °C, add TFA:H 2 O (9:1, 2 mL) to tert-butyl 5-[[4-[[2-[[N-[2-(tert-butoxycarbonylamino)ethyl]formamidinyl]amino]acetyl]amino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate (210 mg, 0.28 mmol). Stir the reaction mixture at room temperature for 4 h and concentrate it under reduced pressure. Triturate the obtained crude product with diethyl ether (2 × 5 mL) and dry it under high vacuum. Purify the crude product by preparative HPLC to give the title product (25 mg) as a brown solid.

[0952] 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).

[0953] M / z 460.30 (M+H) +

[0954] LC-MS conditions:

[0955] Column: Acquity BEH C18 (50 mm x 2.1 mm, 1.7 um);

[0956] Mobile phase: A: Aqueous solution of 0.05% formic acid; B: ACN solution of 0.05% formic acid;

[0957] Time (min) / % B: 0 / 3, 0.4 / 3, 3.2 / 98, 3.8 / 98, 4.2 / 3, 4.5 / 3;

[0958] Column temperature: 35 °C;

[0959] Flow rate: 0.6 mL / min.

[0960] Prep.HPLC conditions:

[0961] Column: Atlantis T3 (250*19) mm, 5u;

[0962] Mobile phase: (A) 0.1% formic acid (B) acetonitrile;

[0963] Flow rate: 19 mL / min;

[0964] Gradient (T / %B): 0 / 5, 1 / 5, 7 / 25, 12 / 30, 12.1 / 99, 15 / 99, 15.1 / 5, 18 / 5;

[0965] Solubility: ACN + H 2 O + THF + FA.

[0966] Compounds prepared by a method similar to that of Example 39 above using methylamine in step a and purified in a similar manner by preparative HPLC are shown in the following table:

[0967]

[0968] Example 41

[0969] 5-[[4-[[2-(2-Methylformohydrazide)acetyl]amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylic acid

[0970]

[0971] a. tert-Butyl 5-[[4-[[2-(2-tert-butoxycarbonylhydrazino)acetyl]amino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

[0972]

[0973] At room temperature, KI (1.17 g, 7.01 mmol) was added to a solution of tert-butyl 5-[[4-[(2-chloroacetyl)amino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate (2 g, 3.50 mmol) in DMF (20 mL). After 10 minutes, tert-butyl N-aminocarbamate (695 mg, 5.26 mmol) was added to the reaction mixture at the same temperature. The resulting reaction mixture was stirred at room temperature 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, washed with diethyl ether and dried under high vacuum to give a pale yellow solid, which was used in the next step without further purification (1.2 g, 52%).

[0974] M / z 666.48(M+H) +

[0975] b. 5-[[3-Fluoro-4-[(2-hydrazinoacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylic acid

[0976]

[0977] At room temperature, TFA (4 mL) was added to tert-butyl 5-[[4-[[2-(2-tert-butoxycarbonylhydrazino)acetyl]amino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate (1.2 g, 1.80 mmol). The reaction mixture was stirred at room temperature for 16 h and concentrated under reduced pressure. The resulting crude product was triturated with diethyl ether (3 × 10 mL) to give a pale yellow solid, which was used in the next step without further purification (1 g, crude).

[0978] M / z 390.32(M+H) +

[0979] c. 5-[[4-[[2-(2-formamidohydrazino)acetyl]amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylic acid

[0980]

[0981] At room temperature, 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). The resulting reaction mixture was stirred at room temperature for 16 h, concentrated under reduced pressure and water (5 mL) was added to the residue. The precipitate obtained was filtered and washed with Et 2 O (2 × 10 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).

[0982] 1 1H 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 = 4.4 Hz).

[0983] M / z 432.37 (M + H) +

[0984] LC-MS conditions:

[0985] Column: Acquity BEH C18 (50 mm x 2.1 mm, 1.7 um);

[0986] Mobile phase: A: aqueous solution of 0.05% formic acid; B: ACN solution of 0.05% formic acid;

[0987] Time (min) / % B: 0 / 3, 0.4 / 3, 3.2 / 98, 3.8 / 98, 4.2 / 3, 4.5 / 3;

[0988] Column temperature: 35 °C;

[0989] Flow rate: 0.6 mL / min.

[0990] Prep.HPLC conditions:

[0991] Column: X BRIDGE C18 (150 * 19) mm, 5u;

[0992] Mobile phase (A) 0.1% formic acid (B) acetonitrile;

[0993] Flow rate: 19 mL / min;

[0994] Gradient (T / %B): (T / %B): 0 / 0, 3 / 0, 8.8 / 33, 9 / 33, 9.10 / 99, 12 / 99, 12.10 / 0, 15 / 0;

[0995] Solubility: ACN + H 2 O + THF + DMSO + FA.

[0996] Example 42

[0997] 5-[[3-Fluoro-4-[(2-guanidinooxyacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylic acid

[0998]

[0999] a. tert-Butyl 5-[[4-[[2-(tert-butoxycarbonylamino)oxyacetyl]amino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

[1000]

[1001] Under 0 °C and an argon atmosphere, 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), and the mixture was stirred at room temperature for 30 minutes. Then, under 0 °C and an argon atmosphere, 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 THF (10 mL) was added to the above reaction mixture. The resulting reaction mixture was stirred at room temperature for 1.5 hours, quenched with ice water (20 mL) and extracted with ethyl acetate (2 × 20 mL). The combined organic layers were dried over Na 2 SO 4 dried, filtered and concentrated under reduced pressure. The crude product was purified by column chromatography (eluted with a petroleum ether solution of 30% ethyl acetate) to obtain a yellow solid (350 mg, 59%).

[1002] M / z 667.10 (M + H) +

[1003] b. 5-[[4-[(2-Aminooxyacetyl)amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylic acid

[1004]

[1005] At 0 °C, TFA:H 2O(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). The reaction mixture was stirred at room temperature for 4 h and concentrated under reduced pressure. The resulting crude product was triturated with diethyl ether (3 × 10 mL) to give an off-white solid, which was used for the next step without further purification (200 mg, crude).

[1006] M / z 390.95 (M+H) +

[1007] c. 5-[[3-Fluoro-4-[(2-guanidinooxyacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylic acid

[1008]

[1009] At 0 °C, 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 in DMF (6 mL). The resulting reaction mixture was stirred at room temperature for 6 h, concentrated under reduced pressure and water (5 mL) was added to the residue. The resulting precipitate was filtered, washed with Et 2 O (2 × 10 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%).

[1010] 1 H NMR (300 MHz, DMSO-d 6 ) δ 13.50 (1H, brs), δ 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).

[1011] M / z 433.30 (M+H) +

[1012] LC-MS conditions:

[1013] Column: Acquity BEH C18 (50 mm x 2.1 mm, 1.7 um);

[1014] Mobile phase: A: Aqueous solution of 0.05% formic acid: Acetonitrile solution of 0.05% formic acid;

[1015] Time (min) / % B: 0 / 3, 0.4 / 3, 3.2 / 98, 3.8 / 98, 4.2 / 3, 4.5 / 3;

[1016] Column temperature: 35 °C;

[1017] Flow rate: 0.6 mL / min.

[1018] Prep.HPLC conditions:

[1019] Column: X BRIDGE C18 (150*19) mm, 5u;

[1020] Mobile phase (A) 0.1% formic acid (B) acetonitrile;

[1021] Flow rate: 19 mL / min;

[1022] Gradient (T / % B): - (T / % B): 0 / 0, 3 / 0, 8.8 / 33, 9 / 33, 9.10 / 99, 12 / 99, 12.10 / 0, 15 / 0;

[1023] Solubility: ACN + H 2 O + THF + DMSO + FA.

[1024] Example 43

[1025] 5 - [[4 - [[(2E)-2-(Formamidinohydrazono)acetyl]amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylic acid

[1026]

[1027] a. tert-Butyl 5-[[4-[[4-[4-[(4-tert-Butoxycarbonylthiazol-5-yl)-[(4-methoxyphenyl)methyl]sulfamoyl]-2-fluoro-phenylamino-4-oxo-but-2-enoyl]amino]-3-fluoro-phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

[1028]

[1029] At 0 °C, a solution of but-2-enedioyl dichloride (0.18 g, 1.2 mmol) in DCM (20 mL) was added to a solution of tert-butyl 5-[(4-amino-3-fluorophenyl)sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate (1 g, 2.0 mmol) in DCM (10 mL). The resulting reaction mixture was stirred at room temperature for 6 h, diluted with DCM, and washed with water (2 × 10 mL) and brine (2 × 10 mL) solutions. The combined organic layers were dried over Na 2 SO 4 dried, filtered, and concentrated under reduced pressure. The crude material was purified by flash chromatography (eluting with a petroleum ether solution of 50% ethyl acetate) to give a pale yellow solid (500 mg, 23%).

[1030] M / z 1067.05 (M+H) +

[1031] b. tert-Butyl 5-[[3-fluoro-4-(oxoacetylamino)phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

[1032]

[1033] At 78 °C, a solution of tert-butyl 5-[[4-[[4-[4-[(4-tert-butoxycarbonylthiazol-5-yl)-[(4-methoxyphenyl)methyl]sulfamoyl]-2-fluoro-phenylamino-4-oxo-but-2-enoyl]amino]-3-fluorophenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate (500 mg, 0.46 mmol) in DCM / MeOH (3:1, 40 mL) was purged with ozone gas for 1 h. Then, at the same temperature, DMS (2 mL) was added to the reaction mixture, and the resulting mixture was stirred at room temperature for 2 h. The crude reaction mixture was used in the next step without further purification.

[1034] c. tert-Butyl 5-[[4-[[(2E)-2-(formamidinohydrazono)acetyl]amino]-3-fluorophenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

[1035]

[1036] At room temperature, 1-aminoguanidine hydrochloride (30 mg, 0.27 mmol) was added to a stirred solution of tert-butyl 5-[[3-fluoro-4-(oxoacetylamino)phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate (100 mg, 0.18 mmol) in DCM / MeOH (1:1, 10 mL). The resulting reaction mixture was stirred at room temperature for 16 h and concentrated under reduced pressure. The crude product was used in the next step without further purification (100 mg, crude).

[1037] M / z 522.1(M+H) +

[1038] d. 5-[[4-[[(2E)-2-(formamidinohydrazono)acetyl]amino]-3-fluorophenyl]sulfonylamino]thiazole-4-carboxylic acid

[1039]

[1040] At 0 °C, TFA / H 2 O (9:1, 1.0 mL) was added to tert-butyl 5-[[4-[[(2E)-2-(formamidinohydrazono)acetyl]amino]-3-fluorophenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate (80 mg, 0.13 mmol), stirred at room temperature for 4 h, and concentrated under reduced pressure. The resulting crude product was triturated with diethyl ether (2 × 5 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).

[1041] 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.68 (1H, brs), 8.05 (1H, s), 7.80 (1H, dd, J = 8.4 Hz, J = 8.0 Hz), 7.53 - 7.47 (2H, m), 7.19 (1H, s), 6.80 - 6.00 (4H, brs).

[1042] M / z 430.31(M+H) +

[1043] LC-MS conditions:

[1044] Column: Acquity BEH C18 (50 mm x 2.1 mm, 1.7 um);

[1045] Mobile phase: A: aqueous solution of 0.05% formic acid; B: ACN solution of 0.05% formic acid;

[1046] Time (min) / % B: 0 / 3, 0.4 / 3, 3.2 / 98, 3.8 / 98, 4.2 / 3, 4.5 / 3;

[1047] Column temperature: 35 °C;

[1048] Flow rate: 0.6 mL / min.

[1049] Prep.HPLC conditions:

[1050] Column: Atlantis T3 (250*19) mm, 5u;

[1051] Mobile phase: (A) 0.1% formic acid (B) acetonitrile;

[1052] Flow rate: 19 mL / min;

[1053] Gradient (T / % B): 0 / 5, 1 / 5, 7 / 25, 12 / 30, 12.1 / 99, 15 / 99, 15.1 / 5, 18 / 5;

[1054] Solubility: ACN + H 2 O + THF + FA

[1055] Example 44

[1056] 5-[(4-Guanidinophenyl)sulfonylamino]thiazole-4-carboxylic acid

[1057]

[1058] a. tert-Butyl 5-[[4-[[N,N'-bis(tert-butoxycarbonyl)formamidinyl]amino]phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

[1059]

[1060] Under 0 °C and argon atmosphere, 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). After 30 minutes, under 0 °C and argon atmosphere, 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. The resulting reaction mixture was stirred at room temperature for 16 hours, quenched with ice water (20 mL) and extracted with ethyl acetate (2 × 20 mL). The combined organic layers were washed with Na 2 SO 4Dry, filter and concentrate under reduced pressure. Purify the crude by trituration with diethyl ether (2 x 5 mL) to give a pale yellow solid which is used for the next step without further purification (150 mg, crude).

[1061] M / z 662.03 (M+H-Boc) +

[1062] b. 5-[(4-Guanidinophenyl)sulfonylamino]thiazole-4-carboxylic acid

[1063]

[1064] At room temperature, add TFA:H 2 O (9:1, 2 mL) to tert-butyl 5-[[4-[[N,N'-bis(tert-butoxycarbonyl)formamidinyl]amino]phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate (150 mg, 0.22 mmol). Stir the resulting mixture for 4 h and concentrate under reduced pressure. Triturate the obtained crude product with diethyl ether (2 x 5 mL) and dry under high vacuum. Purify the crude product by preparative HPLC to give the title product as an off-white solid (22 mg, 28%).

[1065] 1 H NMR (400 MHz, DMSO-d 6 ) 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).

[1066] M / z 342.29 (M+H) +

[1067] LC-MS conditions:

[1068] Column: Acquity BEH C18 (50 mm x 2.1 mm, 1.7 um);

[1069] Mobile phase: A: aqueous solution of 0.05% formic acid; B: ACN solution of 0.05% formic acid;

[1070] Time (min) / % B: 0 / 3, 0.4 / 3, 3.2 / 98, 3.8 / 98, 4.2 / 3, 4.5 / 3;

[1071] Column temperature: 35 °C,

[1072] Flow rate: 0.6 mL / min.

[1073] Prep.HPLC conditions:

[1074] Column: Symmetry C18 (300*19) mm, 7u;

[1075] Mobile phase: (A) 0.1% formic acid (B) acetonitrile;

[1076] Flow rate: 19 mL / min;

[1077] Gradient (T / %B): 0 / 2, 1 / 1, 8 / 30, 9.10 / 99, 12 / 99, 12.10 / 2, 15 / 2;

[1078] Solubility: ACN+H 2 O+DMSO.

[1079] Example 45

[1080] 5-[[3-Fluoro-4-[[(2-guanidinoacetyl)amino]methyl]phenyl]sulfonylamino]thiazole-4-carboxylic acid

[1081]

[1082] a. tert-Butyl 5-[(3-fluoro-4-ethenyl-phenyl)sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

[1083]

[1084] Purge 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) with argon for 15 minutes. Then add 4,4,5,5-tetramethyl-2-ethenyl-1,3,2-dioxaborolane (0.99 g, 6.45 mmol), K 2 CO 3 (1.11 g, 8.07 mmol), PdCl 2 (PPh 3 ) 2 (0.37 g, 0.53 mmol) under an argon atmosphere. Heat the resulting reaction mixture in a sealed vial to 85 °C and maintain for 24 hours. Cool the reaction mixture to room temperature and filter through a pad of diatomaceous earth (washed with EtOAc (2×50 mL)). Concentrate the organic layer under reduced pressure. Dissolve the resulting crude compound in ethyl acetate (50 mL) and wash with water (50 mL) and brine solution (50 mL). Wash the organic layer with Na 2 SO 4Dry, filter and concentrate under vacuum. Purify the crude compound by flash chromatography (eluting with a petroleum ether solution of 20% ethyl acetate) to give an off-white solid (1.5 g, 55%).

[1085] M / z 505.1 (M+H) +

[1086] b. tert-Butyl 5-[(3-fluoro-4-formyl-phenyl)sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

[1087]

[1088] Add NaIO 4 (8.51 g, 39.8 mmol) and OsO 4 (1.68 g, 6.63 mmol) to a solution of tert-butyl 5-[(3-fluoro-4-vinyl-phenyl)sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate (6.7 g, 13.2 mmol) in CH3CN:H 2 2O:CCl4 4 (1:1:1, 60 mL). Stir the resulting reaction mixture at room temperature for 4 h. Add water (30 mL) and extract the mixture with ethyl acetate (2×100 mL). Dry the combined organic extracts over Na 2 2SO 4 , filter and concentrate. Purify the crude material by flash chromatography (eluting with a petroleum ether solution of 22% ethyl acetate) to give an off-white solid (4.5 g, 66%).

[1089] M / z 507.4 (M+H) +

[1090] c. tert-Butyl 5-[[3-fluoro-4-[hydroxyiminomethyl]phenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

[1091]

[1092] Add a solution of tert-butyl 5-[(3-fluoro-4-formyl-phenyl)sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate (3.6 g, 7.10 mmol) in EtOH (20 mL) to a stirred solution of hydroxylamine hydrochloride (593.8 mg, 8.52 mmol) and ammonium chloride (454.9 mg, 8.52 mmol) in H 2In an EtOH (4:1, 30 mL) solution. The resulting reaction mixture was stirred at room temperature for 4 h. Water (20 mL) was added and the mixture was extracted with ethyl acetate (2 × 50 mL). The combined organic extracts were dried over Na 2 SO 4 , filtered and concentrated. The crude material was purified by flash chromatography (eluting with a petroleum ether solution of 40% ethyl acetate) to afford an off-white solid (2.8 g, 75%).

[1093] M / z 522.1 (M+H) +

[1094] d. tert-Butyl 5-[[4-(aminomethyl)-3-fluorophenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

[1095]

[1096] At room temperature, zinc powder (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). The resulting reaction mixture was stirred at room temperature for 16 h. Ice water was added and the mixture was extracted with ethyl acetate (2 × 50 mL). The combined organic extracts were dried over Na 2 SO 4 , filtered and concentrated. The crude material was purified by trituration with diethyl ether (2 × 10 mL) to afford a yellow solid (2 g, 73%).

[1097] M / z 508.1 (M+H) +

[1098] e. tert-Butyl 5-[[4-[[[2-(tert-butoxycarbonylamino)acetyl]amino]methyl]-3-fluorophenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate

[1099]

[1100] Under an argon atmosphere, 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-fluorophenyl]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). The resulting reaction mixture was stirred at room temperature for 4 h. Ice water (10 mL) was added and the mixture was extracted with ethyl acetate (2 × 20 mL). The combined organic extracts were dried over Na 2 SO 4 , filtered and concentrated. The crude material was purified by flash chromatography (eluting with a petroleum ether solution of 50% ethyl acetate) to give an off-white solid (500 mg, 63%).

[1101] M / z 508.1 (M+H) +

[1102] f. 5-[[4-[[(2-Aminoacetyl)amino]methyl]-3-fluorophenyl]sulfonylamino]thiazole-4-carboxylic acid trifluoroacetate

[1103]

[1104] At room temperature, TFA (5 mL) was added to tert-butyl 5-[[4-[[[2-(tert-butoxycarbonylamino)acetyl]amino]methyl]-3-fluorophenyl]sulfonyl-[(4-methoxyphenyl)methyl]amino]thiazole-4-carboxylate (500 mg, 0.75 mmol) and the mixture was stirred for 4 h. The TFA was evaporated under reduced pressure. The resulting crude product was triturated with diethyl ether (2 × 10 mL) and dried under high vacuum to give an off-white solid (250 mg, 85%).

[1105] M / z 389.1 (M+H) +

[1106] g. 5-[[3-Fluoro-4-[[(2-guanidinoacetyl)amino]methyl]phenyl]sulfonylamino]thiazole-4-carboxylic acid

[1107]

[1108] Pyrazole-1-carboximidamide 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-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylic acid trifluoroacetate (250 mg, 0.64 mmol) in DMF (6 mL). The resulting reaction mixture was stirred at room temperature for 18 h and concentrated under reduced pressure. Water (5 mL) was added to the residue. The resulting precipitate was filtered and washed with diethyl ether (2 × 5 mL). The crude product was purified by preparative HPLC to give the title compound as a white solid (70 mg, 25%).

[1109] 1 H NMR (500 MHz, DMSO-d 6 ) δ 13.43 (1H, brs), 8.60 (1H, brs), 8.08 (1H, s), 7.51 - 7.42 (3H, m), 7.50–7.10 (4H, brs), 4.33 (2H, s), 3.85 (2H, s).

[1110] M / z 431.0 (M + H) +

[1111] LC-MS conditions:

[1112] Column: Acquity UPLC BEH C18 (50 mm x 2.1 mm, 1.7 um);

[1113] Mobile phase: A: aqueous solution of 0.1% formic acid; B: acetonitrile solution of 0.1% formic acid;

[1114] Flow rate: 0.8 mL / min

[1115] Time (min) / % B: 0 / 2, 0.4 / 2, 2.2 / 98, 2.6 / 98, 2.61 / 2, 3.0 / 2.

[1116] Column temperature: 60 °C.

[1117] Prep.HPLC conditions:

[1118] Column: KROMASIL-C18 (150*25MM), 10u;

[1119] Mobile phase: 0.05% formic acid in H 2 O: acetonitrile solution;

[1120] Flow rate: 25 mL / min

[1121] Gradient (T / %B): 0 / 5, 1 / 5, 7 / 40, 7.1 / 98, 9 / 98, 9.1 / 5, 11 / 5;

[1122] Solubility: ACN + H 2 O + THF + DMSO

[1123] The compounds prepared using a method similar to that described in Example 45 above and purified in a similar manner by preparative HPLC are shown in the following table:

[1124]

[1125] Example 47: Activity of the Compounds of the Invention

[1126] Experiments were conducted to determine:

[1127] (1) The inhibitory activity of the compounds of the invention against the MBL enzyme;

[1128] (2) The plasma protein binding of the compounds of the invention; and

[1129] (3) The plasma stability of the compounds of the invention.

[1130] Details of the protocol for each set of experiments are as follows:

[1131] 1. Enzyme Inhibition

[1132] In Vitro Enzyme Inhibition Assay

[1133] In a 96-well microtiter plate, an enzyme inhibition assay was performed using purified MBL enzymes (NDM-1; VIM-1; VIM-2; IMP-1) in 10 mM HEPES buffer at pH 7.5. Imipenem (300 μM) was used as the substrate, and the hydrolysis of the substrate was monitored every 30 seconds at UV 299 nm for 10 minutes using a PerkinElmer Envision UV fluorescence microplate reader. The hydrolysis rate data for a series of inhibitors were analyzed using Dotmatics database software, and the calculated IC 50 values were converted to Ki values using the Cheng-Prusoff equation:

[1134] Ki = IC 50 / (1 + ([S] / K m )

[1135] 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. The compounds were diluted in DMSO.

[1136] The average Ki values from multiple experiments are presented as follows. The experimental results are given using the following ranges:

[1137] - For NDM-1, a Ki value < 0.05 μM is labeled as (A); a Ki value of 0.05 μM to 0.2 μM is labeled as (B); a Ki value > 0.2 μM (0.2 μM to 2 μM) is labeled as (C).

[1138] - For VIM-1, a Ki value < 0.2 μM is labeled as (A); a Ki value of 0.2 μM to 0.5 μM is labeled as (B); a Ki value > 0.5 μM (0.5 μM to 1 μM) is labeled as (C).

[1139] - For VIM-2, a Ki value < 0.02 μM is labeled as (A); a Ki value of 0.02 μM to 0.05 μM is labeled as (B); a Ki value > 0.05 μM (0.05 μM to 0.15 μM) is labeled as (C).

[1140] - For IMP-1, a Ki value < 0.5 μM is labeled as (A); a Ki value of 0.5 μM to 1 μM is labeled as (B); a Ki value > 1 μM (1 μM to 10 μM) is labeled as (C).

[1141] The Ki values of the compounds of the present invention

[1142]

[1143]

[1144]

[1145] 2. Antimicrobial susceptibility testing

[1146] The antibiotic activity of β-lactam antibiotics against MBL-expressing bacteria in the presence of the compounds of the present invention

[1147] The experiment was conducted using the "broth microdilution method" according to the protocol M07-A8 of the Clinical and Laboratory Standards Institute (CLSI). In cation-adjusted Mueller-Hinton broth (CAMHB), serial dilutions of the β-lactam antibiotic (meropenem) were prepared in a 96-well plate; the concentration range was limited to 0.03 mg / L to 512 mg / L. The compound was added at a constant concentration of 8 mg / L. The bacterial inoculum of each strain (clinical isolate) was adjusted to a McFarland turbidity standard of 0.5 in physiological serum (0.9% NaCl), then diluted 1:100 in CAMHB and added to each well, resulting in a final bacterial cell count of 5×10 5CFU per well. After incubation in the heating chamber at 37 °C for 18 to 20 hours, growth inhibition was evaluated based on the absence of any bacterial amplification.

[1148] The minimum inhibitory concentration (MIC) was taken as the lowest antibiotic concentration at which the test organism did not show visible growth; the results were confirmed by measuring the optical density (OD) at 600 nm in a spectrophotometer.

[1149] The compounds of the present invention were tested at a constant concentration of 8 μg / mL. The clinical strains used in these potentiation experiments were NTBC020 (Escherichia coli strain expressing NDM-1, TEM-1, and CTX-M-15), NTBC035-2 (Klebsiella pneumoniae strain expressing NDM-1, CMY-4, and SHV-11), NTBC104-1 (Klebsiella pneumoniae strain expressing NDM-1 and SHV-11), NTBC123 (Klebsiella pneumoniae strain expressing NDM-1), NTBC018 (Citrobacter freundii (C. freundii) strain expressing VIM-2), NTBC024 (Klebsiella pneumoniae strain expressing VIM-19, TEM-1, and CTX-M-3), NTBC042 (Escherichia coli strain expressing VIM-1, TEM-1, CTX-M-15, SHV-12), NTBC055 (Escherichia coli strain expressing VIM-1, NTBC062 (Klebsiella pneumoniae strain expressing IMP-1 and TEM-1), and NTBC039 (Klebsiella oxytoca strain expressing IMP-28).

[1150] The results are shown below. The data were segmented as follows: MIC values < 1 μg / mL were labeled as (A); MIC values of 1 μg / mL to 2 μg / mL were labeled as (B); MIC values > 2 μg / mL (2 μg / mL to 200 μg / mL) were labeled as (C).

[1151]

[1152] Example 48: Comparative Study 3. Plasma Protein Binding Protocol Summary

[1153]

[1154]

[1155] Test Procedure

[1156] The test compound was incorporated into plasma to a final concentration of 10 μM. 300 μL plasma aliquots were placed in the red chamber of the insert, and 500 μL PBS was placed in the white chamber of the insert. The plate was incubated at 37 °C in a thermomixer at 400 rpm for 5 hours. After incubation, the samples were matrix-matched with the opposite matrix (10 μL plasma / 100 μL buffer sample was matched with 100 μL blank buffer / 10 μL plasma). The matrix-matched samples were precipitated with 200 μL of acetonitrile containing 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 LC-MS / MS. Blank control samples were processed immediately after preparation of the plasma working stock solution. These samples were used as a measure to calculate the % recovery of the test compound.

[1157] Data analysis and calculation

[1158] The percentage of plasma binding fraction was calculated by the following equation:

[1159] Unbound % = 100 * F C / T C

[1160] Recovery % = 100 * (F C +T C ) / T 0

[1161] where

[1162] T C = total compound concentration determined by the calculated concentration on the plasma side of the membrane

[1163] F C = free compound concentration determined by the calculated concentration on the buffer side of the membrane

[1164] T 0 = total compound concentration determined before dialysis

[1165] For each set of replicates / compounds, the percentage, unbound percentage and recovery percentage were determined. The results are shown below.

[1166] 4. Plasma stability of the test compound

[1167] Protocol summary

[1168]

[1169]

[1170] Test procedure

[1171] Incubate the test compound and QC compound at a final concentration of 1 μM in plasma in a shaking water bath at 37 °C with gentle shaking. At the predetermined time points, terminate the reaction with 200 μL of acetonitrile containing an internal standard and centrifuge at 4000 × RCF at 4 °C for 20 minutes. Separate the supernatant and analyze it by LC-MS / MS.

[1172] Data analysis and calculation

[1173] Use the following equation to determine the percentage of the test compound / QC compound remaining after the above procedure:

[1174]

[1175] The results are shown below.

[1176] Studies were conducted to compare the compounds of the present invention with structurally similar compounds ("Control Example X"). The experiments were carried out as described above. As shown below, the data were segmented.

[1177] For VIM-1, a Ki value < 0.15 μM is labeled (++++); a Ki value of 0.15 μM to 0.3 μM is labeled (+++); a Ki value of 0.3 μM to 0.5 μM is labeled (++); a Ki value > 0.5 μM (0.5 μM to 1 μM) is labeled (+).

[1178] For IMP-1, a Ki value < 0.15 μM is labeled (++++); a Ki value of 0.15 μM to 0.6 μM is labeled (+++); a Ki value of 0.6 μM to 5 μM is labeled (++); a Ki value > 5 μM (5 to 10 μM) is labeled (+).

[1179] For VIM-2, a Ki value < 0.02 μM is labeled (++++); a Ki value of 0.02 μM to 0.05 μM is labeled (+++); a Ki value of 0.05 μM to 0.1 μM is labeled (++); a Ki value > 0.1 μM (0.1 to 0.15 μM) is labeled (+).

[1180] For NDM-1, a Ki value < 0.03 μM is labeled (++++); a Ki value of 0.03 μM to 0.1 μM is labeled (+++); a Ki value of 0.1 to 0.3 is labeled (++); a Ki value > 0.5 μM (0.3 μM to 2 μM) is labeled (+).

[1181]

[1182]

[1183] Observe the MBL inhibitory potency of the compound of Example 7.

[1184]

[1185]

[1186]

[1187]

[1188] ND: Undetermined

[1189] Analogues related to the structure lacking the -C(NR)-NR 2 motif show that the compounds of the present invention exhibit better enzyme inhibition (lower Ki values) and better potentiation (lower MIC values) against the above-mentioned MBL enzymes (VIM / IMP / NDM) and bacterial strains than the analogues.

[1190] It can be readily seen that the compounds of the present invention are associated with improved properties compared to structurally similar compounds. This finding is surprising, especially since the -C(NR)-NR 2 motif commonly found in the compounds of the present invention may be associated with rapid hydrolysis and it might have been expected that the compounds would not be suitable for the uses described herein. Thus, the fact that the introduction of this motif not only does not compromise the efficacy of the compounds but also improves plasma stability and efficacy is unexpected.

[1191] 5. PK (Pharmacokinetics) studies.

[1192] Compound A and Example 7 were intravenously administered to male Swiss albino mice at 1 mg / kg. The measured PK parameters are shown in the following table:

[1193] Compound <![CDATA[C 0 (ng / mL)]]> AUC (ng.h / mL) Cl (mL / min / kg) Compound A 17 9 819 Example 7 1814 1036 16

[1194] C 0 = Plasma concentration

[1195] AUC = Area under the curve

[1196] Cl = Clearance

[1197] Note that for the same dose:

[1198] 1. Compared to Compound A, Example 7 achieves a maximum concentration more than 100-fold higher;

[1199] 2. Compared to Compound A, Example 7 achieves more than 100-fold higher exposure (AUC, integral of concentration versus time); and

[1200] 3. Example 7 is cleared from the blood approximately 50-fold slower than Compound A.

[1201] This data is consistent with the in vitro data generated on plasma stability and confirms plasma stability as a limiting factor in terms of the potential of Compound A available for in vivo efficacy studies.

[1202] 6. In Vivo Efficacy Studies

[1203] The thighs of mice were infected with Klebsiella pneumoniae NTBC104. Since this strain produces NDM-1, the MIC of meropenem against this strain is 64 μg / mL. In the presence of 8 μg / mL of the compound of Example 2, the MIC of meropenem is 4 μg / mL.

[1204] At the end of the experiment (9 hours after infection), the animals were sacrificed and the number of colony-forming units (CFUs) was measured to quantify the bacterial load (degree of infection). 30 mg / kg of meropenem slightly reduced the bacterial load, while 30 mg / kg of meropenem plus 30 mg / kg of the compound of Example 2 showed a 1.6 Log reduction in CFUs compared to meropenem alone 10 , significantly reducing the bacterial load. The results are as Figure 1 shown.

[1205] Under the same experimental conditions, the compound of Example 7 achieved a 1.7 Log reduction in CFUs compared to meropenem alone 10 . No efficacy study was conducted on Compound A because in vitro and PK studies predicted that this compound would fail in efficacy studies, and thus it would be unethical to conduct such an experiment.

[1206] Under the same conditions, the compound of Example 26 achieved a 1.8 Log reduction in CFUs compared to meropenem alone 10 .

[1207] 7. Extended MIC Analysis with Clinical Strains Expressing MBL

[1208] To evaluate the coverage of the compounds of the present invention and the enhancing effect on meropenem, the susceptibilities of approximately 200 clinical isolates were examined. The criteria for inclusion in the panel were that the clinical strains were resistant to carbapenems but expressed only NDM enzyme variants and did not express serine β-lactamases with carbapenemase activity such as KPC or OXA.

[1209] At a concentration of 8 μg / mL of Example 2 or Example 26, meropenem was enhanced to the extent that nearly 90% of the strains showed a meropenem MIC of 8 μg / mL, while the concentration of meropenem alone only stopped the growth of <1% of the strains, and within the parameters of this experiment, meropenem alone could not achieve 90% growth arrest of all strains. The results are as Figure 2 shown.

[1210] Example 49

[1211] 1. Combination therapy of the compounds of the present invention with SBL inhibitors and antibiotic agents

[1212] As described above, bacteria exhibit resistance to antibiotics through the following mechanisms: including altering biological targets to reduce the binding affinity for antibiotics, and producing enzymes that inactivate antibacterial drugs, such as β-lactamases (including serine-β-lactamases (SBLs) and metallo-β-lactamases (MBLs)). A proposed strategy to address this resistance is to administer combination therapy that includes an agent that inhibits the enzyme that inactivates the antibiotic and the antibiotic itself. In other words, the antibacterial activity of the drug can be rescued by using a dual combination approach of an antibiotic plus a drug that inhibits the inactivating enzyme.

[1213] The combination of a serine β-lactamase inhibitor and an antibiotic is known. For example, the Streptomyces natural product clavulanic acid (a serine β-lactamase inhibitor) was developed in combination with the β-lactam antibiotic amoxicillin as the dual combination known as Augmentin. More recently, the combination of avibactam (a serine β-lactamase inhibitor with an improved serine β-lactamase inhibition profile compared to clavulanic acid) and the cephalosporin β-lactam antibiotic ceftazidime (collectively known as Avycaz) has been introduced clinically. However, these combinations are ineffective in treating bacterial infections caused by bacteria expressing MBL enzymes because SBL inhibitors are generally inactive against these enzymes.

[1214] Due to the lack of a diagnostic test that can very rapidly determine the exact mechanism of β-lactamase resistance currently available clinically, and also due to the lack of a dual inhibitor of serine β-lactamases and metallo-β-lactamases, there is a further complication of the two different classes of β-lactamases present in bacterial infections. In fact, even if a rapid diagnostic test were available to distinguish resistance due to SBL enzymes from resistance due to MBL enzymes, there is currently no clinically approved metallo-β-lactamase inhibitor to address the problem of the presence of metallo-β-lactamases.

[1215] The inventors have now recognized that products of drug combinations that are antibiotics, serine β-lactamase inhibitors, and metallo-β-lactamase inhibitors (the so-called triple combination) can overcome the need to identify whether the resistant bacteria causing a particular infection produce 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 the β-lactamase profiles of carbapenem-resistant Enterobacteriaceae (CRE). Group 1 organisms have only metallo-β-lactamase or a mixture of metallo-β-lactamase and serine β-lactamase, but resistance is mainly attributed to metallo-β-lactamase. Group 2 organisms have only serine β-lactamase. Group 3 organisms have both metallo-β-lactamase and serine β-lactamase, and both enzymes play an important role in resistance.

[1216] The following abbreviations are used in this example:

[1217] CMY: Class C β-lactamase

[1218] TEM: Class A β-lactamase

[1219] SHV: Class A B-lactamase (sulfhydryl variable)

[1220] CTX-M: Class A β-lactamase (CTX stands for cefotaximase and M stands for Munich)

[1221] OSBL: "older-spectrum" β-lactamase

[1222] OXA: Class D β-lactamase (oxacillinase)

[1223] ACT-TYPE: Class C β-lactamase (AmpC-type β-lactamase)

[1224] KPC: Class A β-lactamase (Klebsiella pneumoniae carbapenemase)

[1225] VIM: Verona integron-encoded metallo-β-lactamase

[1226] NDM: New Delhi metallo-β-lactamase

[1227] IMP: Imipenemase metallo-β-lactamase

[1228] The experiment was carried out using the "broth microdilution method" according to protocol M07-A8 determined by the Clinical and Laboratory Standards Institute (CLSI). Serial dilutions of meropenem (mero) were prepared in 96-well plates in cation-adjusted Mueller-Hinton broth (CAMHB); the concentration range was limited to 0.03 mg / L to 512 mg / L. The compounds (the compound of Example 2 above and / or WCK4234) were added at the concentrations shown in the following table. The bacterial inoculum of each strain (clinical isolate) was adjusted to a McFarland turbidity standard of 0.5 in physiological serum (0.9% NaCl), then diluted 1:100 in CAMHB and added to each well to obtain a final bacterial cell count of 5×10 5 CFU / well. After incubation in a heating chamber at 37 °C for 18 to 20 hours, growth inhibition was evaluated based on the absence of any bacterial amplification.

[1229] The minimum inhibitory concentration (MIC) was taken as the lowest antibiotic concentration at which the test organism did not show visible growth; the results were confirmed by measuring the optical density (OD) at 600 nm in a spectrophotometer.

[1230] The SBL inhibitor WCK4234 was synthesized according to the method described in WO 2015 / 114595.

[1231]

[1232] M: H (WCK4234) or Na (sodium salt of WCK4234)

[1233] Briefly, WCK4234 and its sodium salt were synthesized according to the disclosed method (WO2105114595), and the subsequent stages are shown below:

[1234]

[1235] The compound of formula (B) was prepared by the synthetic method described in detail by Ball, M. et al in Organic Process Research and Development, (2016), 1799.

[1236] The clinical strains used in these experiments were as follows:

[1237] Group 1 (strains with resistance mainly attributed to metallo-β-lactamases):

[1238] NTBC020 (Escherichia coli strain expressing NDM-1, TEM-1, and CTX-M-15), NTBC035-2 (Klebsiella pneumoniae strain expressing NDM-1, CMY-4, and SHV-11), NTBC104-1 (Klebsiella pneumoniae strain expressing NDM-1 and SHV-11), NTBC123 (Klebsiella pneumoniae strain expressing NDM-1), NTBC062 (Klebsiella pneumoniae strain expressing IMP-1 and TEM-1), NTBC024 (Klebsiella pneumoniae strain expressing VIM-19, TEM-1, and CTX-M-3), NTBC042 (Escherichia coli strain expressing VIM-1, TEM-1, CTX-M-15, SHV-12), NTBC055 (Escherichia coli strain expressing VIM-1), and NTBC039 (Klebsiella oxytoca strain expressing IMP-28).

[1239] Group 2 (strains with resistance attributed to serine β-lactamases):

[1240] NTBC091-1 (Escherichia coli strain expressing KPC-2 and TEM-1), NTBC093 (E. cloacae strain expressing KPC-2 and TEM-1), NTBC096-1 (Klebsiella pneumoniae strain expressing OXA-181 and SHV-11), NTBC099 (Klebsiella pneumoniae strain expressing KPC-3, SHV-11, and TEM-1), and NTBC189 (Klebsiella pneumoniae strain expressing TEM-OSBL, CTX-M-14, and OXA-48).

[1241] Group 3 (strains with resistance attributed to serine β-lactamases and metallo-β-lactamases):

[1242] NTBC019 (Klebsiella pneumoniae strain expressing NDM-1, CTX-M-15, and OXA-181), NTBC185 (Klebsiella pneumoniae strain expressing SHV-OSBL, TEM-OSBL, NDM-1, and OXA-48), NTBC186 (Klebsiella pneumoniae strain expressing ACT-TYPE, VIM-1, and OXA-48), NTBC187 (Klebsiella pneumoniae strain expressing SHV-OSBL, NDM-1, and OXA-48), and NTBC188 (Klebsiella pneumoniae strain expressing NDM-1 and KPC-2).

[1243] The results are as follows. The data are segmented as follows: MIC values < 1 μg / mL are labeled as (A); MIC values of 1 μg / mL or 2 μg / mL are labeled as (B); MIC values of 4 μg / mL or 8 μg / mL are labeled as (C); MIC values ≥ 16 μg / mL are labeled as (D).

[1244]

[1245] It can be seen from the strains of Group 1 and Group 2 that the dual combination of meropenem and a suitable β-lactamase inhibitor reduces the MIC required.

[1246] For the organisms of Group 3, the results show that the combination of the MBL inhibitor and the SBL inhibitor (WCK4234) according to the invention and meropenem is capable of reducing the MIC required.

[1247] Discussion

[1248] In the field of antibacterials, there are no known examples of triple therapies specifically for eradicating bacterial infections. There is one example of a triple therapy for treating gastroesophageal reflux disease (GORD) where Helicobacter pylori (H. pylori) infection is suspected to be a component of the disease and gastric ulcers, but in this case, the National Institute of Clinical Excellence (NICE) guidelines recommend treatment with a triple combination of an anti-ulcer proton pump inhibitor and two antibiotics (amoxicillin and clarithromycin). There are no triple combinations of antibacterials or antibacterials plus adjuvants such as β-lactamase inhibitors either in development or in clinical use.

[1249] A significant advantage provided by the triple combination of the present invention is that when encountering CRE strains and rapid treatment is necessary for the patient to survive, using the triple combination means that in principle there is no need to wait for microbiological and molecular characterization of the resistance elements before starting treatment. Thus, the triple combination described herein can be used to prevent or treat any bacterial infection as it obviates the need for prior identification of the bacterial strain.

[1250] 2. Further data

[1251] Additional experiments were conducted to demonstrate the advantages of the triple combination of the present invention.

[1252] The experiments were conducted as described above. The compounds at 8 μg / mL (the compounds of Examples 2 and 26) were tested. Avibactam and Wck4234 at 4 μg / mL were tested. The following MIC values were determined:

[1253] Test strains that simultaneously express a carbapenemase from class B (MBL) and a carbapenemase from class A or D (serine β-lactamase).

[1254] NTBC19 is Klebsiella pneumoniae expressing NDM-1, CTXM-15, and OXA-181.

[1255] NTBC188 is Enterobacter cloacae expressing NDM-1 and KPC-2.

[1256] The data were segmented as follows: MIC values ≤ 0.5 μg / mL were labeled (A); MIC values of 1 μg / mL to 4 μg / mL were labeled (B); MIC values ≥ 8 μg / mL (8 μg / mL to 512 μg / mL) were labeled (C). The results are shown in the table below.

[1257]

[1258]

[1259] The data clearly show that the triple combination of (i) meropenem, (ii) the compounds of the present invention (e.g., the compound of Example 2 or the compound of Example 26), and (iii) an SBL inhibitor (e.g., avibactam or WCK4234) advantageously results in a decrease in the MIC values of the two tested strains.

Claims

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein, ○R 1 is H; ○ is a cyclic group selected from C 6 to C 10 aryl and 5- to 10-membered heteroaryl containing 1, 2 or 3 heteroatoms selected from O, S or N; ○ Each R 2 is independently selected from: (i) Halogen or R 8 , where each R 8 is independently selected from CN and C(O)NR f R g ; where each R f and R g is H; and (ii)O(C 1-3 alkyl), which may optionally be substituted by three halogen substituents; ○ m is 0, 1, 2 or 3 ○R 3 is hydrogen; ○ 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 C 1-3 an alkylene group; or L is -C(R 10 )=N-; ○ X is a bond, or when L is not a bond or -C(R 10 )=N-, 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 unsubstituted or -NR 10 R 11 substituted C 1 to C 3 alkyl; or R 4 linked to R 5 are joined together with the atoms to which they are attached to form a 5- or 6-membered heterocyclic group which contains at least one saturated carbon atom in the ring and contains one or two heteroatoms selected from O, S and N; and (ii) R 6 selected from: H, -CN, and unsubstituted or -NR 10 R 11 substituted C 1 to C 3 alkyl; and, if R 7 is present, selected from H, -CN, and C 1 to C 3 alkyl; or if R 7 is present, then R 6 is linked to R 7 together with the atoms to which they are linked to form a 5- to 6-membered heterocyclic group containing at least one saturated carbon atom and containing 1 or 2 heteroatoms selected from O, S and N in the ring; or B -R 4 is H; -R 5 linked to R 6 are linked together to form, together with the atoms to which they are linked, a 5- or 6-membered heterocyclic group containing at least one saturated carbon atom in the ring and containing 1 or 2 heteroatoms selected from O, S and N; and -R 7 If present, selected from: H, -CN and C 1 to C 3 alkyl; ○ Each R 10 and R 11 is H.

2. The compound according to claim 1, wherein, is a cyclic group selected from phenyl and 5- to 6-membered heteroaryl containing 1, 2 or 3 heteroatoms selected from O, S and N.

3. The compound according to claim 2, wherein, Selected from phenyl, pyridazine, pyridine and thiazole.

4. The compound according to claim 1, wherein, is phenyl.

5. The compound according to claim 1, wherein, Each R 2 is independently selected from: (i) Halogen, CN, OH, -OCF 3 , -OCH 3 and -C(O)NR f R g ; wherein R f and R g is H.

6. The compound according to claim 1, wherein, Each R 2 is fluorine.

7. The compound according to claim 1, wherein, n is 0.

8. The compound according to claim 1, wherein, Z is a bond or selected from -NR 10 C(O)-, -C(O)NR 10 -, and -NR 10 C(O)NR 11 .

9. The compound according to claim 1, wherein, Z is selected from -NR 10 C(O)-, -C(O)NR 10 -, and -NR 10 C(O)NR 11 -.

10. The compound according to claim 1, wherein, L is a bond.

11. The compound according to claim 1, wherein L is C 1-3 alkylene.

12. The compound according to claim 1, wherein, X is a bond.

13. The compound according to claim 1, wherein, p is 1; and R 7 is H or methyl.

14. The compound according to claim 1, wherein, R 4 is H.

15. The compound according to claim 1, wherein, R 5 is H and R 6 is H or methyl.

16. The compound according to claim 1, wherein: ●R 1 is H; ● is phenyl; ● m is 2; ● Each R 2 is independently a halogen; ● n is 0; ●Z is selected from -NR 10 C(O)-, -C(O)NR 10 -, and -NR 10 C(O)NR 11 -; ●L is a key or C 1-3 an alkylene group; ● X is a bond; ●p is 1; and R 4 is H; R 5 is selected from H, -CN and C 1 to C 2 alkyl; R 6 is H or methyl, and R 7 is H or methyl.

17. The compound according to claim 1, wherein ·R 1 is H; · is phenyl; · m is 2; · Each R 2 is fluorine; · n is 0; ·Z is -NR 10 C(O)NR 11 -; · L is a bond; · X is a bond; ·p is 1 and R 7 is H; ·R 4 is H; ·R 5 is H; and ·R 6 is H.

18. The compound according to claim 1, wherein, R 4 、R 5 、R 6 and R 7 each is hydrogen if present.

19. The compound according to claim 1, the compound is 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-guanidinoethylthiocarbonylamino)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-Formylhydrazino)-2-oxo-ethyl]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylic acid; ● 5-[[3-Chloro-4-[(2-guanidinoacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylic acid; ● 5-[[4-[(2-Guanidinoacetyl)amino]-3-methoxy-phenyl]sulfonylamino]thiazole-4-carboxylic acid; ● 5-[[4-[[2-(2-formamidinohydrazino)acetyl]amino]-3-fluorophenyl]sulfonylamino]thiazole-4-carboxylic acid; ● 5-[[4-[[(2E)-2-formamidinohydrazono)acetyl]amino]-3-fluorophenyl]sulfonylamino]thiazole-4-carboxylic acid; ● 5-[[4-[[2-(4,5-dihydro-1H-imidazol-2-ylamino)acetyl]amino]-3,5-difluorophenyl]sulfonylamino]thiazole-4-carboxylic acid; ● 5-[[6-[(2-guanidinoacetyl)amino]pyridazin-3-yl]sulfonylamino]thiazole-4-carboxylic acid; ● 5-[[4-[(2-amino-2-iminoethyl)carbamoylamino]-3-fluorophenyl]sulfonylamino]thiazole-4-carboxylic acid; · 5-[[3,5-difluoro-4-(guanidinoaminocarbonylamino)phenyl]sulfonylamino]thiazole-4-carboxylic acid; ● 5-[[4-[(3-amino-3-imino-propionyl)amino]-3,5-difluorophenyl]sulfonylamino]thiazole-4-carboxylic acid; ● 5-[[4-[[3-(dimethylamino)-3-imino-propionyl]amino]-3-fluorophenyl]sulfonylamino]thiazole-4-carboxylic acid; ● 5-[[3-fluoro-4-[(2-guanidinooxyacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylic acid; ● 5-[[3-fluoro-4-[[3-imino-3-(methylamino)propionyl]amino]phenyl]sulfonylamino]thiazole-4-carboxylic acid; ● 5-[[4-[3-(4,5-dihydro-1H-imidazol-2-yl)propionylamino]-3-fluorophenyl]sulfonylamino]thiazole-4-carboxylic acid; ● 5-[[2-[(2-guanidinoacetyl)amino]thiazol-5-yl]sulfonylamino]thiazole-4-carboxylic acid; ● 5-[[4-[[2-[(N-cyanomethylamidine)amino]acetyl]amino]-3-fluorophenyl]sulfonylamino]thiazole-4-carboxylic acid; ● 5-[[3-fluoro-4-(guanidinoaminocarbonylamino)phenyl]sulfonylamino]thiazole-4-carboxylic acid; ● 5-[[3-fluoro-4-[[2-(morpholine-4-carboxamidoamino)acetyl]amino]phenyl]sulfonylamino]thiazole-4-carboxylic acid; ● 5-[[4-[(3-amino-3-imino-2-methyl-propionyl)amino]-3-fluorophenyl]sulfonylamino]thiazole-4-carboxylic acid; ● 5-[[4-[[2-(4,5-dihydro-1H-imidazol-2-yl)acetyl]amino]-3-fluorophenyl]sulfonylamino]thiazole-4-carboxylic acid; ● 5-[[4-(formamidinocarbonylamino)-3-fluorophenyl]sulfonylamino]thiazole-4-carboxylic acid; ● 5-[[4-[[(2R)-2-guanidinopropionyl]amino]phenyl]sulfonylamino]thiazole-4-carboxylic acid; ● 5-[[3,5-difluoro-4-[(2-guanidinoacetyl)amino]phenyl]sulfonylamino]thiazole-4-carboxylic acid; ● 5-[[4-[(4-amino-4-imino-butanoyl)amino]-3-fluoro-phenyl]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-[(N-methylformamidinyl)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-[formamido(methyl)amino]acetyl]amino]-3-fluoro-phenyl]sulfonylamino]thiazole-4-carboxylic acid; ● 5-[[4-[[2-[[N-(2-aminoethyl)formamidinyl]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-guanidinopropionylamino)phenyl]sulfonylamino]thiazole-4-carboxylic acid; ● 5-[[4-[(3-amino-3-imino-propionyl)amino]-3-fluoro-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.

20. A pharmaceutical composition comprising a compound according to any one of claims 1 to 19 and at least one pharmaceutically acceptable carrier or diluent.

21. The pharmaceutical composition according to claim 20, wherein, the pharmaceutical composition further comprises (i) an antibiotic agent and / or (ii) a serine-β-lactamase inhibitor.

22. The pharmaceutical composition according to claim 21, wherein, the antibiotic agent is a β-lactam antibiotic.

23. The pharmaceutical composition according to claim 22, wherein, the β-lactam antibiotic is selected from carbapenems, penicillins, cephalosporins and penems.

24. The pharmaceutical composition according to claim 23, wherein, the β-lactam antibiotic is meropenem.

25. The pharmaceutical composition according to claim 21, wherein, A: the serine-β-lactamase inhibitor is a compound of formula (II) or its pharmaceutically acceptable salt, wherein, ○G is selected from -CN and -C(O)NR j R k ; ○R k Selected from -W- and -Q-W; wherein W is selected from 5- to 6-membered heterocyclic groups, R j and -N(R j ) 2 ; and Q is selected from -NR j C(O)-, -C(O)-NR j -, C 1-3 alkylene, -O-C 1-3 alkylene and -N(R j )-C 1-3 alkylene; ○ Each R j is selected from H and unsubstituted C 1-3 alkyl or B: the serine-β-lactamase inhibitor is selected from WCK4234, avibactam, relebactam, zidebactam and nacubactam or their pharmaceutically acceptable salts.

26. Use of a compound according to any one of claims 1 to 19 in the manufacture of a medicament for removing or reducing antibiotic resistance in Gram-negative bacteria.

27. Use according to claim 26, wherein, the Gram-negative bacteria are selected from the Enterobacteriaceae, Pseudomonadaceae and Moraxellaceae.

28. Use according to claim 27, wherein, the Gram-negative bacteria are selected from Klebsiella pneumoniae, Escherichia coli, Pseudomonas aeruginosa, Burkholderia cepacia and Acinetobacter baumannii.

29. Use of a compound according to any one of claims 1 to 19 in the preparation of a medicament for treating or preventing a bacterial infection.

30. Use according to claim 29, wherein, the bacterial infection is caused by bacteria selected from the Enterobacteriaceae, Pseudomonadaceae and Moraxellaceae.

31. Use according to claim 30, wherein, the bacteria selected from the Enterobacteriaceae, Pseudomonadaceae and Moraxellaceae are selected from Klebsiella pneumoniae, Escherichia coli, Pseudomonas aeruginosa, Burkholderia cepacia and Acinetobacter baumannii.

32. Use according to claim 29, wherein, the bacterial infection is caused by carbapenem-resistant Enterobacteriaceae.

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