Aromatic acetylene derivative as well as preparation method and application thereof

Abstract

The invention relates to an aromatic acetylene derivative, a preparation method thereof and application of a pharmaceutical composition containing the derivative in medicine. Specifically, the present invention relates to an aromatic acetylene derivative represented by general formula (I), a preparation method and a pharmaceutically acceptable salt thereof, and uses thereof as a therapeutic agent, especially an LPXC inhibitor, the definition of each substituent in the general formula (I) being the same as the definition in the specification, # imgabs0 #

Description

Aromatic acetylene derivatives and their preparation method and use Technical Field

[0001] The present invention relates to an aromatic acetylene derivative, a preparation method thereof, a pharmaceutical composition containing the derivative, and use of the derivative as a therapeutic agent, in particular as an LPXC inhibitor. Background Art

[0002] The 1930s to 1960s marked the golden age of antibiotic development. Since then, antibiotics have been widely used worldwide, but bacterial resistance has also emerged, and drug-resistant bacteria have become a major threat to human health. Multidrug-resistant Gram-negative bacteria are one of the main pathogens of infection. Currently, there is a severe shortage of clinical drugs to treat multidrug-resistant Gram-negative infections, and highly toxic drugs are still used. Although bacterial resistance has been a hot topic in the international medical community in recent years, research and development has been slow, and few compounds have entered clinical research at home and abroad. Therefore, finding a new antibacterial drug for Gram-negative bacteria is an important issue that needs to be addressed urgently.

[0003] UDP-3-O-(R-3-hydroxymyristoyl)-N-acetylglucosamine deacetylase (LPXC) is a Zn-dependent + LPXC is a metalloenzyme that is the first, rate-limiting enzyme in the synthesis of lipid A. Lipid A is a key component of the outer membrane of Gram-negative bacteria. It anchors lipopolysaccharide to the outer membrane, maintaining the integrity of the cell itself. It also acts as a hydrophobic barrier, preventing the entry of external factors such as antibiotics into the cell, protecting the bacteria from harm. Furthermore, lipid A is also the active component of bacterial endotoxins, which enter the bloodstream through the intestinal mucosa, activating the body's immune response and even causing severe septic shock. This is also the reason why Gram-negative bacteria are pathogenic. Therefore, by inhibiting LPXC, the biosynthesis of lipid A in Gram-negative bacteria can be inhibited, effectively controlling Gram-negative infections.

[0004] To date, much of the knowledge regarding the structure and properties of LPXC has been gained through the isolation, purification, and characterization of LPXC crystals from Escherichia coli, Pseudomonas aeruginosa, and thermophilic bacteria. These three different sources of LPXC exhibit highly similar structures, each containing two domains with the active region located at their junction. Each domain comprises an α-helix and a β-sheet, with the β-sheet enclosing the α-helix to form a "β-α-α-β" sandwich structure. Although the amino acid sequences of these two domains differ slightly, they share a common spatial structure. Furthermore, each domain has a corresponding insertion region composed of β-sheets, forming distinct functional domains. Studies have shown that LPXC shares a high degree of homology among Gram-negative bacteria and shares no common sequences with various mammalian enzymes. From a biological perspective, due to its unique advantages of broad spectrum and low toxicity, inhibiting LPXC would be an ideal target for antibacterial drug development.

[0005] Currently, no new LPXC inhibitors have been marketed. The small molecule compound RC-01, developed by Toyama Chemical Co., Ltd., has entered Phase I clinical trials. Other pharmaceutical companies, such as Novartis and Taiwan's Taisho Pharmaceutical, are still in the preclinical stage. While the research and application of LPXC inhibitors have made some progress, significant room for improvement remains, and continued research and development of new LPXC inhibitors is necessary.

[0006] Summary of the Invention

[0007] In order to solve the above technical problems, the present invention provides an aromatic acetylene derivative represented by general formula (I) or its stereoisomers, tautomers or pharmaceutically acceptable salts:

[0008] in:

[0009] W is selected from O, S(O) r or N;

[0010] Ring C is selected from a 5- to 10-membered spiro heterocyclic group or a monocyclic heterocyclic group, wherein the monocyclic heterocyclic group contains a double bond;

[0011] X, Y, Z, Q are each independently selected from CR A or N atoms, and at most two atoms among X, Y, Z, and Q are N atoms at the same time;

[0012] R A is selected from hydrogen, halogen, hydroxy, cyano, alkyl or alkoxy; wherein the alkyl or alkoxy is optionally further substituted by one or more substituents selected from halogen, hydroxy, cyano, alkyl or alkoxy;

[0013] R 1is selected from the group consisting of: absence, hydrogen, hydroxy, cyano, halogen, alkyl, alkoxy, cycloalkyl, heterocyclic, aryl, heteroaryl, -C(O)R 3 、-C(O)NR 6 R 7 or -S(O) r R 4 wherein the alkoxy, alkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl is optionally further substituted by one or more R B replaced by;

[0014] R 3 、R 4 Each is independently selected from hydrogen atom, hydroxyl, alkyl, cycloalkyl, aryl, heteroaryl or heterocyclic group; wherein the alkyl, cycloalkyl, aryl, heteroaryl or heterocyclic group is optionally further substituted by one or more selected from hydroxyl, halogen, nitro, cyano, alkyl, alkoxy, haloalkyl, haloalkoxy, cycloalkyl, heterocyclic group, aryl, heteroaryl, =O, -C(O)R 8 、-C(O)OR 8 、-OC(O)R 8 、-NR 9 R 10 、-C(O)NR 9 R 10 、-SO2NR 9 R 10 or -NR 9 C(O)R 10 substituted by a substituent;

[0015] R B are each independently selected from cyano, halogen, alkyl, hydroxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, -OR 5 、-C(O)R 5 、-C(O)OR 5 、-NHC(O)R 5 、-NHC(O)OR 5 、-NR 6 R 7 、-C(O)NR 6 R 7 、-CH2NHC(O)OR 5 、-CH2NR 6 R 7 or -S(O) r R 5 wherein the alkyl, cycloalkyl, heterocyclic, aryl or heteroaryl is optionally further substituted by one or more selected from hydroxy, halogen, nitro, cyano, alkyl, alkoxy, haloalkyl, haloalkoxy, cycloalkyl, heterocyclic, aryl, heteroaryl, =O, -C(O)R 8、-C(O)OR 8 、-OC(O)R 8 、-NR 9 R 10 、-C(O)NR 9 R 10 、-SO2NR 9 R 10 or -NR 9 C(O)R 10 substituted by a substituent;

[0016] Or, two R B Form a -C(=O)- with the same carbon atom to which it is attached;

[0017] R 2 are the same or different, each independently selected from hydroxy, cyano, halogen, alkyl or alkoxy; wherein the alkyl or alkoxy is optionally further substituted with one or more substituents selected from halogen, hydroxy, cyano, alkyl or alkoxy;

[0018] R 5 Each is independently selected from a hydrogen atom, an alkyl group, a cycloalkyl group, a heterocyclic group, an aryl group or a heteroaryl group, wherein the alkyl group, the cycloalkyl group, the heterocyclic group, the aryl group or the heteroaryl group is optionally further substituted by one or more hydroxyl groups, halogen groups, nitro groups, cyano groups, alkyl groups, alkoxy groups, haloalkyl groups, haloalkoxy groups, cycloalkyl groups, heterocyclic groups, aryl groups, heteroaryl groups, =O, -C(O)R 8 、 -C(O)OR 8 、-OC(O)R 8 、-NR 9 R 10 、-C(O)NR 9 R 10 、-SO2NR 9 R 10 or -NR 9 C(O)R 10 substituted by a substituent;

[0019] R 6 and R 7 Each is independently selected from hydrogen, hydroxy, halogen, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl or heteroaryl, wherein the alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl or heteroaryl is optionally further substituted by one or more selected from hydroxy, halogen, nitro, cyano, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, =O, -C(O)R 8 、-C(O)OR 8 、-OC(O)R 8 、-NR 9 R 10 、-C(O)NR9 R 10 、-SO2NR 9 R 10 or -NR 9 C(O)R 10 substituted by a substituent;

[0020] Or, R 6 and R 7 Together with the atoms to which they are attached, they form a 4-8 membered heterocyclic group, wherein the 4-8 membered heterocyclic group contains one or more N, O or S(O)r, and the 4-8 membered heterocyclic group is optionally further substituted by one or more selected from hydroxy, halogen, nitro, cyano, alkyl, alkoxy, cycloalkyl, heterocyclic group, aryl, heteroaryl, =O, -C(O)R 8 、-C(O)OR 8 、-OC(O)R 8 、-NR 9 R 10 、-C(O)NR 9 R 10 、-SO2NR 9 R 10 or -NR 9 C(O)R 10 substituted by a substituent;

[0021] R 8 、R 9 and R 10 Each is independently selected from a hydrogen atom, an alkyl group, an amino group, a cycloalkyl group, a heterocyclic group, an aryl group or a heteroaryl group, wherein the alkyl group, the cycloalkyl group, the heterocyclic group, the aryl group or the heteroaryl group is optionally further substituted with one or more substituents selected from a hydroxyl group, a halogen group, a nitro group, an amino group, a cyano group, an alkyl group, an alkoxy group, a cycloalkyl group, a heterocyclic group, an aryl group, a heteroaryl group, a carboxyl group or a carboxylate group;

[0022] m is 0, 1 or 2; m is preferably 0; and

[0023] r is independently 0, 1 or 2.

[0024] A preferred embodiment of the present invention provides a compound of formula (I) or its stereoisomers, tautomers or pharmaceutically acceptable salts thereof, which is a compound of formula (II) or its stereoisomers, tautomers or pharmaceutically acceptable salts thereof:

[0025] Ring C, R 1 、R 2 , W and m are as defined in the general formula (I).

[0026] A preferred embodiment of the present invention provides a compound of formula (I) or (II) or its stereoisomers, tautomers or pharmaceutically acceptable salts, wherein W is N.

[0027] A preferred embodiment of the present invention provides a compound of formula (I) or (II) or a stereoisomer, tautomer or a pharmaceutically acceptable salt thereof, wherein ring C is selected from:

[0028] A preferred embodiment of the present invention provides a compound of formula (I) or (II) or its stereoisomers, tautomers or pharmaceutically acceptable salts, wherein:

[0029] R 1 Selected from hydrogen atom, hydroxyl group, alkyl group, cycloalkyl group, heterocyclic group, -C(O)R 3 、-C(O)NR 6 R 7 or -S(O) r R 4 wherein the alkyl or cycloalkyl group is optionally further substituted by one or more R B replaced by;

[0030] R B Each is independently selected from cyano, hydroxy, heterocyclic, aryl, heteroaryl, -OR 5 、-C(O)OR 5 、-NR 6 R 7 、-C(O)NR 6 R 7 or -S(O) r R 5 wherein the heterocyclic or heteroaryl groups are optionally further substituted by one or more groups selected from hydroxy, cyano, alkoxy, haloalkyl, =O, -C(O)OR 8 、-C(O)NR 9 R 10 substituted by a substituent;

[0031] Or, two R B Form a -C(=O)- with the same carbon atom to which it is attached;

[0032] R 3 is selected from hydrogen atom, alkyl, cycloalkyl, heteroaryl or heterocyclic group; wherein the alkyl, cycloalkyl, heteroaryl or heterocyclic group is optionally further substituted by one or more selected from hydroxyl, alkoxy, cyano, heteroaryl, =O, -NR 9 R 10 、-C(O)OR 8 or -C(O)NR 9 R 10 substituted by a substituent;

[0033] R 4 is a hydrogen atom or a hydroxyl group;

[0034] R 5 are each independently selected from a hydrogen atom, an alkyl group or a hydroxyalkyl group;

[0035] R 6 、R 7 are each independently selected from a hydrogen atom or an alkyl group, wherein the alkyl group is optionally substituted with one or more substituents selected from hydroxyl, cyano, halogen or alkoxy;

[0036] R 8 、R 9 、R 10 Each is independently selected from a hydrogen atom or an alkyl group; the alkyl group is preferably a methyl group;

[0037] r is 2.

[0038] In a preferred embodiment of the present invention, the compound described by general formula (I) is selected from:

[0039] or a stereoisomer, a tautomer or a pharmaceutically acceptable salt thereof.

[0040] Note: If there is a discrepancy between a drawn structure and the name given for that structure, the drawn structure will be given greater weight.

[0041] Furthermore, the present invention provides a pharmaceutical composition comprising an effective dose of a compound of formula (I) or (II) or its stereoisomers, tautomers or pharmaceutically acceptable salts, and a pharmaceutically acceptable carrier, excipient or a combination thereof.

[0042] The present invention provides a use of a compound of formula (I) or (II) or its stereoisomers, tautomers or pharmaceutically acceptable salts, or a pharmaceutical composition thereof (such as the pharmaceutical composition described in the aforementioned technical solution) in the preparation of an LPXC inhibitor.

[0043] The present invention also provides a use of a compound of formula (I) or (II) or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof (e.g., the pharmaceutical composition described in the aforementioned technical solution) in the preparation of a medicament for treating a disease mediated by LPXC, wherein the disease mediated by LPXC is preferably a bacterial infection caused by Gram-negative bacteria; wherein the disease mediated by LPXC is more preferably selected from bacterial infections caused by Gram-negative bacteria such as Escherichia coli, Pseudomonas aeruginosa, Proteus, Shigella dysenteriae, Klebsiella pneumoniae, Brucella, Salmonella typhi, Acinetobacter, Yersinia, Legionella pneumophila, Bordetella pertussis, Shigella, Pasteurella, Vibrio cholerae, and Neisseria meningitidis.

[0044] The present invention further provides a use of a compound of general formula (I) or (II) or its stereoisomers, tautomers or pharmaceutically acceptable salts, or a pharmaceutical composition thereof (such as the pharmaceutical composition described in the aforementioned technical solution) in the preparation of a drug for treating bacterial infections caused by Gram-negative bacteria.

[0045] The present invention provides use of a compound of general formula (I) or (II) or its stereoisomers, tautomers or pharmaceutically acceptable salts, or a pharmaceutical composition thereof (e.g., the pharmaceutical composition described in the aforementioned technical solution) in the preparation of a drug for treating bacterial infection, wherein the bacterial infection is caused by Gram-negative bacteria such as Escherichia coli, Pseudomonas aeruginosa, Proteus, Shigella dysenteriae, Klebsiella pneumoniae, Brucella, Salmonella typhi, Acinetobacter, Yersinia, Legionella pneumophila, Bordetella pertussis, Shigella, Pasteurella, Vibrio cholerae, and Neisseria meningitidis.

[0046] The present invention provides a method for preventing and / or treating a disease mediated by LPXC, comprising administering to a patient a therapeutically effective dose of a compound of formula (I) or (II) or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof (e.g., the pharmaceutical composition described in the aforementioned technical solution), wherein the disease mediated by LPXC is preferably a bacterial infection caused by Gram-negative bacteria; wherein the disease mediated by LPXC is more preferably selected from bacterial infections caused by Gram-negative bacteria such as Escherichia coli, Pseudomonas aeruginosa, Proteus, Shigella dysenteriae, Klebsiella pneumoniae, Brucella, Salmonella typhi, Acinetobacter, Yersinia, Legionella pneumophila, Bordetella pertussis, Shigella, Pasteurella, Vibrio cholerae, and Neisseria meningitidis.

[0047] The present invention also provides a method for preventing and / or treating bacterial infections caused by Gram-negative bacteria, comprising administering to a patient a therapeutically effective dose of a compound of formula (I) or (II) or its stereoisomers, tautomers or pharmaceutically acceptable salts, or a pharmaceutical composition thereof (e.g., the pharmaceutical composition described in the aforementioned technical solution).

[0048] The present invention further provides a method for preventing and / or treating bacterial infections caused by Gram-negative bacteria such as Escherichia coli, Pseudomonas aeruginosa, Proteus, Shigella dysenteriae, Klebsiella pneumoniae, Brucella, Salmonella typhi, Acinetobacter, Yersinia, Legionella pneumophila, Bordetella pertussis, Shigella, Pasteurella, Vibrio cholerae, and Neisseria meningitidis, comprising administering to a patient a therapeutically effective dose of a compound of formula (I) or (II) or its stereoisomers, tautomers, or pharmaceutically acceptable salts, or a pharmaceutical composition thereof (e.g., the pharmaceutical composition described in the aforementioned technical solution).

[0049] Detailed Description of the Invention

[0050] Unless otherwise stated, some of the terms used in the specification and claims of the present invention are defined as follows:

[0051] "Alkyl" when used as a group or a part of a group refers to a group comprising C1-C 20 A straight chain or branched aliphatic hydrocarbon group. Preferably C1-C 10 Alkyl, more preferably C1-C6 alkyl. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, etc. Alkyl can be substituted or unsubstituted.

[0052] "Cycloalkyl" refers to a non-aromatic cyclic alkyl group in which one or more of the ring atoms is a carbon atom, including monocyclic, polycyclic, fused, bridged, and spirocyclic rings, preferably having a 5- to 7-membered monocyclic ring or a 7- to 10-membered bicyclic or tricyclic ring. Examples of "cycloalkyl" include, but are not limited to, cyclopropyl, cyclopentyl, and cyclobutyl. Cycloalkyl groups may be substituted or unsubstituted.

[0053] "Spiroalkyl" refers to a polycyclic group having 5 to 18 members, two or more cyclic structures, and a carbon atom (called a spiro atom) shared between the monocyclic rings, containing one or more double bonds in the ring, but no ring has a completely conjugated π electron aromatic system. Preferably, it is 6 to 14 members, more preferably 7 to 10 members. According to the number of spiro atoms shared between the rings, the spiroalkyl is divided into a single spiro, a double spiro or a multi-spiroalkyl, preferably a single spiro and a double spiroalkyl, preferably 4 / 5 members, 4 / 6 members, 5 / 5 members or 5 / 6 members. Non-limiting examples of "spiroalkyl" include, but are not limited to, spiro[4.5]decyl, spiro[4.4]nonyl, spiro[3.5]nonyl, spiro[2.4]heptyl.

[0054] "Fused cycloalkyl" refers to a 5- to 18-membered, all-carbon polycyclic group containing two or more cyclic structures sharing a pair of carbon atoms. One or more rings may contain one or more double bonds, but no ring has a completely conjugated π electron aromatic system. It is preferably 6- to 12-membered, and more preferably 7- to 10-membered. Depending on the number of constituent rings, it can be classified as a bicyclic, tricyclic, tetracyclic or polycyclic fused cycloalkyl, preferably a bicyclic or tricyclic, and more preferably a 5-membered / 5-membered or 5-membered / 6-membered bicyclic alkyl. Non-limiting examples of "fused cycloalkyl" include, but are not limited to: bicyclo[3.1.0]hexyl, bicyclo[3.2.0]hept-1-enyl, bicyclo[3.2.0]heptyl, decahydronaphthyl or tetradecahydrophenanthrenyl.

[0055] "Bridged cycloalkyl" refers to an all-carbon polycyclic group with 5 to 18 members, containing two or more cyclic structures that share two non-directly connected carbon atoms. One or more rings may contain one or more double bonds, but none of the rings have completely conjugated π electrons. It is an aromatic system with preferably 6 to 12 members, more preferably 7 to 10 members. It is preferably 6 to 14 members, more preferably 7 to 10 members. Depending on the number of constituent rings, bridged cycloalkyl groups can be classified as bicyclic, tricyclic, tetracyclic, or polycyclic, preferably bicyclic, tricyclic, or tetracyclic, and more preferably bicyclic or tricyclic. Non-limiting examples of "bridged cycloalkyl" include, but are not limited to: (1s,4s)-bicyclo[2.2.1]heptyl, bicyclo[3.2.1]octyl, (1s,5s)-bicyclo[3.3.1]nonyl, bicyclo[2.2.2]octyl, and (1r,5r)-bicyclo[3.3.2]decyl.

[0056] "Heterocyclyl", "heterocycloalkyl", "heterocycle" or "heterocyclic" are used interchangeably in this application and refer to non-aromatic heterocyclic groups, wherein one or more atoms forming the ring are selected from nitrogen, oxygen or S(O)r (wherein r is selected from 0, 1 or 2) heteroatoms, including monocyclic, polycyclic, fused, bridged and spirocyclic groups, and the heterocyclic group can be substituted or unsubstituted.

[0057] Examples of "monocyclic heterocyclyl" include, but are not limited to, morpholinyl, oxetanyl, thiomorpholinyl, tetrahydrofuranyl, tetrahydropyranyl, 1,1-dioxo-thiomorpholinyl, piperidinyl, 2-oxo-piperidinyl, pyrrolidinyl, 2-oxo-pyrrolidinyl, piperazin-2-one, 8-oxa-3-aza-bicyclo[3.2.1]octyl, piperazinyl, hexahydropyrimidine, 1,4-dioxane, or

[0058] "Spiro heterocyclyl" refers to a polycyclic group with 5 to 18 members, two or more ring structures, and one atom shared between the rings, containing one or more double bonds in the ring, but no ring has a completely conjugated π electron aromatic system, wherein one or more ring atoms are selected from nitrogen, oxygen or S(O) r (wherein r is selected from 0, 1 or 2) heteroatoms, and the remaining ring atoms are carbon. Preferably 5 to 14 members, more preferably 6 to 10 members, most preferably 7 to 10 members. According to the number of shared spiro atoms between rings, spiroalkyl is divided into monospiro heterocyclic group, bispiro heterocyclic group or polyspiro heterocyclic group, preferably monospiro heterocyclic group and bispiro heterocyclic group. More preferably, it is 4 yuan / 4 yuan, 4 yuan / 5 yuan, 4 yuan / 6 yuan, 5 yuan / 5 yuan or 5 yuan / 6 yuan monospiro heterocyclic group. Non-limiting examples of "spiro heterocyclic group" include but are not limited to: 1,7-dioxaspiro[4.5]decyl, 2-oxa-7-azaspiro[4.4]nonyl, 7-oxaspiro[3.5]nonyl, 5-oxaspiro[2.4]heptyl,

[0059] "Fused heterocyclic group" refers to an all-carbon polycyclic group containing two or more ring structures sharing a pair of atoms, one or more rings may contain one or more double bonds, but no ring has a completely conjugated π-electron aromatic system, wherein one or more ring atoms are selected from nitrogen, oxygen or S(O) r (wherein r is selected from 0, 1 or 2) heteroatom, and the remaining ring atoms are carbon. Preferably it is 6 to 14 members, more preferably 7 to 10 members. According to the number of constituent rings, it can be divided into bicyclic, tricyclic, tetracyclic or polycyclic fused heterocyclic groups, preferably bicyclic or tricyclic, more preferably 5-membered / 5-membered or 5-membered / 6-membered bicyclic fused heterocyclic groups. Non-limiting examples of "fused heterocyclic groups" include, but are not limited to: octahydropyrrolo[3,4-c]pyrrolyl, octahydro-1H-isoindolyl, 3-azabicyclo[3.1.0]hexyl, octahydrobenzo[b][1,4]dioxin.

[0060] "Bridged heterocyclic group" refers to a 5- to 14-membered, 5- to 18-membered polycyclic group containing two or more ring structures that share two atoms that are not directly connected to each other, one or more rings may contain one or more double bonds, but no ring has a completely conjugated π-electron aromatic system, wherein one or more ring atoms are selected from nitrogen, oxygen or S(O) r (wherein r is selected from 0, 1 or 2) heteroatoms, and the remaining ring atoms are carbon. Preferably 6 to 14 members, more preferably 7 to 10 members. According to the number of constituent rings, it can be divided into bicyclic, tricyclic, tetracyclic or polycyclic bridged heterocyclic groups, preferably bicyclic, tricyclic or tetracyclic, more preferably bicyclic or tricyclic. Non-limiting examples of "bridged heterocyclic groups" include but are not limited to: 2-azabicyclo[2.2.1]heptyl, 2-azabicyclo[2.2.2]octyl, 2-azabicyclo[3.3.2]decyl.

[0061] "Aryl" refers to a carbocyclic aromatic system containing one or two rings, wherein the rings may be joined together in a fused manner. The term "aryl" includes monocyclic or bicyclic aromatic groups, such as phenyl, naphthyl, tetrahydronaphthyl aromatic groups. Preferably, aryl is C6-C 10 The aryl group is more preferably phenyl and naphthyl, and most preferably naphthyl. The aryl group may be substituted or unsubstituted.

[0062] "Heteroaryl" refers to an aromatic 5- to 6-membered monocyclic or 8- to 10-membered bicyclic ring which may contain 1 to 4 atoms selected from nitrogen, oxygen, and / or sulfur. Examples of "heteroaryl" include, but are not limited to, furanyl, pyridyl, 2-oxo-1,2-dihydropyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, thienyl, isoxazolyl, oxazolyl, oxadiazolyl, imidazolyl, pyrrolyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl, isothiazolyl, 1,2,3-thiadiazolyl, benzodioxolyl, benzothienyl, benzimidazolyl, indolyl, isoindolyl, 1,3-dioxo-isoindolyl, quinolinyl, indazolyl, benzisothiazolyl, benzoxazolyl, benzisoxazolyl, isothiazolyl, 1H-1,2,4-triazolyl, 4H-1,2,4 -triazolyl, pyridyl, pyrimidinyl, pyrazin-2(1H)-onyl, pyrimidin-4(3H)-onyl, pyridazin-3(2H)-onyl, 1H-indolyl, 1H-benzo[d]imidazolyl, 1H-pyrrolo[2,3-c]pyridinyl, 3H-imidazo[4,5-c]pyridinyl, isoquinolinyl, quinazolinyl, 2H-isoindolyl, furo[3,2-b]pyridinyl, furo[2,3-c]pyridinyl, thieno[2,3-c]pyridinyl, benzofuranyl, benzo[b]thienyl, 1H-pyrrolo[3,2-b]pyridinyl, 2H-pyrrolo[3,4-c]pyridinyl. The heteroaryl group may be substituted or unsubstituted.

[0063] "Alkoxy" refers to a group (alkyl-O-). Alkyl is defined herein. C1-C6 alkoxy groups are preferred. Examples include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, and tert-butoxy.

[0064] "Nitro" refers to a -NO2 group.

[0065] "Hydroxy" refers to an -OH group.

[0066] "Halogen" refers to fluorine, chlorine, bromine and iodine.

[0067] "Amino" refers to -NH2.

[0068] "Cyano" refers to -CN.

[0069] "Benzyl" refers to -CH2-phenyl.

[0070] "Carboxyl" refers to -C(O)OH.

[0071] "Carboxylate" refers to a -C(O)O-alkyl group or a -C(O)O-cycloalkyl group, wherein alkyl and cycloalkyl are as defined above.

[0072] "Hydroxyalkyl" or "hydroxyalkyl" refers to an alkyl group substituted with a hydroxy group, wherein alkyl is as defined above.

[0073] "Aminoalkyl" refers to an alkyl group substituted with an amino group, wherein alkyl is as defined above.

[0074] "Haloalkyl" refers to an alkyl group substituted with a halogen, wherein alkyl is as defined above.

[0075] "Haloalkoxy" refers to an alkoxy group substituted with a halogen group, wherein alkoxy is as defined above.

[0076] "DMSO" refers to dimethyl sulfoxide.

[0077] "BOC" refers to tert-butoxycarbonyl.

[0078] "Bn" refers to benzyl.

[0079] "THP" refers to 2-tetrahydropyranyl.

[0080] "TFA" refers to trifluoroacetic acid.

[0081] "Ts" refers to p-toluenesulfonyl.

[0082] "Leaving group", or leaving group, is an atom or functional group that breaks away from a larger molecule in a chemical reaction. It is a term used in nucleophilic substitution reactions and elimination reactions. In a nucleophilic substitution reaction, the reactant attacked by the nucleophile is called the substrate, and the atom or group of atoms that breaks away from the substrate molecule with a pair of electrons is called the leaving group. Groups that easily accept electrons and have a strong ability to withstand negative charges are good leaving groups. The smaller the pKa of the conjugate acid of the leaving group, the easier it is for the leaving group to break away from other molecules. The reason is that when the pKa of its conjugate acid is smaller, the corresponding leaving group does not need to bind to other atoms, and the tendency to exist as an anion (or an electrically neutral leaving group) is enhanced. Common leaving groups include but are not limited to halogens, methylsulfonyl, -OTs or -OH.

[0083] "Substituted" means that one or more hydrogen atoms, preferably up to 5, more preferably 1 to 3 hydrogen atoms, in a group are replaced independently of one another by a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and a person skilled in the art can determine (by experiment or theory) which substitutions are possible or impossible without undue effort. For example, an amino or hydroxyl group with free hydrogen may be unstable when combined with a carbon atom with an unsaturated (e.g., olefinic) bond.

[0084] As used herein, "substituted" or "substituted", unless otherwise specified, means that a group may be substituted by one or more groups selected from the following groups: alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, amino, haloalkyl, hydroxyalkyl, carboxyl, carboxylate, =O, -OR 5 、-C(O)R 5 、-C(O)OR 5 、-NHC(O)R 5 、-NHC(O)OR 5 、-NR 6 R 7 、-C(O)NR 6 R 7 、-CH2NHC(O)OR 5 、-CH2NR 6 R 7 or -S(O)rR 5 substituted by a substituent;

[0085] R 6is selected from hydrogen atom, alkyl, cycloalkyl, heterocyclic, aryl or heteroaryl, wherein said alkyl, cycloalkyl, heterocyclic, aryl or heteroaryl is optionally further substituted by one or more selected from hydroxyl, halogen, nitro, cyano, alkyl, alkoxy, haloalkyl, haloalkoxy, cycloalkyl, heterocyclic, aryl, heteroaryl, =O, -C(O)R 8 、-C(O)OR 8 、-OC(O)R 8 、-NR 9 R 10 、-C(O)NR 9 R 10 、-SO2NR 9 R 10 or -NR 9 C(O)R 10 substituted by a substituent;

[0086] R 6 and R 7 Each is independently selected from hydrogen, hydroxy, halogen, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl or heteroaryl, wherein the alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl or heteroaryl is optionally further substituted by one or more selected from hydroxy, halogen, nitro, cyano, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, =O, -C(O)R 8 、-C(O)OR 8 、-OC(O)R 8 、-NR 9 R 10 、-C(O)NR 9 R 10 、-SO2NR 9 R 10 or -NR 9 C(O)R 10 substituted by a substituent;

[0087] Or, R 6 and R 7 Together with the atoms to which they are attached, they form a 4-8 membered heterocyclic group, wherein the 4-8 membered heterocyclic group contains one or more N, O or S(O)r, and the 4-8 membered heterocyclic group is optionally further substituted by one or more selected from hydroxy, halogen, nitro, cyano, alkyl, alkoxy, cycloalkyl, heterocyclic group, aryl, heteroaryl, =O, -C(O)R 8 、-C(O)OR 8 、-OC(O)R 8 、-NR 9 R 10 、-C(O)NR 9 R 10 、-SO2NR9 R 10 or -NR 9 C(O)R 10 substituted by a substituent;

[0088] R 8 、R 9 and R 10 Each is independently selected from a hydrogen atom, an alkyl group, an amino group, a cycloalkyl group, a heterocyclic group, an aryl group or a heteroaryl group, wherein the alkyl group, the cycloalkyl group, the heterocyclic group, the aryl group or the heteroaryl group is optionally further substituted with one or more substituents selected from a hydroxyl group, a halogen group, a nitro group, an amino group, a cyano group, an alkyl group, an alkoxy group, a cycloalkyl group, a heterocyclic group, an aryl group, a heteroaryl group, a carboxyl group or a carboxylate group;

[0089] r is selected from 0, 1 or 2.

[0090] The compounds of the present invention may contain asymmetric centers or chiral centers and therefore exist in different stereoisomeric forms. It is contemplated that all stereoisomeric forms of the compounds of the present invention, including but not limited to diastereomers, enantiomers and atropisomers and geometric (conformational) isomers and mixtures thereof, such as racemic mixtures, are within the scope of the present invention.

[0091] Unless otherwise indicated, structures depicted herein also encompass all isomers (e.g., diastereoisomers, enantiomers, and atropisomers, and geometric (conformational) isomeric forms of such structures; for example, R and S configurations at various asymmetric centers, (Z) and (E) double bond isomers, and (Z) and (E) conformational isomers. Therefore, individual stereoisomers as well as enantiomeric mixtures, diastereomeric mixtures, and geometric (conformational) isomeric mixtures of the present compounds are within the scope of the invention.

[0092] "Pharmaceutically acceptable salts" refer to salts of the above compounds that retain their original biological activity and are suitable for pharmaceutical use. Pharmaceutically acceptable salts of the compounds represented by general formula (I) may be metal salts or amine salts formed with suitable acids.

[0093] A "pharmaceutical composition" refers to a mixture containing one or more compounds described herein, or their physiologically acceptable salts or prodrugs, together with other chemical components, as well as other components such as physiologically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration to an organism, facilitating absorption of the active ingredients and thereby exerting their biological activity.

[0094] Synthesis method of the compound of the present invention

[0095] In order to achieve the purpose of the present invention, the present invention adopts the following technical solutions:

[0096] The present invention provides a method for preparing a compound of general formula (I) or its stereoisomers, tautomers or pharmaceutically acceptable salts, the method comprising:

[0097] When W is N:

[0098] Method 1:

[0099] The compound of general formula (Ia) and the compound of general formula (Ib) undergo a coupling reaction in the presence of a catalyst, and optionally further undergo a deprotection reaction to obtain a compound of general formula (Ic). The compound of general formula (Ic) optionally further undergoes a substitution reaction with a compound of general formula (Id) to obtain a compound of general formula (I).

[0100] in:

[0101] X1 and X2 are selected from halogen;

[0102] PG is an amino protecting group, preferably tert-butyloxycarbonyl;

[0103] PG' is a hydroxyl protecting group, preferably 2-tetrahydropyranyl;

[0104] Ring C, X, Y, Z, Q, R 1 、R 2 and m are as described in general formula (I).

[0105] Method 2:

[0106] The compound of general formula (Ia) undergoes a coupling reaction with the compound of general formula (II-b) in the presence of a catalyst to obtain a compound of general formula (II-c). The compound of general formula (II-c) further undergoes a deprotection reaction to obtain a compound of general formula (I).

[0107] in:

[0108] X1 is selected from halogen;

[0109] PG' is a hydroxyl protecting group, preferably 2-tetrahydropyranyl;

[0110] Ring C, X, Y, Z, Q, R 1 、R 2 and m are as described in general formula (I). DETAILED DESCRIPTION

[0111] The present invention is further described below with reference to the following examples, but these examples are not intended to limit the scope of the present invention.

[0112] Example

[0113] The examples provide the preparation of representative compounds represented by formula (I) and related structural identification data. It must be noted that the following examples are used to illustrate the present invention rather than to limit the present invention. 1 H NMR spectra were obtained using a Bruker instrument (400 MHz), and chemical shifts are expressed in ppm using tetramethylsilane as an internal standard (0.00 ppm). 1 H NMR notation: s = singlet, d = doublet, t = triplet, m = multiplet, br = broadened, dd = double of doublets, dt = double of triplets, dq = double of quartets. Coupling constants, when given, are in Hz.

[0114] Mass spectra were obtained using LC / MS, and the ionization method could be ESI or APCI.

[0115] The thin layer chromatography silica gel plate uses Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plate. The specification of the silica gel plate used in thin layer chromatography (TLC) is 0.15mm~0.2mm, and the specification used for thin layer chromatography separation and purification products is 0.4mm~0.5mm.

[0116] Column chromatography generally uses Yantai Huanghai silica gel 200-300 mesh silica gel as the carrier.

[0117] In the following examples, unless otherwise indicated, all temperatures are in degrees Celsius. Unless otherwise indicated, various starting materials and reagents are commercially available or synthesized according to known methods. Commercially available raw materials and reagents are used directly without further purification, unless otherwise indicated. Commercial manufacturers include but are not limited to Aldrich Chemical Company, ABCR GmbH & Co. KG, Acros Organics, Guangzan Chemical Technology Co., Ltd. and Jingyan Chemical Technology Co., Ltd.

[0118] DMSO-d6: deuterated dimethyl sulfoxide.

[0119] Argon atmosphere means that the reaction bottle is connected to an argon balloon with a capacity of about 1 L.

[0120] Unless otherwise specified in the examples, the solution in the reaction refers to an aqueous solution.

[0121] The compound is purified using a silica gel column chromatography eluent system and thin layer chromatography, wherein the eluent system is selected from: A: petroleum ether and ethyl acetate system; B: dichloromethane and methanol system; C: dichloromethane and ethyl acetate system; wherein the volume ratio of the solvent varies according to the polarity of the compound and can also be adjusted by adding a small amount of acidic or alkaline reagents, such as acetic acid or triethylamine.

[0122] Example 1

[0123] (S)-1-(1-((5-(4-((7-azaspiro[3.5]nonan-2-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol

[0124] (S)-1-(1-((5-(4-((7-Azaspiro[3.5]non-2-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol

[0125] first step

[0126] Ethyl 4-(4-iodophenyl)-2,4-dioxobutanoate

[0127] Sodium hydride (1.63 g, 40.64 mmol, 60% oil dispersion) was added to toluene (20 mL), followed by 1-(4-iodophenyl)ethan-1-one 1a (5 g, 20.32 mmol). The mixture was heated to 50°C, and a toluene solution (20 mL) of diethyl oxalate 1b (4.45 g, 30.48 mmol) was added dropwise. The mixture was heated to 50°C and allowed to react for 2 hours. The mixture was cooled, poured into ice water, and acidified with 1M hydrochloric acid. The mixture was extracted with ethyl acetate (100 mL x 2). The combined organic phases were washed with saturated brine (100 mL x 3), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: System A) to give ethyl 4-(4-iodophenyl)-2,4-dioxobutanoate 1c (2.8 g) in a yield of 39.81%.

[0128] MS m / z(ESI):347.0[M+1].

[0129] Step 2

[0130] Ethyl 5-(4-iodophenyl)isoxazole-3-carboxylate

[0131] Ethyl 4-(4-iodophenyl)-2,4-dioxobutanoate 1c (2.8 g, 8.09 mmol) and hydroxylamine hydrochloride (1.69 g, 24.27 mmol) were added to ethanol (25 mL) and heated under reflux for 1 hour. After completion of the reaction, the mixture was concentrated under reduced pressure and dissolved in ethyl acetate (100 mL). The organic phase was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (eluent: System A) to give ethyl 5-(4-iodophenyl)isoxazole-3-carboxylate 1d (2.3 g) in a yield of 82.86%.

[0132] MS m / z(ESI):343.8[M+1].

[0133] Step 3

[0134] (5-(4-iodophenyl)isoxazol-3-yl)methanol

[0135] Ethyl 5-(4-iodophenyl)isoxazole-3-carboxylate 1d (1.20 g, 3.50 mmol) was added to methanol (25 mL), and sodium borohydride (198.46 mg, 5.25 mmol) was added portionwise. The temperature was raised to 80°C and the reaction was allowed to react for 4 hours. After completion of the reaction, ice water was added to quench the reaction, and the mixture was extracted with ethyl acetate (50 mL x 2). The organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to provide (5-(4-iodophenyl)isoxazol-3-yl)methanol 1e (0.36 g) in a yield of 34.19%.

[0136] MS m / z(ESI):302.0[M+1].

[0137] Step 4

[0138] Methyl (5-(4-iodophenyl)isoxazol-3-yl)methanesulfonate

[0139] (5-(4-iodophenyl)isoxazol-3-yl)methanol 1e (0.36 g, 1.20 mmol) and triethylamine (241.99 mg, 2.39 mmol, 333.31 μL) were added to dichloromethane (5 mL). The mixture was cooled to 0°C, and methanesulfonyl chloride 1f (205.45 mg, 1.79 mmol) was added dropwise. The mixture was allowed to warm to room temperature and allowed to react for 4 hours. After completion, the reaction was quenched with water and extracted with dichloromethane (50 mL x 3). The organic phases were combined and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (eluent: System A) to obtain methyl (5-(4-iodophenyl)isoxazol-3-yl)methanesulfonate 1g (0.45 g) in a yield of 99.26%.

[0140] MS m / z(ESI):379.8[M+1].

[0141] Step 5

[0142] 5-(4-iodophenyl)-3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazole

[0143] 2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazole 1h (100 mg, 509.57 μmol) was added to N,N-dimethylformamide (2 mL), cooled to 0°C, sodium hydride (50.96 mg, 764.35 μmol, 60% oil dispersion) was added portionwise, and the mixture was heated to room temperature for 1 hour. 1 g (193.21 mg, 509.57 μmol) of methyl (5-(4-iodophenyl)isoxazol-3-yl)methanesulfonate was added, and the reaction was continued at room temperature for 4 hours. After the reaction was completed, water was added to quench the reaction, and the product was extracted with ethyl acetate (10 mL × 3). The organic phases were combined and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: System A) to give 5-(4-iodophenyl)-3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazole 1i (0.12 g) in a yield of 49.13%.

[0144] MS m / z(ESI):480.1[M+1].

[0145] Step 6

[0146] tert-butyl 2-ethynyl-7-azaspiro[3.5]nonane-7-carboxylate

[0147] tert-Butyl 2-ethynyl-7-azaspiro[3.5]nonane-7-carboxylate

[0148] Dimethyl (1-diazo-2-oxopropyl)phosphonate (1.36 g, 7.11 mmol) was added to a solution of potassium carbonate (1.64 g, 11.8 mmol) and tert-butyl 2-formyl-7-azaspiro[3.5]nonane-7-carboxylate 1j (1.5 g, 5.92 mmol, commercially available) in methanol (15 mL) under ice-cooling and allowed to react at room temperature for 24 hours. After completion, the reaction was quenched by the addition of water (50 mL), extracted with ethyl acetate (50 mL × 3), and the combined organic phases were washed with saturated brine (100 mL) and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (eluent: System A) to afford tert-butyl 2-ethynyl-7-azaspiro[3.5]nonane-7-carboxylate 1k (1.3 g) in an 88.1% yield.

[0149] MS m / z(ESI):194.1[M+1-56].

[0150] Step 7

[0151] tert-butyl

[0152] 2-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonane-7-carboxylate

[0153] tert-Butyl 2-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonane-7-carboxylate

[0154] 5-(4-iodophenyl)-3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazole 1i (1.3 g, 2.71 mmol), tert-butyl 2-ethynyl-7-azaspiro[3.5]nonane-7-carboxylate 1k (1.26 g, 3.53 mmol), bistriphenylphosphine palladium dichloride (190 mg, 271 μmol), cuprous iodide (51.8 mg, 271 μmol) and triethylamine (823 mg, 8.14 mmol, 1.13 mL) were added to N,N-dimethylformamide (13.87 mL), the argon atmosphere was replaced three times, and the reaction was carried out at room temperature overnight. After completion of the reaction, water (30 mL) was added to quench the reaction, and the mixture was extracted with ethyl acetate (100 mL×3). The combined organic phases were washed with saturated brine (100 mL) and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: System B) to give tert-butyl 2-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonane-7-carboxylate 11 (1.25 g) in a yield of 76.7%.

[0155] MS m / z(ESI):601.1[M+1].

[0156] Step 8

[0157] (S)-1-(1-((5-(4-((7-azaspiro[3.5]nonan-2-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol

[0158] (S)-1-(1-((5-(4-((7-Azaspiro[3.5]non-2-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol

[0159] Tert-butyl 2-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonane-7-carboxylate 11 (1.2 g, 2.00 mmol) was dissolved in dichloromethane (30 mL), trifluoroacetic acid (228 mg, 2.00 mmol) was slowly added, and the reaction was carried out at room temperature for 4 hours. After the reaction was completed, the mixture was concentrated and subjected to preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm ID; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H2O, mobile phase B: CH3CN) to give (S)-1-(1-((5-(4-((7-azaspiro[3.5]non-2-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol 1 (0.85 g) in a yield of 76.2%.

[0160] MS m / z(ESI):417.1[M+1].

[0161] Example 2

[0162] (S)-4-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)butanoic acid

[0163] (S)-4-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)butanoic acid

[0164] first step

[0165] ethyl

[0166] (S)-4-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)butanoate

[0167] (S)-ethyl 4-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)butanoate

[0168] (S)-1-(1-((5-(4-((7-azaspiro[3.5]non-2-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol 1 (100 mg, 240 μmol), ethyl 4-bromobutyrate 2a (70.3 mg, 360 μmol, commercially available), potassium carbonate (332 mg, 2.40 mmol) and sodium iodide (119 mg, 792 μmol) were added sequentially to N,N-dimethylformamide (3 mL) and heated to 70 °C for 4 h. After completion of the reaction, water (30 mL) was added to quench the reaction, and the mixture was extracted with ethyl acetate (50 mL×3). The combined organic phases were washed with saturated brine (100 mL) and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: System B) to give (S)-ethyl 4-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]non-7-yl)butanoate 2b (65 mg) in a yield of 51.0%.

[0169] MS m / z(ESI):531.1[M+1].

[0170] Step 2

[0171] (S)-4-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)butanoic acid

[0172] (S)-4-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)butanoic acid

[0173] Ethyl (S)-4-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]non-7-yl)butanoate 2b (30 mg, 56.5 μmol) was dissolved in a mixed solvent of methanol (0.5 mL), tetrahydrofuran (0.5 mL) and water (0.5 mL). Sodium hydroxide (2.26 mg, 56.5 μmol) was added and the mixture was heated to 70°C for 4 hours. After the reaction was completed, a small amount of dilute hydrochloric acid was added to adjust the pH to about 4, and the organic solvent was removed by concentration. Preparative liquid separation was performed (separation column AKZONOBEL Kromasil; 250×21.2 mm ID; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H2O, mobile phase B: CH3CN) to obtain (S)-4-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]non-7-yl)butanoic acid 2 (1.8 mg) in a yield of 4.9%.

[0174] MS m / z(ESI):503.1[M+1].

[0175] Example 3

[0176] (S)-1-(1-((5-(4-((6-azaspiro[3.4]octan-2-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol

[0177] (S)-1-(1-((5-(4-((6-Azaspiro[3.4]octan-2-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol

[0178] first step

[0179] tert-butyl 2-ethynyl-6-azaspiro[3.4]octane-6-carboxylate

[0180] tert-Butyl 2-ethynyl-6-azaspiro[3.4]octane-6-carboxylate

[0181] Under argon protection, dimethyl (1-diazo-2-oxopropyl)phosphonate (241 mg, 1.25 mmol) was added to a solution of potassium carbonate (289 mg, 2.09 mmol) and tert-butyl 2-formyl-6-azaspiro[3.4]octane-6-carboxylate 3a (250 mg, 1.04 mmol) in methanol (4 mL) under ice-cooling. The mixture was allowed to react at room temperature for 24 hours. After completion, the reaction was diluted with water (20 mL) and extracted with ethyl acetate (50 mL × 3). The combined organic phases were washed with saturated brine (50 mL) and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (eluent: System A) to give tert-butyl 2-ethynyl-6-azaspiro[3.4]octane-6-carboxylate 3b (180 mg) in a yield of 73.2%.

[0182] MS m / z(ESI):170.1[M-55].

[0183] Step 2

[0184] tert-butyl

[0185] 2-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-6-azaspiro[3.4]octane-6-carboxylate

[0186] tert-Butyl 2-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-6-azaspiro[3.4]octane-6-carboxylate

[0187] 5-(4-iodophenyl)-3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazole 1i (0.4 g, 835 μmol), tert-butyl 2-ethynyl-6-azaspiro[3.4]octane-6-carboxylate 3b (365 mg, 1.08 mmol), bistriphenylphosphine palladium dichloride (58.5 mg, 83.5 μmol), cuprous iodide (15.9 mg, 83.5 μmol) and triethylamine (253 mg, 2.50 mmol, 348 μL) were mixed with N,N-dimethylformamide (4.65 mL), the argon atmosphere was replaced three times, and the reaction was carried out at room temperature overnight. After completion of the reaction, water (20 mL) was added to quench the reaction, and the mixture was extracted with ethyl acetate (30 mL×3). The combined organic phases were washed with saturated brine (50 mL) and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: System B) to give tert-butyl 2-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-6-azaspiro[3.4]octane-6-carboxylate 3c (0.4 g) in a yield of 81.7%.

[0188] MS m / z(ESI):587.3[M+1].

[0189] Step 3

[0190] (S)-1-(1-((5-(4-((6-azaspiro[3.4]octan-2-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol

[0191] (S)-1-(1-((5-(4-((6-Azaspiro[3.4]octan-2-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol

[0192] Tert-butyl 2-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-6-azaspiro[3.4]octane-6-carboxylate 3c (0.3 g, 511 μmol) was dissolved in dichloromethane (5 mL). Trifluoroacetic acid (58 mg, 511 μmol) was slowly added dropwise and reacted at room temperature for 4 hours. After completion of the reaction, the mixture was concentrated. Preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H2O, mobile phase B: CH3CN) gave (S)-1-(1-((5-(4-((6-azaspiro[3.4]octan-2-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol 3 (120 mg) in a yield of 43.2%.

[0193] MS m / z(ESI):403.1[M+1].

[0194] Example 4

[0195] (S)-4-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-6-azaspiro[3.4]octan-6-yl)butanoic acid

[0196] (S)-4-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-6-azaspiro[3.4]octan-6-yl)butanoic acid

[0197] first step

[0198] ethyl

[0199] (S)-4-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-6-azaspiro[3.4]octan-6-yl)butanoate

[0200] (S)-ethyl 4-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-6-azaspiro[3.4]octan-6-yl)butanoate

[0201] (S)-1-(1-((5-(4-((6-Azaspiro[3.4]octan-2-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol 3 (100 mg, 248 μmol), ethyl 4-bromobutyrate 2a (72.7 mg, 373 μmol), potassium carbonate (343 mg, 2.48 mmol) and sodium iodide (74.5 mg, 497 μmol) were added to N,N-dimethylformamide (3 mL) and heated to 70 °C for 4 hours. After completion of the reaction, water (20 mL) was added to quench the reaction, and the mixture was extracted with ethyl acetate (50 mL×3). The combined organic phases were washed with saturated brine (50 mL) and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: System B) to give (S)-ethyl 4-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-6-azaspiro[3.4]octan-6-yl)butanoate 4a (60 mg) in a yield of 46.7%.

[0202] MS m / z(ESI):517.1[M+1].

[0203] Step 2

[0204] (S)-4-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-6-azaspiro[3.4]octan-6-yl)butanoic acid

[0205] (S)-4-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-6-azaspiro[3.4]octan-6-yl)butanoic acid

[0206] Ethyl (S)-4-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-6-azaspiro[3.4]octan-6-yl)butanoate 4a (60 mg, 116 μmol) was dissolved in a mixture of methanol (1 mL), tetrahydrofuran (1 mL), and water (1 mL). Sodium hydroxide (23 mg, 580 μmol) was added and the mixture was heated to 70°C for 4 hours. After the reaction was complete, a small amount of dilute hydrochloric acid was added to adjust the pH to approximately 4. The reaction mixture was concentrated to remove most of the organic solvent, and preparative liquid separation was performed (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H2O, mobile phase B: CH3CN) to give (S)-4-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-6-azaspiro[3.4]octan-6-yl)butanoic acid 4 (30 mg) in a yield of 40.7%.

[0207] MS m / z(ESI):489.1[M+1].

[0208] Example 5

[0209] (S)-3-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)propanoic acid

[0210] (S)-3-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)propanoic acid

[0211] first step

[0212] methyl

[0213] (S)-3-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)propanoate

[0214] (S)-methyl 3-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)propanoate

[0215] (S)-1-(1-((5-(4-((7-Azaspiro[3.5]non-2-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol 1 (50 mg, 120 μmol), methyl 3-bromopropionate 5a (30 mg, 180 μmol), potassium carbonate (166 mg, 1.20 mmol) and sodium iodide (59.4 mg, 396 μmol) were added to N,N-dimethylformamide (3 mL) and heated to 70 °C for 4 hours. After completion of the reaction, water (20 mL) was added to dilute the reaction, and the mixture was extracted with ethyl acetate (30 mL×3). The combined organic phases were washed with saturated brine (50 mL) and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: System A) to give (S)-methyl 3-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)propanoate 5b (40 mg) in a yield of 66.7%.

[0216] MS m / z(ESI):503.1[M+1].

[0217] Step 2

[0218] (S)-3-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)propanoic acid

[0219] (S)-3-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)propanoic acid

[0220] Methyl (S)-3-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)propanoate 5b (80 mg, 159 μmol) was dissolved in a mixed solvent of methanol (1 mL), tetrahydrofuran (1 mL) and water (1 mL). Sodium hydroxide (6.4 mg, 159 μmol) was added and the mixture was heated to 70°C for 4 hours. After the reaction was completed, a small amount of dilute hydrochloric acid was added to adjust the pH to about 4, and the product was concentrated to remove most of the organic solvent. Preparative liquid separation was performed (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H 2 O, mobile phase B: CH 3 CN) to obtain (S)-3-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]non-7-yl)propanoic acid 5 (20 mg) in a yield of 19.8%.

[0221] MS m / z(ESI):489.1[M+1].

[0222] Example 6

[0223] (S)-1-(1-((5-(4-((2-azaspiro[3.3]heptan-6-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol

[0224] (S)-1-(1-((5-(4-((2-Azaspiro[3.3]hept-6-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol

[0225] first step

[0226] tert-butyl 6-ethynyl-2-azaspiro[3.3]heptane-2-carboxylate

[0227] tert-Butyl 6-ethynyl-2-azaspiro[3.3]heptane-2-carboxylate

[0228] Under argon, dimethyl (1-diazo-2-oxopropyl)phosphonate (614 mg, 3.20 mmol) was slowly added to a solution of potassium carbonate (736 mg, 5.33 mmol) and tert-butyl 6-formyl-2-azaspiro[3.3]heptane-2-carboxylate 6a (0.6 g, 2.66 mmol) in methanol (8 mL) under ice-cooling. The mixture was allowed to react overnight at room temperature. After completion, water (20 mL) was added to dilute the reaction mixture, and the mixture was extracted with ethyl acetate (30 mL × 3). The combined organic phases were washed with saturated brine (50 mL) and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (eluent: System A) to give tert-butyl 6-ethynyl-2-azaspiro[3.3]heptane-2-carboxylate 6b (0.455 g) in a yield of 77.4%.

[0229] MS m / z(ESI):166.1[M-55].

[0230] Step 2

[0231] tert-butyl

[0232] 6-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-2-azaspiro[3.3]heptane-2-carboxylate

[0233] tert-Butyl 6-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-2-azaspiro[3.3]heptane-2-carboxylate

[0234] 5-(4-Iodophenyl)-3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazole 1i (0.6 g, 1.25 mmol), tert-butyl 6-ethynyl-2-azaspiro[3.3]heptane-2-carboxylate 6b (455 mg, 1.44 mmol), bistriphenylphosphine palladium dichloride (87.8 mg, 125 μmol), cuprous iodide (23.9 mg, 125 μmol) and triethylamine (380 mg, 3.76 mmol, 522 μL) were added sequentially to N,N-dimethylformamide (4 mL), the atmosphere was replaced with argon, and the reaction was carried out at room temperature overnight. After completion of the reaction, water (20 mL) was added to dilute the reaction, and the mixture was extracted with ethyl acetate (100 mL×3). The combined organic phases were washed with saturated brine (100 mL) and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: System B) to give tert-butyl 6-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-2-azaspiro[3.3]heptane-2-carboxylate 6c (0.65 g) in a yield of 90.7%.

[0235] MS m / z(ESI):573.1[M+1].

[0236] Step 3

[0237] (S)-1-(1-((5-(4-((2-azaspiro[3.3]heptan-6-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol

[0238] (S)-1-(1-((5-(4-((2-Azaspiro[3.3]hept-6-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol

[0239] Tert-butyl 6-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-2-azaspiro[3.3]heptane-2-carboxylate 6c (20 mg, 34.92 μmol) was dissolved in dichloromethane (1 mL), and trifluoroacetic acid (46 mg, 405 μmol) was slowly added dropwise, and the reaction was carried out at room temperature for 3 hours. After the reaction was completed, the product was concentrated and the residue was subjected to preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H 2 O, mobile phase B: CH 3 CN) to give (S)-1-(1-((5-(4-((2-azaspiro[3.3]hept-6-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol 6 (5 mg) in a yield of 28.5%.

[0240] MS m / z(ESI):389.1[M+1].

[0241] Example 7

[0242] (S)-4-(6-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-2-azaspiro[3.3]heptan-2-yl)butanoic acid

[0243] (S)-4-(6-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-2-azaspiro[3.3]hept-2-yl)butanoic acid

[0244] first step

[0245] ethyl

[0246] (S)-4-(6-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-2-azaspiro[3.3]heptan-2-yl)butanoate

[0247] (S)-ethyl 4-(6-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-2-azaspiro[3.3]hept-2-yl)butanoate

[0248] (S)-1-(1-((5-(4-((2-azaspiro[3.3]hept-6-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol 6 (135 mg, 348 μmol), ethyl 4-bromobutyrate 2a (102 mg, 521 μmol, commercially available), potassium carbonate (480 mg, 3.48 mmol) and sodium iodide (172 mg, 1.15 mmol) were dissolved in N,N-dimethylformamide (3 mL) and heated to 70 °C for 3 h. After completion of the reaction, water (20 mL) was added to dilute the reaction, and the mixture was extracted with ethyl acetate (50 mL×3). The combined organic phases were washed with saturated brine (50 mL) and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: System B) to give (S)-ethyl 4-(6-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-2-azaspiro[3.3]hept-2-yl)butanoate 7a (50 mg) in a yield of 28.6%.

[0249] MS m / z(ESI):503.1[M+1].

[0250] Step 2

[0251] (S)-4-(6-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-2-azaspiro[3.3]heptan-2-yl)butanoic acid

[0252] (S)-4-(6-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-2-azaspiro[3.3]hept-2-yl)butanoic acid

[0253] Ethyl (S)-4-(6-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-2-azaspiro[3.3]hept-2-yl)butanoate 7a (30 mg, 59.69 μmol) was dissolved in a mixed solvent of methanol (0.5 mL), tetrahydrofuran (0.5 mL) and water (0.5 mL). Sodium hydroxide (2.4 mg, 59.7 μmol) was added and the temperature was raised to 70°C for 3 hours. After the reaction was completed, an appropriate amount of dilute hydrochloric acid was added to adjust the pH to about 4. The organic solvent was removed by concentration, and the product was subjected to preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H 2 O, mobile phase B: CH 3 CN) to obtain (S)-4-(6-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-2-azaspiro[3.3]hept-2-yl)butanoic acid 7 (10 mg) in a yield of 27.0%.

[0254] MS m / z(ESI):475.1[M+1].

[0255] Example 8

[0256] (S)-4-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)butanamide

[0257] (S)-4-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)butanamide

[0258] Ethyl (S)-4-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)butanoate 2b (30 mg, 56.53 μmol) was dissolved in ethanol (1.5 mL) and aqueous ammonia (1 mL) was added dropwise. The mixture was reacted at room temperature overnight. After completion, the reaction was concentrated. After preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H2O, mobile phase B: CH3CN), (S)-4-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]non-7-yl)butanamide 8 (5 mg) was obtained with a yield of 13.65%.

[0259] MS m / z(ESI):502.1[M+1].

[0260] Example 9

[0261] (S)-3-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)propanamide

[0262] (S)-3-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)propanamide

[0263] Methyl (S)-3-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)propanoate 5b (60 mg, 119.38 μmol) was dissolved in ethanol (2 mL), and aqueous ammonia (1 mL) was added dropwise. The reaction was allowed to proceed at room temperature overnight. After the reaction was completed, the mixture was concentrated and subjected to preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H2O, mobile phase B: CH3CN) to give (S)-3-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)propanamide 9 (10 mg) in a yield of 7.18%.

[0264] MS m / z(ESI):488.1[M+1].

[0265] Example 10

[0266] (S)-2-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)acetic acid

[0267] (S)-2-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)acetic acid

[0268] first step

[0269] methyl

[0270] (S)-2-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)acetate

[0271] (S)-methyl 2-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)acetate

[0272] (S)-1-(1-((5-(4-((7-azaspiro[3.5]non-2-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol 1 (0.1 g, 240.09 μmol), methyl 2-bromoacetate 10a (55.1 mg, 360.13 μmol, commercially available), potassium carbonate (332 The mixture of 4-[4-[[4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]non-7-yl)acetate 10b (20 mg) was sequentially added to N,N-dimethylformamide (2 mL) and the reaction was carried out at 80°C for 4 hours. After the reaction was basically completed, water (50 mL) was added to quench the reaction. The mixture was extracted three times with ethyl acetate (50 mL×3). The organic phases were combined and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (eluent: System B) to give (S)-methyl 2-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]non-7-yl)acetate 10b (20 mg) in a yield of 17.05%.

[0273] MS m / z(ESI):489.3[M+1].

[0274] Step 2

[0275] (S)-2-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)acetic acid

[0276] (S)-2-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)acetic acid

[0277] Methyl (S)-2-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)acetate 10b (15 mg, 30.70 μmol) was dissolved in a mixed solvent of methanol (0.5 mL), tetrahydrofuran (0.5 mL) and water (0.5 mL). Sodium hydroxide (6.14 mg, 153.51 μmol) was added and the reaction was carried out at 80°C for 8 h. After the reaction was completed, 1 M dilute hydrochloric acid (3 mL) and water (50 ml) were added, and the mixture was extracted with ethyl acetate (50 mL×3). The organic phases were combined and concentrated under reduced pressure. The product was separated by preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H2O, mobile phase B: CH3CN) to give (S)-2-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]non-7-yl)acetic acid 10 (2.90 mg) in a yield of 18.91%.

[0278] MS m / z(ESI):475.0[M+1].

[0279] Example 11

[0280] (S)-2-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)ethan-1-ol

[0281] (S)-2-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)ethan-1-ol

[0282] 2-Bromoethanol 11a (30 mg, 240.07 μmol, commercially available), (S)-1-(1-((5-(4-((7-azaspiro[3.5]non-2-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol 1 (100 mg, 240.07 μmol), sodium iodide (216 mg, 1.44 mmol), and cesium carbonate (78.2 mg, 240.07 μmol) were added to N,N-dimethylformamide (2 mL) respectively. After reacting at 80°C for 6 hours, the reaction was basically completed. The mixture was quenched with water (50 mL), extracted three times with ethyl acetate (50 mL×3), and the combined organic phases were washed with saturated brine (50 mL), concentrated under reduced pressure, and purified by silica gel column chromatography (eluent: System B) to give (S)-2-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]non-7-yl)ethan-1-ol 11 (10 mg) in a yield of 6.89%.

[0283] MS m / z(ESI):461.0[M+1].

[0284] Example 12

[0285] (S)-2-hydroxy-1-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)ethan-1-one

[0286] (S)-2-Hydroxy-1-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)ethan-1-one

[0287] 2-Hydroxyacetic acid 12a (10 mg, 131.49 μmol, commercially available), (S)-1-(1-((5-(4-((7-azaspiro[3.5]non-2-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol 1 (54.8 mg, 131.49 μmol), N,N-diisopropylethylamine (51.0 mg, 394.47 μmol), and O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (100 mg, 262.98 μmol) were added sequentially to N,N-dimethylformamide (1 mL) and reacted at room temperature for 16 hours. After the reaction was substantially completed, water (50 mL) was added to quench the reaction, and the product was extracted with ethyl acetate (50 mL×3) three times. The organic phases were then combined, washed with saturated brine (50 mL), and subjected to preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H2O, mobile phase B: CH3CN) to give (S)-2-hydroxy-1-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]non-7-yl)ethan-1-one 12 (10 mg) in a yield of 12.28%.

[0288] MS m / z(ESI):475.1[M+1].

[0289] Example 13

[0290] (S)-2-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)acetamide

[0291] (S)-2-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)acetamide

[0292] 2-Bromoacetamide 13a (10 mg, 72.48 μmol, commercially available), (S)-1-(1-((5-(4-((7-azaspiro[3.5]non-2-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol 1 (30.2 mg, 72.48 μmol), potassium carbonate (30.1 mg, 217.45 μmol), and sodium iodide (10.9 mg, 72.48 μmol) were added sequentially to N,N-dimethylformamide (1 mL) and reacted at 80°C for 4 hours. After the reaction was substantially completed, water (50 mL) was added to quench the reaction, and the product was extracted with ethyl acetate (50 mL×3 three times). The organic phases were combined, washed with saturated brine (50 mL), and subjected to preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H2O, mobile phase B: CH3CN) to give (S)-2-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]non-7-yl)acetamide 13 (5 mg) in a yield of 13.84%.

[0293] MS m / z(ESI):474.0[M+1].

[0294] Example 14

[0295] (S)-3-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)-3-oxopropanoic acid

[0296] (S)-3-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)-3-oxopropanoic acid

[0297] first step

[0298] methyl

[0299] (S)-3-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)-3-oxopropanoate

[0300] (S)-methyl 3-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)-3-oxopropanoate

[0301] (S)-1-(1-((5-(4-((7-azaspiro[3.5]non-2-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol 1 (100 mg, 240.09 μmol), methyl 3-chloro-3-oxopropanoate 14a (32.8 mg, 240.09 μmol, commercially available), and triethylamine (72.9 mg, 720.26 μmol) were added to N,N-dimethylformamide (3 mL) and reacted at room temperature for 8 hours. After the reaction was substantially completed, the mixture was concentrated and subjected to preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H 2 O, mobile phase B: CH 3 CN) to give (S)-3-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]non-7-yl)-3-oxopropanoic acid methyl ester 14b (40 mg) in a yield of 32.25%.

[0302] MS m / z(ESI):517.0[M+1].

[0303] Step 2

[0304] (S)-3-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)-3-oxopropanoic acid

[0305] (S)-3-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)-3-oxopropanoic acid

[0306] Methyl (S)-3-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)-3-oxopropanoate 14b (5 mg, 9.68 μmol) was dissolved in a mixed solvent of methanol (0.25 mL), tetrahydrofuran (0.25 mL) and water (0.25 mL). Sodium hydroxide (1.16 mg, 29.04 μmol) was added and the reaction was carried out at 80°C for 4 hours. After the reaction was completed, dilute hydrochloric acid (3 mL) and water (50 mL) were added, and the product was extracted with ethyl acetate (50 mL×3). The combined organic phases were washed with saturated brine (50 mL) and subjected to preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H2O, mobile phase B: CH3CN) to give (S)-3-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]non-7-yl)-3-oxopropanoic acid 14 (2.70 mg) in a yield of 52.73%.

[0307] MS m / z(ESI):503.0[M+1].

[0308] Example 15

[0309] (S)-3-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)-3-oxopropanamide

[0310] (S)-3-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)-3-oxopropanamide

[0311] Methyl (S)-3-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)-3-oxopropanoate 14b (20 mg, 38.72 μmol) was dissolved in methanol (1 mL), followed by dropwise addition of aqueous ammonia (0.8 mL) and reaction at room temperature for 36 h. After the reaction was substantially completed, the mixture was concentrated and subjected to preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H2O, mobile phase B: CH3CN) to give (S)-3-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]non-7-yl)-3-oxopropanamide 15 (2.66 mg) in a yield of 10.60%.

[0312] MS m / z(ESI):502.0[M+1].

[0313] Example 16

[0314] (S)-2-amino-1-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)ethan-1-one

[0315] (S)-2-Amino-1-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)ethan-1-one

[0316] first step

[0317] tert-butyl

[0318] (S)-(2-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)-2-oxoethyl)carbamate

[0319] (S)-tert-Butyl(2-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)-2-oxoethyl)carbamate

[0320] 2-(tert-Butoxycarbonylamino)acetic acid 16a (31.54 mg, 180.07 μmol, commercially available), N,N-diisopropylethylamine (62.06 mg, 480.18 μmol), and O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (68.47 mg, 180.07 μmol) were dissolved in N,N-dimethylformamide (2 mL). After stirring for 5 min, (S)-1-(1-((5-(4-((7-azaspiro[3.5]non-2-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol 1 (50.00 mg, 120.04 μmol) was added and the reaction was allowed to proceed at room temperature overnight. After the reaction was completed, water (50 mL) was added and extracted with ethyl acetate (50 mL×3). The organic phases were combined, washed with saturated sodium chloride (20 mL), and dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product, which was then separated by preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H2O, mobile phase B: CH3CN) to give (S)-tert-butyl(2-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]non-7-yl)-2-oxoethyl)carbamate 16b (20 mg) in a yield of 29.04%.

[0321] MS m / z(ESI):574.3[M+1].

[0322] Step 2

[0323] (S)-2-amino-1-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)ethan-1-one

[0324] (S)-2-Amino-1-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)ethan-1-one

[0325] Tert-butyl (S)-(2-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)-2-oxoethyl)carbamate 16b (20 mg, 34.86 μmol) was dissolved in dichloromethane (2 mL), and trifluoroacetic acid (0.3 ml, 2.63 mmol) was added dropwise, and the reaction was carried out at room temperature for 1.5 hours. After the reaction was completed, the mixture was concentrated and subjected to preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H2O, mobile phase B: CH3CN) to give (S)-2-amino-1-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)ethan-1-one 16 (3 mg) in a yield of 3.91%.

[0326] MS m / z(ESI):474.0[M+1].

[0327] Example 17

[0328] (S)-2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonane-7-carbaldehyde

[0329] (S)-2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonane-7-carbaldehyde

[0330] (S)-1-(1-((5-(4-((7-azaspiro[3.5]non-2-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol 1 (30 mg, 56.55 μmol) was dissolved in formamide (2 mL), and triethylamine (28.61 mg, 282.73 μmol) was added dropwise, and the reaction was carried out at room temperature for 6 hours. After the reaction was completed, the mixture was concentrated and subjected to preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H2O, mobile phase B: CH3CN) to give (S)-2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonane-7-carbaldehyde 17 (3 mg) in a yield of 9.02%.

[0331] MS m / z(ESI):445.2[M+1].

[0332] Example 18

[0333] (S)-2-amino-3-hydroxy-1-(2-((4-(3-((2-((S)-1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)propan-1-one

[0334] (S)-2-Amino-3-hydroxy-1-(2-((4-(3-((2-((S)-1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)propan-1-one

[0335] first step

[0336] tert-butyl

[0337] ((S)-3-hydroxy-1-(2-((4-(3-((2-((S)-1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phe nyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)-1-oxopropan-2-yl)carbamate

[0338] tert-Butyl ((S)-3-hydroxy-1-(2-((4-(3-((2-((S)-1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)-1-oxopropan-2-yl)carbamate

[0339] (tert-Butoxycarbonyl)-L-serine 18a (29.01 mg, 141.37 μmol, commercially available), N,N-diisopropylethylamine (60.90 mg, 471.22 μmol), and O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (53.75 mg, 141.37 μmol) were dissolved in N,N-dimethylformamide (2 mL). After stirring for 5 minutes, (S)-1-(1-((5-(4-((7-azaspiro[3.5]non-2-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol 1 (50 mg, 94.24 μmol) was added and the reaction was allowed to proceed at room temperature overnight. After completion of the reaction, water (50 mL) was added to quench the reaction, and the mixture was extracted three times with ethyl acetate (50 ml×3). The organic phases were combined, washed with saturated aqueous sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: System B) to give tert-butyl ((S)-3-hydroxy-1-(2-((4-(3-((2-((S)-1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]non-7-yl)-1-oxopropan-2-yl)carbamate 18b (20 mg) in a yield of 35.15%.

[0340] MS m / z(ESI):604.3[M+1].

[0341] Step 2

[0342] (S)-2-amino-3-hydroxy-1-(2-((4-(3-((2-((S)-1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)propan-1-one

[0343] (S)-2-Amino-3-hydroxy-1-(2-((4-(3-((2-((S)-1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)propan-1-one

[0344] Tert-butyl ((S)-3-hydroxy-1-(2-((4-(3-((2-((S)-1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]non-7-yl)-1-oxopropan-2-yl)carbamate 18b (20 mg, 33.13 μmol) was dissolved in dichloromethane (3 mL), and trifluoroacetic acid (0.5 g, 4.39 mmol) was added dropwise, and the reaction was carried out at room temperature for 3 hours. After the reaction was completed, the mixture was concentrated and subjected to preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H2O, mobile phase B: CH3CN) to give (S)-2-amino-3-hydroxy-1-(2-((4-(3-((2-((S)-1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)propan-1-one 18 (5 mg) in a yield of 24.44%.

[0345] MS m / z(ESI):504.0[M+1].

[0346] Example 19

[0347] (S)-2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonane-7-carboxamide

[0348] (S)-2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonane-7-carboxamide

[0349] (S)-1-(1-((5-(4-((7-azaspiro[3.5]non-2-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol 1 (50 mg, 120.04 μmol), tetrahydrofuran (2 mL), and triethylamine (36.44 mg, 360.13 μmol) were added to the reaction flask in sequence, and finally trimethylsilyl isocyanate 19a (17.98 mg, 156.06 μmol, commercially available) was added and the reaction was allowed to react at room temperature overnight. After the reaction was completed, the mixture was concentrated and subjected to preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H2O, mobile phase B: CH3CN) to give (S)-2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonane-7-carboxamide 19 (10 mg) in a yield of 13.80%.

[0350] MS(ESI):460.2[M+1].

[0351] Example 20

[0352] (S)-2-(6-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-2-azaspiro[3.3]heptan-2-yl)acetamide

[0353] (S)-2-(6-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-2-azaspiro[3.3]hept-2-yl)acetamide

[0354] 2-Bromoacetamide 13a (30 mg, 217.45 μmol, commercially available), (S)-1-(1-((5-(4-((2-azaspiro[3.3]hept-6-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol 6 (84.47 mg, 217.45 μmol), potassium carbonate (90.16 mg, 652.35 μmol), and sodium iodide (32.62 mg, 217.45 μmol) were added sequentially to N,N-dimethylformamide (1.5 mL) and reacted at 80°C for 4 hours. After the reaction was substantially completed, water (50 mL) was added to quench the reaction, and the mixture was extracted three times with ethyl acetate (50 mL×3). The organic phases were combined and concentrated, and subjected to preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H2O, mobile phase B: CH3CN) to give (S)-2-(6-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-2-azaspiro[3.3]hept-2-yl)acetamide 20 (3 mg) in a yield of 2.34%.

[0355] MS m / z(ESI):446.1[M+1].

[0356] Example 21

[0357] (S)-6-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-2-azaspiro[3.3]heptane-2-carboxamide

[0358] (S)-6-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-2-azaspiro[3.3]heptane-2-carboxamide

[0359] (S)-1-(1-((5-(4-((2-Azaspiro[3.3]hept-6-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol 6 (35 mg, 90.10 μmol), tetrahydrofuran (2 mL), and triethylamine (27.35 mg, 270.30 μmol) were added to the reaction flask in sequence, and trimethylsilyl isocyanate 19a (13.49 mg, 117.13 μmol, commercially available) was added, and the reaction was allowed to react at room temperature overnight. After the reaction was completed, the mixture was concentrated and subjected to preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H2O, mobile phase B: CH3CN) to give (S)-6-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-2-azaspiro[3.3]heptane-2-carboxamide 21 (9 mg) with a yield of 17.40%.

[0360] MS m / z(ESI):432.0[M+1].

[0361] Example 22

[0362] (S)-6-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-2-azaspiro[3.3]heptane-2-carbaldehyde

[0363] (S)-6-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-2-azaspiro[3.3]heptane-2-carbaldehyde

[0364] (S)-1-(1-((5-(4-((2-Azaspiro[3.3]hept-6-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol 6 (35 mg, 90.10 μmol), formamide (2 mL), and triethylamine (27.35 mg, 270.30 μmol) were added sequentially into a single-necked flask and reacted at room temperature overnight. After the reaction was completed, the mixture was concentrated and subjected to preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H2O, mobile phase B: CH3CN) to give (S)-6-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-2-azaspiro[3.3]heptane-2-carbaldehyde 22 (5 mg) in a yield of 9.94%.

[0365] MS m / z(ESI):417.2[M+1].

[0366] Example 23

[0367] (S)-2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-6-azaspiro[3.4]octane-6-carbaldehyde

[0368] (S)-2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-6-azaspiro[3.4]octane-6-carbaldehyde

[0369] (S)-1-(1-((5-(4-((6-Azaspiro[3.4]octan-2-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol 3 (10 mg, 24.85 μmol) was dissolved in formamide (2 mL), triethylamine (25.14 mg, 248.45 μmol) was added, and the reaction was allowed to react at room temperature overnight. After the reaction was completed, the mixture was concentrated and subjected to preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H2O, mobile phase B: CH3CN) to give (S)-2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-6-azaspiro[3.4]octane-6-carbaldehyde 23 (2.40 mg) in a yield of 15.08%.

[0370] MS m / z(ESI):431.3[M+1].

[0371] Example 24

[0372] (S)-2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-6-azaspiro[3.4]octane-6-carboxamide

[0373] (S)-2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-6-azaspiro[3.4]octane-6-carboxamide

[0374] (S)-1-(1-((5-(4-((6-Azaspiro[3.4]octan-2-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol 3 (36.26 mg, 90.10 μmol) was dissolved in tetrahydrofuran (2 mL), and triethylamine (27.35 mg, 270.30 μmol) and trimethylsilyl isocyanate 19a (13.49 mg, 117.13 μmol) were added, and the reaction was carried out at room temperature for 4 hours. After completion of the reaction, the mixture was concentrated and subjected to preparative liquid separation (AKZONOBEL Kromasil column; 250 × 21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA + H₂O, mobile phase B: CH₃CN) to afford (S)-2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-6-azaspiro[3.4]octane-6-carboxamide 24 (3 mg) in a 5.36% yield. MS m / z (ESI): 446.0 [M+1].

[0375] Example 25

[0376] (S)-1-(1-((5-(4-((7-((1H-pyrazol-5-yl)methyl)-7-azaspiro[3.5]nonan-2-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol

[0377] (S)-1-(1-((5-(4-((7-((1H-pyrazol-5-yl)methyl)-7-azaspiro[3.5]non-2-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol

[0378] first step

[0379] tert-butyl

[0380] (S)-5-((2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)methyl)-1H-pyrazole-1-carboxylate

[0381] (S)-tert-Butyl 5-((2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)methyl)-1H-pyrazole-1-carboxylate

[0382] (S)-1-(1-((5-(4-((7-azaspiro[3.5]non-2-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol 1 (50 mg, 120.04 μmol), tert-butyl 3-(bromomethyl)pyrazole-1-carboxylate 25a (31.35 mg, 120.04 μmol, commercially available), and potassium carbonate (19.91 mg, 144.05 μmol) were added sequentially to N,N-dimethylformamide (1.5 mL) and reacted at 75 °C for 8 h. After the reaction was completed, water (50 mL) was added, and ethyl acetate (50 mL×3) was added and extracted three times. The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to give the crude product, which was then purified by silica gel column chromatography (eluent: System B) to give the product (S)-5-((2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]non-7-yl)methyl)-1H-pyrazole-1-carboxylic acid tert-butyl ester 25b (40 mg) in a yield of 55.84%.

[0383] MS m / z(ESI):497.3[M-100+1].

[0384] Step 2

[0385] (S)-1-(1-((5-(4-((7-((1H-pyrazol-5-yl)methyl)-7-azaspiro[3.5]nonan-2-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol

[0386] (S)-1-(1-((5-(4-((7-((1H-pyrazol-5-yl)methyl)-7-azaspiro[3.5]non-2-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol

[0387] (S)-tert-Butyl 5-((2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]non-7-yl)methyl)-1H-pyrazole-1-carboxylate 25b (40 mg, 67.03 μmol) was dissolved in dichloromethane (1 mL), and trifluoroacetic acid (67.03 μmol, 0.25 mL) was slowly added. The reaction was completed after 3 hours at room temperature. The product was concentrated and subjected to preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H2O, mobile phase B: CH3CN) to give (S)-1-(1-((5-(4-((7-((1H-pyrazol-5-yl)methyl)-7-azaspiro[3.5]non-2-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol 25 (10 mg) in a yield of 23.21%.

[0388] MS m / z(ESI):497.3[M+1].

[0389] Example 26

[0390] (S)-2-hydroxy-1-(6-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-2-azaspiro[3.3]heptan-2-yl)ethan-1-one

[0391] (S)-2-Hydroxy-1-(6-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-2-azaspiro[3.3]hept-2-yl)ethan-1-one

[0392] 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (25.91 mg, 135.15 μmol), 1-hydroxybenzotriazole (18.26 mg, 135.15 μmol), N-methylmorpholine (20.05 mg, 198.22 μmol), and 2-hydroxyacetic acid 12a (10.28 mg, 135.15 μmol) were dissolved in dichloromethane (2 mL). After 5 minutes of reaction, (S)-1-(1-((5-(4-((2-azaspiro[3.3]hept-6-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol 6 (35 mg, 90.10 μmol) was added and the reaction was allowed to proceed at room temperature overnight. After the reaction was completed, water (50 mL) was added, and dichloromethane (50 mL×3) was added and extracted three times. The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to obtain a crude product. The crude product was separated by preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H2O, mobile phase B: CH3CN) to obtain (S)-2-hydroxy-1-(6-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-2-azaspiro[3.3]hept-2-yl)ethan-1-one 26 (2.80 mg) in a yield of 5.27%.

[0393] MS m / z(ESI):447.2[M+1].

[0394] Example 27

[0395] (S)-1-(1-((5-(4-((7-((1H-imidazol-2-yl)methyl)-7-azaspiro[3.5]nonan-2-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol

[0396] (S)-1-(1-((5-(4-((7-((1H-imidazol-2-yl)methyl)-7-azaspiro[3.5]non-2-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol

[0397] (S)-1-(1-((5-(4-((7-Azaspiro[3.5]non-2-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol 1 (50 mg, 120.04 μmol), 2-(bromomethyl)-1H-imidazole 27a (26.14 mg, 108.04 μmol), sodium iodide (1.99 mg, 12.00 μmol), and potassium carbonate (36.50 mg, 264.10 μmol) were added sequentially to N,N-dimethylformamide (1.5 mL) and heated to 65°C for 8 hours. After the reaction was substantially completed, the reaction was filtered and the filtrate was subjected to preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H2O, mobile phase B: CH3CN) to give (S)-1-(1-((5-(4-((7-((1H-imidazol-2-yl)methyl)-7-azaspiro[3.5]non-2-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol 27 (10 mg) in a yield of 12.96%.

[0398] MS(ESI):497.1[M+1].

[0399] Example 28

[0400] (S)-2-hydroxy-1-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-6-azaspiro[3.4]octan-6-yl)ethan-1-one

[0401] (S)-2-Hydroxy-1-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-6-azaspiro[3.4]octan-6-yl)ethan-1-one

[0402] 2-Hydroxyacetic acid 12a (14.17 mg, 186.34 μmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (35.72 mg, 186.34 μmol), 1-hydroxybenzotriazole (25.18 mg, 186.34 μmol), and N-methylmorpholine (27.64 mg, 273.30 μmol) were added to a dichloromethane (2 mL) solution and reacted for 5 minutes. Then, (S)-1-(1-((5-(4-((6-azaspiro[3.4]octan-2-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol 3 (50 mg, 124.23 μmol) was added and reacted at room temperature for 16 hours. After the reaction was completed, water (50 mL) was added, and the mixture was extracted three times with dichloromethane (50 mL×3). The organic layers were combined and concentrated, and subjected to preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H2O, mobile phase B: CH3CN) to obtain (S)-2-hydroxy-1-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-6-azaspiro[3.4]octan-6-yl)ethan-1-one 28 (5 mg) in a yield of 6.30%.

[0403] MS m / z(ESI):461.2[M+1].

[0404] Example 29

[0405] (S)-3-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)-3-oxopropanenitrile

[0406] (S)-3-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)-3-oxopropionitrile

[0407] 2-Cyanoacetic acid 29a (15.32 mg, 180.07 μmol, commercially available), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (34.52 mg, 180.07 μmol), 1-hydroxybenzotriazole (24.33 mg, 180.07 μmol), and N-methylmorpholine (26.71 mg, 264.10 μmol) were added to a solution of N,N-dimethylformamide (1.5 mL). After reacting for 5 minutes, (S)-1-(1-((5-(4-((7-azaspiro[3.5]non-2-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol 1 (50 mg, 120.04 μmol) was added and reacted at room temperature for 16 hours. After the reaction was completed, water (50 mL) was added, and the mixture was extracted three times with dichloromethane (50 mL×3). The organic phases were combined, concentrated, and subjected to preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H2O, mobile phase B: CH3CN) to obtain (S)-3-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]non-7-yl)-3-oxopropionitrile 29 (3 mg) in a yield of 3.97%.

[0408] MS m / z(ESI):484.2[M+1].

[0409] Example 30

[0410] (S)-1-(1-((5-(4-((7-(azetidin-3-yl)-7-azaspiro[3.5]nonan-2-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol

[0411] (S)-1-(1-((5-(4-((7-(azetidin-3-yl)-7-azaspiro[3.5]non-2-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol

[0412] first step

[0413] tert-butyl

[0414] (S)-3-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)azetidine-1-carboxylate

[0415] (S)-tert-Butyl 3-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]nonan-7-yl)azetidine-1-carboxylate

[0416] (S)-1-(1-((5-(4-((7-azaspiro[3.5]non-2-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol 1 (26.81 mg, 64.36 μmol) and tert-butyl 3-oxoazetidine-1-carboxylate 30a (16.53 mg, 96.53 μmol, commercially available) were dissolved in a mixed solution of 1,2-dichloroethane (0.5 mL) and methanol (0.5 mL) at room temperature. Acetic acid (20 mg, 64.36 μmol) was added dropwise, and the mixture was reacted at 50°C for 6 hours. Then, sodium cyanoborohydride (12.13 mg, 193.07 μmol) was slowly added at room temperature, and the reaction was carried out at room temperature for 16 hours. After completion of the reaction, water (1 mL) was added to quench the reaction, and the mixture was concentrated. Preparative liquid separation (AKZONOBEL Kromasil column; 250 × 21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA + H₂O, mobile phase B: CH₃CN) afforded (S)-tert-butyl 3-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]non-7-yl)azetidine-1-carboxylate 30b (25 mg) in a 67.95% yield. MS m / z (ESI): 572.4 [M+1].

[0417] Step 2

[0418] (S)-1-(1-((5-(4-((7-(azetidin-3-yl)-7-azaspiro[3.5]nonan-2-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol

[0419] (S)-1-(1-((5-(4-((7-(azetidin-3-yl)-7-azaspiro[3.5]non-2-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol

[0420] At room temperature, tert-butyl (S)-3-(2-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-7-azaspiro[3.5]non-7-yl)azetidine-1-carboxylate 30b (25 mg, 43.73 μmol) was dissolved in dichloromethane (2 mL), and trifluoroacetic acid (0.5 g, 4.39 mmol) was added dropwise, and the reaction was carried out at room temperature for 4 hours. After completion of the reaction, the mixture was concentrated and subjected to preparative liquid separation (AKZONOBEL Kromasil column; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H₂O, mobile phase B: CH₃CN) to afford (S)-1-(1-((5-(4-((7-(azetidin-3-yl)-7-azaspiro[3.5]non-2-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol 30 (9 mg) in a yield of 31.63%. MS m / z (ESI): 472.3 [M+1].

[0421] Example 31

[0422] (S)-2-(6-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-2-azaspiro[3.3]heptan-2-yl)ethan-1-ol

[0423] (S)-2-(6-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-2-azaspiro[3.3]hept-2-yl)ethan-1-ol

[0424] At room temperature, (S)-1-(1-((5-(4-((2-azaspiro[3.3]hept-6-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol 6 (25 mg, 49.75 μmol), 2-bromoethanol 11a (6.84 mg, 54.73 μmol, commercially available), and potassium carbonate (27.51 mg, 199.01 μmol) were added to acetonitrile (0.5 mL) and reacted at 60 °C for 12 h. After the reaction was completed, the product was filtered and the filtrate was concentrated. The product was separated by preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H2O, mobile phase B: CH3CN) to give (S)-2-(6-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-2-azaspiro[3.3]hept-2-yl)ethan-1-ol (8 mg) in a yield of 26.48%.

[0425] MS m / z(ESI):433.0[M+1].

[0426] Example 32

[0427] (S)-3-(6-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-2-azaspiro[3.3]heptan-2-yl)-3-oxopropanenitrile

[0428] (S)-3-(6-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-2-azaspiro[3.3]hept-2-yl)-3-oxopropionitrile

[0429] At room temperature, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (57.23 mg, 298.52 μmol), 1-hydroxybenzotriazole (40.34 mg, 298.52 μmol), N-methylmorpholine (84.55 mg, 835.85 μmol), and 2-cyanoacetic acid 32a (30.47 mg, 358.22 μmol, commercially available) were dissolved in dichloromethane (2 mL). After reacting for 5 minutes, (S)-1-(1-((5-(4-((2-azaspiro[3.3]hept-6-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol 6 (60 mg, 119.41 μmol) was added and the reaction was allowed to proceed overnight at room temperature. After the reaction was completed, water (50 mL) was added, and dichloromethane (50 mL×3) was added for extraction. The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to give a crude product. The crude product was subjected to preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H2O, mobile phase B: CH3CN) to give (S)-3-(6-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-2-azaspiro[3.3]hept-2-yl)-3-oxopropionitrile 32 (20 mg) in a yield of 27.94%.

[0430] MS m / z(ESI):456.2[M+1].

[0431] Example 33

[0432] (S)-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(pyridin-4-yl)methanone

[0433] (S)-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(pyridin-4-yl)methanone

[0434] first step

[0435] tert-butyl 4-((trimethylsilyl)ethynyl)-3,6-dihydropyridine-1(2H)-carboxylate

[0436] tert-Butyl 4-((Trimethylsilyl)ethynyl)-3,6-dihydropyridine-1(2H)-carboxylate

[0437] Tert-butyl 4-(((trifluoromethyl)sulfonyl)oxy)-3,6-dihydropyridine-1(2H)-carboxylate 33a (1 g, 3.02 mmol, commercially available), ethynyltrimethylsilane 33b (444.68 mg, 4.53 mmol, 639.83 μL, commercially available), bis(triphenylphosphine)palladium(II) dichloride (352.28 mg, 452.75 μmol), triethylamine (305.43 mg, 3.02 mmol), cuprous iodide (57.48 mg, 301 .83 μmol) were sequentially added to a toluene (3 mL) solution, and the mixture was stirred at room temperature for 16 h. The reaction solution was distilled under reduced pressure to remove the organic solvent, and water (20 mL) was added. The mixture was extracted with ethyl acetate (20 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. The residue was purified by silica gel column chromatography (eluent: System A) to obtain tert-butyl 4-((trimethylsilyl)ethynyl)-3,6-dihydropyridine-1(2H)-carboxylate 33c (820 mg) in a yield of 97.22%.

[0438] MS m / z(ESI):224.1[M+1-56].

[0439] Step 2

[0440] tert-butyl 4-ethynyl-3,6-dihydropyridine-1(2H)-carboxylate

[0441] tert-Butyl 4-ethynyl-3,6-dihydropyridine-1(2H)-carboxylate

[0442] tert-Butyl 4-((trimethylsilyl)ethynyl)-3,6-dihydropyridine-1(2H)-carboxylate 33c (800 mg, 2.86 mmol) was added to methanol (2 mL), followed by potassium fluoride (332.65 mg, 5.73 mmol), and stirred at room temperature for 12 hours. The reaction mixture was distilled under reduced pressure to remove the organic solvent, and water (20 mL) was added. The mixture was extracted with ethyl acetate (20 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. The residue was purified by silica gel column chromatography (eluent: System A) to obtain tert-butyl 4-ethynyl-3,6-dihydropyridine-1(2H)-carboxylate 33d (370 mg) in a yield of 62.36%.

[0443] MS m / z(ESI):152.1[M+1-56].

[0444] Step 3

[0445] tert-butyl

[0446] 4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridine-1(2H)-carboxylate

[0447] tert-Butyl 4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridine-1(2H)-carboxylate

[0448] 5-(4-iodophenyl)-3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazole 1i (800 mg, 1.67 mmol), tert-butyl 4-ethynyl-3,6-dihydropyridine-1(2H)-carboxylate 33d (363.24 mg, 1.75 mmol), allylpalladium(II) chloride dimer (121.84 mg, 333.81 μmol), triethylenediamine (374.44 mg, 3.34 mmol), and tri-tert-butylphosphine (10% in toluene solution) (67.54 mg, 333.81 μmol) were added sequentially to acetonitrile (2 mL) and reacted at room temperature overnight. After completion of the reaction, water (30 mL) was added to quench the reaction, and the mixture was extracted with ethyl acetate (100 mL × 3). The combined organic phases were washed with saturated brine (100 mL) and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (eluent: System B) to give tert-butyl 4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridine-1(2H)-carboxylate 33e (660 mg) in a yield of 70.78%. MS m / z (ESI): 559.2 [M+1].

[0449] Step 4

[0450] (S)-1-(1-((5-(4-((1,2,3,6-tetrahydropyridin-4-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol

[0451] (S)-1-(1-((5-(4-((1,2,3,6-tetrahydropyridin-4-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol

[0452] Tert-butyl 4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridine-1(2H)-carboxylate 33e (660 mg, 1.18 mmol) was added to dichloromethane (1.5 mL), followed by the dropwise addition of trifluoroacetic acid (0.5 mL), and stirring was continued at room temperature for 1 hour. After the reaction was complete, saturated NaHCO3 solution was added under ice bath until no bubbles were generated (pH>7), and the mixture was extracted with ethyl acetate (20 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate, and distilled under reduced pressure. The residue was purified by silica gel column chromatography (eluent: System B) to give (S)-1-(1-((5-(4-((1,2,3,6-tetrahydropyridin-4-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol 33f (320 mg) in a yield of 72.34%.

[0453] MS m / z(ESI):375.2[M+1].

[0454] Step 5

[0455] (S)-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(pyridin-4-yl)methanone

[0456] (S)-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(pyridin-4-yl)methanone

[0457] Isonicotinic acid 33g (32.88 mg, 267.07 μmol, commercially available), (S)-1-(1-((5-(4-((1,2,3,6-tetrahydropyridin-4-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol 33f (50 mg, 133.53 μmol), N,N-diisopropylethylamine (34.52 mg, 267.07 μmol), and (7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (50.77 mg, 133.53 μmol) were added sequentially to N,N-dimethylformamide (1 mL) and stirred at room temperature for 16 hours. After the reaction was completed, water (50 mL) was added, and dichloromethane (50 mL×3) was added for extraction. The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to obtain a crude product. The crude product was subjected to preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H2O, mobile phase B: CH3CN) to obtain (S)-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(pyridin-4-yl)methanone 33 (13.2 mg) in a yield of 15.99%.

[0458] MS m / z(ESI):480.3[M+1].

[0459] Example 34

[0460] (S)-4-((4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3, 6-dihydropyridin-1(2H)-yl)methyl)benzoic acid

[0461] (S)-4-((4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methyl)benzoic acid

[0462] 4-Formylbenzoic acid 34a (24.06 mg, 160.24 μmol, commercially available), (S)-1-(1-((5-(4-((1,2,3,6-tetrahydropyridin-4-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol 33f (50 mg, 133.53 μmol), and acetic acid (8.02 mg, 133.53 μmol) were added sequentially to dichloromethane (1 mL) and stirred at room temperature for 1 hour. Sodium triacetoxyborohydride (28.30 mg, 133.53 μmol) was then added and stirred at room temperature for 16 hours. After the reaction was completed, water (50 mL) was added, and dichloromethane (50 mL×3) was added for extraction. The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to give a crude product. The crude product was subjected to preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H2O, mobile phase B: CH3CN) to give (S)-4-((4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methyl)benzoic acid 34 (7.35 mg) in a yield of 8.66%.

[0463] MS m / z(ESI):509.0[M+1].

[0464] Example 35

[0465] ((1R,5S)-bicyclo[3.1.0]hexan-6-yl)(4-((4-(3-((2-((S)-1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0466] ((1R,5S)-Bicyclo[3.1.0]hexan-6-yl)(4-((4-(3-((2-((S)-1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0467] first step

[0468] 3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)-5-(4-((trimethylsilyl)ethynyl)phenyl)isoxazole

[0469] 3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)-5-(4-((trimethylsilyl)ethynyl)phenyl)isoxazole

[0470] 5-(4-iodophenyl)-3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazole 1i (8 g, 16.69 mmol), ethynyltrimethylsilane 33b (3.28 g, 33.38 mmol, 4.72 mL, commercially available), allylpalladium(II) chloride dimer (609.21 mg, 1.67 mmol), triethylenediamine (3.74 g, 33.38 mmol), and tri-tert-butylphosphine (10% toluene solution) (3.38 g, 1.67 mmol) were added to acetonitrile (20 mL) in sequence, and the argon atmosphere was replaced three times. The mixture was stirred at room temperature for 12 hours. The system was concentrated to dryness under reduced pressure, water (50 mL) was added, and extraction was carried out with ethyl acetate (50 mL×3). The combined organic phases were dried over anhydrous sodium sulfate and then distilled under reduced pressure. The residue was purified by silica gel column chromatography (eluent: System A) to give 3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)-5-(4-((trimethylsilyl)ethynyl)phenyl)isoxazole 35a (7.5 g) in a yield of 99.94%.

[0471] MS m / z(ESI):450.3[M+1].

[0472] Step 2

[0473] 5-(4-ethynylphenyl)-3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazole

[0474] 5-(4-ethynylphenyl)-3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazole

[0475] 3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)-5-(4-((trimethylsilyl)ethynyl)phenyl)isoxazole 35a (7.5 g, 16.68 mmol) and potassium fluoride (1.94 g, 33.36 mmol) were added to methanol (50 mL) in sequence. The reaction was completed after continuous stirring at room temperature for 5 hours. Water was added to obtain the mixture. The product was stirred for 2 hours at room temperature for 1 h (50 mL), extracted with ethyl acetate (50 mL × 3), and the combined organic phases were dried over anhydrous sodium sulfate and then distilled under reduced pressure. The residue was purified by silica gel column chromatography (eluent: System A) to give 5-(4-ethynylphenyl)-3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazole 35b (5.8 g) in a yield of 92.21%.

[0476] MS m / z(ESI):378.2[M+1].

[0477] Step 3

[0478] 1,2,3,6-tetrahydropyridin-4-yl trifluoromethanesulfonate

[0479] 1,2,3,6-Tetrahydropyridin-4-yl trifluoromethanesulfonate

[0480] Tert-butyl 4-(((trifluoromethyl)sulfonyl)oxy)-3,6-dihydropyridine-1(2H)-carboxylate 33a (1.7 g, 5.13 mmol) was added to dichloromethane (1.5 mL), followed by the dropwise addition of trifluoroacetic acid (0.5 mL). The mixture was stirred at room temperature for 1 hour. After the reaction was complete, saturated NaHCO3 solution was added to the reaction solution until no bubbles were generated and the pH was > 7. Water (50 mL) was then added, and dichloromethane (50 mL × 3) was added for extraction. The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to give 1,2,3,6-tetrahydropyridin-4-yl trifluoromethanesulfonate 35c (1.1 g) in a yield of 92%. The crude product was used directly in the next reaction.

[0481] MS m / z(ESI):232.1[M+1].

[0482] Step 4

[0483] 1-(2,2,2-trifluoroacetyl)-1,2,3,6-tetrahydropyridin-4-yl trifluoromethanesulfonate

[0484] 1-(2,2,2-Trifluoroacetyl)-1,2,3,6-tetrahydropyridin-4-yl trifluoromethanesulfonate

[0485] 1,2,3,6-Tetrahydropyridin-4-yl trifluoromethanesulfonate 35c (900 mg, 3.89 mmol) and triethylamine (393.92 mg, 3.89 mmol) were added sequentially to dichloromethane (10 mL) under an ice bath. Trifluoroacetic anhydride (820 mg, 3.89 mmol) was then added dropwise to the mixed solution. The mixture was then gradually warmed to room temperature and stirred for 20 minutes. After the reaction was complete, water (50 mL) was added and the mixture was extracted with ethyl acetate (50 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate and evaporated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: System A) to give 1-(2,2,2-trifluoroacetyl)-1,2,3,6-tetrahydropyridin-4-yl trifluoromethanesulfonate 35d (1.17 g) in a yield of 91%.

[0486] MS m / z(ESI):328.0[M+1].

[0487] Step 5

[0488] 2,2,2-trifluoro-1-(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)ethan-1-one

[0489] 2,2,2-Trifluoro-1-(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)ethan-1-one

[0490] Under argon atmosphere, 1-(2,2,2-trifluoroacetyl)-1,2,3,6-tetrahydropyridin-4-yl trifluoromethanesulfonate 35d (1.13 g, 3.44 mmol), 5-(4-ethynylphenyl)-3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazole 35b (1 g, 2.65 mmol), cuprous iodide (50.46 mg, 264.95 μmol), bis(triphenylphosphine)palladium(II) dichloride (412.30 mg, 529.89 μmol), and triethylamine (268.10 mg, 2.65 mmol) were added sequentially to toluene (5 mL) and stirred at room temperature for 16 hours. After the reaction was completed, the reaction solution was distilled under reduced pressure to remove the organic solvent. The residue was purified by silica gel column chromatography (eluent: System A) to give 2,2,2-trifluoro-1-(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)ethan-1-one 35e (400 mg) in a yield of 27.22%.

[0491] MS m / z(ESI):555.3[M+1].

[0492] Step 6

[0493] 3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)-5-(4-((1,2,3,6-tetrahydropyridin-4-yl)ethynyl)phenyl)isoxazole

[0494] 3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)-5-(4-((1,2,3,6-tetrahydropyridin-4-yl)ethynyl)phenyl)isoxazole

[0495] 2,2,2-Trifluoro-1-(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)ethan-1-one 35e (400 mg, 721.29 μmol) and potassium carbonate (797.52 mg, 5.77 mmol) were added sequentially to a mixture of methanol (10 mL) and water (0.5 mL). The solution was replaced with argon three times, and the reaction was stirred at 70°C for 2 hours. After the reaction was complete, the reaction solution was distilled under reduced pressure to remove the organic solvent. The crude product was purified by silica gel column chromatography (eluent: System B) to give 3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)-5-(4-((1,2,3,6-tetrahydropyridin-4-yl)ethynyl)phenyl)isoxazole 35f (180 mg) in a yield of 54.42%.

[0496] MS m / z(ESI):459.3[M+1].

[0497] Step 7

[0498] bicyclo[3.1.0]hexan-6-yl(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0499] Bicyclo[3.1.0]hexane-6-carboxylic acid 35 g (16.51 mg, 130.85 μmol, commercially available), 3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)-5-(4-((1,2,3,6-tetrahydropyridin-4-yl)ethynyl)phenyl)isoxazole 35f (30 mg, 65.42 μmol), N-methylmorpholine (19.85 mg, 196.27 μmol), 1-hydroxybenzotriazole (17.68 mg, 130.85 μmol), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (25.08 mg, 130.85 μmol) were added to dichloromethane (1 mL) in sequence, and stirring was continued at room temperature for 16 hours. After the reaction was complete, water (50 mL) was added, and dichloromethane (50 mL×3) was added for extraction. The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to give bicyclo[3.1.0]hexan-6-yl(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone 35h (37 mg), and the crude product was used directly in the next reaction.

[0500] MS m / z(ESI):567.2[M+1].

[0501] Step 8

[0502] ((1R,5S)-bicyclo[3.1.0]hexan-6-yl)(4-((4-(3-((2-((S)-1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0503] ((1R,5S)-Bicyclo[3.1.0]hexan-6-yl)(4-((4-(3-((2-((S)-1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0504] Bicyclo[3.1.0]hexan-6-yl(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone 35h (37 mg, 65.29 μmol) was added to dichloromethane (1.5 mL), followed by the dropwise addition of trifluoroacetic acid (0.5 mL), and stirring was continued for half an hour at room temperature. The crude product was concentrated and subjected to preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H2O, mobile phase B: CH3CN) to give ((1R,5S)-bicyclo[3.1.0]hexan-6-yl)(4-((4-(3-((2-((S)-1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone 35 (4.05 mg) in a yield of 9.88%.

[0505] MS m / z(ESI):483.2[M+1].

[0506] Example 36

[0507] (S)-cyclopropyl(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0508] (S)-Cyclopropyl(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0509] first step

[0510] cyclopropyl(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0511] Cyclopropyl(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0512] Cyclopropanecarboxylic acid 36a (11.26 mg, 130.85 μmol), 3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)-5-(4-((1,2,3,6-tetrahydropyridin-4-yl)ethynyl)phenyl)isoxazole 35f (30 mg, 65.42 μmol), N-methylmorpholine (19.85 mg, 196.27 μmol), 1-hydroxybenzotriazole (17.68 mg, 130.85 μmol), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (25.08 mg, 130.85 μmol) were added sequentially to dichloromethane (1 mL) and stirred at room temperature for 6 hours. After the reaction was complete, water (50 mL) was added, followed by extraction with dichloromethane (50 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to afford cyclopropyl(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone 36b (34 mg) in a 98.68% yield. The crude product was used directly in the next reaction.

[0513] MS m / z(ESI):527.3[M+1].

[0514] Step 2

[0515] (S)-cyclopropyl(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0516] (S)-Cyclopropyl(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0517] Cyclopropyl(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone 36b (34 mg, 64.56 μmol) was added to dichloromethane (1.5 mL), followed by the dropwise addition of trifluoroacetic acid (0.5 mL), and stirring was continued for half an hour at room temperature. The crude product was concentrated and subjected to preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H2O, mobile phase B: CH3CN) to give (S)-cyclopropyl(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone 36 (25.29 mg) in a yield of 69.68%.

[0518] MS m / z(ESI):443.2[M+1].

[0519] Example 37

[0520] (S)-1-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)-2-(1H-1,2,4-triazol-1-yl)ethan-1-one

[0521] (S)-1-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)-2-(1H-1,2,4-triazol-1-yl)ethan-1-one

[0522] first step

[0523] 1-(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isox azol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)-2-(1H-1,2,4-triazol-1-yl)ethan-1-one

[0524] 1-(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)-2-(1H-1,2,4-triazol-1-yl)ethan-1-one

[0525] 2-(1H-1,2,4-triazol-1-yl)acetic acid 37a (16.63 mg, 130.85 μmol), 3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)-5-(4-((1,2,3,6-tetrahydropyridin-4-yl)ethynyl)phenyl)isoxazole 35f (30 mg, 65.42 μmol), N-methylmorpholine (19.85 mg, 196.27 μmol), 1-hydroxybenzotriazole (17.68 mg, 130.85 μmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (25 To the reaction mixture were added 1-(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)-2-(1H-1,2,4-triazol-1-yl)ethan-1-one 37b (37 mg, 99.63% yield). The product was used directly in the next reaction.

[0526] MS m / z(ESI):568.4[M+1].

[0527] Step 2

[0528] (S)-1-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)-2-(1H-1,2,4-triazol-1-yl)ethan-1-one

[0529] (S)-1-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)-2-(1H-1,2,4-triazol-1-yl)ethan-1-one

[0530] 1-(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)-2-(1H-1,2,4-triazol-1-yl)ethan-1-one 37b (37 mg, 65.18 μmol) was added to dichloromethane (1.5 mL), followed by the dropwise addition of trifluoroacetic acid (0.5 mL), and stirring was continued for half an hour at room temperature. The crude product was concentrated and subjected to preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H2O, mobile phase B: CH3CN) to give (S)-1-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)-2-(1H-1,2,4-triazol-1-yl)ethan-1-one 37 (27.39 mg) in a yield of 69.62%.

[0531] MS m / z(ESI):484.2[M+1].

[0532] Example 38

[0533] (S)-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(pyrazin-2-yl)methanone

[0534] (S)-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(pyrazin-2-yl)methanone

[0535] first step

[0536] pyrazin-2-yl(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0537] Pyrazin-2-yl(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0538] Pyrazine-2-carboxylic acid 38a (16.24 mg, 130.85 μmol), 3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)-5-(4-((1,2,3,6-tetrahydropyridin-4-yl)ethynyl)phenyl)isoxazole 35f (30 mg, 65.42 μmol), N-methylmorpholine (19.85 mg, 196.27 μmol), 1-hydroxybenzotriazole (17.68 mg, 130.85 μmol), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (25.08 mg, 130.85 μmol) were added sequentially to dichloromethane (1 mL) and stirred at room temperature for 6 hours. After the reaction was complete, water (50 mL) was added, followed by extraction with dichloromethane (50 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to afford pyrazin-2-yl(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone 38b (36 mg) in a 97.45% yield. The crude product was used directly in the next reaction.

[0539] MS m / z(ESI):565.3[M+1].

[0540] Step 2

[0541] (S)-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(pyrazin-2-yl)methanone

[0542] (S)-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(pyrazin-2-yl)methanone

[0543] Pyrazin-2-yl(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone 38b (36 mg, 63.76 μmol) was added to dichloromethane (1.5 mL), followed by the dropwise addition of trifluoroacetic acid (0.5 mL), and stirring was continued for half an hour at room temperature. The crude product was concentrated and subjected to preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H2O, mobile phase B: CH3CN) to give (S)-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(pyrazin-2-yl)methanone 38 (14.76 mg) in a yield of 38.12%.

[0544] MS m / z(ESI):481.2[M+1].

[0545] Example 39

[0546] (S)-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-d ihydropyridin-1(2H)-yl)(pyridazin-4-yl)methanone

[0547] (S)-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(pyridazin-4-yl)methanone

[0548] first step

[0549] pyridazin-4-yl(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0550] Pyridazin-4-yl(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0551] Pyridazine-4-carboxylic acid 39a (16.24 mg, 130.85 μmol, commercially available), 3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)-5-(4-((1,2,3,6-tetrahydropyridin-4-yl)ethynyl)phenyl)isoxazole 35f (30 mg, 65.42 μmol), N-methylmorpholine (19.85 mg, 196.27 μmol), 1-hydroxybenzotriazole (17.68 mg, 130.85 μmol), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (25.08 mg, 130.85 μmol) were added sequentially to dichloromethane (1 mL) and stirred at room temperature for 6 hours. After the reaction was complete, water (50 mL) was added, followed by extraction with dichloromethane (50 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to afford pyridazin-4-yl(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone 39b (36 mg) in a 97.45% yield. The crude product was used directly in the next reaction.

[0552] MS m / z(ESI):565.3[M+1].

[0553] Step 2

[0554] (S)-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(pyridazin-4-yl)methanone

[0555] (S)-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(pyridazin-4-yl)methanone

[0556] Pyridazin-4-yl(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone 39b (36 mg, 63.76 μmol) was added to dichloromethane (1.5 mL), followed by the dropwise addition of trifluoroacetic acid (0.5 mL), and stirring was continued for half an hour at room temperature. The crude product was concentrated and subjected to preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H2O, mobile phase B: CH3CN) to give (S)-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(pyridazin-4-yl)methanone 39 (13.22 mg) in a yield of 33.83%.

[0557] MS m / z(ESI):481.2[M+1].

[0558] Example 40

[0559] (S)-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(pyrimidin-5-yl)methanone

[0560] (S)-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(pyrimidin-5-yl)methanone

[0561] first step

[0562] pyrimidin-5-yl(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0563] Pyrimidin-5-yl(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0564] Pyrimidine-5-carboxylic acid 40a (21.65 mg, 174.46 μmol, commercially available), 3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)-5-(4-((1,2,3,6-tetrahydropyridin-4-yl)ethynyl)phenyl)isoxazole 35f (40 mg, 87.23 μmol), 1-hydroxybenzotriazole (23.57 mg, 174.46 μmol), N-methylmorpholine (26.47 mg, 261.69 μmol), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (33.44 mg, 174.46 μmol) were added sequentially to dichloromethane (1.5 mL) and stirred at room temperature for 16 hours. After the reaction was complete, water (50 mL) was added, followed by extraction with dichloromethane (50 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to afford pyrimidin-5-yl(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone 40b (49 mg) in a 99.48% yield. This was used directly in the next reaction.

[0565] MS m / z(ESI):565.3[M+1].

[0566] Step 2

[0567] (S)-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(pyrimidin-5-yl)methanone

[0568] (S)-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(pyrimidin-5-yl)methanone

[0569] Pyrimidin-5-yl(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone 40b (49 mg, 86.78 μmol) was added to dichloromethane (1.5 mL), followed by the dropwise addition of trifluoroacetic acid (0.5 mL), and stirring was continued at room temperature for 1 hour. The crude product was concentrated and subjected to preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H2O, mobile phase B: CH3CN) to give (S)-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(pyrimidin-5-yl)methanone 40 (22.3 mg) in a yield of 42.36%.

[0570] MS m / z(ESI):481.2[M+1]. 1 H NMR (400MHz, DMSO-d6) δ9.30(s,1H),8.94(s,2H),7.88(d,J=8.2Hz,2H),7.79(d,J=2.0Hz,1H),7.72(d,J=2.1Hz,1H),7.62(d,J=8.1Hz,2H ),7.17(s,1H),6.34(s,1H),5.72(s,2H),5.26(q,J=6.6Hz,1H),4.27(s,1H),4.13(s,2H),3.81(s,1H),2.41(s,2H),1.49(d,J=6.6Hz,3H).

[0571] Example 41

[0572] (4-((4-(3-((2-((S)-1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(tetrahydrofuran-3-yl)methanone

[0573] (4-((4-(3-((2-((S)-1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(tetrahydrofuran-3-yl)methanone

[0574] first step

[0575] (4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(tetrahydrofuran-3-yl)methanone

[0576] (4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(tetrahydrofuran-3-yl)methanone

[0577] Tetrahydrofuran-3-carboxylic acid 41a (20.26 mg, 174.46 μmol, commercially available), 3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)-5-(4-((1,2,3,6-tetrahydropyridin-4-yl)ethynyl)phenyl)isoxazole 35f (40 mg, 87.23 μmol), 1-hydroxybenzotriazole (23.57 mg, 174.46 μmol), N-methylmorpholine (26.47 mg, 261.69 μmol), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (33.44 mg, 174.46 μmol) were added sequentially to dichloromethane (1.5 mL) and stirred at room temperature for 16 hours. After the reaction was complete, water (50 mL) was added, and dichloromethane (50 mL×3) was added for extraction. The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to give (4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(tetrahydrofuran-3-yl)methanone 41b (48 mg) in a yield of 98.85%. The crude product was used directly in the next step.

[0578] MS m / z(ESI):557.3[M+1].

[0579] Step 2

[0580] (4-((4-(3-((2-((S)-1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(tetrahydrofuran-3-yl)methanone

[0581] (4-((4-(3-((2-((S)-1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(tetrahydrofuran-3-yl)methanone

[0582] (4-((4-(3-((2-((1S)-1-((tetrahydro2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(tetrahydrofuran-3-yl)methanone 41b (48 mg, 86.23 μmol) was added to dichloromethane (1.5 mL), followed by the dropwise addition of trifluoroacetic acid (0.5 mL), and stirring was continued at room temperature for 1 hour. The crude product was concentrated and subjected to preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H2O, mobile phase B: CH3CN) to give (4-((4-(3-((2-((S)-1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(tetrahydrofuran-3-yl)methanone 41 (20.51 mg) in a yield of 36.90%.

[0583] MS m / z(ESI):473.3[M+1].

[0584] Example 42

[0585] (4-((4-(3-((2-((S)-1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(morpholin-3-yl)methanone

[0586] (4-((4-(3-((2-((S)-1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(morpholin-3-yl)methanone

[0587] first step

[0588] morpholin-3-yl(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0589] Morpholin-3-yl(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0590] Morpholine-3-carboxylic acid 42a (22.88 mg, 174.46 μmol, commercially available), 3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)-5-(4-((1,2,3,6-tetrahydropyridin-4-yl)ethynyl)phenyl)isoxazole 35f (40 mg, 87.23 μmol), 1-hydroxybenzotriazole (23.57 mg, 174.46 μmol), N-methylmorpholine (26.47 mg, 261.69 μmol), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (33.44 mg, 174.46 μmol) were added sequentially to dichloromethane (1.5 mL) and stirred at room temperature for 16 hours. After the reaction was complete, water (50 mL) was added, followed by extraction with dichloromethane (50 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to afford morpholin-3-yl(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone 42b (45 mg) in a 90.24% yield. The crude product was used directly in the next reaction.

[0591] MS m / z(ESI):572.3[M+1].

[0592] Step 2

[0593] (4-((4-(3-((2-((S)-1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(morpholin-3-yl)methanone

[0594] (4-((4-(3-((2-((S)-1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(morpholin-3-yl)methanone

[0595] Morpholin-3-yl(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone 42b (45 mg, 78.72 μmol) was added to dichloromethane (1.5 mL), followed by the dropwise addition of trifluoroacetic acid (0.5 mL), and stirring was continued at room temperature for 1 hour. The crude product was concentrated and subjected to preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H2O, mobile phase B: CH3CN) to give (4-((4-(3-((2-((S)-1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(morpholin-3-yl)methanone 42 (10.35 mg) in a yield of 20.33%.

[0596] MS m / z(ESI):488.3[M+1].

[0597] Example 43

[0598] (S)-1-(1-((5-(4-((1-(2-(1H-1,2,4-triazol-1-yl)ethyl)ethyl)-1,2,3,6-tetrahydropyridin-4-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol

[0599] (S)-1-(1-((5-(4-((1-(2-(1H-1,2,4-triazol-1-yl)ethyl)-1,2,3,6-tetrahydropyridin-4-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol

[0600] first step

[0601] 5-(4-((1-(2-(1H-1,2,4-triazol-1-yl)ethyl)-1,2,3,6-tetrahydropyridin-4-yl)ethynyl)phenyl )-3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazole

[0602] 5-(4-((1-(2-(1H-1,2,4-triazol-1-yl)ethyl)-1,2,3,6-tetrahydropyridin-4-yl)ethynyl)phenyl)-3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazole

[0603] Ethyl 2-(1H-1,2,4-triazol-1-yl)methanesulfonate 43a (25.02 mg, 130.85 μmol), 3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)-5-(4-((1,2,3,6-tetrahydropyridin-4-yl)ethynyl)phenyl)isoxazole 35f (30 mg, 65.42 μmol) and potassium carbonate (9.04 mg, 65.42 μmol) were added to acetonitrile (2 mL) in sequence and stirred at 60 °C for 16 hours. After the reaction was complete, water (50 mL) was added, followed by extraction with dichloromethane (50 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to afford 5-(4-((1-(2-(1H-1,2,4-triazol-1-yl)ethyl)-1,2,3,6-tetrahydropyridin-4-yl)ethynyl)phenyl)-3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazole 43b (35 mg) in a 96.63% yield. The crude product was used directly in the next reaction.

[0604] MS m / z(ESI):554.4[M+1].

[0605] Step 2

[0606] (S)-1-(1-((5-(4-((1-(2-(1H-1,2,4-triazol-1-yl)ethyl)ethyl)-1,2,3,6-tetrahydropyridin-4-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol

[0607] (S)-1-(1-((5-(4-((1-(2-(1H-1,2,4-triazol-1-yl)ethyl)-1,2,3,6-tetrahydropyridin-4-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol

[0608] 5-(4-((1-(2-(1H-1,2,4-triazol-1-yl)ethyl)-1,2,3,6-tetrahydropyridin-4-yl)ethynyl)phenyl)-3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazole 43b (35 mg, 63.22 μmol) was added to dichloromethane (1.5 mL), followed by the dropwise addition of trifluoroacetic acid (0.5 mL), and stirring was continued at room temperature for 1 hour. The crude product was concentrated and subjected to preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H2O, mobile phase B: CH3CN) to give (S)-1-(1-((5-(4-((1-(2-(1H-1,2,4-triazol-1-yl)ethyl)-1,2,3,6-tetrahydropyridin-4-yl)ethynyl)phenyl)isoxazol-3-yl)methyl)-1H-imidazol-2-yl)ethan-1-ol 43 (10.69 mg) in a yield of 28.98%.

[0609] MS m / z(ESI):470.3[M+1].

[0610] Example 44

[0611] (S)-3-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)propanenitrile

[0612] (S)-3-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)propionitrile

[0613] first step

[0614] 3-(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)propanenitrile

[0615] 3-(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)propionitrile

[0616] Acrylonitrile 44a (13.89 mg, 261.69 μmol, commercially available), 3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)-5-(4-((1,2,3,6-tetrahydropyridin-4-yl)ethynyl)phenyl)isoxazole 35f (60 mg, 130.85 μmol), and potassium carbonate (36.17 mg, 261.69 μmol) were added to acetonitrile (1 mL) and water (0.5 mL) in sequence and stirred continuously at room temperature for 16 h. After the reaction was complete, water (50 mL) was added, followed by extraction with dichloromethane (50 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to afford 3-(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)propionitrile 44b (66 mg) in a 98.59% yield. The crude product was used directly in the next reaction.

[0617] MS m / z(ESI):512.3[M+1].

[0618] Step 2

[0619] (S)-3-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)propanenitrile

[0620] (S)-3-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)propionitrile

[0621] 3-(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)propionitrile 44b (66 mg, 129.00 μmol) was added to dichloromethane (1.5 mL), followed by the dropwise addition of trifluoroacetic acid (0.5 mL), and stirring was continued at room temperature for 1 hour. The crude product was concentrated and subjected to preparative liquid separation (AKZONOBEL Kromasil column; 250 × 21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA + H₂O, mobile phase B: CH₃CN) to afford (S)-3-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)propionitrile 44 (32.38 mg) in a yield of 46.35%. MS m / z (ESI): 428.2 [M+1].

[0622] Example 45

[0623] (S)-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(pyridin-3-yl)methanone

[0624] (S)-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(pyridin-3-yl)methanone

[0625] first step

[0626] pyridin-3-yl(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0627] Pyridin-3-yl(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0628] Nicotinic acid 45a (21.48 mg, 174.46 μmol, commercially available), 3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)-5-(4-((1,2,3,6-tetrahydropyridin-4-yl)ethynyl)phenyl)isoxazole 35f (40 mg, 87.23 μmol), 1-hydroxybenzotriazole (23.57 mg, 174.46 μmol), N-methylmorpholine (26.47 mg, 261.69 μmol), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (33.44 mg, 174.46 μmol) were added sequentially to dichloromethane (1.5 mL) and stirred at room temperature for 16 hours. After the reaction was complete, water (50 mL) was added, followed by extraction with dichloromethane (50 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to afford pyridin-3-yl(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone 45b (49 mg) in a 99.66% yield. The crude product was used directly in the next reaction.

[0629] MS m / z(ESI):564.3[M+1].

[0630] Step 2

[0631] (S)-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(pyridin-3-yl)methanone

[0632] (S)-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(pyridin-3-yl)methanone

[0633] Pyridin-3-yl(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone 45b (49 mg, 86.93 μmol) was added to dichloromethane (1.5 mL), followed by the dropwise addition of trifluoroacetic acid (0.5 mL), and stirring was continued at room temperature for 1 hour. The crude product was concentrated and subjected to preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H2O, mobile phase B: CH3CN) to give (S)-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(pyridin-3-yl)methanone 45 (37.08 mg) in a yield of 70.42%.

[0634] MS m / z(ESI):480.2[M+1].

[0635] Examples 46-56 were synthesized according to the synthesis method of Examples 1-45 of the present invention, and their structures and characterization data are shown in the following table:

[0636] Example 57

[0637] (S)-4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridine-1(2H)-carbaldehyde

[0638] (S)-4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridine-1(2H)-carbaldehyde

[0639] first step

[0640] 4-((trimethylsilyl)ethynyl)-3,6-dihydropyridine-1(2H)-carbaldehyde

[0641] 4-((Trimethylsilyl)ethynyl)-3,6-dihydropyridine-1(2H)-carbaldehyde

[0642] 4-((Trimethylsilyl)ethynyl)-1,2,3,6-tetrahydropyridine 57a (250 mg, 1.16 mmol, commercially available) and triethylamine (468.92 mg, 4.63 mmol, 642.35 μL) were added sequentially to formamide (5.13 mL) and stirred at room temperature for 12 hours. Ethyl acetate (30 mL) and water (15 mL) were added, the organic phase was separated, and the aqueous phase was extracted with ethyl acetate (30 mL × 2). The combined organic phases were washed with saturated sodium chloride solution (30 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated to dryness under reduced pressure to obtain 4-((trimethylsilyl)ethynyl)-3,6-dihydropyridine-1(2H)-carbaldehyde 57b (220 mg), which was used directly in the next reaction.

[0643] MS m / z(ESI):208.0[M+1].

[0644] Step 2

[0645] 4-ethynyl-3,6-dihydropyridine-1(2H)-carbaldehyde

[0646] 4-Ethynyl-3,6-dihydropyridine-1(2H)-carbaldehyde

[0647] 4-((Trimethylsilyl)ethynyl)-3,6-dihydropyridine-1(2H)-carbaldehyde 57b (220 mg, 1.06 mmol) and potassium fluoride (308.23 mg, 5.31 mmol) were added sequentially to methanol (5 mL) and stirred at room temperature for 3 hours. The mixture was concentrated to dryness under reduced pressure, dissolved in acetonitrile (3 mL), filtered, and the filtrate was concentrated to dryness under reduced pressure to afford 4-ethynyl-3,6-dihydropyridine-1(2H)-carbaldehyde 57c (140 mg), which was used directly in the next reaction.

[0648] MS m / z(ESI):136.0[M+1].

[0649] Step 3

[0650] 4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridine-1(2H)-carbaldehyde

[0651] 4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridine-1(2H)-carbaldehyde

[0652] 5-(4-Iodophenyl)-3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazole 1i (250 mg, 495.50 μmol), 4-ethynyl-3,6-dihydropyridine-1(2H)-carbaldehyde 57c (118.19 mg, 743.26 μmol), triethylenediamine (111.16 mg, 991.01 μmol) and allylpalladium(II) chloride dimer (36.17 mg, 99.10 μmol) were added sequentially to acetonitrile (2.0 mL), the argon atmosphere was replaced three times, and stirring was continued at room temperature for 3 hours. Ethyl acetate (30 mL) and water (15 mL) were added, and the mixture was separated. The aqueous phase was extracted with ethyl acetate (30 mL × 2). The combined organic phases were washed with saturated sodium chloride solution (30 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated to dryness under reduced pressure. The residue was separated and purified by silica gel column chromatography (eluent: System A) to obtain 4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridine-1(2H)-carbaldehyde 57d (240 mg) in an 89.59% yield. MS m / z (ESI): 487.3 [M+1].

[0653] Step 4

[0654] (S)-4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridine-1(2H)-carbaldehyde

[0655] (S)-4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridine-1(2H)-carbaldehyde

[0656] 4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridine-1(2H)-carbaldehyde 57d (220 mg, 452.15 μmol) and trifluoroacetic acid (0.5 mL) were added to dichloromethane (2 mL), and stirring was continued at room temperature for 1 hour. The crude product was concentrated and subjected to preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H2O, mobile phase B: CH3CN) to give (S)-4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridine-1(2H)-carbaldehyde 57 (59.7 mg) in a yield of 25.05%.

[0657] MS m / z(ESI):402.9[M+1]. 1 H NMR (400MHz, DMSO-d6) δ8.09 (s, 1H), 7.91–7.85 (m, 2H), 7.79 (d, J = 2.0Hz, 1H), 7.71(d,J=2.0Hz,1H),7.61(d,J=8.2Hz,2H),7.18(s,1H),6.32–6.21(m,1H),5. 73(s,2H),5.27(q,J=6.6Hz,1H),4.04(dq,J=20.9,3.0Hz,2H),3.55(t,J=5.7Hz ,2H),2.36(dq,J=5.8,2.9Hz,1H),2.26(h,J=2.7Hz,1H),1.50(d,J=6.6Hz,3H).

[0658] Example 58

[0659] (S)-4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridine-1(2H)-carboxamide

[0660] (S)-4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridine-1(2H)-carboxamide

[0661] first step

[0662] 4-((trimethylsilyl)ethynyl)-3,6-dihydropyridine-1(2H)-carboxamide

[0663] 4-((Trimethylsilyl)ethynyl)-3,6-dihydropyridine-1(2H)-carboxamide

[0664] 4-((Trimethylsilyl)ethynyl)-1,2,3,6-tetrahydropyridine 57a (50 mg, 278.81 μmol) and triethylamine (84.64 mg, 836.43 μmol, 115.94 μL) were added sequentially to tetrahydrofuran (1.95 mL), followed by isocyanatotrimethylsilane 58a (41.76 mg, 362.45 μmol, 49.13 μL, commercially available), and the mixture was allowed to react at room temperature for 16 hours. After completion of the reaction, the mixture was concentrated to afford 4-((trimethylsilyl)ethynyl)-3,6-dihydropyridine-1(2H)-carboxamide 58b (50 mg), which was used directly in the next step.

[0665] MS m / z(ESI):223.0[M+1].

[0666] Step 2

[0667] 4-ethynyl-3,6-dihydropyridine-1(2H)-carboxamide

[0668] 4-Ethynyl-3,6-dihydropyridine-1(2H)-carboxamide

[0669] 4-((Trimethylsilyl)ethynyl)-3,6-dihydropyridine-1(2H)-carboxamide 58b (50 mg, 224.86 μmol) and potassium fluoride (65.32 mg, 1.12 mmol) were added sequentially to methanol (2 mL) and stirred at room temperature for 12 hours. The system was concentrated to dryness under reduced pressure, and ethyl acetate (30 mL) and water (15 mL) were added. The organic phase was separated, and the aqueous phase was extracted with ethyl acetate (30 mL × 2). The combined organic phases were washed with saturated sodium chloride solution (30 mL × 2) and concentrated under reduced pressure to afford 4-ethynyl-3,6-dihydropyridine-1(2H)-carboxamide 58c (20 mg), which was used directly in the next reaction.

[0670] MS m / z(ESI):151.1[M+1].

[0671] Step 3

[0672] 4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridine-1(2H)-carboxamide

[0673] 4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridine-1(2H)-carboxamide

[0674] Under argon atmosphere, 5-(4-iodophenyl)-3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazole 1i (50 mg, 104.32 μmol), 4-ethynyl-3,6-dihydropyridine-1(2H)-carboxamide 58c (23.50 mg, 156.47 μmol), triethylenediamine (23.40 mg, 208.63 μmol), allylpalladium(II) chloride dimer (7.62 mg, 20.86 μmol), and tri-tert-butylphosphine (10% in toluene solution) (42.21 mg, 20.86 μmol, 10% purity) were added sequentially to acetonitrile (2 mL). After argon replacement, stirring was continued at room temperature for 4 hours. Ethyl acetate (30 mL) and water (15 mL) were added, and the layers were separated. The aqueous phase was extracted with ethyl acetate (30 mL × 2). The combined organic phases were washed with saturated sodium chloride solution (30 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated to dryness under reduced pressure. The residue was separated and purified by silica gel chromatography (eluent: System A) to give 4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridine-1(2H)-carboxamide 58d (40 mg) in a yield of 76.45%.

[0675] MS m / z(ESI):502.3[M+1].

[0676] Step 4

[0677] (S)-4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridine-1(2H)-carboxamide

[0678] (S)-4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridine-1(2H)-carboxamide

[0679] 4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridine-1(2H)-carboxamide 58d (40 mg, 79.75 μmol) and trifluoroacetic acid (0.5 mL) were added sequentially to dichloromethane (2 mL), and stirring was continued at room temperature for 1 hour. The crude product was concentrated and subjected to preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H2O, mobile phase B: CH3CN) to give (S)-4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridine-1(2H)-carboxamide 58 (13.5 mg) in a yield of 30.26%.

[0680] MS m / z(ESI):418.2[M+1].

[0681] Example 59

[0682] (S)-2-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)acetamide

[0683] (S)-2-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)acetamide

[0684] first step

[0685] 2-(4-((trimethylsilyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)acetamide

[0686] 2-(4-((Trimethylsilyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)acetamide

[0687] 2-Iodoacetamide 59a (123.77 mg, 669.14 μmol, commercially available), 4-((trimethylsilyl)ethynyl)-1,2,3,6-tetrahydropyridine 57a (100 mg, 557.62 μmol), and cesium carbonate (363.37 mg, 1.12 mmol) were added sequentially to acetonitrile (2 mL). The mixture was stirred at room temperature for 12 hours. The organic solvent was removed by distillation under reduced pressure. Ethyl acetate (50 mL) and water (30 mL) were then added. The layers were separated, and the aqueous phase was extracted with ethyl acetate (30 mL × 2). The organic phases were combined, dried over anhydrous sodium sulfate, and then the organic solvent was removed by distillation under reduced pressure to obtain 2-(4-((trimethylsilyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)acetamide 59b (130 mg), which was used directly in the next reaction.

[0688] MS m / z(ESI):237.2[M+1].

[0689] Step 2

[0690] 2-(4-ethynyl-3,6-dihydropyridin-1(2H)-yl)acetamide

[0691] 2-(4-ethynyl-3,6-dihydropyridin-1(2H)-yl)acetamide

[0692] 2-(4-((Trimethylsilyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)acetamide 59b (130 mg, 549.95 μmol) and potassium fluoride (95.86 mg, 1.65 mmol) were added to methanol (2 mL) in sequence. The mixture was stirred at room temperature for 16 hours. The organic solvent was removed by distillation under reduced pressure to obtain 2-(4-ethynyl-3,6-dihydropyridin-1(2H)-yl)acetamide 59c (80 mg), which was used directly in the next reaction.

[0693] MS m / z(ESI):165.2[M+1].

[0694] Step 3

[0695] 2-(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)acetamide

[0696] 2-(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)acetamide

[0697] 5-(4-iodophenyl)-3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazole 1i (150 mg, 312.95 μmol), 2-(4-ethynyl-3,6-dihydropyridin-1(2H)-yl)acetamide 59c (56.53 mg, 344.24 μmol), triethylenediamine (70.21 mg, 625.90 μmol), allylpalladium(II) chloride dimer (22.85 mg, 62.59 μmol) and tri-tert-butylphosphine (10% in toluene solution) (12.66 mg, 62.59 μmol) were added sequentially to acetonitrile (2 mL). After argon replacement, stirring was continued at room temperature for 4 hours. The reaction solution was distilled under reduced pressure to remove the organic solvent, and the residue was separated and purified by silica gel column chromatography (eluent: System A) to give 2-(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)acetamide 59d (108 mg) in a yield of 66.93%.

[0698] MS m / z(ESI):516.3[M+1].

[0699] Step 4

[0700] (S)-2-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)acetamide

[0701] (S)-2-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)acetamide

[0702] 2-(4-((4-(3-((2-((1S)-1-((tetrahydro2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)acetamide 59d (50 mg, 96.97 μmol) and trifluoroacetic acid (0.5 mL) were added sequentially to dichloromethane (1.5 mL), and stirring was continued at room temperature for 3 hours. The crude product was concentrated and subjected to preparative liquid separation (AKZONOBEL Kromasil column; 250 × 21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA + H₂O, mobile phase B: CH₃CN) to afford (S)-2-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)acetamide 59 (26.0 mg) in a 43.60% yield. MS m / z (ESI): 431.9 [M+1].

[0703] Example 60

[0704] (S)-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(morpholino)methanone

[0705] (S)-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(morpholino)methanone

[0706] first step

[0707] morpholino(4-((trimethylsilyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0708] Morpholino(4-((trimethylsilyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0709] 4-((Trimethylsilyl)ethynyl)-1,2,3,6-tetrahydropyridine 57a (62.50 mg, 278.81 μmol), morpholine 60a (48.58 mg, 557.62 μmol, 49.07 μL), triphosgene (99.28 mg, 334.57 μmol) and triethylamine (169.28 mg, 1.67 mmol, 235.11 μL) were added sequentially to tetrahydrofuran (1.72 mL) and stirred at room temperature for 12 hours. Ethyl acetate (30 mL) and water (15 mL) were added, the organic phase was separated, and the aqueous phase was extracted with ethyl acetate (30 mL × 2). The combined organic phases were washed with saturated sodium chloride solution (30 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated to dryness under reduced pressure. The residue was separated and purified by silica gel column chromatography (eluent: System A) to give morpholino(4-((trimethylsilyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone 60b (40 mg) in a yield of 49.06%.

[0710] MS m / z(ESI):293.0[M+1].

[0711] Step 2

[0712] (4-ethynyl-3,6-dihydropyridin-1(2H)-yl)(morpholino)methanone

[0713] (4-Ethynyl-3,6-dihydropyridin-1(2H)-yl)(morpholino)methanone

[0714] Morpholino(4-((trimethylsilyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone 60b (40 mg, 136.78 μmol) and potassium fluoride (39.73 mg, 683.88 μmol) were added sequentially to methanol (1 mL) and stirred at room temperature for 4 hours. The system was concentrated to dryness under reduced pressure, and acetonitrile (10 mL) was added to the residue, filtered, and the filtrate was concentrated to dryness to afford (4-ethynyl-3,6-dihydropyridin-1(2H)-yl)(morpholino)methanone 60c (20 mg), which was used directly in the next reaction.

[0715] MS m / z(ESI):221.0[M+1].

[0716] Step 3

[0717] morpholino(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0718] Morpholino(4-((4-(3-((2-((1S)-1-((tetrahydro2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0719] 5-(4-iodophenyl)-3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazole 1i (25 mg, 49.55 μmol), (4-ethynyl-3,6-dihydropyridin-1(2H)-yl)(morpholino)methanone 60c (18.19 mg, 74.33 μmol), triethylenediamine (11.12 mg, 99.10 μmol) and allylpalladium(II) chloride dimer (3.62 mg, 9.91 μmol) were added sequentially to acetonitrile (2 mL), argon was replaced three times, and the mixture was stirred at room temperature for 12 hours. The system was concentrated to dryness under reduced pressure, and the residue was separated and purified by silica gel column chromatography (eluent: System A) to give morpholino(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone 60d (20 mg) in a yield of 70.61%.

[0720] MS m / z(ESI):572.0[M+1].

[0721] Step 4

[0722] (S)-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(morpholino)methanone

[0723] (S)-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(morpholino)methanone

[0724] Morpholino(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone 60d (20 mg, 34.99 μmol) and trifluoroacetic acid (0.3 mL) were added sequentially to dichloromethane (1 mL), and stirring was continued at room temperature for 1 hour. The system was concentrated to dryness under reduced pressure, and the residue was subjected to preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H 2 O, mobile phase B: CH 3 CN) to give (S)-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(morpholino)methanone 60 (2.5 mg) in a yield of 11.11%.

[0725] MS m / z(ESI):487.9[M+1].

[0726] Example 61

[0727] (S)-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(pyridin-2-yl)methanone

[0728] (S)-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(pyridin-2-yl)methanone

[0729] first step

[0730] pyridin-2-yl(4-((trimethylsilyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0731] Pyridin-2-yl(4-((trimethylsilyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0732] 4-((Trimethylsilyl)ethynyl)-1,2,3,6-tetrahydropyridine 57a (50 mg, 278.81 μmol), picolinic acid 61a (51.49 mg, 418.22 μmol, commercially available), O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (127.21 mg, 334.57 μmol), and N,N-diisopropylethylamine (108.10 mg, 836.43 μmol, 145.69 μL) were added sequentially to dichloromethane (2.0 mL) and stirred at room temperature for 4 hours. Mass spectrometry indicated that the reaction of the starting material was complete, and the system was concentrated to dryness under reduced pressure to obtain pyridin-2-yl(4-((trimethylsilyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone 61b (55 mg), which was used directly in the next reaction.

[0733] MS m / z(ESI):285.1[M+1].

[0734] Step 2

[0735] (4-ethynyl-3,6-dihydropyridin-1(2H)-yl)(pyridin-2-yl)methanone

[0736] (4-Ethynyl-3,6-dihydropyridin-1(2H)-yl)(pyridin-2-yl)methanone

[0737] Pyridin-2-yl(4-((trimethylsilyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone 61b (55 mg, 193.37 μmol) and potassium fluoride (56.17 mg, 966.85 μmol) were added sequentially to methanol (2 mL) and stirred at room temperature for 3 hours. The system was concentrated to dryness under reduced pressure, dissolved in acetonitrile (3 mL), filtered, and the filtrate was concentrated to dryness under reduced pressure to afford (4-ethynyl-3,6-dihydropyridin-1(2H)-yl)(pyridin-2-yl)methanone 61c (30 mg), which was used directly in the next reaction.

[0738] MS m / z(ESI):213.0[M+1].

[0739] Step 3

[0740] pyridin-2-yl(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0741] Pyridin-2-yl(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0742] 5-(4-iodophenyl)-3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazole 1i (30 mg, 59.46 μmol), (4-ethynyl-3,6-dihydropyridin-1(2H)-yl)(pyridin-2-yl)methanone 61c (21.03 mg, 89.19 μmol), triethylenediamine (13.34 mg, 118.92 μmol) and allylpalladium(II) chloride dimer (4.34 mg, 11.89 μmol) were added sequentially to acetonitrile (2 mL), argon was replaced three times, and the mixture was stirred at room temperature for 12 hours. The system was concentrated to dryness under reduced pressure, and the residue was separated and purified by silica gel column chromatography (eluent: System A) to give pyridin-2-yl(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone 61d (25 mg) in a yield of 74.59%.

[0743] MS m / z(ESI):563.9[M+1].

[0744] Step 4

[0745] (S)-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(pyridin-2-yl)methanone

[0746] (S)-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(pyridin-2-yl)methanone

[0747] Pyridin-2-yl(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone 61d (25 mg, 44.35 μmol) and trifluoroacetic acid (0.5 mL) were added sequentially to dichloromethane (2 mL), and stirring was continued at room temperature for 1 hour. The system was concentrated to dryness under reduced pressure, and the residue was subjected to preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H2O, mobile phase B: CH3CN) to give (S)-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(pyridin-2-yl)methanone 61 (15.2 mg) in a yield of 56.00%.

[0748] MS m / z(ESI):480.2[M+1].

[0749] Example 62

[0750] (S)-1-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)ethan-1-one

[0751] (S)-1-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)ethan-1-one

[0752] first step

[0753] 1-(4-((trimethylsilyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)ethan-1-one

[0754] 1-(4-((Trimethylsilyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)ethan-1-one

[0755] 4-((Trimethylsilyl)ethynyl)-1,2,3,6-tetrahydropyridine 57a (50 mg, 278.81 μmol), acetic anhydride (42.70 mg, 418.22 μmol, 39.53 μL), and triethylamine (84.64 mg, 836.43 μmol, 116.26 μL) were added sequentially to dichloromethane (1 mL) and stirred at room temperature for 12 hours. The system was concentrated to dryness under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: System A) to afford 1-(4-((trimethylsilyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)ethan-1-one 62a (50 mg) in an 81.01% yield.

[0756] MS m / z(ESI):222.0[M+1].

[0757] Step 2

[0758] 1-(4-ethynyl-3,6-dihydropyridin-1(2H)-yl)ethan-1-one

[0759] 1-(4-Ethynyl-3,6-dihydropyridin-1(2H)-yl)ethan-1-one

[0760] 1-(4-((Trimethylsilyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)ethan-1-one 62a (50 mg, 225.87 μmol) and potassium fluoride (65.61 mg, 1.13 mmol) were added sequentially to methanol (1 mL) and stirred at room temperature for 3 hours. The mixture was concentrated to dryness under reduced pressure, dissolved in acetonitrile (3 mL), filtered, and the filtrate was concentrated to dryness under reduced pressure to afford 1-(4-ethynyl-3,6-dihydropyridin-1(2H)-yl)ethan-1-one 62b (30 mg), which was used directly in the next reaction.

[0761] MS m / z(ESI):150.0[M+1].

[0762] Step 3

[0763] 1-(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)ethan-1-one

[0764] 1-(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)ethan-1-one

[0765] 5-(4-iodophenyl)-3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazole 1i (30 mg, 59.46 μmol), 1-(4-ethynyl-3,6-dihydropyridin-1(2H)-yl)ethan-1-one 62b (14.78 mg, 89.19 μmol), triethylenediamine (13.34 mg, 118.92 μmol) and allylpalladium(II) chloride dimer (4.34 mg, 11.89 μmol) were added sequentially to acetonitrile (2 mL), argon was replaced three times, and the mixture was stirred at room temperature for 12 hours. The system was concentrated to dryness under reduced pressure, and the residue was separated and purified by silica gel column chromatography (eluent: System A) to give 1-(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)ethan-1-one 62c (25 mg) in a yield of 83.99%.

[0766] MS m / z(ESI):500.9[M+1].

[0767] Step 4

[0768] (S)-1-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)ethan-1-one

[0769] (S)-1-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)ethan-1-one

[0770] 1-(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)ethan-1-one 62c (25 mg, 49.94 μmol) and trifluoroacetic acid (0.5 mL) were added sequentially to dichloromethane (2 mL), and stirring was continued at room temperature for 1 hour. The system was concentrated to dryness under reduced pressure, and the residue was subjected to preparative liquid separation (AKZONOBEL Kromasil column; 250 × 21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA + H₂O, mobile phase B: CH₃CN) to afford (S)-1-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)ethan-1-one 62 (6.8 mg) in a yield of 24.38%. MS m / z (ESI): 417.2 [M+1].

[0771] Example 63

[0772] (S)-2-hydroxy-1-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)ethan-1-one

[0773] (S)-2-Hydroxy-1-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)ethan-1-one

[0774] first step

[0775] 2-hydroxy-1-(4-((trimethylsilyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)ethan-1-one

[0776] 2-Hydroxy-1-(4-((trimethylsilyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)ethan-1-one

[0777] 4-((Trimethylsilyl)ethynyl)-1,2,3,6-tetrahydropyridine 57a (50 mg, 278.81 μmol), 2-hydroxyacetic acid 12a (42.41 mg, 557.62 μmol), O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (127.21 mg, 334.57 μmol), and N,N-diisopropylethylamine (108.10 mg, 836.43 μmol, 145.69 μL) were added sequentially to dichloromethane (2.0 mL) and stirred at room temperature for 4 hours. Mass spectrometry indicated that the reaction of the starting materials was complete, and the system was concentrated to dryness under reduced pressure to obtain 2-hydroxy-1-(4-((trimethylsilyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)ethan-1-one 63a (40 mg), which was used directly in the next reaction.

[0778] MS m / z(ESI):237.9[M+1].

[0779] Step 2

[0780] 1-(4-ethynyl-3,6-dihydropyridin-1(2H)-yl)-2-hydroxyethan-1-one

[0781] 1-(4-Ethynyl-3,6-dihydropyridin-1(2H)-yl)-2-hydroxyethan-1-one

[0782] 2-Hydroxy-1-(4-((trimethylsilyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)ethan-1-one 63a (40 mg, 168.51 μmol) and potassium fluoride (48.95 mg, 842.57 μmol) were added sequentially to methanol (1 mL) and stirred at room temperature for 3 hours. The solution was concentrated to dryness under reduced pressure, dissolved in acetonitrile (3 mL), filtered, and the filtrate was concentrated to dryness under reduced pressure to afford 1-(4-ethynyl-3,6-dihydropyridin-1(2H)-yl)-2-hydroxyethan-1-one 63b (20 mg), which was used directly in the next reaction.

[0783] MS m / z(ESI):166.0[M+1].

[0784] Step 3

[0785] 2-hydroxy-1-(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)ethan-1-one

[0786] 2-Hydroxy-1-(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)ethan-1-one

[0787] 5-(4-iodophenyl)-3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazole 1i (30 mg, 59.46 μmol), 1-(4-ethynyl-3,6-dihydropyridin-1(2H)-yl)-2-hydroxyethan-1-one 63b (16.37 mg, 89.19 μmol), triethylenediamine (13.34 mg, 118.92 μmol) and allylpalladium(II) chloride dimer (4.34 mg, 11.89 μmol) were added sequentially to acetonitrile (2 mL), argon was replaced three times, and the mixture was stirred at room temperature for 12 hours. The system was concentrated to dryness under reduced pressure, and the residue was separated and purified by silica gel column chromatography (eluent: System A) to give 2-hydroxy-1-(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)ethan-1-one 63c (25 mg) in a yield of 81.39%.

[0788] MS m / z(ESI):516.9[M+1].

[0789] Step 4

[0790] (S)-2-hydroxy-1-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)ethan-1-one

[0791] (S)-2-Hydroxy-1-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)ethan-1-one

[0792] 2-Hydroxy-1-(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)ethan-1-one 63c (25 mg, 48.39 μmol) and trifluoroacetic acid (0.5 mL) were added sequentially to dichloromethane (2 mL), and stirring was continued at room temperature for 1 hour. The system was concentrated to dryness under reduced pressure, and the residue was subjected to preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H 2 O, mobile phase B: CH 3 CN) to give (S)-2-hydroxy-1-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)ethan-1-one 63 (1.3 mg) in a yield of 4.62%.

[0793] MS m / z(ESI):433.2[M+1]. 1 H NMR(400MHz,DMSO-d6)δ7.89–7.86(m,2H),7.76–7.75(d,J=4.0Hz,1H),7.6 7–7.66(d,J=4.0Hz,1H),7.62–7.60(m,3H),7.17–7.16(d,J=4.0Hz,1H),6. 27–6.24(m,1H),5.73(s,2H),5.24(q,J=6.6Hz,1H),4.41–4.04(m,4H),3.6 4–3.61(m,1H),3.50–3.47(m,1H),2.35–2.28(m,2H),1.50(d,J=6.6Hz,3H).

[0794] Example 64

[0795] (1,4-dioxan-2-yl)(4-((4-(3-((2-((S)-1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0796] (1,4-Dioxan-2-yl)(4-((3-((2-((S)-1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0797] first step

[0798] (1,4-dioxan-2-yl)(4-((trimethylsilyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0799] (1,4-Dioxan-2-yl)(4-((trimethylsilyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0800] 4-((Trimethylsilyl)ethynyl)-1,2,3,6-tetrahydropyridine 57a (50 mg, 278.81 μmol), 1,4-dioxane-2-carboxylic acid 64a (55.25 mg, 418.22 μmol), O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (127.21 mg, 334.57 μmol), and N,N-diisopropylethylamine (108.10 mg, 836.43 μmol, 145.69 μL) were added sequentially to dichloromethane (2.0 mL) and stirred at room temperature for 4 hours. Mass spectrometry indicated that the reaction of the starting materials was complete, and the system was concentrated to dryness under reduced pressure to obtain (1,4-dioxane-2-yl)(4-((trimethylsilyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone 64b (45 mg), which was used directly in the next reaction.

[0801] MS m / z(ESI):294.0[M+1].

[0802] Step 2

[0803] (1,4-dioxan-2-yl)(4-ethynyl-3,6-dihydropyridin-1(2H)-yl)methanone

[0804] (1,4-Dioxan-2-yl)(4-ethynyl-3,6-dihydropyridin-1(2H)-yl)methanone

[0805] (1,4-Dioxan-2-yl)(4-((trimethylsilyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone 64b (45 mg, 153.36 μmol) and potassium fluoride (8.91 mg, 153.36 μmol) were added sequentially to methanol (2 mL) and stirred at room temperature for 3 hours. The system was concentrated to dryness under reduced pressure, dissolved in acetonitrile (3 mL), filtered, and the filtrate was concentrated to dryness under reduced pressure to give (1,4-dioxan-2-yl)(4-ethynyl-3,6-dihydropyridin-1(2H)-yl)methanone 64c (25 mg), which was used directly in the next reaction.

[0806] MS m / z(ESI):222.1[M+1].

[0807] Step 3

[0808] (1,4-dioxan-2-yl)(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0809] (1,4-Dioxan-2-yl)(4-((3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0810] 5-(4-iodophenyl)-3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazole 1i (30 mg, 59.46 μmol), (1,4-dioxan-2-yl)(4-ethynyl-3,6-dihydropyridin-1(2H)-yl)methanone 64c (21.93 mg, 89.19 μmol), triethylenediamine (13.34 mg, 118.92 μmol) and allylpalladium(II) chloride dimer (4.34 mg, 11.89 μmol) were added sequentially to acetonitrile (2 mL), argon was replaced three times, and the mixture was stirred at room temperature for 12 hours. The system was concentrated to dryness under reduced pressure, and the residue was separated and purified by silica gel column chromatography (eluent: System A) to give (1,4-dioxane-2-yl)(4-((3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone 64d (25 mg) in a yield of 73.42%.

[0811] MS m / z(ESI):573.0[M+1].

[0812] Step 4

[0813] (1,4-dioxan-2-yl)(4-((4-(3-((2-((S)-1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0814] (1,4-Dioxan-2-yl)(4-((3-((2-((S)-1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0815] (1,4-Dioxan-2-yl)(4-((3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone 64d (25 mg, 43.66 μmol) and trifluoroacetic acid (0.5 mL) were added sequentially to dichloromethane (2 mL), and stirring was continued at room temperature for 1 hour. The system was concentrated to dryness under reduced pressure, and the residue was subjected to preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H2O, mobile phase B: CH3CN) to give (1,4-dioxan-2-yl)(4-((3-((2-((S)-1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone 64 (3.8 mg) in a yield of 14.33%.

[0816] MS m / z(ESI):488.9[M+1].

[0817] Example 65

[0818] (4-((4-(3-((2-((S)-1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(tetrahydro-2H-pyran-2-yl)methanone

[0819] (4-((4-(3-((2-((S)-1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(tetrahydro-2H-pyran-2-yl)methanone

[0820] first step

[0821] (tetrahydro-2H-pyran-2-yl)(4-((trimethylsilyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0822] (Tetrahydro-2H-pyran-2-yl)(4-((trimethylsilyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0823] 4-((Trimethylsilyl)ethynyl)-1,2,3,6-tetrahydropyridine 57a (50 mg, 278.81 μmol), tetrahydro-2H-pyran-2-carboxylic acid 65a (54.43 mg, 418.22 μmol, 47.74 μL, commercially available), O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (127.21 mg, 334.57 μmol) and N,N-diisopropylethylamine (108.10 mg, 836.43 μmol, 145.69 μL) were added sequentially to dichloromethane (1.95 mL) and stirred at room temperature for 4 hours. Mass spectrometry indicated that the reaction of the starting material was complete, and the system was concentrated to dryness under reduced pressure to give (tetrahydro-2H-pyran-2-yl)(4-((trimethylsilyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone 65b (60 mg), which was used directly in the next reaction.

[0824] MS m / z(ESI):292.0[M+1].

[0825] Step 2

[0826] (4-ethynyl-3,6-dihydropyridin-1(2H)-yl)(tetrahydro-2H-pyran-2-yl)methanone

[0827] (4-Ethynyl-3,6-dihydropyridin-1(2H)-yl)(tetrahydro-2H-pyran-2-yl)methanone

[0828] (Tetrahydro-2H-pyran-2-yl)(4-((trimethylsilyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone 65b (60 mg, 205.86 μmol) and potassium fluoride (59.80 mg, 1.03 mmol) were added sequentially to methanol (2 mL) and stirred at room temperature for 3 hours. The solution was concentrated to dryness under reduced pressure, dissolved in acetonitrile (3 mL), filtered, and the filtrate was concentrated to dryness under reduced pressure to afford (4-ethynyl-3,6-dihydropyridin-1(2H)-yl)(tetrahydro-2H-pyran-2-yl)methanone 65c (30 mg). The crude product was used directly in the next reaction. MS m / z (ESI): 220.1 [M+1].

[0829] Step 3

[0830] (tetrahydro-2H-pyran-2-yl)(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0831] (Tetrahydro-2H-pyran-2-yl)(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0832] 5-(4-iodophenyl)-3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazole 1i (30 mg, 59.46 μmol), (4-ethynyl-3,6-dihydropyridin-1(2H)-yl)(tetrahydro-2H-pyran-2-yl)methanone 65c (21.73 mg, 89.19 μmol), triethylenediamine (13.34 mg, 118.92 μmol) and allylpalladium(II) chloride dimer (4.34 mg, 11.89 μmol) were added sequentially to acetonitrile (2 mL), argon was replaced three times, and the mixture was stirred at room temperature for 12 hours. The system was concentrated to dryness under reduced pressure, and the residue was separated and purified by silica gel column chromatography (eluent: System A) to give (tetrahydro-2H-pyran-2-yl)(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone 65d (20 mg) in a yield of 58.94%.

[0833] MS m / z(ESI):570.9[M+1].

[0834] Step 4

[0835] (4-((4-(3-((2-((S)-1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(tetrahydro-2H-pyran-2-yl)methanone

[0836] (4-((4-(3-((2-((S)-1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(tetrahydro-2H-pyran-2-yl)methanone

[0837] (Tetrahydro-2H-pyran-2-yl)(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone 65d (20 mg, 35.05 μmol) and trifluoroacetic acid (0.5 mL) were added sequentially to dichloromethane (2 mL), and stirring was continued at room temperature for 1 hour. The system was concentrated to dryness under reduced pressure, and the residue was subjected to preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H2O, mobile phase B: CH3CN) to give (4-((4-(3-((2-((S)-1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(tetrahydro-2H-pyran-2-yl)methanone 65 (4.9 mg) in a yield of 22.44%.

[0838] MS m / z(ESI):486.9[M+1].

[0839] Example 66

[0840] (S)-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(tetrahydro-2H-pyran-4-yl)methanone

[0841] (S)-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(tetrahydro-2H-pyran-4-yl)methanone

[0842] first step

[0843] (tetrahydro-2H-pyran-4-yl)(4-((trimethylsilyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0844] (Tetrahydro-2H-pyran-4-yl)(4-((trimethylsilyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0845] 4-((Trimethylsilyl)ethynyl)-1,2,3,6-tetrahydropyridine 57a (50 mg, 278.81 μmol), tetrahydro-2H-pyran-4-carboxylic acid 66a (54.43 mg, 418.22 μmol), O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (127.21 mg, 334.57 μmol), and N,N-diisopropylethylamine (108.10 mg, 836.43 μmol, 145.69 μL) were added sequentially to dichloromethane (1.85 mL) and stirred at room temperature for 4 hours. Mass spectrometry indicated that the reaction of the starting materials was complete, and the system was concentrated to dryness under reduced pressure to obtain (tetrahydro-2H-pyran-4-yl)(4-((trimethylsilyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone 66b (50 mg), which was used directly in the next reaction.

[0846] MS m / z(ESI):292.0[M+1].

[0847] Step 2

[0848] (4-ethynyl-3,6-dihydropyridin-1(2H)-yl)(tetrahydro-2H-pyran-4-yl)methanone

[0849] (4-Ethynyl-3,6-dihydropyridin-1(2H)-yl)(tetrahydro-2H-pyran-4-yl)methanone

[0850] (Tetrahydro-2H-pyran-4-yl)(4-((trimethylsilyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone 66b (50 mg, 171.55 μmol) and potassium fluoride (49.84 mg, 857.75 μmol) were added sequentially to methanol (2 mL) and stirred at room temperature for 3 hours. The solution was concentrated to dryness under reduced pressure, dissolved in acetonitrile (3 mL), filtered, and the filtrate was concentrated to dryness under reduced pressure to afford (4-ethynyl-3,6-dihydropyridin-1(2H)-yl)(tetrahydro-2H-pyran-4-yl)methanone 66c (25 mg), which was used directly in the next reaction. MS m / z (ESI): 220.0 [M+1].

[0851] Step 3

[0852] (4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(tetrahydro-2H-pyran-4-yl)methanone

[0853] (4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(tetrahydro-2H-pyran-4-yl)methanone

[0854] 5-(4-iodophenyl)-3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazole 1i (30 mg, 59.46 μmol), (4-ethynyl-3,6-dihydropyridin-1(2H)-yl)(tetrahydro-2H-pyran-4-yl)methanone 66c (23.01 mg, 89.19 μmol), triethylenediamine (13.34 mg, 118.92 μmol) and allylpalladium(II) chloride dimer (4.34 mg, 11.89 μmol) were added sequentially to acetonitrile (2.0 mL), argon was replaced three times, and stirring was continued at room temperature for 3 hours. Ethyl acetate (30 mL) and water (15 mL) were added, the organic phase was separated, and the aqueous phase was extracted with ethyl acetate (30 mL×2). The combined organic phases were washed with saturated sodium chloride solution (30 mL×2), dried over anhydrous sodium sulfate, filtered, and concentrated to dryness under reduced pressure. The residue was separated and purified by silica gel column chromatography (eluent: System A) to give (4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(tetrahydro-2H-pyran-4-yl)methanone 66d (25 mg) in a yield of 73.68%.

[0855] MS m / z(ESI):571.0[M+1].

[0856] Step 4

[0857] (S)-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(tetrahydro-2H-pyran-4-yl)methanone

[0858] (S)-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(tetrahydro-2H-pyran-4-yl)methanone

[0859] (4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(tetrahydro-2H-pyran-4-yl)methanone 66d (20 mg, 35.05 μmol) and trifluoroacetic acid (0.5 mL) were added sequentially to dichloromethane (1 mL), and stirring was continued at room temperature for 1 hour. The system was concentrated to dryness under reduced pressure, and the residue was subjected to preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H2O, mobile phase B: CH3CN) to give (S)-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)(tetrahydro 2H-pyran-4-yl)methanone 66 (15.8 mg) in a yield of 71.39%.

[0860] MS m / z(ESI):486.9[M+1].

[0861] Example 67

[0862] (S)-1-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)-3-methoxypropan-1-one

[0863] (S)-1-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)-3-methoxypropan-1-one

[0864] first step

[0865] 3-methoxy-1-(4-((trimethylsilyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)propan-1-one

[0866] 3-Methoxy-1-(4-((trimethylsilyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)propan-1-one

[0867] 4-((Trimethylsilyl)ethynyl)-1,2,3,6-tetrahydropyridine 57a (50 mg, 278.81 μmol), 3-methoxypropionic acid 67a (43.54 mg, 418.22 μmol, 39.29 μL, commercially available), O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (127.21 mg, 334.57 μmol), and N,N-diisopropylethylamine (108.10 mg, 836.43 μmol, 145.69 μL) were added sequentially to dichloromethane (2 mL) and stirred at room temperature for 4 hours. Mass spectrometry indicated that the reaction of the starting material was complete, and the system was concentrated to dryness under reduced pressure to give 3-methoxy-1-(4-((trimethylsilyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)propan-1-one 67b (50 mg), which was used directly in the next reaction.

[0868] MS m / z(ESI):266.0[M+1].

[0869] Step 2

[0870] 1-(4-ethynyl-3,6-dihydropyridin-1(2H)-yl)-3-methoxypropan-1-one

[0871] 1-(4-Ethynyl-3,6-dihydropyridin-1(2H)-yl)-3-methoxypropan-1-one

[0872] 3-Methoxy-1-(4-((trimethylsilyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)propan-1-one 67b (50 mg, 188.38 μmol) and potassium fluoride (54.72 mg, 941.89 μmol) were added sequentially to methanol (2 mL) and stirred at room temperature for 3 hours. The solution was concentrated to dryness under reduced pressure, dissolved in acetonitrile (3 mL), filtered, and the filtrate was concentrated to dryness under reduced pressure to afford 1-(4-ethynyl-3,6-dihydropyridin-1(2H)-yl)-3-methoxypropan-1-one 67c (20 mg), which was used directly in the next reaction.

[0873] MS m / z(ESI):194.0[M+1].

[0874] Step 3

[0875] 3-methoxy-1-(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)propan-1-one

[0876] 3-Methoxy-1-(4-(4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)propan-1-one

[0877] 5-(4-iodophenyl)-3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazole 1i (30 mg, 59.46 μmol), 1-(4-ethynyl-3,6-dihydropyridin-1(2H)-yl)-3-methoxypropan-1-one 67c (20.28 mg, 89.19 μmol), triethylenediamine (13.34 mg, 118.92 μmol) and allylpalladium(II) chloride dimer (4.34 mg, 11.89 μmol) were added sequentially to acetonitrile (2.0 mL), argon was replaced three times, and stirring was continued at room temperature for 3 hours. Ethyl acetate (30 mL) and water (15 mL) were added, the organic phase was separated, and the aqueous phase was extracted with ethyl acetate (30 mL×2). The combined organic phases were washed with saturated sodium chloride solution (30 mL×2), dried over anhydrous sodium sulfate, filtered, and concentrated to dryness under reduced pressure. The residue was separated and purified by silica gel column chromatography (eluent: System A) to give 3-methoxy-1-(4-(4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)propan-1-one 67d (25 mg) in a yield of 77.20%.

[0878] MS m / z(ESI):545.0[M+1].

[0879] Step 4

[0880] (S)-1-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)-3-methoxypropan-1-one

[0881] (S)-1-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)-3-methoxypropan-1-one

[0882] 3-Methoxy-1-(4-(4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)propan-1-one 67d (20 mg, 36.72 μmol) and trifluoroacetic acid (0.5 mL) were added sequentially to dichloromethane (1 mL), and stirring was continued at room temperature for 1 hour. The system was concentrated to dryness under reduced pressure, and the residue was subjected to preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H 2 O, mobile phase B: CH 3 CN) to give (S)-1-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)-3-methoxypropan-1-one 67 (14.0 mg) in a yield of 64.70%.

[0883] MS m / z(ESI):461.0[M+1].

[0884] Example 68

[0885] (S)-1-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)-2-methoxyethan-1-one

[0886] (S)-1-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)-2-methoxyethan-1-one

[0887] first step

[0888] 2-methoxy-1-(4-((trimethylsilyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)ethan-1-one

[0889] 2-Methoxy-1-(4-((trimethylsilyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)ethan-1-one

[0890] 4-((Trimethylsilyl)ethynyl)-1,2,3,6-tetrahydropyridine 57a (50 mg, 278.81 μmol), 2-methoxyacetic acid 68a (37.67 mg, 418.22 μmol, commercially available), O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (127.21 mg, 334.57 μmol), and N,N-diisopropylethylamine (108.10 mg, 836.43 μmol, 145.69 μL) were added sequentially to dichloromethane (2 mL) and stirred at room temperature for 4 hours. Mass spectrometry indicated that the reaction of the starting material was complete, and the system was concentrated to dryness under reduced pressure to afford 2-methoxy-1-(4-((trimethylsilyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)ethan-1-one 68b (50 mg), which was used directly in the next reaction.

[0891] MS m / z(ESI):252.0[M+1].

[0892] Step 2

[0893] 1-(4-ethynyl-3,6-dihydropyridin-1(2H)-yl)-2-methoxyethan-1-one

[0894] 1-(4-Ethynyl-3,6-dihydropyridin-1(2H)-yl)-2-methoxyethan-1-one

[0895] 2-Methoxy-1-(4-((trimethylsilyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)ethan-1-one 68b (50 mg, 198.89 μmol) and potassium fluoride (57.78 mg, 994.45 μmol) were added sequentially to methanol (2 mL) and stirred at room temperature for 3 hours. The solution was concentrated to dryness under reduced pressure, dissolved in acetonitrile (3 mL), filtered, and the filtrate was concentrated to dryness under reduced pressure to afford 1-(4-ethynyl-3,6-dihydropyridin-1(2H)-yl)-2-methoxyethan-1-one 68c (20 mg), which was used directly in the next reaction.

[0896] MS m / z(ESI):180.0[M+1].

[0897] Step 3

[0898] 2-methoxy-1-(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)ethan-1-one

[0899] 2-Methoxy-1-(4-(4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)ethan-1-one

[0900] 5-(4-iodophenyl)-3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazole 1i (30 mg, 59.46 μmol), 1-(4-ethynyl-3,6-dihydropyridin-1(2H)-yl)-2-methoxyethan-1-one 68c (18.81 mg, 89.19 μmol), triethylenediamine (13.34 mg, 118.92 μmol) and allylpalladium(II) chloride dimer (4.34 mg, 11.89 μmol) were added sequentially to acetonitrile (2.0 mL), argon was replaced three times, and stirring was continued at room temperature for 3 hours. Ethyl acetate (30 mL) and water (15 mL) were added, the organic phase was separated, and the aqueous phase was extracted with ethyl acetate (30 mL×2). The combined organic phases were washed with saturated sodium chloride solution (30 mL×2), dried over anhydrous sodium sulfate, filtered, and concentrated to dryness under reduced pressure. The residue was separated and purified by silica gel column chromatography (eluent: System A) to give 2-methoxy-1-(4-(4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)ethan-1-one 68d (25 mg) in a yield of 79.24%.

[0901] MS m / z(ESI):531.0[M+1].

[0902] Step 4

[0903] (S)-1-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)-2-methoxyethan-1-one

[0904] (S)-1-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)-2-methoxyethan-1-one

[0905] 2-Methoxy-1-(4-(4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)ethan-1-one 68d (20 mg, 37.69 μmol) and trifluoroacetic acid (0.5 mL) were added sequentially to dichloromethane (1 mL), and stirring was continued at room temperature for 1 hour. The system was concentrated to dryness under reduced pressure, and the residue was subjected to preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H 2 O, mobile phase B: CH 3 CN) to give (S)-1-(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)-2-methoxyethan-1-one 68 (13.0 mg) in a yield of 58.89%.

[0906] MS m / z(ESI):446.9[M+1].

[0907] Example 69

[0908] (S)-(1-hydroxycyclopropyl)(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0909] (S)-(1-Hydroxycyclopropyl)(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0910] first step

[0911] (1-hydroxycyclopropyl)(4-((trimethylsilyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0912] (1-Hydroxycyclopropyl)(4-((trimethylsilyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0913] 4-((Trimethylsilyl)ethynyl)-1,2,3,6-tetrahydropyridine 57a (50 mg, 278.81 μmol), 1-hydroxycyclopropane-1-carboxylic acid 69a (42.69 mg, 418.22 μmol, commercially available), O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (127.21 mg, 334.57 μmol), and N,N-diisopropylethylamine (108.10 mg, 836.43 μmol, 145.69 μL) were added sequentially to dichloromethane (2 mL) and stirred at room temperature for 4 hours. Mass spectrometry indicated that the reaction of the starting material was complete, and the system was concentrated to dryness under reduced pressure to give (1-hydroxycyclopropyl)(4-((trimethylsilyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone 69b (50 mg), which was used directly in the next reaction.

[0914] MS m / z(ESI):263.9[M+1].

[0915] Step 2

[0916] (4-ethynyl-3,6-dihydropyridin-1(2H)-yl)(1-hydroxycyclopropyl)methanone

[0917] (4-Ethynyl-3,6-dihydropyridin-1(2H)-yl)(1-hydroxycyclopropyl)methanone

[0918] (1-Hydroxycyclopropyl)(4-((trimethylsilyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone 69b (50 mg, 189.82 μmol) and potassium fluoride (55.14 mg, 949.10 μmol) were added sequentially to methanol (2 mL) and stirred at room temperature for 3 hours. The system was concentrated to dryness under reduced pressure, dissolved in acetonitrile (3 mL), filtered, and the filtrate was concentrated to dryness under reduced pressure to afford (4-ethynyl-3,6-dihydropyridin-1(2H)-yl)(1-hydroxycyclopropyl)methanone 69c (30 mg), which was used directly in the next reaction.

[0919] MS m / z(ESI):191.9[M+1].

[0920] Step 3

[0921] (1-hydroxycyclopropyl)(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0922] (1-Hydroxycyclopropyl)(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0923] 5-(4-iodophenyl)-3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazole 1i (30 mg, 59.46 μmol), (4-ethynyl-3,6-dihydropyridin-1(2H)-yl)(1-hydroxycyclopropyl)methanone 69c (20.07 mg, 89.19 μmol), triethylenediamine (13.34 mg, 118.92 μmol) and allylpalladium(II) chloride dimer (4.34 mg, 11.89 μmol) were added sequentially to acetonitrile (2.0 mL), argon was replaced three times, and stirring was continued at room temperature for 3 hours. Ethyl acetate (30 mL) and water (15 mL) were added, and the layers were separated. The aqueous phase was extracted with ethyl acetate (30 mL × 2). The combined organic phases were washed with saturated sodium chloride solution (30 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated to dryness under reduced pressure. The residue was separated and purified by silica gel column chromatography (eluent: System A) to give (1-hydroxycyclopropyl)(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone 69d (25 mg) in a yield of 77.48%.

[0924] MS m / z(ESI):543.0[M+1].

[0925] Step 4

[0926] (S)-(1-hydroxycyclopropyl)(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0927] (S)-(1-Hydroxycyclopropyl)(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone

[0928] (1-Hydroxycyclopropyl)(4-((4-(3-((2-((1S)-1-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone 69d (20 mg, 36.86 μmol) and trifluoroacetic acid (0.5 mL) were added sequentially to dichloromethane (1 mL), and stirring was continued at room temperature for 1 hour. The system was concentrated to dryness under reduced pressure, and the residue was subjected to preparative liquid separation (separation column AKZONOBEL Kromasil; 250×21.2 mm; 5 μm, 20 mL / min; mobile phase A: 0.05% TFA+H 2 O, mobile phase B: CH 3 CN) to give (S)-(1-hydroxycyclopropyl)(4-((4-(3-((2-(1-hydroxyethyl)-1H-imidazol-1-yl)methyl)isoxazol-5-yl)phenyl)ethynyl)-3,6-dihydropyridin-1(2H)-yl)methanone 69 (2.2 mg) in a yield of 9.83%.

[0929] MS m / z(ESI):458.9[M+1].

[0930] Biological evaluation

[0931] Test Example 1: Determination of the Inhibition of LpxC Enzyme Activity by the Compounds of the Invention

[0932] The following method is used to determine the degree of inhibition of the compounds of the present invention on the enzymatic activity of recombinant Pseudomonas aeruginosa LpxC in vitro.

[0933] The experimental procedure is briefly summarized as follows: Test compounds were first dissolved in DMSO to a 10 mM stock solution. Reactions were performed in a 96-well microplate. First, 20 μL of recombinant Pseudomonas aeruginosa LpxC (purchased from Signalway Antibody, Catalog No. AP74647-2) was added to each well to a final concentration of 5 nM. Then, 5 μL of the test compound was added, and the compound was diluted fourfold to eight concentration points, ranging from 0.61 to 10,000 nM. Then, 5 μL of the LpxC substrate UDP-3-O-(R-3-hydroxydecanoyl)-GlcNAc (purchased from Biosynth Carbosynth, Catalog No. mu75071) was added to a final concentration of 10 μM. The cells were incubated at 25°C for 120 minutes. Then, 20 μL of 2.0 mg / mL fluorescamine (purchased from SigmaAldrich, product number F9015, solvent 1:1 dimethylformamide / acetonitrile) was added to the reaction system, mixed, and reacted for 10 minutes; finally, 50 μL of 200 mM sodium phosphate buffer (pH 8.0) was added to terminate the reaction and read using a microplate reader (BMG) with excitation and emission wavelengths of 390 and 495 nm, respectively. The percentage inhibition rate of the compound at each concentration was calculated by comparing the fluorescence intensity ratio with that of the control group (0.1% DMSO), and nonlinear regression analysis was performed using GraphPad Prism 5 software using the compound concentration logarithm-inhibition rate to obtain the compound's IC value. 50 The specific values ​​are shown in Table 1 below.

[0934] Table 1 Inhibition results of the compounds of the present invention on LpxC enzymatic activity

[0935] Conclusion: The IC values ​​of the preferred compounds of the present invention for inhibition of recombinant Pseudomonas aeruginosa LpxC enzyme activity are 50 <100nM, has a significant inhibitory effect on LpxC enzyme activity.

[0936] Test Example 2: Evaluation of the antibacterial activity of the compounds of the present invention

[0937] The in vitro minimum inhibitory concentration (MIC) determination was performed according to the CLSI guidelines using the broth microdilution method.

[0938] The experimental process is briefly described as follows: the test compound is dissolved in DMSO to prepare a 12.8 mg / mL stock solution, and then 11 two-fold dilutions of 100× high concentration working solutions are prepared using DMSO (the final concentration of the system is 64 μg / mL-0.06 μg / mL). The strains (K. Pneumoniae ATCC13883, K. Pneumoniae ATCC51504, and E. coli ATCC 25922) frozen in glycerol at -80°C are inoculated onto solid agar medium and placed in an incubator at 35°C for 18-24 hours to complete the strain preparation. Then, an appropriate amount of the solid plate culture is collected and resuspended in physiological saline, mixed, and the turbidity of the bacterial suspension is adjusted to an appropriate turbidity using a turbidimeter, containing approximately 1×10 8 cfu / mL bacteria, and then dilute the turbid bacterial suspension with test culture medium (type: CAMHIIB, purchased from BD Company) to a bacterial concentration of 5×10 5 cfu / mL to complete the inoculum preparation. 198 μL of the inoculum was inoculated into a 96-well plate, followed by the addition of 2 μL of a 100× high-concentration working solution of the compound. The 96-well plate was then incubated at 35°C for 18–24 hours. After incubation, the test plate was visually inspected. The lowest drug concentration that completely inhibited bacterial growth was the minimum inhibitory concentration (MIC) of the compound, as shown in Table 2.

[0939] Table 2 Antibacterial activity test results of the compounds of the present invention

[0940] Conclusion: The preferred compounds of the present invention have an in vitro minimum inhibitory degree of less than 50 μg / mL against K. pneumoniae (K. Pneumoniae ATCC13883, K. Pneumoniae ATCC51504) and Escherichia coli (E. coli ATCC 25922); they have good inhibitory effects on both K. pneumoniae and E. coli.

Claims

1. A compound represented by general formula (I) or its stereoisomers, tautomers or pharmaceutically acceptable salts: in: W is selected from O, S(O) r or N; Ring C is selected from a 5- to 10-membered spiro heterocyclic group or a monocyclic heterocyclic group, wherein the monocyclic heterocyclic group contains a double bond; X, Y, Z, Q are each independently selected from CR A or N atoms, and at most two atoms among X, Y, Z, and Q are N atoms at the same time; R A is selected from hydrogen, halogen, hydroxy, cyano, alkyl or alkoxy; wherein the alkyl or alkoxy is optionally further substituted by one or more substituents selected from halogen, hydroxy, cyano, alkyl or alkoxy; R 1 is selected from the group consisting of: absence, hydrogen, hydroxy, cyano, halogen, alkyl, alkoxy, cycloalkyl, heterocyclic, aryl, heteroaryl, -C(O)R 3 、-C(O)NR 6 R 7 or -S(O) r R 4 wherein the alkoxy, alkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl is optionally further substituted by one or more R B replaced by; R 3 、R 4 Each is independently selected from hydrogen atom, hydroxyl, alkyl, cycloalkyl, aryl, heteroaryl or heterocyclic group; wherein the alkyl, cycloalkyl, aryl, heteroaryl or heterocyclic group is optionally further substituted by one or more selected from hydroxyl, halogen, nitro, cyano, alkyl, alkoxy, haloalkyl, haloalkoxy, cycloalkyl, heterocyclic group, aryl, heteroaryl, =O, -C(O)R 8 、-C(O)OR 8 、-OC(O)R 8 、-NR 9 R 10 、-C(O)NR 9 R 10 、-SO2NR 9 R 10 or -NR 9 C(O)R 10 substituted by a substituent; R B are each independently selected from cyano, halogen, alkyl, hydroxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, -OR 5 、-C(O)R 5 、-C(O)OR 5 、-NHC(O)R 5 、-NHC(O)OR 5 、-NR 6 R 7 、-C(O)NR 6 R 7 、-CH2NHC(O)OR 5 、-CH2NR 6 R 7 or -S(O) r R 5 wherein the alkyl, cycloalkyl, heterocyclic, aryl or heteroaryl is optionally further substituted by one or more selected from hydroxy, halogen, nitro, cyano, alkyl, alkoxy, haloalkyl, haloalkoxy, cycloalkyl, heterocyclic, aryl, heteroaryl, =O, -C(O)R 8 、-C(O)OR 8 、-OC(O)R 8 、-NR 9 R 10 、-C(O)NR 9 R 10 、-SO2NR 9 R 10 or -NR 9 C(O)R 10 substituted by a substituent; Or, two R B Form a -C(=O)- with the same carbon atom to which it is attached; R 2 are the same or different, each independently selected from hydroxy, cyano, halogen, alkyl or alkoxy; wherein the alkyl or alkoxy is optionally further substituted with one or more substituents selected from halogen, hydroxy, cyano, alkyl or alkoxy; R 5 are independently selected from hydrogen atom, alkyl, cycloalkyl, heterocyclic, aryl or heteroaryl, wherein said alkyl, Cycloalkyl, heterocyclyl, aryl or heteroaryl may be further substituted by one or more radicals selected from hydroxy, halogen, nitro, cyano, alkyl, alkoxy, haloalkyl, haloalkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, =O, -C(O)R 8 、-C(O)OR 8 、-OC(O)R 8 、-NR 9 R 10 、-C(O)NR 9 R 10 、-SO2NR 9 R 10 or -NR 9 C(O)R 10 substituted by a substituent; R 6 and R 7 Each is independently selected from hydrogen, hydroxy, halogen, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl or heteroaryl, wherein the alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl or heteroaryl is optionally further substituted by one or more selected from hydroxy, halogen, nitro, cyano, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, =O, -C(O)R 8 、-C(O)OR 8 、-OC(O)R 8 、-NR 9 R 10 、-C(O)NR 9 R 10 、-SO2NR 9 R 10 or -NR 9 C(O)R 10 substituted by a substituent; Or, R 6 and R 7 Together with the atoms to which they are attached, they form a 4- to 8-membered heterocyclic group, wherein the 4- to 8-membered heterocyclic group contains one or more N, O or S(O)r, and the 4- to 8-membered heterocyclic group is optionally further substituted by one or more selected from hydroxy, halogen, nitro, cyano, alkyl, alkoxy, cycloalkyl, heterocyclic group, aryl, heteroaryl, =O, -C(O)R 8 、-C(O)OR 8 、-OC(O)R 8 、-NR 9 R 10 、-C(O)NR 9 R 10 、-SO2NR 9 R 10 or -NR 9 C(O)R 10 substituted by a substituent; R 8 、R 9 and R 10 Each is independently selected from a hydrogen atom, an alkyl group, an amino group, a cycloalkyl group, a heterocyclic group, an aryl group or a heteroaryl group, wherein the alkyl group, the cycloalkyl group, the heterocyclic group, the aryl group or the heteroaryl group is optionally further substituted with one or more substituents selected from a hydroxyl group, a halogen group, a nitro group, an amino group, a cyano group, an alkyl group, an alkoxy group, a cycloalkyl group, a heterocyclic group, an aryl group, a heteroaryl group, a carboxyl group or a carboxylate group; m is 0, 1 or 2; m is preferably 0; and r is independently 0, 1 or 2.

2. The compound according to claim 1 or its stereoisomer, tautomer or pharmaceutically acceptable salt, which is a compound or its stereoisomer, tautomer or pharmaceutically acceptable salt described by general formula (II): Ring C, R 1 、R 2 , W and m are as defined in claim 1.

3. The compound according to claim 1 or 2, or a stereoisomer, a tautomer or a pharmaceutically acceptable salt thereof, wherein W is N.

4. The compound according to claim 1 or 2, or a stereoisomer, tautomer, or a pharmaceutically acceptable salt thereof, wherein ring C is selected from:

5. The compound according to claim 1 or 2, or a stereoisomer, a tautomer, or a pharmaceutically acceptable salt thereof, wherein: R 1 Selected from hydrogen atom, hydroxyl group, alkyl group, cycloalkyl group, heterocyclic group, -C(O)R 3 、-C(O)NR 6 R 7 or -S(O) r R 4 wherein the alkyl or cycloalkyl group is optionally further substituted by one or more R B replaced by; R B Each is independently selected from cyano, hydroxy, heterocyclic, aryl, heteroaryl, -OR 5 、-C(O)OR 5 、-NR 6 R 7 、-C(O)NR 6 R 7 or -S(O) r R 5 wherein the heterocyclic or heteroaryl groups are optionally further substituted by one or more groups selected from hydroxy, cyano, alkoxy, haloalkyl, =O, -C(O)OR 8 or -C(O)NR 9 R 10 substituted by a substituent; Or, two R B Form a -C(=O)- with the same carbon atom to which it is attached; R 3 is selected from hydrogen atom, alkyl, cycloalkyl, heteroaryl or heterocyclic group; wherein the alkyl, cycloalkyl, heteroaryl or heterocyclic group is optionally further substituted by one or more selected from hydroxyl, alkoxy, cyano, heteroaryl, =O, -NR 9 R 10 、-C(O)OR 8 or -C(O)NR 9 R 10 substituted by a substituent; R 4 is a hydrogen atom or a hydroxyl group; R 5 are each independently selected from a hydrogen atom, an alkyl group or a hydroxyalkyl group; R 6 、R 7 are each independently selected from a hydrogen atom or an alkyl group, wherein the alkyl group is optionally substituted with one or more substituents selected from hydroxyl, cyano, halogen or alkoxy; R 8 、R 9 、R 10 Each is independently selected from a hydrogen atom or an alkyl group; the alkyl group is preferably a methyl group; r is 2.

6. The compound according to any one of claims 1 to 5, or its stereoisomer, tautomer, or pharmaceutically acceptable salt, wherein the compound is:

7. A pharmaceutical composition comprising an effective dose of the compound according to any one of claims 1 to 6, or its stereoisomers, tautomers, or pharmaceutically acceptable salts thereof, and a pharmaceutically acceptable carrier, excipient, or a combination thereof.

8. Use of the compound according to any one of claims 1 to 6, or its stereoisomer, tautomer, or pharmaceutically acceptable salt, or the pharmaceutical composition according to claim 7, in the preparation of an LPXC inhibitor.

9. Use of the compound according to any one of claims 1 to 6, or its stereoisomers, tautomers, or pharmaceutically acceptable salts thereof, or the pharmaceutical composition according to claim 7, in the preparation of a medicament for treating a disease mediated by LPXC, wherein the disease mediated by LPXC is selected from bacterial infections caused by Gram-negative bacteria.

10. The method according to claim 9, wherein the Gram-negative bacteria is selected from the group consisting of Escherichia coli, Pseudomonas aeruginosa, Proteus, Shigella dysenteriae, Klebsiella pneumoniae, Brucella, Salmonella typhi, Acinetobacter, Yersinia, Legionella pneumophila, Bordetella pertussis, Shigella, Pasteurella, Vibrio cholerae or Neisseria meningitidis.

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