(2S)-N-[(1S)-1-cyano-2-phenylethyl]-1,4-oxazepan-2-carboxamide as a dipeptidyl peptidase 1 inhibitor

Abstract

We provide certain (2S)-N-[(1S)-1-cyano-2-phenylethyl]-1,4-oxazepan-2-carboxamide compounds that inhibit dipeptidyl peptidase 1 (DPP1; EC3.4.14.1) activity. [Solution] A compound represented by formula (I) or a pharmaceutically acceptable salt thereof is provided. TIFF2026065113000111.tif4164 The usefulness of the compounds in the treatment and / or prevention of clinical conditions, including respiratory diseases such as asthma and chronic obstructive pulmonary disease (COPD), their use in treatment, pharmaceutical compositions containing them, and methods for producing the compounds are also provided.

Description

[Technical Field]

[0001] The present technical field relates to certain (2S)-N-[(1S)-1-cyano-2-phenylethyl]-1,4-oxazepan-2-carboxamide compounds (including pharmaceutically acceptable salts thereof) that inhibit dipeptidyl peptidase 1 (DPP1; EC3.4.14.1) activity, their usefulness in the treatment and / or prevention of clinical conditions including respiratory diseases such as asthma and chronic obstructive pulmonary disease (COPD), their use in treatment, pharmaceutical compositions containing them, and methods for producing said compounds. [Background technology]

[0002] Dipeptidyl peptidase 1 (DPP1; EC3.4.14.1), also known as cathepsin C, is a lysosomal cysteine ​​protease belonging to the papain family with a molecular weight of 200 kDa. DPP1 was first discovered in 1948 by Gutman and Fruton (Non-Patent Literature 1); however, the cDNA of the human enzyme was first described in 1995 (Non-Patent Literature 2). DPP1 is the only member of the papain family that functions as a tetramer consisting of four identical subunits. Each subunit consists of an N-terminal fragment, a heavy chain, and a light chain (Non-Patent Literature 3).

[0003] DPP1 is constitutively expressed in many tissues, with the highest levels in the lungs, kidneys, liver, and spleen. DPP1 catalyzes the removal of dipeptides from the N-terminus of polypeptide substrates with broad specificity. Recent data suggest that, in addition to being an important enzyme in lysosomal proteolysis, DPP1 also functions as a key enzyme in the activation of granular serine proteases in cytotoxic T lymphocytes and natural killer cells (granzymes A and B), mast cells (chymase and tryptase), and neutrophils (cathepsin G, neutrophil elastase, and proteinase 3).

[0004] Mast cells are found in many tissues, but the majority reside along the endothelial layer of the body, such as in the skin, respiratory tract, and gastrointestinal tract. In humans, two types of mast cells have been identified: the T-type, which expresses only tryptase, and the MC-type, which expresses both tryptase and chymase. In humans, T-type mast cells are mainly found in alveolar tissue and intestinal mucosa, while TC-type cells are mainly found in the skin and conjunctiva. Tryptase and chymase are thought to be important mediators in allergic diseases, involved in the processes of inflammation, bronchoconstriction, and mucus secretion.

[0005] Neutrophils play a crucial role in the host's defense against pathogen invasion. Produced in the bone marrow, neutrophils are fully mature when released into the circulatory system to serve as the first line of cellular defense. Pro-inflammatory mediators and chemotactic attractants activate neutrophils, drawing them to the site of infection, where they phagocytose bacteria and attack them with a arsenal of antibacterial compounds employing both oxidative and non-oxidative attack mechanisms. Neutrophil elastase, a potent serine protease, is clearly one of these antibacterial compounds involved in the destruction of bacteria. Neutrophil elastase is released into phagolysosomes surrounding microorganisms and begins to destroy them. Neutrophil elastase can attack the outer membrane protein OmpA of Gram-negative bacteria, helping to directly kill the pathogen by disrupting its membrane, while simultaneously allowing other antibacterial compounds to enter the pathogen. In addition, neutrophil elastase can assist in the processing of other antibacterial compounds, and, as in the case of cathelicidin, can convert these antibacterial compounds from an inactive propeptide to an active state.

[0006] Nevertheless, neutrophil elastase can also cause problems for its host. It is one of the most destructive enzymes in the body, capable of degrading extracellular matrix proteins (including collagen, proteoglycans, fibronectin, platelet receptors, complement receptors, thrombomodulin, pulmonary surfactant, and cadherins) and important plasma proteins (including coagulation and complement factors, immunoglobulins, several proteases, and protease inhibitors). Endogenous protease inhibitors such as α1-antitrypsin tightly control neutrophil elastase activity under physiological conditions. However, neutrophil elastase can evade control at sites of inflammation, and once uncontrolled, it can trigger the release of pro-inflammatory cytokines such as interleukin-6 and interleukin-8, leading to acute lung injury. It can even impair host defense against infection by degrading phagocytic cell surface receptors and opsonins. Its negative role is explained by its involvement in tissue destruction and inflammation that characterize many diseases, including hereditary emphysema, chronic obstructive pulmonary disease, cystic fibrosis, adult respiratory distress syndrome, ischemic-reperfusion injury, and rheumatoid arthritis.

[0007] There is strong evidence linking tryptases and chymases to many mast cell-mediated allergic, immunological, and inflammatory diseases. The fact that neutrophil elastase, cathepsin G, and proteinase 3 also appear to play important roles in these types of diseases suggests that DPPIs are a reasonable therapeutic target due to their central role in the activation of these proteases (Non-Patent Literature 4; Non-Patent Literature 5).

[0008] Patent Document 1 relates to certain nitrile derivatives and their use as DPP1 inhibitors.

[0009] Patent document 2 relates to peptidylnitrile and their use as DPP1 inhibitors.

[0010] Patent document 3 relates to α-aminoamidonitriles and their use as DPP1 inhibitors.

[0011] Patent document 4 relates to peptidylnitrile compounds and their use as DPP1 inhibitors.

[0012] Patent document 5 relates to N-[1-cyano-2-(phenyl)ethyl]-2-azabicyclo[2.2.1]heptan-3-carboxamide and their use as DPP1 inhibitors.

[0013] Patent documents 6 and 7 relate to β-aminoamidonitriles having inhibitory activity against cysteine ​​proteases. [Prior art documents] [Patent Documents]

[0014] [Patent Document 1] International Publication No. 2004 / 110988 [Patent Document 2] International Publication No. 2009 / 074829 [Patent Document 3] International Publication No. 2010 / 128324 [Patent Document 4] International Publication No. 2012 / 119941 [Patent Document 5] International Publication No. 2013 / 041497 [Patent Document 6] International Publication No. 2001 / 096285 [Patent Document 7] International Publication No. 2003 / 048123 [Non-patent literature]

[0015] [Non-Patent Document 1] J Biol Chem, 174, 851-858 [Non-Patent Document 2] Paris et al. 1995, FEBS Lett, 369, 326-330 [Non-Patent Document 3] Dolenc et al. 1995, J Biol Chem, 270, 21626-21631 [Non-Patent Document 4] Adkison et al. 2002, J Clin Invest, 109, 363-271 [Non-Patent Document 5] Pham et al. 2004, J Immunol, 173, 7277-7281 [Overview of the Initiative] [Problems that the invention aims to solve]

[0016] No amide nitrile compounds having a β-amino acid in the form of the described (2S)-N-[(1S)-1-cyano-2-phenylethyl]-1,4-oxazepane-2-carboxamide compound have been disclosed. We have now found that such compounds possess potent DPP1 activity and / or a desirable pharmacological activity profile (e.g., reduced risk of binding to elastin-rich tissues such as the aorta).

[0017] overview A compound that is an inhibitor of dipeptidyl peptidase 1 (DPP1), the use of the compound as a pharmaceutical, a pharmaceutical composition containing the compound, and a synthetic route for producing the compound are provided. [Means for solving the problem]

[0018] According to the first aspect, equation (I): [ka] [In the formula, R 1 teeth, [ka] and; R 2 is selected from hydrogen, F, Cl, Br, OSO2C 1-3 alkyl or C 1-3 alkyl; R 3 is selected from hydrogen, F, Cl, Br, CN, CF3, SO2C 1-3 alkyl, CONH2 or SO2NR 4 R 5 wherein R 4 and R 5 together with the nitrogen atom to which they are attached form an azetidine ring, a pyrrolidine ring or a piperidine ring); or R 1 is [Chemical formula] <00004​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​In another embodiment, compounds represented by formula (I) or pharmaceutically acceptable salts of compounds represented by formula (I) are provided (where the stereochemistry is undefined and is, for example, a racemic mixture or a mixture of diastereomers).

[0021] In another embodiment, a pharmaceutical formulation is provided comprising a compound represented by formula (I) or a pharmaceutically acceptable salt of a compound represented by formula (I), and a pharmaceutically acceptable diluent, excipient, and / or inert carrier.

[0022] In further embodiments, a pharmaceutical formulation for treating a condition in which inhibition of dipeptidyl peptidase 1 (DPP1) is beneficial is provided, comprising a compound represented by formula (I) or a pharmaceutically acceptable salt of a compound represented by formula (I).

[0023] In further embodiments, compounds represented by formula (I) or pharmaceutically acceptable salts of compounds represented by formula (I) are provided for the therapeutic (particularly for prophylactic or treatment) of respiratory diseases in mammals (particularly humans).

[0024] In further embodiments, compounds represented by formula (I) or pharmaceutically acceptable salts of compounds represented by formula (I) are provided for the therapeutic (particularly for prophylactic or treatment) of asthma in mammals (particularly humans).

[0025] In further embodiments, compounds represented by formula (I) or pharmaceutically acceptable salts of compounds represented by formula (I) are provided for the therapeutic (particularly prophylactic or treatment) of COPD in mammals (particularly humans).

[0026] Further embodiments provide the use of a compound represented by formula (I) or a pharmaceutically acceptable salt of a compound represented by formula (I) for the manufacture of pharmaceuticals for the treatment and prevention of respiratory diseases.

[0027] Further embodiments provide the use of a compound represented by formula (I) or a pharmaceutically acceptable salt of a compound represented by formula (I) for the manufacture of medicines for the treatment and prevention of asthma.

[0028] Further embodiments provide the use of a compound represented by formula (I) or a pharmaceutically acceptable salt of a compound represented by formula (I) for the manufacture of pharmaceuticals for the treatment and prevention of COPD.

[0029] In a further embodiment, administration of a compound represented by formula (I) or a pharmaceutically acceptable salt of a compound represented by formula (I) causes a decrease in DPP1 levels in mammals (particularly humans).

[0030] In further embodiments, administration of the compound represented by formula (I) or a pharmaceutically acceptable salt of the compound represented by formula (I) causes a decrease in the levels of DPP1, neutrophil elastase, cathepsin G, and proteinase 3 in mammals (particularly humans).

[0031] In a further embodiment, administration of the compound represented by formula (I) or a pharmaceutically acceptable salt of the compound represented by formula (I) causes a decrease in DPP1 activity in mammals (particularly humans).

[0032] In a further embodiment, administration of the compound represented by formula (I) or a pharmaceutically acceptable salt of the compound represented by formula (I) causes a decrease in DPP1 activity, neutrophil elastase activity, cathepsin G activity, and proteinase 3 activity in mammals (particularly humans).

[0033] In another embodiment, a method for producing a compound represented by formula (I) or a pharmaceutically acceptable salt of a compound represented by formula (I), and intermediates used in the production method are provided.

[0034] According to another aspect, formula (XXIV): [ka] [In the formula, R 8 C 1-4 Selected from alkyl or aryl (where the aryl is R 1 It may be replaced by; R 9 and R 10 This represents a 5- to 7-membered saturated or unsaturated ring which, together with the nitrogen atom to which they are bonded, may contain one other heteroatom which is oxygen, nitrogen, or sulfur (where the ring may be fused with a (C3-C8) cycloalkyl, heterocycloalkyl, aryl, or heteroaryl ring); or R 9 and R 10 This represents a 6- to 10-membered bridged bicyclic ring, which may be condensed with a (C3-C8) cycloalkyl, heterocycloalkyl, aryl, or heteroaryl ring together with the nitrogen atoms to which they are bonded. Compounds represented by or pharmaceutically acceptable salts thereof are provided.

[0035] Further embodiments provide compounds represented by formula (XXIV) or pharmaceutically acceptable salts of compounds represented by formula (XXIV) for the treatment (particularly for prevention or treatment) of respiratory diseases in mammals (particularly humans).

[0036] The compound represented by formula (I) as illustrated herein exhibits IC12 for DPP1 in an enzyme activity assay (e.g., Test A1 or Test A2 below). 50 The concentration is less than 100 nmol / L. The compound represented by formula (I) also exhibits a promising pharmacological profile by differentiating between desirable and undesirable effects in vivo. [Brief explanation of the drawing]

[0037] [Figure 1]Figure 1 shows the powder X-ray diffraction pattern of Example 2: (2S)-N-{(1S)-1-cyano-2-[4-(3-methyl-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl)phenyl]ethyl}-1,4-oxazepan-2-carboxamide, form A. [Figure 2] Figure 2 shows the powder X-ray diffraction pattern of Example 2: (2S)-N-{(1S)-1-cyano-2-[4-(3-methyl-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl)phenyl]ethyl}-1,4-oxazepan-2-carboxamide, form B. [Figure 3] Figure 3 shows the powder X-ray diffraction pattern of Example 2: (2S)-N-{(1S)-1-cyano-2-[4-(3-methyl-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl)phenyl]ethyl}-1,4-oxazepan-2-carboxamide, form C. [Figure 4] Figure 4 shows the powder X-ray diffraction pattern of Example 2: (2S)-N-{(1S)-1-cyano-2-[4-(3-methyl-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl)phenyl]ethyl}-1,4-oxazepan-2-carboxamide xinafoate, form A. [Figure 5] Figure 5 shows the powder X-ray diffraction pattern of Example 2: (2S)-N-{(1S)-1-cyano-2-[4-(3-methyl-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl)phenyl]ethyl}-1,4-oxazepan-2-carboxamide R-mandelate, form A. [Modes for carrying out the invention]

[0038] Detailed explanation This detailed description is intended to inform those skilled in the art of the present invention, its spirit, and its practical applications, so that they may readily utilize it. This description and its specific examples illustrate embodiments of the present invention, but are for illustrative purposes only. Therefore, the present invention is not limited to the exemplary embodiments described herein. In addition, for clarity, various features of the present invention described in relation to separate embodiments may be combined to form a single embodiment. Conversely, for brevity, various features of the present invention described in a single embodiment may be combined to form a subcombination thereof.

[0039] The following are definitions of various terms used in this specification and in the claims to describe the present invention.

[0040] To avoid misunderstanding, wherever a base is conditioned in this specification by the phrase "as defined above," this base naturally encompasses all of the broadest definition initially provided and all other definitions of that base.

[0041] To avoid misunderstanding, it goes without saying that "C" is used in this specification. 1-3 "A carbon group" refers to a carbon group having one, two, or three carbon atoms.

[0042] In this specification, unless otherwise specified, the term "alkyl" includes both linear and branched alkyl groups and may be, but is not limited to, methyl, ethyl, n-propyl, or i-propyl.

[0043] In this specification, unless otherwise specified, the term “medically acceptable” is used to characterize a moiety (e.g., a salt, dosage form, or excipient) as suitable for use in accordance with sound medical judgment. Generally, a medically acceptable moiety has one or more benefits that outweigh any adverse effects it may have. Adverse effects include, for example, excessive toxicity, irritation, allergic reactions, and other problems and complications.

[0044] R 1 ~R 7 A compound represented by formula (I) is disclosed, wherein X, Y, and Q are as defined in formula (I).

[0045] In one embodiment, R 1 teeth, [ka] and; R 2 These are hydrogen, F, Cl, Br, and OSO2C. 1-3 Alkyl or C 1-3 Selected from alkyl groups; R 3 These are hydrogen, F, Cl, Br, CN, CF3, SO2C 1-3 Alkyl, CONH2, or SO2NR 4 R 5 Selected from, here, R 4 and R 5 These, together with the nitrogen atom to which they are bonded, form an azetidine ring, a pyrrolidine ring, or a piperidine ring.

[0046] In a further embodiment, R 1 teeth, [ka] and; R 2 is hydrogen, F, Cl or C 1-3 Selected from alkyl groups; R 3 These are hydrogen, F, Cl, CN, or SO2C. 1-3 Selected from alkyl groups.

[0047] In a further embodiment, R 1 teeth, [ka] and; R 2 is hydrogen, F or C1-3 Selected from alkyl groups; R 3 This is selected from hydrogen, F, or CN.

[0048] In a further embodiment, R 1 teeth, [ka] Selected from; X is selected from O, S, or CF2; Y is selected from O or S; Q is selected from CH or N; R 6 C 1-3 Selected from alkyl (where C 1-3 The alkyl group may be substituted with 1, 2, or 3 F atoms, and also OH,OC 1-3 Alkyl, N(C 1-3 It may be substituted with one substituent selected from alkyl)2, cyclopropyl, or tetrahydropyran); R 7 This is selected from hydrogen, F, Cl, or CH3.

[0049] In a further embodiment, R 1 teeth, [ka] Selected from; X is selected from O, S, or CF2; Y is selected from O or S; R 6 C 1-3 Selected from alkyl (where C 1-3 The alkyl group may be substituted with 1, 2, or 3 F atoms, and also OH,OC 1-3 Alkyl, N(C 1-3 It may be substituted with one substituent selected from alkyl)2, cyclopropyl, or tetrahydropyran); R7 This is selected from hydrogen, F, Cl, or CH3.

[0050] In a further embodiment, R 1 teeth, [ka] Selected from; X is selected from O, S, or CF2; R 6 C 1-3 Selected from alkyl (where C 1-3 The alkyl group may be substituted with 1, 2, or 3 F atoms. R 7 This is selected from hydrogen, F, Cl, or CH3.

[0051] In a further embodiment, R 1 teeth, [ka] Selected from; X is O; R 6 C 1-3 Selected from alkyl (where C 1-3 The alkyl group may be substituted with 1, 2, or 3 F atoms; R 7 It is hydrogen.

[0052] In one embodiment, R 2 These are hydrogen, F, Cl, Br, and OSO2C. 1-3 Alkyl or C 1-3 Selected from alkyl groups.

[0053] In a further embodiment, R 2 is hydrogen, F, Cl or C 1-3 Selected from alkyl groups.

[0054] In a further embodiment, R 2 is hydrogen, F or C 1-3It is selected from alkyl.

[0055] In one embodiment, R 3 is hydrogen, F, Cl, Br, CN, CF3, SO2C 1-3 alkyl, CONH2 or SO2NR 4 R 5 is selected from, where R 4 and R 5 together with the nitrogen atom to which they are attached form an azetidine ring, a pyrrolidine ring or a piperidine ring.

[0056] In a further embodiment, R 3 is selected from hydrogen, F, Cl, CN or SO2C 1-3 alkyl.

[0057] In an even further embodiment, R 3 is selected from hydrogen, F or CN.

[0058] In one embodiment, R 6 is selected from C 1-3 alkyl, where the C 1-3 alkyl may be substituted by 1, 2 or 3 Fs and may also be substituted by one substituent selected from OH, OC 1-3 alkyl, N(C 1-3 alkyl)2, cyclopropyl or tetrahydropyran.

[0059] In a further embodiment, R 6 is selected from C 1-3 alkyl, where the C 1-3 alkyl may be substituted by 1, 2 or 3 Fs.

[0060] In an even further embodiment, R 6 is selected from methyl and ethyl.

[0061] In an even further embodiment, R 6 is methyl.

[0062] In one embodiment, R 7 This is selected from hydrogen, F, Cl, or CH3.

[0063] In a further embodiment, R 7 It is hydrogen.

[0064] Combining one or more of the above embodiments provides further specific embodiments of the present invention.

[0065] In one embodiment, the compound represented by formula (I) is selected from the following: (2S)-N-[(1S)-1-cyano-2-(4'-cyanobiphenyl-4-yl)ethyl]-1,4-oxazepan-2-carboxamide, (2S)-N-{(1S)-1-cyano-2-[4-(3-methyl-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl)phenyl]ethyl}-1,4-oxazepan-2-carboxamide, (2S)-N-{(1S)-1-cyano-2-[4-(3,7-dimethyl-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl)phenyl]ethyl}-1,4-oxazepan-2-carboxamide, 4'-[(2S)-2-cyano-2-{[(2S)-1,4-oxazepan-2-ylcarbonyl]amino}ethyl]biphenyl-3-ylmethanesulfonate, (2S)-N-{(1S)-1-cyano-2-[4-(3-methyl-1,2-benzoxazol-5-yl)phenyl]ethyl}-1,4-oxazepan-2-carboxamide, (2S)-N-{(1S)-1-cyano-2-[4'-(trifluoromethyl)biphenyl-4-yl]ethyl}-1,4-oxazepan-2-carboxamide, (2S)-N-[(1S)-1-cyano-2-(3',4'-difluorobiphenyl-4-yl)ethyl]-1,4-oxazepan-2-carboxamide, (2S)-N-{(1S)-1-cyano-2-[4-(6-cyanopyridine-3-yl)phenyl]ethyl}-1,4-oxazepan-2-carboxamide, (2S)-N-{(1S)-1-cyano-2-[4-(4-methyl-3-oxo-3,4-dihydro-2H-1,4-benzothiadin-6-yl)phenyl]ethyl}-1,4-oxazepan-2-carboxamide, (2S)-N-{(1S)-1-cyano-2-[4-(3-ethyl-7-methyl-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl)phenyl]ethyl}1,4-oxazepan-2-carboxamide, (2S)-N-[(1S)-1-cyano-2-{4-[3-(2-hydroxy-2-methylpropyl)-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl]phenyl}ethyl]-1,4-oxazepan-2-carboxamide, (2S)-N-[(1S)-1-cyano-2-{4-[3-(2,2-difluoroethyl)-7-fluoro-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl]phenyl}ethyl]-1,4-oxazepan-2-carboxamide, (2S)-N-[(1S)-1-cyano-2-(4-{3-[2-(dimethylamino)ethyl]-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl}phenyl)ethyl]-1,4-oxazepan-2-carboxamide, (2S)-N-{(1S)-1-cyano-2-[4-(3,3-difluoro-1-methyl-2-oxo-2,3-dihydro-1H-indole-6-yl)phenyl]ethyl}-1,4-oxazepan-2-carboxamide, (2S)-N-{(1S)-1-cyano-2-[4-(7-fluoro-3-methyl-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl)phenyl]ethyl}-1,4-oxazepan-2-carboxamide, (2S)-N-{(1S)-1-cyano-2-[4-(3-ethyl-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl)phenyl]ethyl}1,4-oxazepan-2-carboxamide, (2S)-N-[(1S)-1-cyano-2-{4-[3-(cyclopropylmethyl)-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl]phenyl}ethyl]-1,4-oxazepan-2-carboxamide, (2S)-N-[(1S)-1-cyano-2-{4-[3-(2-methoxyethyl)-2-oxo-2,3-dihydro-1,3-benzothiazole-5-yl]phenyl}ethyl]-1,4-oxazepan-2-carboxamide,

[0066] (2S)-N-[(1S)-1-cyano-2-{4-[2-oxo-3-(propan-2-yl)-2,3-dihydro-1,3-benzoxazole-5-yl]phenyl}ethyl]-1,4-oxazepan-2-carboxamide, (2S)-N-{(1S)-1-cyano-2-[4-(4-methyl-3-oxo-3,4-dihydro-2H-1,4-benzoxazine-6-yl)phenyl]ethyl}-1,4-oxazepan-2-carboxamide, (2S)-N-[(1S)-1-cyano-2-{4-[3-(2-methoxyethyl)-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl]phenyl}ethyl]-1,4-oxazepan-2-carboxamide, (2S)-N-{(1S)-1-cyano-2-[4-(5-cyanothiophen-2-yl)phenyl]ethyl}-1,4-oxazepan-2-carboxamide, (2S)-N-[(1S)-2-(4'-carbamoyl-3'-fluorobiphenyl-4-yl)-1-cyanoethyl]-1,4-oxazepan-2-carboxamide, (2S)-N-{(1S)-1-cyano-2-[4-(1-methyl-2-oxo-1,2-dihydroquinoline-7-yl)phenyl]ethyl}-1,4-oxazepan-2-carboxamide, (2S)-N-[(1S)-1-cyano-2-{4-[2-oxo-3-(tetrahydro-2H-pyran-4-ylmethyl)-2,3-dihydro-1,3-benzoxazole-5-yl]phenyl}ethyl]-1,4-oxazepan-2-carboxamide, (2S)-N-{(1S)-2-[4-(7-chloro-3-methyl-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl)phenyl]-1-cyanoethyl}-1,4-oxazepan-2-carboxamide, (2S)-N-[(1S)-1-cyano-2-{4-[3-(2,2-difluoroethyl)-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl]phenyl}ethyl]-1,4-oxazepan-2-carboxamide, (2S)-N-[(1S)-1-cyano-2-{4-[2-oxo-3-(2,2,2-trifluoroethyl)-2,3-dihydro-1,3-benzoxazole-5-yl]phenyl}ethyl]-1,4-oxazepan-2-carboxamide, (2S)-N-{(1S)-1-cyano-2-[4-(3-methyl-2-oxo-2,3-dihydro-1,3-benzothiazole-5-yl)phenyl]ethyl}-1,4-oxazepan-2-carboxamide, (2S)-N-{(1S)-1-cyano-2-[4'-(methylsulfonyl)biphenyl-4-yl]ethyl}-1,4-oxazepan-2-carboxamide, (2S)-N-{(1S)-2-[4'-(azetidine-1-ylsulfonyl)biphenyl-4-yl]-1-cyanoethyl}-1,4-oxazepan-2-carboxamide, (2S)-N-[(1S)-1-cyano-2-(4'-fluorobiphenyl-4-yl)ethyl]-1,4-oxazepan-2-carboxamide, (2S)-N-{(1S)-2-[4-(1,3-benzothiazole-5-yl)phenyl]-1-cyanoethyl}-1,4-oxazepan-2-carboxamide, (2S)-N-[(1S)-1-cyano-2-(4'-cyanobiphenyl-4-yl)ethyl]-1,4-oxazepan-2-carboxamide, or (2S)-N-{(1S)-1-cyano-2-[4-(4-methyl-3-oxo-1,2,3,4-tetrahydroquinoxaline-6-yl)phenyl]ethyl}-1,4-oxazepan-2-carboxamide, and its medicinally acceptable salts.

[0067] It should be noted that any one of these specific compounds may be disclaimed from the embodiments of the present invention described herein.

[0068] Another embodiment is a product obtained by any of the manufacturing methods or examples described herein.

[0069] Pharmacological properties The compounds represented by formula (I) and their pharmaceutically acceptable salts are active as pharmaceutical formulations, particularly as inhibitors of dipeptidyl peptidase 1 activity, and thus can be used to treat obstructive airway diseases, including: bronchial asthma of all severity, both intermittent and persistent, allergic asthma, endogenous asthma, exogenous asthma, exercise-induced asthma, drug-induced (aspirin-induced and NSAID-induced) asthma and dust-induced asthma, as well as other causes of airway hyperresponsiveness; chronic obstructive pulmonary disease (COPD); bronchitis, including infectious bronchitis and eosinophilic bronchitis; emphysema; bronchiectasis; cystic fibrosis; sarcoidosis; α1-antitrypsin deficiency; farmer's lung and related diseases; hypersensitivity pneumonitis; idiopathic fibrotic alveolitis, idiopathic interstitial pneumonia, and antineoplastic syndrome. Pulmonary fibrosis, including fibrosis associated with drug therapy and chronic infections (including tuberculosis and aspergillosis) and other fungal infections; complications of lung transplantation; vasculitic and thrombotic disorders of the pulmonary vascular structure, and pulmonary hypertension; chronic cough associated with inflammatory and secretory conditions of the airways, and treatment of iatrogenic cough, including antitussive activity; acute and chronic rhinitis, including drug-induced rhinitis and vasomotor rhinitis; perennial and seasonal allergic rhinitis, including neurogenic rhinitis (hay fever); nasal polyposis; the common cold, and acute viral infections, including infections caused by RSV (respiratory polynucleus) virus, influenza, coronavirus (including SARS), and adenovirus; acute lung injury; acute respiratory distress syndrome (ARDS); and exacerbations of each of the above inspiratory disease conditions, particularly exacerbations of any type of asthma or COPD.

[0070] Thus, therapeutic compounds represented by formula (I) defined above, or pharmaceutically acceptable salts thereof, are provided.

[0071] In a further embodiment, the use of the compound represented by formula (I) defined above or a pharmaceutically acceptable salt thereof in the manufacture of a therapeutic pharmaceutical is provided.

[0072] In the context of this specification, the term “treatment” also includes “prevention” unless otherwise specifically indicated. The terms “therapeutic” and “therapeutically” shall also be interpreted accordingly.

[0073] Prevention is thought to be particularly related to the treatment of individuals who have suffered a past episode of the disease or condition in question, or who are considered to be at high risk of developing the disease or condition in question. Individuals at risk of developing a particular disease or condition generally include those with a family history of the disease or condition, or those identified as particularly susceptible to the disease or condition through genetic testing or screening.

[0074] In particular, the compounds of the present invention (including pharmaceutically acceptable salts) can be used in the treatment of asthma {e.g., bronchial asthma, allergic asthma, endogenous asthma, exogenous asthma or dust asthma, especially chronic or refractory asthma (e.g., delayed-onset asthma or airway hyperresponsiveness)}, chronic obstructive pulmonary disease (COPD), or allergic rhinitis.

[0075] Also provided are methods for treating or reducing the risk of obstructive airway diseases or conditions (e.g., asthma or COPD), comprising administering a therapeutically effective amount of a compound represented by formula (I) as defined above or a pharmaceutically acceptable salt thereof to a patient in need of such treatment or reduction.

[0076] In a further embodiment, the use of the compound represented by formula (I) defined above or a pharmaceutically acceptable salt thereof in the manufacture of a pharmaceutical for the treatment of COPD is provided.

[0077] In a further embodiment, the use of the compound represented by formula (I) as defined above or a pharmaceutically acceptable salt thereof in the manufacture of a pharmaceutical for the treatment of asthma is provided.

[0078] In a further embodiment, the use of the compound represented by formula (I) as defined above or a pharmaceutically acceptable salt thereof in the manufacture of a pharmaceutical for the treatment of allergic rhinitis is provided.

[0079] In a further embodiment, compounds represented by formula (I) defined above or pharmaceutically acceptable salts thereof are provided for the treatment of allergic rhinitis.

[0080] In a further embodiment, compounds represented by formula (I) defined above or pharmaceutically acceptable salts thereof are provided for the treatment of COPD.

[0081] In a further embodiment, compounds represented by formula (I) defined above or pharmaceutically acceptable salts thereof are provided for the treatment of asthma.

[0082] Combination therapy The compound represented by formula (I) or a pharmaceutically acceptable salt thereof may be administered in combination with another compound used to treat the above condition.

[0083] The present invention also relates to combination therapies in which a compound of the present invention or a pharmaceutically acceptable salt thereof and a second active ingredient are administered simultaneously, sequentially, or in combination for the treatment of one or more of the above conditions. Such combinations may be used in combination with one or more further active ingredients.

[0084] The present invention further comprises a compound of the present invention or a pharmaceutically acceptable salt thereof, and a glucocorticoid receptor agonist (steroidal or non-steroidal), such as triamcinolone, triamcinolone acetonide, prednisone, mometasone furoate, lotebredonol etavonate, fluticasone propionate, fluocinolone acetonide, dexamethasone cypesylate, desisobutyryl ciclesonide, clobetasol propionate, ciclesonide, butixocort propionate, budesonide, and dipropionate. This relates to combinations with clometasone, alclometasone dipropionate, 2,2,2-trifluoro-N-[(1S,2R)-2-[1-(4-fluorophenyl)indazole-5-yl]oxy-2-(3-methoxyphenyl)-1-methyl-ethyl]acetamide, or 3-[5-[(1R,2S)-2-(2,2-difluoropropanoylamino)-1-(2,3-dihydro-1,4-benzodioxin-6-yl)propoxy]indazole-1-yl]-N-[(3R)-tetrahydrofuran-3-yl]benzamide.

[0085] The present invention further includes a compound of the present invention or a pharmaceutically acceptable salt thereof, and a p38 antagonist, for example, PH797804 (3-[3-bromo-4-(2,4-difluorobenzyloxy)-6-methyl-2-oxo-2H-pyridine-1-yl]-4,N-dimethylbenzamide), rosmapimod, PF03715455 (1-[5-tert-butyl-2-(3-chloro-4-hydroxyphenyl)pyrazole-3 The combination with -yl]-3-[[2-[[3-[2-(2-hydroxyethylsulfanyl)phenyl]-[1,2,4]triazolo[4,3-a]pyridine-6-yl]sulfanyl]phenyl]methyl]urea) or N-cyclopropyl-3-fluoro-4-methyl-5-[3-[[1-[2-[2-(methylamino)ethoxy]phenyl]cyclopropyl]amino]-2-oxopyrazine-1-yl]benzamide.

[0086] The present invention further relates to the compounds of the present invention or pharmaceutically acceptable salts thereof, and phosphodiesterase (PDE) inhibitors, such as methylxanthanine (including theophylline and aminophylline), or selective PDE isozyme inhibitors (including PDE4 inhibitors or inhibitors of isoform PDE4D), such as tetomirast, roflumilast, ogremilast, ibudilast, GPD-1116 (3-benzyl-5-phenyl-1H-pyrazolo[4,3-c][1,8]naphthyridine-4-one), ronomilast, and NVP ABE. This relates to combinations with 171 (4-[8-(2,1,3-benzoxadiazole-5-yl)-1,7-naphthyridine-6-yl]benzoic acid), RPL554 (2-[(2E)-9,10-dimethoxy-4-oxo-2-(2,4,6-trimethylphenyl)imino-6,7-dihydropyrimide[6,1-a]isoquinoline-3-yl]ethylurea), CHF5480 ([(Z)-2-(3,5-dichloro-4-pyridyl)-1-(3,4-dimethoxyphenyl)vinyl](2S)-2-(4-isobutylphenyl)propanoate), or GSK256066 (6-[3-(dimethylcarbamoyl)phenyl]sulfonyl-4-(3-methoxyanilino)-8-methylquinoline-3-carboxamide).

[0087] The present invention further relates to combinations of the compound of the present invention or a pharmaceutically acceptable salt thereof with a chemokine receptor function modifier, for example, an antagonist of CCR1, CCR2, CCR2A, CCR2B, CCR3, CCR4, CCR5, CCR6, CCR7, CCR8, CCR9, CCR10 or CCR11 (for the CC family), for example, a CCR1, CCR2B or CCR5 receptor antagonist; CXCR1, CXCR2, CXCR3, CXCR4 or CXCR5 (for the CXC family), for example, a CXCR2 or CXCR3 receptor antagonist; or CX3CR1 for the C-X3-C family. For example, the present invention includes the compounds of the present invention and PS-031291 (pyrrolidin-1,2-dicarboxylic acid 2-[(4-chlorobenzyl)-methyl-amide]1-[(4-trifluoromethylphenyl)-amide]), CCX-354 (1-[4-(4-chloro-3-methoxyphenyl)piperazine-1-yl]-2-[3-(1H-imidazole-2-yl)pyrazolo[3,4-b]pyridine-1-yl]ethanone), vicribiroc, maraviroc, seniclibiroc, navalixin (2-hydroxy-N,N-dimethyl-3-[[2-[[(1R)-1-(5-methyl-2-furyl)propyl]amino]-3,4-dioxocyclobuten-1-yl]amino]benzamide), SB656933 (1-(2-chloro-3-fluorophenyl)-3-( This relates to combinations with 4-chloro-2-hydroxy-3-piperazine-1-ylsulfonylphenyl)urea), N-[2-[(2,3-difluorophenyl)methylsulfanyl]-6-[(1R,2S)-2,3-dihydroxy-1-methyl-propoxy]pyrimidine-4-yl]azetidine-1-sulfonamide, N-[6-[(1R,2S)-2,3-dihydroxy-1-methyl-propoxy]-2-[(4-fluorophenyl)methylsulfanyl]pyrimidine-4-yl]-3-methylazetidine-1-sulfonamide, or N-[2-[(2,3-difluorophenyl)methylsulfanyl]-6-[[(1R,2R)-2,3-dihydroxy-1-methyl-propyl]amino]pyrimidine-4-yl]azetidine-1-sulfonamide.

[0088] The present invention further relates to compounds of the present invention or pharmaceutically acceptable salts thereof, and leukotriene biosynthesis inhibitors, 5-lipoxygenase (5-LO) inhibitors, or 5-lipoxygenase-activating protein (FLAP) antagonists, such as TA270 (4-hydroxy-1-methyl-3-octyloxy-7-sinapinoylamino-2(1H)-quinolinone), PF-4191834 (2H-pyran-4-carb Xamide, Tetrahydro-4-[3-[[4-(1-methyl-1H-pyrazole-5-yl)phenyl]thio]phenyl]-), Setileuton, CMI977(1-[4-[(2S,5S)-5-[(4-fluorophenoxy)methyl]tetrahydrofuran-2-yl]buta-3-inyl]-1-hydroxyurea), Fiboflapon(3-[3-tert-butylsulfanyl-1-[[4-(6-ethoxy -3-pyridyl)phenyl]methyl]-5-[(5-methyl-2-pyridyl)methoxy]indole-2-yl]-2,2-dimethyl-propanoic acid), GSK2190915 (1H-indole-2-propanoic acid, 3-[(1,1-dimethylethyl)thio]-1-[[4-(6-methoxy-3-pyridinyl)phenyl]methyl]-α,α-dimethyl-5-[(2-pyridinyl)methoxy]-), lycopherone, quiflapon (3-[3-t This relates to combinations with ert-butylsulfanyl-1-[(4-chlorophenyl)methyl]-5-(2-quinolylmethoxy)indole-2-yl]-2,2-dimethylpropanoic acid, beriflapon ((2R)-2-cyclopentyl-2-[4-(2-quinolylmethoxy)phenyl]acetic acid), ABT080 (4,4-bis[4-(2-quinolylmethoxy)phenyl]pentanoic acid), diloton, zafirlukast, or montelukast.

[0089] The present invention further comprises a compound of the present invention or a pharmaceutically acceptable salt thereof, and a CRTh2 antagonist or DP2 antagonist, for example, ACT129968 (2-[2-[(5-acetyl-2-methoxyphenyl)methylsulfanyl]-5-fluoro-benzimidazole-1-yl]acetic acid), AMG853 (2-[4-[4-(tert-butylcarbamoyl)-2-[(2-chloro-4-cyclopropyl-phenyl)sulfonylamino]phenoxy]-5-chloro-2-fluorophenyl]acetic acid), AM211 (2-[3-[2-[[benzylcarbamoyl(ethyl)amino]methyl] This relates to combinations with (2S)-2-[4-(trifluoromethyl)phenyl]-4-methoxyphenyl]acetic acid, 2-[4-acetamido-3-(4-chlorophenyl)sulfanyl-2-methylindole-1-yl]acetic acid, (2S)-2-[4-chloro-2-(2-chloro-4-ethylsulfonylphenoxy)phenoxy]propanoic acid, 2-[4-chloro-2-[2-fluoro-4-(4-fluorophenyl)sulfonylphenyl]phenoxy]acetic acid, or (2S)-2-[2-[3-chloro-4-(2,2-dimethylpyrrolidine-1-carbonyl)phenyl]-4-fluorophenoxy]propanoic acid.

[0090] The present invention further relates to combinations of the compound of the present invention or a pharmaceutically acceptable salt thereof with a myeloperoxidase antagonist, such as resveratrol, picetanol, or 1-(2-isopropoxyethyl)-2-thioxo-5H-pyrrolo[3,2-d]pyrimidine-4-one.

[0091] In a further embodiment of the present invention, a compound of the present invention or a pharmaceutically acceptable salt thereof, as defined above, a) Toll-like receptor agonists (e.g., TLR7 agonists or TLR9 agonists); b) Adenosine antagonists; c) Glucocorticoid receptor agonists (steroidal or non-steroidal); d) p38 Antagonist; e) PDE4 antagonist; f) Chemokine receptor function modifiers (e.g., CCR1 receptor antagonists, CCR2B receptor antagonists, CCR5 receptor antagonists, CXCR2 receptor antagonists, or CXCR3 receptor antagonists); or g) CRTh2 antagonist A pharmaceutical composition is provided, comprising at least one active ingredient selected from (for use as a pharmacopoeia for the treatment of one of the diseases or conditions described herein, such as COPD, asthma, or allergic rhinitis).

[0092] In one embodiment, the compound of the present invention or a pharmaceutically acceptable salt thereof is administered simultaneously or sequentially with one or more further active ingredients selected from those defined above. For example, the compound of the present invention or a pharmaceutically acceptable salt thereof may be administered simultaneously or sequentially with a further pharmaceutical composition for use as a pharmacopoeia for the treatment of one of the above diseases or conditions, such as a respiratory condition (e.g., COPD, asthma, or allergic rhinitis). This further pharmaceutical composition may be a pharmacopoeia that the patient may already be prescribed (e.g., an existing standard care medication) and may itself be a composition comprising one or more further active ingredients selected from those defined above.

[0093] Pharmaceutical composition For the therapeutic uses described above, the dosage will vary depending on the compound used, the mode of administration, the desired treatment, and the disease being treated. For example, the daily dose of the compound of the present invention may range from 0.05 μg (μg / kg) to 100 μg (μg / kg) per kg of body weight when inhaled. Alternatively, when the compound is administered orally, the daily dose of the compound of the present invention may range from 0.01 μg (μg / kg) to 100 mg (mg / kg) per kg of body weight.

[0094] The compound represented by formula (I) or a pharmaceutically acceptable salt thereof may be used alone, but is generally administered in the form of a pharmaceutical composition in which the formula (I) compound / salt (active ingredient) is accompanied by a pharmaceutically acceptable adjuvant, diluent, or carrier. Conventional procedures for the selection and preparation of suitable pharmaceutical formulations are described, for example, in “Pharmaceuticals - The Science of Dosage Form Designs”, ME Aulton, Churchill Livingstone, 2nd Ed. 2002.

[0095] Depending on the mode of administration, the pharmaceutical composition preferably contains the active ingredient in an amount of 0.05 to 99% w (weight %), more preferably 0.05 to 80% w, even more preferably 0.10 to 70% w, and still more preferably 0.10 to 50% w (all based on weight %) of the entire composition.

[0096] The present invention also provides pharmaceutical compositions comprising a compound represented by formula (I) as defined above or a pharmaceutically acceptable salt thereof, accompanied by a pharmaceutically acceptable adjuvant, diluent, or carrier.

[0097] The present invention further provides a method for preparing a pharmaceutical composition of the present invention, comprising mixing a compound represented by formula (I) defined above or a pharmaceutically acceptable salt thereof with a pharmaceutically acceptable adjuvant, diluent, or carrier.

[0098] The pharmaceutical composition may be administered topically (e.g., to the skin, or lungs and / or airways) in dosage forms such as creams, liquids, suspensions, heptafluoroalkane (HFA) aerosols, and dry powders (e.g., formulations in inhalers known as Turbuhaler®); systemically by oral administration in dosage forms such as tablets, capsules, syrups, powders, or granules; parenterally by sterile liquids, suspensions, or emulsions for injection (including intravenous, subcutaneous, intramuscular, intravascular, or infusion); or rectally by suppositories.

[0099] For oral administration, the compounds of the present invention may be mixed with an adjuvant, diluent or carrier, such as lactose, saccharose, sorbitol, or mannitol; starch, such as potato starch, corn starch, or amylopectin; cellulose derivative; binder, such as gelatin or polyvinylpyrrolidone; disintegrant, such as a cellulose derivative; and / or lubricant, such as magnesium stearate, calcium stearate, polyethylene glycol, wax, or paraffin, and then compressed into tablets. If coated tablets are required, the core prepared as described above may be coated with a suitable polymer dissolved or dispersed in water or an easily volatile organic solvent. Alternatively, the tablets may be coated with a concentrated sugar solution that may contain, for example, gum arabic, gelatin, talc, and titanium dioxide.

[0100] For the manufacture of soft gelatin capsules, the compound of the present invention may be mixed with, for example, vegetable oil or polyethylene glycol. Hard gelatin capsules may contain granules of the compound using pharmaceutically acceptable excipients such as those described above for tablets. Liquid or semi-solid formulations of the compound of the present invention may also be filled into hard gelatin capsules.

[0101] Liquid formulations for oral administration may be in the form of syrups, liquids, or suspensions. Liquid formulations may, for example, contain the compounds of the present invention, balance-being sugar, and a mixture of ethanol, water, glycerol, and propylene glycol. Optionally, such liquid formulations may contain colorants, flavorings, saccharin, and / or carboxymethylcellulose as thickening agents. Furthermore, other additives known to those skilled in the art may be used when preparing oral formulations.

[0102] Manufacturing of compounds The present invention further provides a method for producing the compound represented by formula (I) as defined above.

[0103] General manufacturing methods Those skilled in the art will recognize that the compounds of the present invention can be produced in various ways using known methods. The following routes simply describe some of the methods that can be used to synthesize the compound represented by formula (I).

[0104] The present invention further relates to a method for producing a compound represented by formula (I) as defined above or a pharmaceutically acceptable salt thereof, wherein formula (II): [ka] [In the formula, R 1 This is as defined in equation (I). The compound shown by formula (III): [ka] [In the formula, PG represents a protecting group (e.g., tert-butoxycarbonyl)] This involves reacting with the compound shown, and optionally, then proceeding to the following steps: ● Converting a compound represented by formula (I) to another compound represented by formula (I), ● Remove the protecting group. ● To form a pharmaceutically acceptable salt. The present invention provides a method which may include performing one or more of the following:

[0105] The method is preferably carried out in the presence of a base such as DiPEA or TEA, and one or more activators such as EDCI, 2-pyridinol-1-oxide, or T3P. The reaction is preferably carried out at a temperature in the range of 20°C to 100°C, particularly at ambient temperature (25°C), in the presence of an organic solvent such as DMF or DCM.

[0106] The compound represented by formula (II) is represented by formula (IV): [ka] [In the formula, PG represents a protecting group (e.g., tert-butoxycarbonyl)] The compound shown can be produced by reacting it with a suitable reagent to remove the protecting group PG. An example of a suitable reagent is formic acid.

[0107] The compound represented by formula (IV) is formed in the presence of a catalyst such as Pd(dppf)Cl2·DCM or 1,1-bis(di-tert-butylphosphin)ferrocenepalladium dichloride and a base such as potassium carbonate or sodium carbonate, as shown in formula (V): [ka] [In the formula, PG represents a protecting group (e.g., tert-butoxycarbonyl), and Hal represents a halogen (e.g., I or Br)] The compound shown by formula (VI): [ka] [In the formula, R 1 This is as defined in equation (I). It can be produced by reacting it with the compound shown or its ester. The reaction is conveniently carried out in a solvent such as a dioxane / water mixture or an ACN / water mixture at a temperature in the range of 20°C to 100°C, particularly at 75°C.

[0108] The compound represented by formula (V) is represented by formula (VII): [ka] [In the formula, PG represents a protecting group (e.g., tert-butoxycarbonyl), and Hal represents a halogen (e.g., I or Br)] The compounds shown can be prepared using standard literature-described procedures for amide dehydration, for example, at temperatures ranging from -20°C to 100°C, for example at 0°C, in a solvent such as DCM or DMF, with or without a base such as DiPEA, and using a reagent such as Burgess reagent or T3P.

[0109] The compound represented by formula (VII) is formed using the standard literature-described procedure for amide formation, for example, in the presence of a base such as N-ethyl-morpholine or DiPEA and an activator such as TBTU or T3P, as shown in formula (VIII): [ka] [In the formula, PG represents a protecting group (e.g., tert-butoxycarbonyl), and Hal represents a halogen (e.g., I or Br)] The compound shown can be produced by reacting it with an aqueous ammonia solution. The reaction is preferably carried out in an organic solvent such as DMF at a temperature in the range of -20°C to 100°C, for example at 0°C.

[0110] The compound represented by formula (VIII) is either commercially available, known from literature (e.g., Tetrahedron: Asymmetry, 1998, 9, 503), or can be manufactured using known techniques.

[0111] Furthermore, a method for producing the compound represented by formula (I) defined above or a pharmaceutically acceptable salt thereof, using a standard procedure described in the literature for the dehydration of amides, for example, at a temperature in the range of -20°C to 100°C, for example at 25°C, in a solvent such as DCM or DMF, with or without a base such as DiPEA, for formula (IX): [ka] [In the formula, R 1is as defined above, and PG represents a protecting group (e.g., tert-butoxycarbonyl) There is provided a method comprising reacting a compound represented by with a reagent such as a Vilsmeier reagent or T3P, and then reacting with a reagent suitable for removing the protecting group PG. An example of a suitable reagent is formic acid.

[0112] The compound represented by formula (IX) is, in the presence of a catalyst such as bis[bis(1,2-diphenylphosphino)ethane]palladium(0) or Pd(dppf)Cl2·DCM and a base such as potassium carbonate or sodium carbonate, of formula (X):

Chemical formula

Chemical formula

[0113] The compound represented by formula (X) is, at a temperature within the range of 60 °C to 100 °C, for example 85 °C, in a solvent such as DMSO, with a suitable salt such as potassium acetate, with or without using 1,1'-bis(diphenylphosphino)ferrocene or palladium dichloride 1,1-bis(di-tert-butylphosphino)ferrocene, in the presence of a suitable catalyst such as Pd(dppf)Cl2·DCM, of formula (XII):

Chemical formula

[0114] The compound represented by formula (XII) is obtained by reacting the compound represented by formula (XIII): in the presence of a base such as DiPEA or TEA and an activator such as EDCI, 2-pyridinol-1-oxide or T3P.

Chemical formula

Chemical formula

[0115] The compound represented by formula (XIII) can be produced by using a standard procedure described in the literature for amide formation, for example, in the presence of a base such as N-ethyl-morpholine or DiPEA and an activator such as an "uronium" reagent (e.g., TBTU) or T3P, by reacting the compound represented by formula (XIV):

Chemical formula

[0116] The compound represented by formula (IX) can be produced by reacting the compound represented by formula (XII) [wherein PG represents a protecting group (e.g., tert-butoxycarbonyl)] with the compound represented by formula (VI) or its boronic acid ester in the presence of a catalyst such as bis[bis(1,2-diphenylphosphinol)]palladium(O) or Pd(dppf)Cl2·DCM and a base such as potassium carbonate or sodium carbonate. The reaction is conveniently carried out in a solvent such as dioxane / water or ACN / water mixture at a temperature in the range of 20°C to 100°C, particularly at 80°C.

[0117] Furthermore, a method for producing the compound represented by formula (I) defined above or a pharmaceutically acceptable salt thereof, wherein the compound is produced in the presence of a catalyst such as Pd(dppf)Cl2·DCM or 1,1-bis(di-tert-butylphosphin)ferrocenepalladium dichloride and a base such as potassium carbonate or sodium carbonate, wherein the compound is produced by formula (XV): [ka] [In the formula, PG represents a protecting group (e.g., tert-butoxycarbonyl)] The compound shown by formula (VI) [wherein R 1 A method is provided which involves reacting a compound or its ester as defined above. The reaction is conveniently carried out in a solvent such as a dioxane / water mixture or an ACN / water mixture at a temperature in the range of 20°C to 100°C, particularly at 75°C, and then reacted with a reagent suitable for removing the protecting group PG. An example of a suitable reagent is formic acid.

[0118] The compound represented by formula (XV) can be prepared from the compound represented by formula (XII) using standard literature-described procedures for amide dehydration, for example, at a temperature in the range of -20°C to 100°C, for example at 25°C, in a solvent such as DCM or DMF, with or without a base such as DiPEA, and using a reagent such as Burgess reagent, TBTU, or T3P.

[0119] Furthermore, a method for producing a compound represented by formula (I) as defined above or a pharmaceutically acceptable salt thereof, conveniently, in the presence of a base such as DiPEA or TEA and more than one activator such as EDCI, 2 - pyridinol -1-one or T3P, next, in the presence of a dehydrating reagent such as T3P, formula (XVI):

Chemical formula

[0120] The compound represented by formula (XVI) is obtained by reacting a compound represented by formula (VII) with a compound represented by formula (VI) [wherein, R 1 is as defined in formula (I)] or an ester thereof in the presence of a catalyst such as Pd(dppf)Cl2·DCM or 1,1 - bis(di - tert - butylphosphino) ferrocene palladium dichloride and a base such as potassium carbonate or sodium carbonate. The reaction is conveniently carried out in a solvent such as a dioxane / water mixture or an ACN / water mixture at a temperature within the range of, for example, 20 °C to 100 °C, particularly at 75 °C, and then the deprotection of PG is carried out.

[0121] Formula (III):

Chemical formula

Chemical formula

[0122] The compound represented by formula (XVII) [wherein PG represents a protecting group (e.g., tert-butoxycarbonyl)] is also represented by formula (XVIII): [ka] The compounds shown can be prepared by using a reducing agent, such as BH3-DMS, in a solvent such as THF at a temperature in the range of 0 to 40°C, for example at 25°C.

[0123] The compound represented by formula (XVIII) [wherein PG represents a protecting group (e.g., tert-butoxycarbonyl)] is also represented by formula (XIX): [ka] The compounds shown can be produced by using biocatalytic conversion for chemoselective lactam formation, for example, at a temperature in the range of 0 to 80°C, for example at 55°C, in a solvent such as ether (e.g., dioxane), using a lipase such as novozyme 435, and then by using conditions for the introduction of the protecting group PG.

[0124] The compound represented by formula (XIX) is represented by formula (XX): [ka] [In the formula, PG 1 and PG 2 [This represents a protecting group (e.g., benzyl)] The compounds shown can be prepared by using conditions for hydrogenation, for example, at a temperature in the range of 25-80°C, for example at 40°C, and under a pressure of, for example, 10 bar, in a solvent such as methanol or dioxane, using H2(g) and a reagent such as palladium-carbon dihydrate.

[0125] Formula (XX) [wherein, PG 1 and PG 2 Compounds represented by [where represents a protecting group (e.g., benzyl)] are given by formula (XXI): [ka] [In the formula, PG 1 and PG 2 [This represents a protecting group (e.g., benzyl)] The compound shown can be produced by reacting methyl propynate with methyl propynate in a solvent such as toluene and in the presence of a base such as 4-methylmorpholine at a temperature in the range of 0 to 100°C, for example at 25°C, using conditions for the oxa-michael reaction.

[0126] Formula (XXI) [wherein, PG 1 and PG 2 Compounds represented by [where represents a protecting group (e.g., benzyl)] can be produced by reacting a diprotected benzylamine (e.g., dibenzylamine) with (S)-oxirane-2-carboxylate methyl in a solvent such as ethanol at a temperature in the range of 0 to 78°C, for example at 70°C.

[0127] Alternatively, equation (III): [ka] [In the formula, PG represents a protecting group (e.g., tert-butoxycarbonyl)] The compound shown can be prepared, for example, at a temperature in the range of 0 to 100°C, for example at 25°C, in a solvent such as DCM / water, in which salts such as sodium bromide may be present, in the presence of a buffer such as NaHCO3 and a phase transfer catalyst such as tetrabutylammonium biphosphate, using reagents such as TEMPO and sodium hypochlorite, according to formula (XXII): [ka] It can be produced by oxidation of the compound shown.

[0128] The compound represented by formula (XXII) [wherein PG represents a protecting group (e.g., tert-butoxycarbonyl)] is also represented by formula (XXIII): [ka] [In the formula, PG 1 and PG 2 [This represents a protecting group (e.g., benzyl)] The compounds shown are reacted with a base such as sodium hydride in a solvent such as THF at a temperature in the range of 0 to 60°C, for example at 25°C, and then the protecting groups PG and PG defined in formulas (XXII) and (XXIII) are obtained. 1 and PG 2 It can be manufactured through mutual conversion.

[0129] Formula (XXIII) [wherein, PG 1 and PG 2 Compounds represented by [where represents a protecting group (e.g., benzyl)] can be produced by reacting a protected 3-aminopropanol (e.g., N-benzyl-3-aminopropanol) with (S)-2-((benzyloxy)methyl)oxirane in a solvent such as ethanol or propanol at a temperature in the range of 0 to 70°C, for example at 40°C, and then reacting the crude product with methanesulfonyl chloride in a solvent such as DCM in the presence of a base such as DiPEA at a temperature in the range of -10 to 25°C, for example at -5°C.

[0130] The compound represented by formula (VI) or its esters, as well as the compounds represented by formulas (VIII), (XI), and (XIV), are commercially available, known from the literature, or can be produced using known techniques.

[0131] Those skilled in the art will understand that in the manufacturing method of the present invention, certain functional groups in the reagent, such as hydroxyl or amino groups, may require protection by protecting groups. Thus, the production of the compound represented by formula (I) involves removing one or more protecting groups at an appropriate stage.

[0132] Those skilled in the art will understand that at any stage of the preparation of the compound represented by formula (I), a mixture of isomers (e.g., a racemate) of the compound corresponding to any of the compounds of formulas (II)-(V), (VII)-(X), and (XXII)-(XVI) may be available. At any stage of the preparation, one stereoisomer can be obtained by isolating from the mixture of isomers (e.g., a racemate) using, for example, chiral chromatography separation.

[0133] The protection and deprotection of functional groups are described in 'Protective Groups in Organic Synthesis', 4th Ed, TW Greene and PGM Wuts, Wiley (2006) and 'Protecting Groups', 3rd Ed, PJ Kocienski and Georg Thieme Verlag (2005).

[0134] Further embodiments include pharmaceutically acceptable salts of the compound represented by formula (I).

[0135] Salts of the compound represented by formula (I) may be advantageous due to one or more of their chemical or physical properties, such as stability at different temperatures and humidity levels, or desirable solubility in H2O, oil, or other solvents. In some cases, the salt can be used to aid in the isolation or purification of the compound. In some embodiments (particularly when the salt is intended for administration to animals (e.g., humans), or when the salt is a reagent used in the manufacture of a compound or salt intended for administration to animals), the salt is pharmaceutically acceptable.

[0136] When the compound represented by formula (I) is sufficiently acidic, pharmaceutically acceptable salts include, but are not limited to, alkali metal salts, such as Na or K, alkaline earth metal salts, such as Ca or Mg, or organic amine salts. When the compound represented by formula (I) is sufficiently basic, pharmaceutically acceptable salts include, but are not limited to, inorganic acid addition salts or organic acid addition salts.

[0137] Depending on the number of charged functional groups and the valence of the cation or anion, there may be two or more cations or anions.

[0138] For a discussion of suitable salts, see Berge et al., J. Pharm. Sci., 1977, 66, 1-19 or “Handbook of Pharmaceutical Salts: Properties, selection and use”, PH Stahl, PG Vermuth, IUPAC, Wiley-VCH, 2002.

[0139] In salts, proton transfer occurs between the compound represented by formula (I) and the counterion of the salt. However, in some cases, proton transfer may not be complete, and therefore the solid is not a true salt. In such cases, the compound represented by formula (I) and the "co-former" molecule in the solid interact primarily through nonionic forces such as hydrogen bonds. It is accepted that proton transfer is actually a continuum and can vary with temperature, and therefore the point at which it is more desirable for a salt to be described as a cocrystal is somewhat subjective.

[0140] If an acid or base coformer is solid at room temperature, and there is little to no proton transfer between the compound represented by formula (I) and such acid or base coformer, a cocrystal of the coformer and the compound represented by formula (I) may form rather than a salt. All such cocrystals of the compound represented by formula (I) are encompassed by the present invention.

[0141] The compound represented by formula (I) can form a mixture of its salt and cocrystal. The present invention should be understood to encompass the salt / cocrystal mixture of the compound represented by formula (I).

[0142] Salts and cocrystals can be characterized using well-known techniques, such as powder X-ray diffraction, single-crystal X-ray diffraction (e.g., to evaluate proton positions, bond lengths, or bond angles), solid-state NMR (e.g., to evaluate C, N, or P chemical shifts), or spectroscopic techniques (e.g., to measure OH, NH, or COOH signals and IR peak shifts resulting from hydrogen bonding).

[0143] Certain compounds represented by formula (I) may exist in solvated form, for example, as hydrates, and should be understood to include solvates of pharmaceutically acceptable salts of compounds represented by formula (I).

[0144] In further embodiments, compounds represented by certain formulas (I) may exist as racemates and racemic mixtures, single enantiomers, and individual diastereomers and diastereomer mixtures. The present invention should be understood to encompass all such isomers. Compounds represented by certain formulas (I) may also contain linking groups (e.g., carbon-carbon bonds, carbon-nitrogen bonds such as amide bonds), where bond rotation is limited with respect to the particular linking group, for example, by the presence of ring bonds or double bonds. Thus, the present invention should be understood to encompass all such isomers. Compounds represented by certain formulas (I) may also contain multiple tautomers. The present invention should be understood to encompass all such tautomers. Stereoiomers can be separated using conventional techniques, such as chromatography or fractional crystallization, or they can be produced by stereoselective synthesis.

[0145] In further embodiments, the compounds represented by formula (I) encompass isotopically labeled (or "radioactively labeled") derivatives of any of the compounds represented by formula (I). Such derivatives are derivatives of the compounds represented by formula (I) in which one or more atoms are replaced with atoms having atomic masses or mass numbers different from those typically found in nature. Examples of radionuclides that can be incorporated include: 2 One example is H (deuterium is sometimes written as "D").

[0146] In further embodiments, the compound represented by formula (I) may be administered in the form of a prodrug that is broken down in the body of a human or animal to become the compound represented by formula (I). An example of a prodrug is an in vivo hydrolyzable ester of the compound represented by formula (I).

[0147] In vivo hydrolyzable (cleavable) esters of compounds represented by formula (I) containing a carboxyl group or a hydroxyl group are pharmaceutically acceptable esters that hydrolyze in the body of humans or animals to produce hydrophilic or hydrophilic alcohols, for example. For examples of ester prodrug derivatives, see Curr. Drug. Metab. 2003, 4, 461.

[0148] Various other forms of prodrugs are known in the art. For examples of prodrug derivatives, see Nature Reviews Drug Discovery 2008, 7, 255 and the literature cited therein. [Examples]

[0149] The present invention will be further described here by reference to the following non-limiting embodiments.

[0150] (i) Unless otherwise specified, 1 ¹H NMR spectra were recorded using a Bruker Avance III spectrometer operated at electric field strengths of 400, 500, or 600 MHz. (Chloroform-d(CDCl3;δ)) H 7.27 ppm), Dimethyl sulfoxide-d6 (d3-DMSO; δ H 2.50 ppm) or methanol-d4 (CD3OD;δ H The central peak (3.31 ppm) was used as a reference.

[0151] (ii) MS spectra were recorded using either a Micromass ZQ single quadrupole LC-MS or Quattro Micro LC-MS following analytical HPLC using a Phenomenex Luna 5μ C18(2), 100×4.6 mm (plus guard cartridge) column and a gradient solution of 0.1% formic acid-containing ACN in a 0.1% formic acid aqueous solution, or a Waters Xterra MS 5μ C18, 100×4.6 mm (plus guard cartridge) column and a gradient solution of 10 mM ammonium bicarbonate aqueous solution of ACN. Ionization was, as specified, ESCI, with the option to obtain both ESI and APCI data in a single operation. Alternatively, LC-MS experiments were performed using a Waters Acquity UPLC system combined with a Waters Xevo Q-ToF Mass Spectrometer in ESI mode. The UPLC system is equipped with both a BEH C18 column (1.7 μm, 2.1 × 50 mm) paired with 46 mM ammonium carbonate / NH3 buffer (pH 10) and an HSS C18 column (1.8 μm, 2.1 × 50 mm) paired with 10 mM formic acid and 1 mM ammonium formate buffer (pH 3). Where m / z values ​​are available, generally only the ion indicating the mass is listed, and the cited mass ion is either positive or negative: [M] + [M+H] + [MH] - Or [M+2H-BOC] + .

[0152] (iii) The compounds in the titles and subtitles of the Examples and Preparation Examples were named using the IUPAC naming program ACD / Name2012 from Acdlabs.

[0153] (iv) Unless otherwise noted, the starting materials were commercially available, and all solvents and commercially available reagents were for experimental use and used as obtained. Unless otherwise noted, the operations were carried out at ambient temperature, i.e., in the range of 17–28°C, and, if necessary, under an atmosphere of an inert gas such as nitrogen.

[0154] (iv) X-ray diffraction analysis was performed according to standard methods, which can be found, for example, in Kitaigorodsky, AI (1973), Molecular Crystals and Molecules, Academic Press, New York; Bunn, CW (1948), Chemical Crystallography, Clarendon Press, London; or Klug, HP & Alexander, LE (1974), X-ray Diffraction Procedures, John Wiley & Sons, New York.

[0155] The sample was mounted on a single-crystal silicon (SSC) wafer mount, and powder X-ray diffraction was recorded using a PANalytical X'Pert PRO (reflection position, X-ray wavelength 1.5418 Å, nickel-filtered Cu irradiation, voltage 45 kV, filament emission 40 mA). The sample was rotated during the measurement using an auto-variable divergence and anti-scatter slits. The sample was scanned using a PIXCEL detector (effective length 3.35°2θ) with a step size of 0.013° and count times of 116 or 233 seconds from 2 to 50°2θ.

[0156] It is known in the art that powder X-ray diffraction patterns with one or more measurement errors can be obtained depending on the measurement conditions (e.g., the apparatus, sample preparation, or machine used). In particular, it is generally known that the intensity of the powder X-ray diffraction pattern can vary depending on the measurement conditions and sample preparation. For example, those skilled in the art of powder X-ray diffraction fully understand that the relative intensity of peaks can vary depending on the orientation of the sample under test and the type and setup of the instrument used. Those skilled in the art also fully understand that the position of reflections can be affected by the precise height at which the sample is placed in the diffractometer and the zero calibration of the diffractometer. The flatness of the sample surface can also have a small effect. Therefore, those skilled in the art will fully understand that the diffraction pattern data presented herein should not be interpreted as absolute, and that crystals that give substantially the same powder diffraction pattern as those described herein fall within the scope of the present invention (for further information, see Jenkins, R & Snyder, RL 'Introduction to X-Ray Powder Diffractometry' John Wiley & Sons, 1996).

[0157] In general, the measurement error of the diffraction angle in powder X-ray diffractograms can be approximately ±0.1°²θ, and the degree of such measurement error should be considered when examining powder X-ray diffraction data. Furthermore, it should be understood that the intensity varies depending on the experimental conditions and the treatment preparation (e.g., desired orientation). The following definitions were used for relative intensity (%): 81-100%, vs (very strong); 41-80%, str (strong); 21-40%, med (moderate); 10-20%, w (weak); 1-9%, vw (very weak).

[0158] The following abbreviations are used: [Table 1]

[0159] Production of boronic acid ester intermediates

[0160] Boronic acid ester 1 5-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-3,7-dimethyl-1,3-benzoxazole-2(3H)-one i) 5-Chloro-7-methyl-1,3-benzoxazole-2(3H)-one CDI (3.09 g, 19.0 mmol) was added to a solution of 2-amino-4-chloro-6-methylphenol (2.5 g, 15.9 mmol) in THF (65 mL). The reaction was heated under reflux for 2.5 hours and then cooled to room temperature. The reaction mixture was transferred to a separatory funnel and diluted with RINKAN (100 mL). The mixture was successively washed with 2 M hydrochloric acid, saturated sodium bicarbonate aqueous solution, and saturated sodium chloride solution. The organic extract was dried (sodium sulfate), filtered, and concentrated under reduced pressure to obtain the subtitle compound as a light brown solid (2.89 g, 98%). 1 H NMR (400 MHz, DMSO-d6): 11.84 (s, 1H), 7.10 (d, 1H), 7.06 (d, 1H), 2.37 (s, 3H).

[0161] ii) 5-Chloro-3,7-dimethyl-1,3-benzoxazole-2(3H)-one Cesium carbonate (2.65 g, 8.12 mmol) was added to a solution of 5-chloro-7-methyl-1,3-benzoxazole-2(3H)-one (1.50 g, 8.12 mmol) in DMF (100 mL). After 20 minutes, methyl iodide (0.61 mL, 9.84 mmol) was added dropwise, and the mixture was stirred at room temperature for 2 hours before being poured into ice water (100 mL). The resulting brown precipitate was collected by filtration and dried in a vacuum oven to obtain the compound of the subtitle as a brown solid (1.6 g, 100%). 1 1H NMR (400 MHz, DMSO-d6): 7.27 (s, 1H), 7.07 (s, 1H), 2.31 (s, 3H) (one CH3 under a weak peak).

[0162] iii) 5-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-3,7-dimethyl-1,3-benzoxazole-2(3H)-one To a solution of 5-chloro-3,7-dimethyl-1,3-benzoxazole-2(3H)-one (200 mg, 1.01 mmol) in 1,4-dioxane (5 mL), bis(neopentyl glycolate)diborone (342 mg, 1.52 mmol) and potassium acetate (198 mg, 2.02 mmol) were added. After degassing the reaction mixture under nitrogen for 15 minutes, XPhos (19 mg, 0.040 mmol) and chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (XPhos-Pd-G2, 16 mg, 0.020 mmol) were added. The reaction mixture was heated at 80°C for 3 hours. Subsequently, the reaction mixture was concentrated under reduced pressure, purified by silica gel column chromatography, and eluted with 0-20% siRNA in isohexane to obtain the labeled compound as a light brown oil (184 mg, 66%). 1 H NMR (400 MHz, CDCl3): δ 7.44 (s, 1H), 7.23 (s, 1H), 3.80 (s, 4H), 3.40 (s, 3H), 2.39 (s, 3H), 1.04 (s, 6H).

[0163] Boronic acid ester 2 7-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-1-methylquinoxaline-2(1H)-one To a solution of 7-bromo-1-methylquinoxaline-2(1H)-one (1.0 g, 4.2 mmol) in 1,4-dioxane (15 mL), bis(neopentyl glycolate)diborone (1.42 mg, 6.30 mmol) and potassium acetate (823 mg, 8.40 mmol) were added. The reaction mixture was degassed under nitrogen for 30 minutes, and then Pd(dppf)Cl2·DCM (171 mg, 0.21 mmol) was added. The reaction mixture was heated at 80°C for 3 hours. Subsequently, the reaction mixture was concentrated under reduced pressure, purified by silica gel column chromatography, and eluted with 30% siRNA in isohexane to obtain an orange solid. Trituration with diethyl ether yielded the marked compound as an off-white solid (340 mg, 30%). 1 H NMR (400 MHz, CDCl3): δ 8.37-8.30 (m, 1H), 7.79 (m, 3H), 3.82 (s, 4H), 3.75 (s, 3H), 1.06 (s, 6H).

[0164] Boronic acid ester 3 5-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-3-ethyl-1,3-benzoxazole-2(3H)-one i) 5-bromo-3-ethyl-1,3-benzoxazole-2(3H)-one Cesium carbonate (1.79 g, 5.5 mmol) was added to a solution of 5-bromo-1,3-benzoxazole-2(3H)-one (1.07 g, 5.0 mmol) in DMF (10 mL). Ethyl iodide (0.44 mL, 5.5 mmol) was added dropwise, and the reaction was stirred at room temperature for 24 hours. The solvent was removed under reduced pressure, and the resulting oil was dissolved in ELISA. The organic extract was successively washed with water and saturated sodium chloride solution, dried (magnesium sulfate), filtered, and concentrated under reduced pressure. The resulting oil was purified by silica gel column chromatography and eluted with DCM:isohexane in a ratio of 1:2 to obtain the subtitle compound as a white solid (1.06 g, 88%). 1H NMR (400 MHz, CDCl3): δ 7.25 (dd, 1H), 7.13 (d, 1H), 7.08 (d, 1H), 3.87 (dd, 2H), 1.39 (t, 3H).

[0165] ii) 5-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-3-ethyl-1,3-benzoxazole-2(3H)-one To a solution of 5-bromo-3-ethyl-1,3-benzoxazole-2(3H)-one (600 mg, 2.48 mmol) in 1,4-dioxane (10 mL), bis(neopentyl glycolate)diborone (616 mg, 2.73 mmol) and potassium acetate (487 mg, 4.96 mmol) were added. The reaction mixture was degassed under nitrogen for 30 minutes, and then Pd(dppf)Cl2·DCM (101 mg, 0.12 mmol) was added. The reaction mixture was heated at 80°C for 4 hours. Subsequently, the reaction mixture was concentrated under reduced pressure, purified by silica gel column chromatography, and eluted with 0-20% siRNA in isohexane to obtain the marked compound as an off-white solid (338 mg, 57%). 1 H NMR (400 MHz, CDCl3): δ 7.60 (d, 1H), 7.41 (s, 1H), 7.19 (d, 1H), 3.90 (dd, 2H), 3.79 (s, 4H), 1.43-1.35 (m, 3H), 1.04 (s, 6H).

[0166] Boronic acid ester 4 5-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-3-ethyl-7-methyl-1,3-benzoxazole-2(3H)-one i) 5-Chloro-3-ethyl-7-methyl-1,3-benzoxazole-2(3H)-one Using 5-chloro-7-methyl-1,3-benzoxazole-2(3H)-one (step i of boronic acid ester 1), the compound was prepared according to step i of boronic acid ester 3, and the subtitle compound was obtained as a brown solid (258 mg, 82%). 1H NMR (400 MHz, CDCl3): δ 6.93 (s, 1H), 6.82 (d, 1H), 3.85 (q, 2H), 2.35 (s, 3H), 1.37 (t, 3H).

[0167] ii) 5-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-3-ethyl-7-methyl-1,3-benzoxazole-2(3H)-one Using 5-chloro-3-ethyl-7-methyl-1,3-benzoxazole-2(3H)-one, the compound was prepared according to step iii) of Boronic Acid Ester 1, and the marked compound was obtained as an orange solid (285 mg, 81%). 1 H NMR (400 MHz, CDCl3): δ 7.42 (s, 1H), 7.24 (s, 1H), 3.93-3.83 (m, 2H), 3.78 (s, 4H), 2.37 (s, 3H), 1.41-1.32 (m, 3H), 1.04 (s, 6H).

[0168] Boronic acid ester 5 5-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-3-(2-hydroxy-2-methylpropyl)-1,3-benzoxazole-2(3H)-one i) 5-bromo-3-(2-oxopropyl)-1,3-benzoxazole-2(3H)-one Using 5-bromo-1,3-benzoxazole-2(3H)-one and chloroacetone, the compound of the subtitle was prepared according to step i) of boronic acid ester 3, and the subtitle compound was obtained as a yellow solid (1.31 g, 94%). 1 H NMR (400 MHz, CDCl3): δ 7.28-7.24 (m, 1H), 7.11 (d, 1H), 6.93 (d, 1H), 4.59 (s, 2H), 2.31 (s, 3H).

[0169] ii) 5-bromo-3-(2-hydroxy-2-methylpropyl)-1,3-benzoxazole-2(3H)-one While stirring at 0°C, methylmagnesium chloride (1.62 mL, 4.87 mmol, 3M solution in THF) was added to a solution of 5-bromo-3-(2-oxopropyl)-1,3-benzoxazole-2(3H)-one (1.31 g, 4.87 mmol) in THF (20 mL). After 1 hour, a further amount of methylmagnesium chloride (0.81 mL, 2.43 mmol) was added. The reaction was heated to room temperature and stirred for 1 hour, then quenched with ammonium chloride (saturated aqueous solution). The reaction mixture was diluted with RINKAN, and the layers were separated. The organic extract was successively washed with water and saturated sodium chloride solution, dried (magnesium sulfate), filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography and eluted with RINKAN and isohexane to obtain the subtitle compound as a brown solid (428 mg, 31%). 1 H NMR (400 MHz, CDCl3): δ 8.26 (s, 1H), 7.28-7.23 (m, 1H), 7.16-7.03 (m, 1H), 7.00-6.89 (m, 1H), 3.86 (s, 2H), 1.61 (s, 6H).

[0170] iii) 5-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-3-(2-hydroxy-2-methylpropyl)-1,3-benzoxazole-2(3H)-one Using 5-bromo-3-(2-hydroxy-2-methylpropyl)-1,3-benzoxazole-2(3H)-one, the compound was prepared according to step ii) of Boronic Acid Ester 3, and the marked compound was obtained as an orange solid (269 mg, 56%). 1 H NMR (400 MHz, CDCl3): δ 8.53 (s, 1H), 7.62 (dd, 1H), 7.42 (d, 1H), 7.06 (d, 1H), 3.96 (s, 2H), 3.76 (s, 4H), 1.62 (s, 6H), 1.11-0.96 (m, 6H).

[0171] Boronic acid ester 6 5-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-7-fluoro-3-methyl-1,3-benzoxazole-2(3H)-one i) 5-bromo-7-fluoro-1,3-benzoxazole-2(3H)-one CDI (2.38 g, 14.70 mmol) was added to a solution of 2-amino-4-bromo-6-fluorophenol (2.5 g, 12.25 mmol) in THF (65 mL). The reaction was heated under reflux for 2.5 hours and then cooled to room temperature. The reaction mixture was transferred to a separatory funnel and diluted with SiO2 (100 mL). The mixture was successively washed with 2 M hydrochloric acid, saturated sodium bicarbonate solution, and saturated sodium chloride solution. The organic extract was dried (sodium sulfate), filtered, and concentrated under reduced pressure. The resulting dark brown solid was triturated with diethyl ether and isohexane to obtain the subtitle compound as a light brown solid (2.01 g, 71%). 1 H NMR (400 MHz, DMSO-d6): δ 12.14 (s, 1H), 7.38 (dd, 1H), 7.16-7.15 (m, 1H).

[0172] ii) 5-bromo-7-fluoro-3-methyl-1,3-benzoxazole-2(3H)-one A solution of 5-bromo-7-fluoro-1,3-benzoxazole-2(3H)-one (2.01 g, 8.74 mmol) in DMF (30 mL) was added dropwise to a suspension of sodium hydride (419 mg, 10.49 mmol, 60% dispersion in mineral oil) in DMF (50 mL) while stirring at 0°C. The reaction was heated to room temperature for 30 minutes, then recooled to 0°C. Methyl iodide (653 μL) was added dropwise, and the reaction was heated to room temperature. After 18 hours, the reaction was carefully quenched with water and transferred to a separatory funnel. The mixture was extracted with diethyl ether (×3). The organic extract was successively washed with saturated sodium chloride solution, dried (magnesium sulfate), filtered, and concentrated under reduced pressure. The obtained substance was triturated with diethyl ether and isohexane to obtain the compound of the subtitle as a light brown solid (1.38 g, 64%). 1H NMR (400 MHz, DMSO-d6): δ 7.49 (d, 1H), 7.44 (dd, 1H), 3.35 (s, 3H).

[0173] iii) 5-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-7-fluoro-3-methyl-1,3-benzoxazole-2(3H)-one To a solution of 5-bromo-7-fluoro-3-methyl-1,3-benzoxazole-2(3H)-one (1.38 g, 5.60 mmol) in 1,4-dioxane (20 mL), bis(neopentyl glycolate)diborone (1.39 g, 6.17 mmol) and potassium acetate (1.10 g, 11.20 mmol) were added. The reaction mixture was degassed under nitrogen for 15 minutes, and then Pd(dppf)Cl2·DCM (229 mg, 0.28 mmol) was added. The reaction mixture was heated at 80°C for 3 hours. Subsequently, the reaction mixture was concentrated under reduced pressure, purified by silica gel column chromatography, and eluted with 20% siRNA in isohexane to obtain the marked compound as a light brown solid (1.16 g, 75%). 1 H NMR (400 MHz, CDCl3): δ 7.36 (d, 1H), 7.19 (m, 1H), 3.78 (s, 4H), 3.42 (s, 3H), 1.03 (s, 6H).

[0174] Boronic acid ester 7 5-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-3-(2,2-difluoroethyl)-1,3-benzoxazole-2(3H)-one i) 5-bromo-3-(2,2-difluoroethyl)-7-fluoro-1,3-benzoxazole-2(3H)-one Using 5-bromo-7-fluoro-1,3-benzoxazole-2(3H)-one (step i of Boronic Acid Ester 6) and 2,2-difluoroethyltrifluoromethanesulfonate, the compound of the subtitle was prepared according to step i of Boronic Acid Ester 3, yielding the subtitle compound as a brown solid (2.49 g, 89%). 1H NMR (400 MHz, CDCl3): δ 7.16 (dd, 1H), 7.05 (s, 1H), 6.08 (tt, 1H), 4.16 (td, 2H).

[0175] ii) 5-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-3-(2,2-difluoroethyl-2-methylpropyl)-1,3-benzoxazole-2(3H)-one Using 5-bromo-3-(2,2-difluoroethyl)-7-fluoro-1,3-benzoxazole-2(3H)-one, the compound was prepared according to step ii) of Boronic Acid Ester 3, and the marked compound was obtained as an off-white solid (1.15 g, 41%). 1 H NMR (400 MHz, CDCl3): δ 7.40 (d, 1H), 7.30-7.24 (m, 1H), 6.10 (tt, 1H), 4.23-4.12 (m, 2H), 3.78 (s, 4H), 1.03 (s, 6H).

[0176] Boronic acid ester 8 5-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-3-(2-(dimethylamino)ethyl)-1,3-benzoxazole-2(3H)-one i) 5-bromo-3-(2-(dimethylamino)ethyl)-1,3-benzoxazole-2(3H)-one 5-bromo-1,3-benzoxazole-2(3H)-one (1.80 g, 8.41 mmol) and potassium carbonate (3.87 g, 28.0 mmol) in DMF (10 mL) were mixed with 2-dimethylaminoethyl chloride hydrochloride (1.21 g, 8.41 mmol). The reaction was heated at 125 °C for 3.5 hours, then cooled to room temperature and poured into ice water. The aqueous layer was extracted with SiO2 (100 mL x 4). The combined organic extract was dried (magnesium sulfate), filtered, and concentrated under reduced pressure. The resulting oil was dissolved in diethyl ether, washed with water, dried (magnesium sulfate), filtered, and concentrated under reduced pressure to obtain the compound of the subtitle as a light brown oil (1.63 g, 68%). 1H NMR (400 MHz, CDCl3): δ 7.23 (dd, 1H), 7.15 (d, 1H), 7.10-7.02 (m, 1H), 3.93-3.85 (m, 2H), 2.69-2.61 (m, 2H), 2.30 (s, 6H).

[0177] ii) 5-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-3-(2-(dimethylamino)ethyl)-1,3-benzoxazole-2(3H)-one Using 5-bromo-3-(2-(dimethylamino)ethyl)-1,3-benzoxazole-2(3H)-one, the compound was prepared according to step ii) of Boronic Acid Ester 3, and the marked compound was obtained as an off-white solid (1.15 g, 41%). It was used in the next step without further purification.

[0178] Boronic acid ester 9 6-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-3,3-difluoro-1-methyl-1,3-dihydro-2H-indole-2-one i) 6-bromo-3,3-difluoro-1,3-dihydro-2H-indole-2-one While stirring at room temperature, bis(2-methoxyethyl)aminosulfate trifluoride (deoxo-fluor, 44.25 mL, 22.12 mmol, 50% solution in THF) was added dropwise to a suspension of 6-bromoisatin (2.0 g, 8.75 mmol) in DCM (90 mL) over 30 minutes. After 24 hours, the reaction was carefully quenched at 0°C with a saturated sodium bicarbonate solution (40 mL). The aqueous layer was separated, the organic extract was dried (hydrophobic frit / phase separator), and concentrated under reduced pressure. The crude substance was purified by silica gel column chromatography and eluted with 20% siRNA in isohexane to obtain the subtitle compound as an orange solid (1.63 g, 74%). 1 ¹H NMR (400 MHz, CH3OH-d4): δ 7.50-7.46 (m, 1H), 7.36 (dd, 1H), 7.18 (d, 1H), (no exchangeable molecules were observed).

[0179] ii) 6-bromo-3,3-difluoro-1-methyl-1,3-dihydro-2H-indole-2-one Using 6-bromo-3,3-difluoro-1,3-dihydro-2H-indole-2-one and methyl iodide, the compound of the subtitle was prepared according to step i) of Boronic Acid Ester 3, and the subtitle compound was obtained as an orange solid (1.39 g, 82%). It was used in the next step without further purification.

[0180] iii) 6-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-3,3-difluoro-1-methyl-1,3-dihydro-2H-indole-2-one The compound was prepared using 6-bromo-3,3-difluoro-1-methyl-1,3-dihydro-2H-indole-2-one according to step ii) of Boronic Acid Ester 3, and the marked compound was obtained as an off-white solid (120 mg, 8%). 1 H NMR (400 MHz, CDCl3): δ 7.62 (t, 1H), 7.55-7.47 (m, 1H), 7.31-7.28 (m, 1H), 3.80 (s, 4H), 3.22 (s, 3H), 1.04 (s, 6H).

[0181] Boronic acid ester 10 3-(cyclopropylmethyl)-5-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-1,3-benzoxazole-2(3H)-one i) 5-bromo-3-(cyclopropylmethyl)-1,3-benzoxazole-2(3H)-one Cesium carbonate (1.79 g, 5.5 mmol) was added to a solution of 5-bromo-1,3-benzoxazole-2(3H)-one (1.07 g, 5.0 mmol) in DMF (10 mL). (Bromomethyl)cyclopropane (743 mg, 5.5 mmol) was added dropwise, and the reaction was stirred at room temperature for 24 hours. The solvent was removed under reduced pressure, and the resulting oil was dissolved in RINKAN. The organic extract was washed with water and a saturated sodium chloride solution, dried, filtered, and concentrated under reduced pressure. The resulting oil was purified by silica gel column chromatography and eluted with DCM:isohexane in a ratio of 1:2 to obtain the compound of the subtitle as a white solid (918 mg, 68%). 1 H NMR (400 MHz, CDCl3): δ 7.24 (dd, 1H), 7.17 (d, 1H), 7.13-7.04 (m, 1H), 3.68 (d, 2H), 1.29-1.17 (m, 1H), 0.69-0.54 (m, 2H), 0.51-0.41 (m, 2H).

[0182] ii) 3-(cyclopropylmethyl)-5-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-1,3-benzoxazole-2(3H)-one Using 5-bromo-3-(cyclopropylmethyl)-1,3-benzoxazole-2(3H)-one, the compound was prepared according to step ii) of Boronic Acid Ester 3, and the marked compound was obtained as a brown solid (520 mg, 62%). It was used in the next step without further purification.

[0183] Boronic acid ester 11 5-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-3-(2-methoxyethyl)-1,3-benzothiazole-2(3H)-one i) 5-Chloro-3-(2-methoxyethyl)-1,3-benzothiazole-2(3H)-one Using 5-chloro-1,3-benzothiazole-2(3H)-one and 1-bromo-2-methoxyethane, the compound of the subtitle was prepared according to step ii) of Boronic Acid Ester 1, and the subtitle compound was obtained as a yellow solid (3.5 g, 89%). 1 H NMR (400 MHz, CDCl3): δ 7.32 (d, 1H), 7.22 (d, 1H), 7.13 (dd, 1H), 4.12-4.06 (m, 2H), 3.71-3.65 (m, 2H), 3.34 (s, 3H).

[0184] ii) 5-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-3-(2-methoxyethyl)-1,3-benzothiazole-2(3H)-one Using 5-chloro-3-(2-methoxyethyl)-1,3-benzothiazole-2(3H)-one, the compound was prepared according to step iii) of Boronic Acid Ester 1, and the marked compound was obtained as a pale brown solid (1.02 g, 64%). It was used in the next step without further purification.

[0185] Boronic acid ester 12 5-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-3-isopropyl-1,3-benzoxazole-2(3H)-one i) 5-bromo-3-isopropyl-1,3-benzoxazole-2(3H)-one Using 5-bromo-1,3-benzoxazole-2(3H)-one and 2-iodopropane, the compound was prepared according to step i) of Boronic Acid Ester 3, and the subtitle compound was obtained as a white solid (510 mg, 66%). It was used in the next step without further purification.

[0186] ii) 5-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-3-isopropyl-1,3-benzoxazole-2(3H)-one Using 5-bromo-3-isopropyl-1,3-benzoxazole-2(3H)-one, the compound of the subtitle was prepared according to step ii) of boronic acid ester 3, as a brown solid (132 mg, 19%). It was used in the next step without further purification.

[0187] Boronic acid ester 13 6-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-4-methyl-2H-1,4-benzoxazine-3(4H)-one Using commercially available 6-bromo-4-methyl-2H-1,4-benzoxazine-3(4H)-one, the compound was prepared according to step ii) of Boronic Acid Ester 3, and the marked compound was obtained as a brown solid (520 mg, 62%). It was used directly without further purification.

[0188] Boronic acid ester 14 7-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-1-methylquinoline-2(1H)-one i) 7-Bromoquinoline-2(1H)-one A mixture of 7-bromo-2-chloroquinoline (5.0 g, 20.6 mmol) in a 5 M aqueous hydrochloric acid solution (133 mL) and 1,4-dioxane (14 mL) was stirred and heated under reflux for 2 hours. The reaction was cooled, the resulting precipitate was collected by filtration, washed with water, and the compound of the subtitle was obtained as a colorless solid (4.3 g, 93%). 1 H NMR (400 MHz, DMSO-d6): δ 11.80 (s, 1H), 7.91 (d, 1H), 7.63 (d, 1H), 7.48 (d, 1H), 7.34 (dd, 1H), 6.53 (d, 1H).

[0189] ii) 7-Bromo-1-methylquinoline-2(1H)-one Sodium hydride (320 mg, 7.98 mmol, 60% dispersion in mineral oil) was added to a solution of 7-bromoquinoline-2(1H)-one (1.5 g, 6.64 mmol) in anhydrous THF under a nitrogen atmosphere at room temperature with stirring. After 1 hour, the reaction mixture was cooled to 0°C, methyl iodide (1.88 g, 0.81 ml, 13.28 mmol) was added, and the reaction was slowly warmed to room temperature. After 18 hours, the reaction was carefully quenched with water (1 mL) and concentrated under reduced pressure. The resulting residue was partitioned into ethyl acetate and water. Each layer was separated, and the aqueous layer was extracted with ethyl acetate. The combined organic extract was dried (magnesium sulfate), filtered, and concentrated under reduced pressure. Isohexane was added to the residue, and recrystallization from DCM yielded the compound of the subtitle as a colorless solid (650 mg, 40%). 1 H NMR (400 MHz, CDCl3): δ 7.62 (d, 1H), 7.53 (d, 1H), 7.40 (s, 1H), 7.35 (dd, 1H), 6.75-6.66 (m, 1H), 3.69 (s, 3H).

[0190] ii) 7-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-1-methylquinoline-2(1H)-one Starting from 7-bromo-1-methylquinoline-2(1H)-one, the compound was prepared according to the procedure for boronic acid ester 2, and the marked compound was obtained as a pale pink solid (650 mg, 88%). 1 H NMR (400 MHz, CDCl3): δ 7.83 (s, 1H), 7.69-7.60 (m, 2H), 7.53 (d, 1H), 6.73 (d, 1H), 3.82 (s, 4H), 3.78 (s, 3H), 1.05 (s, 6H).

[0191] Boronic acid ester 15 5-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-3-(tetrahydro-2H-pyran-4-ylmethyl)-1,3-benzoxazole-2(3H)-one i) 5-bromo-3-(tetrahydro-2H-pyran-4-ylmethyl)-1,3-benzoxazole-2(3H)-one 5-bromo-2-benzoxazolinone (795 mg, 3.7 mmol) and cesium carbonate (500 mg, 7.4 mmol) in DMF (10 mL) were mixed with 4-(chloromethyl)tetrahydro-2H-pyran (500 mg, 3.7 mmol). The reaction was heated at 110 °C for 48 hours, then cooled to room temperature and poured into ice water. The resulting precipitate was collected by filtration and vacuum-dried to obtain the compound in question as a light brown oil (840 mg, 73%). 1 H NMR (400 MHz, DMSO-d6): δ 7.70 (s, 1H), 7.34-7.27 (m, 2H), 3.88-3.78 (m, 2H), 3.71 (d, 2H), 3.25 (td, 2H), 2.11-1.99 (m, 1H), 1.53 (d, 2H), 1.35-1.22 (m, 2H).

[0192] ii) 5-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-3-(tetrahydro-2H-pyran-4-ylmethyl)-1,3-benzoxazole-2(3H)-one Starting from 5-bromo-3-(tetrahydro-2H-pyran-4-ylmethyl)-1,3-benzoxazole-2(3H)-one, the compound was prepared according to the procedure for boronic acid ester 2, and the marked compound was obtained as an orange solid (440 mg, 47%). 1 ¹H NMR (400 MHz, CDCl3): δ 7.61 (dd, 1H), 7.37 (s, 1H), 3.98 (dd, 2H), 3.79 (s, 3H), 3.75-3.69 (m, 2H), 3.40-3.32 (m, 2H), 2.25-2.11 (m, 1H), 1.66-1.53 ​​(m, 3H), 1.53-1.39 (m, 2H), 1.04 (s, 6H) (one H under the CHCl3 peak).

[0193] Boronic acid ester 16 7-Chloro-5-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-3-methyl-1,3-benzoxazole-2(3H)-one i) 4-Bromo-2-chloro-6-nitrophenol At room temperature, 11.5 mL of 70% nitric acid aqueous solution (190 mol) was slowly added to a solution of 4-bromo-2-chlorophenol (20.0 g, 96.4 mmol) in acetic acid (100 mL). The resulting precipitate was collected by filtration, and the compound in question was obtained as a yellow solid (24.0 g). It was used in the next step without further purification.

[0194] ii) 2-amino-4-bromo-6-chlorophenol Calcium chloride (443 mg, 4 mmol) and iron (11.16 g, 0.2 mol) were added to a solution of 4-bromo-2-chloro-6-nitrophenol (10.0 g) in ethanol (400 mL) and water (100 mL). The suspension was heated at 80 °C for 2 hours. The reaction was cooled, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was diluted with saturated sodium chloride solution (500 mL) and extracted with ethyl acetate (500 mL x 2). The combined organic extract was dried (magnesium sulfate), filtered, and concentrated under reduced pressure to obtain the compound of the subtitle as a black solid (4 g, 45%). 1 H NMR (400 MHz, CDCl3): δ 6.85 (d, 1H), 6.75 (d, 1H), 5.38 (s, 1H), 3.92 (bs, 2H).

[0195] iii) 5-bromo-7-chloro-1,3-benzoxazole-2(3H)-one CDI (4.0 g, 24.6 mmol) was added to a stirred solution of 2-amino-4-bromo-6-chlorophenol (2.0 g, 9.0 mmol) in anhydrous THF (50 mL). The mixture was heated under reflux under a nitrogen atmosphere for 2.5 hours. The reaction was cooled, and the solvent was removed under reduced pressure. The resulting residue was washed with 2N hydrochloric acid aqueous solution, and then triturated with methanol to obtain the compound of the subtitle as a brown solid (0.8 g, 36%). 1H NMR (400 MHz, DMSO-d6): δ 12.18 (s, 1H), 7.46 (d, 1H), 7.27 (d, 1H).

[0196] iv) 5-bromo-7-chloro-3-methyl-1,3-benzoxazole-2(3H)-one Starting from 5-bromo-7-chloro-1,3-benzoxazole-2(3H)-one, and using potassium carbonate instead of cesium carbonate, the compound was prepared according to step ii) of Boronic Acid Ester 1, and the subtitle compound was obtained as a brown solid (700 mg, 83%). 1 H NMR (400 MHz, CDCl3): δ 7.26 (s, 1H), 7.02 (d, 1H), 3.40 (s, 3H).

[0197] v)7-Chloro-5-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-3-methyl-1,3-benzoxazole-2(3H)-one Starting from 5-bromo-7-chloro-3-methyl-1,3-benzoxazole-2(3H)-one, the compound was prepared according to the procedure for boronic acid ester 2, and the marked compound was obtained as an off-white solid (170 mg, 22%). 1 H NMR (400 MHz, CDCl3): δ 7.59 (s, 1H), 7.28 (s, 1H), 3.78 (s, 4H), 3.41 (s, 3H), 1.03 (s, 6H).

[0198] Boronic acid ester 17 3-(2,2-difluoroethyl)-5-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-1,3-benzoxazole-2(3H)-one i) 5-Chloro-3-(2,2-difluoroethyl)-1,3-benzoxazole-2(3H)-one To a solution of 5-chloro-1,3-benzoxazole-2(3H)-one (1 g, 5.89 mmol) in DMF (20 mL), cesium carbonate (3.83 g, 11.8 mmol) was added, followed by the dropwise addition of 2,2-difluoroethyltrifluoromethanesulfonate (1.38 g, 6.5 mmol). The resulting mixture was stirred at room temperature for 30 minutes. Then, water (60 mL) was added, and the resulting precipitate was collected by filtration, washed with water, and vacuum-dried to obtain the compound of the subtitle as a white solid (1.25 g, 91%). 1 H NMR (400 MHz, CDCl3): δ 7.24-7.18 (m, 1H), 7.08 (m, 1H), 7.01 (s, 1H), 6.17-5.85 (m, 1H), 4.14-4.04 (m, 2H).

[0199] ii) 3-(2,2-difluoroethyl)-5-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-1,3-benzoxazole-2(3H)-one Using 5-chloro-3-(2,2-difluoroethyl)-1,3-benzoxazole-2(3H)-one, the compound was prepared according to step iii) of Boronic Acid Ester 1, and the marked compound was obtained as an off-white solid (670 mg, 40%). 1 H NMR (400 MHz, CDCl3): δ 7.67-7.61 (m, 1H), 7.48 (s, 1H), 7.21 (d, 1H), 6.26-5.93 (m, 1H), 4.22-4.10 (m, 2H), 3.78 (s, 4H), 1.03 (s, 6H).

[0200] Boronic acid ester 18 3-(2,2,2-trifluoroethyl)-5-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-1,3-benzoxazole-2(3H)-one i) 5-Chloro-3-(2,2,2-trifluoroethyl)-1,3-benzoxazole-2(3H)-one To a solution of 5-chloro-1,3-benzoxazole-2(3H)-one (1 g, 5.89 mmol) in DMF (20 mL), cesium carbonate (3.83 g, 11.8 mmol) was added, followed by the addition of 2,2,2-trifluoroethyltrifluoromethanesulfonate (1.5 g, 6.5 mmol). The resulting mixture was stirred at room temperature for 30 minutes. Water (60 mL) was added, and the resulting precipitate was collected by filtration, washed with water, and vacuum-dried to obtain the compound of the subtitle as a white solid (1.31 g, 89%). 1 H NMR (400 MHz, CDCl3): δ 7.19-7.15 (m, 2H), 7.08 (s, 1H), 4.40 (q, 2H).

[0201] ii) 3-(2,2,2-trifluoroethyl)-5-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-1,3-benzoxazole-2(3H)-one Using 5-chloro-3-(2,2,2-trifluoroethyl)-1,3-benzoxazole-2(3H)-one, the compound was prepared according to step iii) of Boronic Acid Ester 1, and the marked compound was obtained as an off-white solid (670 mg, 40%). 1 H NMR (400 MHz, CDCl3): δ 7.67 (dd, 1H), 7.47 (s, 1H), 7.22 (d, 1H), 4.41 (dd, 2H), 3.78 (s, 4H), 1.03 (s, 6H).

[0202] Boronic acid ester 19 5-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-3-methyl-1,3-benzothiazole-2(3H)-one i) 5-Chloro-3-methyl-1,3-benzothiazole-2(3H)-one Cesium carbonate (17.5 g, 53.8 mmol) was added to a solution of 5-chloro-1,3-benzothiazole-2(3H)-one (5.0 g, 26.9 mmol) in DMF (70 mL). After 20 minutes, methyl iodide (2.51 mL, 40.4 mmol) was added dropwise. After the addition was complete, the reaction mixture was stirred at room temperature for 2 hours and then poured into ice water (300 mL). The resulting brown precipitate was collected by filtration and dried in a vacuum oven to obtain the compound of the subtitle as a colorless solid (4.42 g, 82%). 1 H NMR (400 MHz, CDCl3): δ 7.35 (d, 1H), 7.17 (dd, 1H), 7.06 (d, 1H), 3.45 (s, 3H).

[0203] ii) 5-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-3-methyl-1,3-benzothiazole-2(3H)-one Using 5-chloro-3-methyl-1,3-benzothiazole-2(3H)-one, the compound was prepared according to step iii) of Boronic Acid Ester 1, and the marked compound was obtained as an off-white solid (620 mg, 15%). 1 H NMR (400 MHz, CDCl3): δ 7.61 (dd, 1H), 7.50-7.36 (m, 2H), 3.80 (s, 4H), 3.48 (s, 3H), 1.04 (s, 6H).

[0204] Boronic acid ester 20 6-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-4-methyl-2H-1,4-benzothiazine-3(4H)-one i) 2-((4-bromo-2-nitrophenyl)thio)acetic acid Potassium carbonate (4.55 g, 33 mmol) and thioacetic acid (1.15 mL, 16.5 mmol) were successively added to a solution of 4-bromo-1-fluoro-2-nitrobenzene (3.02 g, 15 mmol) in DMF (20 mL) while stirring at room temperature. After 18 hours, the reaction was diluted with ethyl acetate and water. Each layer was separated. The aqueous layer was acidified and extracted with ethyl acetate. The organic extract was dried (magnesium sulfate), filtered, and concentrated under reduced pressure to obtain the compound of the subtitle as a yellow solid (2.60 g, 59%). 1 H NMR (400 MHz, DMSO-d6): δ 13.04 (s, 1H), 8.37 (d, 1H), 7.96-7.91 (m, 1H), 7.54 (d, 1H), 4.05 (s, 2H).

[0205] ii) 6-bromo-2H-1,4-benzothiazine-3(4H)-one At room temperature, iron(II) sulfate heptahydrate (18.12 g, 65.17 mmol) in water (25 mL) was slowly added to a solution of ammonium hydroxide (26 mL) and 2-((4-bromo-2-nitrophenyl)thio)acetic acid (2.6 g, 8.93 mmol). After 3 hours, the reaction mixture was filtered through Celite and washed with ammonium hydroxide and water. The filtrate was acidified with concentrated hydrochloric acid, and the resulting precipitate was collected by filtration. The solid was dissolved in ethyl acetate, dried (in magnesium sulfate), filtered, and concentrated under reduced pressure to obtain the compound of the subtitle as a yellow solid (1.9 g, 93%). 1 H NMR (400 MHz, DMSO-d6): δ 10.64 (br, 1H), 7.36-7.26 (m, 1H), 7.15 (dd, 2H), 3.49 (s, 2H).

[0206] iii) 6-bromo-4-methyl-2H-1,4-benzothiazine-3(4H)-one Starting with 6-bromo-2H-1,4-benzothiazine-3(4H)-one, the compound was prepared according to step ii) of Boronic Acid Ester 1, and the subtitle compound was obtained as a yellow solid (1.16 g, 86%). 1H NMR (400 MHz, CDCl3): δ 7.25 (d, 1H), 7.21 (d, 1H), 7.15 (dd, 1H), 3.42 (s, 3H), 3.40 (s, 2H).

[0207] iv) 6-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-4-methyl-2H-1,4-benzothiazine-3(4H)-one Using 6-bromo-4-methyl-2H-1,4-benzothiazine-3(4H)-one, the preparation was carried out according to step ii) of boronic acid ester 3, and the marked compound was obtained as a white solid (655 mg, 45%). 1 H NMR (400 MHz, CDCl3): δ 7.58-7.44 (m, 1H), 7.45 (dd, 1H), 7.38-7.30 (m, 1H), 3.77 (s, 4H), 3.48 (s, 3H), 3.47-3.35 (m, 2H), 1.03 (s, 6H).

[0208] Boronic acid ester 21 5-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-3-(2-methoxyethyl)-1,3-benzoxazole-2(3H)-one i) 5-bromo-3-(2-methoxyethyl)-1,3-benzoxazole-2(3H)-one Using 5-bromo-1,3-benzoxazole-2(3H)-one, the compound of the subtitle was prepared according to step i) of boronic acid ester 3, and the subtitle compound was obtained as a yellow solid (1.15 g, 85%). 1 H NMR (400 MHz, CDCl3): δ 7.27-7.20 (m, 2H), 7.06 (d, 1H), 3.97 (t, 2H), 3.72-3.66 (m, 2H), 3.35 (s, 3H).

[0209] ii) 5-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-3-(2-methoxyethyl)-1,3-benzoxazole-2(3H)-one Using 5-bromo-3-(2-methoxyethyl)-1,3-benzoxazole-2(3H)-one, the compound was prepared according to step ii) of Boronic Acid Ester 3, and the marked compound was obtained as a yellow oil (1.15 g, 85%). It was used in the next step without further purification.

[0210] Boronic acid ester 22 5-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-3-methyl-1,3-benzoxazole-2(3H)-one [ka]

[0211] i) 5-Chloro-3-methyl-1,3-benzoxazole-2(3H)-one [ka] Cesium carbonate (19.21 g, 58.96 mmol) was added to a solution of 5-chloro-1,3-benzoxazole-2(3H)-one (10 g, 58.96 mmol) in DMF (100 mL). After 30 minutes, methyl iodide (4.40 mL, 70.75 mmol) was added dropwise. After the addition was complete, the reaction mixture was stirred at room temperature for 18 hours and then poured into ice water (500 mL). The resulting white precipitate was collected by filtration and dried in a vacuum oven over P2O5 to obtain the compound of the subtitle as a white solid (9.92 g, 92%). 1 H NMR (400 MHz, DMSO-d6): δ 7.46 (d, 1H), 7.36 (d, 1H), 7.17 (dd, 1H), 3.34 (s, 3H).

[0212] ii) 5-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-3-methyl-1,3-benzoxazole-2(3H)-one To a solution of 5-chloro-3-methyl-1,3-benzoxazole-2(3H)-one (3.0 g, 16.3 mmol) in 1,4-dioxane (80 mL), bis(neopentyl glycolate)diborone (5.54 g, 24.5 mmol) and potassium acetate (3.21 g, 32.7 mmol) were added. After degassing the reaction mixture under nitrogen for 40 minutes, XPhos (311 mg, 0.65 mmol) and chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (XPhos-Pd-G2, 257 mg, 0.33 mmol) were added. The reaction mixture was heated at 80°C for 2 hours. Subsequently, the reaction mixture was concentrated under reduced pressure, purified by silica gel column chromatography, and eluted with 0-10% dimethyl ammonium compounds in isohexane to obtain the labeled compound as a yellow solid (4.8 mg, >100%). 1 H NMR (400 MHz, CDCl3): δ 7.61 (dd, 1H), 7.40 (s, 1H), 7.21-7.13 (m, 1H), 3.79 (s, 4H), 3.41 (s, 3H), 1.04 (s, 6H).

[0213] Manufacturing of intermediate components

[0214] Intermediate 1 4'-[(2S)-2-amino-2-cyanoethyl]biphenyl-4-carbonitrile i) [(1S)-1-cyano-2-(4'-cyanobiphenyl-4-yl)ethyl] tert-butyl carbamate Potassium carbonate (4.5 g, 36 mmol) was added to a suspension of tert-butyl N-[(1S)-1-cyano-2-(4-iodophenyl)ethyl]carbamate (prepared according to the procedure on page 47 of International Publication No. 2009 / 74829) (5.99 g, 16 mmol) and (4-cyanophenyl)boronic acid (2.64 g, 18 mmol) in 1,4-dioxane (60 mL) and water (8 mL). The suspension was stirred under a nitrogen stream for 15 minutes, and then Pd(dppf)Cl2·DCM (1.3 g) was added. The reaction was heated at 75°C for 45 minutes and then concentrated under reduced pressure. The resulting oil was diluted with ELISA (200 mL) and washed with water (100 mL) and saturated sodium chloride solution (50 mL). The organic extract was dried over magnesium sulfate, filtered, and evaporated under reduced pressure to obtain a brown oil. The oil was purified by silica gel column chromatography and eluted with 20-30% siRNA in isohexane to obtain the compound in question as a colorless solid (5.9 g, 90%). 1 H NMR (400 MHz, DMSO-d6): δ 10.63 (s, 1H), 7.96-7.81 (m, 4H), 7.73 (d, 2H), 7.45 (d, 2H), 4.71 (q, 1H), 3.18-3.05 (m, 2H), 1.36 (s, 9H).

[0215] ii) 4'-[(2S)-2-amino-2-cyanoethyl]biphenyl-4-carbonitrile [(1S)-1-cyano-2-(4'-cyanobiphenyl-4-yl)ethyl]carbamate tert-butyl (5.4 g, 15.5 mmol) was dissolved in formic acid (50 mL) and heated to 50°C for 15 minutes on a preheated hot plate stirrer. The solution was evaporated under reduced pressure and diluted with toluene (150 mL). A saturated aqueous solution of sodium bicarbonate was added to make the mixture basic (pH 8). The toluene was separated, washed with saturated sodium chloride, dried (magnesium sulfate), filtered, and evaporated under reduced pressure to obtain a yellow oil. The oil was purified by silica gel column chromatography and eluted with toluene to obtain the marked compound as a colorless solid (2.88 g, 74%). 1¹H NMR (400 MHz, CDCl3): δ 7.62 (m, 4H), 7.52 (m, 2H), 7.35 (d, 2H), 3.92 (t, 1H), 3.10-2.96 (m, 2H) (two exchangeable protons were not observed).

[0216] Intermediate 2 (2S)-2-amino-3-[4-(3-methyl-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl)phenyl]propannitrile i) {(1S)-1-cyano-2-[4-(3-methyl-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl)phenyl]ethyl} tert-butyl carbamate} [ka] 5-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-3-methyl-1,3-benzoxazole-2(3H)-one (boronic acid ester 22, 3.34 g, 12.81 mmol) and (S)-(1-cyano-2-(4-iodophenyl)ethyl)carbamate tert-butyl (prepared according to the procedure on page 47 of International Publication No. 2009 / 074829) (12.81 mmol) were dissolved in 1,4-dioxane (340 mL) and water (12 mL). The reaction mixture was degassed under nitrogen for 30 minutes, after which potassium carbonate (2.66 g, 19.21 mmol) and Pd(dppf)Cl2·DCM (1.05 g, 1.28 mmol) were added. The reaction mixture was heated at 80°C for 1.5 hours. The reaction was then concentrated under reduced pressure. The residue was diluted with toluene (200 mL) and water (50 mL). The mixture was filtered through Celite, and each layer was separated. The organic extract was washed with a saturated sodium chloride solution, dried (magnesium sulfate), filtered, and evaporated. The resulting oil was purified by silica gel column chromatography and eluted with a 0-40% toluene gradient solution in isohexane to obtain the subtitle compound as a white solid (3.87 mg, 77%). 1H NMR (400 MHz, CDCl3): δ 7.51-7.46 (m, 2H), 7.31 (d, 2H), 7.21-7.17 (m, 3H), 7.12-7.06 (m, 1H), 4.78 (s, 1H), 3.39 (s, 3H), 3.14-2.98 (m, 2H), 1.39 (s, 9H).

[0217] ii) (2S)-2-amino-3-[4-(3-methyl-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl)phenyl]propannitrile [ka] {(1S)-1-cyano-2-[4-(3-methyl-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl)phenyl]ethyl}carbamate tert-butyl (3.87 g, 9.84 mmol) was mixed with formic acid (32 mL). The mixture was heated at 50°C for 15 minutes on a preheated hot plate stirrer. The solvent was then removed under reduced pressure. The residue was dissolved in DCM, washed with a saturated bicarbonate solution, dried (in a phase separator cartridge), and concentrated under reduced pressure. The crude substance was purified by silica gel column chromatography and eluted with 80-100% siRNA in isohexane to obtain the marked compound as a white solid (1.76 g, 59%). 1 H NMR (400 MHz, CDCl3): δ 7.60-7.50 (m, 2H), 7.39 (d, 2H), 7.34-7.30 (m, 1H), 7.25 (t, 1H), 7.14 (d, 1H), 4.02-3.96 (m, 1H), 3.45 (s, 3H), 3.18-3.01 (m, 2H), 1.67 (s, 2H).

[0218] Intermediate 3 (2S)-4-(tert-butoxycarbonyl)-1,4-oxazepan-2-carboxylic acid [ka]

[0219] i) 3-{benzyl[(2S)-3-(benzyloxy)-2-hydroxypropyl]amino}propan-1-ol [ka] A solution of N-benzylpropanolamine (3.3 g) and benzyl(S)-(+)-glycidyl ether (3.6 g) in ethanol (40 mL) was heated at 40°C for 18 hours. The solvent was evaporated under reduced pressure to obtain the compound in question as a colorless oil (6.8 g, 100%), which was used without further purification. 1 H NMR (400 MHz, DMSO-d6): δ 7.29 (m, 10H), 4.54 (m, 1H), 4.45 (s, 2H), 4.36 (t, 2H), 3.76 (m, 1H), 3.44 (m, 5H), 2.47 (m, 4H), 1.57 (m, 2H).

[0220] ii) (2S)-4-benzyl-2-[(benzyloxy)methyl]-1,4-oxazepane [ka] At 0°C, 3-{benzyl[(2S)-3-(benzyloxy)-2-hydroxypropyl]amino}propan-1-ol (50.0 g, 0.153 mol) was stirred in THF (2.5 L), to which sodium hydride (15.2 g, 0.38 mol, 60% dispersion in oil) was added dropwise. The reaction was stirred at 0°C for 30 minutes, after which p-toluenesulfonylimidazole (37.8 g, 0.17 mol) was added dropwise. The reaction was heated to room temperature, stirred for 4 hours, and then cooled to 0°C. The reaction was quenched by carefully adding saturated sodium bicarbonate solution (70 mL). The solvent was removed under reduced pressure, and the crude residue was partitioned into water (400 mL) and HCl (400 mL). Each layer was separated, and the aqueous portion was extracted with HCl (400 mL x 2). The combined organic extracts were dried (with magnesium sulfate), filtered, and evaporated under reduced pressure to obtain oil. The oil was purified by silica gel column chromatography and eluted with a 0-50% siRNA gradient solution in isohexane to obtain the compound in the subtitle as a colorless oil (12.2 g, 26%). 1 H NMR (400 MHz, DMSO-d6): δ 7.36-7.20 (m, 10H), 4.45-4.35 (m, 2H), 3.81-3.65 (m, 2H), 3.60-3.39 (m, 2H), 3.42-3.31 (m, 2H), 3.25 (dd, 1H), 2.86 (d, 1H), 2.78-2.70 (m, 1H), 2.54-2.46 (m, 1H) 2.37 (dd, 1H), 1.89-1.77 (m, 1H), 1.78-1.66 (m, 1H).

[0221] iii) (2S)-2-(hydroxymethyl)-1,4-oxazepane-4-carboxylate tert-butyl [ka] Under nitrogen, (2S)-4-benzyl-2-[(benzyloxy)methyl]-1,4-oxazepane (12.2 g, 39.2 mmol) was dissolved in ethanol (250 mL), to which di-tert-butyl dicarbonate (10.22 g, 47.1 mmol) and 20% palladium-carbon (16.5 g) were added. The reaction mixture was shaken under a hydrogen atmosphere at 50 psi for 18 hours. The reaction mixture was then filtered through Celite and washed with methanol. The solvent was evaporated under reduced pressure to obtain the compound of the subtitle as a colorless oil (11.16 g). 1 H NMR (400 MHz, DMSO-d6): δ 4.72-4.66 (m, 1H), 4.00-3.89 (m, 1H), 3.80-3.61 (m, 1H), 3.60-3.47 (m, 2H), 3.49-3.21 (m, 4H), 3.07-2.88 (m, 1H), 1.79-1.69 (m, 2H), 1.40 (s, 9H).

[0222] iv)(2S)-4-(tert-butoxycarbonyl)-1,4-oxazepan-2-carboxylic acid At 0°C, (2S)-2-(hydroxymethyl)-1,4-oxazepane-4-carboxylate tert-butyl (13.2 g, 46.6 mmol) was dissolved in acetone (730 mL) and saturated sodium bicarbonate (218 mL), to which sodium bromide (1.46 g) and TEMPO (218 mg) were added. 1,3,5-trichloro-1,3,5-triazinan-2,4,6-trione (23.9 g, 102.5 mmol) was added dropwise, and the reaction mixture was heated to room temperature for 18 hours. Isopropanol (30 mL) was added to quench the reaction, and the mixture was stirred for 30 minutes. The reaction mixture was filtered through Celite and washed with ethyl acetate. The filtrate was evaporated under reduced pressure, dissolved in 100 mL of 1 M sodium carbonate solution, and extracted with ethyl acetate (200 mL x 2). The aqueous solution was acidified with 2M HCl (150 mL) and extracted with toluene (400 mL x 3). The combined organic extract was dried (with magnesium sulfate), filtered, and evaporated under reduced pressure to obtain the labeled compound as a colorless solid (7.86 g, 68%). 1H NMR (400 MHz, DMSO-d6): δ 12.71 (s, 1H), 4.22-4.15 (m, 1H), 3.98-3.80 (m, 2H), 3.70-3.50 (m, 2H), 3.45-3.11 (m, 1H), 3.21-3.06 (m, 1H), 1.71 (s, 2H), 1.40 (d, 9H).

[0223] Intermediate 3 (first alternative synthesis method) (2S)-4-(tert-butoxycarbonyl)-1,4-oxazepan-2-carboxylic acid i)(2S)-3-(dibenzylamino)-2-hydroxypropanoate methyl methyl (2S)-3-(dibenzylamino)-2-hydroxypropanoate [ka] (S)-oxirane-2-carboxylate methyl (117 g, 1134 mmol) and dibenzylamine (226 g, 1123 mmol) were heated overnight at 70°C under a nitrogen atmosphere. Furthermore, methyl (S)-oxirane-2-carboxylate (1.15 g, 11.2 mmol) was added. The mixture was then stirred at 80°C for 5 hours. Next, the mixture was left overnight at 50°C under reduced pressure (0-10 mbar). This yielded the desired product as a light brown, viscous oil (342.7 g, 1145 mmol). 1 Assay by 1H NMR: 89% w / w, effective yield 91%. 1 H NMR (400 MHz, CDCl3): δ 2.77 - 2.92 (m, 2H), 3.13 - 3.4 (s, broad, 1H), 3.49 (d, J=13.5, 2H), 3.63 (s, 3H), 3.74 (d, J=13.5, 2H), 4.21 (dd, J=4.3, 6.7, 1H), 7.18 - 7.34 (m, 10H).

[0224] ii) 3-{[(2S)-3-(dibenzylamino)-1-methoxy-1-oxopropan-2-yl]oxy}propa-2-enoic acid methyl [ka] (2S)-3-(dibenzylamino)-2-hydroxypropanoate methyl (342.7 g, 1018.8 mmol) was dissolved in toluene (200 mL). 4-methylmorpholine (22.4 mL, 203.8 mmol) was added, followed by the slow addition of methyl propiolate (108.8 g, 1273.6 mmol) over 60 minutes. Throughout this addition, the reaction temperature was maintained between 20 and 25°C by cooling in a water / ice bath. After stirring for 3 hours, the mixture was concentrated to obtain the desired product as a brown, viscous oil (447.6 g, 1167 mmol, Z / E isomer mixture). 1 1H NMR assay = 87% w / w (including both Z and E isomers). 1 H NMR (400 MHz, CDCl3): δ 2.9 - 3.02 (m, 2H), 3.53 (d, 2H), 3.64 (s, 3H), 3.66 (s, 2H), 3.70 (d, 2H), 4.41 (td, 1H), 4.86 (d, 0.08H), 5.20 (d, 0.92H), 6.33 (d, 0.08H), 7.16 - 7.34 (m, 11H), 7.43 (d, 0.92H). 13 C NMR (101 MHz, CDCl3): δ 51.13 (s), 52.30 (s), 54.64 (s), 58.92 (d, J = 5.6 Hz), 79.19 (s), 82.16 (s), 97.20 (s), 98.20 (s), 127.14 (s), 128.18 (d, J = 8.0 Hz), 128.88 (s), 138.54 (s), 138.88 (s), 156.76 (s), 161.01 (s), 167.59 (s), 169.04 (s).

[0225] iii)(2S)-3-amino-2-(3-methoxy-3-oxopropoxy)methyl propanoate [ka] Pd(OH)2 (20% charcoal, 50% water) (11.17 g, 79.50 mmol) was dried overnight under a nitrogen stream. Next, this was suspended in 1,4-dioxane (200 mL), and then added to a solution of 3-{[(2S)-3-(dibenzylamino)-1-methoxy-1-oxopropan-2-yl]oxy}propa-2-enoate methyl (438 g, 994 mmol) dissolved in 1,4-dioxane (3800 mL). The mixture was hydrogenated overnight at 30°C under a hydrogen pressure of 10 bar. The temperature was raised to 40°C, and the mixture was stirred for a further 2 days. The mixture was filtered and rinsed with dioxane (200 mL). Next, the dioxane solution (4527 g) was used directly in the next step. Assay = 4.6% w / w, effective yield 103%. 1 H NMR (400 MHz, CDCl3): δ 1.4 (s, 2H), 2.55 - 2.73 (m, 2H), 2.90 - 2.97 (dd, J=6.7, 13.5, 1H), 3.00 - 3.08 (dd, J=3.8, 13.5, 1H), 3.69 (s, 3H), 3.72 - 3.74 (m, 1H), 3.75 (s, 3H), 3.87 - 3.98 (ddd, 3.7, 6.3, 13.5, 2H).

[0226] iv)(2S)-5-oxo-1,4-oxazepan-2-carboxylate methyl [ka] Novozyme 435 (immobilized, 75 g) was added to a crude solution of (2S)-3-amino-2-(3-methoxy-3-oxopropoxy)propanoate methyl (204 g, 994 mmol) in dioxane (4.2 L). The mixture was stirred at 45°C for 2 days. Further Novozyme 435 (immobilized, 25 g) was added, and the mixture was stirred for a further 2 hours. The temperature was raised to 55°C, and the mixture was stirred for 24 hours. The mixture was filtered through a Celite filter, rinsed with MeOH, and then concentrated to obtain a soap-like solid (254 g). This was further purified by preparative HPLC to obtain 85.2 g (492 mmol) of the desired product as a colorless solid (>90% w / w). 1 (By 1H NMR). 1 H NMR (400 MHz, CDCl3): δ 6.98, (1H, s), 4.19 (2H, m), 3.77 (3H,s), 3.69 (1H,m), 3.59 (2H,m), 2.83 (1H, ddd) and 2.63 (1H, dd).

[0227] v)(2S)-5-oxo-1,4-oxazepan-2,4-dicarboxylic acid 4-tert-butyl,2-methyl [ka] Di-tert-butyl dicarbonate (192 g, 863.0 mmol) was added to a mixture of (2S)-5-oxo-1,4-oxazepane-2-carboxylate methyl (152.5 g, 863.0 mmol), N,N-dimethylpyridine-4-amine (2.11 g, 17.3 mmol), and THF (1200 mL). The resulting yellow suspension was then stirred at 30°C for 20 hours. Further di-tert-butyl dicarbonate (11.30 g, 51.8 mmol) was added, and the mixture was stirred at 30°C for another 20 hours. The mixture was almost concentrated to dryness on a 37°C water bath. MTBE (400 mL) was added, and the mixture was then almost concentrated to dryness. This procedure was repeated once more to remove the t-BuOH formed in the reaction. Finally, THF (300 mL) was added, and the mixture was concentrated to obtain a yellow oil, which was used directly in the next step. The termination is considered quantitative.1 H NMR (400 MHz, CDCl3): δ 1.48 (s, 9H); 2.77 (ddd, 1H, J = 16.1, 7.0, 1.9 Hz); 2.94 (ddd, 1H, J = 16.1, 9.3, 2.5 Hz); 3.75 (s, 3H); 3.80 (ddd, 1H, J = 12.9, 9.1, 2.0 Hz); 3.91 (dd, 1H, J = 16.0, 7.2 Hz); 4.12-4.30 (m, 2H); 4.38 (dd, 1H, J = 16.0, 1.4 Hz). 13 C NMR (126 MHz, CDCl3): δ 27.9, 42.3, 48.8, 52.6, 63.4, 77.5, 83.8, 152.1, 169.0, 172.6.

[0228] vi)(2S)-1,4-oxazepan-2,4-dicarboxylic acid 4-tert-butyl,2-methyl [ka] A crude mixture of (2S)-5-oxo-1,4-oxo-1,4-oxazepane-2,4-dicarboxylic acid 4-tert-butyl,2-methyl (212.4 g, 777.2 mmol) from the previous step in THF (2 L) was added with BH3-DMS solution (118 g, 1554 mmol) over 30 minutes. The reaction temperature was maintained between 20 and 23°C throughout this addition. The mixture was then stirred at 23°C for 17 hours. The mixture was slowly transferred to a MeOH solution (1.5 L). Next, the mixture was combined with the crude product obtained in a small-scale experiment (starting with 23.6 g of (2S)-5-oxo-1,4-oxazepane-2,4-dicarboxylic acid 4-tert-butyl,2-methyl, using the procedure described above). Next, the clear homogeneous solution was stirred at 20°C for 1 hour, then concentrated to almost dryness. MeOH (500 mL) was added, then concentrated to almost dryness, and this was repeated once more. Next, ACN (500 mL) was added, then concentrated to almost dryness, and this was repeated once more. Next, the crude product (24% w / w, 1The results were determined by 1H NMR, and the internal standard (benzyl benzoate) was stored as a solution in ACN (500 mL). Effective yield = 71%. 1 H NMR (400 MHz, MeOD, approximately 50:50 rotamer mixture): δ 1.51 (s, 9H); 1.84-1.93 (m, 2H); 3.20-3.34 (m, 1H); 3.42-3.56 (m, 1H); 3.70-3.81 (m, 5H); 4.02-4.12 (m, 2H); 4.36-4.41 (m, 1H). 13 C NMR (100.6 MHz, MeOD, approximately 50:50 rotamer mixture) δ 28.6, 31.0, 31.5, 47.8, 48.2, 51.0, 51.2, 52.6, 68.6, 68.7, 77.6, 77.8, 81.4, 81.6, 156.7, 156.9, 172.8, 172.9.

[0229] vii)(2S)-4-(tert-butoxycarbonyl)-1,4-oxazepan-2-carboxylic acid LiBr (375 g, 4319 mmol) was added to a mixture of ACN (700 mL), water (30 mL), TEA (187 g, 1851 mmol), and water (30 mL). Next, at a reaction temperature of 30 °C, (2S)-1,4-oxazepane-2,4-dicarboxylic acid 4-tert-butyl,2-methyl (160 g, 617 mmol) dissolved in ACN (200 mL) was added. The mixture was stirred vigorously overnight at 20 °C. Most of the ACN was removed by concentration. MTBE (500 mL) was added to the residue. The yellow aqueous layer was washed with MTBE (200 mL). Next, MTBE (400 mL) was added to the aqueous layer, and then it was acidified to approximately pH 2 using 2 M KHSO4. The aqueous layer was extracted with MTBE (300 mL x 2), the pooled organic layer was washed with water (100 mL), and then concentrated to obtain a colorless solid (170 g, 80% w / w). The solid was suspended in 30% MTBE (600 mL) in heptane, and the mixture was then stirred overnight. The mixture was filtered, the solid was washed with 25% MTBE in heptane (100 mL), and then dried under reduced pressure at 40°C. This yielded 140.1 g (571 mmol) of the desired product. 1 (93% w / w by 1H NMR, 99.7% ee by HPLC). 1 ¹H NMR (400 MHz, MeOD, mixture of two rotational isomers): δ 1.46 (s, 9H); 1.77-1.90 (m, 2H); 3.15-3.77 (m, 4H); 3.91-4.17 (m, 2H); 4.22-4.32 (m, 1H). 13 ¹³C NMR (100.6 MHz, MeOD, mixture of two rotational isomers): δ 28.5, 28.6, 31.0, 31.3, 47.8, 47.9, 51.4, 68.6, 69.0, 77.7, 78.0, 81.4, 81.7, 156.8, 157.0, 174.0, 174.2.

[0230] Intermediate 3 (2S)-4-(tert-butoxycarbonyl)-1,4-oxazepan-2-carboxylic acid (Second alternative synthesis method) i) 3-{benzyl[(2S)-3-(benzyloxy)-2-hydroxypropyl]amino}propan-1-ol [ka] 1219 g (7.16 mol) of the reactants 3-(benzylamino)propan-1-ol and 1200 g (7.16 mol) of (S)-2-((benzyloxy)methyl)oxirane were separately dissolved in 3 L each of 2-propanol, and then separately packed into inerted reactors and heated at 50°C for 24 hours. The reaction mixture was evaporated at 60°C at 110 mbar to obtain 2.48 kg of oil. The oil was dissolved in 1 L of toluene and evaporated to dryness. Yield: 2.45 kg, assay: approximately 95%, effective yield: approximately 98%. 1H NMR (400 MHz, CDCl3): δ 1.59 - 1.78 (m, 2H), 2.47 (dd, J=13.3, 1H), 2.53 - 2.65 (m, 2H), 2.71-2.78 (ddd, 5.6, 7.7, 13.2, 1H), 3.31 - 3.45 (m, 3H), 3.50 (d, 1H), 3.67 - 3.74 (m, J=13.3, 3H), 3.93-3.99 (ddt, J=4.1,4.1,6.2, 8.3, 1H), 4.48 (s, 2H), 7.18 - 7.36 (m, 10H).

[0231] ii) 3-{benzyl[(2S)-3-(benzyloxy)-2-hydroxypropyl]aminopropylmethanesulfonate [ka] 147 g (446 mmol) of the diol product from the previous experiment was dissolved in 400 mL of DCM and cooled to -1°C. 72.3 mL (446 mmol) of DIPEA was added to the reactor at -1°C. The solution was cooled to -6°C. Next, 51.1 g (446 mmol) of methanesulfonyl chloride from 200 mL of DCM was added dropwise to the diol solution at approximately -6°C to -2°C for 1 hour. After the addition, the mixture was stirred for 30 minutes and then poured onto 400 mL of ice. The phase was separated, washed twice with cold water, then washed twice with brine, and evaporated to obtain oil. The oil was diluted with DCM, extracted with aqueous sodium sulfate solution, filtered, and evaporated to obtain 176 g (97%) oil (85% assay). 1 H NMR (600 MHz, CDCl3): δ 1.82-1.87 (m, 2H), 2.47 - 2.56 (m, 3H), 2.59 - 2.67 (m, 1H), 2.86 (s, 3H), 3.02 (s, 1H), 3.38 - 3.45 (m, 2H), 3.49 (d,1H), 3.69 (d, 1H), 3.83 - 3.87 (m, 1H), 4.14-4.20 (m, 2H), 4.49 (s, 2H), 7.20 - 7.32 (m, 10H).

[0232] iii) (2S)-4-benzyl-2-[(benzyloxy)methyl]-1,4-oxazepane [ka] 169 g of the crude product from the previous experiment (approximately 85% of the assay, 143.65 g, 0.35 mol) was dissolved in 300 mL of dry THF and slowly added (over 5 hours) to 200 mL of dry THF containing NaH (1.4 equivalents, 18.46 g, 0.423 mol) under nitrogen at 25°C in a dry reactor. (Addition was started after washing the sodium hydride paste with heptane.) The reaction mixture was stirred overnight at 25°C. The next day, 400 mL of saturated aqueous bicarbonate solution was added to the reaction mixture at room temperature. Gas was released first. The phases were separated and the aqueous phase was discarded. The organic phase was evaporated to obtain oil. The oil was dissolved in 400 mL of isopropyl acetate. The isopropyl acetate solution was washed with 100 mL of 2 M NaOH (aq), then washed twice with water (100 mL), and washed with brine. The mixture was evaporated to obtain 136 g of product (65% w / w assay). Estimated yield: 88 g (81%) 0.28 mol. Chromatography: ¼ / heptane 254nm. Isolation yield: 81.6g (0.26mol, 74%) 1 H NMR (400 MHz, DMSO-d6): δ 1.66 - 1.76 (m, 1H), 1.77 - 1.87 (m, 1H), 2.37 (dd, 1H), 2.46 - 2.5 (m, 1H), 2.68 - 2.77 (m, 1H), 2.81 - 2.89 (m, 1H), 3.24 (dd, 1H), 3.37 (dd, 1H), 3.64 (d, 2H), 3.64 - 3.74 (m, 1H), 3.76 (ddd, 2H), 4.35 - 4.43 (m, 2H), 7.18 - 7.37 (m, 10H).

[0233] iv) (2S)-1,4-Oxazepan-2-ylmethanol [ka] 81.6 g (0.26 mol) of the product from the previous experiment was dissolved in 1 L of methanol and packed into a hydrogenation vessel under nitrogen. Catalyst PdOH2 (20%) (50% humidity) - 10 g of charcoal = 3 mol% was slurryed with ethanol and packed into a reaction vessel under nitrogen. The mixture was hydrogenated at 4.5 bar at ambient temperature for 72 hours. Approximately 50% was converted, 10g of new catalyst was added, the pressure was increased to 8 bar, and the temperature was raised from ambient temperature to 45°C. Hydrogenation was carried out overnight. Approximately 96% was converted. 3g of catalyst was added to the reaction mixture, and hydrogenation was continued for 6 hours. Complete conversion was achieved, the reaction mixture was filtered, and the sample was evaporated to obtain oil. 1 H NMR (500 MHz, MeOD): δ 1.60-1.79 (m, 2H), 2.42 - 2.53 (dd,J=8.8, 14, 1H), 2.62-2.81 (dddd, J=4.2,7.3,13.5,49,2H), 2.81-2.89 (dd, 1H), 2.94 (dd, 1H), 3.17 (s, 1H), 3.24 - 3.37 (qd, J=5.6, 11.4, 11.4, 11.4, 2H), 3.41-3.48 (m, 1H), 3.53 (td, J=3.9, 7.9, 7.8, 1H), 3.74-3.84 (dt, J=5.5, 5.5, 12.2, 1H).

[0234] v)(2S)-2-(hydroxymethyl)-1,4-oxazepane-4-carboxylate tert-butyl [ka] The methanol solution (approximately 1.2 L) of the product from the previous experiment (approximately 0.26 mol) was filtered off the catalyst and then treated with 54.3 g (0.25 mol) of anhydrous Boc under CO2 (g) at room temperature to initiate direct formation. The reaction was left overnight under nitrogen while stirring. The reaction mixture was evaporated to dryness to obtain 59 g (98%) of a light yellow liquid. 1 H NMR (500 MHz, MeOD): δ 1.47 (s, 9H), 1.81-1.93 (qt, J=3.51, 3.51, 6.3, 6.3, 6.3, 2H), 3.03-3.16 (ddd, J=9.5, 14.4, 21.8, 1H), 3.29-3.32 (dt, J=1.6, 1.6, 3.3, 1H), 3.32 - 3.41 (m, 1H), 3.43 - 3.56 (m, 3H), 3.56 - 3.71 (m, 2H), 3.78 (dd, 1H), 4.07 (tq, 1H).

[0235] vi)(2S)-4-(tert-butoxycarbonyl)-1,4-oxazepan-2-carboxylic acid 52.5 g of (2S)-2-(hydroxymethyl)-1,4-oxazepane-4-carboxylate tert-butyl (85% assay, 40.7 g) was dissolved in 300 mL of DCM. 0.5 g of TEMPO was dissolved in 100 mL of DCM. 3.88 g of tetrabutylammonium bisulfate was dissolved in 100 mL of DCM. These three DCM solutions were packed into a reaction vessel, and 100 mL of water was added. 350 mL of a 10-15% sodium hypochlorite solution was pH-adjusted to approximately 8-9 with approximately 100 mL of sodium bicarbonate (liquid + solid). 58 mL of a 0.5 M sodium bromide solution was added to the buffered solution. The resulting aqueous solution was added dropwise to a two-phase system consisting of a DCM mixture and water while stirring at 0°C. The reaction was exothermic. Following the addition, the color changed (from yellow to pale yellow). This color change indicates that the oxidant has been consumed. After 10 minutes, the jacket was set to -5°C and the internal temperature was maintained at approximately 10°C. After 45 minutes, the addition was completed and the reaction mixture was left overnight. Post-treatment: At room temperature, the off-white reaction mixture was pH-adjusted to approximately 2-3 with approximately 40 g of potassium bisulfate, the phases were separated, and the aqueous phase was washed with DCM (100 ml x 3). The obtained DCM (800 ml) solution was evaporated to obtain approximately 100 g of oil. This oil was dissolved in 400 ml of bicarbonate solution and extracted with DCM (75 ml x 2). The remaining aqueous phase was acidified to pH 2-3 with approximately 35-40 g of potassium bisulfate and extracted with DCM (75 ml x 5). The DCM was evaporated to obtain 40.7 g of white crystals; yield: 40.7 g, yield 85% based on the assay of the starting materials. The product contained 10% water. Purification: The product was slurryed with 200 mL of toluene and heated to 60°C until it became a solution. Approximately 100 mL of toluene was evaporated and removed, and the acidic product began to crystallize at 60°C. The mixture was cooled to room temperature. The product was filtered and washed with toluene. The product was dried under reduced pressure. 1 H NMR (600 MHz, CDCl3): δ 1.46 (s, 9H), 1.93 (s, 2H), 3.23 (ddt, 1H), 3.33 - 3.78 (m, 3H), 3.95 - 4.38 (m, 3H), 9.91(s,1H).

[0236] Intermediate 4 (2S)-2-{[(2S)-1-amino-3-(4-iodophenyl)-1-oxopropan-2-yl]carbamoyl}-1,4-oxazepan-4-carboxylate tert-butyl (2S)-4-(tert-butoxycarbonyl)-1,4-oxazepan-2-carboxylic acid (intermediate 3, 7.9 g, 32.2 mmol) and (S)-2-amino-3-(4-iodophenyl)propanamide (9.0 g, 32.2 mmol, prepared according to the procedure on page 45 of International Publication No. 2009 / 074829) were added to T3P (25 g, 39.3 mmol, 50% solution in DMF) (200 mL). TEA (25 mL, 180.3 mmol) was added, and the reaction was stirred at room temperature for 4 hours. The reaction mixture was then concentrated under reduced pressure. The resulting oil was dissolved in ELISA and washed sequentially with 2 M aqueous hydrochloric acid, saturated aqueous sodium bicarbonate, and sodium chloride solution. The organic extract was dried (with magnesium sulfate), filtered, and concentrated under reduced pressure to obtain the marked compound as a yellow foaming oil (13.1 g, 79%), which was used in the next step without further purification.

[0237] Intermediate 5 (2S)-2-{[(1S)-1-cyano-2-(4-iodophenyl)ethyl]carbamoyl}-1,4-oxazepane-4-carboxylate tert-butyl (2S)-2-{[(2S)-1-amino-3-(4-iodophenyl)-1-oxopropan-2-yl]carbamoyl}-1,4-oxazepan-4-carboxylate tert-butyl (intermediate 4, 8.86 g, 17.13 mmol) was dissolved in DCM (740 mL) and Burgess's reagent (8.16 g, 34.27 mmol) was added. The reaction mixture was stirred at room temperature for 24 hours, then transferred to a separatory funnel and washed with water. The organic extract was dried (in a phase separator cartridge) and concentrated under reduced pressure. The resulting solid was purified by silica gel column chromatography and eluted with 25% siRNA in isohexane to obtain a yellow oil. Tritulate with diethyl ether to obtain the marked compound as an off-white solid (6.05 g, 71%). 1H NMR (400 MHz, CDCl3): δ 7.66 (d, 2H), 6.98 (m, 3H), 5.06 (s, 1H), 4.22-3.92 (m, 3H), 3.70 (m, 0.5H), 3.54-3.20 (m, 2.5H), 3.09-2.89 (m, 3H), 1.88 (s, 2H), 1.42 (s, 9H).

[0238] Intermediate 6 (2S)-2-[[(1S)-2-amino-2-oxo-1-[[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]methyl]ethyl]carbamoyl]-1,4-oxazepane-4-carboxylate tert-butyl Under nitrogen, (2S)-2-{[(2S)-1-amino-3-(4-iodophenyl)-1-oxopropan-2-yl]carbamoyl}-1,4-oxazepan-4-carboxylate tert-butyl (intermediate 4, 0.5 g, 0.97 mmol) was stirred in dry DMSO (2.5 mL) to which Pin2B2 (0.32 g, 1.26 mmol), potassium acetate (0.28 g, 2.9 mmol), and Pd(dppf)Cl2·DCM (0.039 g, 5 mol%) were added. The reaction was heated at 85 °C for 5 hours and left overnight at room temperature. Water (15 mL) was added, and the mixture was extracted with ELISA (50 mL × 2). The combined extract was washed with saturated sodium chloride (20 mL), dried (magnesium sulfate), and evaporated under reduced pressure. The resulting oil was purified by silica gel column chromatography and eluted with toluene to obtain the labeled compound as a colorless oil (0.3 g, 60%). 1¹H NMR (400 MHz, CDCl3): δ 7.75 (d, 2H), 7.28-7.21 (m, 2H), 5.30 (s, 1H), 4.60 (m, 1H), 4.18-3.98 (m, 2H), 3.51-3.42 (m, 1H), 3.12 (t, 2H), 2.80 (s, 1H), 2.05 (s, 2H), 1.88 (s, 1H), 1.60 (s, 4H), 1.54-1.33 (m, 6H), 1.40-1.16 (m, 12H) (3 exchangeable protons were not observed).

[0239] Examples

[0240] Example 1 (2S)-N-[(1S)-1-cyano-2-(4'-cyanobiphenyl-4-yl)ethyl]-1,4-oxazepan-2-carboxamide [ka] i) (2S)-2-{[(1S)-1-cyano-2-(4'-cyanobiphenyl-4-yl)ethyl]carbamoyl}-1,4-oxazepan-4-carboxylate tert-butyl (2S)-4-(tert-butoxycarbonyl)-1,4-oxazepane-2-carboxylic acid (intermediate 3, 0.294 g, 1.2 mmol) was dissolved in DCM (15 mL) and 2-pyridinol-1-oxide (0.155 g, 1.4 mmol), TEA (0.36 g, 3.6 mmol), and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (0.268 g, 1.4 mmol) were added. After 20 minutes, 4'-[(2S)-2-amino-2-cyanoethyl]biphenyl-4-carbonitrile (intermediate 1, 0.296 g, 1.2 mmol) was added, and the mixture was stirred for 3 hours and left at room temperature for 18 hours. The mixture was heated at 40°C for 4 hours, and then water (15 mL) was added. After 10 minutes, the DCM was dried (in the phase separator cartridge) and evaporated under reduced pressure. The resulting yellow oil was purified by silica gel column chromatography to obtain the compound indicated in the subtitle (0.29 g, 52%). It was used in the next step without further purification.

[0241] ii) (2S)-N-[(1S)-1-cyano-2-(4'-cyanobiphenyl-4-yl)ethyl]-1,4-oxazepan-2-carboxamide Using (2S)-2-{[(1S)-1-cyano-2-(4'-cyanobiphenyl-4-yl)ethyl]carbamoyl}-1,4-oxazepane-4-carboxylate tert-butyl, the preparation was carried out according to step ii) of Method A, and the marked compound was obtained as a white solid (60 mg, 28%). 1 ¹H NMR (400 MHz, CDCl3): δ 7.77-7.65 (m, 4H), 7.62-7.57 (m, 2H), 7.40 (d, 2H), 7.11 (d, 1H), 5.18-5.11 (m, 1H), 4.19-4.14 (m, 1H), 4.06-3.96 (m, 2H), 3.75-3.69 (m, 1H), 3.56-3.48 (m, 2H), 3.18-3.05 (m, 3H), 2.95-2.90 (m, 1H), 2.70 (ddd, 1H) (one exchangeable proton was not observed). LCMS (10cm_ESCI_Formic_MeCN) t R 2.57 (mim) m / z 375 (MH + ).

[0242] Example 2 (2S)-N-{(1S)-1-cyano-2-[4-(3-methyl-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl)phenyl]ethyl}-1,4-oxazepan-2-carboxamide [ka]

[0243] i) (2S)-2-({(1S)-1-cyano-2-[4-(3-methyl-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl)phenyl]ethyl}carbamoyl)-1,4-oxazepane-4-carboxylate tert-butyl [ka] (2S)-4-(tert-butoxycarbonyl)-1,4-oxazepan-2-carboxylic acid (intermediate 3, 490 mg, 2.0 mmol) was dissolved in DCM (15 mL) and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (468 mg, 2.44 mmol) and 2-pyridinol 1-oxide (271 mg, 2.44 mmol) were added. The reaction was stirred at room temperature for 30 minutes, after which (2S)-2-amino-3-[4-(3-methyl-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl)phenyl]propannitrile (intermediate 2, 586 mg, 2.0 mmol) and DiPEA (1.79 mL, 10 mmol) were added. The reaction was stirred at room temperature for 18 hours, and then transferred to a separatory funnel. The mixture was washed with 2M hydrochloric acid, a saturated sodium bicarbonate solution, and brine. The organic extract was passed through a hydrophobic frit / phase separator and concentrated under reduced pressure. The crude substance was purified by silica gel column chromatography and eluted with 0-60% siRNA in isohexane to obtain the subtitle compound as oil (457 mg, 44%). 1 H NMR (400 MHz, CDCl3): δ 7.63-7.52 (m, 2H), 7.38 (d, 2H), 7.36-7.24 (m, 2H), 7.35-6.98 (m, 2H), 5.18 (t, 1H), 4.22-3.97 (m, 2H), 3.76-3.67 (m, 0.5H), 4.10-2.94 (m, 4.5H), 3.35-3.26 (m, 1H), 3.24-3.04 (m, 3H), 2.06-1.82 (m, 2H), 1.47 (s, 10H).

[0244] ii) (2S)-N-{(1S)-1-cyano-2-[4-(3-methyl-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl)phenyl]ethyl}-1,4-oxazepan-2-carboxamide (2S)-2-({(1S)-1-cyano-2-[4-(3-methyl-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl)phenyl]ethyl}carbamoyl)-1,4-oxazepane-4-carboxylate tert-butyl (457 mg, 0.85 mmol) was dissolved in formic acid (3 mL) and heated at 50°C for 10 minutes on a preheated hot plate stirrer. The reaction was then concentrated under reduced pressure, dissolved in DCM, and washed with saturated sodium bicarbonate solution. The organic extract was passed through a hydrophobic frit / phase separator and concentrated under reduced pressure. The resulting foam was purified by silica gel column chromatography and eluted with 0-5% methanolic ammonia (7N) in DCM to obtain the marked compound as a solid (230 mg, 64%). 1 ¹H NMR (400 MHz, CDCl3): δ 7.59-7.51 (m, 2H), 7.39 (dd, 2H), 7.33-7.23 (m, 3H), 7.14 (d, 1H), 5.23-5.12 (m, 1H), 4.12-4.06 (m, 1H), 4.05-3.95 (m, 1H), 3.81-3.71 (m, 1H), 3.46 (s, 3H), 3.34-3.26 (m, 1H), 3.19-3.00 (m, 3H), 2.99-2.82 (m, 2H), 1.92-1.77 (m, 2H) (one exchangeable proton was not observed). LCMS (10cm_ESCI_Formic_MeCN) t R 2.48 (min) m / z 375 (MH + ).

[0245] Example 2 (Alternative Synthesis Method) (2S)-N-{(1S)-1-cyano-2-[4-(3-methyl-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl)phenyl]ethyl}-1,4-oxazepan-2-carboxamide i) 5-Chloro-1,3-benzoxazole-2(3H)-one [ka] CDI (497 g, 3.07 mol) was added to a solution of 2-amino-4-chlorophenol (400 g, 2.79 mol) in 2-MeTHF (6 L) under N2 (exothermic, 11.0°C to 22.0°C). The reaction mixture was heated under reflux for 1 hour. The mixture was cooled to room temperature and washed with 2 M HCl (aq) (6 L), 8% NaHCO3 (aq) (6 L), and brine (3 L). The organic layer was dried over MgSO4, filtered, and evaporated. The product was obtained as a light brown solid (456.1 g, yield 97%, LC purity >99%). 1 H NMR (270 MHz, DMSO-d6): δ 12.0-11.5 (brs, 1H), 7.31 (d, 1H), 7.12 (m, 2H). LCMS (5cm_ESCI, aq. formic acid_metanol) t R 3.87 (mim) m / z 169.8 (MH + ).

[0246] ii) 5-Chloro-3-methyl-1,3-benzoxazole-2(3H)-one [ka] Cs2CO3 (2136.4g, 6.56mol) was added to a solution of 5-chloro-1,3-benzoxazole-2(3H)-one (step i) (1111.8g, 6.56mol) in DMF (4.12L) while maintaining the temperature at 0-5°C. Next, MeI (450ml, 7.21mol) was slowly added while maintaining the temperature at 0-5°C. The reaction mixture was heated to room temperature and stirred overnight. The mixture was cooled to 0-5°C and H2O (4.12L) was slowly added. Next, the reaction mixture was heated to room temperature and stirred for 15 minutes. The solid was filtered and washed with water (980ml x 4). The filtered cake was vacuum-dried overnight at 55°C (1149.9g, yield 96%, LC purity >99%, H2O: (Karl Fischer) 0.1%). 1H NMR (270 MHz, DMSO-d6): δ 7.45 (d, 1H), 7.35 (d, 1H), 7.15 (dd, 1H), 3.35 (s, 3H). LCMS (5cm_ ESCI_aq. formic acid_methanol) t R 4.13 (mim) m / z 183.8 (M + ).

[0247] iii) 3-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3-benzoxazole-2(3H)-one [ka] A solution of 5-chloro-3-methyl-1,3-benzoxazole-2(3H)-one (step ii) (350 g, 1.91 mol), B2pin2 (581.0 g, 2.29 mol), and KOAc (561.3 g, 5.72 mol) was degassed under vacuum and purged with N2 (×3). Pd(OAc)2 (12.9 g, 57.2 mmol) and XPhos (54.6 g, 114 mmol) were added, the mixture was degassed under vacuum, and purged with N2 (×3). The mixture was heated to 75°C. Significant exothermic reaction was observed at approximately 70°C, which caused the mixture to heat to reflux (100°C). The reaction mixture was stirred for 1 hour without heating. HPLC analysis showed that 2.5% of the starting material remained, so the mixture was heated to 85°C for 1 hour. No further changes were observed at this stage. Furthermore, B2pin2 (14.6 g, 57.2 mmol), KOAc (5.7 g, 57.2 mmol), Pd(OAc)2 (12.9 g, 57.2 mmol), and XPhos (27.3 g, 57.2 mmol) were added, and the mixture was stirred at 75°C for 1 hour. HPLC analysis showed that no starting material remained. The mixture was cooled to room temperature, filtered through a Celite (501 g) pad, and the cake was washed with SiO2 (2240 ​​ml). The filtrate was combined with two other batches (350 g x 2) prepared in the same manner and evaporated. This yielded 1865.1 g of product as a gray solid (yield 97%, purity 90.0% by LC). 1 Purity of 82±2% was determined by 1H NMR (DMSO-d6) assay versus TCNB. 1H NMR (270MHz, DMSO-d6): δ 7.40-7.50 (m, 2H), 7.30 (d, 1H), 3.40 (s, 3H), 1.30 (s, 12H). LCMS (5cm_ ESCI_aq. formic acid_methanol_) t R 4.91 (mim) m / z 276.1 (MH + ).

[0248] iv) Nα-(tert-butoxycarbonyl)-4-(3-methyl-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl)-L-phenylalaninamide [ka] 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3-benzoxazole-2(3H)-one (step iii)) (859 g, 700 g active, 2.544 mol) and (S)-1-carbamoyl-2-(4-iodophenyl)ethylcarbamate tert-butyl (prepared according to the procedure on page 47 of International Publication No. 2009 / 074829) (903 g, 2.313 mol) were added to a suspension of these compounds in dioxane (4.1 L) with 2 M K2CO3 (2.3 L). The suspension was degassed under vacuum and purged with N2 (×3). Pd(dppf)Cl2·DCM (28.33 g, 0.0347 mol) was added, and the reaction mixture was heated at 75 °C for 3 hours. The mixture was cooled to room temperature and diluted with water (6.4 L). The suspension was stirred overnight at room temperature; the solid was filtered and washed with water (1 L x 3). The product was dried at 45°C for 3 days (1269.1 g, 1 Yield 133% by 1H NMR - contains pinacol-related impurities and dioxane, LC purity 94.3%, H2O: (Karl Fischer) 3.35%. 1 H NMR (270 MHz, DMSO-d6): δ 7.62-7.34 (m, 7H), 7.04 (brs, 2H), 6.86 (d, 1H) 4.12 (m, 1H), 3.40 (s, 3H), 3.00 (dd, 1H), 2.78 (dd, 1H), 1.30 (s, 9H). LCMS (5cm_ESI_Water_MeCN) t R 4.51 (mim) m / z 312 (MH + ).

[0249] v)4-(3-methyl-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl)-L-phenylalaninamide [ka] Under N2 conditions, a very concentrated suspension of Nα-(tert-butoxycarbonyl)-4-(3-methyl-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl)-L-phenylalaninamide (step iv) (1269 g, step iv) (952 g, 2.3138 mol) in DCM (2.1 L) was added dropwise over 1 hour while maintaining the temperature at approximately 15°C (the suspension became more fluid after the addition of approximately 0.5 L of 4.1 M HCl in dioxane). After 2 hours, the mixture was diluted with water (5.6 L) and stirred at room temperature for 30 minutes. The mixture was then filtered through a Celite (500 g) pad to remove any undissolved substances - very slow filtration; the Celite was checked for the product by LC. The pad was washed with water (400 ml). The layers were separated into DCM / dioxane-water. The aqueous layer was cooled to approximately 5°C, and 35% NH3(aq) (700 ml) was slowly added until the pH reached 9-10. The suspension was stirred overnight, and then the product was filtered and washed with water (400 ml x 3). The product was dried under vacuum at 45°C (off-white solid, 489.4 g, yield 68% over two steps, purity 99.4% by LC, >99% EP). 1 ¹H NMR assay vs. TCNB in ​​DMSO: purity 98±2%, H₂O: (Karl Fischer) 0.92%. 1 H NMR (270 MHz, DMSO-d6): δ 7.59-7.30 (m, 7H), 6.98 (brs, 1H), 3.36 (m, 4H), 2.95 (dd, 1H), 2.67 (dd, 1H) 1.86 (brs, 2H). LCMS (5cm_ESI_Water_MeCN) t R 2.76 (mim) m / z 312 (MH + ).

[0250] vi)(2S)-2-({(1S)-1-cyano-2-[4-(3-methyl-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl)phenyl]ethyl}carbamoyl)-1,4-oxazepane-4-carboxylate tert-butyl [ka] Under N2 conditions, 4-(3-methyl-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl)-L-phenylalaninamide (step v)) (756 g, active 733 g, 2.354 mol) and (2S)-4-(tert-butoxycarbonyl)-1,4-oxazepan-2-carboxylic acid (intermediate 3) were dissolved in DMF (3 L) and DiPEA (1230 ml, 7.062 mol) was added. While maintaining the temperature at <25°C, T3P (50% w / w, 1924 ml, 3.296 mol) was added dropwise to DMF over 1.5 hours. After 30 minutes, the LC completion check indicated completion of the coupling reaction. Next, DiPEA (1230 ml, 7.062 mol) was added and the reaction mixture was heated to 50°C. T3P (50% w / w, 3986 ml, 6.827 mol) was added dropwise to DMF over 1 hour (no exothermic reaction was observed). The reaction mixture was stirred at 50°C for 4 hours, then stirred overnight at room temperature. The mixture was cooled to 10°C and diluted with 2-MeTHF (4 L) and water (5.6 L, exothermic). Each layer was separated, and the aqueous layer was extracted with 2-MeTHF (4 L x 2). The combined organic extract was dried over MgSO4, filtered, and concentrated under reduced pressure. The product was obtained as a light brown solid in 98% yield (1242 g (activity 1205 g), corrected yield 98%, LC purity 98.4%). 1 1H NMR assay vs. TCNB 97±2%, 1 Main impurities detected by 1H NMR: 2-MeTHF 1.9%, DMF 0.6%.

[0251] vii)(2S)-N-{(1S)-1-cyano-2-[4-(3-methyl-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl)phenyl]ethyl}-1,4-oxazepan-2-carboxamide (2S)-2-({(1S)-1-cyano-2-[4-(3-methyl-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl)phenyl]ethyl}carbamoyl)-1,4-oxazepane-4-carboxylate tert-butyl (step vi)) (1776 g, 1671 g active, 3.210 mol) was mixed in formic acid / water (4.2 L / 440 ml) and stirred over a buchi under reduced pressure (300-500 mbar) at 35-37°C. After 3 hours, the LCMS completion check showed 93.95% product and 0.5% starting material. The mixture was concentrated (4 hours) to obtain an oily residue. The residue was dissolved in water (4.4 L) and washed with TBME (2.2 L). The aqueous layer was vigorously stirred and treated with NH3(aq) (1.8 L) at <25°C until the pH reached 9-10. The mixture was stirred at room temperature for 3 hours. The solid was filtered and washed with water (1 L x 3). The filtered cake was dried overnight at 45°C. The product was then obtained as a light brown solid (1498 g, activity 1333 g, LC 91.5%). 1 ¹H NMR assay vs. TCNB 89±2%, H₂O:(Carl Fischer) 4.63%).

[0252] The crude product was recrystallized from EtOH / H2O in two batches (747g x 2). Batch A: The crude product (747g) was dissolved in EtOH (8L) under reflux under N2. Water (1.6L) was slowly added. The mixture was hot filtered (65°C) to remove black particles (filtrate temperature 50°C), and then stirred overnight at 40°C. The suspension was cooled to 10°C over 4 hours and maintained at this temperature for 3 hours. The product was filtered and washed with EtOH / H2O (8:2, 500ml x 3), and then washed with water (500ml x 3). The filtered cake was dried overnight at 45°C (473g, purity 97.7% by LC, Pd level 71.4ppm). From batch B, 436 g of product was obtained (purity 95.8% by LC, Pd level 65.8 ppm). The liquids from both batches were combined and concentrated to approximately 8 L. The liquid was left at room temperature overnight. The solid was filtered, washed with EtOH / H2O (8:2, 400 ml x 3), and then washed with water (400 ml x 3). The product was dried overnight at 45°C. This yielded an additional 88 g of product (LC purity 95.0%).

[0253] These products (LC purity of the blend 95.69%) were recrystallized from EtOH / H2O in two batches (Batch C: 520g, Batch D: 520g). Under reflux in batch C:N2, the crude product (520g) was dissolved in EtOH (6.24L). Water (1248ml) was slowly added. The mixture was cooled to 40°C (3 hours), 0.5g of the labeled compound was seeded, and the mixture was stirred at 40°C for 10 hours. Next, the mixture was cooled to 26°C over 7 hours. The resulting suspension was cooled to 10°C and stirred at that temperature for 6 hours. The product was filtered and washed with EtOH / water (8:2, 500ml x 3) and water (500ml x 3). The filtered cake was dried at 45°C for 2 days. The product was obtained as a gray solid (418g, yield approximately 56%, LCMS purity 97.5%, chiral LC 100%). 1 1H NMR (DMSO-d6) assay vs. TCNB (100±2%). Batch D: 418g, yield approximately 56%, LC / MS purity 97.5%, chiral LC 100%. 1 1H NMR (DMSO-d6) assay vs. TCNB 100±2%

[0254] These products were blended with substances obtained from intermediate-scale reactions carried out using the same method and re-analyzed (968g, LC purity 98.04%, chiral LC 100%). 1 ¹H NMR assay vs. TCNB 99±2%, 1 ¹H NMR revealed 0.35% EtOH, H₂O (Karl Fischer) 4.58%, Pd 57.6 ppm, and XRPD (powder X-ray diffraction) morphology A.

[0255] [Table 2] [Table 3]

[0256] Example 2: Preparation of Crystal Form B (2S)-N-{(1S)-1-cyano-2-[4-(3-methyl-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl)phenyl]ethyl}-1,4-oxazepan-2-carboxamide, Form A (5 g), prepared by the method described above, was packed into a reaction vessel. Acetone (35 ml) was added, and the mixture was heated in a heating block at 60-65°C. The heating block was switched off, and the resulting solution was allowed to cool to room temperature. The resulting suspension was filtered, and the filtrate was dried overnight in a vacuum oven at 40°C and ≤600 mbar. XRPD (powder X-ray diffraction), Form B.

[0257] [Table 4] [Table 5]

[0258] Example 2: Preparation of crystalline form C (2S)-N-{(1S)-1-cyano-2-[4-(3-methyl-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl)phenyl]ethyl}-1,4-oxazepan-2-carboxamide, form A (50 mg), prepared by the method described above, was packed into a 1.5 mL scintillation vial. Propan-2-ol (1 ml) was added, and the mixture was placed in an orbital shaker equipped with a heating block at 40°C for 1 day. The resulting suspension was filtered, and the filtrate was dried. XRPD (powder X-ray diffraction), form C.

[0259] [Table 6] [Table 7]

[0260] Example 2: Preparation of xinafoate, crystalline form A (2S)-N-{(1S)-1-cyano-2-[4-(3-methyl-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl)phenyl]ethyl}-1,4-oxazepan-2-carboxamide, form A (100 mg), prepared by the method described above, was packed into a 1.5 mL scintillation vial. Approximately 48 mg of 1-hydroxy-2-naphthoic acid was added. Subsequently, 1.5 mL of ACN and 0.03 mL of water were added, and the mixture was stirred at room temperature for approximately 6 hours using a magnetic stirring rod. The vial was kept closed during this stirring. The resulting suspension was centrifuged at 7500 rpm for 5 minutes, and the supernatant was removed using a Pasteur pipette. The moist solid residue was dried in a vacuum oven at 30°C and 30 mbar for approximately 60 hours. XRPD (powder X-ray diffraction), xinafoate of form A.

[0261] [Table 8] [Table 9]

[0262] Example 2: Preparation of R-mandelate, crystalline form A (2S)-N-{(1S)-1-cyano-2-[4-(3-methyl-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl)phenyl]ethyl}-1,4-oxazepan-2-carboxamide, form A (120 mg), prepared by the method described above, was packed into a 1.5 mL scintillation vial. Approximately 45 mg of R-(-)-mandelic acid was added. Subsequently, 1.5 mL of ACN and 0.04 mL of water were added, and the mixture was stirred at room temperature for approximately 6 hours using a magnetic stirring rod. The vial was kept closed during this stirring. The resulting suspension was centrifuged at 7500 rpm for 5 minutes, and the supernatant was removed using a Pasteur pipette. The moist solid residue was dried in a vacuum oven at 30°C and 30 mbar for approximately 60 hours. XRPD (powder X-ray diffraction), R-mandelic acid of form A.

[0263] [Table 10] [Table 11]

[0264] Example 3 (Method A) (2S)-N-{(1S)-1-cyano-2-[4-(3,7-dimethyl-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl)phenyl]ethyl}-1,4-oxazepan-2-carboxamide [ka] i) (2S)-2-({(1S)-1-cyano-2-[4-(3,7-dimethyl-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl)phenyl]ethyl}carbamoyl)-1,4-oxazepane-4-carboxylate tert-butyl 5-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-3,7-dimethyl-1,3-benzoxazole-2(3H)-one (boronic acid ester 1, 154 mg, 0.56 mmol) and (2S)-2-{[(1S)-1-cyano-2-(4-iodophenyl)ethyl]carbamoyl}-1,4-oxazepan-4-carboxylate tert-butyl (intermediate 5, 266 mg, 0.53 mmol) were dissolved in ACN (13 mL) and water (0.5 mL). Potassium carbonate (110 mg, 0.80 mmol) was added, and the reaction mixture was degassed for 20 minutes, after which Pd(dppf)Cl2·DCM (43 mg, 0.053 mmol) was added. The reaction mixture was heated at 80°C for 90 minutes. Subsequently, the reaction was concentrated under reduced pressure, purified by silica gel column chromatography, and eluted with a 0-80% siRNA gradient solution in isohexane to obtain the compound of the subtitle as a light brown solid (242 mg, 85%). 1 H NMR (400 MHz, CDCl3): δ 7.55 (d, 2H), 7.36 (d, 2H), 7.16-7.02 (m, 3H), 6.96 (s, 1H), 5.16 (s, 1H), 4.17-4.00 (m, 3H), 3.56-3.48 (m, 1H), 3.53-3.36 (m, 3H), 3.21-3.12 (m, 2H), 2.44 (s, 3H), 1.95 (d, 2H), 1.47 (s, 9H), 0.94-0.87 (m, 2H).

[0265] ii) (2S)-N-{(1S)-1-cyano-2-[4-(3,7-dimethyl-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl)phenyl]ethyl}-1,4-oxazepan-2-carboxamide (2S)-2-({(1S)-1-cyano-2-[4-(3,7-dimethyl-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl)phenyl]ethyl}carbamoyl)-1,4-oxazepane-4-carboxylate tert-butyl (240 mg, 0.45 mmol) was dissolved in formic acid (3 mL) and heated at 50°C for 10 minutes on a preheated hot plate stirrer. The reaction was then concentrated under reduced pressure, dissolved in DCM, and washed with saturated sodium bicarbonate solution. The organic extract was dried (in a phase separator cartridge) and concentrated under reduced pressure. The solid was purified by silica gel column chromatography and eluted with 0-2% methanolic ammonia (7N) in DCM to obtain the marked compound as a white solid (54 mg, 27%). 1 ¹H NMR (400 MHz, DMSO-d6): δ 8.62 (d, 1H), 7.65 (d, 2H), 7.38 (d, 3H), 7.28 (s, 1H), 5.03 (q, 1H), 4.00 (dd, 1H), 3.90-3.82 (m, 1H), 3.73 (ddd, 1H), 3.39 (s, 3H), 3.32 (s, 3H), 3.24-3.13 (m, 2H), 3.04 (dd, 1H), 2.82-2.74 (m, 1H), 2.38 (s, 2H), 1.80-1.68 (m, 2H) (one exchangeable proton was not observed). LCMS (10cm_ESCI_Formic_MeCN) t R 2.58 (mim) m / z 435 (MH + ).

[0266] Example 4 (Method B) 4'-[(2S)-2-cyano-2-{[(2S)-1,4-oxazepan-2-ylcarbonyl]amino}ethyl]biphenyl-3-ylmethanesulfonate [ka] i) (2S)-2-{[(2S)-1-amino-3-{3'-[(methylsulfonyl)oxy]biphenyl-4-yl}-1-oxopropan-2-yl]carbamoyl}-1,4-oxazepan-4-carboxylate tert-butyl (2S)-2-[[(1S)-2-amino-2-oxo-1-[[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]methyl]ethyl]carbamoyl]-1,4-oxazepan-4-carboxylate tert-butyl, 2-({(2S)-1-amino-1-oxo-3-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2 A suspension of (30 mL) tert-butyl (intermediate 6, 0.21 g, 0.4 mmol), (3-iodophenyl)methanesulfonate (0.13 g, 0.44 mmol), and potassium carbonate (0.16 g, 1.2 mmol) in ACN (30 mL) and water (1.2 mL) was degassed under nitrogen for 10 minutes. Pd(dppf)Cl2·DCM complex (0.032 g, 10 mol%) was added, and the reaction mixture was heated at 80 °C for 120 minutes. The solvent was removed under reduced pressure, and the residue was treated with water (20 mL) and DCM (25 mL). The DCM was dried (in a phase separation cartridge) and evaporated under reduced pressure to obtain the subtitle compound as dark brown glass (0.24 g, >100%). 1 ¹H NMR (400 MHz, CDCl3): δ 7.54-7.43 (m, 5H), 7.35-7.19 (m, 3H), 5.58 (m, 1H), 4.71 (s, 1H), 4.21-3.94 (m, 3H), 3.81-3.76 (m, 1H), 3.52-3.44 (m, 3H), 3.23-3.14 (m, 4H), 2.80 (s, 1H), 2.20-1.54 (m, 1H), 1.45 (s, 9H) (3 exchangeable protons were not observed).

[0267] ii) (2S)-2-{[(1S)-1-cyano-2-{3'-[(methylsulfonyl)oxy]biphenyl-4-yl}ethyl]carbamoyl}-1,4-oxazepane-4-carboxylate tert-butyl (2S)-2-{[(2S)-1-amino-3-{3'-[(methylsulfonyl)oxy]biphenyl-4-yl}-1-oxopropan-2-yl]carbamoyl}-1,4-oxazepane-4-carboxylic acid tert-butyl (0.24 g) was stirred in DCM (20 mL) and Burgess's reagent (0.11 g, 0.046 mmol) was added. After 3 days, an additional reagent (0.11 g, 0.046 mmol) was added and stirring was continued for 6 hours. After letting the reaction stand overnight, the mixture was washed with water (20 mL). The organic extract was dried (in a phase separation cartridge) and evaporated under reduced pressure. The residue was purified by silica gel column chromatography and eluted with 0-100% siRNA in isohexane to obtain the subtitle compound as a colorless glass (0.18 g, 83% over two steps). 1 ¹H NMR (400 MHz, CDCl3): δ 7.61-7.45 (m, 5H), 7.38 (m, 3H), 7.06 (s, 1H), 5.17 (s, 1H), 4.20-3.99 (m, 2H), 3.75-3.63 (m, 1H), 3.57-3.37 (m, 3H), 3.49-2.85 (m, 3H), 1.94 (s, 2H), 1.57 (s, 1H), 1.51-1.35 (m, 9H), 1.33 (s, 1H) (one exchangeable proton was not observed).

[0268] iii) 4'-[(2S)-2-cyano-2-{[(2S)-1,4-oxazepan-2-ylcarbonyl]amino}ethyl]biphenyl-3-ylmethanesulfonate A solution of (2S)-2-{[(1S)-1-cyano-2-{3'-[(methylsulfonyl)oxy]biphenyl-4-yl}ethyl]carbamoyl}-1,4-oxazepane-4-carboxylate tert-butyl (0.18 g, 0.33 mmol) in formic acid (3 mL) was heated at 50°C for 15 minutes. The mixture was evaporated under reduced pressure. The residue was dissolved in DCM (20 mL) and stirred with saturated sodium bicarbonate (30 mL). Each layer was separated, the organic extracts were dried (in a phase separation cartridge), and evaporated under reduced pressure. The residue was purified by silica gel column chromatography and eluted with 2% 7N methanolic ammonia in DCM. The resulting solid was recrystallized from a 1:1 diisopropyl ether:siRNA to obtain the marked compound as a colorless solid (50 mg, 34%). 1 ¹H NMR (400 MHz, CDCl3): δ 7.61-7.46 (m, 5H), 7.40 (dd, 2H), 7.38-7.18 (m, 1H), 7.18 (d, 1H), 5.23-5.12 (m, 1H), 4.12-4.06 (m, 1H), 4.05-3.95 (m, 1H), 3.81-3.71 (m, 1H), 3.35-3.26 (m, 1H), 3.22-3.09 (m, 4H), 3.07-2.81 (m, 3H), 1.91-1.77 (m, 2H) (two exchangeable protons were not observed). LCMS (10cm_ESCI_Bicarb_MeCN) t R 2.75 (min) m / z 444 (MHz) + ).

[0269] Examples 5-33 Using the above method and intermediates, the following compounds were prepared: [ka] [Table 12-1] [Table 12-2] Table 12-3 Table 12-4 Table 12-5 Table 12-6 Table 12-7 Table 12-8 Table 12-9 Table 12-10 Table 12-11 Table 12-12 Table 12-13 Table 12-14 Table 12-15

[0270] Example 34 (2S)-N-[(1S)-1-cyano-2-(4'-cyanobiphenyl-4-yl)ethyl]-1,4-oxazepan-2-carboxamide and (2R)-N-[(1S)-1-cyano-2-(4'-cyanobiphenyl-4-yl)ethyl]-1,4-oxazepan-2-carboxamide diastereomer mixture [ka] i) 2-{[(1S)-1-cyano-2-(4'-cyanobiphenyl-4-yl)ethyl]carbamoyl}-1,4-oxazepan-4-carboxylate tert-butyl rac-4-(tert-butoxycarbonyl)-1,4-oxazepane-2-carboxylic acid (248 mg, 1.01 mmol) and 4'-[(2S)-2-amino-2-cyanoethyl]biphenyl-4-carbonitrile (intermediate 1, 1200 mg, 0.81 mmol) were added to T3P (700 mg, 50% solution in DMF) (2 mL). TEA (640 μL, 4.54 mmol) was added, and the reaction was stirred at room temperature for 18 hours. The reaction mixture was then concentrated under reduced pressure. The resulting oil was dissolved in SiO2 and washed sequentially with 2 M aqueous hydrochloric acid, saturated aqueous sodium bicarbonate, and sodium chloride solution. The organic extract was dried (magnesium sulfate), filtered, and concentrated under reduced pressure to obtain the subtitle compound as a yellow oil, which was used in the next step without further purification.

[0271] ii) A diastereomer mixture of (2S)-N-[(1S)-1-cyano-2-(4'-cyanobiphenyl-4-yl)ethyl]-1,4-oxazepan-2-carboxamide and (2R)-N-[(1S)-1-cyano-2-(4'-cyanobiphenyl-4-yl)ethyl]-1,4-oxazepan-2-carboxamide The compound was prepared using 2-{[(1S)-1-cyano-2-(4'-cyanobiphenyl-4-yl)ethyl]carbamoyl}-1,4-oxazepane-4-carboxylate tert-butyl according to step ii) of Method A, and the marked compound was obtained as a white solid (150 mg, 50% over two steps). The isolated compound was a mixture of two diastereomers, but they did not separate. 1 ¹H NMR (400 MHz, CDCl3): δ 7.75-7.64 (m, 4H), 7.59 (dd, 2H), 7.43 (dd, 2H), 7.30-7.22 (m, 1H), 5.25-5.11 (m, 1H), 4.12-4.06 (m, 1H), 4.05-3.95 (m, 1H), 3.81-3.70 (m, 1H), 3.33 (ddd, 1H), 3.25-3.09 (m, 2H), 3.08-3.00 (m, 1H), 2.98-2.81 (m, 2H), 1.92-1.75 (m, 2H) (one exchangeable proton was not observed). LCMS (10cm_ESCI_Formic_MeCN) tR 2.58 (min) m / z 375 (MH + ).

[0272] Example 35 (2S)-N-{(1S)-1-cyano-2-[4-(4-methyl-3-oxo-1,2,3,4-tetrahydroquinoxaline-6-yl)phenyl]ethyl}-1,4-oxazepan-2-carboxamide [ka] i)(2S)-2-({(2S)-1-amino-3-[4-(4-methyl-3-oxo-1,2,3,4-tetrahydroquinoxaline-6-yl)phenyl]-1-oxopropan-2-yl}carbamoyl)-1,4-oxazepan-4-carboxylate tert-butyl 7-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-1-methylquinoxaline-2(1H)-one (boronic acid ester 2, 100 mg, 0.37 mmol) and (2S)-2-{[(2S)-1-amino-3-(4-iodophenyl)-1-oxopropan-2-yl]carbamoyl}-1,4-oxazepan-4-carboxylate tert-butyl (intermediate 4, 182 mg, 0.35 mmol) were dissolved in ACN (9 mL) and water (0.4 mL). The reaction mixture was degassed under nitrogen for 30 minutes, and then potassium carbonate (73 mg, 0.53 mmol) and Pd(dppf)Cl2·DCM (29 mg, 0.035 mmol) were added. The reaction mixture was heated at 80°C for 1 hour. The reaction was then concentrated under reduced pressure. The compound was purified by silica gel column chromatography and eluted with 8% methanol in toluene to obtain the compound in question as a brown oil (192 mg, 100%). It was used in the next step without further purification.

[0273] ii) (2S)-2-({(1S)-1-cyano-2-[4-(4-methyl-3-oxo-1,2,3,4-tetrahydroquinoxaline-6-yl)phenyl]ethyl}carbamoyl)-1,4-oxazepane-4-carboxylate tert-butyl (2S)-2-({(2S)-1-amino-3-[4-(4-methyl-3-oxo-1,2,3,4-tetrahydroquinoxaline-6-yl)phenyl]-1-oxopropan-2-yl}carbamoyl)-1,4-oxazepan-4-carboxylate tert-butyl (192 mg, 0.35 mmol) was dissolved in DCM (15 mL) and Burgess's reagent (167 mg, 0.70 mmol) was added. The reaction mixture was stirred at room temperature for 24 hours. The reaction was then transferred to a separatory funnel and washed with water. The organic extract was dried (in a phase separator cartridge) and concentrated under reduced pressure. The resulting solid was purified by silica gel column chromatography and eluted with 65% ethyl acetate in isohexane to obtain a yellow oil. Tritulate with diethyl ether to obtain the subtitle compound as an oil (101 mg, 54%). 1H NMR (400 MHz, CDCl3): δ 8.32 (s, 1H), 7.95 (d, 1H), 7.67 (d, 2H), 7.58 (dd, 1H), 7.49 (d, 1H), 7.43 (d, 2H), 7.10-7.03 (m, 1H), 5.25-5.12 (m, 1H), 4.23-4.10 (m, 3H), 3.77 (s, 3H), 3.54-3.49 (m, 3H), 3.28-3.19 (m, 3H), 2.05-1.89 (m, 2H), 1.47 (s, 9H).

[0274] iii) (2S)-N-{(1S)-1-cyano-2-[4-(4-methyl-3-oxo-1,2,3,4-tetrahydroquinoxaline-6-yl)phenyl]ethyl}-1,4-oxazepan-2-carboxamide (2S)-2-({(1S)-1-cyano-2-[4-(4-methyl-3-oxo-1,2,3,4-tetrahydroquinoxaline-6-yl)phenyl]ethyl}carbamoyl)-1,4-oxazepane-4-carboxylate tert-butyl (101 mg, 0.19 mmol) was dissolved in formic acid (2 mL) and heated at 50°C for 10 minutes on a preheated hot plate stirrer. The reaction was then concentrated under reduced pressure, dissolved in DCM, and washed with saturated sodium bicarbonate solution. The organic extract was passed through a hydrophobic frit / phase separator and concentrated under reduced pressure. The solid was purified by silica gel column chromatography and eluted with 0-2% methanolic ammonia (7N) in DCM to obtain the marked compound as a yellow solid (65 mg, 80%). 1¹H NMR (400 MHz, CDCl3): δ 8.32 (s, 1H), 7.95 (d, 1H), 7.65 (d, 2H), 7.57 (dd, 1H), 7.47 (m, 3H), 7.21 (d, 1H), 5.22 (dt, 1H), 4.11 (dd, 1H), 4.00 (dt, 1H), 3.75 (m, 5H), 3.32 (dd, 1H), 3.17 (m, 2H), 3.06 (dd, 1H), 2.99-2.87 (m, 2H), 1.89-1.81 (m, 2H) (two exchangeable protons were not observed). LCMS (10cm_ESCI_Formic_MeCN) tR 2.38 (min) m / z 432 (MH + ).

[0275] Example 36 (2S)-2-[(3S,4E)-6-(2,3-dihydro-1H-indole-1-yl)-6-oxohexa-4-en-3-yl]-1,4-oxazepan-2-carboxamide / trifluoroacetic acid [ka] i) (2S)-2-{[(3S,4E)-6-(2,3-dihydro-1H-indole-1-yl)-6-oxohexa-4-en-3-yl]carbamoyl}-1,4-oxazepane-4-carboxylate tert-butyl [ka] At room temperature, (2S)-4-(tert-butoxycarbonyl)-1,4-oxazepan-2-carboxylic acid (intermediate 3, 1.25 g, 5.10 mmol), [(1S,2E)-4-(2,3-dihydro-1H-indole-1-yl)-1-ethyl-4-oxo-buten-1-yl]amine trifluoroacetate (intermediate 6 from International Publication No. 2012 / 109415, 1.76 g, 5.10 mmol), and DiPEA (4.45 ml, 25.5 mmol) were mixed with HATU (2.33 g, 6.12 mmol). The resulting mixture was stirred at room temperature for 4 hours. The reaction mixture was diluted with DCM (100 mL) and washed sequentially with 0.1 M aqueous HCl (100 mL), saturated aqueous NaHCO3 (100 mL), and saturated brine (100 mL). The organic layer was dried over Na2SO4, filtered, and evaporated to obtain the secondary product (1.50 g, 64%). LC-MS m / z 358 (M-Boc+H + The crude product sample (190 mg, 0.42 mmol) was purified by preparative chiral HPLC using a CHIRALPAK IC-3 column, and homogeneous elution was performed using 50% EtOH in hexane as the eluent. The fraction containing the desired compound was evaporated to dryness, and the secondary product was obtained as a colorless oil (180 mg, 95%). LC-MS m / z 358 (M-Boc+H + ). At room temperature, (2S)-2-{[(3S,4E)-6-(2,3-dihydro-1H-indole-1-yl)-6-oxohexa-4-en-3-yl]carbamoyl}-1,4-oxazepane-4-carboxylate tert-butyl (180 mg, 0.39 mmol) was added to DCM (10 mL) with TFA (2 mL, 26.0 mmol). The resulting solution was stirred at room temperature for 4 hours. The solvent was removed under reduced pressure. The crude product was purified by preparative flash (C18 column) using a decreasing polar mixture of water (containing 0.1% TFA) and MeCN as the eluent. The fraction containing the desired product was dried by lyophilization to obtain the labeled product as a white solid (100 mg, 54%). 1H NMR (300 MHz, DMSO-d6): δ 8.80-9.10 (m, 2H), 8.30 (d,1H), 8.15 (d, 1H), 7.10-7.30 (m, 2H), 6.95-7.10 (m, 1H), 6.70-6.85 (m, 1H), 6.45 (d, 1H), 4.10-4.70 (m, 4H), 3.90-4.10 (m, 1H), 3.75-3.85 (m, 1H), 3.55-3.70 (m, 1H), 3.05-3.40 (m, 5H), 1.90-2.10 (m, 2H), 1.50-1.75 (m, 2H), 0.85 (t, 3H). LCMS m / z 358 (MH) + ).

[0276] Example 37 (2S)-2-[(2E,4S)-1-(2,3-dihydro-1H-indole-1-yl)-6-methyl-1-oxohepta-2-en-4-yl]-1,4-oxazepan-2-carboxamide trifluoroacetate [ka] i) (2S)-2-{[(2E,4S)-1-(2,3-dihydro-1H-indole-1-yl)-6-methyl-1-oxohepta-2-en-4-yl]carbamoyl}-1,4-oxazepane-4-carboxylate tert-butyl [ka] (2S)-4-(tert-butoxycarbonyl)-1,4-oxazepan-2-carboxylic acid (intermediate 3, 150 mg, 0.61 mmol), [(1S,2E)-4-(2,3-dihydro-1H-indol-1-yl)-1-(2-methylpropyl)-4-oxo-2-buten-1-yl]amine trifluoroacetate (intermediate 13 from International Publication No. 2012 / 109415, 174 mg, 0.47 mmol), and DiPEA (0.427 mL, 2.45 mmol) were mixed with HATU (465 mg, 1.22 mmol) at 0°C. The resulting solution was stirred at room temperature for 2.5 hours. The reaction mixture was evaporated to dryness, redissolved in siRNA (25 mL), and washed sequentially with saturated aqueous solution of NH4Cl (20 mL x 4), saturated brine (20 mL x 3), and water (20 mL x 3). The organic layer was dried over Na2SO4, filtered, and evaporated to obtain the crude secondary product as a yellow oil (200 mg, 67%). LC-MS m / z 486 (MH) + The crude product was used in the next step without further purification. (2S)-2-{[(2E,4S)-1-(2,3-dihydro-1H-indol-1-yl)-6-methyl-1-oxohepta-2-en-4-yl]carbamoyl}-1,4-oxazepane-4-carboxylate tert-butyl (200 mg, 0.41 mmol) was added to DCM (5.0 mL) at 0°C with TFA (0.635 mL, 8.24 mmol). The resulting solution was stirred at room temperature for 2 hours. The solvent was removed under reduced pressure. The crude product was purified by preparative HPLC (Waters XBridge Prep C18 OBD column, 5 μS silica, 19 mm diameter, 150 mm length) using a decreasing polar mixture of water (containing 0.5% TFA) and MeCN as the eluent. The fraction containing the desired product was evaporated to dryness to obtain the labeled product as a yellow gum (130 mg, 63%). LC-MS m / z 386 (MH + ). 1¹H-NMR (300 MHz, CD3OD): δ 8.15 (1H, d), 7.10-7.30 (2H, m), 7.05 (1H, t), 6.75-6.90 (1H, m), 6.50 (1H, d), 4.60-4.80 (1H, m), 4.40-4.55 (1H, m), 4.10-4.30 (3H, m), 3.70-3.95 (2H, m), 3.15-3.50 (5H, m), 2.05-2.25 (2H, m), 1.40-1.70 (3H, m), 0.95 (6H, t), 1.35 (1H, d) (two exchangeable protons were not observed).

[0277] Pharmacological activity Test A1: Fluorescence assay of recombinant human (RH) DPP1 The activity of DPP1 was determined by measuring the enzymatic release of aminomethylcoumarin (AMC) from the peptide substrate (H-Gly-Arg-AMC), which resulted in increased fluorescence intensity at λex=350nm and λem=450nm. The assay was performed in a black 384-well plate with a final volume of 50 μl at 22°C. The assay conditions included: 25 mM piperazine buffer pH 5.0; 50 mM NaCl, 5 mM DTT; 0.01% (v / v) Triton X-100; 100 μM H-Gly-Arg-AMC and rhDPP1 (approximately 50 pM). Potential inhibitors were prepared in DMSO and diluted in the assay so that the final concentration did not exceed 1% (v / v) DMSO. Ten consecutive half-log dilutions of the inhibitor (typically with a maximum concentration of 10 μM) were tested, and the pIC was calculated using the four-parameter logistic equation with a nonlinear curve fitting method. 50The following was calculated. The standard DPP1 inhibitor, 4-amino-N-[(1S)-1-cyano-2-(4'-cyanobiphenyl-4-yl)ethyl]tetrahydro-2H-pyran-4-carboxamide (Example 3 of International Publication No. 2010 / 128324), was used as a positive control in the assay. As specified, the inhibitor was pre-incubated with rhDPP1 for 30-60 minutes, then the peptide substrate was added and the reaction was initiated at 22°C for a further 60 minutes. Immediately thereafter, the plates were read using a fluorescence plate reader with the emission and excitation wavelengths described above [modified from Kam, CM, Gotz, MG, Koot, G, McGuire, MJ, Thiele, DL, Hudig, D & Powers, JC (2004). Arch Biochem Biophys, 427, 123-134 & McGuire, MJ, Lipsky, PE & Thiele, DL (1992). Arch Biochem Biophys, 295, 280-288]. The results obtained are shown in Table 11 below (Examples 1-35).

[0278] Study A2: Fluorescence assay of recombinant human (RH) DPP1 The activity of DPP1 was determined by measuring the enzymatic release of aminomethylcoumarin (AMC) from the peptide substrate (H-Gly-Arg-AMC), which resulted in increased fluorescence intensity at λex=350nm and λem=450nm. The assay was performed in a black 384-well plate at room temperature with a final volume of 10 μl. The assay conditions included: 25 mM piperazine buffer pH 5.0; 50 mM NaCl, 5 mM DTT; 0.005% (v / v) Triton X-100; 50 μM H-Gly-Arg-AMC and 96.4 pM rhDPP1. Potential inhibitors were diluted with DMSO to obtain 100-fold dilutions of the final assay concentration. The compounds were tested at 10 different concentrations using a half-log dilution step (typically with a maximum concentration of 1 μM) with a final DMSO concentration of 1% (v / v). Following the prescribed procedure, the inhibitor was pre-incubated with rhDPP1 for 30 minutes, then the peptide substrate was added and the reaction was initiated for another 30 minutes. After incubation, the plate was read using a fluorescence plate reader with the above emission and excitation wavelengths. The pIC was calculated using the 4-parameter logistic equation with a nonlinear curve fitting method. 50 The appropriate inhibitor was determined (Smartfit, Geneda Screener®). The standard DPP1 inhibitor, 4-amino-N-[(1S)-1-cyano-2-(4'-cyanobiphenyl-4-yl)ethyl]tetrahydro-2H-pyran-4-carboxamide (Example 3 of International Publication No. 2010 / 128324) was used as a positive control. [Adapted from Kam, CM, Gotz, MG, Koot, G, McGuire, MJ, Thiele, DL, Hudig, D & Powers, JC (2004). Arch Biochem Biophys, 427, 123-134 & McGuire, MJ, Lipsky, PE & Thiele, DL (1992). Arch Biochem Biophys, 295, 280-288]. The results obtained are shown in Table 11 below (Examples 36-37).

[0279] [Table 13-1] [Table 13-2]

[0280] Aortic connection Numerous compounds have been documented in the literature as being selectively maintained within the aorta in quantitative whole-body autoradiography (QWBA) studies, which induce contingent hyperstructural changes when examined by electron microscopy (see, e.g., muzolimin (Schmidt et al. 1984, Biochem. Pharmacol., 33, 1915-1921)). Furthermore, α-aminoamidonitrile 4-amino-N-[(1S)-1-cyano-2-(4'-cyanobiphenyl-4-yl)ethyl]tetrahydro-2H-pyran-4-carboxamide (Example 3 in International Publication 2010 / 128324), described as a DPP1 inhibitor, has been shown to exhibit high aortic retention levels in rat QWBA studies. To aid in the design of DPP1 inhibitors that reduce the risk of binding to elastin-rich tissues (e.g., the aorta), the following in vitro competitive aortic binding assay (Sample B) was developed to facilitate the selection process. The reference compound and the selected compounds described herein were tested using Method B, and the results are shown in Table 12.

[0281] Study B: In vitro competitive aortic coupling assay Aortic homogenates were prepared from the thoracic aorta of Han Wistar rats. Freshly isolated thoracic aortas were frozen, then thawed, and the inelastic material was stripped. The stripped aorta was then weighed, cut into small pieces, and first homogenized using a rotor-stator homogenizer; then homogenized in Puck's saline (137 mM NaCl, 5.37 mM KCl, 4.17 mM NaHCO3, and 5.55 mM D-glucose) using a Doughs homogenizer with a loose fit followed by a tight fit. The homogenate concentration was adjusted to 30 mg / mL with Puck's saline, and aliquots were stored at -80°C until use. Positive and negative control compounds, as well as the test compound, were prepared to 100 mM with DMSO and added to 1 mL aliquots of aortic homogenate in Puck's saline to a final concentration of 100 μM. The homogenate samples were pre-incubated overnight at 37°C with the test compound while rotating. Next, all samples were treated with [ 14[C]4-amino-N-[(1S)-1-cyano-2-(4'-cyanobiphenyl-4-yl)ethyl]tetrahydro-2H-pyran-4-carboxamide was added to a final concentration of 100 μM, and the samples were incubated at 37°C for a further 2 hours with rotation. 10 mL of acetone was added to precipitate proteins from each sample, and the mixture was pre-cooled to -20°C. The samples were left at -20°C overnight to complete precipitation. The precipitate was pelletized by centrifugation at 4,500 × g for 20 minutes at 4°C, and aliquots of the supernatant were taken for analysis, with the remaining supernatant discarded. The precipitate was washed by resuspending it in 10 mL of 80% methanol in distilled water, and repelled by centrifugation at 4,500 × g for 20 minutes at 4°C. The washing was repeated four times with 80% methanol and two more times with 100% methanol, with aliquots of the supernatant taken for analysis each time. After the final wash, the precipitate was air-dried and dissolved overnight in 1 mL of NCSII Tissue Solubiliser. A 1 mL aliquot of the supernatant was added to 5 mL of Ultima Gold scintillation solution (Perkin Elmer, MA, USA), and 1 mL of the solubilized pellet was added to 5 mL of Hionic-Fluor scintillation solution (Perkin Elmer, MA, USA). The radioactivity of the sample was measured using a Beckman LS6500 multi-purpose scintillation counter (Beckman Coulter, IN, USA). Each time, 4-amino-N-[(1S)-1-cyano-2-(4'-cyanobiphenyl-4-yl)ethyl]tetrahydro-2H-pyran-4-carboxamide was used as a positive control, and N-(1-{(3R)-3-(3,5-difluorophenyl)-3-[1-(methylsulfonyl)piperidine-4-yl]propyl}piperidine-4-yl)-N-ethyl-2-[4-(methylsulfonyl)phenyl]acetamide (compound 1, International Publication No. 2006 / 001751) and DMSO vehicle were used as negative controls. Two samples were tested for each compound in each experiment, and at least two experiments were performed for each test compound.The average radioactivity of each sample pre-incubated with the DMSO vehicle control was set as 100% bound, and the results of samples pre-incubated with other compounds were expressed as the difference from the vehicle control in percentage. The significance of the difference from the vehicle control was calculated using one-way ANOVA and Bonferroni multiple comparison tests.

[0282] The results obtained are shown in Table 12 below. The results were quantified and divided into four categories: strong binder, moderate binder, binder, and no binder.

[0283] [Table 14-1] [Table 14-2]

[0284] [ 14 C]4-amino-N-[(1S)-1-cyano-2-(4'-cyanobiphenyl-4-yl)ethyl]tetrahydro-2H-pyran-4-carboxamide [ka] i) 4-bromobenzo-[14C]-nitrile 1-Methylpyrrolidine-2-one (4 mL) with 1-bromo-4-iodobenzene (473 mg, 1.67 mmol) and [ 14 Copper(I) cyanide (1850 MBq, 77 mg, 0.84 mmol) was dissolved and heated at 150°C for 3 hours using ultrasound. The reaction was diluted with SiO2 (150 ml) and washed with 2% ferric chloride aqueous solution (100 ml), 2% w / v sodium thiosulfate aqueous solution (100 ml), and saturated brine (25 mL x 3). The organic material was passed through a phase separator to remove the solvent and obtain the crude product. The crude product was purified by silica gel column chromatography and eluted with 2% SiO2 in isoheptane to obtain the marked compound as a white solid (442 MBq, 37 mg, 24%).

[0285] ii)(S)-4-(1-amino-3-(4'-[14C]-cyanobiphenyl-4-yl)-1-oxopropane-2-ylcarbamoyl)tetrahydro-2H-pyran-4-ylcarbamate tert-butyl (S)-4-(1-amino-1-oxo-3-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)propan-2-ylcarbamoyl)tetrahydro-2H-pyran-4-ylcarbamate tert-butyl (243 mg, 0.47 mmol), Pd-118 (30.6 mg, 0.05 mmol) and potassium carbonate (195 mg, 1.41 mmol) were added to a flask under a nitrogen atmosphere. 4-bromobenzo-[ in degassed ACN (6 mL) was added to the reaction flask. 14 [C]-nitrile (973 MBq, 86 mg, 0.47 mmol) was added, followed by the addition of water (3 mL). The mixture was heated under nitrogen at 73 °C for 4 hours and left overnight at room temperature. The reaction was diluted with water (50 mL), and the product was extracted in DCM (25 mL x 4). The combined organic matter was washed with saturated brine (50 mL), and the organic matter portion was passed through a phase separator containing magnesium sulfate. The organic matter was concentrated under vacuum to obtain a dark brown oil. The crude substance was purified by silica gel column chromatography and eluted with 0-100% siRNA in heptane to obtain a gum, which was tritulated with ether / heptane to obtain the marked compound as an off-white solid (802 MBq, 189 mg, 82%). m / z (ES+) 395 [M+2H-BOC] + .

[0286] iii)(S)-4-(1-cyano-2-(4'-[14C]-cyanobiphenyl-4-yl)ethylcarbamoyl)tetrahydro-2H-pyran-4-ylcarbamate tert-butyl (S)-4-(1-amino-3-(4'-[14C]-cyanobiphenyl-4-yl)-1-oxopropane-2-ylcarbamoyl)tetrahydro-2H-pyran-4-ylcarbamate tert-butyl (802 MBq, 189 mg, 0.38 mmol) was dissolved in DCM (4 mL) and stirred at room temperature under nitrogen. Burgess reagent (137 mg, 0.57 mmol) was added and the reaction was stirred for 6.5 hours. The crude mixture was purified by silica gel column chromatography and eluted with 25-100% siRNA in heptane to obtain the marked compound as a white solid (714 MBq, 164 mg, 90%). 1 H NMR (500 MHz, DMSO-d6): δ 1.38 (s, 9H), 1.55 - 1.77 (m, 2H), 1.84 - 2.02 (m, 1H), 3.07 - 3.25 (m, 3H), 3.43 - 3.53 (m, 1H), 3.54 - 3.62 (m, 1H), 5.04 - 5.13 (m, 1H), 7.04 (s, 1H), 7.43 (d, 2H), 7.71 (d, 2H), 7.87 (d, 2H), 7.93 (d, 2H), 8.46 (s, 1H). m / z (ES-) 475 [MH] - .

[0287] iv)(S)-4-amino-N-(1-cyano-2-(4'-[14C]-cyanobiphenyl-4-yl)ethyl)tetrahydro-2H-pyran-4-carboxamide To a preheated formic acid solution (500 μl, 13.04 mmol, 50°C), (S)-4-(1-cyano-2-(4'-[14C]-cyanobiphenyl-4-yl)ethylcarbamoyl)tetrahydro-2H-pyran-4-ylcarbamate tert-butyl (133 MBq, 29 mg, 0.06 mmol) was added, and the reaction was heated at 50°C for 15 minutes with stirring. The reaction was rapidly cooled and added to a cooled mixture of saturated sodium bicarbonate (5 ml) and DCM (5 ml). The aqueous portion was further washed with two aliquots of DCM (5 ml), the combined organic material was washed with water (10 ml), and dried over sodium sulfate. The organic material was removed to obtain a colorless oil, which was triturated with ether to obtain a white solid. The crude mixture was purified by silica gel column chromatography and eluted with 0-2% methanol in DCM to obtain the labeled compound (93 MBq, 68%), which was stored as a MeCN solution. 1 H NMR (500 MHz, DMSO-d6): δ 1.12 (d, 1H), 1.20 (d, 1H), 1.73 (ddd, 1H), 1.89 (ddd, 1H), 3.18 - 3.25 (m, 2H), 3.45 (dt, 1H), 3.53 - 3.66 (m, 3H), 5.02 (t, 1H), 7.43 (d, 2H), 7.71 (d, 2H), 7.89 (dd, 4H). m / z (ES+) 377 [M+H] + .

[0288] This application also includes the following aspects. [Aspect 1] Equation (I): [ka] [In the formula, R 1 teeth, [ka] and; R 2 These are hydrogen, F, Cl, Br, and OSO2C. 1-3 Alkyl or C1-3 Selected from alkyl groups; R 3 These are hydrogen, F, Cl, Br, CN, CF3, SO2C 1-3 Alkyl, CONH2, or SO2NR 4 R 5 Selected from (where R 4 and R 5 (These together with the nitrogen atom to which they are bonded form an azetidine ring, a pyrrolidine ring, or a piperidine ring); or R 1 teeth, [ka] Selected from; X is selected from O, S, or CF2; Y is selected from O or S; Q is selected from CH or N; R 6 C 1-3 Selected from alkyl (where C 1-3 The alkyl group may be substituted with 1, 2, or 3 F atoms, and also OH,OC 1-3 Alkyl, N(C 1-3 It may be substituted with one substituent selected from alkyl)2, cyclopropyl, or tetrahydropyran); R 7 [Selected from hydrogen, F, Cl, or CH3] The compound indicated by or a pharmaceutically acceptable salt thereof. [Aspect 2] R 1 but [ka] That is, The compound described in Embodiment 1, or a pharmaceutically acceptable salt thereof. [Aspect 3] X is O; R 6 C 1-3 It is alkyl; R 7 That is hydrogen. The compound according to embodiment 1 or 2, or a pharmaceutically acceptable salt thereof. [Aspect 4] (2S)-N-[(1S)-1-cyano-2-(4'-cyanobiphenyl-4-yl)ethyl]-1,4-oxazepan-2-carboxamide; (2S)-N-{(1S)-1-cyano-2-[4-(3-methyl-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl)phenyl]ethyl}-1,4-oxazepan-2-carboxamide; (2S)-N-{(1S)-1-cyano-2-[4-(3,7-dimethyl-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl)phenyl]ethyl}-1,4-oxazepan-2-carboxamide; 4'-[(2S)-2-cyano-2-{[(2S)-1,4-oxazepan-2-ylcarbonyl]amino}ethyl]biphenyl-3-ylmethanesulfonate; (2S)-N-{(1S)-1-cyano-2-[4-(3-methyl-1,2-benzoxazol-5-yl)phenyl]ethyl}-1,4-oxazepan-2-carboxamide; (2S)-N-{(1S)-1-cyano-2-[4'-(trifluoromethyl)biphenyl-4-yl]ethyl}-1,4-oxazepan-2-carboxamide; (2S)-N-[(1S)-1-cyano-2-(3',4'-difluorobiphenyl-4-yl)ethyl]-1,4-oxazepan-2-carboxamide; (2S)-N-{(1S)-1-cyano-2-[4-(6-cyanopyridine-3-yl)phenyl]ethyl}-1,4-oxazepan-2-carboxamide; (2S)-N-{(1S)-1-cyano-2-[4-(4-methyl-3-oxo-3,4-dihydro-2H-1,4-benzothiadin-6-yl)phenyl]ethyl}-1,4-oxazepan-2-carboxamide; (2S)-N-{(1S)-1-cyano-2-[4-(3-ethyl-7-methyl-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl)phenyl]ethyl}-1,4-oxazepan-2-carboxamide; (2S)-N-[(1S)-1-cyano-2-{4-[3-(2-hydroxy-2-methylpropyl)-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl]phenyl}ethyl]-1,4-oxazepan-2-carboxamide; (2S)-N-[(1S)-1-cyano-2-{4-[3-(2,2-difluoroethyl)-7-fluoro-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl]phenyl}ethyl]-1,4-oxazepan-2-carboxamide; (2S)-N-[(1S)-1-cyano-2-(4-{3-[2-(dimethylamino)ethyl]-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl}phenyl)ethyl]-1,4-oxazepan-2-carboxamide; (2S)-N-{(1S)-1-cyano-2-[4-(3,3-difluoro-1-methyl-2-oxo-2,3-dihydro-1H-indole-6-yl)phenyl]ethyl}-1,4-oxazepan-2-carboxamide; (2S)-N-{(1S)-1-cyano-2-[4-(7-fluoro-3-methyl-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl)phenyl]ethyl}-1,4-oxazepan-2-carboxamide; (2S)-N-{(1S)-1-cyano-2-[4-(3-ethyl-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl)phenyl]ethyl}-1,4-oxazepan-2-carboxamide; (2S)-N-[(1S)-1-cyano-2-{4-[3-(cyclopropylmethyl)-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl]phenyl}ethyl]-1,4-oxazepan-2-carboxamide; (2S)-N-[(1S)-1-cyano-2-{4-[3-(2-methoxyethyl)-2-oxo-2,3-dihydro-1,3-benzothiazole-5-yl]phenyl}ethyl]-1,4-oxazepan-2-carboxamide; (2S)-N-[(1S)-1-cyano-2-{4-[2-oxo-3-(propan-2-yl)-2,3-dihydro-1,3-benzoxazole-5-yl]phenyl}ethyl]-1,4-oxazepan-2-carboxamide; (2S)-N-{(1S)-1-cyano-2-[4-(4-methyl-3-oxo-3,4-dihydro-2H-1,4-benzoxazine-6-yl)phenyl]ethyl}-1,4-oxazepan-2-carboxamide; (2S)-N-[(1S)-1-cyano-2-{4-[3-(2-methoxyethyl)-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl]phenyl}ethyl]-1,4-oxazepan-2-carboxamide; (2S)-N-{(1S)-1-cyano-2-[4-(5-cyanothiophen-2-yl)phenyl]ethyl}-1,4-oxazepan-2-carboxamide; (2S)-N-[(1S)-2-(4'-carbamoyl-3'-fluorobiphenyl-4-yl)-1-cyanoethyl]-1,4-oxazepan-2-carboxamide; (2S)-N-{(1S)-1-cyano-2-[4-(1-methyl-2-oxo-1,2-dihydroquinoline-7-yl)phenyl]ethyl}-1,4-oxazepan-2-carboxamide; (2S)-N-[(1S)-1-cyano-2-{4-[2-oxo-3-(tetrahydro-2H-pyran-4-ylmethyl)-2,3-dihydro-1,3-benzoxazole-5-yl]phenyl}ethyl]-1,4-oxazepan-2-carboxamide; (2S)-N-{(1S)-2-[4-(7-chloro-3-methyl-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl)phenyl]-1-cyanoethyl}-1,4-oxazepan-2-carboxamide; (2S)-N-[(1S)-1-cyano-2-{4-[3-(2,2-difluoroethyl)-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl]phenyl}ethyl]-1,4-oxazepan-2-carboxamide; (2S)-N-[(1S)-1-cyano-2-{4-[2-oxo-3-(2,2,2-trifluoroethyl)-2,3-dihydro-1,3-benzoxazole-5-yl]phenyl}ethyl]-1,4-oxazepan-2-carboxamide; (2S)-N-{(1S)-1-cyano-2-[4-(3-methyl-2-oxo-2,3-dihydro-1,3-benzothiazole-5-yl)phenyl]ethyl}-1,4-oxazepan-2-carboxamide; (2S)-N-{(1S)-1-cyano-2-[4'-(methylsulfonyl)biphenyl-4-yl]ethyl}-1,4-oxazepan-2-carboxamide; (2S)-N-{(1S)-2-[4'-(azetidine-1-ylsulfonyl)biphenyl-4-yl]-1-cyanoethyl}-1,4-oxazepan-2-carboxamide; (2S)-N-[(1S)-1-cyano-2-(4'-fluorobiphenyl-4-yl)ethyl]-1,4-oxazepan-2-carboxamide; (2S)-N-{(1S)-2-[4-(1,3-benzothiazole-5-yl)phenyl]-1-cyanoethyl}-1,4-oxazepan-2-carboxamide; (2S)-N-[(1S)-1-cyano-2-(4'-cyanobiphenyl-4-yl)ethyl]-1,4-oxazepan-2-carboxamide; or (2S)-N-{(1S)-1-cyano-2-[4-(4-methyl-3-oxo-1,2,3,4-tetrahydroquinoxaline-6-yl)phenyl]ethyl}-1,4-oxazepan-2-carboxamide A compound represented by formula (I) according to embodiment 1, selected from the above, or a pharmaceutically acceptable salt thereof. [Aspect 5] [ka] The compound according to Embodiment 1, which is (2S)-N-{(1S)-1-cyano-2-[4-(3-methyl-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl)phenyl]ethyl}-1,4-oxazepan-2-carboxamide, or a pharmaceutically acceptable salt thereof. [Aspect 6] [ka] The compound according to embodiment 1, which is (2S)-N-{(1S)-1-cyano-2-[4-(3-methyl-2-oxo-2,3-dihydro-1,3-benzoxazole-5-yl)phenyl]ethyl}1,4-oxazepan-2-carboxamide. [Aspect 7] A pharmaceutical composition comprising a compound represented by formula (I) as described in any one of embodiments 1 to 6, and a pharmaceutically acceptable adjuvant, diluent, or carrier. [Aspect 8] A compound represented by formula (I) as described in any one of embodiments 1 to 6 for use in treatment. [Aspect 9] A compound represented by formula (I) as described in any one of embodiments 1 to 6 for use in the treatment of asthma or chronic obstructive pulmonary disease. [Aspect 10] Use of a compound represented by formula (I) as described in any one of embodiments 1 to 6 in the manufacture of a therapeutic agent for asthma or chronic obstructive pulmonary disease. [Aspect 11] A method for treating asthma or chronic obstructive pulmonary disease in a patient, comprising administering a therapeutically effective amount of a compound represented by formula (I) as described in any one of embodiments 1 to 6. [Aspect 12] A compound represented by formula (I) as described in any one of embodiments 1 to 6, • Nonsteroidal glucocorticoid receptor agonists; • Selective β2 adrenoceptor agonists; • Phosphodiesterase inhibitors; • Protease inhibitors; • Glucocorticoids; • Anticholinergics; • Modulators of chemokine receptor function; and • Inhibitors of kinase function A combination drug with one or more drugs selected independently from the original drug.

Claims

[Claim 1] The invention described in the specification or drawings.

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