Thiadiazolone derivatives useful as AMPK activators

Abstract

The present invention relates to a compound of formula I, or a pharma- ceutically acceptable salt or solvate thereof, 1 , R 2 , R 3 and R 4 are as defined herein, and these compounds are useful, particularly as prodrugs, in the treatment of disorders or conditions ameliorated by activation of AMPK.

Description

Detailed description of the invention

[0001] [Field of Invention] This invention relates to novel compounds and their use in pharmaceuticals. In particular, the invention relates to compounds useful for treating disorders or health conditions that are improved by the activation of AMP-activated protein kinase.

[0002] [Background of the Invention] AMP-activated protein kinase (AMPK) is a protein kinase enzyme composed of three protein subunits that is activated by hormones, cytokines, exercise, and stress that reduces the cellular energy state (e.g., glucose deficiency). AMPK activation increases the process that produces adenosine 5'-triphosphate (ATP) (e.g., fatty acid oxidation) and suppresses other processes that consume ATP but are not rapidly required for survival, such as fatty acid, glycerolipid, and protein synthesis. Conversely, when cells are presented with a sustained excess of glucose, AMPK activity decreases and fatty acid, glycerolipid, and protein synthesis are enhanced. Therefore, AMPK is a protein kinase enzyme that plays a crucial role in cellular energy homeostasis. Consequently, AMPK activation, coupled with the glucose-lowering effect, induces several other biological effects, including inhibition of cholesterol synthesis, lipid production, triglyceride synthesis, and reduction of hyperinsulinemia.

[0003] Considering the above, AMPK is a preferred target for the treatment of metabolic syndrome, particularly type 2 diabetes. AMPK is also involved in many pathways that are important for a wide variety of diseases (for example, AMPK is also involved in many pathways that are important in CNS disorders, fibrosis, osteoporosis, heart failure, and sexual dysfunction).

[0004] AMPK is also involved in many important pathways in cancer. Several tumor suppressors are part of the AMPK pathway. AMPK acts as a negative regulator of the mammalian TOR (mTOR) and EF2 pathways, which are important regulators of cell growth and proliferation. Therefore, deregulation can be associated with diseases such as cancer (and diabetes). Thus, AMPK activators may be useful as anticancer agents.

[0005] AMPK activators (e.g., metformin and 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide (i.e., the compound of formula II below)) have been shown to be effective in treating pain. Das and collaborators reported that post-injury treatment with AMPK activators in mice after lumbar disc puncture reduced mechanical hypersensitivity (Das V, et al. Reg Anesth Pain Med 2019;0:1-5. doi:10.1136 / rapm-2019-100839). Similarly, Das and his collaborators have reported that early treatment with AMPK activators reduces mechanical hypersensitivity in a mouse model of postoperative pain (Das V, et al. Reg Anesth Pain Med 2019;0:1-6. doi:10.1136 / rapm-2019-100651). These drugs also normalize the AMPK pathway in the dorsal root ganglia. Therefore, AMPK activators can be used to treat pain, particularly postoperative pain.

[0006] It has also been shown that hepatic steatopathies can be regulated by AMPK (Zhao et al. J. Biol. Chem. 2020 295: 12279-12289). AMPK activation promotes fatty acid oxidation (β-oxidation) in the liver while inhibiting de novo lipid production. AMPK activation also reduces the release of free fatty acids from adipose tissue, thus preventing hepatic steatopathies. Pharmacological activation of AMPK in the liver has been reported to promote beneficial effects on multiple aspects of non-alcoholic fatty liver disease (NAFLD). For example, AMPK activation has been found to improve non-alcoholic steatohepatitis (NASH) in both mouse and monkey animal models. Therefore, AMPK activators may be useful in the treatment of NAFLD and NASH.

[0007] An example of an AMPK activator is 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide (i.e., the compound of formula II), which was first disclosed in WO2011 / 004162. [ka]

[0008] As AMPK agonists (i.e., AMPK activators), compounds of formula II are useful in treating disorders or health conditions that are improved by AMPK activation. These compounds may be useful in treating cardiovascular diseases (such as heart failure), diabetic nephropathy, type 2 diabetes, insulin resistance, non-alcoholic fatty liver disease, non-alcoholic steatohepatitis, pain, opioid dependence, obesity, cancer, inflammation (including chronic inflammatory diseases), autoimmune diseases, osteoporosis, and intestinal diseases.

[0009] Many AMPK activators are known, but a need still exists for the development of novel compounds for the treatment of disorders or health conditions ameliorated by activation of AMPK. The present inventors have now found novel compounds that are metabolized in vivo to form known AMPK activators, resulting in a surprisingly improved bioavailability of the AMPK activators. The compounds were also found to activate AMPK.

[0010] Any list or discussion of a document that is clearly published previously in this specification should not necessarily be construed as an admission that the document is part of the prior art or constitutes common general knowledge.

[0011] [Detailed Description of the Invention] In a first aspect of the present invention, there is provided a compound of Formula I, or a pharmaceutically acceptable salt or solvate thereof: [Chemical Formula] R 1 and R 2 are independently selected from the group consisting of hydrogen, the side chains of proteinogenic amino acids, and C optionally substituted with one or more substituents selected from the group consisting of -OH, -NH₂ and -C(O)NH₂ 1-6 alkyl; R 3 and R 4 are independently selected from the group consisting of hydrogen, C optionally substituted with a phenyl group 1-4 alkyl, and A 1 ; or R 1 (or R 2 ) and R 3 (or R 4 ) together with the carbon atom and nitrogen atom to which they are attached form a 4- to 6-membered heterocycloalkyl group; A 1 is [Chemical Formula] or [ka] This represents, A 2 C is optionally substituted with hydrogen or a phenyl group. 1-4 Alkyl, [ka] or [ka] This represents, Each A 3 This is independently a C that is optionally substituted with hydrogen or a phenyl group. 1-4 Represents alkyl; and R 5a and R 5b This is independently a C molecule optionally substituted with hydrogen, a side chain of a protein constituent amino acid, and one or more substituents selected from the group consisting of -OH, -NH2, and -C(O)NH2. 1-6 Selected from the group consisting of alkyl groups.

[0012] These compounds, including their pharmaceutically acceptable salts and solvates, may be referred to herein as “compounds of the present invention.”

[0013] In a preferred embodiment of the first aspect of the present invention, A 2 is hydrogen, [ka] or [ka] This represents, Each A 3 This represents hydrogen.

[0014] The compounds of the present invention have been found to be metabolized in vivo to form 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide (referred to herein as the compound of formula II), which is known as an AMPK activator. In this respect, the compounds of the present invention can be considered to be prodrugs of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide.

[0015] Potentially useful pharmaceutically acceptable salts include those discussed by Berge et al. in J. Pharmaceutical Sciences, 66: 1-19 (1977). Pharmacopoeia-acceptable salts of the compound of formula I may also be prepared according to techniques well known to those skilled in the art.

[0016] Examples of pharmaceutically acceptable addition salts include addition salts derived from organic acids such as citric acid, tartaric acid, acetic acid (including halogenated forms of acetic acid such as trifluoroacetic acid), malic acid, lactic acid, fumaric acid, benzoic acid, glycolic acid, gluconic acid, succinic acid, and arylsulfonic acid; as well as addition salts derived from mineral acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, metaphosphoric acid, nitric acid, and sulfuric acid. In certain embodiments, the pharmaceutically acceptable salt is the citrate or trifluoroacetate salt of the compound of formula I.

[0017] Unless otherwise specified, alkyl groups as defined herein may be linear, branched, and / or cyclic (thus forming a cycloalkyl group) if a sufficient number of carbon atoms (i.e., at least two or three as required) are present. If a sufficient number of carbon atoms (i.e., at least four) are present, the group may also be partially cyclic (thus forming a partial cycloalkyl group). For example, cycloalkyl groups that may be listed include cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. Similarly, partial cyclic alkyl groups that may be listed (these may also be referred to as “partially cycloalkyl” groups) include cyclopropylmethyl.

[0018] The heterocycloalkyl groups that may be listed include non-aromatic monocyclic heterocycloalkyl groups in which at least one (e.g., 1 to 4) atoms in the ring system is a non-carbon atom (i.e., a heteroatom, e.g., sulfur, oxygen, or especially nitrogen), and the total number of atoms in the ring system is 4 to 6. The carbon atoms of the heterocycloalkyl groups referred to herein may be substituted by one or more =O substituents.

[0019] To avoid any ambiguity, even if two or more substituents in a compound of formula I are identical in identity, the actual identity of each substituent is not dependent on any one other.

[0020] Where a group is referred to as optionally substituted in this specification, it is particularly intended that such optional substituent may be absent (i.e., the reference to such optional substituent may be omitted), in which case the optionally substituted group may be referred to as unsubstituted in certain embodiments.

[0021] A pharmaceutically acceptable salt of the compound of formula I may be prepared according to the art well known to those skilled in the art. For example, the compound of formula I may be reacted with a suitable organic or mineral acid. Salt switching techniques may be used to convert one salt to another.

[0022] The compounds disclosed herein may exist in both solvated and non-solvated forms, as well as in solvated forms with pharmaceutically acceptable solvents such as water and ethanol, and the present invention is intended to encompass both solvated and non-solvated forms of the compounds of the present invention.

[0023] The term "solvate" refers to a stoichiometrically altered complex formed by a solute and a solvent. For the purposes of this invention, the solvent does not need to interfere with the biological activity of the solute. Examples of suitable solvents include, but are not limited to, water, methanol, ethanol, and acetic acid. Solvates in which water is the solvent molecule are typically referred to as hydrates. Hydrates include compositions containing a stoichiometric amount of water and compositions containing a variable amount of water.

[0024] Since the compounds of formula I contain double bonds, they may exist as E (entgegen) and Z (zusammen) geometric isomers for each individual double bond. All such isomers and mixtures thereof are included within the scope of the present invention.

[0025] The compound of formula I may exist as a positional isomer or may exhibit tautomerism. All tautomers and mixtures thereof are included within the scope of the present invention.

[0026] The present invention also includes isotope-labeled compounds of formula I, identical to those described herein, due to the fact that one or more atoms are substituted with atoms having atomic masses or mass numbers different from those commonly found in nature (or the most abundant atomic masses or mass numbers found in nature). All isotopes of any particular atom or element defined herein are intended to be within the scope of the present invention. Accordingly, compounds of formula I also include deuterated compounds, i.e., compounds of formula I in which one or more hydrogen atoms are substituted with hydrogen isotopes.

[0027] Those skilled in the art will understand that the compounds of the present invention, which are the subject of this invention, include stable compounds. That is, the compounds of the present invention include compounds that are robust enough to be isolated, for example, from a reaction mixture to a useful purity.

[0028] Throughout this specification, structures may or may not be indicated by their chemical names. In case of any doubt regarding nomenclature, the structure shall prevail. Where a compound may exist as a tautomer (e.g., another resonance form), the structure shown represents one of the possible tautomers, and the tautomer actually observed may vary depending on environmental factors such as solvent, temperature, or pH. All tautomer (and resonance) forms, as well as mixtures thereof, are included within the scope of this invention.

[0029] Unless otherwise indicated, all technical and scientific terms used herein have the general meanings understood by those skilled in the art to which this invention pertains.

[0030] To avoid any doubt, those skilled in the art will understand that any reference herein to a particular aspect of the present invention (e.g., a first aspect of the present invention) includes references to all of its embodiments and specific features, and that these embodiments and specific features may be used in combination to form further embodiments and features of the present invention.

[0031] Amino acids are organic compounds containing an amino functional group (-NH2) and a carboxyl functional group (-CO2H), and (in most cases) have side chains. α-amino acids are amino acids in which the amino functional group and the carboxyl functional group are bonded to the same carbon atom (i.e., the α-carbon atom). Among the α-amino acids that may be listed, protein-constituent amino acids are particularly noteworthy. Protein-constituent amino acids are amino acids that are biosynthetically incorporated into proteins during translation. The amino acids that make up proteins are glycine (Gly), alanine (Ala), arginine (Arg), asparagine (Asn), aspartic acid (Asp), cysteine ​​(Cys), glutamic acid (Glu), glutamine (Gln), histidine (His), isoleucine (Ile), leucine (Leu), lysine (Lys), methionine (Met), phenylalanine (Phe), proline (Pro), pyrrolicin (Pyl), selenocysteine ​​(Sec), serine (Ser), threonine (Thr), tryptophan (Trp), tyrosine (Tyr), and valine (Val).

[0032] Therefore, the term "side chain of a protein-constituting amino acid" refers to a group other than hydrogen bonded to the α-carbon atom of a protein-constituting amino acid. In the context of the present invention, the side chain of one or more protein-constituting amino acids is the R of the compound of formula I. 1 , R 2 , R 5a or R 5b It may be located at any of these positions. To avoid doubt, the compounds of the present invention may include multiple different side chains at these positions, i.e., the prodrug of compound II may include a chain having two or three different amino acids linked to the thiadiazole ring. Specific examples of protein-constituting amino acid side chains include methyl substituted with an imidazolyl or indolyl group (for histidine and tryptophan), propyl substituted with a guanidino group (for arginine), and butyl substituted with an amino group (for lysine).

[0033] Specific compounds of the present invention that may be listed are R 1 and R2 These are compounds in which R is identical. For example, R 1 and R 2 Both can be hydrogen.

[0034] Other compounds of the present invention that may be listed are R 1 and R 2 These are different compounds. For example, R 1 These are the side chains of protein constituent amino acids, and R 2 It may be hydrogen.

[0035] Therefore, the compounds of the first aspect of the present invention may contain one or more asymmetric carbon atoms and thus may exhibit optical isomerism and / or diastereoisomerism. In particular, R 1 and R 2 (For example, R 1 is methyl, R 2 The compound of formula I (where is hydrogen) is, as will be understood by those skilled in the art, R 1 and R 2 The compound may have a carbon atom substituted with R in an L-configuration or a D-configuration. 5a and / or R 5b If it contains a group, then their R 5a and R 5b The chiral center to which the group is attached may also independently be in either an L-configuration or a D-configuration.

[0036] In a particular embodiment, R 1 and R 2 If R is different, 1 and R 2 The carbon substituted with is in the L- configuration. Alternatively, R 1 and R 2 If R is different, 1 and R 2 The substituted carbon may be in a D-configuration. For example, the compound of formula I may be the compound of formula IA or the compound of formula I. [ka] R 1 , R 2 , R 3 and R 4 This is as described herein (i.e., as described in the First Aspect of the Invention, including all embodiments and specific features, and combinations thereof). Specific examples of the compounds of the Invention are R 1 and R 2 Both are hydrogen, or R 1 and R 2 A compound in which the substituted carbon is in an L-configuration, for example, R 1 and R 2 When one of the atoms is hydrogen, the other is a side chain of a protein-constituting amino acid.

[0037] Similarly, R 3 or R 4 is A 1 When representing A 1 teeth [ka] It may also represent A 2 C is optionally substituted with hydrogen or a phenyl group. 1-4 Alkyl, [ka] It may also represent A 3 , R 5a and R 5b This is as described herein (i.e., as described in the First Aspect of the Invention, including all embodiments and specific features, and combinations thereof). Specific examples of the compounds of the Invention are A 1 This is a compound that represents a protein-constituting amino acid or two linked protein-constituting amino acids.

[0038] Preferred examples of the compounds of the present invention that may be listed include A 2 hydrogen, [ka] or [ka] Represents; each A 3 represents hydrogen; and R 5b Examples of compounds are as described herein.

[0039] Preferably, R 1 C is optionally substituted with hydrogen, the side chain of a protein constituent amino acid, and one or more substituents selected from the group consisting of -OH, -NH2 and -C(O)NH2. 1-6 Selected from the group consisting of alkyl groups, or R 1 and R 3 They are linked together to form a pyrrolidine ring (found in proline); and R 2 It is hydrogen.

[0040] In a particular embodiment, R 2 is hydrogen, R 1 R is a side chain of a protein constituent amino acid selected from the group consisting of hydrogen, or Glu, particularly Arg, His, Lys, Ser, Thr, Asn, Gln, Cys, Sec, Ala, Ile, Leu, Met, Phe, Trp, Tyr, Asp, and Val; or R 2 is hydrogen, R 1 and R 3 These, along with the carbon and nitrogen atoms to which they are bonded, form a pyrrolidine ring.

[0041] In a particular embodiment, R 2 is hydrogen, and R 1 R is a hydrogen atom or a side chain of an amino acid selected from the group consisting of Arg, His, Lys, and Trp (e.g., Arg, His, and Lys); or R 2 is hydrogen, R 1 and R 3together with the carbon atom and nitrogen atom to which they are bonded form a pyrrolidine ring.

[0042] In a further embodiment, R 2 is hydrogen, and R 1 is hydrogen, or a side chain of an amino acid selected from the group consisting of Lys, Ala, Ile, Phe, Leu, Ser and Asp; alternatively, R 2 is hydrogen, and R 1 and R 3 together with the carbon atom and nitrogen atom to which they are bonded form a pyrrolidine ring.

[0043] Preferably, R 2 is hydrogen.

[0044] Certain embodiments of the present invention include compounds of Formula I, wherein R 1 is hydrogen, or a side chain of an amino acid (e.g., Arg, His and Lys) selected from the group consisting of Arg, His, Lys and Trp; alternatively, R 1 and R 3 together with the carbon atom and nitrogen atom to which they are bonded form a pyrrolidine ring; and R 2 is hydrogen.

[0045] In other embodiments, R 3 and R 4 are independently hydrogen, C optionally substituted by one or more phenyl groups (e.g., methyl group or benzyl group) 1-3 alkyl, [Formula] and [Formula] selected from the group consisting of; and R 5a is hydrogen or a side chain of a proteinogenic amino acid. In a preferred embodiment, R 3 and R4 C is independently optionally substituted with hydrogen and one or more phenyl groups. 1-3 Selected from the group consisting of alkyl groups (for example, R 3 and R 4 (is independently hydrogen, methyl, or benzyl). Preferably, R 3 and R 4 R is independently selected from the group consisting of hydrogen and methyl (for example, R 3 and R 4 (Both are methyl.)

[0046] In another preferred embodiment, R 3 and R 4 C is independently a hydrogen atom, optionally substituted with one or more phenyl groups (e.g., a methyl group or a benzyl group). 1-3 Alkyl, [ka] and [ka] Selected from the group consisting of; or, R 1 and R 3 They form a pyrrolidine ring together with the carbon and nitrogen atoms to which they are bonded, R 4 C is optionally substituted with hydrogen and one or more phenyl groups. 1-3 Alkyl, [ka] and [ka] Selected from the group consisting of; and R 5a These are the side chains of protein-constituting amino acids (e.g., Phe, Val, Lys, Asp, or Arg).

[0047] Preferred compounds of the present invention that may be listed include R 1 However, hydrogen, or Glu, is a side chain of a protein constituent amino acid selected from the group consisting of Arg, His, Lys, Ser, Thr, Asn, Gln, Cys, Sec, Ala, Ile, Leu, Met, Phe, Trp, Tyr, Asp, and Val (e.g., Lys, Ala, Ile, Phe, Leu, Ser, and Asp; or Arg, His, Lys, and Trp); R 2 R represents hydrogen; 3 and R 4 C independently, optionally substituted with hydrogen and one or more phenyl groups (e.g., methyl). 1-3 Selected from the group consisting of alkyl groups; or R 1 and R 3 However, these compounds include those that form a pyrrolidine ring with the carbon and nitrogen atoms to which they are bonded.

[0048] Other preferred compounds of the present invention that may be listed include R 2 R represents hydrogen; 1 R is a side chain of a protein constituent amino acid selected from the group consisting of hydrogen, or Glu, Arg, His, Lys, Ser, Thr, Asn, Gln, Cys, Sec, Ala, Ile, Leu, Met, Phe, Trp, Tyr, Asp and Val (e.g., Lys, Ala, Ile, Phe, Leu, Ser and Asp); 3 and R 4 C independently, optionally substituted with hydrogen, one or more phenyl groups (e.g., methyl or benzyl groups). 1-3 Alkyl, [ka] and [ka] Selected from the group consisting of; or, R 1 and R 3However, together with the carbon and nitrogen atoms to which they are bonded, they form a pyrrolidine ring, R 4 However, C is optionally substituted with hydrogen, one or more phenyl groups (e.g., a methyl group or a benzyl group). 1-3 Alkyl, [ka] and [ka] Selected from the group consisting of; R 5a but is a side chain of a protein constituent amino acid selected from the group consisting of hydrogen, or Glu, Arg, His, Lys, Ser, Thr, Asn, Gln, Cys, Sec, Ala, Ile, Leu, Met, Phe, Trp, Tyr, Asp and Val (e.g., Phe, Val, Lys, Asp and Arg); and A 2 and A 3 This includes compounds that are independently defined as herein.

[0049] Embodiments of the present invention, A 2 teeth [ka] Represents R 5b Preferably, is a side chain of a protein constituent amino acid selected from the group consisting of hydrogen or Glu, Arg, His, Lys, Ser, Thr, Asn, Gln, Cys, Sec, Ala, Ile, Leu, Met, Phe, Trp, Tyr, Asp and Val (e.g., Phe, Val, Lys, Asp and Arg); and each A 3 Hydrogen and C are independent of each other. 1-3 Selected from the group consisting of alkyl (e.g., methyl).

[0050] As described above, the compounds of the present invention have been found to be metabolized in vivo to form 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide (referred to herein as the compound of formula II). Nevertheless, the exact mechanism by which the compounds of the present invention are converted to the compound of formula II may vary depending on the compound. While we do not wish to be bound by theory, it is thought that the compounds of the present invention are broken down to the compound of formula II in two steps. First, the ester is hydrolyzed (chemically or enzymatically) to form an N-hydroxymethyl intermediate (referred to elsewhere herein as the compound of formula III), and then this intermediate is converted to the compound of formula II.

[0051] The applicant found that some compounds are hydrolyzed more rapidly under acidic conditions (e.g., conditions expected to be present in the human stomach), while others are hydrolyzed more rapidly under more alkaline conditions (e.g., conditions similar to those in the small intestine).

[0052] The compounds of the present invention, which are hydrolyzed more rapidly under alkaline conditions (e.g., conditions similar to those in the small intestine) than under acidic conditions, are advantageous in that they may offer further clinical benefits, for example, they may be useful in patients who cannot achieve sufficient conversion in the stomach (e.g., cannot achieve conversion at the required rate or to the required extent). Such patients may, for example, suffer from esterase enzyme dysfunction. Furthermore, the intermediates formed as a result of hydrolysis (i.e., compounds of formula III) may be more readily taken up in the intestines compared to compounds of formula II, and therefore, the formation of compounds of formula III in the intestines may be advantageous in achieving optimal systemic exposure for the patient.

[0053] R 1 / R 2 The compound of the present invention, which contains a lipophilic (alkyl) group, was found to have relatively high stability at pH 7.4. 1 / R 2When the group is an H or -NH2-containing amino acid side chain (e.g., glycine-based or lysine-based), its stability at this pH is low. 1 / R 2 When one of the groups is an electron-withdrawing amino acid side chain (e.g., an aspartic acid-based, arginine-based, or glutamic acid-based side chain), the stability at this pH is very low.

[0054] Specific compounds that may be listed in this regard are the compounds of formula A or their pharmaceutically acceptable salts or solvates: [ka] (i)R 1 represents hydrogen, R 2 R represents the side chain of aspartic acid, the side chain of arginine, or the side chain of glutamic acid. 3 R is either hydrogen or methyl, 4 is hydrogen or methyl; or (ii)R 1 , R 2 and R 3 represents hydrogen, R 4 is A 1 This represents A 1 teeth [ka] This represents A 2 is hydrogen or methyl, A 3 R is either hydrogen or methyl, 5a This represents the side chain of aspartic acid, the side chain of arginine, or the side chain of glutamic acid.

[0055] Further compounds that may be listed in this regard are the compounds of formula B or their pharmaceutically acceptable salts or solvates: [ka] (i)R 1 represents hydrogen, R 2represents the side chain of aspartic acid or the side chain of arginine, R 3 R is either hydrogen or methyl, 4 is hydrogen or methyl; or (ii)R 1 , R 2 and R 3 represents hydrogen, R 4 is A 1 This represents A 1 teeth [ka] This represents A 2 is hydrogen or methyl, A 3 R is either hydrogen or methyl, 5a This represents the side chain of aspartic acid or the side chain of arginine.

[0056] Further specific compounds that may be listed in this regard are compounds of formula C or their pharmaceutically acceptable salts or solvates: [ka] (i)R 1 , R 3 and R 4 represents hydrogen, R 2 represents the side chain of aspartic acid, the side chain of arginine, or the side chain of glutamic acid; or (ii)R 1 , R 2 and R 3 represents hydrogen, R 4 is A 1 This represents A 1 teeth [ka] Represents R 5a This represents the side chain of aspartic acid, the side chain of arginine, or the side chain of glutamic acid.

[0057] In another embodiment, the compound of the present invention is R 2It is hydrogen, and furthermore, R 1 The side chain is aspartic acid, and optionally, R 3 and R 4 It is a compound of formula I, wherein the atoms are independently hydrogen or methyl.

[0058] In this regard, further compounds that may be listed are the following compounds and their pharmaceutically acceptable salts or solvates: [ka]

[0059] In further embodiments, the present invention also relates to compounds of formula I as defined herein (including compounds of formula I as defined according to any of the preferred embodiments specified herein), provided that: (i) The compound is not the compound of formula A; or (ii) The compound is not a compound of formula B; or (iii) The compound is not a compound of formula C; or (iv) The compound is not selected from the following group: [ka] or (v) Compound R 2 is hydrogen, R 1 The side chain of aspartic acid is (optionally, R 3 and R 4 Not a compound of formula I (where each is independently hydrogen or methyl); or (compared to) compounds [ka] isn't it.

[0060] Particularly preferred compounds of the present invention are the following compounds or their pharmaceutically acceptable salts or solvates. [ka] [ka] [ka]

[0061] Since the compound of the present invention has been found to decompose in vivo to form the above compound, it can be considered a prodrug of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide. 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide can be represented as the compound of formula II. [ka]

[0062] The term "prodrug" refers to a compound that, after oral or parenteral administration, is metabolized in vivo to produce a pharmaceutically active compound in experimentally detectable amounts within a specified time frame (e.g., within a 6-24 hour dosing interval, i.e., 1-4 times per day). To avoid ambiguity, the term "parenteral" administration includes all forms of administration other than oral administration. General information on prodrugs can be found, for example, in Bundegaard, H. "Design of Prodrugs" p. l-92, Elsevier, New York-Oxford (1985).

[0063] Surprisingly, administration of the compound of the present invention resulted in a significant increase in the bioavailability and systemic exposure of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide compared to the bioavailability and systemic exposure observed after administration of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide.

[0064] The bioavailability of a drug is the amount of the administered dose that reaches the systemic circulation in its original form. Sufficient bioavailability is important for achieving a therapeutically effective concentration at the site of action. An improvement (i.e., increase) in bioavailability is the amount of the drug that reaches the blood C15 after administration of the compound (or its pharmaceutical formulation) to the subject. max Alternatively, this may be demonstrated by measuring the area under the curve (AUC). The compounds and formulations of the present invention are useful for therapies in subjects requiring the therapies described herein. Subjects that may be listed include animals such as mammals. Specific mammals that may be listed include, for example, primates (e.g., humans, male or female), cattle, horses, dogs, and cats. Preferably, the subject is human.

[0065] The term “C max Those skilled in the art will understand that in this context, "AUC" refers to the peak plasma concentration of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide after administration (for example, to human subjects), and the integral of the concentration / time curve of the compound (or formulation thereof) of the present invention after administration, respectively.

[0066] As is evident from the data from the examples, administration of the compound of the present invention significantly improved the bioavailability of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide in vivo. These data indicate that administration of the compound of the present invention to mammalian subjects increases plasma exposure to 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide compared to plasma exposure observed after administration of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide.

[0067] Therefore, administration of the compound of the present invention can increase the bioavailability of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide compared to administration of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide. In this context, the expression "increases bioavailability" means that administration of the compound of the present invention results in a larger systemically available fraction of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide in vivo compared to administration of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide. The increase in the systemically available fraction of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide may be at least about 10%, (at least) about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100% (i.e., 2x), about 200% (i.e., 3x), about 300% (i.e., 4x), about 400% (i.e., 5x), about 500% (i.e., 6x), about 600% (i.e., 7x), about 700% (i.e., 8x), about 800% (i.e., 9x), or about 900% (i.e., 10x).

[0068] The improved bioavailability provided by the compounds of the present invention may be demonstrated using preferred methods known in the art. For example, the improved bioavailability may be demonstrated by comparing pharmacokinetic data (e.g., AUC data) of subjects administered with the compounds of the present invention with corresponding data of subjects administered with 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide.

[0069] As shown in the examples, the compounds of the present invention exhibit improved stability under acidic conditions (e.g., pH 1.2). Therefore, the compounds of the present invention are suitable for the preparation of pharmaceutical formulations for oral administration to patients requiring gastric passage.

[0070] The compounds of the present invention have also been found to be remarkably effective in activating AMPK, as demonstrated by the data from the examples. As AMPK activators (i.e., AMPK agonists), the compounds of the present invention may be useful in treating disorders or health conditions that are improved by AMPK activation. Accordingly, the compounds may be useful in treating the specific diseases described herein.

[0071] The compounds of the present invention may be prepared from 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide. 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide may be prepared sequentially according to techniques well known to those skilled in the art. For example, 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide may be produced according to the technique described in international patent application WO2011 / 004162, the details of which are incorporated herein by reference.

[0072] The compounds of the present invention described herein may be prepared according to techniques well known to those skilled in the art, such as the techniques described in the examples provided below.

[0073] Compounds of formula I may be obtained from readily available starting materials using appropriate reagents and reaction conditions, by analogy with methods known in the literature or by conventional synthetic procedures, in accordance with standard techniques. In this regard, those skilled in the art may refer in particular to "Comprehensive Organic Synthesis" by BM Trost and I. Fleming, Pergamon Press, 1991.

[0074] For example, a method for preparing the compounds of the present invention as defined herein is provided. (i) Compounds of formula III, [ka] Reaction with the compound of formula IV [ka] or (ii) Compound of formula V, [ka] This includes reactions with the compound of formula IV, [ka] R 1 and R 2 R is as defined herein. 3 and R 4 The process is carried out in the presence of a suitable protecting group (e.g., Boc), as defined herein, or independently, in the presence of a suitable base (e.g., 4-dimethylaminopyridine), a suitable amide coupling agent (e.g., N,N'-dicyclohexylcarbodiimide), and a suitable solvent (e.g., tetrahydrofuran), according to procedures known to those skilled in the art.

[0075] To avoid any doubt, the methods described herein for preparing the compounds of the present invention include methods comprising a series of reactions between a compound of formula III and up to three (e.g., two) compounds of formula IV. For example, the following steps are included: (i) A step of reacting the compound of formula III with the compound of formula IV according to the method described herein; (ii) Optionally, a step in which one or more deprotection steps are performed; (iii) A step of reacting the product of step (i) or step (ii) with the compound of formula IV, according to the method described herein; (iv) Optionally, a step of performing one or more deprotection steps; (v) optionally, a step of reacting the product of step (iii) or step (iv) with the compound of formula IV in accordance with the method described herein; and (vi) A step in which one or more deprotection steps are optionally performed.

[0076] In the above embodiment, the compound of formula IV may be the same or different in each of steps (i), (iii), and (v).

[0077] Those skilled in the art will understand that in the methods described above and below, it may be necessary to protect the functional groups of the intermediate compound with protecting groups. For example, in certain embodiments, in the compound of formula IV, R 3 and R 4 One of the groups may represent hydrogen, and the other may represent a suitable protecting group (e.g., Boc). Protection and deprotection of the functional group may be carried out before or after the above reaction.

[0078] Protecting groups are well known to those skilled in the art and may be removed by techniques such as those described below. For example, the protected compounds / intermediates described herein may be chemically converted to unprotected compounds using standard deprotection techniques. The use of protecting groups is fully described in “Protective Groups in Organic Synthesis”, 3rd edition, TW Greene & PGM Wutz, Wiley-Interscience (1999).

[0079] Therefore, specific transformation steps that may be employed to form the compound of formula I include a deprotection step, for example, deprotection of the N-Boc protecting group by reaction in the presence of an acid, or a hydroxyl group protected as a silyl ether (e.g., a tert-butyl-dimethylsilyl protecting group) may be deprotected by reaction with an acid or fluoride ion source, for example, by the use of the reagent tetrabutylammonium fluoride (TBAF).

[0080] The compound of formula III may be obtained from available starting materials using standard techniques and conventional synthetic procedures, with appropriate reagents and reaction conditions. For example, the compound of formula III (referred to as compound 2 in the examples) may be prepared by reacting the compound of formula II with formaldehyde in the presence of a suitable base (e.g., triethylamine) and a suitable solvent (e.g., N,N-dimethylformamide), according to procedures known to those skilled in the art.

[0081] Similarly, the compound of formula II (i.e., 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide; referred to as compound 1 in the examples) may be prepared according to techniques well known to those skilled in the art, for example, the techniques described in international patent application no. WO2011 / 004162.

[0082] Those skilled in the art will recognize that compounds of formula V may be obtained from readily available starting materials by the synthetic procedures illustrated herein, using appropriate reagents and reaction conditions.

[0083] The compounds of the present invention may be isolated from the reaction mixture and, if necessary, purified using prior art known to those skilled in the art. Accordingly, the methods for preparing the compounds of the present invention described herein may include, as a final step, isolation and optionally purification of the compounds of the present invention.

[0084] [Pharmaceutical preparations] As described herein, the compounds of the present invention are useful as therapeutic agents for treating various medical disorders or health conditions. Typically, the compounds of the present invention are administered to subjects in need in the form of pharmaceutical formulations.

[0085] According to a second aspect of the present invention, a pharmaceutical formulation comprising a compound of formula I (or a pharmaceutically acceptable salt or solvate thereof) is provided. This formulation is referred to herein as the formulation of the present invention. All embodiments and their specific features described herein in relation to the first aspect of the present invention are disclosed herein in relation to the second aspect of the present invention.

[0086] A pharmaceutical formulation according to a second aspect of the present invention may be prepared in accordance with standard and / or recognized pharmaceutical practice.

[0087] In a second embodiment of the present invention, the compound of the present invention (or a pharmaceutically acceptable salt or solvate thereof) is the sole pharmaceutically active ingredient present in the formulation. In a further embodiment of the second embodiment of the present invention, the compound of the present invention (or a pharmaceutically acceptable salt or solvate thereof) may be present in the formulation together with one or more other pharmaceutically active ingredients, or administered as part of a combination therapy with one or more other pharmaceutically active ingredients.

[0088] Formulations according to a second aspect of the present invention are generally provided as a mixture comprising the compound of the present invention (or a pharmaceutically acceptable salt or solvate thereof) and one or more pharmaceutically acceptable excipients. The one or more pharmaceutically acceptable excipients may be selected in accordance with standard pharmaceutical practice, with appropriate consideration given to the intended route of administration. The pharmaceutically acceptable excipients are preferably chemically inert to the active compound and preferably do not have adverse side effects or toxicity under the conditions of use. Suitable pharmaceutical formulations may be found, for example, in Remington, The Science and Practice of Pharmacy, 19th ed., Mack Printing Company, Easton, Pennsylvania (1995). A brief overview of drug delivery methods may be found, for example, in Langer, Science 249, 1527 (1990).

[0089] For example, the formulation of the present invention may contain lubricants, binders, fillers, surfactants, diluents, anti-adhesion agents, coatings, flavoring agents, colorants, flow accelerators, preservatives, sweeteners, disintegrants, adsorbents, buffers, antioxidants, chelating agents, dissolution accelerators, dissolution retarders, and / or wetting agents.

[0090] Certain pharmaceutically acceptable excipients that may be listed include microcrystalline cellulose, lactose monohydrate, crospovidone, magnesium stearate, colloidal silicon dioxide, anhydrous lactose, dicalcium phosphate, mannitol, pregelatinized starch, hydroxypropylcellulose, povidone, low-substituted hydroxypropylcellulose, croscarmellose sodium, sodium starch glycolate, sodium stearyl fumarate, talc, hydroxypropyl methylcellulose, polysorbate 80, sodium lauryl sulfate, poloxamer 188, poloxamer 407, propylene glycol, titanium dioxide, hypromellose phthalate, hypromellose acetate succinate, methacrylic acid, methyl methacrylate copolymer, Opadry® I, and Opadry® II.

[0091] In the preparation of pharmaceutical formulations of the compound of the present invention for oral administration, the compound of the present invention may be mixed together or separately with one or more of the above-mentioned pharmaceutical excipients.

[0092] A mixture of the compound of the present invention and one or more pharmaceutically acceptable excipients may be processed into pellets or granules, or compressed into tablets. Accordingly, the pharmaceutical formulations of the present invention include formulations provided in the form of tablets, minitablets, blocks, pellets, particles, granules, or powders for oral administration. The mixture of the compound of the present invention and one or more pharmaceutically acceptable excipients may also be provided in a form suitable for subcutaneous or intramuscular administration, such as an injection solution or a lyophilized powder suitable for reconstitution in a suitable fluid before administration.

[0093] Those skilled in the art will understand that the formulations described herein may act systemically and may therefore be administered using preferred techniques known to those skilled in the art. The formulations described herein are typically administered orally, subcutaneously, or intramuscularly in a preferred pharmaceutically acceptable dosage form. Pharmaceutical formulations of a second aspect of the present invention are preferably oral pharmaceutical formulations, particularly in terms of improving the stability under acidic conditions observed for the compounds of the present invention.

[0094] The formulations of the present invention may be prepared for oral administration in the form of capsules. For example, capsules such as soft gelatin capsules may be prepared to contain the compound of the present invention (or a pharmaceutically acceptable salt or solvate thereof) alone, together with an optionally suitable vehicle, such as vegetable oil or fat. Similarly, hard gelatin capsules may contain the compound of the present invention (or a pharmaceutically acceptable salt or solvate thereof) alone, or in combination with solid powder components such as disaccharides (e.g., lactose or saccharose), alcoholic sugars (e.g., sorbitol or mannitol), vegetable starches (e.g., potato starch or corn starch), polysaccharides (e.g., amylopectin or cellulose derivatives), or gelling agents (e.g., gelatin).

[0095] The specific pharmaceutical formulations of the present invention may include formulations that are provided, for example, in the form of capsules or tablets for oral administration.

[0096] Pharmaceutical formulations that may be listed include those in which the compound of the present invention (or a pharmaceutically acceptable salt or solvate thereof) is present in an amount of at least 1% by weight (or at least 10% by weight, at least 30% by weight, or at least 50% by weight) and up to 99% by weight of the formulation. The weight ratio of the compound of the present invention (or a pharmaceutically acceptable salt or solvate thereof) to the total components of the pharmaceutical formulation (i.e., the compound of the present invention and all pharmaceutical excipients, e.g., adjuvants, diluents and carriers) is at least 1:99 (or at least 10:90, at least 30:70, or at least 50:50) and up to 99:1.

[0097] As used herein, “therapeutably effective amount,” “effective amount,” and “dosage” refer to the amount of the compound of the present invention (or a pharmaceutically acceptable salt or solvate thereof) sufficient to produce a desired clinical effect that may be a therapeutic and / or beneficial effect. The effective amount or dosage varies depending on the age or general health of the subject (e.g., human), the severity of the health condition being treated, the specific drug administered, the duration of treatment, the nature of any concurrent treatment, the pharmaceutically acceptable excipients used, and similar factors within the scope of the knowledge and expertise of those skilled in the art. Appropriately, the “therapeutably effective amount,” “effective amount,” or “dosage” in individual cases may be determined by those skilled in the art by referring to relevant texts and literature and / or by using routine experiments. Those skilled in the art will understand that the therapeutic effect does not need to be complete or curative, as long as some benefit is provided to the subject.

[0098] Those skilled in the art will understand that the compounds of the present invention and their formulations may be administered in various doses (for example, by one or more preparations described herein), and that a suitable dose can be easily determined by those skilled in the art. The total dose of the compounds of the present invention (or their pharmaceutically acceptable salts or solvates) administered to the target subject may range from about 1 milligram / day (mg / day) to about 3000 mg / day, about 2 mg / day to about 2000 mg / day, or about 5 mg / day to about 1000 mg / day (for example, about 10 mg / day to about 500 mg / day). Such dose may be, for example, the oral dose of a formulation according to a second embodiment of the present invention. When administering the compounds of the present invention (or their pharmaceutically acceptable salts or solvates) intramuscularly or subcutaneously, those skilled in the art will recognize that the dose should be adjusted as appropriate.

[0099] When administered orally, treatment with the above formulation may include administration of a unit-dose formulation containing about 1 mg to about 3000 mg of the compound of the present invention, for example, about 2 mg to about 2000 mg, or about 5 mg to about 1000 mg (for example, about 10 mg to about 500 mg) of the compound of the present invention. Advantageously, treatment may include administration of the compound of the present invention (for example, in the form of one or more capsules containing the above formulation) using a once-daily dose. Alternatively, the total daily dose of the compound of the present invention may be administered in two, three, or four divided doses per day (for example, twice daily, referring to the doses described herein, for example, doses of 100 mg, 250 mg, 500 mg, or 1000 mg twice daily). A specialist will recognize that the dosage will vary depending on the subject.

[0100] In certain embodiments, the daily dose of the compound of the present invention administered to a subject is in the range of about 1 mg to about 3000 mg, preferably about 1 mg to about 1000 mg.

[0101] As used herein, the term "about" refers to a variation of 20%, 10%, 5%, 1%, 0.5%, or even 0.1% of a particular quantity when referring to measurable values ​​such as the amount, dose, time, or temperature of a compound. In each case, it is intended that the term may be replaced with notation such as "±10%" (or by indicating the variance of a particular quantity calculated based on the relevant value). It is also intended that the term may be deleted in each case.

[0102] To avoid any doubt, in the context of the present invention, the dose administered to a subject, particularly a human subject, should be sufficient to produce a therapeutic response in the subject over a reasonable time frame. Those skilled in the art will recognize that the precise dose and composition, as well as the selection of the most appropriate delivery regimen, are influenced, among other things, by the pharmacological properties of the formulation, the nature and severity of the health condition being treated, the physical and mental sensitivities of the recipient, the potency of the particular compound, the age, condition, weight, sex and response of the subject being treated, and the stage / severity of the disease.

[0103] In any case, physicians or other persons skilled in the art will be able to routinely determine the most appropriate actual dosage for individual subjects. The above dosages are examples of average cases; of course, there may be individual cases where higher or lower dosage ranges are appropriate, and such cases are also within the scope of the present invention.

[0104] [Medical use] As described herein, the compounds of the present invention are useful as pharmaceuticals. The compounds of the present invention are useful because they have pharmacological activity and / or are metabolized in the body after oral or parenteral administration to form compounds that have pharmacological activity.

[0105] Accordingly, according to a third aspect of the present invention, pharmaceutical formulations are provided for use in pharmaceuticals, which include compounds of the present invention as already defined herein (i.e., compounds defined in the first aspect of the present invention) or pharmaceutical formulations defined in relation to a second aspect of the present invention. To avoid doubt, references to compounds defined in the first aspect of the present invention include references to compounds of formula I (including all embodiments thereof) and their pharmaceutically acceptable salts and solvates.

[0106] The compounds of the present invention (i.e., the compounds defined in the first aspect of the present invention) and formulations containing the same may be particularly useful in treating disorders or health conditions that are improved by the activation of AMP-activated protein kinase (AMPK). Accordingly, in a fourth aspect of the present invention, the compounds of the present invention, or formulations containing the compound, are provided for use in treating disorders or health conditions that are improved by the activation of AMPK.

[0107] Similarly, the use of the compounds of the present invention, or formulations containing such compounds, in the manufacture of pharmaceuticals for the treatment of disorders or health conditions that are improved by activation of AMPK is provided. In a further alternative fourth embodiment of the present invention, a method is provided for treating a disorder or health condition that is improved by activation of AMPK, comprising administering the compounds of the present invention (or formulations containing such compounds) to a subject (e.g., a human) that requires it.

[0108] "Activating AMPK" means that the steady-state level of phosphorylation of the Thr-172 portion of the AMPK-α (AMPK-alpha) subunit increases compared to the steady-state level of phosphorylation in the absence of the compound of formula I or its active metabolite (e.g., 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide). Alternatively, it means that a higher steady-state level of phosphorylation of any other protein downstream of AMPK, such as acetyl-CoA carboxylase (ACC), is present.

[0109] Those skilled in the art will understand that the term “a disorder or health condition improved by AMPK activation” includes cardiovascular disease (e.g., heart failure), diabetic nephropathy, diabetes (e.g., type 2 diabetes), insulin resistance, non-alcoholic fatty liver disease, non-alcoholic steatohepatitis, pain, opioid dependence, obesity, cancer, inflammation (including chronic inflammatory diseases), autoimmune diseases, osteoporosis, intestinal diseases, and hyperinsulinemia associated with obesity or cardiovascular disease. Other diseases or health conditions that may be improved by AMPK activation include hyperinsulinemia and related conditions, health conditions / disorders in which fibrosis plays a role, sexual dysfunction, and neurodegenerative diseases.

[0110] Those skilled in the art will understand that the term “cancer” encompasses one or more diseases in a type of disorder characterized by the uncontrolled division of cells and their ability to invade other tissues, either by direct growth into adjacent tissues through invasion and proliferation, or by transplantation to distant sites through metastasis. “Proliferation” includes an increase in the number and / or size of cancer cells. “Metastasis” means the movement or migration (e.g., invasion) of cancer cells from a primary tumor site within the body of the subject to one or more other areas within the body of the subject (the cells may then form secondary tumors).

[0111] Therefore, the compounds of the present invention may be suitable for use in the treatment of any type of cancer, including all tumors (non-solid and preferably solid tumors, e.g., carcinomas, adenomas, adenocarcinomas, blood cancers, regardless of organ). For example, cancer cells may be selected from the group consisting of cancer cells of the breast, bile duct, brain, colon, stomach, genitals, thyroid, hematopoietic system, lungs and airways, skin, gallbladder, liver, nasopharynx, nerve cells, kidneys, prostate, lymph nodes and gastrointestinal tract. Preferably, cancer is selected from the group consisting of colon cancer (including colorectal adenoma), breast cancer (e.g., postmenopausal breast cancer), endometrial cancer, hematopoietic system cancers (e.g., leukemia, lymphoma, etc.), thyroid cancer, kidney cancer, esophageal adenocarcinoma, ovarian cancer, prostate cancer, pancreatic cancer, gallbladder cancer, liver cancer and cervical cancer. More preferably, cancer is selected from the group consisting of colon, prostate, and especially breast cancer. When the cancer is a non-solid tumor, it is preferably a hematopoietic malignancy such as leukemia (e.g., acute myeloid leukemia (AML), chronic myeloid leukemia (CML), acute lymphoblastic leukemia (ALL), or chronic lymphocytic leukemia (CLL)). Preferably, the cancer cells are breast cancer cells.

[0112] Those skilled in the art will understand that the term “diabetes” (i.e., diabetes mellitus) refers to both type 1 (insulin-dependent) diabetes mellitus and type 2 (insulin-independent) diabetes mellitus, both of which involve dysfunction of glucose homeostasis. The compounds of the present invention and their formulations may be particularly suitable for use in the treatment of type 1 diabetes mellitus and / or type 2 diabetes mellitus. The compounds of the present invention are particularly suitable for the treatment of type 2 diabetes mellitus.

[0113] In addition to being useful in the treatment of diabetes, the compounds of the present invention are also suitable for the treatment of diabetic nephropathy (i.e., diabetic nephropathy). "Diabetic nephropathy" refers to kidney damage caused by diabetes and is a serious complication of type 1 and type 2 diabetes. Diabetic nephropathy affects the kidney's ability to remove waste products from the blood and excrete them as urine, which can lead to renal failure.

[0114] The compounds of the present invention are also suitable for treating chronic kidney disease, including chronic kidney disease in the absence of type 2 diabetes. "Chronic kidney disease" is a health condition characterized by a progressive loss of kidney function over time. Chronic kidney disease usually results from one or more other diseases or health conditions affecting the kidneys, such as hypertension, diabetes, high cholesterol, kidney infections, glomerulonephritis, polycystic kidney disease, obstruction of the urinary tract in the flow of urine, and long-term drug use.

[0115] It is understood by those skilled in the art that the term “hyperinsulinemia or related conditions” includes hyperinsulinemia, type 2 diabetes, glucose intolerance, insulin resistance, metabolic syndrome, dyslipidemia, childhood hyperinsulinemia, hypercholesterolemia, hypertension, obesity, fatty liver conditions, diabetic nephropathy, diabetic neuropathy, diabetic retinopathy, cardiovascular disease, atherosclerosis, cerebrovascular conditions such as stroke, neurodegenerative diseases such as systemic lupus erythematosus and Alzheimer's disease, and polycystic ovary syndrome. Other disease conditions include progressive renal diseases such as chronic renal failure.

[0116] In particular, the compounds and formulations of the present invention may be suitable for use in the treatment of obesity associated with hyperinsulinemia and / or cardiovascular disease associated with hyperinsulinemia.

[0117] The compounds and formulations of the present invention may also be suitable for use in the treatment of cardiovascular diseases (e.g., heart failure) that are not associated with hyperinsulinemia. Similarly, the compounds and formulations of the present invention may also be suitable for use in the treatment of obesity that is not associated with hyperinsulinemia. To avoid doubt, the treatment of obesity and / or cardiovascular diseases (such as heart failure) in which AMPK activation may be beneficial is included within the scope of the present invention.

[0118] Health conditions / disorders in which fibrosis plays a role include, but are not limited to, scar healing, keloids, scleroderma, pulmonary fibrosis (including idiopathic pulmonary fibrosis), nephrogenic systemic fibrosis, and cardiovascular fibrosis (including endocardial myofibrosis), systemic sclerosis, cirrhosis, macular degeneration, retinal and vitreoretinopathy, Crohn's disease / inflammatory bowel disease, surgical scar formation, radiation and chemotherapy-induced fibrosis, and cardiovascular fibrosis.

[0119] The compounds of the present invention may also be useful in treating sexual dysfunction (e.g., erectile dysfunction). The compounds of the present invention may also be useful in treating inflammation.

[0120] Neurodegenerative diseases that may be listed include Alzheimer's disease, Parkinson's disease and Huntington's disease, polyglutamine disorders such as amyotrophic lateral sclerosis, spinal and ocular muscle atrophy (SBMA), dentate nucleus and pallidoluysian atrophy (DRPLA), and some spinocerebellar ataxias (SCAs).

[0121] The compounds of the present invention may be useful in the treatment of non-alcoholic fatty liver disease (NAFLD).

[0122] Non-alcoholic fatty liver disease (NAFLD) is defined by excessive fat accumulation in the liver in the form of triglycerides (steatosis) (histologically referred to as accumulation of more than 5% of hepatocytes). It is the most common liver disorder in developed countries (for example, affecting about 30% of adults in the United States), and most patients are asymptomatic. If left untreated, the condition can gradually worsen and eventually lead to cirrhosis. NAFLD is particularly common in obese individuals, with an estimated 80% having the disease.

[0123] NAFLD may be diagnosed when alcohol consumption is not considered the primary causative factor in the patient's condition. A typical threshold for diagnosing fatty liver disease as "alcohol-free" is a daily intake of less than 20g for women and less than 30g for men.

[0124] Specific diseases or health conditions associated with NAFLD include metabolic conditions such as diabetes, hypertension, obesity, dyslipidemia, dyslipidemia, glycogen storage disorders, Weber-Christian disease, acute fatty liver of pregnancy, and lipodystrophy. Other non-alcohol-related factors associated with fatty liver disease include malnutrition, complete parenteral nutrition, severe weight loss, refeeding syndrome, jejunal oil film bypass, gastric bypass, polycystic ovary syndrome, and diverticulosis.

[0125] Non-alcoholic steatohepatitis (NASH) is the most aggressive form of NAFLD, a condition characterized by excessive fat accumulation (steatosis) accompanied by inflammation of the liver. If left untreated, NASH can lead to the development of scar tissue in the liver (fibrosis) and ultimately cirrhosis. As described above, the compounds of the present invention have been found to be useful in the treatment of NAFLD and inflammation. Therefore, the compounds of the present invention are also useful in the treatment of NASH. Thus, in a further embodiment, the treatment is for non-alcoholic steatohepatitis (NASH).

[0126] AMPK activating compounds (e.g., 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide (i.e., the compound of formula II)) have been shown to be able to treat pain (Das V, et al. Reg Anesth Pain Med 2019;0:1-5. doi:10.1136 / rapm-2019-100839, and Das V, et al. Reg Anesth Pain Med 2019;0:1-6. doi:10.1136 / rapm-2019-100651), and these compounds can be considered analgesics. Therefore, since the compounds of the present invention can activate AMPK or are metabolized in vivo to form known AMPK activating compounds, the compounds of the present invention may be useful for treating pain. In particular, the compounds of the present invention may be useful in treating patients with severe pain or chronic pain, or in managing postoperative pain.

[0127] Opioid-based therapies such as opioid analgesics are used to treat severe chronic cancer pain, acute pain (e.g., recovery from surgery and breakthrough pain), and their use is increasing in the management of chronic non-malignant pain. However, the increased use of opioid-based therapies to treat pain has led to a rise in opioid dependence (e.g., opioid addiction). As AMPK activators, the compounds of the present invention can be used to treat pain as an alternative to opioid-based therapies, as is known to those skilled in the art. Accordingly, the compounds of the present invention may be useful for treating opioid addiction.

[0128] Specific autoimmune diseases known to those skilled in the art include Crohn's disease / inflammatory bowel disease, systemic lupus erythematosus, and type 1 diabetes.

[0129] Specific intestinal diseases that should be mentioned include Crohn's disease / inflammatory bowel disease and cancers of the gastrointestinal tract.

[0130] Those skilled in the art will understand that references to "treatment" (or similarly, "treating" a health condition) of a particular health condition take their ordinary meaning in the medical field. In particular, the term may refer to achieving a reduction in the severity and / or frequency of occurrence of one or more clinical symptoms associated with a health condition, as determined by the attending physician supervising a subject having or susceptible to such symptoms.

[0131] Those skilled in the art will understand that the above treatment or prevention is performed in a subject in need thereof. The need of a subject for the above treatment or prevention may be assessed by those skilled in the art using routine techniques. In the context of the present invention, a "subject in need of" a compound of the present invention includes a subject suffering from a disorder or health condition that is ameliorated by activation of AMPK. As used herein, the terms "disease" and "disorder" (as well as similarly, the terms "health condition", "illness", "medical problem", etc.) may be used interchangeably.

[0132] While not wishing to be bound by theory, administration of the compounds of the present invention is believed to improve the bioavailability of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide in the systemic circulation. Administration of a formulation comprising the compound of the present invention has been shown to result in an approximately 10-fold increase in the plasma concentration of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide under certain circumstances, as compared to administration of a formulation containing 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazol-5-yl]benzamide.

[0133] The compounds of the present invention (and formulations thereof), whether for use in the indications noted above or otherwise, may have the advantages of being more effective, less toxic, having a longer duration of action, being more potent, having fewer side effects, being more readily absorbed, and / or having a better pharmacokinetic profile (e.g., higher oral bioavailability and / or lower clearance) and / or possessing other useful pharmacological, physical, or chemical properties over other therapies known in the prior art. In particular, the compounds of the present invention may have the advantage of being more effective in vivo and / or exhibiting favorable properties.

[0134] [Figure] The following drawings are provided to illustrate various aspects of the concepts of the present invention and are not intended to limit the scope of the present invention unless specified herein.

[0135] Figure 1 is a Western blot image showing that Compound 3 increases AMPK phosphorylation in a dose-dependent manner.

[0136] Figure 2 shows the comparative results of oral pharmacokinetic studies using Compound 1 and Compound 3.

[0137] Figure 3 shows the comparative results of oral pharmacokinetic studies using compounds 5, 6, 7, 9, 10, 13, 14, 16, and 17.

[0138] [Examples] The present invention will be described in more detail by the following non-limiting embodiments.

[0139] The reaction scheme described below is intended to provide a general description of the procedure employed in the preparation of the compounds of the present invention. The examples provided herein are provided to illustrate, but are not limited to, the preparation of the compounds of the present invention and the intermediates thereof.

[0140] All starting materials, building blocks, reagents, acids, bases, dehydrating agents, solvents, and catalysts used to synthesize the compounds of the present invention are commercially available or can be routinely prepared by procedures described in the literature, e.g., Houben-Weyl "Science of Synthesis" volumes 1-48, Georg Thieme Verlag and subsequent editions.

[0141] The reaction may be carried out in the presence of a suitable solvent or diluent or mixture thereof, in a manner known to those skilled in the art of organic synthesis. The reaction may also be carried out in the presence of an acid or base, as needed, with cooling or heating, in a temperature range, for example, from about -30°C to about 150°C. In some embodiments, the reaction is carried out in an open or closed reaction vessel and / or in an atmosphere of an inert gas, such as nitrogen, in a temperature range of about 0°C to about 100°C, more specifically from room temperature to about 80°C.

[0142] [Abbreviation] The abbreviations used herein will be known to those skilled in the art. In particular, the following abbreviations may be used herein: AUC: Area under the concentration-time curve aq: water-based bw: weight C max : Peak plasma concentration d: doublet DCC:N,N'-Dicyclohexylcarbodiimide DCM:Dichloromethane DMAP:4-Dimethylaminopyridine DMSO:Dimethyl sulfoxide eq:Equivalent ESI:Electrospray ionization Et3N:Triethylamine EtOH:Ethanol g:Gram h:Hour HPLC:High-performance liquid chromatography LC:Liquid chromatography LCMS:Liquid chromatography-mass spectrometry LC-MS / MS:Liquid chromatography-(tandem) mass spectrometry LLOQ:Lower limit of quantitation m:Multiplet MeOH:Methanol MeOD:Methanol-d4 Min:Minute mL:Milliliter MRT:Mean residence time ND:Not detected NMR:Nuclear magnetic resonance RT:Room temperature s:Singlet T 1 / 2 :Half-life TBDMS-Cl:tert-Butyldimethylsilyl chloride T max :Time to reach peak plasma concentration THF:Tetrahydrofuran

[0143]

Table 1

[0144] The present invention is further described with reference to the following examples, which are not intended to limit the scope of the present invention.

[0145] [Example 1] Preparation of dimethylaminoacetic acid 5-[(Z)-4-chloro-benzoylimino]-2-(4-chloro-benzyl)-3-oxo-[1,2,4]thiadiazolidine-4-ylmethyl ester (compound 3) [ka]

[0146] Compound 1: 4-Chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide Compound 1 was prepared according to the procedure described in WO2011 / 004162.

[0147] Compound 2: 4-Chloro-N-{2-[(4-chlorophenyl)methyl]-4-(hydroxymethyl)-3-oxo-1,2,4-thiadiazolidine-5-ylidene}benzamide To a stirred solution of compound 1 (100 g, 0.26 mol) in THF (2.0 L), Et3N (150 mL, 1.05 mol) was added, followed by 37% formaldehyde (85 mL, 1.05 mol). The reaction mixture was stirred in RT for 12-14 hours.

[0148] Next, the reaction mixture was concentrated to remove THF. The resulting starting material was treated with toluene (2 × 100 mL) and dried under reduced pressure at 35-40°C to obtain compound 2 as a white solid (103 g, yield 95.5%).

[0149] Compound 3: Dimethylaminoacetic acid 5-[(Z)-4-chloro-benzoylimino]-2-(4-chloro-benzyl)-3-oxo-[1,2,4]thiadiazolidine-4-ylmethyl ester To a stirred solution of N,N-dimethylglycine (50.3 g, 0.48 mol) in THF (2.0 L), DMAP (8.93 g, 0.07 mol) and DCC (100.6 g, 0.48 mol) were added. After 10 minutes, compound 2 (200 g, 0.49 mol) was added to the reaction mixture in four portions over 30 minutes. The reaction mixture was stirred at 40-50°C for 16-18 hours. The reaction mixture was cooled to 20-30°C, filtered with Celite®, washed with THF (200 mL), and the filtrate was concentrated to remove THF. The obtained starting material was slurryed using MeOH (300 mL) at RT for 30 minutes, filtered, and washed with MeOH (50 mL). The recovered solid was dried under reduced pressure at 35-40°C to obtain compound 3 as a white solid (102 g, yield 42%). 1 H NMR (400 MHz, DMSO) δ (ppm): 2.20 (s, 6H), 3.21 (s, 2H), 4.84 (s, 2H), 6.03 (s, 2H), 7.40 (dd, 4H), 7.63 (d, 2H), 8.15 (d, 2H) LCMS: 495.2 [M+H] HPLC: 96.3% purity at 8.57 minutes.

[0150] [Example 2] Preparation of citrate (compound 4) of dimethylaminoacetic acid 5-[(Z)-4-chloro-benzoylimino]-2-(4-chloro-benzyl)-3-oxo-[1,2,4]thiadiazolidine-4-ylmethyl ester [ka]

[0151] To a stirred solution of dimethylaminoacetic acid 5-[(Z)-4-chloro-benzoylimino]-2-(4-chloro-benzyl)-3-oxo-[1,2,4]thiadiazolidine-4-ylmethyl ester (compound 3; 550 mg, 1.1 mmol) in acetone (30.0 v), 0.9 eq. citric acid was added. The reaction was stirred at RT for 2.3 hours. Next, the precipitated compound was collected by filtration and dried to obtain compound 4 (450 mg). 1 H NMR (400 MHz, DMSO) δ (ppm): 2.20 (s, 6H), 3.21 (s, 2H), 4.84 (s, 2H), 6.03 (s, 2H), 7.40 (dd, 4H), 7.63 (d, 2H), 8.15 (d, 2H) LCMS: 495.2 [M+H] HPLC: 98.3% purity at 9.95 minutes.

[0152] [Example 3] Preparation of 2-({[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methoxy}carbonyl)pyrrolidine-1-ium trifluoroacetate (Compound 5) [ka]

[0153] (Step 1) 1-[(tert-butoxy)carbonyl]pyrrolidine-2-carboxylic acid (compound A2) To a suspension of L-proline (A1) (1.0 g, 8.68 mmol) in DCM (10 mL), triethylamine (1.34 mL, 9.55 mmol) was added at 0°C, followed by di-tert-butyl dicarbonate (2.18 mL, 9.55 mmol). The resulting reaction mixture was stirred at RT for 16 hours. The reaction mixture was carefully acidified to pH 3 with a 5% citric acid solution. The product was extracted with ethyl acetate, the extract was dried, and evaporated under vacuum to obtain compound A2 as a white solid (1.55 g, yield 83%).

[0154] (Step 2) 1-tert-butyl 2-[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidin-4-yl]methylpyrrolidine-1,2-diculfonxylate (Compound A3) To a stirred solution of compound A2 (0.55 g, 2.56 mmol) in THF (11 mL, 20 V), DMAP (0.06 g, 0.49 mmol) and then DCC (0.52 g, 2.56 mmol) were added at 0-5°C and the mixture was stirred for 20 minutes. Compound 2 (0.7 g, 1.7 mmol) was added to the reaction mixture at 0-5°C, and the reaction mixture was stirred at ambient temperature (25-30°C) for 16 hours. The reaction mixture was filtered, the filtrate was concentrated, and the filtrate was purified by silica gel column chromatography to obtain compound A3 as a white solid (0.5 g, yield 50%).

[0155] (Step 3) 2-({[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methoxy}carbonyl)pyrrolidine-1-ium trifluoroacetate (Compound 5) To a stirred solution of compound A3 (0.5 g, 0.82 mmol) in dichloromethane (5 mL), trifluoroacetic acid (1 mL, 2.0 vol) was added at 0-5°C. The reaction mixture was stirred at ambient temperature (25-30°C) for 16 hours. The reaction mixture was concentrated to remove the dichloromethane. The resulting residue was reprecipitation with methanol / ether and filtered under vacuum to obtain compound 5 as a white solid (0.2 g, yield 39%). 1 H NMR (400 MHz, DMSO) δ (ppm): 1.90 (m, 2H), 1,98 (m, 1H), 2.22 (m, 1H), 3,18 (m, 2H), 4.48 (m, 1H), 4.84 (s, 2H), 6.13 (s, 2H), 7.42 (dd, 4H), 7.63 (d, 2H), 8.16 (d, 2H) LCMS: 507.3 [M+H] HPLC: Purity 95.3% at 10.07 minutes.

[0156] In step 1 of this embodiment, L-proline, i.e., the L-enantiomer of a naturally occurring amino acid, was used. In all subsequent embodiments, the L-enantiomer of each naturally occurring amino acid (or its alkylated and / or protected derivatives) was used.

[0157] [Example 4] Preparation of 6-{[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methoxy}-6-oxohexane-1,5-bis(aminium)ditrifluoroacetate (compound 6) [ka]

[0158] (Step 1) [5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methyl 2,6-bis({[(tert-butoxy)carbonyl]amino})hexanoate (compound B2) To a stirred solution of 2,6-bis({[(tert-butoxy)carbonyl]amino})hexanoic acid (compound B1; 0.84 g, 2.43 mmol) in THF (17 mL, 20 V), DMAP (0.074 g, 0.60 mmol) and then DCC (0.50 g, 2.43 mmol) were added at 0-5°C and the mixture was stirred for 20 minutes. Compound 2 (0.5 g, 1.21 mmol) was added to the reaction mixture at 0-5°C and the reaction mixture was stirred at ambient temperature (25-30°C) for 16 hours. The reaction mixture was filtered, the filtrate was concentrated, and the filtrate was purified by silica gel column chromatography to obtain compound B2 as a white solid (0.5 g, yield 55%).

[0159] (Step 2) 6-{[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methoxy}-6-oxohexane-1,5-bis(aminium)di-trifluoroacetate(6) To a stirred solution of compound B2 (0.5 g, 0.67 mmol) in dichloromethane (5 mL), trifluoroacetic acid (1 mL, 2.0 vol) was added at 0-5°C. The reaction mixture was stirred at ambient temperature (25-30°C) for 16 hours. The reaction mixture was concentrated to remove the dichloromethane. The resulting residue was reprecipitation with acetone / hexane and filtered under vacuum to obtain compound 6 as a white solid (0.28 g, yield 63%). 1 H NMR (400 MHz, DMSO) δ (ppm): 1.40 (m, 4H), 1,75 (m, 2H), 2.62 (m, 2H), 4,05 (m, 1H), 4.85 (s, 2H), 6.13 (s, 2H), 7.42 (dd, 4H), 7.63 (d, 2H), 8.16 (d, 2H) LCMS: 538.3 [M+H] HPLC: 95.0% purity at 8.28 minutes.

[0160] [Example 5] Preparation of (6-azaniumyl)-1-{[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methoxy}-1-oxohexane-2-yl)dimethylazanium ditrifluoroacetate (Compound 7) [ka]

[0161] (Step 1) 6-{[(tert-butoxy)carbonyl]amino}-2-(dimethylamino)hexanoic acid (compound C2) To a solution of Boc-Lys-OH(C1) (2.0 g, 8.1 mmol) in methanol (20 mL), 37% formaldehyde (2.6 mL, 32.4 mmol) was added, followed by Pd / C (0.2 g, 10% w / w). The resulting mixture was hydrogenated for 16 hours using a parr apparatus (5 kg, H2 pressure). After the reaction was complete, the reaction mixture was filtered through Celite® and concentrated under reduced pressure. The residue was powdered with ether to obtain compound C2 as a white solid (1.5 g, yield 67%).

[0162] (Step 2) [5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methyl 6-{[(tert-butoxy)carbonyl]amino}-2-(dimethylamino)hexanoate (compound C3) To a stirred solution of compound C2 (0.66 g, 2.43 mmol) in THF (14 mL, 20 V), DMAP (0.035 g, 0.29 mmol) and then DCC (0.50 g, 2.43 mmol) were added at 0-5°C and the mixture was stirred for 20 minutes. Compound 2 (0.4 g, 0.97 mmol) was added to the reaction mixture at 0-5°C, and the reaction mixture was stirred at ambient temperature (25-30°C) for 16 hours. The reaction mixture was filtered, the filtrate was concentrated, and the filtrate was purified by reverse-phase column chromatography (8:2 acetonitrile and water) to obtain compound C3 as a white solid (0.18 g, yield 64.9%).

[0163] (Step 3) (6-Azaniumyl-1-{[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methoxy}-1-oxohexane-2-yl)dimethylazanium di-trifluoroacetate (Compound 7) To a stirred solution of compound C3 (0.18 g, 0.027 mmol) in dichloromethane (3.6 mL), trifluoroacetic acid (0.36 mL, 2.0 vol) was added at 0-5°C. The reaction mixture was stirred at ambient temperature (25-30°C) for 3 hours. The reaction mixture was concentrated to remove the dichloromethane. The resulting residue was reprecipitation with acetone / hexane and filtered under vacuum to obtain compound 7 as an off-white solid (0.15 g, yield 81%). 1 H NMR (400 MHz, D2O) δ (ppm): 1.24 (m, 2H), 1,39 (m, 2H), 1.80 (m, 2H), 2.73 (s, 6H), 4.77 (s, 2H), 6.16 (m, 2H), 7.35 (dd, 4H), 7.58 (d, 2H), 8.12 (d, 2H) LCMS: 566.4 [M+H] HPLC: 97.5% purity at 8.09 minutes.

[0164] [Example 6] Preparation of (2-{[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methoxy}2-oxoethyl)(methyl)azanium trifluoroacetate (Compound 8) [ka]

[0165] (Step 1) [5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methyl 2-{[(tert-butoxy)carbonyl](methyl)amino}acetate (D2) To a stirred solution of N-Boc sarcosine (compound D1; 0.57 g, 3.04 mmol) in THF (12 mL, 20 V), DMAP (0.044 g, 0.36 mmol) and then DCC (0.62 g, 3.04 mmol) were added at 0-5°C and the mixture was stirred for 20 minutes. Compound 2 (0.5 g, 1.21 mmol) was added to the reaction mixture at 0-5°C, and the reaction mixture was stirred at ambient temperature (25-30°C) for 16 hours. The reaction mixture was filtered, the filtrate was concentrated, and the filtrate was purified by reverse-phase column chromatography to obtain compound D2 as a white solid (0.22 g, yield 31%).

[0166] (Step 2) (2-{[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methoxy}-2-oxoethyl)(methyl)azanium trifluoroacetate (Compound 8) To a stirred solution of compound D2 (0.2 g, 0.34 mmol) in dichloromethane (4.0 mL), trifluoroacetic acid (0.4 mL, 2.0 vol) was added at 0-5°C. The reaction mixture was stirred at ambient temperature (25-30°C) for 16 hours. The reaction mixture was concentrated to remove the dichloromethane. The resulting residue was powdered with diethyl ether to obtain compound 8 as a white solid (0.17 g, yield 84%). 1 H NMR (400 MHz, DMSO) δ (ppm): 2.57 (s, 3H), 4.09 (s, 2H), 4.85 (s, 2H), 6.13 (m, 2H), 7.42 (dd, 4H), 7.64 (d, 2H), 8.17 (d, 2H) LCMS: 481.3 [M+H] HPLC: 98.6% purity at 8.91 minutes.

[0167] [Example 7] Preparation of 2-{[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methoxy}2-oxoethane-1-aminium trifluoroacetate (Compound 9) [ka]

[0168] (Step 1) [5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methyl 2-{[(tert-butoxy)carbonyl]amino}acetate (compound E2) To a stirred solution of N-Boc glycine (compound E1; 0.53 g, 3.04 mmol) in THF (11 mL, 20 V), DMAP (0.044 g, 0.36 mmol) and then DCC (0.62 g, 3.04 mmol) were added at 0-5°C and the mixture was stirred for 20 minutes. Compound 2 (0.5 g, 1.21 mmol) was added to the reaction mixture at 0-5°C and the reaction mixture was stirred at ambient temperature (25-30°C) for 16 hours. The reaction mixture was filtered, the filtrate was concentrated, and the filtrate was purified by reverse-phase column chromatography to obtain compound E2 as a white solid (0.32 g, yield 46%).

[0169] (Step 2) 2-{[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidin-4-yl]methoxy}2-oxoethane-1-aminium trifluoroacetate (Compound 9) To a stirred solution of compound E2 (0.3 g, 0.52 mmol) in dichloromethane (3.0 mL), trifluoroacetic acid (0.6 mL, 2.0 vol) was added at 0-5°C. The reaction mixture was stirred at ambient temperature (25-30°C) for 2 hours. The reaction mixture was concentrated to remove the dichloromethane. The resulting residue was powdered with diethyl ether to obtain compound 9 as a white solid (0.23 g, yield 76%). 1 H NMR (400 MHz, DMSO) δ (ppm): 3.91 (s, 2H), 4.85 (s, 2H), 6.13 (m, 2H), 7.42 (dd, 4H), 7.64 (d, 2H), 8.17 (d, 2H) LCMS: 467.2 [M+H] HPLC: 98.2% purity at 8.55 minutes.

[0170] [Example 8] Preparation of 2-({[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methoxy}carbonyl)-1-methylpyrrolidine (Compound 10) [ka]

[0171] (Step 1) 1-Methylpyrrolidine-2-carboxylic acid (Compound F1) To a solution of L-proline (compound A1; 2.0 g, 17.37 mmol) in methanol (20 mL), 37% formaldehyde (1.54 mL, 19.1 mmol) was added, followed by Pd / C (0.5 g, 25% w / w). The resulting mixture was hydrogenated in a Parr apparatus (1 kg H2 pressure) for 16 hours. After the reaction was complete, the reaction mixture was filtered through Celite® and concentrated under reduced pressure. The residue was powdered with ether to obtain compound F1 as a white solid (1.9 g, yield 84%).

[0172] (Step 2) 2-({[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methoxy}carbonyl)-1-methylpyrrolidine-1-ium trifluoroacetate (Compound 10) To a stirred solution of compound F1 (0.39 g, 3.0 mmol) in THF (10 mL, 20 V), DMAP (0.074 g, 0.60 mmol) and then DCC (0.62 g, 3.04 mmol) were added at 0-5°C and the mixture was stirred for 20 minutes. Compound 2 (0.5 g, 0.1.21 mmol) was added to the reaction mixture at 0-5°C and the reaction mixture was stirred at ambient temperature (25-30°C) for 16 hours. The reaction mixture was filtered, the filtrate was concentrated and purified by reverse-phase column chromatography, and then reprecipitation using acetone / diethyl ether to obtain compound 10 as a white solid (0.25 g, yield 39%). 1H NMR (400 MHz, DMSO) δ (ppm): 1.83 (br, 1H), 2,03 (m, 2h), 2.40 (br, 1H), 2,87 (br, 1H), 3,13 (br, 1H), 3.57 (br, 1H), 4.85 (s, 2H), 6.13 (m, 2H), 7.41 (dd, 4H), 7.64 (d, 2H), 8.15(d, 2H) LCMS: 521.3 [M+H] HPLC: 96.9% purity at 9.16 minutes.

[0173] [Example 9] Preparation of 1-{[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methoxy}-1-oxopropane-2-aminium trifluoroacetate (Compound 11) [ka]

[0174] (Step 1) [5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methyl 2-{[(tert-butoxy)carbonyl]amino}propanoate (compound G2) To a stirred solution of N-Boc alanine (compound G1) (0.57 g, 3.04 mmol) in THF (10 mL, 20 V), DMAP (0.044 g, 0.36 mmol) and then DCC (0.62 g, 3.04 mmol) were added at 0-5°C and the mixture was stirred for 20 minutes. Compound 2 (0.5 g, 1.21 mmol) was added to the reaction mixture at 0-5°C, and the reaction mixture was stirred at ambient temperature (25-30°C) for 16 hours. The reaction mixture was filtered, the filtrate was concentrated, and the filtrate was purified by reverse-phase column chromatography to obtain compound G2 as a white solid (0.31 g, yield 43%).

[0175] (Step 2) 1-{[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidin-4-yl]methoxy}-1-oxopropane-2-aminium trifluoroacetate (Compound 11) To a stirred solution of compound G2 (0.3 g, 0.51 mmol) in dichloromethane (3.0 mL), trifluoroacetic acid (0.9 mL, 3.0 vol) was added at 0-5°C. The reaction mixture was stirred at ambient temperature (25-30°C) for 2 hours. The reaction mixture was concentrated to remove the dichloromethane. The resulting residue was powdered with diethyl ether to obtain compound 11 as a white solid (0.14 g, yield 45%). 1 H NMR (400 MHz, DMSO) δ (ppm): 1.33 (d, 3H), 4.19 (t, 1h), 4.85 (s, 2H), 6.12 (m, 2H), 7.41 (dd, 4H), 7.64 (m, 1H), 8.15(d, 2H) LCMS: 481.3 [M+H] HPLC: Purity 95.7% at 9.14 minutes.

[0176] [Example 10] Preparation of 1-{[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methoxy}-3-methyl-1-oxopentane-2-aminium trifluoroacetate (Compound 12) [ka]

[0177] (Step 1) [5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methyl 2-{[(tert-butoxy)carbonyl]amino}-3-methylpentanoate (compound H2) To a stirred solution of N-Boc isoleucine (compound H1; 0.56 g, 3.04 mmol) in THF (10 mL, 20 V), DMAP (0.044 g, 0.36 mmol) and then DCC (0.62 g, 3.04 mmol) were added at 0-5°C and the mixture was stirred for 20 minutes. Compound 2 (0.5 g, 1.21 mmol) was added to the reaction mixture at 0-5°C, and the reaction mixture was stirred at ambient temperature (25-30°C) for 16 hours. The reaction mixture was filtered, the filtrate was concentrated, and the filtrate was purified by reverse-phase column chromatography to obtain compound H2 as a white solid (0.4 g, yield 52%).

[0178] (Step 2) 1-{[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidin-4-yl]methoxy}-3-methyl-1-oxopentane-2-aminium trifluoroacetate (Compound 12) To a stirred solution of compound H2 (0.4 g, 0.64 mmol) in dichloromethane (3.0 mL), trifluoroacetic acid (1.2 mL, 3.0 vol) was added at 0-5°C. The reaction mixture was stirred at ambient temperature (25-30°C) for 2 hours. The reaction mixture was concentrated to remove the dichloromethane. The resulting residue was powdered with diethyl ether to obtain compound 12 as a white solid (0.22 g, yield 53%). 1 H NMR (400 MHz, DMSO) δ (ppm): 0.67 (t, 3H), 0.84 (d, 2H), 1.15 (m, 1H), 1.33 (m, 1H), 1.81 (br, 1H), 4.10 (s, 1h), 4.86 (s, 2H), 6.03 (d, 1H), 6.20 (d, 1H), 7.45 (dd, 4H), 7.64 (d, 1H), 8.17(d, 2H) LCMS: 523.3 [M+H] HPLC: 98.0% purity at 10.38 minutes.

[0179] [Example 11] Preparation of [5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methyl 2-(dimethylamino)-3-phenylpropanoate (compound 13) [ka]

[0180] (Step 1) 2-(dimethylamino)-3-phenylpropanoic acid (compound I2) To a solution of Phe-Ala-OH (compound I1; 1.0 g, 6.05 mmol) in methanol (20 mL), 37% formaldehyde (1.96 mL, 24.21 mmol) was added, followed by Pd / C (0.1 g, 10% w / w). The resulting mixture was hydrogenated for 16 hours using a parr apparatus (5 kg H2 pressure). After the reaction was complete, the reaction mixture was filtered through Celite® and concentrated under reduced pressure. The residue was powdered with ether to obtain compound I2 as a white solid (1.0 g, yield 86%).

[0181] (Step 2) [5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidin-4-yl]methyl 2-(dimethylamino)-3-phenylpropanoate (Compound 13) To a stirred solution of compound I2 (0.47 g, 2.43 mmol) in THF (10 mL, 20 V), DMAP (0.044 g, 0.36 mmol) and then DCC (0.62 g, 3.04 mmol) were added at 0-5°C and the mixture was stirred for 20 minutes. Compound 2 (0.5 g, 1.21 mmol) was added to the reaction mixture at 0-5°C and the reaction mixture was stirred at ambient temperature (25-30°C) for 16 hours. The reaction mixture was filtered, the filtrate was concentrated, and the filtrate was purified by reverse-phase column chromatography to obtain compound 13 as a white solid (0.22 g, yield 30%). 1H NMR (400 MHz, DMSO) δ (ppm): 2.49 (s, 6H), 2.92 (m, 2h), 3.51 (t, 1H), 4.89 (s, 2h), 5.97 (m, 2H), 7.07 (m, 5H), 7.40 (d, 2H), 7.45 (d, 2H), 7.64 (d, 2H), 8.15 (d, 2H) LCMS: 586.0 [M+H] HPLC: 97.5% purity at 11.54 minutes.

[0182] [Example 12] Preparation of 1-{[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methoxy}-1-oxo-3-phenylpropane-2-aminium trifluoroacetate (Compound 14) [ka]

[0183] (Step 1) 2-{[(tert-butoxy)carbonyl]amino}-3-phenylpropanoic acid (compound J1) To a solution of Phe-Ala-OH (compound I1; 1.0 g, 6.05 mmol) in DCM (10 mL), triethylamine (1.70 mL, 12.17 mmol) was added at 0°C, followed by di-tert-butyl pyrocarbonate (2.18 mL, 9.08 mmol). The resulting reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was carefully acidified to pH 4 with a 5% citric acid solution. The product was extracted with DCM, the extract was dried, evaporated under vacuum, and purified by reverse-phase column chromatography to obtain compound J1 as a colorless wax (1.3 g, yield 81%).

[0184] (Step 2) [5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methyl 2-{[(tert-butoxy)carbonyl]amino}-3-phenylpropanoate (compound J2) To a stirred solution of compound J1 (0.64 g, 2.4 mmol) in THF (10 mL, 20 V), DMAP (0.044 g, 0.36 mmol) and then DCC (0.62 g, 3.04 mmol) were added at 0-5°C and the mixture was stirred for 20 minutes. Compound 2 (0.5 g, 1.21 mmol) was added to the reaction mixture at 0-5°C and the reaction mixture was stirred at ambient temperature (25-30°C) for 16 hours. The reaction mixture was filtered, the filtrate was concentrated, and the filtrate was purified by reverse-phase column chromatography to obtain compound J2 as a white solid (0.3 g, yield 37%).

[0185] (Step 3) 1-{[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidin-4-yl]methoxy}-1-oxo-3-phenylpropane-2-aminium trifluoroacetate (Compound 14) To a stirred solution of compound J2 (0.3 g, 0.45 mmol) in dichloromethane (3.0 mL), trifluoroacetic acid (0.9 mL, 3.0 vol) was added at 0-5°C. The reaction mixture was stirred at ambient temperature (25-30°C) for 2 hours. The reaction mixture was concentrated to remove the dichloromethane. The resulting residue was powdered with diethyl ether to obtain compound 14 as a white solid (0.2 g, yield 66%). 1 H NMR (400 MHz, DMSO) δ (ppm): 2.98 (m, 1H), 3.14 (m, 1H), 4.44 (m, 1H), 4.85 (s, 2h), 5.877 (s, 1H), 6.20 (d, 1H), 7.07 (m, 5H), 7.41 (d, 2H), 7.47 (d, 2H), 7.64 (d, 2H), 8.16 (d, 2H) LCMS: 558.0 [M+H] HPLC: 98.6% purity at 10.47 minutes.

[0186] [Example 13] Preparation of 1-{[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methoxy}-4-methyl-1-oxopentane-2-aminium trifluoroacetate (Compound 15) [ka]

[0187] (Step 1) [5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methyl 2-{[(tert-butoxy)carbonyl]amino}-4-methylpentanoate (compound K2) To a stirred solution of 2-{[(tert-butoxy)carbonyl]amino}-4-methylpentanoic acid (compound K1; 0.56 g, 2.43 mmol) in THF (10 mL, 20 V), DMAP (0.044 g, 0.36 mmol) and then DCC (0.62 g, 3.04 mmol) were added at 0-5°C and the mixture was stirred for 20 minutes. Compound 2 (0.5 g, 1.21 mmol) was added to the reaction mixture at 0-5°C and the reaction mixture was stirred at ambient temperature (25-30°C) for 16 hours. The reaction mixture was filtered, the filtrate was concentrated, and the filtrate was purified by reverse-phase column chromatography to obtain compound K2 as a white solid (0.4 g, yield 75%).

[0188] (Step 2) 1-{[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methoxy}-4-methyl-1-oxopentane-2-aminium trifluoroacetate (Compound 15) To a stirred solution of compound K2 (0.35 g, 0.56 mmol) in dichloromethane (3.0 mL), trifluoroacetic acid (1.05 mL, 3.0 vol) was added at 0-5°C. The reaction mixture was stirred at ambient temperature (25-30°C) for 2 hours. The reaction mixture was concentrated to remove the dichloromethane. The resulting residue was powdered with diethyl ether to obtain compound 15 as a white solid (0.27 g, yield 93%). 1 H NMR (400 MHz, DMSO) δ (ppm): 2.98 (m, 1H), 3.14 (m, 1H), 4.44 (m, 1H), 4.85 (s, 2h), 5.877 (s, 1H), 6.20 (d, 1H), 7.07 (m, 5H), 7.41 (d, 2H), 7.47 (d, 2H), 7.64 (d, 2H), 8.16 (d, 2H) LCMS: 523.4 [M+H] HPLC: 98.0% purity at 10.56 minutes.

[0189] [Example 14] Preparation of 1-{[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methoxy}3-hydroxy-1-oxopropane-2-aminium trifluoroacetate (Compound 16) [ka]

[0190] (Step 1) 2-{[(tert-butoxy)carbonyl]amino}-3-[(tert-butyldimethylsilyl)oxy]propanoic acid (compound L2) To a solution of Boc-Ser-OH (compound L1; 1.0 g, 4.87 mmol) in DCM (10 mL), imidazole (0.53 mg, 7.79 mmol) was added at 0°C, followed by TBDMS-Cl (1.10 g, 7.30 mmol). The resulting reaction mixture was stirred at room temperature for 16 hours. The product was extracted with DCM, the extract was dried, evaporated under vacuum, and purified by reverse-phase column chromatography to obtain compound L2 as a colorless wax (0.7 g, 46% yield).

[0191] (Step 2) [5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methyl 2-{[(tert-butoxy)carbonyl]amino}-3-[(tert-butyldimethylsilyl)oxy]propanoate (compound L3) To a stirred solution of compound L2 (0.77 g, 2.43 mmol) in THF (10 mL, 20 V), DMAP (0.044 g, 0.36 mmol) and then DCC (0.62 g, 3.04 mmol) were added at 0-5°C and the mixture was stirred for 20 minutes. Compound 2 (0.5 g, 1.21 mmol) was added to the reaction mixture at 0-5°C, and the reaction mixture was stirred at ambient temperature (25-30°C) for 16 hours. The reaction mixture was filtered, the filtrate was concentrated, and the filtrate was purified by reverse-phase column chromatography to obtain compound L3 as a white solid (0.4 g, yield 46%).

[0192] (Step 3) 1-{[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methoxy}-3-hydroxy-1-oxopropane-2-aminium trifluoroacetate (Compound 16) To a stirred solution of compound L3 (0.4 g, 0.56 mmol) in dichloromethane (4.0 mL), trifluoroacetic acid (1.2 mL, 3.0 vol) was added at 0-5°C. The reaction mixture was stirred at ambient temperature (25-30°C) for 2 hours. The reaction mixture was concentrated to remove the dichloromethane. The resulting residue was purified by reverse-phase column chromatography to obtain compound 16 as a white solid (0.1 g, yield 29%). 1 H NMR (400 MHz, DMSO) δ (ppm): 3.80 (m, 2H), 4.27 (m, 1H), 4.85 (s, 2h), 5.60 (t, 1H), 6.12 (q, 2H), 7.07 (m, 5H), 7.44 (dd, 4H), 7.63 (d, 2H), 8.18 (d, 2H) LCMS: 497.3 [M+H] HPLC: 91.6% purity at 8.52 minutes.

[0193] [Example 15] Preparation of 3-carboxy-1-{[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methoxy}-1-oxopropane-2-aminium trifluoroacetate (Compound 17) [ka]

[0194] (Step 1) 4-(tert-butoxy)-2-{[(tert-butoxy)carbonyl]amino}-4-oxobutanoic acid (compound M2) To a solution of 4-tert-butyl L-aspartate (compound M1; 2.0 g, 10.5 mmol) in dioxane:water (7:3, 20 mL), NaOH (0.84 g, 21.1 mmol) was added, followed by di-tert-butyl pyrocarbonate (3.64 mL, 15.85 mmol). The resulting reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was carefully acidified to pH 3 with 1.5 N HCl solution. The product was extracted with ethyl acetate, and the extract was dried and evaporated under vacuum to obtain compound M2 as a white solid (2.5 g, 81% yield).

[0195] (Step 2) tert-butyl 1-[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methyl 2-{[(tert-butoxy)carbonyl]amino}butanedioate (compound M3) To a stirred solution of compound M2 (0.88 g, 3.04 mmol) in THF (18 mL, 20 V), DMAP (0.044 g, 0.36 mmol) and then DCC (0.62 g, 3.04 mmol) were added at 0-5°C and the mixture was stirred for 20 minutes. Compound 2 (0.5 g, 1.21 mmol) was added to the reaction mixture at 0-5°C and the reaction mixture was stirred at ambient temperature (25-30°C) for 16 hours. The reaction mixture was filtered, the filtrate was concentrated, and the filtrate was purified by reverse-phase column chromatography to obtain compound M3 as a white solid (0.45 g, yield 54%).

[0196] (Step 3) 3-Carboxy-1-{[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methoxy}-1-oxopropane-2-aminium trifluoroacetate (Compound 17) To a stirred solution of compound M3 (0.45 g, 0.66 mmol) in dichloromethane (4.5 mL), trifluoroacetic acid (0.9 mL, 2.0 vol) was added at 0-5°C. The reaction mixture was stirred at ambient temperature (25-30°C) for 6 hours. The reaction mixture was concentrated to remove the dichloromethane. The resulting residue was purified by reverse-phase column chromatography to obtain compound 17 as a white solid (0.12 g, yield 28%). 1 H NMR (400 MHz, DMSO) δ (ppm): 2.85 (m, 2H), 4.41 (m, 1H), 4.84 (s, 2h), 6.12 (dd, 2H), 6.12 (q, 2H), 7.45 (dd, 4H), 7.63 (d, 2H), 8.18 (d, 2H) LCMS: 525.3 [M+H] HPLC: Purity 94.2% at 8.29 minutes.

[0197] [Example 16] Preparation of [5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methyl 2-[(2S)-2-azaniumyl-3-phenylpropanamide]acetate chloride (compound 18) [ka]

[0198] (Step 2) Preparation of 2-[(2S)-2-{[(tert-butoxy)carbonyl]amino}-3-phenylpropanamide]methyl acetate (N2) To a stirred solution of (2S)-2-{[(tert-butoxy)carbonyl]amino}-3-phenylpropanoic acid (J1 synthesized as described above; 2.6 g, 9.8 mmol) in DCM (52 mL, 20 V), DCC (2.42 g, 11.76 mmol), DMAP (119 mg, 0.98 mmol), and triethylamine (2.71 mL, 19.6 mmol) were added at 0-5°C and the mixture was stirred for 20 minutes. 2-Methyl aminoacetate hydrochloride (compound N1) (1.6 g, 12.74 mmol) was added to the reaction mixture at 0-5°C and the reaction mixture was stirred at ambient temperature (25-30°C) for 16 hours. The reaction product was filtered to remove by-products (urea), the filtrate was concentrated, and the filtrate was purified by silica gel (60-120 mesh) column chromatography to obtain compound N2 as an off-white solid (2.73 g, yield 83%).

[0199] (Step 3) Preparation of 2-[(2S)-2-{[(tert-butoxy)carbonyl]amino}-3-phenylpropanamide]acetic acid (compound N3) To a stirred solution of compound N2 (3.0 g, 8.91 mmol) in methanol (30 mL, 10 V), 12 mL of 2 M NaOH solution (4 V) was added at 0-5°C, and the reaction mixture was stirred at ambient temperature (25-30°C) for 16 hours. The reaction product was concentrated to remove volatiles, diluted with water (50 mL), and washed with MTBE (100 mL). The aqueous layer was carefully acidified to pH 4-5 with 1.5 N HCl solution and extracted with ethyl acetate (200 mL x 2). The organic layers were combined, dried over Na2SO4, and concentrated under reduced pressure. The resulting residue was washed with hexane (50 mL x 2) to obtain compound N3 as a white solid (1.6 g, yield 56%).

[0200] (Step 4) Preparation of [5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methyl 2-[(2S)-2-{[(tert-butoxy)carbonyl]amino}-3-phenylpropanamide]acetate (compound N4) To a stirred solution of compound N3 (1.6 g, 4.96 mmol) in THF (32 mL, 20 V), DCC (1.53 g, 7.45 mmol) and then DMAP (0.3 g, 2.48 mmol) were added at 0-5°C and the mixture was stirred for 20 minutes. To the resulting reaction mixture, 4-chloro-N-{2-[(4-chlorophenyl)methyl]-4-(hydroxymethyl)-3-oxo-1,2,4-thiadiazolidine-5-ylidene}benzamide 6 (1.42 g, 3.47 mmol) was added at 0-5°C and the mixture was stirred at ambient temperature (25-30°C) for 16 hours. The reaction product was filtered, the filtrate was concentrated, and the mixture was purified by reverse-phase column chromatography to obtain compound N4 as a white solid (354 mg, yield 10%).

[0201] (Step 5) Preparation of [5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methyl 2-[(2S)-2-azaniumyl-3-phenylpropanamide]acetate chloride (compound 18) To a stirred solution of compound N4 (0.2 g, 0.279 mmol) in THF (1.0 mL, 5 vol), hydrochloric acid in diethyl ether (4.0 mL, 20 vol) was added at 0-5°C, and the reaction mixture was stirred at the same temperature for 20 minutes. The reaction mixture was concentrated to remove volatiles, and the resulting residue was powdered with diethyl ether to obtain compound 18 as a white solid (150 mg, yield 83%). 1 H NMR (300 MHz, DMSO) δ (ppm): 2.94 (m, 1H), 3.15 (m, 1H), 4.05 (m, 3h), 4.84 (s, 2H), 6.07 (s, 2H), 7.27 (m, 5H), 7.42 (q, 4H), 7.62 (d, 2H), 8.17 (m, 2H), 8,23 (br, 2h, NH), ), 9.14 (t, 1H, NH) LCMS: 614.31 [M+H] HPLC: 97.35% purity at 11.82 minutes.

[0202] [Example 17] Preparation of [5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methyl (2S)-2-[(2S)-2-azaniumyl-3-methylbutanamide]-3-methylbutanoate chloride (compound 19) [ka]

[0203] (Step 1) Preparation of [5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methyl (2S)-2-{[(tert-butoxy)carbonyl]amino}-3-methylbutanoate (compound O2) To a stirred solution of (2S)-2-{[(tert-butoxy)carbonyl]amino}-3-methylbutanoic acid (O1; 3.0 g, 13.8 mmol) in THF (60 mL, 20 V), DMAP (0.84 g, 6.91 mmol) and then DCC (4.27 g, 20.73 mmol) were added at 0-5°C and the mixture was stirred for 20 minutes. Compound 2 (2.83 g, 6.91 mmol) was added to the reaction mixture at 0-5°C, and the resulting reaction mixture was stirred at ambient temperature (25-30°C) for 16 hours. The reaction product was diluted with dichloromethane (200 mL), washed with water (2 × 100 mL), and concentrated under reduced pressure to obtain compound O2 as a white solid (6.0 g, 77%).

[0204] Note: The crude compound O2 was not purified before proceeding to the next step.

[0205] (Step 2) Preparation of (2S)-1-{[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methoxy}-3-methyl-1-oxobutan-2-aminium trifluoroacetate (compound O3) To a stirred solution of [5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methyl (2S)-2-{[(tert-butoxy)carbonyl]amino}-3-methylbutanoate (compound O2; 6.0 g, 9.85 mmol) in dichloromethane (25 mL), trifluoroacetic acid (24 mL, 4.0 vol) was added at 0-5°C, and the mixture was stirred at ambient temperature (25-30°C) for 4 hours. The reaction mixture was concentrated to remove volatiles, and the resulting residue was powdered with ether and filtered under vacuum to obtain compound O3 as a white solid (1.5 g, yield 24%).

[0206] (Step 3) Preparation of [5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methyl (2S)-2-(2-{[(tert-butoxy)carbonyl]amino}-3-methylbutanamide)-3-methylbutanoate (compound O4) To a stirred solution of (2S)-2-{[(tert-butoxy)carbonyl]amino}-3-methylbutanoic acid (compound O1; 1.30 g, 6.02 mmol) in THF (30 mL, 20 V), DMAP (0.088 g, 0.72 mmol) and then DCC (1.39 g, 6.75 mmol) were added at 0-5°C and stirred for 20 minutes. To the above reaction mixture, (2S)-1-{[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methoxy}-3-methyl-1-oxobutan-2-aminium trifluoroacetate (compound O3; 1.5 g, 2.41 mmol) was added at 0-5°C and stirred at ambient temperature (25-30°C) for 16 hours. The reaction mixture was filtered, the filtrate was concentrated, and the compound O4 was purified by reverse-phase column chromatography to obtain compound O4 as a white solid (0.65 g, yield 37%).

[0207] (Step 4) Preparation of [5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methyl (2S)-2-[(2S)-2-azaniumyl-3-methylbutanamide]-3-methylbutanoate trifluoroacetate (compound O5) To a stirred solution of [5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methyl (2S)-2-(2-{[(tert-butoxy)carbonyl]amino}-3-methylbutanamide)-3-methylbutanoate (compound O4; 0.66 g, 0.93 mmol) in dichloromethane (25 mL), trifluoroacetic acid (1.98 mL, 3.0 vol) was added at 0-5°C, and the reaction mixture was stirred at ambient temperature (25-30°C) for 4 hours. The reaction mixture was concentrated to remove volatile matter, and the resulting residue was powdered with ether and filtered under vacuum to obtain compound O5 as a white solid (0.45 g, yield 67%).

[0208] (Step 5) Preparation of [5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methyl (2S)-2-[(2S)-2-azaniumyl-3-methylbutanamide]-3-methylbutanoate chloride (compound 19) To a stirred solution of [5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methyl (2S)-2-[(2S)-2-azaniumyl-3-methylbutanamide]-3-methylbutanoate trifluoroacetate (compound O5; 0.45 g, 0.62 mmol) in THF (2.25 mL, 5.0 vol), HCl in diethyl ether (9.0 mL, 20 vol) was added at 0-5°C, and the reaction mixture was stirred at the same temperature for 20 minutes. The reaction product was concentrated to remove volatiles, and the resulting residue was powdered with diethyl ether to obtain compound 19 as a white solid (240 mg, yield 60%). 1H NMR (300 MHz, DMSO) δ (ppm): 0.8-0.9 (bm, 12H, -CH3), 2,07 (m, 2H), 3.7 (dd, 1H), 4.26 (m, 1H), 4.84 (s, 2h), 5.97 (dd, 1H), 6.11 (q, 1H), 7.43 (dd, 4H), 7.63 (d, 2H), 8.15 (m, 2K), 8.19 (m, 3H, NH3+), 8.6 (m, 1H, NH) LCMS: 608.53 [M+H] HPLC: Purity 79.5% at 11.125 min and 17.3% at 11.97 min (rotational isomers)

[0209] [Example 18] Preparation of [5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methyl (2R)-2-[(2R)-2-azaniumyl-3-methylbutanamide]-3-methylbutanoate chloride (compound 20) [ka]

[0210] The method used was similar to that described for the preparation of compound 19 in Example 17, and compound 20 was obtained as a white solid starting from (2R)-2-{[(tert-butoxy)carbonyl]amino}-3-methylbutanoic acid (300 mg, yield 88%). 1 H NMR (300 MHz, DMSO) δ (ppm): 0.8-0.9 (bm, 12H, -CH3), 2,07 (m, 2H), 3.69 (dd, 1H), 4.26 (m, 1H), 4.84 (s, 2h), 5.97 (dd, 1H), 6.11 (q, 1H), 7.43 (dd, 4H), 7.63 (d, 2H), 8.15 (m, 2K), 8.19 (m, 3H, NH3+), 8.6 (m, 1H, NH) LCMS: 608.53 [M+H] HPLC: Purity 75.93% at 13.43 min and 22.45% at 14.5 min (rotational isomers)

[0211] [Example 19] Preparation of [5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methyl 2-[(2S)-2,6-diazaniumylhexamide]acetate dichloride (compound 21) [ka]

[0212] (Step 1) Preparation of 2-[(2S)-2,6-bis({[(tert-butoxy)carbonyl]amino})hexaneamide]methyl acetate (compound P2) To a stirred solution of (2S)-2,6-bis({[(tert-butoxy)carbonyl]amino})hexanoic acid (compound B1; 2.6 g, 7.51 mmol) in DCM (52 mL, 20 V), DCC (3.86 g, 18.78 mmol), DMAP (275 mg, 2.25 mmol), and triethylamine (2.07 mL, 15.02 mmol) were added at 0-5°C and the mixture was stirred for 20 minutes. 2-aminomethyl hydrochloride P1 (939 mg, 7.51 mmol) was added to the reaction mixture at 0-5°C and the mixture was stirred at ambient temperature (25-30°C) for 16 hours. The reaction product was filtered to remove the by-product (urea), and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain compound P2 as a colorless liquid (1.81 g, yield 58%).

[0213] (Step 2) Preparation of 2-[(2S)-2,6-bis({[(tert-butoxy)carbonyl]amino})hexanamide]acetic acid (compound P3) To a stirred solution of 2-[(2S)-2,6-bis({[(tert-butoxy)carbonyl]amino})hexaneamide]methyl acetate (compound P2; 3.0 g, 7.19 mmol) in methanol (30 mL, 10 V), a 2 M NaOH solution (12 mL, 4 V) was added at 0-5°C. The resulting reaction mixture was stirred at ambient temperature (25-30°C) for 16 hours. The reaction product was concentrated to remove methanol, diluted with water (50 mL), and washed with MTBE (50 mL x 2). The aqueous layer was carefully acidified to pH 4-5 with 1.5 N aqueous HCl and extracted with ethyl acetate (100 mL x 2). The organic layers were combined, dried over Na2SO4, and concentrated under reduced pressure. The residue was washed with hexane to obtain compound P3 as a colorless liquid (2.75 g, yield 95%).

[0214] (Step 3) Preparation of [5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methyl 2-[(2S)-2,6-bis({[(tert-butoxy)carbonyl]amino})hexaneamide]acetate (compound P4) To a stirred solution of (2-[(2S)-2,6-bis({[(tert-butoxy)carbonyl]amino})hexaneamide]acetic acid (compound P3; 1.75 g, 4.34 mmol) in THF (35 mL, 20 V), DMAP (0.26 g, 2.17 mmol) and then EDC.HCl (1.24 g, 6.51 mmol) were added at 0-5°C and the mixture was stirred for 20 minutes. Compound 2 (0.89 g, 2.17 mmol) was added to the reaction mixture at 0-5°C and the mixture was stirred at ambient temperature (25-30°C) for 16 hours. The reaction product was diluted with dichloromethane (200 mL), washed with water (2 × 100 mL), and concentrated under reduced pressure. The residue obtained by evaporating the solvent was purified by reverse-phase column chromatography to obtain compound P4 as a white solid (0.3 g, yield 12%).

[0215] (Step 4) Preparation of [5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methyl 2-[(2S)-2,6-diazaniumylhexamide]acetate di-trifluoroacetate (compound P5) To a stirred solution of [5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methyl 2-[(2S)-2,6-bis({[(tert-butoxy)carbonyl]amino})hexaneamide]acetate (compound P4; 0.3 g, 0.37 mmol) in dichloromethane (4.5 mL, 15 V), trifluoroacetic acid (1.2 mL, 4 vol) was added at 0-5 °C, and the mixture was stirred at ambient temperature (25-30 °C) for 4 hours. The reaction mixture was concentrated to remove volatiles, the resulting residue was washed with ether, filtered, and dried under vacuum to obtain compound P5 as a white solid (0.25 g, yield 96%).

[0216] (Step 5) Preparation of [5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methyl 2-[(2S)-2,6-diazaniumylhexamide]acetate dichloride (compound 21) To a stirred solution of [5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methyl 2-[(2S)-2,6-diazaniumylhexanamide]acetate di-trifluoroacetate (compound P5; 0.11 g, 0.15 mmol) in THF (0.5 mL, 5.0 vol), HCl in diethyl ether (2.2 mL, 20 vol) was added at 0-5°C, and the reaction mixture was stirred at the same temperature for 20 minutes. The reaction product was concentrated to remove volatiles, and the resulting residue was powdered with diethyl ether to obtain compound 21 as a white solid (90 mg, yield 90%). 1H NMR (300 MHz, DMSO) δ (ppm): 1,44 (br, 2H), 1.60 (br, 2H), 1,75 (br, 2H), 2.78 (br, 2H), 3.87 (br, 1H), 4.04 (m, 2h), 4.86 (s, 2H), 6.05 (s, 2H), 7.45 (m, 4H), 7.65 (d, 2H), 8.00 (br, 3H, NH3+), 8.18 (d, 2H). 8.35 (br, 3H, NH3+), 9.14 (br, 1H, NH) LCMS: 595.49 [M+H] HPLC: Purity 94.09% at 8.68 minutes.

[0217] [Example 20] Preparation of (1S)-2-carboxy-1-[(2-{[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methoxy}-2-oxoethyl)carbamoyl]ethane-1-aminium chloride (compound 22) [ka]

[0218] (Step 1) Preparation of 2-{[(tert-butoxy)carbonyl]amino}benzyl acetate (compound Q2) To a stirred solution of 2-{[(tert-butoxy)carbonyl]amino}acetic acid (compound E1; 2.0 g, 11.42 mmol) in DCM (40 mL, 20 V), DMAP (130 mg, 1.14 mmol) was added at 0-5°C, followed by EDC.HCl (2.65 g, 17.12 mmol), and the resulting mixture was stirred for 5 minutes. Phenylmethanol (Q1; 1.35 g, 12.55 mmol) was added to the above reaction mixture at 0-5°C, and the mixture was stirred at ambient temperature (25-30°C) for 16 hours.

[0219] The reaction product was diluted with dichloromethane (200 mL), washed with water (2 × 100 mL), and concentrated under reduced pressure. The residue obtained by evaporating the solvent was purified by silica gel flash column chromatography to obtain compound Q2 as a white solid (2 g, yield 66%).

[0220] (Step 2) Preparation of 2-benzyl aminoacetate (compound Q3) To a stirred solution of benzyl 2-{[(tert-butoxy)carbonyl]amino}acetate (compound Q2; 2g, 7.54 mmol) in dichloromethane (30 mL, 15V), trifluoroacetic acid (4 mL, 2V) was added at 0-5°C. The reaction mixture was stirred at ambient temperature (25-30°C) for 3 hours. The reaction mixture was concentrated to remove volatiles, and the resulting residue was powdered with diethyl ether to obtain compound Q3 as a white solid (1.8 g, yield 85%).

[0221] (Step 3) Preparation of (3S)-3-{[2-(benzyloxy)-2-oxoethyl]carbamoyl}-3-{[(tert-butoxy)carbonyl]amino}propanoate tert-butyl (compound Q5) To a stirred solution of (2S)-4-(tert-butoxy)-2-{[(tert-butoxy)carbonyl]amino}-4-oxobutanoic acid (compound Q4; 4.37 g, 15.1 mmol) in DCM (20 mL, 20 V), DMAP (140 mg, 1.21 mmol) was added at 0-5°C, followed by EDC.HCl (3.23 g, 16.95 mmol), and the mixture was stirred for 5 minutes. 2-benzyl aminoacetate (compound Q3; 1.0 g, 6.05 mmol) and DIPEA (2.96 mL, 18.16 mmol) were added to the reaction mixture at 0-5°C, and the reaction mixture was stirred at ambient temperature (25-30°C) for 16 hours. The reaction product was diluted with dichloromethane (400 mL), washed with water (2 × 200 mL), and concentrated under reduced pressure. The residue obtained by evaporating the solvent was purified by silica gel flash column chromatography to obtain compound Q5 as a pale yellow wax (1 g, yield 64%).

[0222] (Step 4) Preparation of 2-[(2S)-4-(tert-butoxy)-2-{[(tert-butoxy)carbonyl]amino}-4-oxobutanamide]acetic acid (compound Q6) To a solution of tert-butyl (3S)-3-{[2-(benzyloxy)-2-oxoethyl]carbamoyl}-3-{[(tert-butoxy)carbonyl]amino}propanoate (compound Q5; 1.0 g, 2.29 mmol) in ethyl acetate (30 mL, 30 V), Pd / C (0.1 g, 10% w / w) was added, and the resulting mass was hydrogenated in a parr apparatus (5 kg H2 pressure) for 3 hours. The reaction product was filtered through a celite pad and concentrated under reduced pressure. The resulting residue was powdered with diethyl ether to obtain compound Q6 as a white solid (0.67 g, yield 84%).

[0223] (Step 5) Preparation of (3S)-3-{[(tert-butoxy)carbonyl]amino}-3-[(2-{[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methoxy}-2-oxoethyl)carbamoyl]propanoate tert-butyl (compound Q7) To a solution of 2-[(2S)-4-(tert-butoxy)-2-{[(tert-butoxy)carbonyl]amino}-4-oxobutanamide]acetic acid (compound Q6; 0.64 g, 1.87 mmol) in THF (7 mL, 20 V), DMAP (0.052 g, 0.42 mmol) and then DCC (0.43 g, 2.13 mmol) were added at 0-5°C and the mixture was stirred for 20 minutes. Compound 2 (0.35 g, 0.85 mmol) was added to the reaction mixture at 0-5°C and the reaction mixture was stirred at ambient temperature (25-30°C) for 16 hours. The reaction mixture was filtered, the filtrate was concentrated, and the filtrate was purified by reverse-phase column chromatography to obtain compound Q7 as a white solid (0.5 g, yield 79%).

[0224] (Step 6) Preparation of (1S)-2-carboxy-1-[(2-{[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methoxy}-2-oxoethyl)carbamoyl]ethane-1-aminium trifluoroacetate (compound Q8) To a stirred solution of (3S)-3-{[(tert-butoxy)carbonyl]amino}-3-[(2-{[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methoxy}-2-oxoethyl)carbamoyl] tert-butyl propanoate (compound Q7; 0.5 g, 0.67 mmol) in dichloromethane (7.5 mL), trifluoroacetic acid (2 mL, 4.0 vol) was added at 0-5°C, and the mixture was stirred at ambient temperature (25-30°C) for 4 hours. The reaction product was concentrated, volatiles were removed, and the resulting residue was powdered with diethyl ether to obtain compound Q8 as a white solid (0.25 g, yield 53%).

[0225] (Step 7) Preparation of (1S)-2-carboxy-1-[(2-{[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methoxy}-2-oxoethyl)carbamoyl]ethane-1-aminium chloride (compound 22) To a stirred solution of (1S)-2-carboxy-1-[(2-{[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methoxy}-2-oxoethyl)carbamoyl]ethane-1-aminium trifluoroacetate (compound Q8; 0.25 g, 0.35 mmol) in THF (1.25 mL, 5.0 vol), HCl in diethyl ether (5.0 mL, 20 vol) was added at 0-5°C, and the reaction mixture was stirred at the same temperature for 20 minutes. The reaction product was concentrated to remove volatiles, and the resulting residue was powdered with diethyl ether to obtain compound 22 as a white solid (200 mg, yield 90%). 1H NMR (400 MHz, DMSO) δ (ppm): 2.85 (m, 2H), 4.07 (m, 3H), 4.83 (s, 2h), 6.04 (s, 2H), 7.44 (m, 4H), 7.65 (d, 2H), 7.63 (d, 2H), 8.14 (d, 2H), 8.25 (br, 3H, NH3+), 9.01 (s, 1H), 12-14 (br, 1H, -COOH) LCMS: 582.3 M+H] HPLC: Purity 95.26% at 11.43 minutes.

[0226] [Example 21] Preparation of Azanium ({[(4S)-4-Azanium-4-[(2-{[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methoxy}-2-oxoethyl)carbamoyl]butyl]amino})methylidene]azanium trichloride (Compound 23) [ka]

[0227] (Step 1) Preparation of (2S)-2-{[(tert-butoxy)carbonyl]amino}-5-{[(1E)-{[(tert-butoxy)carbonyl]amino}({[(tert-butoxy)carbonyl]imino})methyl]amino}pentanoic acid (compound R2) To a solution of (2S)-2-amino-5-carbamimidoamidopentanoic acid (compound R1; 8.8 g, 50.51 mmol) in t-butanol (158 mL, 18V) and water (158 mL, 18V), NaOH (7.07 g, 176.80 mmol) and Boc2O (46.42 mL, 202.06 mmol) were added at 0°C. The reaction mixture was stirred at ambient temperature (25-30°C) for 48 hours. Volatile substances were evaporated from the reaction product, and the pH was carefully acidified to 3-4 with saturated citric acid solution. The product was extracted with ethyl acetate, the extract was dried over Na2SO4, filtered, evaporated under vacuum, and purified by reverse-phase column chromatography to obtain compound R2 as a white solid (3.7 g, yield 15%).

[0228] (Step 2) Preparation of 2-[(2S)-2-{[(tert-butoxy)carbonyl]amino}-5-{[(1E)-{[(tert-butoxy)carbonyl]amino}({[(tert-butoxy)carbonyl]imino})methyl]amino}pentanamide]benzyl acetate (compound R3) To a stirred solution of (2S)-2-{[(tert-butoxy)carbonyl]amino}-5-{[(1E)-{[(tert-butoxy)carbonyl]amino}({[(tert-butoxy)carbonyl]imino})methyl]amino}pentanoic acid (compound R2; 1.33 g, 2.15 mmol) in THF (8 mL, 20 V), DMAP (0.052 g, 0.43 mmol) was added at 0-5°C, followed by EDC.HCl (0.54 g, 2.86 mmol), and the mixture was stirred for 20 minutes. To the above reaction mixture, benzyl aminoacetate (compound Q3; 0.4 g, 1.43 mmol) in TEA (0.6 mL, 4.30 mmol) was added at 0-5°C, and the reaction mixture was stirred at ambient temperature (25-30°C) for 16 hours. The reaction product was diluted with water (100 mL), extracted with dichloromethane (200 mL), dried, and evaporated under vacuum. The residue was purified by silica gel column chromatography to obtain compound R3 as a white solid (0.6 g, yield 67%).

[0229] (Step 3) Preparation of 2-[(2S)-2-{[(tert-butoxy)carbonyl]amino}-5-{[(1E)-{[(tert-butoxy)carbonyl]amino}({[(tert-butoxy)carbonyl]imino})methyl]amino}pentanamide]acetic acid (compound R4) To a solution of 2-[(2S)-2-{[(tert-butoxy)carbonyl]amino}-5-{[(1E)-{[(tert-butoxy)carbonyl]amino}({[(tert-butoxy)carbonyl]imino})methyl]aminopentanamide]benzyl acetate (compound R3; 0.6 g, 0.96 mmol) in ethyl acetate (6 mL, 10 V), Pd / C (0.06 g, 10% w / w) was added, and the mixture was hydrogenated in a parr apparatus (5 kg H2 pressure) for 3 hours. The reaction product was filtered through a celite bed, and the filtrate was concentrated under reduced pressure. The residue was washed with ether to obtain compound R4 as an off-white solid (0.43 g, yield 84%).

[0230] (Step 4) Preparation of [5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methyl 2-[(2S)-2-{[(tert-butoxy)carbonyl]amino}-5-{[(1E)-{[(tert-butoxy)carbonyl]amino}({[(tert-butoxy)carbonyl]imino})methyl]amino}pentanamide]acetate (compound R5) To a stirred solution of 2-[(2S)-2-{[(tert-butoxy)carbonyl]amino}-5-{[(1E)-{[(tert-butoxy)carbonyl]amino}({[(tert-butoxy)carbonyl]imino})methyl]aminopentanamide]acetic acid (compound R4; 0.41 g, 0.77 mmol) in THF (3.2 mL, 20 V), DMAP (0.014 g, 0.11 mmol) and then DCC (0.18 g, 0.89 mmol) were added at 0-5°C and the mixture was stirred for 20 minutes. Compound 2 (0.16 g, 0.38 mmol) was added to the above reaction mixture at 0-5°C and the mixture was further stirred at ambient temperature (25-30°C) for 16 hours. The reaction mixture was filtered, the filtrate was concentrated, and the compound R5 was purified by reverse-phase column chromatography to obtain compound R5 as a white solid (0.25 g, yield 69%).

[0231] (Step 5) Preparation of [Azaniumyl ({[(4S)-4-Azaniumyl-4-[(2-{[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methoxy}-2-oxoethyl)carbamoyl]butyl]amino})methylidene]azanium trichloride (compound 23) To a stirred solution of [5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methyl 2-[(2S)-2-{[(tert-butoxy)carbonyl]amino}-5-{[(1E)-{[(tert-butoxy)carbonyl]amino}({[(tert-butoxy)carbonyl]imino})methyl]aminopentanamide]acetate (compound R5; 0.2 g, 0.21 mmol) in THF (1.0 mL, 5.0 vol), HCl in diethyl ether (4.0 mL, 20 vol) was added at 0-5°C, and the reaction mixture was stirred at the same temperature for 4 hours. The reaction product was concentrated to remove volatiles, and the resulting residue was powdered with diethyl ether to obtain compound 23 as a white solid (90 mg, yield 60%). 1H NMR (400 MHz, DMSO) δ (ppm):1.45 (br, 3H), 1.78 (br, 4H), 3.14 (m, 2H), 3.88 (m, 1H), 4.05 (m, 2H), 4.84 (s, 2H), 6.04 (s, 2H), 7.45 (q, 5H), 7.65 (dd, 2H), 7.83 (t, 1H), 8.18 (d, 2H), 8.35 (br, 4H), 9.20 (t, 1H) LCMS: 623.3 [M+H] HPLC: Purity 92.07% at 8.52 minutes.

[0232] [Example 22] Stability of the compound A 5 μM solution of compound 1 and compound 4 (i.e., citrate of compound 3) in HCl buffer (pH: 1.20) was prepared using 5 mM DMSO stock solution. The solution was incubated at 37°C for 120 minutes while shaking at 400 rpm using a thermomixer.

[0233] At intervals of 0, 15, 30, 60, and 120 minutes, aliquots of a sufficient amount of compound solution were taken, diluted, and analyzed to measure the stability of compounds 1 and 3 in acidic solutions.

[0234] (result) The results of the stability experiment are shown in Table 3 below.

[0235] As a result, it was found that compound 3 has higher stability than compound 1 in an acidic solution (i.e., pH = 1.2).

[0236] [Table 2]

[0237] [Example 23] Activation of AMPK (Culture and compound treatment of INS-1E insulinoma cells) INS-1E cells were cultured as described in Steneberg et al., JCI Insight. 2018;3(12):e99114. https: / / doi.org / 10.1172 / jci.insight.99114, and 5% fetal bovine serum was used instead of 10% fetal bovine serum when plating for treatment.

[0238] Compound 3 was dissolved in DMSO at a concentration of 10 mM and frozen at -20°C.

[0239] INS-1E cells were treated in serum-free medium for 4 hours with increasing doses of compound 3, following the method described in Steneberg et al., JCI Insight. 2018;3(12):e99114. https: / / doi.org / 10.1172 / jci.insight.99114.

[0240] [Western blot analysis] Western blot analysis of INS-1E cells was performed as described in Steneberg et al., JCI Insight. 2018;3(12):e99114. https: / / doi.org / 10.1172 / jci.insight.99114. Cell lysates were passed through a 30-gauge needle and centrifuged approximately eight times at +4°C for 10 minutes at 14000 rpm. Quantitative values ​​of AMPKα and phosphorylated-T172 AMPKα were normalized to β-actin quantification values.

[0241] (result) The results of the Western blot analysis are shown in Table 4 below, and are illustrated in Figure 1.

[0242] The results showed that compound 3 dose-dependently increased phosphorylated-T172 AMPK in cultured INS-1E cells. Therefore, compound 3 is an AMPK agonist.

[0243] [Table 3]

[0244] [Example 24] Pharmacokinetic study of compounds 1 and 3 administered orally in Sprague Dawley rats (Formulation of compound 1) 10.0 mL of a 2% w / v methylcellulose solution in phosphate buffer (pH 7.5) was added to a 100 mL conical flask along with 1 g of 2 mm glass beads. The solution was vigorously stirred with a magnetic stirrer. 100 mg of compound 1 was slowly added to the solution, and the solution was vigorously stirred for approximately 1 hour. The pH of the preparation was measured to be 7.49. Each preparation was freshly prepared before administration to the animals. The final concentration of compound 1 in the preparation was 10 mg / mL. The preparation was administered at a dose of 5 mL / kg body weight.

[0245] (Formulation of compound 3) Six mL of a 2% w / v methylcellulose solution in phosphate buffer (pH 7.5) was added to a 25 mL conical flask along with 1 g of 2 mm glass beads. The solution was vigorously stirred with a magnetic stirrer. 78 mg of compound 3 was slowly added to the solution, and the solution was vigorously stirred for approximately 1 hour. The pH of the preparation was measured to be 7.41. Each preparation was freshly prepared before administration to the animals. The final concentration of compound 3 in the preparation was 13 mg / mL, which corresponds to 10 mg / mL of compound 1. The preparation was administered at a dose of 5 mL / kg body weight.

[0246] (Selection of dosage and justification for selection) For equimolar dose comparison, doses of 50 mg / kg and 65 mg / kg b.w. were selected for compound 1 and compound 3, respectively.

[0247] (Administration) Healthy adult male Sprague Dawley rats aged 8-10 weeks were used in the experiment after a minimum of 3 days of acclimatization. The fed animals were orally administered either compound 1 or compound 3 at doses of 50 mg / kg or 65 mg / kg body weight, respectively, via gastric tube feeding.

[0248] (Blood sampling) Under mild isoflurane anesthesia, blood samples were collected within 72 hours post-administration using a retroorbital puncture method with a capillary tube, in pre-labeled tubes containing an anticoagulant (K2EDTA: 2 mg / mL blood). Details are shown in Table 6. When blood was collected at multiple time points, the right and left eyes were used alternately. The collected blood samples were centrifuged at 6000 rpm at 4°C for 10 minutes to separate the plasma samples, which were then stored at -80°C until analysis.

[0249] (result) The results of the single-dose oral pharmacokinetic study in rats are shown in Tables 5 and 6 below, and are illustrated in Figure 2. In Table 5, C max , T max AUC last AUC inf、 AUC extrap , T 1 / 2 and MRT last The value is for 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide (compound 1) detected after administration of both compound 1 and compound 3.

[0250] As a result, when compound 3 was administered, the systemic exposure to 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide increased tenfold, surprisingly, compared to compound 1 (the Cmax after administration of compound 1 was 19.60 μg per mL for compound 1, compared to 207.00 μg per mL for compound 1 when compound 3 was administered). Therefore, systemic exposure to 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide is increased by administering the compound in the form of the compound of the present invention.

[0251] [Table 4]

[0252] [Table 5]

[0253] [Example 25] Single oral pharmacokinetic study of compounds 5, 6, 7, 9, 10, 13, 14, and 16 in Sprague Dawley rats (Compound formulation) The formulation was prepared according to the method described in Example 19.

[0254] (Selection of dosage and justification for selection) For equimolar dose comparison, a dose equivalent to 50 mg / kg of compound 1 was selected.

[0255] (Administration and blood sampling) Administration and blood collection were carried out as described in Example 19.

[0256] (result) The results of the single-dose oral pharmacokinetic study in rats are shown in Table 7 and below, and are illustrated in Figure 3. In Table 7, C max , T max AUC last AUC inf、 AUC extrap and T 1 / 2 The value shown is that of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide (compound 1) detected after compound administration.

[0257] The results showed that administration of compounds 5, 6, 7, 9, 10, 13, 14, 16, and 17 resulted in mean maximum plasma concentrations (Cmax) of 4-chloro-N-[2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazole-5-yl]benzamide ranging from 71 to 137 μg / mL.

[0258] [Table 6]

[0259] [Table 7]

[0260] [Example 26] Stability of Compound 17 The stability of compound 17 was evaluated in buffers at pH 1.20, 7.40, and 9.20. The pH 1.20 test was performed using HCl buffer. Phosphate buffer was used for the pH 7.40 medium, and alkaline borate buffer was used for the pH 9.20 medium.

[0261] (HCl buffer) Potassium chloride (1.491 g) was dissolved in 100 mL of ultrapure water type-1 to obtain a 0.2 M KCl solution. Approximately 50 mL of the 0.2 M KCl was placed in a beaker, and the pH was adjusted to 1.20 using a 0.2 M HCl solution. The final volume was increased to 200 mL using ultrapure water type-1, and the solution was stored at room temperature (22 ± 3 °C).

[0262] (Phosphate buffer) Monobasic potassium phosphate (2.722 g) was dissolved in 100 mL of ultrapure water type-1 to obtain a 0.2 M solution. Approximately 50 mL of the 0.2 M solution was placed in a beaker, and the pH was adjusted to 7.40 using 0.2 M NaOH solution. The final volume was increased to 200 mL using ultrapure water type-1, and the solution was stored at room temperature (22 ± 3 °C).

[0263] (Alkaline borate buffer) Boric acid (1.237 g) and potassium chloride (1.491 g) were dissolved in 100 mL of ultrapure water type-1 to obtain a 0.2 M solution. Approximately 50 mL of the 0.2 M solution was placed in a beaker, and the pH was adjusted to 9.20 using 0.2 M NaOH solution. The final volume was increased to 200 mL using ultrapure water type-1, and the solution was stored at room temperature (22 ± 3 °C).

[0264] A 5 μM solution of compound 17 in each buffer was prepared using 5 mM DMSO stock solution. These solutions were incubated at 22 ± 3 °C for 120 minutes while being shaken at 400 rpm using a thermomixer.

[0265] At 0, 15, 30, 60, and 120-minute intervals, aliquots of a sufficient amount of the compound solution were taken, diluted, and analyzed to measure the stability of compound 17 and the abundance of compound 1 in each solution. The amount of compound 1 present was compared to the amount expected assuming complete conversion from compound 17 to compound 1.

[0266] (result) The results of the stability experiment are shown in Table 9 below.

[0267] [Table 8]

[0268] As a result, it was found that compound 17 exhibits maximum stability in a low pH environment (pH=1.2), but decomposes rapidly at pH 7.40 or higher.

[0269] [Example 27] Additional stability results The stability of compounds 3, 6, 9, 20, and 23 was tested using the same method as described above for compound 17. The amounts of each compound remaining at 15 and 30 minutes are shown in Table 10.

[0270] [Table 9]

[0271] Compounds 17 and 23 were found to degrade rapidly at pH 7.40 and be relatively stable at pH 1.2. These data suggest that stability at pH 7.4 is actually higher when the amino acid side chain is lipophilic (e.g., alkyl, see compound 20). Stability was lower when the side chain contained H or -NH2 (glycine or lysine-based). When aspartic acid was used (compound 17), stability became very low at higher pH levels. While this compound may remain intact in the stomach, it may be hydrolyzed in the intestines. [Brief explanation of the drawing]

[0272] [Figure 1] This is a Western blot image showing that compound 3 increases AMPK phosphorylation in a dose-dependent manner. [Figure 2] This is a comparative result of oral pharmacokinetic studies using compound 1 and compound 3. [Figure 3] This is a comparison of oral pharmacokinetic studies using compounds 5, 6, 7, 9, 10, 13, 14, 16, and 17.

Claims

1. Compounds of formula I or their pharmaceutically acceptable salts or solvates: 【Chemistry 1】 R １ and R ２ These are independently hydrogen, the side chains of protein constituent amino acids, and -OH, -NH ２ and -C(O)NH ２ C, which is optionally substituted with one or more substituents selected from the group consisting of １－６ Selected from the group consisting of alkyl groups; R ３ and R ４ are independently selected from hydrogen, C optionally substituted by a phenyl group １－４ alkyl, and A １ ; or are selected from the group consisting of R １ (or R ２ ) and R ３ (or R ４ ) together with the carbon and nitrogen atoms to which they are bonded, form a 4- to 6-membered heterocycloalkyl group; A １ teeth, 【Chemistry 2】 or 【Transformation 3】 This represents, A ２ C is optionally substituted with hydrogen or a phenyl group. １－４ Alkyl, 【Chemistry 4】 or 【Transformation 5】 This represents, Each A ３ This is a C atom that is independently substituted with hydrogen or a phenyl group. １－４ Represents alkyl; and R ５ａ and R ５ｂ These are independently hydrogen, the side chains of protein constituent amino acids, and -OH, -NH ２ and -C(O)NH ２ C, which is optionally substituted with one or more substituents selected from the group consisting of １－６ Selected from the group consisting of alkyl groups.

2. R ２ It is hydrogen, and furthermore, R １ is hydrogen, or a side chain of a protein-constituting amino acid selected from the group consisting of Glu, Arg, His, Lys, Ser, Thr, Asn, Gln, Cys, Sec, Ala, Ile, Leu, Met, Phe, Trp, Tyr, Asp, and Val; or, R ２ It is hydrogen, and furthermore, R １ and R ３ The compound according to claim 1, wherein they form a pyrrolidine ring together with the carbon and nitrogen atoms to which they are bonded.

3. R ２ It is hydrogen, and furthermore, R １ is either hydrogen, or a side chain of a protein constituent amino acid selected from the group consisting of Arg, His, Lys, and Trp, or R ２ It is hydrogen, and furthermore, R １ and R ３ The compound according to claim 1, wherein they form a pyrrolidine ring together with the carbon and nitrogen atoms to which they are bonded.

4. R ２ It is hydrogen, and furthermore, R １ The compound according to claim 1, wherein is the side chain of aspartic acid.

5. R ３ and R ４ However, independently, C is optionally substituted with hydrogen and one or more phenyl groups. １－３ Alkyl, 【Transformation 6】 and 【Transformation 7】 A group consisting of the following is selected, and further, R ５ａ The compound according to claim 1, wherein is hydrogen or a side chain of a protein constituent amino acid.

6. R ３ and R ４ However, independently, C is optionally substituted with hydrogen and one or more phenyl groups. １－３ Alkyl, 【Transformation 8】 and 【Chemistry 9】 Selected from the group consisting of, R １ and R ３ However, together with the carbon and nitrogen atoms to which they are bonded, they form a pyrrolidine ring, and furthermore, R ４ However, C is optionally substituted with hydrogen and one or more phenyl groups. １－３ Alkyl, 【Chemistry 10】 and 【Chemistry 11】 Selected from the group consisting of, R ５ａ The compound according to claim 1, wherein the side chain is a protein constituent amino acid selected from the group consisting of Phe, Val, Lys, Asp, and Arg.

7. R ３ and R ４ However, independently, C is optionally substituted with hydrogen and one or more phenyl groups. １－３ A compound according to claim 1, selected from the group consisting of alkyl groups.

8. R ３ and R ４ The compound according to claim 1, wherein the compound is independently selected from the group consisting of hydrogen and methyl.

9. R ２ is hydrogen, R １ R is a side chain of a protein constituent amino acid selected from the group consisting of hydrogen, or Glu, Arg, His, Lys, Ser, Thr, Asn, Gln, Cys, Sec, Ala, Ile, Leu, Met, Phe, Trp, Tyr, Asp, and Val. ３ and R ４ However, independently, C is optionally substituted with hydrogen and one or more phenyl groups. １－３ Alkyl, 【Chemistry 12】 and 【Chemistry 13】 Selected from the group consisting of, R ２ is hydrogen, R １ and R ３ However, together with the carbon and nitrogen atoms to which they are bonded, they form a pyrrolidine ring, and furthermore, R ４ However, C is optionally substituted with hydrogen and one or more phenyl groups. １－３ Alkyl, 【Chemistry 14】 and 【Chemistry 15】 Selected from the group consisting of, R ５ａ The compound according to claim 1, wherein the side chain is hydrogen or a protein constituent amino acid selected from the group consisting of Glu, Arg, His, Lys, Ser, Thr, Asn, Gln, Cys, Sec, Ala, Ile, Leu, Met, Phe, Trp, Tyr, Asp, and Val.

10. The above compound, Dimethylaminoacetic acid 5-[(Z)-4-chloro-benzoylimino]-2-(4-chloro-benzyl)-3-oxo-[1,2,4]thiadiazolidine-4-ylmethyl ester; Citrate of dimethylaminoacetic acid 5-[(Z)-4-chlorobenzoylimino]-2-(4-chlorobenzyl)-3-oxo-[1,2,4]thiadiazolidine-4-ylmethyl ester; 2-({[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methoxy}carbonyl)pyrrolidine-1-ium trifluoroacetate; 6-{[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methoxy}-6-oxohexane-1,5-bis(aminium)di-trifluoroacetate; (6-Azaniumyl-1-{[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methoxy}-1-oxohexane-2-yl)dimethylazanium di-trifluoroacetate; (2-{[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methoxy}-2-oxoethyl)(methyl)azanium trifluoroacetate; 2-{[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methoxy}-2-oxoethane-1-aminium trifluoroacetate; 2-({[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methoxy}carbonyl)-1-methylpyrrolidine-1-ium trifluoroacetate; 1-{[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methoxy}-1-oxopropane-2-aminium trifluoroacetate; 1-{[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methoxy}-3-methyl-1-oxopentane-2-aminium trifluoroacetate; [5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methyl 2-(dimethylamino)-3-phenylpropanoate; 1-{[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methoxy}-1-oxo-3-phenylpropane-2-aminium trifluoroacetate; 1-{[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methoxy}-4-methyl-1-oxopentane-2-aminium trifluoroacetate; 1-{[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methoxy}-3-hydroxy-1-oxopropane-2-aminium trifluoroacetate; 3-Carboxy-1-{[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methoxy}-1-oxopropane-2-aminium trifluoroacetate; [5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methyl 2-[(2S)-2-azaniumyl-3-phenylpropanamide]acetate chloride; [5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methyl (2S)-2-[(2S)-2-azaniumyl-3-methylbutanamide]-3-methylbutanoate chloride; [5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methyl (2R)-2-[(2R)-2-azaniumyl-3-methylbutanamide]-3-methylbutanoate chloride; [5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methyl 2-[(2S)-2,6-diazaniumylhexanamide]acetate dichloride; (1S)-2-carboxy-1-[(2-{[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methoxy}-2-oxoethyl)carbamoyl]ethane-1-aminium chloride; and [Azanium ({[(4S)-4-Azanium-4-[(2-{[5-(4-chlorobenzamide)-2-[(4-chlorophenyl)methyl]-3-oxo-1,2,4-thiadiazolidine-4-yl]methoxy}-2-oxoethyl)carbamoyl]butyl]amino})methylidene]azanium trichloride A compound according to claim 1, selected from the group consisting of the following.

11. A pharmaceutical formulation comprising a compound defined in any one of claims 1 to 10 or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable excipient.

12. A pharmaceutical preparation as defined in claim 11, for use in pharmaceuticals.

13. A pharmaceutical formulation as defined in claim 11, for use in the treatment of a disorder or health condition that is improved by activation of AMPK.

14. Use of a compound of formula I as defined in any one of claims 1 to 10 or a pharmaceutically acceptable salt or solvate thereof in the manufacture of a pharmaceutical for the treatment of a disorder or health condition that is improved by activation of AMPK.

15. A pharmaceutical formulation comprising a compound defined in any one of claims 1 to 10 or a pharmaceutically acceptable salt or solvate thereof, for use in the treatment of a disorder or health condition that is improved by activation of AMPK, administered to a subject in need.

16. The pharmaceutical formulation according to claim 13, wherein the disorder or health condition improved by activation of AMPK is selected from the group consisting of cardiovascular disease (such as heart failure), diabetic nephropathy, type 2 diabetes, insulin resistance, non-alcoholic fatty liver disease, non-alcoholic steatohepatitis, pain, opioid addiction, obesity, cancer, inflammation (including chronic inflammatory diseases), autoimmune diseases, osteoporosis, intestinal diseases, and hyperinsulinemia associated with obesity or cardiovascular disease.

17. (i) Compound of formula III and 【Chemistry 16】 Reaction with the compound of formula IV, 【Chemistry 17】 or (ii) Compound of formula V and [Chemistry 18] This includes reactions with compounds of formula IV, 【Chemistry 19】 R １ and R ２ However, as defined in any one of claims 1 to 10; and, R ３ and R ４ A method for preparing a compound as defined in any one of claims 1 to 10, which is either as defined in any one of claims 1 to 10, or independently represents a protecting group.

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