ROCK inhibitors and uses thereof

Abstract

Disclosed herein are compounds of Formula (I) and pharmaceutically acceptable salts thereof useful as ROCK inhibitors. Also disclosed are pharmaceutical compositions comprising the compounds of Formula (I), and methods of using such compounds or compositions in the treatment of ROCK-related conditions (e.g., glaucoma).

Description

Invention Field

[0001] This disclosure generally relates to novel compounds that inhibit Rho-associated protein kinase (“ROCK”) and pharmaceutically acceptable salts thereof. This disclosure also relates to pharmaceutical compositions comprising said compounds as active ingredients, and the use of said compounds in the treatment of ROCK-related conditions, including glaucoma. Background Technology

[0002] Rho-associated protein kinase (ROCK) is a member of the serine-threonine protein kinase family. There are two isoforms of ROCK: ROCK1 and ROCK2. Both isoforms are activated by the GTP-binding form of Rho GTPases. ROCK plays a crucial role in many cellular processes, including smooth muscle cell contraction, cell proliferation, adhesion, and migration. Inhibition of ROCK activity has demonstrated potential therapeutic applicability across a wide range of pathological conditions.

[0003] Currently available ROCK inhibitor drugs include Eril from Asahi Kasei (used to treat cerebral vasospasm), Glanatec from Kowa (used to treat ocular hypertension and glaucoma), and Rhopressa from Aerie (used to lower elevated intraocular pressure (IOP) in patients with open-angle glaucoma or ocular hypertension).

[0004] Therefore, there is a need for novel compounds that inhibit rock as pharmacological tools, and these novel compounds have considerable value as drugs for treating rock-related conditions such as glaucoma. Summary of the Invention

[0005] This article discloses novel compounds capable of inhibiting rock. Therefore, the disclosed compounds can be used to treat rock-related diseases, such as glaucoma.

[0006] In one respect, this disclosure provides a compound of formula (I) or a pharmaceutically acceptable salt thereof:

[0007]

[0008] in

[0009] R 1 and R 2 Each is independently selected from the group consisting of: hydrogen, hydroxyl, halogen, cyano, amino, alkyl, alkenyl, ynyl, heteroalkyl, heteroalkenyl, and heteroynyl, wherein each of the alkyl, alkenyl, ynyl, heteroalkyl, heteroalkenyl, and heteroynyl is optionally substituted by one or more groups independently selected from: hydroxyl, halogen, cyano, or amino.

[0010] X, W, Z, and U are each independently N or C(R) 3 );

[0011] R 3 The group consisting of hydrogen, hydroxyl, halogen, cyano, amino, alkyl, alkenyl, ynyl, heteroalkyl, heteroalkenyl, and heteroynyl is selected from the group consisting of hydrogen, hydroxyl, halogen, cyano, or amino.

[0012] E is -N(R) 4 )C(=O)-、-C(=O)N(R 4 )-、-N(R 4 SO2- or -SO2N(R) 4 )-;

[0013] R 4 Selected from the following groups: hydrogen, alkyl, alkenyl, ynyl, heteroalkyl, heteroalkenyl, and heteroynyl;

[0014] Y is -Y 1 -Y 2 -(Y 3 ) n ,in

[0015] Y 1 Does not exist or -C(R) 5 )2-,

[0016] Each R 5 Independently selected from the following groups: hydrogen, -N(R) a 2. Alkyl, alkenyl, ynyl, heteroalkyl, heteroalkenyl, heteroynyl, cycloalkyl, heterocyclic, aryl, heteroaryl, and -alkyl-heterocyclic, wherein the alkyl, alkenyl, ynyl, heteroalkyl, heteroalkenyl, heteroynyl, cycloalkyl, heterocyclic, aryl, heteroaryl, and -alkyl-heterocyclic are optionally separated by one or more R 6 replace,

[0017] Each R 6 Independently selected from the group consisting of: hydroxyl, halogen, cyano, amino, alkyl, alkenyl, ynyl, heteroalkyl, heteroalkenyl, heteroynyl, and -alkyl-N(R) a )2;

[0018] Each R a Independently selected from hydrogen or alkyl;

[0019] Y 2 Selected from –(CH2) p -cycloalkyl-*、–(CH2) p -heterocyclic group-*, –(CH2)p -aryl-* or –(CH2) p -heteroaryl-*, each optionally substituted independently by one or more groups selected from: hydroxyl, halogen, cyano, amino, alkyl, alkenyl, ynyl, heteroalkyl, heteroalkenyl, or heteroynyl, and wherein p is 0, 1, or 2, and Y 2 The * end and Y 3 Connected;

[0020] Y 3 Does not exist or -Y 31 -Y 32 -Y 33 ;in

[0021] Y 31 Selected from non-existent, alkyl, alkenyl, or ynyl groups;

[0022] Y 32 Selected from: non-existent, -O-#, -OC(=O)-#, -C(=O)O-#, -P(=O)(R b )-#、-OC(=O)N(R b )-#、-N(R b )C(=O)-# or -C(=O)N(R b )-#, where each R b Independently selected from hydrogen or alkyl groups, and Y 32 #end and Y 33 connect;

[0023] Y 33 Choose from the following groups: hydrogen, hydroxyl, cyano, halogen, -N(R) c 2. Alkyl, alkenyl, ynyl, heteroalkyl, heteroalkenyl, heteroynyl, cycloalkyl, heterocyclic, aryl, and heteroaryl, wherein the alkyl, alkenyl, ynyl, heteroalkyl, heteroalkenyl, heteroynyl, cycloalkyl, heterocyclic, aryl, and heteroaryl groups are optionally separated by one or more R groups. 7 Replace, where each R 7 Independently selected from halogen, hydroxyl, amino, cyano, nitrooxy, alkyl, alkenyl, ynyl, heteroalkyl, heteroalkenyl, heteroynyl, or haloalkyl, and each R c Independently selected from hydrogen or alkyl; and

[0024] n is an integer from 1 to 5.

[0025] In another aspect, this disclosure provides a pharmaceutical composition comprising a compound of the disclosure or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

[0026] In another aspect, this disclosure provides a method for inhibiting ROCK activity in a subject in need, comprising administering to the subject an effective amount of a compound of this disclosure or a pharmaceutically acceptable salt thereof or a pharmaceutical composition of this disclosure.

[0027] In another aspect, this disclosure provides a method for treating ROCK-related conditions, comprising administering to a subject in need an effective amount of a compound of the present disclosure or a pharmaceutically acceptable salt thereof or a pharmaceutical composition of the present disclosure.

[0028] In another aspect, this disclosure provides the use of the compounds of this disclosure or pharmaceutically acceptable salts thereof or pharmaceutical compositions thereof in the manufacture of a medicament for treating ROCK-related conditions.

[0029] In another aspect, this disclosure provides a compound of the present disclosure or a pharmaceutically acceptable salt thereof or a pharmaceutical composition thereof for the treatment of ROCK-related conditions. Detailed Implementation

[0030] Certain embodiments of this disclosure will now be described in detail with reference to examples thereof, which are illustrated in the accompanying structures and formulas. While this disclosure will be described in conjunction with the enumerated embodiments, it should be understood that it is not intended to limit this disclosure to those embodiments. Rather, this disclosure is intended to cover all alternatives, modifications, and equivalents that may be included within the scope of this disclosure as defined in the claims. Those skilled in the art will recognize that many methods and materials similar to or equivalent to those described herein can be used to practice this disclosure. This disclosure is by no means limited to the methods and materials described. In the event that one or more of the incorporated references and similar materials (including, but not limited to, defined terms, usage of terms, described techniques, etc.) differ from or contradict this application, this disclosure shall prevail. All references, patents, and patent applications cited in this disclosure are hereby incorporated in their entirety by reference.

[0031] It should be understood that certain features of this disclosure described in the context of a single embodiment for clarity may also be provided in combination in a single embodiment. Conversely, various features of this disclosure described in the context of a single embodiment for brevity may also be provided individually or in any suitable sub-combination. It must be noted that, unless the context explicitly specifies otherwise, the singular forms “a / an” and “the” as used in the specification and appended claims include their plural forms. Thus, for example, reference to “a compound” includes multiple compounds.

[0032] definition

[0033] The definitions of specific functional groups and chemical terms are described in more detail below. For the purposes of this disclosure, chemical elements are identified according to the CAS version of the periodic table of elements in the 75th edition of the Handbook of Chemistry and Physics, and specific functional groups are generally defined as described therein. Furthermore, the general principles of organic chemistry, as well as specific functional parts and reactivity, are described in *Organic Chemistry*, Thomas Sorrell, University Science Books, Sosalito, 1999; *March's Advanced Organic Chemistry*, 5th ed., John Wiley & Sons, Inc., New York, 2001; *Comprehensive Organic Transformations*, Larock, VCH Publishers, Inc., New York, 1989; and *Some Modern Methods of Organic Synthesis*, 3rd ed., Cambridge University Press, Cambridge, 1987; the entire contents of each of these are incorporated herein by reference.

[0034] Linking substituents are described in several places in this disclosure. Where the structure explicitly requires a linking group, the Markush variable listed with respect to that group should be understood as the linking group. For example, if the structure requires a linking group and the Markush group definition of the variable lists "alkyl", then it should be understood that "alkyl" means linked alkylene.

[0035] When any variable (e.g., R) i When a group appears more than once in any component or formula of a compound, its definition for each occurrence is independent of its definition for each subsequent occurrence. Therefore, for example, if a group is shown as being surrounded by 0-2 R... i If partially substituted, then the group may optionally be replaced by up to two R groups. i Partial replacement, and R i Each time it appears, it is independently selected from R. i The definition of [the compound / compound] is as follows. Furthermore, combinations of substituents and / or variables are permitted, but only if such combinations produce stable compounds.

[0036] As used herein, for convenience, a dash “-” is used at the beginning or end of a chemical group to indicate the connection point of a substituent. For example, -OH is connected via an oxygen atom; chemical groups may or may not be described with one or more dashes without losing their ordinary meaning. A wavy line drawn through a line in the structure indicates the connection point of a group. Directionality is not indicated or implied by the order in which chemical groups are written or named unless required by chemistry or structure. As used herein, a solid line extending from the center of the ring indicates that the connection point of a substituent on the ring can be at any ring atom. When a substituent is listed without indicating the atom through which it bonds to the rest of the compound in the given formula, then the substituent can be bonded via any atom in the formula. Combinations of substituents and / or variables are permitted, but only if such combinations produce a stable compound.

[0037] Unless otherwise indicated herein, descriptions of ranges of values ​​are intended only as a shorthand for individually referring to each individual value falling within the range, and each individual value is incorporated into the description as if it were described individually herein. Unless otherwise stated, the ranges used herein include both limits of the range. For example, expressing "n is an integer between 1 and 6" and "n is an integer from 1 to 6" both mean "n is 1, 2, 3, 4, 5, or 6".

[0038] As used herein, the terms “compound provided herein” or “compound disclosed herein” or “compound disclosed herein” refer to compounds of formula (I) and the specific compounds disclosed herein.

[0039] As used in this article, the term "C" i-j "Indicates the range of carbon atoms, where i and j are integers, and the range includes the endpoints (i.e., i and j) and every integer point in between, where j is greater than i. For example, C..." 1-6 The term "C" indicates a range of one to six carbon atoms, including one, two, three, four, five, and six carbon atoms. In some embodiments, the term "C" is used... 1-12 "Indicates 1 to 12, especially 1 to 10, especially 1 to 8, especially 1 to 6, especially 1 to 5, especially 1 to 4, especially 1 to 3 or especially 1 to 2 carbon atoms."

[0040] Whether used as part of another term or independently, as used herein, the term "alkyl" refers to a saturated straight-chain or branched hydrocarbon radical that may optionally and independently be substituted by one or more substituents as described below. The term "C i-j"Alkyl" refers to an alkyl group having i to j carbon atoms. In some embodiments, the alkyl group contains 1 to 10 carbon atoms. In some embodiments, the alkyl group contains 1 to 9 carbon atoms. In some embodiments, the alkyl group contains 1 to 8 carbon atoms, 1 to 7 carbon atoms, 1 to 6 carbon atoms, 1 to 5 carbon atoms, 1 to 4 carbon atoms, 1 to 3 carbon atoms, or 1 to 2 carbon atoms. "C 1-10 Examples of "alkyl" include, but are not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, and decyl. "C" 1-6 Examples of "alkyl" are methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 2-pentyl, 3-pentyl, 2-methyl-2-butyl, 3-methyl-2-butyl, 3-methyl-1-butyl, 2-methyl-1-butyl, 1-hexyl, 2-hexyl, 3-hexyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 3-methyl-3-pentyl, 2-methyl-3-pentyl, 2,3-dimethyl-2-butyl, 3,3-dimethyl-2-butyl, etc.

[0041] Whether used as part of another term or independently, as used herein, the term "alkenyl" refers to a straight-chain or branched hydrocarbon radical having at least one carbon-carbon double bond, which may optionally be substituted independently by one or more substituents described herein, and includes radicals having "cis" and "trans" orientations, or alternatively "E" and "Z" orientations. In some embodiments, the alkenyl group contains 2 to 12 carbon atoms. In some embodiments, the alkenyl group contains 2 to 11 carbon atoms. In some embodiments, the alkenyl group contains 2 to 11 carbon atoms, 2 to 10 carbon atoms, 2 to 9 carbon atoms, 2 to 8 carbon atoms, 2 to 7 carbon atoms, 2 to 6 carbon atoms, 2 to 5 carbon atoms, 2 to 4 carbon atoms, 2 to 3 carbon atoms, and in some embodiments, the alkenyl group contains 2 carbon atoms. Examples of alkenyl groups include, but are not limited to, ethylenyl, allyl, butenyl, pentenyl, 1-methyl-2-but-1-yl, 5-hexenyl, etc.

[0042] Whether used as part of another term or independently, as used herein, the term "alkynyl" refers to a straight-chain or branched hydrocarbon radical having at least one carbon-carbon triple bond, which may optionally be independently substituted by one or more substituents as described herein. In some embodiments, the alkenyl group contains 2 to 12 carbon atoms. In some embodiments, the alkynyl group contains 2 to 11 carbon atoms. In some embodiments, the alkynyl group contains 2 to 11 carbon atoms, 2 to 10 carbon atoms, 2 to 9 carbon atoms, 2 to 8 carbon atoms, 2 to 7 carbon atoms, 2 to 6 carbon atoms, 2 to 5 carbon atoms, 2 to 4 carbon atoms, 2 to 3 carbon atoms, and in some embodiments, the alkynyl group contains 2 carbon atoms. Examples of alkynyl groups include, but are not limited to, ethynyl, 1-propynyl, 2-propynyl, etc.

[0043] As used herein, the term "amino" refers to the –NH2 group. Amino groups can also be substituted by one or more groups, such as alkyl, aryl, carbonyl, or other amino groups.

[0044] Whether used as part of another term or independently, as used herein, the term "aryl" refers to monocyclic and polycyclic ring systems having a total of 5 to 20 ring members, wherein at least one ring in the system is aromatic, and wherein each ring in the system contains 3 to 12 ring members. Examples of "aryl" include, but are not limited to, phenyl, biphenyl, naphthyl, anthracene, etc., which may have one or more substituents. As used herein, groups in which the aromatic ring is fused with one or more additional rings are also included within the scope of the term "aryl". In the case of polycyclic ring systems, only one ring needs to be aromatic (e.g., 2,3-dihydroindole), although all rings may be aromatic (e.g., quinoline). The second ring may also be fused or bridged. Examples of polycyclic aryl groups include, but are not limited to, benzofuranyl, indanyl, phthalimide, naphthimide, phenidyl, or tetrahydronaphthyl. The aryl group may be substituted at one or more ring positions with substituents as described above.

[0045] As used in this article, the term "cyano" refers to –CN.

[0046] Whether used as part of another term or independently, as used herein, the term "cycloalkyl" refers to a monovalent, non-aromatic, saturated or partially unsaturated monocyclic and polycyclic ring system in which all ring atoms are carbon, and the ring system contains at least three cyclic carbon atoms. In some embodiments, the cycloalkyl group may contain 3 to 12 cyclic carbon atoms, 3 to 10 cyclic carbon atoms, 3 to 9 cyclic carbon atoms, 3 to 8 cyclic carbon atoms, 3 to 7 cyclic carbon atoms, 3 to 6 cyclic carbon atoms, 3 to 5 cyclic carbon atoms, 4 to 12 cyclic carbon atoms, 4 to 10 cyclic carbon atoms, 4 to 9 cyclic carbon atoms, 4 to 8 cyclic carbon atoms, 4 to 7 cyclic carbon atoms, 4 to 6 cyclic carbon atoms, or 4 to 5 cyclic carbon atoms. The cycloalkyl group may be saturated or partially unsaturated. The cycloalkyl group may be substituted. In some embodiments, the cycloalkyl group may be a saturated cyclic alkyl group. In some embodiments, the cycloalkyl group may be a partially unsaturated cycloalkyl group containing at least one double or triple bond in its ring system. In some embodiments, the cycloalkyl group may be monocyclic or polycyclic. Fused, spiro, and bridging ring systems are also included within the scope of this definition. Examples of monocyclic cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopent-1-enyl, 1-cyclopent-2-enyl, 1-cyclopent-3-enyl, cyclohexyl, 1-cyclohex-1-enyl, 1-cyclohex-2-enyl, 1-cyclohex-3-enyl, cyclohexadienyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, cycloundecyl, and cyclododecyl. Examples of polycyclic cycloalkyl groups include, but are not limited to, adamantyl, norbornel, fluorenyl, spiropeptadienyl, spiro[3,6]decyl, bicyclo[1,1,1]pentenyl, bicyclo[2,2,1]heptenyl, etc.

[0047] As used herein, the term "halogen" refers to an atom selected from fluorine, chlorine, bromine, and iodine.

[0048] As used herein, the term “heteroatom” refers to nitrogen, oxygen, or sulfur, and includes any oxidized form of nitrogen or sulfur, as well as any quaternized form of basic nitrogen (including N-oxides).

[0049] As used herein, the term "heteroalkyl" refers to an alkyl group in which at least one carbon atom is replaced by a heteroatom selected from N, O, or S. A heteroalkyl group can be a carbon radical or a heteroatom radical (i.e., the heteroatom can be present in the middle or at the end of the radical) and can optionally be independently substituted by one or more substituents as described herein. The term "heteroalkyl" encompasses alkoxy and heteroalkoxy radicals.

[0050] As used herein, the term "heterenyl" refers to an alkenyl group in which at least one carbon atom is replaced by a heteroatom selected from N, O, or S. A heteroenyl group can be a carbon radical or a heteroatom radical (i.e., the heteroatom can be present in the middle or at the end of the radical) and can optionally be independently substituted by one or more substituents as described herein.

[0051] As used herein, the term "heterynyl" refers to an ynyl group in which at least one carbon atom is replaced by a heteroatom selected from N, O, or S. A heterynyl group can be a carbon radical or a heteroatom radical (i.e., the heteroatom can be present in the middle or at the end of the radical) and can optionally be independently substituted by one or more substituents as described herein.

[0052] Whether used as part of another term or independently, as used herein, the term "heteroaryl" refers to an aryl group having one or more heteroatoms in addition to a carbon atom. Heteroaryl groups can be monocyclic. Examples of monocyclic heteroaryl groups include, but are not limited to, thiophene, furanyl, pyrrole, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridinyl, pyridazinyl, pyrazinyl, indoleazinyl, purine, naphridinyl, benzofuranyl, and pteridinyl. Heteroaryl groups also include polycyclic groups in which the heteroaryl ring is fused with one or more aryl, cycloaliphatic, or heterocyclic rings, wherein the linking radical or linking point is located on the heteroaryl ring. Examples of polycyclic heteroaryl groups include, but are not limited to, indolyl, isoindolyl, benzothiophene, benzofuranyl, benzo[1,3]dioxacyclopentenyl, dibenzofuranyl, indazoleyl, benzimidazolyl, benzothiazolyl, quinolinyl, isoquinolinyl, dihydroquinolinyl, dihydroisoquinolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, terolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H-quinazinyl, carbazolyl, acridineyl, phenazinyl, phenothiazinyl, phenotoxazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, etc.

[0053] As used herein, the term "heterocyclic group" refers to a carbocyclic group in which one or more ring atoms are independently selected from heteroatoms of oxygen, sulfur, nitrogen, phosphorus, etc., and the remaining ring atoms are saturated or partially unsaturated carbon, wherein one or more ring atoms may optionally be independently substituted by one or more substituents. In some embodiments, the heterocyclic group is a saturated heterocyclic group. In some embodiments, the heterocyclic group is a partially unsaturated heterocyclic group having one or more double bonds in its ring system. In some embodiments, the heterocyclic group may contain any oxidized form of carbon, nitrogen, or sulfur and any quaternized form of basic nitrogen. "Heterocyclic group" also includes a radical in which the heterocyclic radical is fused with a saturated, partially unsaturated, or fully unsaturated (i.e., aromatic) carbocyclic or heterocyclic ring. Where possible, the heterocyclic radical may be carbon-linked or nitrogen-linked. In some embodiments, the heterocycle is carbon-linked. In some embodiments, the heterocycle is nitrogen-linked. For example, a group derived from pyrrole may be pyrrole-1-yl (nitrogen-linked) or pyrrole-3-yl (carbon-linked). Furthermore, the group derived from imidazole can be imidazole-1-yl (nitrogen-linked) or imidazole-3-yl (carbon-linked).

[0054] In some embodiments, the term "3- to 12-membered heterocyclic group" refers to a 3- to 12-membered saturated or partially unsaturated monocyclic or polycyclic heterocyclic ring system having 1 to 3 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Fused, spiro, and bridging ring systems are also included within the scope of this definition. Examples of monocyclic heterocyclic groups include, but are not limited to, oxoheterocyclic butyl, 1,1-dioxothiocyclic butylpyrrolidinyl, tetrahydrofuranyl, tetrahydrothiophenyl, pyrrolidinyl, furanyl, thiophenyl, pyrazolyl, imidazolyl, triazolyl, oxazolyl, thiazolyl, piperidyl, piperazinyl, piperidinyl, morpholinyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, triazinyl, pyridinoneyl, pyridinoneyl, pyridinoneyl, pyridinoneyl, pyrrolidinyl, triazinoneyl, etc. Examples of fused heterocyclic groups include, but are not limited to, phenyl fused rings or pyridyl fused rings, such as quinolinyl, isoquinolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, quinoxalinyl, quinazinyl, quinazolinyl, azaindolazinyl, pteridinyl, chromenyl, isochromenyl, indolyl, isoindolyl, indolazinyl, indazole, purine, benzofuranyl, isobenzofuranyl, benzimidazolyl, benzothiopheneyl, benzothiazolyl, carbazole, phenazinyl, phenthiazolyl, phenanthidyl, hexahydro-1H-pyrrolazinyl, imidazo[1,2-a]pyridinyl, [1,2,4]triazolo[4,3-a]pyridinyl, [1,2,3]triazolo[4,3-a]pyridinyl, etc. Examples of spiroheterocyclic groups include, but are not limited to, spiropyranyl, spiroxazinyl, etc. Examples of bridging heterocyclic groups include, but are not limited to, morphanyl, hexamethylenetetramine, 3-azabicyclo[3.1.0]hexane, 8-azabicyclo[3.2.1]octane, 1-azabicyclo[2.2.2]octane, and 1,4-diazabicyclo[2.2.2]octane (DABCO).

[0055] As used in this article, the term "hydroxyl" refers to –OH.

[0056] As used herein, the term "partially unsaturated" refers to a radical that includes at least one double or triple bond. The term "partially unsaturated" is intended to cover rings with multiple unsaturated sites, but is not intended to include aromatic (i.e., completely unsaturated) portions.

[0057] As used herein, the terms “optional” or “optionally” mean that the event or condition described below may or may not occur, and the description includes examples of the event or condition occurring and examples of the event or condition not occurring. As used herein, regardless of whether the term “optionally” is preceding it, the term “substituted” means that one or more hydrogens of the specified moiety are replaced by a suitable substituent. It should be understood that “substituted” or “replaced by” includes the following implicit limitations: the substitution is consistent with the permissible valence state of the substituted atom, and the substitution produces a stable or chemically viable compound, such as a compound that does not spontaneously transform by rearrangement, cyclization, elimination, etc. Unless otherwise indicated, the “optionally substituted” group may have a suitable substituent at each substituted position of the group, and when more than one position in any given structure can be replaced by more than one substituent selected from the specified group, the substituents at each position may be the same or different. Substituents may include, but are not limited to, alkyl, alkenyl, alkynyl, alkoxy, acyl, amino, amide, amidine, aryl, azide, carbamoyl, carboxyl, carboxyl ester, cyano, guanidinyl, halogen, haloalkyl, heteroalkyl, heteroaryl, heterocyclic, hydroxyl, hydrazine, imino, oxy, nitro, alkylsulfinyl, sulfonic acid, alkylsulfonyl, thiocyanate, thiol, thion, or combinations thereof. Those skilled in the art will understand that the substituent itself may be substituted, if appropriate. Unless specifically stated as “unsubstituted,” references to the chemical part herein should be understood to include substituted variants. For example, references to the “aryl” group or part thereof implicitly include both substituted and unsubstituted variants.

[0058] As used herein, regardless of whether the term "optionally" is preceding it, the term "substituted" means that one or more hydrogens of the specified moiety are replaced by a suitable substituent. It should be understood that "substituted" or "replaced by" includes the implied limitation that the substitution is consistent with the permissible valence state of the substituted atom and that the substitution produces a stable or chemically viable compound, such as a compound that does not spontaneously transform through rearrangement, cyclization, elimination, etc. Unless otherwise indicated, a "optionally substituted" group may have a suitable substituent at each substituted position of the group, and when more than one position in any given structure can be substituted by more than one substituent selected from the specified group, the substituents at each position may be the same or different. Those skilled in the art will understand that the substituent itself may be substituted, if appropriate. Unless specifically stated as "unsubstituted," references to the chemical portion herein should be understood to include substituted variants. For example, a reference to an "aryl" group or part implicitly includes both substituted and unsubstituted variants.

[0059] The symbols “R” and “S” denote the configuration of the substituents surrounding the chiral carbon atom. As described herein, the isomer descriptors “R” and “S” are used to indicate the atomic configuration relative to the core molecule and are intended to be used as defined in the literature (IUPAC Recommendations 1996, Pure and Applied Chemistry, 68:2193-2222 (1996)).

[0060] compound

[0061] In one respect, this disclosure provides compounds of formula (I) or pharmaceutically acceptable salts thereof:

[0062]

[0063] in

[0064] R 1 and R 2 Each is independently selected from the group consisting of: hydrogen, hydroxyl, halogen, cyano, amino, alkyl, alkenyl, ynyl, heteroalkyl, heteroalkenyl, and heteroynyl, wherein each of the alkyl, alkenyl, ynyl, heteroalkyl, heteroalkenyl, and heteroynyl is optionally substituted by one or more groups independently selected from: hydroxyl, halogen, cyano, or amino.

[0065] X, W, Z, and U are each independently N or C(R) 3 );

[0066] R 3 The group consisting of hydrogen, hydroxyl, halogen, cyano, amino, alkyl, alkenyl, ynyl, heteroalkyl, heteroalkenyl, and heteroynyl is selected from the group consisting of hydrogen, hydroxyl, halogen, cyano, or amino.

[0067] E is -N(R) 4 )C(=O)-、-C(=O)N(R 4 )-、-N(R 4 SO2- or -SO2N(R) 4 )-;

[0068] R 4 Selected from the following groups: hydrogen, alkyl, alkenyl, ynyl, heteroalkyl, heteroalkenyl, and heteroynyl;

[0069] Y is -Y 1 -Y 2 -(Y 3 ) n ,in

[0070] Y 1 Does not exist or -C(R) 5 )2-,

[0071] Each R 5 Independently selected from the following groups: hydrogen, -N(R) a 2. Alkyl, alkenyl, ynyl, heteroalkyl, heteroalkenyl, heteroynyl, cycloalkyl, heterocyclic, aryl, heteroaryl, and -alkyl-heterocyclic, wherein the alkyl, alkenyl, ynyl, heteroalkyl, heteroalkenyl, heteroynyl, cycloalkyl, heterocyclic, aryl, heteroaryl, and -alkyl-heterocyclic are optionally separated by one or more R 6 replace,

[0072] Each R 6 Independently selected from the group consisting of: hydroxyl, halogen, cyano, amino, alkyl, alkenyl, ynyl, heteroalkyl, heteroalkenyl, heteroynyl, and -alkyl-N(R) a )2;

[0073] Each R a Independently selected from hydrogen or alkyl;

[0074] Y 2 Selected from –(CH2) p -cycloalkyl-*、–(CH2) p -heterocyclic group-*, –(CH2) p -aryl-* or –(CH2) p -heteroaryl-*, each optionally substituted independently by one or more groups selected from: hydroxyl, halogen, cyano, amino, alkyl, alkenyl, ynyl, heteroalkyl, heteroalkenyl, or heteroynyl, and wherein p is 0, 1, or 2, and Y 2 The * end and Y 3 Connected;

[0075] Y 3 Does not exist or -Y 31 -Y 32 -Y 33 ;in

[0076] Y 31 Selected from non-existent, alkyl, alkenyl, or ynyl groups;

[0077] Y 32 Selected from: non-existent, -O-#, -OC(=O)-#, -C(=O)O-#, -P(=O)(R b )-#、-OC(=O)N(R b )-#、-N(R b )C(=O)-# or -C(=O)N(R b )-#, where each Rb Independently selected from hydrogen or alkyl groups, and Y 32 #end and Y 33 connect;

[0078] Y 33 Choose from the following groups: hydrogen, hydroxyl, cyano, halogen, -N(R) c 2. Alkyl, alkenyl, ynyl, heteroalkyl, heteroalkenyl, heteroynyl, cycloalkyl, heterocyclic, aryl, and heteroaryl, wherein the alkyl, alkenyl, ynyl, heteroalkyl, heteroalkenyl, heteroynyl, cycloalkyl, heterocyclic, aryl, and heteroaryl groups are optionally separated by one or more R groups. 7 Replace, where each R 7 Independently selected from halogen, hydroxyl, amino, cyano, nitrooxy, alkyl, alkenyl, ynyl, heteroalkyl, heteroalkenyl, heteroynyl, or haloalkyl, and each R c Independently selected from hydrogen or alkyl; and

[0079] n is an integer from 1 to 5.

[0080] In some implementations, R 1 and R 2 Each is independently hydrogen, halogen, hydrogen, cyano, amino, or an alkyl group optionally substituted with one or more halogens.

[0081] In some implementations, R 1 It is hydrogen or alkyl, and R 2 It is hydrogen, halogen, or alkyl. In some embodiments, R 1 Is it hydrogen or C? 1-6 Alkyl, and R 2 Is it hydrogen, halogen, or C? 1-6 alkyl.

[0082] In some implementations, R 1 and R 2 They're all hydrogen.

[0083] In some implementations, R 1 and R 2 Both are alkyl groups. In some embodiments, R 1 It is C 1-6 Alkyl, C 1-5 Alkyl, C 1-4 Alkyl, C 1-3 Alkyl or C 1-2 Alkyl, and R 2 It is C 1-6 Alkyl, C 1-5 Alkyl, C 1-4 Alkyl, C 1-3 Alkyl or C 1-2 Alkyl group. In some embodiments, R1 It is methyl, and R 2 It is a methyl group.

[0084] In some implementations, R 1 It is hydrogen, and R 2 It is a halogen. In some embodiments, R 1 It is hydrogen, and R 2 It's fluorine.

[0085] In some implementations, R 1 It is hydrogen, and R 2 It is an alkyl group. In some embodiments, R 1 It is hydrogen, and R 2 It is C 1-6 Alkyl, C 1-5 Alkyl, C 1-4 Alkyl, C 1-3 Alkyl or C 1-2 Alkyl group. In some embodiments, R 1 It is hydrogen, and R 2 It is a methyl group.

[0086] In some implementations, X, W, U, and Z are C(R) 3 In some implementations, W, U, and Z are C(R). 3 ), and R 3 It is hydrogen.

[0087] In some implementations, X and U are C(R) 3 ), and W and Z are N. In some implementations, X and U are C(R) 3 ), where R 3 It is hydrogen, and W and Z are N.

[0088] In some implementations, E is -N(R) 4 )C(=O)- or -C(=O)N(R 4 In some implementations, E is -N(R) 4 )C(=O)- or -C(=O)N(R 4 )-, and R 4 It is hydrogen.

[0089] In some implementations, Y 1 It does not exist.

[0090] In some implementations, Y 1 It is -C(R) 5 )2-.

[0091] In some implementations, Y 1 It is -C(R) 5 )2-, an R5 It's hydrogen, another R 5 Selected from -N(R) a 2. Alkyl, cycloalkyl, heterocyclic, and –alkyl-heterocyclic groups, wherein the alkyl, cycloalkyl, heterocyclic, and –alkyl-heterocyclic groups are optionally composed of one or more R groups. 6 replace.

[0092] In some implementations, Y 1 It is -C(R) 5 )2-, an R 5 It's hydrogen, another R 5 Selected from -N(R) a 2. C 1-6 Alkyl, C 3-12 Cycloalkyl, 5- to 12-membered heterocyclic or –(C 1-6 alkyl)-(5 to 12-membered heterocyclic group), wherein the C 1-6 Alkyl, C 3-12 Cycloalkyl, 5- to 12-membered heterocyclic groups and –(C 1-6 Alkyl group (5 to 12-membered heterocyclic group) optionally surrounded by one or more R groups 6 replace.

[0093] In some implementations, Y 1 It is -C(R) 5 )2-, an R 5 It is an alkyl group, another R 5 Selected from -N(R) a 2. Alkyl, cycloalkyl, or heterocyclic group, wherein the alkyl, cycloalkyl, or heterocyclic group is optionally surrounded by one or more R groups. 6 replace.

[0094] In some implementations, Y 1 It is -C(R) 5 )2-, an R 5 It is C 1-6 Alkyl, another R 5 Selected from -N(R) a 2. C 1-6 Alkyl, C 3-12 cycloalkyl or 5- to 12-membered heterocyclic groups, wherein the C 1-6 Alkyl, C 3-12 Cycloalkyl or 5- to 12-membered heterocyclic groups are optionally surrounded by one or more R 6 replace.

[0095] In some implementations, R 6 Selected from amino, alkyl, or -alkyl-N(R) a 2. In some implementations, R 6 Selected from amino, C 1-6 Alkyl or –(C1-6 alkyl)-N(R a )2.

[0096] In some implementations, Y 1 Choose from the following groups:

[0097]

[0098] In some implementations, Y 2 It is –(CH2) p -cycloalkyl-*.

[0099] In some implementations, Y 2 It is –(CH2) p -(C 3-10 (cycloalkyl)-*, and p is 0 or 1.

[0100] In some implementations, Y 2 It is cyclopentyl or cyclohexyl.

[0101] In some implementations, Y 2 It is –(CH2) p -Heterocyclic group-*.

[0102] In some implementations, Y 2 It is –(CH2) p -(5 to 12-membered heterocyclic group)-*, and p is 0 or 1.

[0103] In some implementations, Y 2 It is tetrahydrofuranyl, pyrrolidinyl, or tetrahydropyranyl.

[0104] In some implementations, Y 2 It is –(CH2) p -Aryl-*.

[0105] In some implementations, Y 2 It is –(CH2) p -(C 5-12 Aryl)-*, and p is 0 or 1.

[0106] In some implementations, Y 2 It is phenyl or -CH2-phenyl-*.

[0107] In some implementations, Y 2 It is –(CH2) p -Mixingal aryl-*.

[0108] In some implementations, Y 2 It is –(CH2) p-(5 to 12 heteroaryl)-*, and p is 0 or 1.

[0109] In some implementations, Y 2 It is pyridyl, tetrahydroisoquinolinyl, -CH2-imidazolyl-*, or -CH2-indolyl-*.

[0110] In some implementations, Y 31 It is absent or alkyl. In some embodiments, Y 31 Does not exist or C 1-6 Alkyl, C 1-5 Alkyl, C 1-4 Alkyl, C 1-3 Alkyl or C 1-2 alkyl.

[0111] In some implementations, Y 31 It does not exist, and Y 32 Selected from: non-existent, -O-#, -C(=O)O-#, -P(=O)(R b )-#、-N(R b )C(=O)-# or -C(=O)N(R b )-#.

[0112] In some implementations, Y 31 It does not exist, Y 32 Selected from: non-existent, -O-#, -C(=O)O-#, -P(=O)(R b )-#、-N(R b )C(=O)-# or -C(=O)N(R b )-#, and Y 33 Selected from -N(R) c 2. Alkyl or aryl, wherein the alkyl and aryl groups are optionally separated by one or more R groups. 7 replace.

[0113] In some implementations, Y 31 It does not exist, Y 32 Selected from: non-existent, -O-#, -C(=O)O-#, -P(=O)(R b )-#、-N(R b )C(=O)-# or -C(=O)N(R b )-#, and Y 33 Selected from -N(R) c 2. C 1-6 Alkyl or C 5-12 Aryl, wherein the C 1-6 Alkyl and C 5-12 Aryl group is optionally bounded by one or more R 7 replace.

[0114] In some implementations, Y 31 It does not exist, Y 32 Selected from: non-existent, -O-#, -C(=O)O-#, -P(=O)(R b )-#、-N(R b )C(=O)-# or -C(=O)N(R b )-#, and Y 33 Selected from -N(R) c 2. C 1-6 Alkyl or C 5-12 Aryl, wherein the C 1-6 Alkyl and C 5-12 The aryl group is optionally and independently selected from one or more of the following R groups. 7 Substitution: halogen, hydroxyl, amino, cyano, nitrooxy or alkyl.

[0115] In some implementations, Y 31 It does not exist, Y 32 Selected from: non-existent, -O-#, -C(=O)O-#, -P(=O)(R b )-#、-N(R b )C(=O)-# or -C(=O)N(R b )-#, and Y 33 Selected from hydrogen, hydroxyl, cyano, halogen, -NH2, methyl, -CH2CH2OCH3.

[0116] In some implementations, Y 31 It is an alkyl group, and Y 32 Selected from: non-existent, -O-#, -OC(=O)-#, -OC(=O)N(R) b -# or -N(R) b )C(=O)-#.

[0117] In some implementations, Y 31 It is C 1-6 Alkyl, C 1-5 Alkyl, C 1-4 Alkyl, C 1-3 Alkyl or C 1-2 Alkyl, and Y 32 Selected from: non-existent, -O-#, -OC(=O)-#, -OC(=O)N(R) b -# or -N(R) b )C(=O)-#.

[0118] In some implementations, Y 31 It is C 1-6 Alkyl, C 1-5 Alkyl, C 1-4 Alkyl, C1-3 Alkyl or C 1-2 Alkyl, Y 32 Selected from: non-existent, -O-#, -OC(=O)-#, -OC(=O)N(R) b -# or -N(R) b )C(=O)-#, and Y 33 Selected from -N(R) c 2. Alkyl or aryl, wherein the alkyl and aryl groups are optionally separated by one or more R groups. 7 replace.

[0119] In some implementations, Y 31 It is C 1-6 Alkyl, C 1-5 Alkyl, C 1-4 Alkyl, C 1-3 Alkyl or C 1-2 Alkyl, Y 32 Selected from: non-existent, -O-#, -OC(=O)-#, -OC(=O)N(R) b -# or -N(R) b )C(=O)-#, and Y 33 The group is selected from –NH2, -N(CH3)2, methyl, dimethylphenyl, or nitrooxypentyl, wherein the alkyl and aryl groups are optionally separated by one or more R groups. 7 replace.

[0120] In some implementations, Y 31 It is C 1-6 Alkyl, C 1-5 Alkyl, C 1-4 Alkyl, C 1-3 Alkyl or C 1-2 Alkyl, Y 32 Selected from: non-existent, -O-#, -OC(=O)-#, -OC(=O)N(R) b -# or -N(R) b )C(=O)-#, and Y 33 Selected from –NH2, -N(CH3)2, methyl, dimethylphenyl or nitrooxypentyl.

[0121] In some implementations, n is 1 or 2.

[0122] Exemplary compounds of formula (I) are stated in Table 1 below.

[0123] Table 1. Exemplary compounds of formula (I)

[0124]

[0125]

[0126]

[0127]

[0128]

[0129]

[0130]

[0131]

[0132]

[0133]

[0134]

[0135]

[0136]

[0137] The compounds described herein are illustrated with reference to general formulas and specific compounds. Furthermore, the compounds disclosed herein may exist in many different forms or derivatives, including but not limited to prodrugs, active metabolic derivatives (active metabolites), solvates, pharmaceutically acceptable salts, or isotopic derivatives, all of which are within the scope of this disclosure.

[0138] As used herein, the term "prodrug" refers to a compound or a pharmaceutically acceptable salt thereof that, when metabolized under physiological conditions or when converted by solvent degradation, yields the desired active compound. Prodrugs include, but are not limited to, esters, amides, carbamates, carbonates, acylureas, solvates, or hydrates of the active compound. Typically, prodrugs are inactive or less active than the active compound, but may provide one or more of the advantageous treatment, administration, and / or metabolic properties. For example, some prodrugs are esters of the active compound; during metabolic degradation, the ester group is cleaved to produce the active drug. Furthermore, some prodrugs are activated by enzymes to produce the active compound, or to produce compounds that yield the active compound after further chemical reactions. Prodrugs can develop from their prodrug form to their active form in a single step, or may have one or more intermediate forms that may or may not be active. The preparation and use of prodrugs are discussed in T. Higuchi and V. Stella, “Pro-drugs as Novel Delivery Systems,” Proceedings of the ACS Symposium Series, Vol. 14, Bioreversible Carriers in Drug Design, edited by Edward B. Roche, American Pharmaceutical Association and Pergamon Press, 1987; and in Prodrugs: Challenges and Rewards, edited by V. Stella, R. Borchardt, M. Hageman, R. Oliyai, H. Maag, and J. Tilley, Springer-Verlag New York, 2007. All of these references are incorporated herein by reference in their entirety.

[0139] As used herein, the term "metabolite," such as an active metabolite, overlaps with the prodrugs described above. Therefore, such metabolites are pharmacologically active compounds or compounds further metabolized into pharmacologically active compounds, which are derivatives produced by metabolic processes within the subject's body. For example, such metabolites can be generated by oxidation, reduction, hydrolysis, amidation, deamidation, esterification, deesterification, enzymatic cleavage, etc., of the administered compound or salt or prodrug. Active metabolites are such pharmacologically active derivatives. For prodrugs, the prodrug compound is generally inactive or less active than the metabolite. For active metabolites, the parent compound can be an active compound or an inactive prodrug.

[0140] Prodrugs and active metabolites can be identified using conventional techniques known in the art. See, for example, Bertolini et al., 1997, *Journal of Medicinal Chemistry* 40: 2011-2016; Shan et al., *Journal of Pharmacy* 86: 756-757; Bagshawe, 1995, *DrugDevRes* 34: 220-230; Wermuth, ibid.

[0141] As used herein, the term "pharmaceutically acceptable" indicates that the substance or composition is chemically and / or toxicologically compatible with the other components constituting the formulation and / or with the subjects treated therein.

[0142] As used herein, unless otherwise indicated, the term "pharmaceutically acceptable salt" includes salts that retain the bioavailability of the specified compound as a free acid and base and are not biologically or otherwise undesirable. Considered pharmaceutically acceptable salt forms include, but are not limited to, monosalts, disalts, trisalts, tetrasalts, etc. Pharmaceutically acceptable salts are non-toxic at the amount and concentration at which they are administered. The preparation of such salts can facilitate pharmacological use by altering the physical properties of the compound without impairing its physiological effects. Useful alterations to physical properties include lowering the melting point to facilitate transmucosal administration and increasing solubility to facilitate administration of higher drug concentrations.

[0143] Pharmaceutically acceptable salts include acid addition salts, such as those containing sulfates, chlorides, hydrochlorides, fumarates, maleates, phosphates, aminosulfonates, acetates, citrates, lactates, tartrates, methanesulfonates, ethanesulfonates, benzenesulfonates, p-toluenesulfonates, cyclohexylaminosulfonates, and quinates. Pharmaceutically acceptable salts can be obtained from acids such as hydrochloric acid, maleic acid, sulfuric acid, phosphoric acid, aminosulfonic acid, acetic acid, citric acid, lactic acid, tartaric acid, malonic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, cyclohexylaminosulfonic acid, fumaric acid, and quinic acid.

[0144] Pharmaceutically acceptable salts also include base addition salts, such as those containing benzathine, chloroprocaine, choline, diethanolamine, ethanolamine, tert-butylamine, ethylenediamine, meglumine, procaine, aluminum, calcium, lithium, magnesium, potassium, sodium, ammonium, alkylamines, and zinc, when acidic functional groups such as carboxylic acid or phenol are present. See, for example, *Remington's Pharmaceutical Sciences*, 19th edition, Mack Publishing Co., Easton, PA, Vol. 2, p. 1457, 1995; "Handbook of Pharmaceutical Salts: Properties, Selection, and Use," Stahl and Wermuth, Wiley-VCH, Weinheim, Germany, 2002. These salts can be prepared using appropriate corresponding bases.

[0145] Pharmaceutically acceptable salts can be prepared using standard techniques. For example, the free base form of a compound can be dissolved in a suitable solvent, such as an aqueous solution or a water-alcohol solution containing a suitable acid, and then separated by evaporation of the solution. Therefore, if a particular compound is a base, the desired pharmaceutically acceptable salt can be prepared by any suitable method available in the art, for example, by treating the free base with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc., or with organic acids such as acetic acid, maleic acid, succinic acid, mandelic acid, fumaric acid, malonic acid, pyruvic acid, oxalic acid, glycolic acid, salicylic acid, pyranotropic acids such as glucuronic acid or galacturonic acid, α-hydroxy acids such as citric acid or tartaric acid, amino acids such as aspartic acid or glutamic acid, aromatic acids such as benzoic acid or cinnamic acid, sulfonic acids such as p-toluenesulfonic acid or ethanesulfonic acid, etc.

[0146] Similarly, if a particular compound is an acid, then the desired pharmaceutically acceptable salt can be prepared by any suitable method, for example, by treating the free acid with inorganic or organic bases such as amines (primary, secondary, or tertiary amines), alkali metal hydroxides, or alkaline earth metal hydroxides. Illustrative examples of suitable salts include organic salts derived from amino acids such as L-glycine, L-lysine, and L-arginine, ammonia, primary, secondary, and tertiary amines, and cyclic amines such as hydroxyethylpyrrolidine, piperidine, morpholine, or piperazine; and inorganic salts derived from sodium, calcium, potassium, magnesium, manganese, iron, copper, zinc, aluminum, and lithium.

[0147] It should also be understood that the compounds disclosed herein may exist in unsolvable, solvated (e.g., hydrated) and solid (e.g., crystalline or polycrystalline) forms, and this disclosure is intended to cover all such forms.

[0148] As used herein, the term "solvent" or "solventized form" refers to a solvation form containing stoichiometric or non-stoichiometric amounts of a solvent. Some compounds have a tendency to trap a fixed molar ratio of solvent molecules in a crystalline solid state, thereby forming a solvate. If the solvent is water, the formed solvate is a hydrate; and if the solvent is an alcohol, the formed solvate is an alcohol. Hydrates are formed by combining one or more water molecules with a molecule of a substance, wherein the water molecule remains in the H₂O state. Examples of solvents that form solvates include, but are not limited to, water, isopropanol, ethanol, methanol, DMSO, ethyl acetate, acetic acid, and ethanolamine.

[0149] This disclosure is also intended to include all isotopes of atoms in the compounds. Isotopes of atoms include atoms having the same number of atoms but different mass numbers. For example, unless otherwise stated, hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine, bromine, or iodine in the compounds of this disclosure are intended to also include their isotopes, such as, but not limited to, those of other atoms. 1 H, 2 H, 3 H, 11 C 12 C 13 C 14 C 14 N、 15 N、 16 O、 17 O、 18 O、 31 P, 32 P, 32 S, 33 S, 34 S, 36 S, 17 F, 18 F, 19 F, 35 Cl、 37 Cl、 79 Br、 81 Br、 124 I, 127 I and 131 I. In some embodiments, hydrogen includes protium, deuterium, and tritium. In some embodiments, carbon includes... 12 C and 13 C.

[0150] The compounds provided herein, or their pharmaceutically acceptable salts, may contain one or more asymmetric centers, thus yielding enantiomers, diastereomers, and other stereoisomers, which can be defined by absolute stereochemistry as (R)- or (S)- forms or (D)- or (L)- forms of amino acids, or by relative configuration as rel-(R)- or rel-(S)- forms. This disclosure includes all such possible isomers as well as their racemic and optically pure forms. Optically active (+) and (-), (R)- and (S)- or (D)- and (L)- isomers can be prepared using chiral synthons or chiral reagents, or resolved by conventional techniques such as chromatography and fractional crystallization. Conventional techniques for preparing and separating individual enantiomers include chiral synthesis from suitable optically pure precursors or resolution of racemates (or racemates of salts or derivatives) using, for example, chiral high-performance liquid chromatography (HPLC). When a compound is represented in its chiral form, it should be understood that the embodiments described include, but are not limited to, specific diastereomers or enantiomer-enriched forms. Where chirality is present but not specified, it should be understood that the embodiments are intended to include specific diastereomers or enantiomer-enriched forms; or racemic or scalemic mixtures of such compounds.

[0151] The term "stereoisomer" refers to a compound containing identical atoms bonded by the same bonds but having different three-dimensional structures that are not interchangeable. This disclosure considers various stereoisomers and mixtures thereof and includes "enantiomers," which are two stereoisomers whose molecules are non-overlapping mirror images of each other.

[0152] The term "enantiomer" refers to a pair of stereoisomers that are non-overlapping mirror images of each other. A 1:1 mixture of a pair of enantiomers is a "racemic" mixture. A mixture of enantiomers in a non-1:1 ratio is a "non-racemic" mixture.

[0153] The term "diastereomer" refers to a stereoisomer that has at least two asymmetric atoms but is not a mirror image of each other.

[0154] The terms "tautomer" or "tautomer form" refer to structural isomers of different energies that can interconvert via low energy barriers. The presence and concentration of isomer forms will depend on the environment in which the compound is situated and can vary depending on, for example, whether the compound is a solid or whether it is in an organic or aqueous solution. For example, proton tautomers (also known as proton-loving tautomers) include interconversions via proton migration, such as keto-enol, amide-imino, lactam-lactamimide, imine-enamine isomerization, and cyclic forms, in which protons can occupy two or more positions in a heterocyclic system. Valence tautomers include interconversions that occur through the rearrangement of some bonding electrons. Tautomers can be in equilibrium or spatially locked into one form through appropriate substitution. Unless otherwise stated, the compounds of this disclosure identified by name or structure as a particular tautomer form are intended to include other tautomer forms.

[0155] When the compounds provided herein contain olefinic double bonds or other geometrically asymmetric centers, these compounds include E and Z geometric isomers unless otherwise stated.

[0156] Synthesis method

[0157] The compounds presented in this article can be prepared using any known organic synthesis technique and can be synthesized according to any of the many possible synthetic routes.

[0158] The reactions used to prepare the compounds of this disclosure can be carried out in suitable solvents, which can be readily selected by those skilled in the art of organic synthesis. Suitable solvents are substantially non-reactive with the starting materials (reactants), intermediates, or products at the temperature at which the reaction takes place, said temperature being, for example, from the freezing point of the solvent to its boiling point. A given reaction can be carried out in one solvent or a mixture of more than one solvent. Depending on the specific reaction step, a suitable solvent for that particular reaction step can be selected by those skilled in the art.

[0159] The preparation of the compounds disclosed herein may involve the protection and deprotection of various chemical groups. Those skilled in the art can readily determine the need for protection and deprotection, as well as the selection of appropriate protecting groups. The chemistry of protecting groups can be found, for example, in T.W. Greene and P.G. M. Wuts, *Protective Groups in Organic Synthesis*, 3rd ed., Wiley & Sons, Inc., New York (1999); P. Kocienski, *Protecting Groups*, Georg Thieme Verlag, 2003; and Peter G.M. Wuts, *Greene's Protective Groups in Organic Synthesis*, 5th ed., Wiley, 2014, all of which are incorporated herein by reference in their entirety.

[0160] The reaction can be monitored using any suitable method known in the art. For example, product formation can be monitored by spectroscopic methods, such as nuclear magnetic resonance spectroscopy (e.g., 1 H or 13 C) Infrared spectroscopy, spectrophotometry (e.g., UV-Vis), mass spectrometry, or chromatographic methods such as high-performance liquid chromatography (HPLC), liquid chromatography-mass spectrometry (LCMS), or thin-layer chromatography (TLC). Those skilled in the art can purify compounds using a variety of methods, including high-performance liquid chromatography (HPLC) (“Preparative LC-MS Purification: Improved Compound Specific Method Optimization” Karl F. Blom, Brian Glass, Richard Sparks, Andrew P. Combs, *J. Combi. Chem.*, 2004, 6(6), 874-883, which is incorporated herein by reference in its entirety) and normal-phase silica gel chromatography.

[0161] Pharmaceutical Composition

[0162] In another aspect, pharmaceutical compositions comprising one or more compounds of the present disclosure or pharmaceutically acceptable salts thereof are provided. In some embodiments, the pharmaceutical compositions of the present disclosure comprise a first compound of formula (I) or a pharmaceutically acceptable salt thereof and one or more other compounds of the same formula, wherein the first compound and the other compounds are not the same molecule.

[0163] In another aspect, a pharmaceutical composition comprising one or more compounds disclosed herein or pharmaceutically acceptable salts thereof and at least one pharmaceutically acceptable excipient is provided.

[0164] In some embodiments, the pharmaceutical compositions of this disclosure comprise a therapeutically effective amount of one or more of the compounds of this disclosure or a pharmaceutically acceptable salt thereof.

[0165] In some embodiments, the pharmaceutical compositions of this disclosure comprise a therapeutically effective amount of one or more of the compounds of this disclosure or pharmaceutically acceptable salts thereof and at least one pharmaceutically acceptable excipient.

[0166] As used herein, the term "therapeutic effective amount" refers to the amount of a molecule, compound, or composition comprising said molecule or compound that treats, improves, or prevents a diagnosed disease or symptom or exhibits a detectable therapeutic or inhibitory effect. The effect can be detected by any assay method known in the art. The precise effective amount for a subject will depend on the subject's weight, body type, and health status; the nature and severity of the symptom; the rate of administration; the choice of the therapeutic agent or combination of therapeutic agents used for administration; and the prescribing physician's judgment. The therapeutic effective amount for a given situation can be determined through routine laboratory testing within the skill and judgment of a clinician.

[0167] As used herein, the term "pharmaceutical composition" refers to a formulation containing a molecule or compound of the present disclosure in a form suitable for administration to a subject. Pharmaceutical compositions include those suitable for oral, rectal, topical, parenteral (including subcutaneous, intramuscular, and intravenous) administration, sublingual, ocular, transdermal, or intranasal administration; however, in any given case, the most suitable route will depend on the specific host and the nature and severity of the symptom to which the active ingredient is administered. Pharmaceutical compositions can be conveniently present in unit dosage forms and prepared by any method well known in the pharmaceutical field.

[0168] As used herein, the term "pharmaceuticalally acceptable excipient" means an excipient that can be used to prepare pharmaceutical compositions that are generally safe and non-toxic and are not biologically or otherwise undesirable, and includes excipients acceptable for veterinary and human use. As used herein, "pharmaceuticalally acceptable excipient" includes one or more such excipients. The term "pharmaceuticalally acceptable excipient" also covers "pharmaceuticalally acceptable carriers" and "pharmaceuticalally acceptable diluents."

[0169] The specific excipients used will depend on the manner and purpose of applying the compounds disclosed herein. Solvents are generally selected based on those considered safe by those skilled in the art for application to mammals, including humans. Generally, safe solvents are non-toxic aqueous solvents, such as water and other non-toxic solvents soluble or miscible in water. Suitable aqueous solvents include water, ethanol, propylene glycol, polyethylene glycol (e.g., PEG 400, PEG 300), and mixtures thereof.

[0170] In some embodiments, suitable excipients may include buffers such as phosphates, citrates, and other organic acids; antioxidants, including ascorbic acid and methionine; preservatives (e.g., octadecyl dimethyl benzyl ammonium chloride; hexamethyl diammonium chloride; benzalkonium chloride, benzyl chloride; phenol, butanol, or benzyl alcohol; alkyl esters of p-hydroxybenzoate, such as methylparaben or propylparaben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) polyphenols. Peptides; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers, such as polyvinylpyrrolidone; amino acids, such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates, including glucose, mannose, or dextrin; chelating agents, such as EDTA; sugars, such as sucrose, mannitol, trehalose, or sorbitol; salt-forming counterions, such as sodium; metal complexes (e.g., zinc-protein complexes); and / or nonionic surfactants, such as TWEEN. TM PLURONICS TM Or polyethylene glycol (PEG).

[0171] In some embodiments, suitable excipients may include one or more stabilizers, surfactants, wetting agents, lubricants, emulsifiers, suspending agents, preservatives, antioxidants, opacifiers, slip agents, processing aids, colorants, sweeteners, flavoring agents, and other known additives to provide optimal presentation of the pharmaceutical product (i.e., the compound of this disclosure or a pharmaceutical composition thereof) or to aid in the manufacture of a pharmaceutical product (i.e., a pharmaceutical preparation). The active pharmaceutical ingredient may also be encapsulated in microcapsules, for example, prepared by coagulation techniques or by interfacial polymerization, such as hydroxymethyl cellulose or gelatin microcapsules and poly(methyl methacrylate) microcapsules, respectively, in colloidal drug delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles, and nanocapsules) or in crude emulsions. This technique is disclosed in Remington's Pharmaceutical Sciences, 16th edition, Osol, A. (1980). Liposomes are small vesicles composed of various types of lipids, phospholipids, and / or surfactants, which can be used to deliver drugs (e.g., the compounds disclosed herein and optional chemotherapeutic agents) to mammals, including humans. The components of liposomes are typically arranged in a bilayer form, similar to the lipid arrangement of biological membranes.

[0172] The pharmaceutical compositions provided herein may be in any form that allows the composition to be administered to subjects, including but not limited to humans, and formulated to be compatible with the intended route of administration.

[0173] Multiple routes of administration are considered for the pharmaceutical compositions provided herein, and therefore, the pharmaceutical compositions provided herein may be supplied in bulk or unit dosage forms depending on the intended route of administration. For example, for oral, buccal, and sublingual administration, powders, suspensions, granules, tablets, pills, capsules, capsule tablets, and capsule sachets are acceptable as solid dosage forms, and emulsions, syrups, elixirs, suspensions, and solutions are acceptable as liquid dosage forms. For injectable administration, emulsions and suspensions are acceptable as liquid dosage forms, and powders suitable for reconstitution with appropriate solutions are acceptable as solid dosage forms. For inhalation administration, solutions, sprays, dry powders, and aerosols are acceptable dosage forms. For topical (including buccal and sublingual) or transdermal administration, powders, sprays, ointments, pastes, creams, lotions, gels, solutions, and patches are acceptable dosage forms. For vaginal administration, vaginal suppositories, tampons, creams, gels, pastes, foams, and sprays are acceptable dosage forms.

[0174] In some embodiments, the pharmaceutical compositions disclosed herein may be in the form of an oral administration formulation.

[0175] In some embodiments, the pharmaceutical compositions of this disclosure may be in the form of tablet formulations. For example, suitable pharmaceutically acceptable excipients for tablet formulations include inert diluents such as lactose, sodium carbonate, calcium phosphate, or calcium carbonate; granulating and disintegrants such as corn starch or alginate; binders such as starch; lubricants such as magnesium stearate, stearic acid, or talc; preservatives such as ethylparaben or propylparaben; and antioxidants such as ascorbic acid. Tablet formulations may be uncoated or coated to modulate their disintegration and subsequent absorption of the active ingredient in the gastrointestinal tract, or to improve their stability and / or appearance; in either case, conventional coating agents and procedures well known in the art are used.

[0176] In some embodiments, the pharmaceutical compositions of this disclosure may be in the form of hard gelatin capsules, wherein the active ingredient is mixed with an inert solid diluent, such as calcium carbonate, calcium phosphate or kaolin; or in the form of soft gelatin capsules, wherein the active ingredient is mixed with water or oil, such as peanut oil, liquid paraffin or olive oil.

[0177] In some embodiments, the pharmaceutical compositions of this disclosure may be in the form of an aqueous suspension, which typically contains an active ingredient in fine powder form and one or more suspending agents, such as sodium carboxymethyl cellulose, methyl cellulose, hydroxypropyl methyl cellulose, sodium alginate, polyvinylpyrrolidone, tragacanth gum, and gum arabic; dispersants or wetting agents, such as lecithin or condensation products of ethylene oxide and fatty acids (e.g., polyoxyethylene stearate), or condensation products of ethylene oxide and long-chain fatty alcohols, such as heptadecaethyleneoxy cetyl alcohol, or condensation products of ethylene oxide and esters derived from fatty acids and hexitols, such as polyoxyethylene sorbitan monooleate, or condensation products of ethylene oxide and esters derived from fatty acids and hexitol anhydrides, such as polyvinyl sorbitan monooleate. Aqueous suspensions may also contain one or more preservatives (e.g., ethylparaben or propylparaben), antioxidants (e.g., ascorbic acid), colorants, flavorings, and / or sweeteners (e.g., sucrose, saccharin, or aspartame).

[0178] In some embodiments, the pharmaceutical compositions of this disclosure may be in the form of an oily suspension, which typically contains a suspended active ingredient in a vegetable oil (e.g., peanut oil, olive oil, sesame oil, or coconut oil) or a mineral oil (e.g., liquid paraffin). The oily suspension may also contain a thickener, such as beeswax, hard paraffin, or cetyl alcohol. Sweeteners and flavoring agents as described above may be added to provide a palatable oral formulation. These compositions may be preserved by adding antioxidants such as ascorbic acid.

[0179] In some embodiments, the pharmaceutical compositions of this disclosure may be in the form of an oil-in-water emulsion. The oil phase may be a vegetable oil, such as olive oil or peanut oil; or a mineral oil, such as liquid paraffin; or any mixture of these oils. Suitable emulsifiers may be, for example, naturally occurring gums, such as gum arabic or tragacanth; naturally occurring phospholipids, such as soybean, lecithin, esters or metaesters derived from fatty acids and hexitan anhydrides (e.g., sorbitan monooleate), and condensation products of said metaesters with ethylene oxide, such as polyoxyethylene sorbitan monooleate. The emulsion may also contain sweeteners, flavoring agents, and preservatives.

[0180] In some embodiments, the pharmaceutical compositions provided herein may be in the form of syrups and elixirs, and may contain sweeteners such as glycerin, propylene glycol, sorbitol, aspartame, or sucrose; modifiers; preservatives; flavoring agents and / or coloring agents.

[0181] In some embodiments, the pharmaceutical compositions of this disclosure may be in the form of an injectable formulation.

[0182] In some embodiments, the pharmaceutical compositions of this disclosure may be in the form of sterile injectable formulations, such as sterile injectable aqueous or oily suspensions. This suspension may be formulated using suitable dispersants or wetting agents and suspending agents mentioned above, according to known techniques. The sterile injectable formulation may also be a sterile injectable solution or suspension in a non-toxic, parenteral-acceptable diluent or solvent, such as a solution in 1,3-butanediol, or prepared as a lyophilized powder. Acceptable mediators and solvents that can be used are water, Ringer's solution, and isotonic sodium chloride solution. Additionally, sterile non-volatile oils may be routinely used as solvents or suspension media. For this purpose, any mild non-volatile oil may be used, including synthetic monoglycerides or diglycerides. Furthermore, fatty acids such as oleic acid may also be used in the preparation of injectables.

[0183] In some embodiments, the pharmaceutical compositions disclosed herein may be in the form of an inhalation preparation.

[0184] In some embodiments, the pharmaceutical compositions of this disclosure may be in the form of aqueous and non-aqueous (e.g., in fluorocarbon propellants) aerosols containing any suitable solvent and optional other compounds, such as, but not limited to, stabilizers, antimicrobial agents, antioxidants, pH adjusters, surfactants, bioavailability modifiers, and combinations thereof. Carriers and stabilizers vary depending on the requirements of the specific compound, but generally include nonionic surfactants (Tween, Pluronics, or polyethylene glycol), harmless proteins such as serum albumin, sorbitol esters, oleic acid, lecithin, amino acids such as glycine, buffers, salts, sugars, or sugar alcohols.

[0185] In some embodiments, the pharmaceutical compositions of this disclosure may be in the form of a topical or transdermal formulation. In some embodiments, the pharmaceutical compositions provided herein may be in the form of creams, ointments, gels, and aqueous or oily solutions or suspensions, which may generally be obtained by formulation of the active ingredient and conventional, topically acceptable excipients, such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth gum, cellulose derivatives, polyethylene glycol, silicone, bentonite, silicic acid, talc, and zinc oxide, or mixtures thereof.

[0186] In some embodiments, the pharmaceutical compositions provided herein can be formulated for ophthalmic application. In some embodiments, the pharmaceutical compositions provided herein can be in the form of ophthalmic formulations, such as ophthalmic ointments, powders, solutions, etc. In some embodiments, the ophthalmic formulations are prepared using a suitable buffer system at an appropriate pH.

[0187] In addition to the representative dosage forms mentioned above, pharmaceutically acceptable excipients and carriers are generally known to those skilled in the art and are therefore included in this disclosure. Such excipients and carriers are described, for example, in Remington's Pharmaceutical Sciences, Mack Pub. Co., New Jersey (1991); Remington: The Science and Practice of Pharmacy, University of the Science of Philadelphia, ed., 21st ed., LWW (2005), which are incorporated herein by reference.

[0188] The dosage regimens for the compounds described herein will vary depending on known factors such as the pharmacodynamic properties of the specific agent and its route of administration; the recipient's species, age, sex, health status, medical condition, and weight; the nature and severity of symptoms; the types of concurrent treatments; the frequency of treatment; the route of administration; the patient's renal and hepatic function; and the desired effect. A physician or veterinarian can determine and prescribe an effective amount of medication needed to prevent, combat, or halt the progression of the disease.

[0189] In some embodiments, the pharmaceutical compositions of this disclosure can be formulated such that the dosage can be administered in the range of 0.001-1000 mg / kg body weight / day, for example, dosages of 0.01-800 mg / kg body weight / day, 0.01-700 mg / kg body weight / day, 0.01-600 mg / kg body weight / day, 0.01-500 mg / kg body weight / day, 0.01-400 mg / kg body weight / day, 0.01-300 mg / kg body weight / day, 0.1-200 mg / kg body weight / day, 0.1-150 mg / kg body weight / day. The dosages of the compounds described herein, or their pharmaceutically acceptable salts, are as follows: 0.1-100 mg / kg body weight / day, 0.5-100 mg / kg body weight / day, 0.5-80 mg / kg body weight / day, 0.5-60 mg / kg body weight / day, 0.5-50 mg / kg body weight / day, 1-50 mg / kg body weight / day, 1-45 mg / kg body weight / day, 1-40 mg / kg body weight / day, 1-35 mg / kg body weight / day, 1-30 mg / kg body weight / day, 1-25 mg / kg body weight / day. In some instances, dose levels below the lower limit of the foregoing ranges may be sufficient, while in other cases, larger doses may be used without causing any harmful side effects, provided that such larger doses are first divided into several smaller doses for administration throughout the day. For further information on the route of administration and dosage regimen, see Chapter 25.3 of Volume 5 of Comprehensive Medicinal Chemistry (Corwin Hansch; Chairman of the Editorial Board), Pergamon Press, 1990, which is hereby incorporated herein by reference.

[0190] In some embodiments, the pharmaceutical compositions of this disclosure may be formulated into a single dosage form. The amount of the compounds provided herein in a single dosage form will vary depending on the subject being treated and the specific method of administration.

[0191] In some embodiments, suitable dosage forms may contain about 1 mg to about 1000 mg of active ingredient per dose unit. In these pharmaceutical compositions, the active ingredient will typically be present in an amount of about 0.1-95% by weight based on the total weight of the composition.

[0192] In some embodiments, the pharmaceutical compositions of this disclosure can be formulated as short-acting, rapid-release, long-acting, and sustained-release formulations. Therefore, the pharmaceutical formulations of this disclosure can also be formulated for controlled-release or slow-release applications.

[0193] In some embodiments, a dose of the compound or pharmaceutical composition provided herein is administered to the subject daily, every other day, every few days, every three days, once a week, twice a week, three times a week, or every two weeks. If desired, the effective daily dose of the active compound may be administered individually at appropriate intervals throughout the day in two, three, four, five, six, or more sub-dose doses, optionally in unit dosage form. In some embodiments, the compound or pharmaceutical composition provided herein is administered at doses for 2, 3, 5, 7, 14, 21 days, 1 month, 2 months, 2.5 months, 3 months, 4 months, 5 months, 6 months, or longer.

[0194] In another aspect, a veterinary composition is provided comprising one or more molecules or compounds disclosed herein, or pharmaceutically acceptable salts thereof, and a veterinary carrier. The veterinary carrier is a substance suitable for the purpose of administering the composition and may be an inert or veterinarily acceptable solid, liquid, or gaseous substance compatible with the active ingredient. These veterinary compositions may be administered parenterally, orally, or via any other desired route.

[0195] Pharmaceutical or veterinary compositions may be packaged in various ways depending on the method of administration. For example, articles for dispensing may include containers in which a suitable form of composition is placed. Suitable containers are well known to those skilled in the art and include materials such as bottles (plastic and glass), capsules, ampoules, plastic bags, metal tubes, etc. Containers may also include tamper-evident components to prevent easy access to the contents of the package. Additionally, the container is affixed with a label describing the contents. The label may also include appropriate warnings. Compositions may be packaged in single-dose or multi-dose containers, such as sealed ampoules and vials, and may be stored under lyophilized (freeze-dried) conditions, requiring only immediate addition of a sterile liquid carrier, such as water, for injection before use. Ready-to-use injectable solutions and suspensions are prepared from sterile powders, granules, and tablets of the types described above.

[0196] In some embodiments, the pharmaceutical compositions of this disclosure comprising one or more compounds provided herein or pharmaceutically acceptable salts thereof further comprise one or more additional therapeutically active agents.

[0197] The additional therapeutic agents have complementary activities to the compounds presented herein, ensuring that they do not adversely affect each other. These agents are appropriately combined in amounts effective for the intended purpose.

[0198] In some embodiments, the additional therapeutic agent is selected from beta blockers, alpha agonists, carbonic anhydrase inhibitors, prostaglandin-like compounds, miotics, cholinergic agents, or adrenaline compounds.

[0199] Beta-blockers reduce aqueous humor production. Examples include levobenolol. Timolol Betalol Hemetrol

[0200] Alpha-agonists reduce aqueous humor production and increase drainage. Examples include adapalene. and brimonidine

[0201] Carbonic anhydrase inhibitors reduce aqueous humor production. Examples include dzoxol. and brinzoamine

[0202] Prostaglandins and prostaglandin-like compounds increase aqueous humor outflow. Examples include latanoprost. Bimatoprost Travitan TM ).

[0203] Miotics or cholinergics increase the outflow of aqueous humor. Examples include pilocarpine. and carbacholine

[0204] Adrenaline compounds, such as dipiformin It also increases the outflow of aqueous humor.

[0205] One or more additional therapeutic agents may be administered simultaneously or sequentially with the compounds provided herein. Sequential administration includes administration before or after the compounds provided herein. In some embodiments, one or more additional therapeutic agents may be administered in the same composition as the compounds provided herein. In other embodiments, a time interval may exist between the administration of the additional therapeutic agent and the compounds provided herein.

[0206] In some embodiments, administering additional therapeutic agents together with the compounds provided herein can achieve lower doses of other therapeutic agents and / or administration at lower frequency intervals.

[0207] Treatment

[0208] The compounds disclosed herein and pharmaceutical compositions comprising them are capable of inhibiting ROCK and are therefore suitable for use in subjects in need of inhibiting ROCK activity and for the prevention or treatment of ROCK-related conditions.

[0209] In another aspect, this disclosure provides a method for treating ROCK-related conditions, comprising administering to a subject in need an effective amount of the compound provided herein or a pharmaceutically acceptable salt or pharmaceutical composition thereof.

[0210] As used herein, the terms “treating” or “therapy” are intended to have the normal meaning of managing a disease in order to completely or partially alleviate one, some, or all of its symptoms or to correct or compensate for the underlying pathology, thereby achieving a beneficial or desired clinical outcome. For the purposes of this disclosure, beneficial or desired clinical outcomes include, but are not limited to, relief of symptoms, reduction of disease severity, stabilization of the disease state (i.e., no worsening), delay or slowing of disease progression, improvement or mitigation of the disease condition, and remission (whether partial or complete), whether detectable or undetectable. “Treatment” may also mean an extension of survival compared to expected survival without treatment. People in need of therapy include those who already have symptoms or conditions, those who are susceptible to symptoms or conditions, or those who should prevent symptoms or conditions.

[0211] As used herein, the term “preventing / prevention / prophylaxis” is intended to have its normal meaning and includes primary and secondary prevention of disease progression, in which case the disease has already progressed and the patient is temporarily or permanently protected from the aggravation or worsening of the disease or the development of new symptoms associated with the disease.

[0212] In some embodiments, the compounds provided herein or their pharmaceutically acceptable salts and compositions may be used to treat a variety of rock-related conditions, including cancer, cardiovascular disease, autoimmune disease, eye disease, metabolic syndrome, acute respiratory distress syndrome, kidney disease, overactive bladder, epilepsy, migraine, diabetes, high-altitude pulmonary edema, and mental disorders.

[0213] In some embodiments, the compounds provided herein, or their pharmaceutically acceptable salts and compositions, can be used to treat eye diseases, including glaucoma, ocular hypertension, retinal diseases, including wet AMD, dry AMD, and DME; bone diseases, including osteoporosis and osteoarthritis; vascular diseases, including cerebral vasospasm, coronary vasospasm, hypertension, pulmonary hypertension, high-altitude essential hypertension, sudden death syndrome, angina pectoris, myocardial infarction, restenosis, stroke, hypertensive vascular disease, heart failure, cardiac graft vascular disease, atherosclerosis, arterial occlusion, peripheral artery disease, peripheral circulatory disorders, and venous graft disease; lung diseases, including chronic obstructive pulmonary disease (COPD) and asthma; and nerve diseases. Diseases including spinal cord injury, dementia, Alzheimer's disease, Parkinson's disease, neuronal degeneration, multiple sclerosis, depression, attention deficit hyperactivity disorder and neuropathic pain, multiple sclerosis, amyotrophic lateral sclerosis; neovascularization; cancers including astrocytoma, breast cancer, cervical cancer, colorectal cancer, endometrial cancer, esophageal cancer, stomach cancer, head and neck cancer, hepatocellular carcinoma, laryngeal cancer, lung cancer, oral cancer, ovarian cancer, prostate cancer and thyroid cancer and sarcoma; obesity; urinary tract diseases including overactive bladder (OAB) and benign prostatic hyperplasia (BPH); erectile dysfunction; acute and chronic pain; asthma; spinal cord injury; rheumatoid arthritis; psoriasis; viral infections; cardioprotection.

[0214] In some embodiments, the eye disease that can be treated with the compounds provided herein or their pharmaceutically acceptable salts and compositions is glaucoma.

[0215] The concentration and route of administration to the subject will vary depending on the ROCK-related condition to be treated. In some embodiments, the administration is carried out via a route selected from the group consisting of: parenteral administration, intraperitoneal administration, intradermal administration, intracardiac administration, intraventricular administration, intracranial administration, intraspinal administration, intrasynovial administration, intrathecal administration, intrathecal administration, intramuscular injection, intravitreal injection, intravenous injection, intra-arterial injection, oral administration, buccal administration, sublingual administration, transdermal administration, local administration, intratracheal administration, rectal administration, subcutaneous administration, and ocular administration.

[0216] Example

[0217] The general methods of this disclosure are further illustrated below. The compounds of this disclosure can be prepared by methods known in the art. Detailed methods for preparing preferred compounds of this disclosure are described below. However, these are in no way limiting the methods for preparing the compounds of this disclosure.

[0218] Synthesis Example

[0219] For illustrative purposes, the following examples are included. The examples provided herein describe the synthesis of the compounds disclosed herein and the intermediates used to prepare said compounds. However, it should be understood that these examples are not limiting of this disclosure and are intended only to illustrate the methods of practicing this disclosure. Those skilled in the art will recognize that the described chemical reactions can be readily applied to the preparation of a variety of other compounds of this disclosure, and alternative methods for preparing the compounds of this disclosure are considered to be within the scope of this disclosure. For example, non-exemplary compounds according to this disclosure can be successfully synthesized by modifications obvious to those skilled in the art, such as by appropriately protecting interfering groups, by utilizing other suitable reagents and building blocks known in the art besides said reagents and building blocks, and / or by conventional modifications to the reaction conditions. Furthermore, those skilled in the art will understand that individual steps or different batches of compounds described herein can be combined. Alternatively, other reactions disclosed herein or known in the art will be considered suitable for the preparation of other compounds of this disclosure. Therefore, the following description is not intended to limit the scope of this disclosure, but is defined by the appended claims.

[0220] The abbreviations of the chemical reagents used in the synthesis of the compounds provided in this article are listed below:

[0221]

[0222]

[0223] Synthesis of intermediates

[0224] Synthesis of intermediate I-1

[0225]

[0226] Step 1: At room temperature and under a nitrogen atmosphere, a solution of p-cyanomethylbenzoic acid I-1a (20 g, 124.10 mmol, 1 equivalent) and CDI (22.14 g, 136.51 mmol, 1.1 equivalent) in THF (50 mL) was stirred for 3 hours. At 0 °C and under a nitrogen atmosphere, H₂O (45 mL) containing NaBH₄ (14.08 g, 372.30 mmol, 3 equivalent) was added dropwise to the mixture. The resulting mixture was stirred overnight at room temperature and under a nitrogen atmosphere. The reaction was quenched with water at room temperature. The mixture was extracted with ethyl acetate (3 x 500 mL). The combined organic layers were dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using CH2Cl2 / EtOAc (8:1, v / v) elution to obtain 2-[4-(hydroxymethyl)phenyl]acetonitrile I-1b (10.9 g, 59.67%, crude product), which was a pale yellow liquid. 1H NMR (CD3OD, 400MHz) δ3.87–3.91(2H,m),4.62(2H,s),7.37(4H,q).

[0227] Step 2: At 0°C and under a nitrogen atmosphere, TBSCl (13.39 g, 88.87 mmol, 1.2 equivalent) was added in portions to a stirred mixture of alcohol I-1b (10.9 g, 74.06 mmol, 1 equivalent) and triethylamine (22.48 g, 222.18 mmol, 3 equivalent), and DMAP (0.09 g, 0.741 mmol, 0.01 equivalent) in 100 mL of DCM. The resulting mixture was stirred for 4 hours at room temperature under a nitrogen atmosphere. The mixture was diluted with water (260 mL). The mixture was extracted with CH2Cl2 (3 x 100 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC (PE / EA = 6:1, v / v) to obtain 2-(4-(((tert-butyldimethylsilyl)oxy)methyl)phenyl)acetonitrile I-1c (14.0 g, 72.31%), which was a pale yellow liquid. 1 H NMR (CD3OD, 400MHz) δ0.13(6H,s),0.96(9H,s),3.89(2H,s),4.77(2H,s),7.31–7.40(4H,m).

[0228] Step 3: Sodium hydride (1.93 g, 80.33 mmol, 1 equivalent) was added in portions to a stirred solution of nitrile I-1c (14.0 g, 53.55 mmol, 1 equivalent) in THF (20 mL). The resulting mixture was stirred for 1 hour at 0 °C under a nitrogen atmosphere. Dimethyl carbonate (19.29 g, 214.20 mmol, 4 equivalents) was added in portions to the above mixture at 0 °C under a nitrogen atmosphere. The resulting mixture was stirred overnight at room temperature under a nitrogen atmosphere. The reaction was quenched with water (200 mL) at room temperature. The mixture was extracted with ethyl acetate (3 x 250 mL). The combined organic layers were dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE / EA = 6:1, v / v) to obtain methyl 2-(4-{[(tert-butyldimethylsilyl)oxy]methyl}phenyl)-2-cyanoacetate I-1d (12.6 g, 73.65%), which was a pale yellow liquid. 1 H NMR (CD3OD, 400MHz) δ0.13(6H,s),0.97(9H,s),3.78(3H,s),4.78(2H,s),7.37–7.47(4H,m).

[0229] Step 4: At room temperature, add 150 mL of methanol containing ester I-1d (12.6 g, 39.44 mmol, 1 equivalent), Boc₂O (34.44 g, 157.76 mmol, 4 equivalents), and CoCl₂·6H₂O (24.42 g, 102.55 mmol, 2.6 equivalents) to a 250 mL round-bottom flask. At 0 °C, add NaBH₄ (11.94 g, 315.53 mmol, 8 equivalents) to the mixture. Stir the resulting mixture for 15 hours at room temperature under a nitrogen atmosphere. Dry the combined organic layers with anhydrous Na₂SO₄. After filtration, concentrate the filtrate under reduced pressure. The residue was purified by silica gel column chromatography using PE / EA (0-5.5%) elution to obtain methyl 3-[(tert-butoxycarbonyl)amino]-2-(4-{[(tert-butyldimethylsilyl)oxy]methyl}phenyl)propionate I-1e (7.1 g, 42.51%), which was a pale yellow oil. 1 H NMR(DMSO-d6,400MHz)δ0.08(6H,s),0.91(9H,s),1.34(9H,s),3.25(1H,dt),3.48(1H ,ddd),3.59(3H,s),3.84(1H,t),4.68(2H,s),6.93(1H,t),7.22(2H,d),7.27(2H,d).

[0230] Step 5: At 60°C under a nitrogen atmosphere, a solution of methanol (10 mL) containing ester I-1e (10.1 g, 23.84 mmol, 1 equivalent) and H2O (6 mL) containing LiOH·H2O (2.00 g, 47.68 mmol, 2 equivalents) was stirred overnight. The resulting mixture was concentrated under reduced pressure. The mixture was acidified to pH 6 with HCl (aqueous solution). The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, methanol / water, 10% to 50% gradient, 40 min; detector, UV 220 nm. This yielded 3-[(tert-butoxycarbonyl)amino]-2-[4-(hydroxymethyl)phenyl]propionic acid I-1 (3.58 g, 50.84%) as a grayish-white solid. LCMS: m / z (ESI), [M+Na] + =318.00.

[0231] Synthesis of intermediate I-2

[0232]

[0233] Step 1: At 0 °C, sodium hydride (0.54 g, 22.42 mmol, 1.1 equivalent) was added to a solution of 2-(3-methoxyphenyl)acetonitrile I-2a (3 g, 20.38 mmol, 1 equivalent) in THF (50 mL). The mixture was stirred for 50 min. Dimethyl carbonate (14.69 g, 163.07 mmol, 8 equivalent) was added, and the mixture and the resulting mixture were heated to RT and stirred for 2 h. The reaction mixture was quenched with water (50 mL) and extracted with DCM (3 x 50 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using PE / EA (12:1, v / v) elution to give methyl 2-cyano-2-(3-methoxyphenyl)acetate I-2b (2.84 g, 67.89%) as a yellow oil. 1 HNMR (400MHz, DMSO-d6) δ3.76(d,6H),5.63(s,1H),6.81–7.11(m,3H),7.39(t,1H).

[0234] Step 2: At room temperature and under a nitrogen atmosphere, a mixture of ester I-2b (800 mg, 3.90 mmol, 1 equivalent), Boc2O (1701.62 mg, 7.80 mmol, 2 equivalents), CoCl2·6H2O (1113.06 mg, 4.68 mmol, 1.2 equivalents), and NaBH4 (368.72 mg, 9.75 mmol, 2.5 equivalents) in methanol (20 mL) was stirred overnight. The resulting mixture was diluted with water (30 mL), extracted with CH2Cl2 (3 x 40 mL), and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC (PE / EA = 2:1, v / v) to give methyl 3-[(tert-butoxycarbonyl)amino]-2-(3-methoxyphenyl)propionate I-2c (504 mg, 41.79%) as a pale yellow oil. LCMS:m / z(ESI),[M+Na] + =332.00.

[0235] Step 3: At room temperature, add 3 mL of THF and 3 mL of water containing ester I-2c (760 mg, 2.46 mmol, 1 equivalent) and LiOH (117.67 mg, 4.91 mmol, 2.0 equivalent) to a 50 mL round-bottom flask. Stir the resulting mixture for 3 hours at room temperature. Concentrate the mixture under reduced pressure. Acidify the mixture to pH 5 with HCl (1 M). Purify the residue by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, water / methanol, 0% to 100% gradient, 50 min; detector, UV 254 nm. Concentrate the resulting mixture under vacuum to give 200 mg (27.57%) of 3-[(tert-butoxycarbonyl)amino]-2-(3-methoxyphenyl)propionic acid as a brownish-yellow solid. LCMS: m / z (ESI), [M- t Bu] + =240.25.

[0236] Synthesis of intermediate I-3

[0237]

[0238] Step 1: At 0°C, sodium hydride (4.08 g, 102.02 mmol, 2 equivalents, 60%) was added to a solution of 2-(3-bromophenyl)acetonitrile I-3a (10 g, 51.01 mmol, 1 equivalent) in THF (200 mL). The mixture was stirred for 40 minutes. Dimethyl carbonate (18.38 g, 204.03 mmol, 4 equivalents) was added, and the mixture was heated to RT and stirred for 2 hours. The resulting mixture was quenched with water (500 mL) at 0°C. The resulting mixture was extracted with ethyl acetate (3 x 200 mL). The combined organic layers were washed with brine (2 x 200 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The crude product was recrystallized from DCM / PE (1:10 v / v, 500 mL) to give methyl 2-(3-bromophenyl)-2-cyanoacetate I-3b (6.5 g, 50.15%) as a white solid. LCMS: m / z (ESI), [MH] - =253.70. 1 H NMR(DMSO-d6,400MHz)δ3.43(3H,s),6.67(1H,ddd),6.94(1H,t),7.39(1H,d),7.86(1H,s).

[0239] Step 2: At 0°C and under air atmosphere, NaBH4 (4.91 g, 129.88 mmol, 6 equivalents) was added dropwise to a stirred mixture of ester I-3b (5.5 g, 21.65 mmol, 1 equivalent), CoCl2·6H2O (15.45 g, 64.94 mmol, 3 equivalents) and Boc2O (14.17 g, 64.94 mmol, 3 equivalents) in methanol (100 mL). The mixture was heated to RT and stirred for 2 hours. The resulting mixture was quenched with water (200 mL) at 0°C. The resulting mixture was filtered. The filter cake was washed with DCM (4 x 100 mL). The filtrate was extracted with CH2Cl2 (2 x 300 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using PE / EA (10:1, v / v) elution to give methyl 2-(3-bromophenyl)-3-[(tert-butoxycarbonyl)amino]propionate I-3c (4 g, 30.95%) as a yellow oil. LCMS: m / z (ESI), [M+H- t Bu] + =303.85.

[0240] Step 3: Under a nitrogen atmosphere, at 80 °C, a mixture of Pd(PPh3)4 (0.16 g, 0.14 mmol, 0.05 equivalent), Zn(CN)2 (0.20 g, 1.68 mmol, 0.6 equivalent), and ester I-3c (1 g, 2.79 mmol, 1 equivalent) in DMF (15 mL) was stirred for 4 hours. The resulting mixture was diluted with water (50 mL). The mixture was extracted with ethyl acetate (3 x 50 mL). The combined organic layers were washed with brine (3 x 70 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC (PE / EA = 3:1, v / v) to give methyl 3-[(tert-butoxycarbonyl)amino]-2-(3-cyanophenyl)propionate I-3d (400 mg, 47.08%) as a white solid. LCMS:m / z(ESI),[M+H- t Bu] + =248.95. 1 HNMR(400MHz,DMSO-d6)δ1.31(9H,s),3.33–3.42(1H,m),3.46–3.58(1H,m),3.6 3(3H,s),3.96(1H,t),6.96(1H,t),7.47–7.65(2H,m),7.73(1H,d),7.77(1H,d).

[0241] Step 4: At room temperature and under air atmosphere, a mixture of LiOH·H₂O (86.17 mg, 2.05 mmol, 2.5 equivalents) and nitrile I-3d (250 mg, 0.82 mmol, 1 equivalent) in THF (5 mL) and H₂O (1 mL) was stirred for 2 hours. The resulting mixture was concentrated under reduced pressure to give 3-[(tert-butoxycarbonyl)amino]-2-(3-cyanophenyl)propionic acid I-3 (200 mg, 83.87%) as a white solid. LCMS: m / z (ESI), [M+H] + =291.00.

[0242] Synthesis of intermediate I-4

[0243]

[0244] Step 1: At room temperature and under a nitrogen atmosphere, EDCI (2227.74 mg, 11.620 mmol, 2 equivalents) was added in portions to a stirred mixture of 2-(4-bromophenyl)-3-[(tert-butoxycarbonyl)amino]propionic acid (2.0 g, 5.81 mmol, 1 equivalent) and DMAP (141.97 mg, 1.16 mmol, 0.2 equivalents) in DCM (10 mL) and methanol (1.0 mL, 24.70 mmol, 4.25 equivalents). The mixture was stirred overnight at room temperature and under a nitrogen atmosphere. After the reaction, the mixture was diluted with water (40 mL). The mixture was extracted with ethyl acetate (3 x 30 mL). The combined organic layers were washed with water (3 x 20 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using PE / EA (4:1, v / v) elution to give methyl 2-(4-bromophenyl)-3-[(tert-butoxycarbonyl)amino]propionate I-4a (1.58 g, 75.91%) as a white solid. LCMS: m / z (ESI), [Mt-Bu] + =301.80.

[0245] Step 2: At room temperature and under a nitrogen atmosphere, Pd(PPh3)4 (477.42 mg, 0.41 mmol, 0.1 equivalent) was added in portions to a stirred mixture of ester I-4a (1.48 g, 4.13 mmol, 1 equivalent) and Zn(CN)2 (494.81 mg, 4.21 mmol, 1.02 equivalent) in DMF (4 mL). The resulting mixture was stirred at 80 °C under a nitrogen atmosphere for 3 hours. After cooling to room temperature, the mixture was diluted with water (25 mL). The mixture was extracted with ethyl acetate (3 x 30 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The crude product was diluted with DCM (3 mL). The residue was purified by preparative TLC with PE / EA (3:1, v / v) to give methyl 3-[(tert-butoxycarbonyl)amino]-2-(4-cyanophenyl)propionate I-4b (1.1 g, 87.48%) as a yellow oil. LCMS: m / z (ESI), [Mt-Bu] + =249.30.

[0246] Step 3: At 0°C under a nitrogen atmosphere, NaBH4 (298.32 mg, 7.89 mmol, 8 equivalents) was added in portions to a stirred mixture of nitrile I-4b (300 mg, 0.99 mmol, 1 equivalent) and (Boc)2O (430.26 mg, 1.97 mmol, 2 equivalents) in methanol (10 mL). The resulting mixture was stirred at 25°C under a nitrogen atmosphere for 3 hours. The reaction was quenched with ice water (30 mL) at 0°C. The resulting mixture was filtered, and the filter cake was washed with methanol (3 x 20 mL). The filtrate was concentrated under reduced pressure. The resulting mixture was extracted with ethyl acetate (3 x 30 mL). The combined organic layers were washed with water (3 x 10 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC with PE / EA (5:1, v / v) to obtain methyl 3-[(tert-butoxycarbonyl)amino]-2-(4-{[(tert-butoxycarbonyl)amino]methyl}phenyl)propionate I-4c (170 mg, 42.22%) as a yellow oil. LCMS: m / z (ESI), [M+Na) + =431.10.

[0247] Step 4: At 25°C under a nitrogen atmosphere, water (2.0 mL) containing LiOH·H₂O (65.74 mg, 1.57 mmol, 4.0 equivalent) was added dropwise to a stirred mixture of ester I-4c (160 mg, 0.39 mmol, 1 equivalent) in methanol (4.0 mL). The resulting mixture was stirred at 60°C under a nitrogen atmosphere for 3 hours. After cooling to room temperature, the mixture was acidified to pH 4 with HCl (1 M). The resulting mixture was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water, 10% to 50% gradient, 10 min; detector, UV 254 nm, 30% hold for 20 min. After evaporation of the solvent, 3-[(tert-butoxycarbonyl)amino]-2-(4-{[(tert-butoxycarbonyl)amino]methyl}phenyl)propionic acid I-4 (90 mg, 58.25%) was obtained as a white solid. LCMS: m / z (ESI), [MH] - =393.15.

[0248] Synthesis of intermediate I-5

[0249]

[0250] Step 1: At room temperature, add 100 mL of DMSO containing 3-iodonitrobenzene I-5a (12.56 g, 50.46 mmol, 1 equivalent), K₂CO₃ (20.92 g, 151.38 mmol, 3 equivalents), CuI (9.61 g, 50.46 mmol, 1 equivalent), and methyl cyanoacetate (5 g, 50.46 mmol, 1 equivalent) to a 20 mL vial. Stir the resulting mixture at 120 °C under a nitrogen atmosphere for 3 hours. Dilute the resulting mixture with EA (200 mL) and wash with 1 x 200 mL of saturated NaHCO₃ (aqueous solution), water (2 x 200 mL), and saturated brine (1 x 200 mL). Concentrate the organic layer under reduced pressure. Purify the residue by silica gel column chromatography using PE / EA (1:1, v / v) elution. The residue was purified by silica gel column chromatography using PE / EA (25:1) elution to obtain methyl 2-cyano-2-(3-nitrophenyl)acetate I-5b (3.0 g, 27.00%), which was a red oil. 1 H NMR (DMSO-d6, 400MHz) δ3.77(3H,s),6.01(1H,s),7.79(1H,t),7.93(1H,d),8.30(1H,dd),8.34(1H,t).

[0251] Step 2: At 0°C, NaBH4 (604.77 mg, 15.98 mmol, 8 equivalents) was added to a mixture of ester I-5b (440 mg, 2.00 mmol, 1 equivalent), CoCl2·6H2O (1.24 g, 5.20 mmol, 2.6 equivalents), and Boc2O (1.74 g, 7.99 mmol, 4 equivalents) in methanol (10 mL). The resulting mixture was stirred for 1 hour at room temperature. The mixture was quenched with water (50 mL), filtered, and the filter cake was washed with DCM (3 x 10 mL). The filtrate was concentrated under reduced pressure. The resulting mixture was extracted with DCM (3 x 50 mL). The organic layer was concentrated under reduced pressure. The residue was purified by preparative TLC with PE / EA (2:1, v / v) to obtain methyl 3-[(tert-butoxycarbonyl)amino]-2-{3-[(tert-butoxycarbonyl)amino]phenyl}propionate I-5c (300 mg, 38.06%) as a yellow oil. LCMS: m / z (ESI), [M+H + =395.20. 1 H NMR(CD3OD,400MHz)δ1.43(9H,s),1.54(9H,s),3.35–3.42(1H,m),3.61(1H, dd),3.70(3H,s),3.85(1H,dd),6.94(1H,d),7.23(1H,t),7.27–7.43(2H,m).

[0252] Step 3: At room temperature, add 2.5 mL of THF containing ester I-5c (300 mg, 0.76 mmol, 1 equivalent) and LiOH (36.43 mg, 1.52 mmol, 2 equivalent) and 2.5 mL of water to a 50 mL round-bottom flask. Stir the resulting mixture at room temperature for 1.5 hours. Acidify the mixture to pH 5 with HCl (1 M). Purify the residue by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, methanol / water, 0% to 20% gradient, 30 min; detector, UV 254 nm. Concentrate the resulting mixture under vacuum to give 3-[(tert-butoxycarbonyl)amino]-2-{3-[(tert-butoxycarbonyl)amino]phenyl}propionic acid I-5 (200 mg, 69.12%) as a yellow oil. 1 H NMR (CD3OD, 400MHz) δ1.42(9H,s),1.53(9H,s),3.40(1H,dd),3.48(1H,dd),3.61(1H,dd),7.03(1H,d),7.17(1H,d),7.21(1H,dd),7.42(1H,d).

[0253] Synthesis of intermediate I-6

[0254]

[0255] Step 1: At room temperature, ethyl cyanoacetate (2.26 g, 19.98 mmol, 1 equivalent), oxepane-3-one I-6a (2 g, 19.98 mmol, 1 equivalent), etidin (5.06 g, 19.98 mmol, 1 equivalent), L-proline (0.46 g, 4.00 mmol, 0.2 equivalent), and methanol (100 mL) were added to a 250 mL round-bottom flask. The mixture was stirred for 15 hours at 25 °C under a nitrogen atmosphere. The mixture was then concentrated under reduced pressure. The residue was purified by silica gel column chromatography using PE / EA (2:1, v / v) elution to give ethyl 2-cyano-2-(oxepane-3-yl)acetate I-6b (3.2 g, 81.22%) as a yellow solid. 1 H NMR(CDCl3,400MHz)δ1.35(3H,td),1.47–1.79(3H,m),1.85–2.04(1H,m),2.29–2.40(1H,m),3.36–3.53(3H,m),3.88(1H,ddt),4.26–4.34(2H,m).

[0256] Step 2: At 0°C, add 80 mL of methanol containing ester I-6b (3.1 g, 15.72 mmol, 1 equivalent), Boc₂O (6.86 g, 31.43 mmol, 2 equivalents), and CoCl₂·6H₂O (7.48 g, 31.43 mmol, 2 equivalents) to a 250 mL round-bottom flask. At 0°C, add NaBH₄ (2.97 g, 78.59 mmol, 5 equivalents) to the mixture in portions. Stir the resulting mixture for 12 hours at room temperature under a nitrogen atmosphere. Quench the reaction by adding water (200 mL) at 0°C. Filter the resulting mixture and wash the filter cake with DCM (6 x 50 mL). Extract the aqueous layer with CH₂Cl₂ (3 x 200 mL). Concentrate the resulting mixture under reduced pressure. The residue was purified by silica gel column chromatography using PE / EA (2:1, v / v) elution to obtain ethyl 3-[(tert-butoxycarbonyl)amino]-2-(oxacyclohexane-3-yl)propionate I-6c (3 g, 63.33%) as a yellow oil. LCMS: m / z (ESI), [M+H- t Bu] + =246.05. 1H NMR(CDCl3,400MHz)δ1.26–1.31(3H,m),1.44(9H,s),1.63(2H,dq),1.78(1H,d),1. 93(2H,dh),2.51(1H,dtd),3.15–3.48(4H,m),3.77–3.96(2H,m),4.13–4.23(2H,m).

[0257] Step 3: At room temperature, ester I-6c (1 g, 3.32 mmol, 1 equivalent) and LiOH·H2O (158.93 mg, 6.64 mmol, 2 equivalents), H2O (4 mL), and methanol (8 mL) were added to a 50 mL round-bottom flask. The mixture was stirred at 60 °C under a nitrogen atmosphere for 15 hours. The mixture was acidified to pH 5 with HCl (1 M). The mixture was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, methanol / water, 0% to 30% gradient, 40 min; detector, UV 220 nm. The mixture was concentrated under reduced pressure. The resulting solid was dried by lyophilization. This yielded 3-[(tert-butoxycarbonyl)amino]-2-(oxecyclohexane-3-yl)propionic acid I-6 (650 mg, 71.67%) as a white solid. LCMS:m / z(ESI),[M+H- t Bu] + =218.15. 1 H NMR(CDCl3,400MHz)δ1.48(9H,d),1.65(2H),1.74–2.06(2H,m),2.51(1H,s ),3.14(1H,s),3.20–3.54(4H,m),3.89(2H,dt),6.77(1H,s),10.29(1H,s).

[0258] Synthesis of intermediate I-7

[0259]

[0260] Step 1: At 0°C, 2-(pyridin-3-yl)acetonitrile I-7a (6 g, 50.79 mmol, 1 equivalent) and THF (40 mL, 493.71 mmol), sodium hydride (2.23 g, 55.87 mmol, 1.1 equivalent, 60%) were added to a 250 mL round-bottom flask. The mixture was stirred for 1 hour at room temperature, and then dimethyl carbonate (36.60 g, 406.30 mmol, 8 equivalents) was added. The mixture was stirred for 4 hours at room temperature. The mixture was extracted with CH2Cl2 / methanol = 20:1 (3 x 30 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using PE / EA (1:1, v / v) elution to obtain methyl 2-cyano-2-(pyridin-3-yl)acetate I-7b (7.4 g, 54.83%) as a brownish-yellow solid. LCMS: m / z (ESI), [M+H] + =177.25.

[0261] Step 2: Add 40 mL of methanol containing ester I-7b (2 g, 11.35 mmol, 1 equivalent), CoCl2·6H2O (8.10 g, 34.06 mmol, 3 equivalents), and Boc2O (4.96 g, 22.70 mmol, 2 equivalents) to a 100 mL round-bottom flask. Add NaBH4 (3.44 g, 90.82 mmol, 8 equivalents) at 0 °C. Stir the resulting mixture at 0 °C for 2 hours. Quench the reaction with water at room temperature. Extract the resulting mixture with CH2Cl2 (3 x 30 mL) and dry it over anhydrous Na2SO4. After filtration, concentrate the filtrate under reduced pressure. The residue was purified by preparative TLC using DCM / EA (1:1, v / v) to give methyl 3-[(tert-butoxycarbonyl)amino]-2-(pyridin-3-yl)propionate I-7c (310 mg, 8.14%) as a yellow solid. LCMS: m / z (ESI), [M+H] + =281.15.

[0262] Step 3: At room temperature, ester I-7c (300 mg, 1.07 mmol, 1 equivalent), LiOH (51.26 mg, 2.14 mmol, 2 equivalents), THF (5 mL), and water (5 mL) were added to a 50 mL round-bottom flask. The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water, 10% to 50% gradient, 40 min; detector, UV 220 nm. This yielded 3-[(tert-butoxycarbonyl)amino]-2-(pyridin-3-yl)propionic acid I-7 (140 mg, 41.16%) as a yellow solid. LCMS: m / z (ESI), [M+H)+ =267.05.

[0263] Synthesis of intermediate I-8

[0264]

[0265] Step 1. At room temperature, add atropine I-8a (8 g, 54.00 mmol, 1 equivalent) and ethanol (220 mL), H2SO4 (0.5 mL, 9.38 mmol, 0.17 equivalent) to a 250 mL round-bottom flask. Stir the resulting mixture overnight at 80 °C. After cooling to room temperature, neutralize the mixture to pH 8 with saturated NaHCO3 (aqueous solution). Dilute the resulting mixture with water (70 mL). Extract the aqueous layer with ethyl acetate (3 x 100 mL). Concentrate the resulting mixture under reduced pressure. Purify the residue by silica gel column chromatography using PE / EA (10:1, v / v) elution to give ethyl 2-phenylprop-2-enoate I-8b (7.5 g, 78.82%) as a colorless liquid. 1 H NMR (400MHz, DMSO-d6) δ1.3(t,3H),4.2(q,2H),6.0(d,1H),6.2(d,1H),7.2–7.5(m,5H).

[0266] Step 2: At room temperature and under a nitrogen atmosphere, a solution of ester I-8b (250 mg, 1.50 mmol, 1 equivalent) and 1-methylpiperazine (568.42 mg, 5.68 mmol, 4 equivalents) in DMF (3 mL) was stirred for 1 hour. The residue was dissolved in PE (20 mL). The combined organic layers were washed with water (3 x 10 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure to give ethyl 3-(4-methylpiperazin-1-yl)-2-phenylpropionate I-8c (330 mg, 84.16%) as a grayish-white solid. LCMS: m / z (ESI), [M+H] + =277.25.

[0267] Step 3: At room temperature and under air, LiOH (173.31 mg, 7.24 mmol, 4 equivalents) was added dropwise to a stirred solution of piperazine I-8c (500 mg, 1.81 mmol, 1 equivalent) in methanol (10 mL) and H2O (5 mL). The resulting mixture was stirred for 3 hours at room temperature and under air. The resulting mixture was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water, 20% to 30% gradient, 10 min; detector, UV 254 nm, to give 3-(4-methylpiperazin-1-yl)-2-phenylpropionic acid I-8 (220 mg, 48.97%) as a white solid. LCMS: m / z (ESI), [M+H] + =249.15.

[0268] Synthesis of intermediate I-9

[0269]

[0270] Step 1: At 0°C and under air, t-BuOK (1.37 g, 12.18 mmol, 2 equivalents) was added in portions to a stirred mixture of ethyl phenylacetate I-9a (1000 mg, 6.09 mmol, 1 equivalent) and tert-butyl 4-iodopiperidine-1-carboxylate I-9b (2.84 g, 9.14 mmol, 1.5 equivalents) in 10 mL of DMF. The resulting mixture was stirred at 25°C and under air for 16 hours. The reaction was quenched with saturated NH4Cl (aqueous solution) at 0°C. The resulting mixture was extracted with ethyl acetate (3 x 50 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography under the following conditions: column, silica gel; mobile phase, MeCN / water, 50% to 90% gradient, 10 min; detector, UV 254 nm, to obtain tert-butyl 4-(2-ethoxy-2-oxo-1-phenylethyl)piperidine-1-carboxylate (550 mg, 25.99%) as a yellow solid. 1 HNMR(400MHz,DMSO-d6)δ1.13(4H,t),1.37(9H,s),1.67(1H,d),2.01-2.1 6(1H,m),3.37(4H,d),3.83(1H,d),3.90-4.17(3H,m),7.25-7.37(5H,m).

[0271] Step 2: Add TFA (2 mL) to a stirred solution of tert-butyl 4-(2-ethoxy-2-oxo-1-phenylethyl)piperidine-1-carboxylate (500 mg, 1.44 mmol, 1 equivalent) in DCM (6 mL). Stir the resulting mixture for 2 hours at room temperature under air atmosphere. Concentrate the resulting mixture under vacuum to give ethyl 2-phenyl-2-(piperidin-4-yl)acetate I-9c (340 mg, 95.52%) as a yellow oil. LCMS: m / z (ESI), [M+H] + =248.15.

[0272] Step 3: DIEA (914.47 mg, 7.08 mmol, 5 equivalents) was added to a stirred mixture of ester I-9c (350 mg, 1.42 mmol, 1 equivalent) and formaldehyde (424.89 mg, 14.15 mmol, 10 equivalents) in a DCM (10 mL). The resulting mixture was stirred for 0.5 h at room temperature under air. NaBH(OAc)3 (899.73 mg, 4.25 mmol, 3 equivalents) was added to the mixture. The resulting mixture was stirred for another 2 h at room temperature. The reaction was quenched by adding water (4 mL) at 0 °C. The resulting mixture was extracted with ethyl acetate (3 x 30 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. This yielded ethyl 2-(1-methylpiperidin-4-yl)-2-phenylacetate (350 mg, 94.63%) as a yellow oil. LCMS:m / z(ESI),[M+H] + =262.10.

[0273] Step 4: LiOH·H2O (67.43 mg, 1.61 mmol, 1.2 equivalents) was added to a stirred solution of ethyl 2-(1-methylpiperidin-4-yl)-2-phenylacetic acid (350 mg, 1.34 mmol, 1 equivalent) in THF (4 mL) and H2O (1 mL). The resulting mixture was stirred at 70 °C under a nitrogen atmosphere for 16 hours. The mixture was acidified to pH 6 with HCl (1 M). The resulting mixture was concentrated under vacuum. The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, methanol / water, 10% to 50% gradient, 10 min; detector, UV 254 nm, to give (1-methylpiperidin-4-yl)(phenyl)acetic acid I-9 (240 mg, 76.82%) as a white solid. LCMS: m / z (ESI), [M+H) + =234.15. 1H NMR(400MHz,DMSO-d6)δ7.41-7.19(5H,m),3.18(1H,d),2.85(1H,d),2.71(1H,d),2 .19(3H,t),1.99(1H,s),1.86(2H,d),1.76(1H,d),1.31(1H,q),1.14-0.99(2H,m).

[0274] Synthesis of intermediate I-10

[0275]

[0276] Step 1: Under a nitrogen atmosphere, at 70 °C, a mixture of methyl 2-bromo-2-phenylacetate I-10a (10 g, 43.65 mmol, 1 equivalent), 1-methylpiperazine (13.12 g, 130.96 mmol, 3 equivalents), and triethylamine (4.42 g, 43.65 mmol, 1 equivalent) in 100 mL of THF was stirred for 4 hours. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using CH2Cl2 / methanol (15:1, v / v) elution to give methyl 2-(4-methylpiperazin-1-yl)-2-phenylacetate I-10b (6.3 g, 58.12%) as a brown oil. LCMS: m / z (ESI), [M+H] + =249.15.

[0277] Step 2: At room temperature, add 16 mL of methanol and 8 mL of water containing ester I-10b (2.7 g, 10.87 mmol, 1 equivalent) and LiOH·H₂O (0.91 g, 21.75 mmol, 2.0 equivalent) to a 50 mL round-bottom flask. Stir the resulting mixture for 5 hours at room temperature under a nitrogen atmosphere. Acidify the mixture to pH 6 with HCl (1 M). Purify the residue by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, methanol / water, 0% to 5% gradient, 30 min; detector, UV 220 nm, to give (4-methylpiperazin-1-yl)(phenyl)acetic acid I-10 (1.6 g, 62.81%) as a white solid. LCMS: m / z (ESI), [M+H] + =235.15.

[0278] Synthesis of intermediate I-11

[0279]

[0280] Step 1: At 0°C and under air, NaBH4 (16.20 g, 428.12 mmol, 5 equivalents) was added in portions to a stirred solution of methyl 3-(cyanomethyl)benzoate I-11a (15 g, 85.62 mmol, 1 equivalent) in methanol (250 mL). The resulting mixture was stirred overnight at 70°C and under air. The mixture was cooled to room temperature. The resulting mixture was concentrated under reduced pressure. The reaction was quenched by adding water (100 mL) at room temperature. The resulting mixture was extracted with ethyl acetate (3 x 50 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using PE / EA (1:10, v / v) elution to give 2-[3-(hydroxymethyl)phenyl]acetonitrile (11.14 g, 88.40%) as a yellow oil. 1 H NMR (DMSO-d6, 400MHz) δ4.10(2H,s),4.52(2H,d),5.28(1H,t),7.21(1H,d),7.27(1H,d),7.32–7.39(2H,m).

[0281] Step 2: At room temperature and under air, DMAP (0.92 g, 7.569 mmol, 1 equivalent) and triethylamine (22.98 g, 227.073 mmol, 3 equivalent) were added dropwise to a stirred solution of 2-[3-(hydroxymethyl)phenyl]acetonitrile (11.14 g, 75.69 mmol, 1 equivalent) and TBSCl (17.11 g, 113.54 mmol, 1.5 equivalent) in 100 mL of DCM. The resulting mixture was stirred overnight at room temperature and under air. The reaction was quenched by adding water (100 mL) at room temperature. The mixture was extracted with CH2Cl2 (3 x 100 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using PE / EA (1:1, v / v) elution to obtain 2-(3-{[(tert-butyldimethylsilyl)oxy]methyl}phenyl)acetonitrile I-11b (19.5 g, 98.54%) as a colorless oil. 1 H NMR(DMSO-d6,400MHz)δ0.09(6H,s),0.92(9H,s),4.04(2H,s),4.73(2H,s),7.24(2H,dd),7.35(2H,dd).

[0282] Step 3: At 0°C and under air, sodium hydride (3.58 g, 149.17 mmol, 2 equivalents) was added dropwise to a stirred solution of nitrile I-11b (19.5 g, 74.59 mmol, 1 equivalent) in 100 mL of THF. The resulting mixture was stirred for 1 hour at room temperature under air. Dimethyl carbonate (53.75 g, 596.70 mmol, 8 equivalents) was added dropwise to the mixture at room temperature. The resulting mixture was stirred for another 3 hours at room temperature. The reaction was quenched by adding water / ice (100 mL) at room temperature. The resulting mixture was extracted with ethyl acetate (3 x 100 mL). The combined organic layers were dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using PE / EA (1:1, v / v) elution to obtain methyl 2-(3-{[(tert-butyldimethylsilyl)oxy]methyl}phenyl)-2-cyanoacetate (10.1 g, 42.39%), which was a yellow oil. 1 H NMR (CD3OD, 400MHz) δ0.13(6H,d),0.97(9H,s),3.78(3H,s),4.79(2H,d),7.22–7.46(3H,m),7.50(1H,dq).

[0283] Step 4: At room temperature and under air atmosphere, add Boc2O (10.25 g, 46.95 mmol, 3 equivalents) to a stirred mixture of methyl 2-(3-{[(tert-butyldimethylsilyl)oxy]methyl}phenyl)-2-cyanoacetate (5 g, 15.65 mmol, 1 equivalent) and CoCl2·6H2O (11.17 g, 46.95 mmol, 3 equivalents) in methanol (100 mL). Add NaBH4 (4.74 g, 125.21 mmol, 8 equivalents) in portions to the mixture at 0 °C. Stir the resulting mixture at room temperature for another 3 hours. Quench the reaction by adding water (100 mL) at room temperature. Extract the resulting mixture with CH2Cl2 (3 x 50 mL). Dry the combined organic layers with anhydrous Na2SO4. After filtration, concentrate the filtrate under reduced pressure. The residue was purified by silica gel column chromatography using PE / EA (1:1, v / v) elution to obtain methyl 3-[(tert-butoxycarbonyl)amino]-2-(3-{[(tert-butyldimethylsilyl)oxy]methyl}phenyl)propionate I-11c (2282 mg, 34.42%), a yellow oil. LCMS: m / z (ESI), [M+H-Boc] + =324.15. 1H NMR(CD3OD,400MHz)δ0.12(6H,s),0.97(9H,d),1.43(9H,d),3.39(1H,dd),3.61( 1H,dd),3.69(3H,s),3.90(1H,dd),4.75(2H,d),7.19(1H,d),7.23–7.34(3H,m).

[0284] Step 5: At room temperature and under air, TBAF (370.32 mg, 1.42 mmol, 3 equivalents) was added to a stirred solution of ester I-11c (200 mg, 0.472 mmol, 1 equivalent) in THF (5 mL, 61.71 mmol). The resulting mixture was stirred for 3 hours at room temperature and under air. The mixture was then concentrated under reduced pressure. The residue was purified by preparative TLC with CH2Cl2 / methanol (40:1, v / v) to give methyl 3-[(tert-butoxycarbonyl)amino]-2-[3-(hydroxymethyl)phenyl]propionate (97 mg, 66.41%) as a white solid. LCMS: m / z (ESI), [M+H] + =310.20. 1 HNMR(DMSO-d6,400MHz)δ1.37(9H,d),3.23(1H,dt),3.50(1H,ddd),3.60(3H,s),3.86(1H,dt ),4.48(2H,t),5.21(1H,t),6.85–7.08(1H,m),7.09–7.15(1H,m),7.23(2H,d),7.30(1H,t).

[0285] Step 6: At room temperature and under air, N-methylcarbamoyl chloride (108.82 mg, 1.16 mmol, 1.2 equivalents) and Et3N (294.39 mg, 2.91 mmol, 3 equivalents) were added dropwise to a stirred solution of methyl 3-[(tert-butoxycarbonyl)amino]-2-[3-(hydroxymethyl)phenyl]propionate (300 mg, 0.97 mmol, 1 equivalent) in DCM (6 mL, 94.38 mmol, 97.33 equivalents). The resulting mixture was stirred overnight at room temperature and under air. The reaction was quenched by adding water (10 mL) at room temperature. The resulting mixture was extracted with CH2Cl2 (3 x 10 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water, 100% to 100% gradient, 10 min; detector, UV 254 nm, to obtain methyl 3-[(tert-butoxycarbonyl)amino]-2-(3-{[(methylcarbamoyl)oxy]methyl}-phenyl)propionate I-11d (90 mg, 25.33%) as a colorless oil. LCMS: m / z (ESI), [M+H] + =367.15.

[0286] Step 7: At room temperature and under air, LiOH (70.59 mg, 2.95 mmol, 4 equivalents) was added to a stirred solution of ester I-11d (270 mg, 0.74 mmol, 1 equivalent) in methanol (8 mL) and H2O (2 mL). The resulting mixture was stirred for 2 hours at room temperature and under air. The resulting mixture was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water, 50% to 60% gradient, 10 min; detector, UV 254 nm, to give 3-[(tert-butoxycarbonyl)amino]-2-(3-{[(methylcarbamoyl)oxy]methyl}phenyl)propionic acid I-11 (90 mg, 34.66%) as a white solid. LCMS: m / z (ESI), [M+H] + =353.20.

[0287] Synthesis of intermediate I-12

[0288]

[0289] Step 1: At 25°C under a nitrogen atmosphere, water (10 mL) containing NH₄Cl (4.57 g, 85.39 mmol, 1 equivalent) was added dropwise to a stirred mixture of ethyl 2-cyano-2-(4-nitrophenyl)-2-cyanoethyl I-12a (2.0 g, 8.539 mmol, 1 equivalent) and zinc (4.47 g, 68.31 mmol, 8 equivalent) in 40 mL of THF. The mixture was stirred at room temperature under a nitrogen atmosphere for 48 hours. After the reaction, the mixture was filtered, and the filter cake was washed with methanol (3 x 20 mL). The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using CH₂Cl₂ / methanol (60:1, v / v) elution to give ethyl 2-(4-aminophenyl)-2-cyanoethyl I-12b (1.2 g, 68.81%) as a yellow solid. LCMS:m / z(ESI),[M+H] + =205.15.

[0290] Step 2: At 0°C and under a nitrogen atmosphere, triethylamine (1.64 g, 16.16 mmol, 3 equivalents) was added dropwise to a stirred mixture of nitrile I-12b (1.1 g, 5.39 mmol, 1 equivalent) and triethylamine (1.64 g, 16.16 mmol, 3 equivalents) in DCM (30 mL). The resulting mixture was stirred for 2 hours at room temperature under a nitrogen atmosphere. After the reaction, the resulting mixture was diluted with water (80 mL). The mixture was extracted with ethyl acetate (3 x 60 mL). The combined organic layers were washed with saturated brine (3 x 100 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was dissolved in DCM (5 mL). The residue was purified by preparative TLC with PE / EA (1:1, v / v) to give ethyl 2-cyano-2-(4-acetamidophenyl)acetate I-12c (1.2 g, 90.47%) as a yellow oil. LCMS: m / z (ESI), [M+H] + =247.15. 1 H NMR (DMSO-d6, 400MHz) δ1.18(3H,t),2.06(3H,s),4.18(2H,qq),5.56(1H,s),7.34(2H,m),7.65(2H,m),10.08(1H,s).

[0291] Step 3: At 0°C and under air, (Boc)₂O (1.95 g, 8.93 mmol, 1 equivalent) and NaBH₄ (0.68 g, 17.87 mmol, 4 equivalent) were added in portions to a stirred mixture of ester I-12c (1.1 g, 4.47 mmol, 1 equivalent) and CoCl₂·6H₂O (2.13 g, 8.93 mmol, 2 equivalent) in methanol (40 mL). The resulting mixture was stirred overnight at room temperature under air. After the reaction, the mixture was filtered, and the filter cake was washed with water (3 x 30 mL). The mixture was diluted with water (50 mL). The mixture was extracted with ethyl acetate (3 x 40 mL). The combined organic layers were dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC using CH₂Cl₂ / methanol (5:1, v / v) to give ethyl 3-[(tert-butoxycarbonyl)amino]-2-(4-acetamidophenyl)propionate I-12d (900 mg, 57.50%) as a yellow oil. LCMS: m / z (ESI), [M-Boc] + =251.15.

[0292] Step 4: At room temperature, under a nitrogen atmosphere, water (1.0 mL) containing LiOH (194.51 mg, 8.12 mmol, 4 equivalents) was added dropwise to a stirred mixture of ester I-12d (500 mg, 2.03 mmol, 1 equivalent) in methanol (4.0 mL). The resulting mixture was stirred overnight at 60 °C under a nitrogen atmosphere. After cooling to room temperature, the resulting mixture was concentrated under vacuum. The resulting mixture was diluted with methanol (3.0 mL). The mixture was acidified to pH 5 with HCl (1 M). The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water, 10% to 50% gradient, 30 min; detector, UV 254 nm, 16% hold for 10 min. After lyophilization, 2-(4-acetamidophenyl)-3-((tert-butoxycarbonyl)amino)propionic acid I-12 (260 mg, 36.54%) was obtained as a white solid. LCMS: m / z (ESI), [Mt-Bu] + =267.05.

[0293] Synthesis of intermediate I-13

[0294]

[0295] Step 1: At 100°C under a nitrogen atmosphere, Pd(OAc)₂ (0.28 g, 1.27 mmol, 0.2 equivalent), K₃PO₄ (2.69 g, 12.65 mmol, 2.0 equivalent), and XantPhos (1.10 g, 1.90 mmol, 0.3 equivalent) were added in portions to a mixture of N-[2-(3-bromophenyl)-2-hydroxyethyl]carbamate tert-butyl I-13a (2.0 g, 6.33 mmol, 1 equivalent) and (methylphosphoryl)methane (0.59 g, 7.59 mmol, 1.2 equivalent) in dioxane (25 mL). The mixture was stirred overnight. The mixture was cooled to room temperature. The reaction was quenched with water at room temperature. The resulting mixture was extracted with ethyl acetate (3 x 30 mL). The combined organic layers were dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water (0.1% FA), 0% to 100% gradient, 10 min; detector, UV 254 nm. This yielded N-{2-[3-(dimethylphosphoryl)phenyl]-2-hydroxyethyl} tert-butyl carbamate I-13b (1.3 g, 65.59%) as a yellow oil. 1 H NMR (400MHz, DMSO-d6) δ1.35(9H,s),1.64(6H,d),3.11(2H,m),4.65(1H,q),5.48(1H,d),6.78(1H,t),7.48(2H,dd),7.64(1H,ddd),7.72(1H,d).

[0296] Step 2: At 0°C and under a nitrogen atmosphere, a mixture of 169.05 mg phthalimide (1.15 mmol, 1.2 equivalents), PPh3 (753.39 mg, 2.87 mmol, 3 equivalents) in THF (1 mL) was treated with DIAD (580.81 mg, 2.87 mmol, 3 equivalents) for 0.5 h. Subsequently, at 0°C, 2 mL of THF containing alcohol I-13b (300 mg, 0.96 mmol, 1 equivalent) was added dropwise for 1 h. The reaction was quenched with water at room temperature. The resulting mixture was extracted with CH2Cl2 (3 x 20 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC with CH2Cl2 / methanol (10:1, v / v) to give N-{2-[3-(dimethylphosphoryl)phenyl]-2-(1,3-dioxoisoindol-2-yl)ethyl} tert-butyl carbamate I-13c (270 mg, 63.74%) as a white solid. 1H NMR (400MHz, DMSO-d6) δ1.27(9H,s),1.64(6H,dd),3.89(2H,m),5.43(1H,dd),7.27(1H,t),7.54(2H,m),7.72(2H,m),7.87(4H,m).

[0297] Step 3: At room temperature and under a nitrogen atmosphere, a solution of o-benzoimide I-13c (240 mg, 0.54 mmol, 1 equivalent) and hydrazine (216.97 mg, 4.34 mmol, 8 equivalents) in methanol (5 mL) was stirred for 4 hours. The resulting mixture was filtered, and the filter cake was washed with methanol (3 x 5 mL). The filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water, 10% to 50% gradient, 10 min; detector, UV 254 nm. This yielded N-{2-amino-2-[3-(dimethylphosphoryl)phenyl]ethyl}-tert-butyl carbamate I-13 (82 mg, 48.40%) as a colorless oil. 1 H NMR (400MHz, DMSO-d6) δ1.35(9H,s),1.63(6H,d),3.09(3H,m),6.84(1H,t),7.54(4H,m).

[0298] Synthesis of intermediate I-14

[0299]

[0300] Step 1: At room temperature and under a nitrogen atmosphere, Zn(CN)₂ (688.59 mg, 5.86 mmol, 1.01 equivalent) was added in portions to a stirred solution of ester I-26c (2.08 g, 5.81 mmol, 1 equivalent) and Pd(PPh₃)₄ (67.10 mg, 0.058 mmol, 0.01 equivalent) in DMF (10 mL). The resulting mixture was stirred at 80 °C under a nitrogen atmosphere for 2.0 h. After cooling to room temperature, the mixture was diluted with water (60 mL). The mixture was extracted with ethyl acetate (3 x 40 mL). The combined organic layers were washed with water (3 x 10 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC with PE / EA (2:1, v / v) to give methyl 3-[(tert-butoxycarbonyl)amino]-2-(4-cyanophenyl)propionate I-14c (1.3 g, 73.57%) as a yellow solid.

[0301] Step 2: At room temperature, under a nitrogen atmosphere, water (2.0 mL) containing LiOH·H₂O (330.89 mg, 7.88 mmol, 4.0 equivalents) was added dropwise to a stirred solution of nitrile I-14c (600 mg, 1.97 mmol, 1 equivalent) in methanol (8.0 mL). The resulting mixture was stirred at 70 °C under a nitrogen atmosphere for 2.0 h. After cooling to room temperature, the resulting mixture was concentrated under vacuum. After evaporation, the residue was dissolved in methanol (2.0 mL) and alkalized to pH 9 with an aqueous NH₃ solution. The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water, 10% to 50% gradient, 10 min; detector, UV 254 nm, 18.2% hold for 20 min, to give 3-[(tert-butoxycarbonyl)amino]-2-(4-cyanophenyl)propionic acid I-14 (400 mg, 69.89%) as a white solid. LCMS: m / z (ESI), [M- t Bu] + =235.05.

[0302] Synthesis of intermediate I-15

[0303]

[0304] Step 1: At 0°C, (5-bromopyridin-2-yl)methanol I-15a (5 g, 26.59 mmol, 1 equivalent), TBSCl (4.81 g, 31.91 mmol, 1.2 equivalent), triethylamine (8.07 g, 79.78 mmol, 3 equivalent), DMAP (0.32 g, 2.66 mmol, 0.1 equivalent), and DCM (80 mL) were added to a 250 mL round-bottom flask. The mixture was stirred overnight at room temperature. The mixture was extracted with CH2Cl2 (3 x 30 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using EA / CH2Cl2 (3:1, v / v) elution to give 5-bromo-2-{[(tert-butyldimethylsilyl)oxy]methyl}pyridine (6.33 g, 78.23%) as a yellow oil. LCMS:m / z(ESI),[M+H] + =304.05. 1 H NMR (CD3OD, 400MHz) δ0.15(6H,s),0.93(9H,d),4.77(2H,d),7.50(1H,m),8.01(1H,m),8.55(1H,m).

[0305] Step 2: At room temperature, 5-bromo-2-{[(tert-butyldimethylsilyl)oxy]methyl}pyridine (6.3 g, 20.84 mmol, 1 equivalent), methyl cyanoacetate (2.48 g, 25.01 mmol, 1.2 equivalent), Pd(OAc)₂ (0.47 g, 2.08 mmol, 0.1 equivalent), dppf (2.30 g, 4.17 mmol, 0.2 equivalent), t-BuOK (7.02 g, 62.52 mmol, 3 equivalent), and 1,4-dioxane (240 mL) were added to six 500 mL round-bottom flasks. The resulting mixture was stirred for 5 hours at 70 °C under a nitrogen atmosphere. The mixture was quenched with saturated brine (200 mL) and extracted with EA (2 x 200 mL). The combined organic layers were dried over anhydrous Na₂SO₄, filtered, and evaporated to obtain a crude solid. The residue was purified by silica gel column chromatography using PE / EA (1:1, v / v) elution to obtain methyl 2-(6-{[(tert-butyldimethylsilyl)oxy]methyl}pyridin-3-yl)-2-cyanoacetate I-15b (2.4 g, 33.38%) as a yellow oil. LCMS: m / z (ESI), [M+H) + =321.15. 1 H NMR (CD3OD, 400MHz) δ0.17(6H,s),0.99(9H,s),3.68(1H,d),3.83(3H,s),4.86(2H,s),7.66(1H,d),7.98(1H,dd),8.56(1H,d).

[0306] Step 3: At room temperature, add nitrile I-15b (2.24 g, 6.99 mmol, 1 equivalent), CoCl2·6H2O (2.49 g, 10.49 mmol, 1.5 equivalent), Boc2O (3.05 g, 13.98 mmol, 2.0 equivalent), and methanol (20 mL) to a 50 mL round-bottom flask. Add NaBH4 (1.06 g, 27.96 mmol, 4.0 equivalent) in portions at 0 °C. Stir the resulting mixture overnight at room temperature. Quench the reaction with water (100 mL) at room temperature. Filter the resulting mixture and wash the filter cake with CH2Cl2 (3 x 30 mL). Extract the filtrate with DCM (2 x 100 mL). Dry the combined organic layers with anhydrous Na2SO4. After filtration, concentrate the filtrate under reduced pressure. The residue was purified by preparative TLC using CH₂Cl₂ / methanol (17:1, v / v) to give methyl 3-[(tert-butoxycarbonyl)amino]-2-(6-{[(tert-butyldimethylsilyl)oxy]methyl}pyridin-3-yl)propionate (625 mg, 14.74%) as a brownish-yellow solid. LCMS: m / z (ESI), [M+H] +=425.30.

[0307] Step 4: At room temperature, methyl 3-[(tert-butoxycarbonyl)amino]-2-(6-{[(tert-butyldimethylsilyl)oxy]methyl}pyridin-3-yl)propionate (600 mg, 1.41 mmol, 1 equivalent), LiOH (67.69 mg, 2.83 mmol, 2 equivalent), water (5 mL), and THF (5 mL) were added to a 50 mL round-bottom flask. The mixture was stirred overnight at room temperature. The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, methanol / water, 0 to 10% gradient, 10 min; detector, UV 254 nm, to obtain 3-[(tert-butoxycarbonyl)amino]-2-[6-(hydroxymethyl)pyridin-3-yl]propionate I-15 (300 mg, 63.77%) as a light brown solid. LCMS: m / z (ESI), [M+H] + =297.10.

[0308] Synthesis of intermediate I-16

[0309]

[0310] Step 1: At 70 °C and under a nitrogen atmosphere, a mixture of 3-bromo-5-methylpyridine I-16a (5 g, 29.07 mmol, 1 equivalent), methyl 2-cyanoacetate (3.46 g, 34.88 mmol, 1.2 equivalent), Pd(OAc)₂ (0.65 g, 2.91 mmol, 0.1 equivalent), dppf (3.21 g, 5.81 mmol, 0.2 equivalent), and potassium 2-methylprop-2-ol (9.78 g, 87.20 mmol, 3 equivalent) in 1,4-dioxane (100 mL) was stirred for 2 hours. The reaction was monitored by LCMS. The mixture was acidified to pH 9 with saturated NH₄Cl (aqueous solution). The resulting mixture was stirred for 1 hour at room temperature. The mixture was extracted with ethyl acetate (3 x 300 mL). The combined organic layers were dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using CH2Cl2 / methanol (30:1, v / v) elution to give methyl 2-cyano-2-(5-methylpyridin-3-yl)acetate I-16b (710 mg, 12.84%) as a yellow solid. LCMS: m / z (ESI), [M+H] + =191.05.

[0311] Step 2: At 0°C and under a nitrogen atmosphere, NaBH4 (1272.92 mg, 33.65 mmol, 8 equivalents) was added in portions to a stirred mixture of nitrile I-16b (800 mg, 4.21 mmol, 1 equivalent) and Boc2O (3671.87 mg, 16.82 mmol, 4 equivalents) in methanol (50 mL). The resulting mixture was stirred for 1 hour at room temperature under a nitrogen atmosphere. The reaction was quenched with water at room temperature. The mixture was extracted with CH2Cl2 (2 x 100 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC using CH₂Cl₂ / methanol (32:1, v / v) to give methyl 3-[(tert-butoxycarbonyl)amino]-2-(5-methylpyridin-3-yl)propionate I-16c (310 mg, 43.66%) as a yellow solid. LCMS: m / z (ESI), [M+H] + =295.15.

[0312] Step 3: At room temperature and under a nitrogen atmosphere, a solution containing ester I-16c (300 mg, 1.02 mmol, 1 equivalent) of methanol (3 mL) and H2O containing LiOH·H2O (85.53 mg, 2.04 mmol, 2 equivalents) (1.5 mL) was stirred for 1 hour. The residue was acidified to pH 5 with HCl (1 M). The resulting mixture was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, methanol / water, 10% to 23% gradient, 30 min; detector, UV 220 nm, to give 3-[(tert-butoxycarbonyl)amino]-2-(5-methylpyridin-3-yl)propionic acid I-16 (140 mg, 49.00%) as a yellow solid. LCMS: m / z (ESI), [M+H] + =281.10.

[0313] Synthesis of intermediate I-17

[0314]

[0315] Step 1: At room temperature, add 70 mL of dioxane containing bromide I-17a (2.26 g, 22.80 mmol, 1.2 equivalents), dppf (1.05 g, 1.90 mmol, 0.1 equivalents), t-BuOK (6.40 g, 57.01 mmol, 3.0 equivalents), palladium acetate (0.21 g, 0.95 mmol, 0.05 equivalents), and methyl 2-cyano-2-methylacetate (2.58 g, 22.80 mmol, 1.2 equivalents) to a 250 mL round-bottom flask. Stir the resulting mixture at 70 °C under a nitrogen atmosphere for 3 hours. Quench the residue with saturated NH4Cl (100 mL). Filter the resulting mixture and extract the filtrate with ethyl acetate (2 x 150 mL). Dry the combined organic layers over anhydrous Na2SO4, filter, and concentrate under reduced pressure. The residue was purified by silica gel column chromatography using PE / EA (10:1, v / v) elution to obtain methyl 2-[3-(benzyloxy)phenyl]-2-cyanoacetate I-17b (1.8 g, 33.67%) as a yellow oil. 1 H NMR(DMSO-d6,400MHz)δ3.71(2H,s),3.73(3H,s),4.03(1H,s),5.13(2H,s ),7.01(1H,dt),7.03–7.12(2H,m),7.31–7.44(4H,m),7.45–7.50(2H,m).

[0316] Step 2: At 0°C, add 5 mL of methanol containing nitrile I-17b (2 g, 7.11 mmol, 1 equivalent), CoCl2·6H2O (4.40 g, 18.49 mmol, 2.6 equivalent), and Boc2O (6.21 g, 28.44 mmol, 4.0 equivalent) to a 40 mL vial. At 0°C, add NaBH4 (2.15 g, 56.88 mmol, 8.0 equivalent) to the above mixture. Stir the resulting mixture at room temperature for 2 hours. Quench the reaction with water at 0°C. Filter out the precipitate and wash with DCM (1 x 100 mL). Extract the resulting mixture with DCM (3 x 100 mL). Dry the combined organic layers with anhydrous Na2SO4, filter, and concentrate under reduced pressure. The residue was purified by preparative TLC with CH2Cl2 / methanol (10:1, v / v) to give methyl 2-[3-(benzyloxy)phenyl]-3-[(tert-butoxycarbonyl)amino]propionate I-17c (1.1 g, 40.14%) as a yellow solid. LCMS: m / z (ESI), [M+H- t Bu] + =330.10. 1HNMR(400MHz,DMSO-d6)δ1.35(9H,s),3.25(1H,dt),3.48(1H,ddd),3.59(3H,s),3.83(1H,t),5. 09(2H,d),6.83(1H,d),6.89(1H,t),6.94(2H,dd),7.26(1H,t),7.38–7.43(2H,m),7.46(2H,d).

[0317] Step 3: At room temperature, add 2 mL of THF containing ester I-17c (1.1 g, 2.85 mmol, 1 equivalent) and 2 mL of water containing LiOH·H₂O (239.49 mg, 5.71 mmol, 2 equivalents) to a 50 mL round-bottom flask. Stir the resulting mixture at room temperature for 1.5 hours. Acidify the mixture to pH 5 with HCl (aqueous solution). Purify the residue by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, water / methanol, 0% to 100% gradient, 30 min; detector, UV 254 nm. Concentrate the resulting mixture under vacuum to give 2-[3-(benzyloxy)phenyl]-3-[(tert-butoxycarbonyl)amino]propionic acid I-17 (500 mg, 47.17%) as a yellow solid. LCMS: m / z (ESI), [M+H- t Bu] + =316.10. 1 H NMR(400MHz,DMSO-d6)δ1.34(9H,s),3.23(1H,dt),3.44(1H,ddd),3.74(1H,t),5.08(2H,d),6 .85(1H,t),6.89–6.96(2H,m),7.25(1H,t),7.31–7.37(1H,m),7.37–7.43(2H,m),7.46(2H,d).

[0318] Synthesis of intermediate I-18

[0319]

[0320] Step 1: Under a nitrogen atmosphere, at 80 °C, a mixture of methyl 2-(pyridin-3-yl)acetate I-18a (4 g, 26.46 mmol, 1 equivalent), K₂CO₃ (10.97 g, 79.38 mmol, 3 equivalents), and formaldehyde (1.59 g, 52.92 mmol, 2 equivalents) in DMF (40 mL) was stirred for 2 hours. The mixture was cooled to room temperature. The reaction was quenched with water (60 mL) at room temperature. The resulting mixture was extracted with ethyl acetate (3 x 50 mL). The combined organic layers were washed with saturated sodium chloride solution (3 x 50 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using PE / EA (2:3, v / v) elution to give methyl 2-(pyridin-3-yl)prop-2-enoate I-18b (757 mg, 17.53%) as a yellow liquid. LCMS: m / z (ESI), [M+H] + =164.25. 1 H NMR (CDCl3, 400MHz) δ3.87(3H,s),6.02(1H,d),6.54(1H,d),7.34(1H,ddd),7.81(1H,ddd),8.60(1H,dd),8.68(1H,dd).

[0321] Step 2: At room temperature and under a nitrogen atmosphere, 1-methylpiperazine (2323.41 mg, 23.20 mmol, 5 equivalents) was added to a stirred mixture of enolate I-18b (757 mg, 4.64 mmol, 1 equivalent) in DMF (10 mL). The resulting mixture was stirred for 2 hours at room temperature and under a nitrogen atmosphere. The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water, 10% to 50% gradient, 10 min; detector, UV 254 nm. The resulting mixture was concentrated under vacuum. This yielded methyl 3-(4-methylpiperazin-1-yl)-2-(pyridin-3-yl)propionate I-18c (884 mg, 72.36%) as a yellow solid. LCMS: m / z (ESI), [M+H] + =264.30.

[0322] Step 3: At room temperature and under a nitrogen atmosphere, LiOH (68.2 mg, 2.85 mmol, 5 equivalents) was added in portions to a stirred mixture of piperazine I-18c (150 mg, 0.57 mmol, 1 equivalent) in THF (8 mL) and H2O (2 mL). The resulting mixture was stirred overnight at room temperature and under a nitrogen atmosphere. The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, water containing MeCN, 10% to 50% gradient, 10 min; detector, UV 254 nm. This yielded 3-(4-methylpiperazin-1-yl)-2-(pyridin-3-yl)propionic acid I-18 (120 mg, 84.5%) as a white solid. LCMS: m / z (ESI), [M+H] + =250.35.

[0323] Synthesis of intermediate I-19

[0324]

[0325] Step 1: At room temperature, add 200 mL of dioxane containing tert-butyl 3-bromobenzoate I-19a (8 g, 31.11 mmol, 1 equivalent), methyl cyanoacetate (4624.44 mg, 46.67 mmol, 1.5 equivalent), Pd(OAc)₂ (698.52 mg, 3.11 mmol, 0.1 equivalent), dppf (3437.21 mg, 6.22 mmol, 0.2 equivalent), and t-BuOK (10.47 g, 93.34 mmol, 3 equivalent) to a 500 mL round-bottom flask. Stir the resulting mixture at 70 °C under a nitrogen atmosphere for 6 hours. Acidify the mixture to pH 6 with saturated NH₄Cl (aqueous solution). Extract the mixture with ethyl acetate (3 x 50 mL). Dry the combined organic layers over anhydrous Na₂SO₄. After filtration, concentrate the filtrate under reduced pressure. The residue was purified by silica gel column chromatography using PE / EA (7:1, v / v) elution to obtain tert-butyl 3-(1-cyano-2-methoxy-2-oxoethyl)benzoate I-19b (3.7 g, 43.20%), which was a light yellow oil. 1 H NMR (CD3OD, 400MHz) δ1.63(9H,s),3.81(3H,s),7.56(1H,td),7.70(1H,ddd),8.02(1H,dt),8.05–8.09(1H,m).

[0326] Step 2: At room temperature, add 20 mL of methanol containing nitrile I-19b (3.7 g, 13.44 mmol, 1 equivalent) and CoCl2·6H2O (2.62 g, 20.16 mmol, 1.5 equivalent) and Boc2O (5.87 g, 26.88 mmol, 2 equivalent) to a 100 mL round-bottom flask. At 0 °C, add NaBH4 (1.53 g, 40.32 mmol, 3 equivalent) in portions over 30 minutes. Stir the resulting mixture at room temperature for another 15 hours. Quench the reaction with water at room temperature. Extract the mixture with DCM (3 x 40 mL). Dry the combined organic layers with anhydrous Na2SO4. After filtration, concentrate the filtrate under reduced pressure. The residue was purified by silica gel column chromatography using PE / EA (5:1, v / v) elution to obtain tert-butyl 3-{3-[(tert-butoxycarbonyl)amino]-1-methoxy-1-oxopropyl-2-yl}benzoate I-19c (2.07 g, 40.59%) as a white oil. LCMS: m / z (ESI), [M+H-Boc- t Bu] + =223.95.

[0327] Step 3: At room temperature, add 15 mL of water and 15 mL of methanol containing ester I-19c (2 g, 5.27 mmol, 1 equivalent) and LiOH (252.47 mg, 10.54 mmol, 2 equivalents) to a 100 mL round-bottom flask. Stir the resulting mixture for 2 hours at room temperature under a nitrogen atmosphere. Acidify the mixture to pH 5 with HCl (1 M). Purify the residue by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, methanol / water, 12% to 22% gradient, 25 min; detector, UV 254 nm. This yields 3-[(tert-butoxycarbonyl)amino]-2-[3-(tert-butoxycarbonyl)phenyl]propionic acid I-19 (650 mg, 33.75%) as a grayish-white solid. LCMS: m / z (ESI), [M+H] + =366.00.

[0328] Synthesis of intermediate I-20

[0329]

[0330] Step 1. At room temperature, add 20 mL of THF containing methyl 2-bromo-2-phenylacetate I-20a (1 g, 4.37 mmol, 1 equivalent), N-[2-(piperazin-1-yl)ethyl]carbamate tert-butyl ester (1.20 g, 5.24 mmol, 1.2 equivalent), and triethylamine (1.33 g, 13.10 mmol, 3 equivalent) to a 40 mL vial. Stir the resulting mixture at 70 °C under a nitrogen atmosphere for 15 hours. Quench the reaction by adding water (60 mL) at room temperature. Extract the resulting mixture with EA (3 x 50 mL). Wash the combined organic layers with saturated brine (1 x 100 mL) and dry over anhydrous Na₂SO₄. Filter and concentrate the filtrate under reduced pressure. Dissolve the residue in DCM (4 mL). The residue was purified by preparative TLC with PE / EA (1:1, v / v) to obtain methyl 2-(4-{2-[(tert-butoxycarbonyl)amino]ethyl}piperazin-1-yl)-2-phenylacetate I-20b (1.54 g, 93.45%), a pale yellow oil. LCMS: m / z (ESI), [M+H] + =378.10.

[0331] Step 2. At room temperature, add 5 mL of THF and 5 mL of water containing piperazine I-20b (600 mg, 1.59 mmol, 1 equivalent) and LiOH (76.14 mg, 3.18 mmol, 2 equivalents) to a 40 mL vial. Stir the resulting mixture for 15 hours at room temperature under a nitrogen atmosphere. Acidify the mixture to pH 5 with HCl (aqueous solution). Purify the residue by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, methanol / water, 5% to 10% gradient, 30 min; detector, UV 220 nm. Concentrate the resulting mixture under reduced pressure. This yields (4-{2-[(tert-butoxycarbonyl)amino]ethyl}piperazin-1-yl)(phenyl)acetic acid I-20 (469 mg, 81.18%) as a grayish-white solid. LCMS: m / z (ESI), [M+H] + =364.10. 1 H NMR (CD3OD, 400MHz) δ1.44(8H,s),2.63(2H,s),2.91(5H,d),3.22(2H,t),4.40(1H,s),7.18–7.49(3H,m),7.57(2H,dd).

[0332] Synthesis of intermediate I-21

[0333]

[0334] Step 1: At room temperature, a mixture of 3-(cyanomethyl)benzoic acid I-21a (5 g, 31.03 mmol, 1 equivalent) and CDI (5.53 g, 34.13 mmol, 1.1 equivalent) in THF (26 mL) was stirred for 3 hours. Under a nitrogen atmosphere at 0 °C, 20 mL of H₂O containing NaBH₄ (3.52 g, 93.08 mmol, 3 equivalent) was added dropwise to the mixture. The resulting mixture was stirred overnight under a nitrogen atmosphere at room temperature. The reaction was monitored by TLC at PE / EA (6:1, v / v). The reaction was quenched with water at room temperature. The resulting mixture was extracted with DCM (3 x 200 mL). The combined organic layers were dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using PE / EA (8:1, v / v) elution to give 2-[3-(hydroxymethyl)phenyl]acetonitrile (2.3 g, 50.37%) as a pale yellow liquid. 1 H NMR (CD3OD, 400MHz) δ3.91(2H,s),4.64(2H,s),7.00–7.07(1H,m),7.17(1H,t),7.39(1H,q),7.99(1H,s).

[0335] Step 2: At 0°C under a nitrogen atmosphere, TBSCl (2.83 g, 18.75 mmol, 1.2 equivalent) was added in portions to a stirred mixture of 2-[3-(hydroxymethyl)phenyl]acetonitrile (2.3 g, 15.63 mmol, 1 equivalent) and triethylamine (4.74 g, 46.88 mmol, 3 equivalent), and DMAP (0.02 g, 0.16 mmol, 0.01 equivalent) in a 10 mL DCM solution. The resulting mixture was stirred overnight at room temperature under a nitrogen atmosphere. The reaction was monitored by TLC at PE / EA (6:1, v / v). The reaction was quenched with water at room temperature. The resulting mixture was extracted with CH2Cl2 (3 x 100 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC with PE / EA (6:1, v / v) to obtain 2-(3-{[(tert-butyldimethylsilyl)oxy]methyl}phenyl)acetonitrile I-21b (2.1 g, 51.40%) as a pale yellow liquid. 1 H NMR (CD3OD, 400MHz) δ0.13(6H,s),0.97(9H,s),3.91(2H,s),4.78(2H,s),7.25(1H,d),7.30(1H,d),7.36(1H,d),7.38(1H,d).

[0336] Step 3. At 0°C under a nitrogen atmosphere, sodium hydride (289.14 mg, 12.05 mmol, 1.5 equivalents) was added in portions to a stirred mixture of nitrile I-21b (2.1 g, 8.03 mmol, 1 equivalent) in THF (25 mL). The resulting mixture was stirred for 1 hour at room temperature under a nitrogen atmosphere. Next, dimethyl carbonate (2.89 g, 32.13 mmol, 4 equivalents) was added in portions at 0°C under a nitrogen atmosphere. The final reaction mixture was stirred overnight at room temperature. The reaction was monitored by TLC at PE / EA (6:1, v / v). The reaction was quenched with water at room temperature. The resulting mixture was extracted with ethyl acetate (3 x 150 mL). The combined organic layers were dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC with PE / EA (6:1, v / v) to obtain methyl 2-(3-{[(tert-butyldimethylsilyl)oxy]methyl}phenyl)-2-cyanoacetate (1.9 g, 74.04%) as a pale yellow liquid. 1 H NMR (CD3OD, 400MHz) δ0.13(6H,s),0.97(9H,s),3.78(3H,s),4.80(2H,q),7.32–7.51(4H,m).

[0337] Step 4. At 0°C and under a nitrogen atmosphere, add NaBH4 (1.80 g, 47.58 mmol, 8 equivalents) in portions to a stirred mixture of methyl 2-(3-{[(tert-butyldimethylsilyl)oxy]methyl}phenyl)-2-cyanoacetate (1.9 g, 5.95 mmol, 1 equivalent) and (Boc)₂O (5.19 g, 23.79 mmol, 4 equivalents) and CoCl₂·6H₂O (8.9 g, 8.96 mmol, 1.5 equivalents) in methanol (50 mL). Stir the resulting mixture overnight at room temperature under a nitrogen atmosphere. Filter the mixture. Concentrate the filtrate under reduced pressure. The residue was purified by preparative TLC using PE / EA (5:1, v / v) to give methyl 3-[(tert-butoxycarbonyl)amino]-2-(3-{[(tert-butyldimethylsilyl)oxy]methyl}phenyl)propionate (1.2 g, 47.63%) as a pale yellow liquid. LCMS: m / z (ESI), [M+H] + =324.20.

[0338] Step 5. At 60°C under a nitrogen atmosphere, a solution containing methyl 3-[(tert-butoxycarbonyl)amino]-2-(3-{[(tert-butyldimethylsilyl)oxy]methyl}phenyl)propionate (1.2 g, 2.83 mmol, 1 equivalent) in methanol (1 mL) and H2O containing LiOH·H2O (237.72 mg, 5.67 mmol, 2 equivalents) (1 mL) was stirred overnight. The mixture was acidified to pH 5 with HCl (1 M). The resulting mixture was concentrated under vacuum. The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, methanol / water, 0% to 5% gradient, 30 min; detector, UV 220 nm. This yielded 3-[(tert-butoxycarbonyl)amino]-2-[3-(hydroxymethyl)phenyl]propionate I-21 (530 mg, 63.35%) as a grayish-white solid. LCMS:m / z(ESI),[M- t Bu] + =240.06.

[0339] Synthesis of intermediate I-22

[0340]

[0341] Step 1: Sodium hydride (2.55 g, 106.09 mmol, 1.2 equivalents) was added in portions to a solution of ethyl cyanoacetate (10 g, 88.41 mmol, 1.00 equivalents) in DMF (50 mL), and the mixture was stirred for 1 hour at 0 °C under a nitrogen atmosphere. Subsequently, dioxane (50 mL) containing 4-fluoronitrobenzene I-22a (8.73 g, 61.87 mmol, 0.70 equivalents) was added dropwise at 0 °C. The mixture was stirred for 12 hours at 80 °C. The mixture was concentrated under reduced pressure. The residue was dissolved in DCM (300 mL). The mixture was washed with 2 x 250 mL HCl (1 M), 2 x 250 mL water, and 1 x 250 mL saturated brine. The mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using PE / EA (8:1, v / v) elution to give ethyl 2-cyano-2-(4-nitrophenyl)acetate I-22b (11.8 g, 55.37%) as a red oil. LCMS: m / z (ESI), [M+H] + =235.10. 1 H NMR (400MHz, DMSO-d6) δ1.20(3H,t),4.15(2H,m),5.97(1H,s),7.74(2H,d),8.29(2H,d).

[0342] Step 2: At 0°C and under a nitrogen atmosphere, a solution of nitrile I-22b (1 g, 4.54 mmol, 1 equivalent) in methanol was treated with Boc₂O (3.96 g, 18.17 mmol, 4 equivalents) and CoCl₂·6H₂O (2.81 g, 11.81 mmol, 2.6 equivalents) for 5 minutes. Subsequently, NaBH₄ (1.37 g, 36.34 mmol, 8 equivalents) was added in portions at 0°C. The resulting mixture was stirred for 2 hours at room temperature. The reaction was quenched with water at room temperature. The resulting mixture was filtered, and the filter cake was washed with methanol (3 x 20 mL). The aqueous layer was extracted with CH₂Cl₂ (3 x 100 mL). The resulting mixture was concentrated under reduced pressure. The residue was purified by preparative TLC using CH₂Cl₂ / methanol (2:1, v / v) to give ethyl 3-((tert-butoxycarbonyl)amino)-2-(4-((tert-butoxycarbonyl)amino)phenyl)propionate I-22c (500 mg, 27.91%) as a yellow oil. LCMS: m / z (ESI), [M+H] + =409.25.

[0343] Step 3: At room temperature, add 5 mL of THF and 5 mL of H2O containing ester I-22c (500 mg, 1.27 mmol, 1 equivalent) and LiOH·H2O (106.37 mg, 2.54 mmol, 2 equivalents) to a 100 mL round-bottom flask. Stir the resulting mixture for 15 hours at room temperature under air atmosphere. Acidify the mixture to pH 5 with HCl (1 M). Concentrate the resulting mixture under reduced pressure. Dry the resulting oil by lyophilization. This yields 3-[(tert-butoxycarbonyl)amino]-2-{4-[(tert-butoxycarbonyl)amino]phenyl}propionic acid I-22 (420 mg, 68.74%) as a yellow solid. LCMS: m / z (ESI), [MH] - =379.15.

[0344] Synthesis of intermediate I-23

[0345]

[0346] Step 1: At room temperature and under a nitrogen atmosphere, a mixture of 2-(2-methoxyphenyl)acetonitrile I-23a (3 g, 20.38 mmol, 1 equivalent) and sodium hydride (391.04 mg, 16.31 mmol, 1.2 equivalent) in THF (30 mL) was stirred for 2 hours. At room temperature and under a nitrogen atmosphere, dimethyl carbonate (4.60 g, 50.96 mmol, 2.5 equivalent) was added dropwise to the mixture. At room temperature and under a nitrogen atmosphere, the resulting mixture was stirred for 2 hours. The mixture was quenched with water (40 mL). The aqueous layer was extracted with CH2Cl2 (3 x 20 mL) and dried over anhydrous Na2SO4. The resulting mixture was concentrated under reduced pressure. The crude product was purified rapidly by reverse-phase chromatography to give methyl 2-cyano-2-(2-methoxyphenyl)acetate (900 mg, 43.03%) as a yellow solid. LCMS:m / z(ESI),[M+H] + =206.21. 1 H NMR (400MHz, DMSO-d6) δ3.72(3H,s),3.83(3H,d),7.01(1H,t),7.12(1H,d),7.34(1H,d),7.42(1H,d).

[0347] Step 2: At room temperature and under a nitrogen atmosphere, CoCl2 (3.01 g, 23.15 mmol, 2.5 equivalents) and NaBH4 (2.80 g, 74.07 mmol, 8 equivalents) were added in portions to a stirred mixture of methyl 2-cyano-2-(2-methoxyphenyl)acetate (1.9 g, 9.26 mmol, 1 equivalent) and Boc2O (4.04 g, 18.52 mmol, 2 equivalents) in methanol. The resulting mixture was stirred for 2 hours at room temperature and under a nitrogen atmosphere. The mixture was quenched with water (30 mL). The aqueous layer was extracted with CH2Cl2 (3 x 20 mL) and dried over anhydrous Na2SO4. The resulting mixture was concentrated under reduced pressure. The crude product was purified by reverse-phase rapid purification to give methyl 3-[(tert-butoxycarbonyl)amino]-2-(2-methoxyphenyl)propionate I-23b (1.3 g, 45.39%) as a yellow solid. LCMS:m / z(ESI),[M+H] + =310.10. 1 H NMR(400MHz,DMSO-d6)δ1.34(9H,s),3.20(1H,d),3.50(1H,td),3.56(3H,s),3.76(3 H,s),4.11(1H,t),6.79(1H,t),6.91(1H,t),6.99(1H,d),7.15(1H,d),7.26(1H,d).

[0348] Step 3: At room temperature and under a nitrogen atmosphere, a mixture of methyl 3-[(tert-butoxycarbonyl)amino]-2-(2-methoxyphenyl)propionate (1.3 g, 4.20 mmol, 1 equivalent) and LiOH (0.40 g, 16.81 mmol, 4 equivalents) in methanol (4 mL) and water (1 mL) was stirred for 2 hours. The resulting mixture was concentrated under reduced pressure. The crude product was rapidly purified by reverse-phase chromatography to give 3-[(tert-butoxycarbonyl)amino]-2-(2-methoxyphenyl)propionate I-23 (800 mg, 64.46%) as a yellow solid. 1 H NMR(400MHz, DMSO-d6)1.33(9H,s),3.08–3.24(2H,m),3.75(4H,s),6.61(1H,s),6.78–6.87(1H,m),6.84–6.97(1H,m),7.13(1H,td),7.22(1H,dd).

[0349] Synthesis of intermediate I-24

[0350]

[0351] Step 1: A solution of 4-fluorophenylacetonitrile I-24a (3 g, 22.20 mmol, 1 equivalent) in THF was treated with sodium hydride (1.1 g, 45.84 mmol, 2.06 equivalent) for 1 hour at 0 °C under a nitrogen atmosphere. Subsequently, dimethyl carbonate (4.40 g, 48.84 mmol, 2.2 equivalent) was added in portions at room temperature. The resulting mixture was stirred for 12 hours at room temperature. The reaction was quenched with water at room temperature. The resulting mixture was extracted with ethyl acetate (3 x 30 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC with PE / EA (3:1, v / v) to give methyl 2-cyano-2-(4-fluorophenyl)acetate (1.1 g, 15.01%) as a yellow oil. 1 H NMR (DMSO-d6, 400MHz) δ3.74(3H,s),5.73(1H,s),7.25–7.35(2H,m),7.46–7.55(2H,m).

[0352] Step 2: At 0°C and under a nitrogen atmosphere, a mixture of methyl 2-cyano-2-(4-fluorophenyl)acetate (1 g, 5.18 mmol, 1 equivalent), Boc₂O (2.26 g, 10.36 mmol, 2.00 equivalent), and CoCl₂·6H₂O (2.46 g, 10.34 mmol, 2.00 equivalent) in methanol was stirred for 5 minutes. NaBH₄ (1.57 g, 41.42 mmol, 8 equivalent) was added to the mixture at 0°C to 10°C. The resulting mixture was filtered, and the filter cake was washed with CH₂Cl₂ (3 x 30 mL). The filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC with PE / EA (5:1, v / v) to give methyl 3-[(tert-butoxycarbonyl)amino]-2-(4-fluorophenyl)propionate I-24b (400 mg, 25.68%) as a yellow oil. LCMS:m / z(ESI),[M+H] + =298.25.

[0353] Step 3. At room temperature, methyl 3-[(tert-butoxycarbonyl)amino]-2-(4-fluorophenyl)propionate I-24b (380 mg, 1.28 mmol, 1 equivalent), LiOH (61.22 mg, 2.56 mmol, 2.00 equivalent), THF (2 mL), and water (1 mL) were added to a 50 mL round-bottom flask. The resulting mixture was stirred at 60 °C for 8 hours. The mixture was acidified to pH 4–6 with concentrated HCl. The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, methanol / water, 10% to 50% gradient, 10 min; detector, UV 254 nm. This yielded 3-[(tert-butoxycarbonyl)amino]-2-(4-fluorophenyl)propionate I-24 (260 mg, 63.98%) as a yellow solid. LCMS: m / z (ESI), [M-56] + =228.05.

[0354] Synthesis of intermediate I-25

[0355]

[0356] Step 1: To a solution of (1H-benzo[d][1,2,3]triazol-1-yl)methanol I-25a (25 g, 167.62 mmol) in toluene (400 mL), tert-butyl carbamate (19.64 g, 167.62 mmol) and 4-methylbenzenesulfonic acid hydrate (64 mg, 336.46 μmol, 51.61 μL) were added, and the solution was refluxed at 120 °C using a Dean-Stark separator for 24 hours. Half of the toluene was evaporated under reduced pressure, the solution was cooled to 0 °C, and the product was recrystallized to give N-(benzotriazol-1-ylmethyl)carbamate tert-butyl I-25b (24 g, 58% yield) as a white solid. LCMS:t R =1.55 min, 3 min chromatography (3 min - 5 - 95% MeCN / water (6 mmol / L NH4HCO3), Waters Acquity UPLC BEH C18 1.7 μm, 2.1*50 mm, 40 °C), MS (ESI) m / z = 249.1 [M+H] + . 1 H NMR: (400MHz, DMSO-d6) δ8.42(t,J=6.4Hz,1H),8.04(d,J=8.4Hz,1H),7.96(d,J=8.4 Hz,1H),7.57(t,J=7.6Hz,1H),7.44–7.38(m,1H),5.88(d,J=6.7Hz,2H),1.37(s,9H).

[0357] Step 2: Slowly add 2.5M n-butyllithium (3.32g, 51.87mmol, 20.75mL) to 100mL THF containing (R)-4-benzyloxazolidin-2-one (9.1g, 51.35mmol) at -78°C along the side of the flask, and stir the reaction mixture for 20 minutes at -78°C. Next, add the solution of (R)-4-benzyloxazolidin-2-one containing lithium anions in THF through a tube to a pre-cooled (-78°C) solution of 2-(3-methoxyphenyl)acetyl chloride I-25c (10.05g, 54.44mmol) dissolved in 100mL THF. Stir the reaction mixture for 1 hour at -78°C, then pour it into NH4Cl (saturated) / HCl (1M) (3:1) and extract with ethyl acetate. Dry the combined extracts over anhydrous sodium sulfate, filter, and evaporate under reduced pressure. The residue was purified by rapid silica gel column chromatography with elution using a gradient of ethyl acetate / petroleum ether from 0% to 40%. The purified fraction was lyophilized to give (4R)-4-benzyl-3-[2-(3-methoxyphenyl)acetyl]oxazolidin-2-one I-25d (12.19 g, 73% yield) as a white solid. LCMS:t R=1.82 min, 3 min chromatography (3 min - 5 - 95% MeCN / water (6 mmol / L NH4HCO3), WatersAcquity UPLC BEH C18 1.7 μm, 2.1*50 mm, 40 °C), MS (ESI) m / z = 326.2 [M+H] + . 1 H NMR: (400MHz, DMSO-d6) δ7.32–7.20(m,4H),7.19–7.08(m,2H),6.86(d,J=7.1Hz,3H),4.68(ddd,J=10.8,7.6,2.9Hz,1H),4.34(t,J=8. 5Hz, 1H), 4.25 (d, J = 15.9Hz, 1H), 4.20 (dd, J = 8.9, 2.8Hz, 1H), 4.10 (d, J = 15.9Hz, 1H), 3.75 (s, 3H), 2.95 (ddd, J = 20.9, 13.5, 5.3Hz, 2H).

[0358] Step 3: Add (4R)-4-benzyl-3-[2-(3-methoxyphenyl)acetyl]oxazolidin-2-one I-25d (12.19 g, 37.47 mmol) in THF (60 mL) cooled to -70 °C to a solution of 1 M LiHMDS (10.68 g, 44.59 mmol) in THF (44.59 mL) via a tube, and stir this mixture at -70 °C for 40 minutes. Then, add N-(benzotriazol-1-ylmethyl)carbamate tert-butyl ester I-25b (11.16 g, 44.96 mmol) in THF (60 mL), also cooled to -70 °C, via a tube. Warm this solution to -25 to -30 °C and stir for 2 hours. The mixture was poured into ethyl acetate and NH4Cl (saturated) / HCl (1M) (3:1), and the aqueous phase was further extracted with ethyl acetate at least once. The combined organic matter was dried over anhydrous sodium sulfate, filtered, and evaporated under reduced pressure. The residue was purified by passing it through a silica gel column and eluting with a gradient of ethyl acetate / DCM from 0 to 4%. The purified fraction was lyophilized to give N-[(2S)-3-[(4R)-4-benzyl-2-oxo-oxazolidine-3-yl]-2-(3-methoxyphenyl)-3-oxo-propyl] tert-butyl carbamate I-25e (15 g, 88% yield) as a soft white solid. LCMS:t R=1.95 min, 3 min chromatography (3 min - 5 - 95% MeCN / water (0.02% TFA), Waters Acquity UPLC BEH C18 1.7 μm, 2.1*50 mm, 40 °C), MS (ESI) m / z = 455.3 [M+H] + . 1 H NMR: (500MHz, DMSO-d6)δ7.37–7.17(m,6H),7.08(t,J=5.6Hz,1H),6.83(t,J =7.1Hz,2H),6.78(s,1H),5.16(dd,J=8.4,6.1Hz,1H),4.68(t,J=7.6Hz,1H), 4.25(t,J=8.5Hz,1H),4.17(dd,J=12.4,10.3Hz,1H),3.72(s,3H),3.67–3.57 (m,1H),3.29–3.17(m,1H),3.02(ddd,J=20.6,13.5,4.9Hz,2H),1.35(s,9H).

[0359] Step 4: To a solution of N-[(2S)-3-[(4R)-4-benzyl-2-oxo-oxazolidine-3-yl]-2-(3-methoxyphenyl)-3-oxo-propyl] tert-butyl carbamate I-25e (15 g, 33.00 mmol) cooled to 0 °C, add 30% hydrogen peroxide (16.36 g, 144.26 mmol, 14.87 mL, 30% purity) and lithium hydroxide (1.03 g, 42.90 mmol). Stir the solution at 0 °C for 0.75 h and add 400 mL of saturated sodium sulfate solution. Stir the mixture for another 15 min, then acidify to pH 5 with 1 M HCl (pH paper) and extract with ethyl acetate (200 mL x 3). The combined organic matter is dried over anhydrous sodium sulfate, filtered, and evaporated under reduced pressure. The residue was purified by rapid silica gel column chromatography and eluted with a gradient of 0 to 40% ethyl acetate / petroleum ether containing 1% acetic acid. The purified fraction was lyophilized to give (2S)-3-(tert-butoxycarbonylamino)-2-(3-methoxyphenyl)propionic acid I-25 (8.7 g, 89% yield) as a white solid. 1 H NMR: (500MHz, DMSO-d6)δ12.33(s,1H),7.24(t,J=7.9Hz,1H),3.79–3.63(m,4H),3.74-3 .71(m,4H)3.42(ddd,J=13.3,7.8,5.4Hz,1H),3.21(dt,J=13.3,6.7Hz,1H),1.33(s,9H). SFCt R=4.638 minutes, ee value = 98.7%.

[0360] Synthesis of intermediate I-26

[0361]

[0362] Step 1: At 0°C, sodium hydride (2.45 g, 102.02 mmol, 2 equivalents) was added to a stirred mixture of 4-bromo-phenylacetonitrile I-26a (10 g, 51.01 mmol, 1 equivalent) in THF (200 mL). The resulting mixture was stirred at 0°C under air atmosphere for 2 hours. Then, dimethyl carbonate (18.38 g, 204.03 mmol, 4 equivalents) was added to the mixture. After the reaction, the mixture was quenched with ice water (200 mL). The mixture was extracted with ethyl acetate (3 x 100 mL). The combined organic layers were dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using PE / EA (10:1, v / v) elution to give methyl 2-(4-bromophenyl)-2-cyanoacetate I-26b (6.1 g, 47.07%) as a yellow oil. 1 H NMR (DMSO-d6, 400MHz) δ3.74(3H,s),5.74(1H,s),7.41(2H,m),7.68(2H,m).

[0363] Step 2: CoCl₂·6H₂O (1102.40 mg, 39.36 mmol, 2 equivalents) and (Boc)₂O (8589.67 mg, 39.36 mmol, 2 equivalents) were added to a stirred mixture of nitrile I-26b (5.0 g, 19.68 mmol, 1 equivalent) in methanol (35 mL). NaBH₄ (4466.65 mg, 118.07 mmol, 6 equivalents) was added at 0 °C. The resulting mixture was stirred at 25 °C under air for 4 hours. After the reaction, the mixture was quenched with 100 mL of ice water. The mixture was extracted with ethyl acetate (3 x 70 mL). The combined organic layers were dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using PE / EA (5:1, v / v) elution to obtain methyl 2-(4-bromophenyl)-3-[(tert-butoxycarbonyl)amino]propionate I-26c (4.0 g, 56.74%) as a yellow oil. 1H NMR (DMSO-d6, 400MHz) δ1.34(9H,d),3.26(1H,m),3.48(1H,m),3.60(3H,d),3.85(1H,t),6.94(1H,t),7.24(2H,m),7.34(1H,m),7.53(1H,m).

[0364] Step 3: Add 4 mL of water containing LiOH·H₂O (1874.09 mg, 44.66 mmol, 4 equivalents) to a stirred mixture of ester I-26c (4.0 g, 11.17 mmol, 1 equivalent) in methanol (16 mL). Stir the resulting mixture at 60 °C under a nitrogen atmosphere for 8 hours. After cooling to 25 °C, concentrate the resulting mixture under reduced pressure. Dilute the reaction mixture with water (20 mL) and adjust the pH to 2 with HCl (1 M, 30 mL). Purify the residue by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, phase A: water; phase B: ACN, 0% to 50% gradient, 40 min; detector, UV 254 nm, 30% hold for 20 min. After evaporation of the solvent, 2-(4-bromophenyl)-3-[(tert-butoxycarbonyl)amino]propionic acid I-26 (3.2 g, 83.26%) is given as a yellow solid. LCMS:m / z(ESI),[M- t Bu] + =289.95.

[0365] Synthesis of intermediate I-27

[0366]

[0367] Step 1: At room temperature and under an air atmosphere, Dppf (480.33 mg, 0.87 mmol, 0.2 equivalent) was added to a stirred solution of 1-bromo-4-(2-methoxyethoxy)benzene I-27a (1 g, 4.35 mmol, 1 equivalent) and methyl cyanoacetate (198.18 mg, 2.00 mmol, 0.46 equivalent), Pd(OAc)2 (97.61 mg, 0.44 mmol, 0.1 equivalent), and t-BuOK (975.77 mg, 8.70 mmol, 2 equivalent) in dioxane (15 mL). The mixture was stirred overnight at 70 °C under a nitrogen atmosphere. The resulting mixture was extracted with ethyl acetate (3 x 20 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC with PE / EA (1:1, v / v) to obtain methyl 2-cyano-2-[4-(2-methoxyethoxy)phenyl]acetate (200 mg, 9.26%) as a yellow oil. 1H NMR (CDCl3, 400MHz) δ3.48(3H,s),3.76–3.80(2H,m),3.82(3H,s),4.12–4.17(2H,m),6.98(2H,d),7.39(2H,d).

[0368] Step 2: At 0°C and under air, NaBH4 (364.24 mg, 9.63 mmol, 8 equivalents) was added in portions to a stirred mixture of methyl 2-cyano-2-[4-(2-methoxyethoxy)phenyl]acetate (300 mg, 1.20 mmol, 1 equivalent), CoCl2·6H2O (572.69 mg, 2.41 mmol, 2 equivalents), and (Boc)2O (525.34 mg, 2.41 mmol, 2.00 equivalents) in methanol (10 mL). The methanol mixture containing the mixture was stirred overnight at 0°C and under air. The reaction was quenched with water (10 mL) at room temperature. The resulting mixture was extracted with ethyl acetate (3 x 15 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC with CH₂Cl₂ / methanol (15:1, v / v) to give methyl 3-[(tert-butoxycarbonyl)amino]-2-[4-(2-methoxyethoxy)phenyl]propionate I-27b (100 mg, 23.51%) as a white solid. LCMS: m / z (ESI), [M+H-boc] + =254.

[0369] Step 3: At 0°C under a nitrogen atmosphere, LiOH (40.66 mg, 1.70 mmol, 3 equivalents) was added in portions to a stirred mixture of ester I-27b (200 mg, 0.57 mmol, 1 equivalent) in THF (40 mL) and H₂O (10 mL). The resulting mixture was stirred overnight at 60°C under a nitrogen atmosphere. The mixture was cooled to room temperature. The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water (0.1% FA), 10% to 50% gradient, 10 min; detector, UV 254 nm. This yielded 3-[(tert-butoxycarbonyl)amino]-2-[4-(2-methoxyethoxy)phenyl]propionic acid I-27 (90 mg, 46.86%) as a white solid. LCMS: m / z (ESI), [M+H-boc] + =240.

[0370] Synthesis of intermediate I-28

[0371]

[0372] Step 1: At 0°C, an oil containing 60% sodium hydride (1.47 g, 61.15 mmol, 1 equivalent) was added to a solution of 2-(4-methoxyphenyl)acetonitrile I-28a (6 g, 40.77 mmol, 1 equivalent) in THF (100 mL). The mixture was stirred for 80 minutes. At 0°C, dimethyl carbonate (29.38 g, 326.14 mmol, 8 equivalent) was added, and the mixture was warmed to RT and stirred for 4 hours. The reaction mixture was quenched with water (100 mL) and extracted with DCM (3 x 100 mL). The residue was purified by silica gel column chromatography using PE / EA (8:1, v / v) elution to give methyl 2-cyano-2-(4-methoxyphenyl)acetate (5.96 g, 70.03%) as a yellow oil.

[0373] Step 2: NaBH4 (40.56 g, 107.21 mmol, 4 equivalents) was added fractionally to a mixture of methyl 2-cyano-2-(4-methoxyphenyl)acetate (5.50 g, 26.80 mmol, 1 equivalent), (Boc)₂O (11.7 g, 53.60 mmol, 2 equivalents), and CoCl₂·6H₂O (8.29 mg, 34.84 mmol, 1.3 equivalents) in methanol (10 mL). The mixture was stirred overnight at room temperature under a nitrogen atmosphere. The reaction was quenched with water (50 mL) at 0 °C. The resulting mixture was extracted with DCM (3 x 40 mL). The combined organic layers were washed with saturated brine (1 x 100 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using PE / EA (5:1, v / v) elution to obtain methyl 3-[(tert-butoxycarbonyl)amino]-2-(4-methoxyphenyl)propionate I-28b (3.50 mg, 38.16%) as a colorless oil. LCMS: m / z (ESI), [M+H] + =310.15.

[0374] Step 3: At room temperature, ester I-28b (2 g, 6.47 mmol, 1 equivalent) and LiOH·H2O (0.54 g, 12.93 mmol, 2 equivalents) and H2O (4 mL) and methanol (8 mL) were added to a 50 mL round-bottom flask. The resulting mixture was stirred for 2 hours at room temperature under an air atmosphere. The residue was acidified to pH 5 with HCl (1 M). The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, methanol / water, 0% to 15% gradient, 30 min; detector, UV 220 nm. The resulting liquid was dried by lyophilization. This yielded 3-[(tert-butoxycarbonyl)amino]-2-(4-methoxyphenyl)propionic acid I-28 (1.38 g, 70.83%) as a white solid. LCMS: m / z (ESI), [M+H- t Bu] + =240.25.

[0375] Synthesis of intermediate I-29

[0376]

[0377] Step 1: At 0°C and under air, sodium hydride (181.57 mg, 7.57 mmol, 2 equivalents) was added in portions to a stirred solution of 2-(6-methylpyridin-2-yl)acetonitrile I-29a (500 mg, 3.78 mmol, 1 equivalent) in THF (20 mL). The resulting mixture was stirred for 1 hour at room temperature under air. Dimethyl carbonate (436.34 mg, 4.54 mmol, 1.2 equivalents) was added to the mixture at room temperature. The mixture was stirred for another 2 hours at room temperature. The reaction was quenched by adding water / ice (20 mL) at room temperature. The mixture was extracted with CH2Cl2 (3 x 20 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC using CH₂Cl₂ / methanol (20:1, v / v) to give methyl 2-cyano-2-(6-methylpyridin-2-yl)acetate (330 mg, 45.86%) as an orange solid. LCMS: m / z (ESI), [M+H] + =191.20. 1 H NMR (CDCl3, 400MHz) δ2.49(3H,s),3.82(3H,s),6.49(1H,d),7.18(1H,d),7.54(1H,dd).

[0378] Step 2: At room temperature and under air atmosphere, CoCl₂·6H₂O (6.75 g, 28.39 mmol, 3 equivalents) and (Boc)₂O (6.20 g, 28.39 mmol, 3 equivalents) were added dropwise to a stirred solution of methyl 2-cyano-2-(6-methylpyridin-2-yl)acetate (1.8 g, 9.46 mmol, 1 equivalent) in methanol (20 mL). NaBH₄ (2.86 g, 75.71 mmol, 8 equivalents) was added dropwise to the mixture at 0 °C. The resulting mixture was stirred overnight at room temperature. The reaction was quenched by adding water (20 mL) at room temperature. The mixture was extracted with CH₂Cl₂ (3 x 20 mL). The resulting mixture was concentrated under reduced pressure. The residue was purified by preparative TLC with CH2Cl2 / methanol (50:1, v / v) to give methyl 3-[(tert-butoxycarbonyl)amino]-2-(6-methylpyridin-2-yl)propionate I-29b (1 g, 35.90%) as a yellow oil. 1 H NMR (DMSO-d6, 400MHz) δ1.34(9H,s),2.43(3H,s),3.31–3.44(1H,m),3.50–3.60(4H,m),3.99(1H,t),6.87(1H,t),7.13(2H,dd),7.65(1H,t).

[0379] Step 3: At room temperature, LiOH (0.16 g, 6.69 mmol, 1.97 equivalent) was added to a stirred solution of ester I-29b (1 g, 3.40 mmol, 1 equivalent) in methanol (20 mL) and H2O (10 mL). The resulting mixture was stirred at 60 °C under air atmosphere for 2 hours. The resulting mixture was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water, 10% to 20% gradient, 10 min; detector, UV 254 nm, to give 3-[(tert-butoxycarbonyl)amino]-2-(6-methylpyridin-2-yl)propionic acid I-29 (865 mg, 90.83%) as a yellow solid. 1 H NMR (DMSO-d6, 400MHz) δ1.34(9H,s),2.41(3H,s),3.34–3.38(2H,m),3.45(1H,t),6.51(1H,t),6.99(1H,d),7.09(1H,d),7.51(1H,t).

[0380] Synthesis of intermediate I-30

[0381]

[0382] Step 1: At room temperature and under air, K₂CO₃ (11.00 g, 79.56 mmol, 3 equivalents) was added to a stirred mixture of ethyl cyanoacetate (3 g, 26.52 mmol, 1 equivalent) and bromocyclopentane I-30a (4.74 g, 31.83 mmol, 1.2 equivalents) in DMSO (50 mL). The resulting mixture was stirred at 60 °C under air for 2 hours. After cooling to room temperature, the reaction was quenched by adding water (20 mL) at room temperature. The resulting mixture was extracted with ethyl acetate (3 x 20 mL). The combined organic layers were washed with brine (3 x 20 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water, 40% to 50% gradient, 10 min; detector, UV 254 nm, to obtain ethyl 2-cyano-2-cyclopentylacetate (1.1 g, 22.89%) as a yellow oil. 1 H NMR(DMSO-d6,400MHz)δ1.23(3H,t),1.28-1.41(2H,m),1.46-1.60(2H,m),1.60 -1.71(2H,m),1.71-1.90(2H,m),2.40(1H,ttd),4.19-4.26(2H,m).

[0383] Step 2: At room temperature and under air, (Boc)₂O (3.25 g, 14.90 mmol, 3 equivalents) was added dropwise to a stirred mixture of ethyl 2-cyano-2-cyclopentylacetate (900 mg, 4.97 mmol, 1 equivalent) and CoCl₂·6H₂O (3.54 g, 14.90 mmol, 3 equivalents) in methanol (20 mL). NaBH₄ (1.50 g, 39.73 mmol, 8 equivalents) was added in portions over 3 minutes at 0 °C. The resulting mixture was stirred overnight at room temperature and under air. The reaction was quenched by adding water / ice (20 mL) at 0 °C, and the mixture was concentrated under reduced pressure. The mixture was extracted with ethyl acetate (3 x 20 mL). The combined organic layers were washed with brine (3 x 20 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using PE / EA (1:1, v / v) elution to obtain ethyl 3-[(tert-butoxycarbonyl)amino]-2-cyclopentylpropionate I-30b (900 mg, 63.51%) as a yellow oil. 1H NMR(DMSO-d6,400MHz)δ1.18(3H,s),1.36(9H,s),1.50–1.67(6H,m),1.77–1.88(2H ,m),2.35(1H,td),3.02–3.13(2H,m),3.13–3.19(1H,m),4.05(2H,d),6.86(1H,t).

[0384] Step 3: At room temperature and under air atmosphere, LiOH (184.63 mg, 7.71 mmol, 2 equivalents) was added to a stirred solution of ester I-30b (1.1 g, 3.85 mmol, 1 equivalent) in methanol (10 mL) and H2O (5 mL). The resulting mixture was stirred at 60 °C under air atmosphere for 2 hours. After cooling to room temperature, the resulting mixture was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water, 20% to 30% gradient, 10 min; detector, UV 254 nm, to give 3-[(tert-butoxycarbonyl)amino]-2-cyclopentylpropionic acid I-30 (878 mg, 88.52%) as a white solid. 1 H NMR(DMSO-d6,400MHz) δ1.36(9H,s),1.57(8H,d),1.83(1H,s),3.00(2H,s),6.52(1H,s).

[0385] Synthesis of intermediate I-31

[0386]

[0387] Step 1: At room temperature, add 10 mL of methanol containing methyl 3-[(tert-butoxycarbonyl)amino]-2-(3-cyanophenyl)propionate I-3d (300 mg, 0.99 mmol, 1 equivalent), 2.5 mL of NH3 aqueous solution (64.20 mmol, 65.13 equivalent), and Raney Ni (10 mg, 0.12 mmol, 0.12 equivalent) to a 50 mL round-bottom flask. Stir the resulting mixture overnight at room temperature under a H2 atmosphere. Filter the mixture and wash the filter cake with methanol (2 x 20 mL). Concentrate the filtrate under reduced pressure to give methyl 2-[3-(aminomethyl)phenyl]-3-[(tert-butoxycarbonyl)amino]propionate I-31b (260 mg, 77.94%) as a green oil. LCMS: m / z (ESI), [M+H] + =309.15.

[0388] Step 2: At 25°C and under a N2 atmosphere, triethylamine (157.51 mg, 1.56 mmol, 1 equivalent) and MgSO4 (93.67 mg, 0.78 mmol, 1 equivalent) were added to a solution of amine I-31b (240 mg, 0.78 mmol, 20 equivalent) in DCM (100 mL). The mixture was stirred at 25°C for 30 minutes. Then, at 25°C, NaBH(OAc)3 (659.78 mg, 3.11 mmol, 4 equivalent) was added to the resulting mixture. The mixture was stirred at 25°C for 12 hours. The resulting mixture was filtered, and the filter cake was washed with methanol (3 x 100 mL). The filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC with PE / EA (3:1, v / v) to obtain methyl 3-[(tert-butoxycarbonyl)amino]-2-{3-[(dimethylamino)methyl]phenyl}propionate I-31c (115 mg, 37.24%) as a yellow oil. LCMS: m / z (ESI), [M+H] + =337.20.

[0389] Step 3: At 25°C and under a N2 atmosphere, LiOH·H2O (26.19 mg, 0.62 mmol, 2 equivalents) was added to a solution of ester I-31c (105 mg, 0.31 mmol, 1 equivalent) in THF (20 mL) and H2O (4 mL). The mixture was stirred at 25°C for 2 hours. The resulting mixture was concentrated under reduced pressure. This yielded 3-[(tert-butoxycarbonyl)amino]-2-{3-[(dimethylamino)methyl]phenyl}propionic acid I-31 (100 mg, 69.15%) as a white solid. LCMS: m / z (ESI), [M+H] + =323.20.

[0390] Synthesis of intermediate I-32

[0391]

[0392] Step 1: Dimethylphosphine oxide (614.37 mg, 7.87 mmol, 1 equivalent) of Pd(OAc)₂ (176.72 mg, 0.79 mmol, 0.2 equivalent), XantPhos (683.20 mg, 1.18 mmol, 0.3 equivalent), and K₃PO₄ (2088.54 mg, 9.84 mmol, 2.5 equivalent) were added to a stirred mixture of bromide I-26b (1.0 g, 3.94 mmol, 10 mL) and 1,4-dioxane (10 mL). The mixture was stirred at 80 °C under a nitrogen atmosphere for 2 hours. After cooling to 25 °C, the mixture was diluted with water (40 mL). The mixture was extracted with ethyl acetate (3 x 50 mL). The combined organic layers were washed with water (3 x 30 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. Then, it was diluted with DMF (5 mL). The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water, 10% to 50% gradient, 10 min; detector, UV 220 nm, 15% hold for 15 min. After solvent evaporation, methyl 2-cyano-2-[4-(dimethylphosphoryl)phenyl]acetate (220 mg, 22.25%) was obtained as a yellow solid. LCMS: m / z (ESI), [M+H) + =252.05.

[0393] Step 2: At 0°C and under a nitrogen atmosphere, NaBH4 (795.08 mg, 21.02 mmol, 8 equivalents) was added in portions to a stirred mixture of methyl 2-cyano-2-[4-(dimethylphosphoryl)phenyl]acetate (660 mg, 2.63 mmol, 1 equivalent), CoCl2·6H2O (1875.16 mg, 7.881 mmol, 3 equivalents), and (Boc)2O (1146.75 mg, 5.25 mmol, 2 equivalents) in MeOH (40 mL). The resulting mixture was stirred overnight. After the reaction, the reaction mixture was quenched by adding ice water (40 mL). The resulting mixture was extracted with DCM (3 x 40 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC with PE / EA (5:1, v / v) to give methyl 3-[(tert-butoxycarbonyl)amino]-2-[4-(dimethylphosphoryl)phenyl]propionate I-32b (330 mg, 35.35%) as a yellow oil. LCMS: m / z (ESI), [M+H- t Bu] + =300.10.

[0394] Step 3: At 25°C under a nitrogen atmosphere, water (1 mL) containing LiOH·H₂O (146.41 mg, 3.49 mmol, 4 equivalents) was added dropwise to a stirred mixture of ester I-32b (310 mg, 0.87 mmol, 1 equivalent) in methanol (4 mL). The resulting mixture was stirred overnight at 60°C under a nitrogen atmosphere. After cooling to 25°C, the resulting mixture was concentrated under reduced pressure and diluted with methanol (5 mL). The solution was acidified with HCl (1 M, 10 mL). DCM (10 mL) was added to the above solution. After extraction, the reaction mixture was evaporated under reduced pressure. The residue was diluted with DMF (2 mL) and purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, phase B MeCN, phase A water, 10% to 50% gradient, 10 min; detector, UV 254 nm, 16.5% hold for 20 min. After lyophilization, this yielded 3-[(tert-butoxycarbonyl)amino]-2-[4-(dimethylphosphoryl)phenyl]propionic acid I-32 (200 mg, 67.17%) as a white solid. 1 H NMR(DMSO-d6,400MHz)δ1.33(9H,s),1.60(6H,d),3.27(2H,m),7.37(2H,dd),7.60(2H,m).

[0395] Synthesis of intermediate I-33

[0396]

[0397] Step 1: At 0°C, sodium hydride (812.53 mg, 33.86 mmol, 2 equivalents) was added to a stirred solution of 2-(pyridin-2-yl)acetonitrile I-33a (2.0 g, 16.93 mmol, 1.00 equivalents) in THF (30 mL). The resulting mixture was stirred at 0°C under a nitrogen atmosphere for 30 minutes. Dimethyl carbonate (3049.90 mg, 33.86 mmol, 2 equivalents) was added dropwise to the mixture at 0°C. The resulting mixture was stirred again at 25°C for 3 hours. The reaction was quenched by adding water (80 mL) at 0°C. The resulting mixture was extracted with ethyl acetate (3 x 60 mL). The combined organic layers were washed with water (3 x 40 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using DCM / methanol (6%) elution to give methyl 2-cyano-2-(pyridin-2-yl)acetate (2.3 g, 77.12%) as a yellow solid. LCMS: m / z (ESI), [M+H] + =176.95. 1H NMR(DMSO-d6,400MHz)δ3.69(3H,s),6.88(1H,t),7.21(1H,d),7.84(1H,ddd),8.09(1H,t).

[0398] Step 2: At 0 °C, NaBH4 (2898.85 mg, 76.63 mmol, 6 equivalents) was added to a stirred mixture of methyl 2-cyano-2-(pyridin-2-yl)acetate (2.25 g, 12.77 mmol, 1 equivalent), CoCl2·6H2O (6077.14 mg, 25.54 mmol, 2 equivalents) and (Boc)2O (5574.69 mg, 25.54 mmol, 2 equivalents) in methanol (50 mL). The resulting mixture was stirred for 3 hours at 25 °C under air atmosphere. The reaction was quenched by adding water (80 mL) at 0 °C. After filtration, the mixture was extracted with DCM (60 mL x 3). The combined organic layers were washed with water (3 x 50 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC with PE / EA (3 / 1, v / v) to give methyl 3-[(tert-butoxycarbonyl)amino]-2-(pyridin-2-yl)propionate I-33b (600 mg, 16.76%) as a yellow solid. LCMS: m / z (ESI), [M+H] + =281.05. 1 H NMR (DMSO-d6, 400MHz) δ1.34(9H,s),3.33(1H,s),3.42(1H,dt),3.55(3H,dd),4.04(1H,q),6.87(1H,t),7.30(2H,m),7.77(1H,td),8.51(1H,m).

[0399] Step 3: At 25°C, add 1.0 mL of water containing LiOH (196.50 mg, 8.20 mmol, 4 equivalents) to a stirred mixture of ester I-33b (575 mg, 2.05 mmol, 1 equivalent) in methanol (4.0 mL). Stir the resulting mixture at 70°C under air atmosphere for 3 hours. After cooling to room temperature, concentrate the resulting mixture under vacuum and dilute with methanol (5 mL). Acidify the solution with HCl (1 M, 10 mL). Add DCM (10 mL) to the above solution. After extraction, evaporate the reaction mixture under reduced pressure. Dilute the residue with DMF (2 mL). Purify the residue by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, phase B MeCN, phase A water, 10% to 50% gradient, 10 min; detector, UV 254 nm, 10.8% hold for 20 min. After lyophilization, 3-[(tert-butoxycarbonyl)amino]-2-(pyridin-2-yl)propionic acid I-33 (387 mg, 70.85%) was obtained as a white solid. LCMS: m / z (ESI), [M+H] + =267.00. 1 H NMR (DMSO-d6, 400MHz) δ1.33(9H,s),3.39(1H,d),3.43(1H,m),3.54(1H,t),6.50(1H,t),7.15(1H,m),7.32(1H,dd),7.63(1H,td),8.40(1H,m).

[0400] Synthesis of intermediate I-34

[0401]

[0402] Step 1: At 0°C and under air, a solution of (4-bromo-2-methoxyphenyl)methanol I-34a (2 g, 9.21 mmol, 1 equivalent) in CH₂Cl₂ was treated with triethylamine (2.80 g, 27.64 mmol, 3 equivalents) and DMAP (11.26 mg, 0.092 mmol, 0.01 equivalents) for 5 minutes. Subsequently, TBSCl (1.67 g, 11.06 mmol, 1.2 equivalents) was added dropwise at 0°C. The resulting mixture was stirred for 1 hour at room temperature and under air. The resulting mixture was diluted with water (100 mL). The aqueous layer was extracted with CH₂Cl₂ (3 x 100 mL). The combined organic layers were concentrated under reduced pressure. The residue was purified by preparative TLC with PE / EA (40:1, v / v) to obtain a yellow oily substance, [(4-bromo-2-methoxyphenyl)methoxy](tert-butyl)dimethylsilane (2.56 g, 83.86%). 1H NMR (CD3OD, 400MHz) δ0.12(6H,s),0.97(9H,s),3.84(3H,s),4.70(2H,d),7.06–7.14(2H,m),7.31(1H,dt).

[0403] Step 2: At room temperature, add 8 mL of 1,4-dioxane containing [(4-bromo-2-methoxyphenyl)methoxy](tert-butyl)dimethylsilane (1 g, 3.02 mmol, 1 equivalent), methyl cyanoacetate (358.88 mg, 3.62 mmol, 1.20 equivalent), Pd(OAc)₂ (67.76 mg, 0.30 mmol, 0.1 equivalent), dppf (333.43 mg, 0.60 mmol, 0.2 equivalent), and t-BuOK (1.02 g, 9.05 mmol, 3 equivalent) to a 40 mL vial. Stir the resulting mixture overnight at 90 °C under a nitrogen atmosphere. Acidify the mixture to pH 5 with saturated NH₄Cl (aqueous solution). Stir the resulting mixture for 30 minutes at room temperature. Extract the aqueous layer with ethyl acetate (3 x 100 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by preparative TLC using PE / EA (4:1, v / v) to give methyl 2-(4-{[(tert-butyldimethylsilyl)oxy]methyl}-3-methoxyphenyl)-2-cyanoacetate I-34b (350 mg, 33.18%) as a yellow oil. 1 H NMR(DMSO-d6,400MHz)δ0.09(6H,d),0.92(9H,d),3.74(3H,d),3.81(3H,d),4.68(2H,s),5.62(1H,s),6.99–7.05(2H,m),7.42(1H,dd).

[0404] Step 3: At 0°C, add 5 mL of methanol containing nitrile I-34b (550 mg, 1.57 mmol, 1 equivalent), CoCl2·6H2O (973.46 mg, 4.09 mmol, 2.6 equivalent), and Boc2O (1.37 g, 6.30 mmol, 4 equivalent) to a 40 mL vial. At 0°C, add NaBH4 (476.25 mg, 12.59 mmol, 8 equivalent) to the above mixture. Stir the resulting mixture at room temperature for 2 hours. Quench the reaction with water at room temperature. Filter the resulting mixture and wash the filter cake with DCM (3 x 30 mL). Extract the filtrate with DCM (3 x 30 mL). Dry the combined organic layers with anhydrous Na2SO4, filter, and concentrate under reduced pressure. The residue was purified by preparative TLC using CH₂Cl₂ / methanol (40:1, v / v) to give methyl 3-[(tert-butoxycarbonyl)amino]-2-(4-{[(tert-butyldimethylsilyl)oxy]methyl}-3-methoxyphenyl)propionate (410 mg, 57.43%) as a yellow oil. LCMS: m / z (ESI), [M+Na) + =476.15. 1 H NMR(DMSO-d6,400MHz)δ0.08(6H,s),0.91(9H,s),1.34(9H,s),3.3(1H,m),3.41–3.54(1H,m),3 .60(3H,s),3.78(3H,s),3.85(1H,t),4.59–4.67(2H,m),6.83(2H,d),6.94(1H,t),7.29(1H,d).

[0405] Step 4: At room temperature, add 1 mL of water containing methyl 3-[(tert-butoxycarbonyl)amino]-2-(4-{[(tert-butyldimethylsilyl)oxy]methyl}-3-methoxyphenyl)propionate (390 mg, 0.86 mmol, 1 equivalent), THF (2 mL, 24.69 mmol, 28.71 equivalent), and LiOH·H₂O (72.15 mg, 1.72 mmol, 2.0 equivalent) to a 50 mL round-bottom flask. Stir the resulting mixture at room temperature for 1.5 hours. Purify the residue by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, methanol / water, 0% to 100% gradient, 30 min; detector, UV 220 nm. The resulting mixture was concentrated under vacuum to give 3-[(tert-butoxycarbonyl)amino]-2-(4-{[(tert-butyldimethylsilyl)oxy]methyl}-3-methoxyphenyl)propionic acid I-34 (160 mg, 42.33%) as a gray solid. LCMS: m / z (ESI), [M+Na) + =462.15.

[0406] Synthesis of intermediate I-35

[0407]

[0408] Step 1: At room temperature, dioxane (40 mL, 590.14 mmol, 27.74 equivalents) containing 3-bromo-5-methoxypyridine I-35a (4 g, 21.27 mmol, 1 equivalent) and methyl cyanoacetate (2.53 g, 25.53 mmol, 1.2 equivalents), Pd(OAc)₂ (477.62 mg, 2.13 mmol, 0.1 equivalent), dppf (2.35 g, 4.26 mmol, 0.2 equivalent), and t-BuOK (7.16 g, 63.82 mmol, 3 equivalents) was added to a 100 mL round-bottom flask. The mixture was stirred for 5 hours at 70 °C under a nitrogen atmosphere. The mixture was acidified to pH 6 with saturated NH₄Cl (aqueous solution). The mixture was extracted with ethyl acetate (3 x 60 mL). The combined organic layers were washed with DCM (3 x 3 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using CH2Cl2 / methanol (50:1, v / v) elution to give methyl 2-cyano-2-(5-methoxypyridin-3-yl)acetate (3.8 g, 86.63%) as a pale yellow solid. LCMS: m / z (ESI), [M+H] + =207.00.

[0409] Step 2: At 0°C and under a nitrogen atmosphere, NaBH4 (385.27 mg, 10.19 mmol, 3 equivalents) was added in portions to a stirred mixture of methyl 2-cyano-2-(5-methoxypyridin-3-yl)acetate (700 mg, 3.40 mmol, 1 equivalent) and CoCl2·6H2O (1.21 g, 5.09 mmol, 1.5 equivalents) in methanol (15 mL). The resulting mixture was stirred for 1 hour at room temperature under a nitrogen atmosphere. The reaction was quenched with water at room temperature. The resulting mixture was filtered, and the filter cake was washed with methanol (1 x 10 mL). The mixture was extracted with DCM (3 x 100 mL). The combined organic layers were washed with DCM (2 x 3 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC using CH₂Cl₂ / methanol (35:1, v / v) to give methyl 3-[(tert-butoxycarbonyl)amino]-2-(5-methoxypyridin-3-yl)propionate I-35b (250 mg, 23.73%) as a pale yellow solid. LCMS: m / z (ESI), [M+H] + =311.15.

[0410] Step 3: At room temperature, add 3 mL of water and 3 mL of methanol containing ester I-35b (80 mg, 0.26 mmol, 1 equivalent) and LiOH (12.35 mg, 0.52 mmol, 2 equivalents) to a 50 mL round-bottom flask. Stir the resulting mixture for 2 hours at room temperature under a nitrogen atmosphere. Acidify the mixture to pH 5 with concentrated HCl. Concentrate the resulting mixture under reduced pressure. Purify the residue by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, methanol / water, 28% to 32% gradient, 15 min; detector, UV 254 nm. Concentrate the resulting mixture under vacuum. This yields 3-[(tert-butoxycarbonyl)amino]-2-(5-methoxypyridin-3-yl)propionic acid I-35 (40 mg, 52.37%) as a grayish-white solid. LCMS: m / z (ESI), [M+H] + =297.05.

[0411] Synthesis of intermediate I-36

[0412]

[0413] Step 1: At room temperature, 4-bromo-1-methylpyridin-2-one I-36a (3 g, 15.96 mmol, 1 equivalent), methyl cyanoacetate (1.90 g, 19.15 mmol, 1.2 equivalent), Pd(OAc)₂ (358.21 mg, 1.60 mmol, 0.1 equivalent), Dppf (1.76 g, 3.19 mmol, 0.2 equivalent), t-BuOK (5.37 g, 47.87 mmol, 3 equivalent), and 1,4-dioxane (100 mL) were added to a 250 mL round-bottom flask. The resulting mixture was stirred for 1 hour at 70 °C under a nitrogen atmosphere. The reaction was quenched with water at room temperature. The mixture was acidified to pH 5 with HCl (1 M). The residue was diluted with water (100 mL). The resulting mixture was washed with 3 x 100 mL ethyl acetate. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in methanol (5 mL). The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, methanol / water, 0% to 0% gradient, 15 min; detector, UV 254 nm. The resulting mixture was concentrated under reduced pressure. This yielded methyl 2-cyano-2-(1-methyl-2-oxopyridin-4-yl)acetate (1.15 g, 34.95%) as a brown solid. LCMS: m / z (ESI), [M+H] + =207.05. 1H NMR (CD3OD, 400MHz) δ3.37(1H,s),3.45(3H,s),3.63(3H,s),6.75(1H,s),6.91(1H,s),7.23(1H,d).

[0414] Step 2: A solution of methyl 2-cyano-2-(1-methyl-2-oxopyridin-4-yl)acetate (1.1 g, 5.34 mmol, 1 equivalent) in methanol (15 mL) was treated with Boc₂O (4.66 g, 21.34 mmol, 4 equivalents) and CoCl₂·6H₂O (3.30 g, 13.87 mmol, 2.6 equivalents) for 5 min at 0 °C under air. Subsequently, NaBH₄ (1.61 g, 42.68 mmol, 8 equivalents) was added in portions at 0 °C. The resulting mixture was stirred for 15 h at 70 °C under air. The reaction was quenched by adding water (80 mL) at room temperature. The resulting mixture was filtered, and the filter cake was washed with DCM (3 x 10 mL). The aqueous layer was extracted with CH₂Cl₂ (3 x 80 mL). The resulting mixture was concentrated under reduced pressure. The residue was purified by preparative TLC with CH₂Cl₂ / methanol (18:1, v / v) to give methyl 3-[(tert-butoxycarbonyl)amino]-2-(1-methyl-2-oxopyridin-4-yl)propionate I-36b (445 mg, 26.88%) as a white oil. LCMS: m / z (ESI), [M+H] + =311.15.

[0415] Step 3: At room temperature, add ester I-36b (445 mg, 1.43 mmol, 1 equivalent), LiOH·H₂O (120.33 mg, 2.87 mmol, 2 equivalents), THF (3 mL), and H₂O (3 mL) to a 100 mL round-bottom flask. Stir the resulting mixture for 2 hours at room temperature under air atmosphere. Acidify the mixture to pH 5 with HCl (1 M). Concentrate the resulting mixture under reduced pressure. Dissolve the residue in methanol (5 mL). Purify the residue by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, methanol / water, 0% to 10% gradient, 30 min; detector, UV 220 nm. Concentrate the resulting mixture under reduced pressure. Dry the resulting liquid by lyophilization. This yielded 3-[(tert-butoxycarbonyl)amino]-2-(1-methyl-2-oxopyridin-4-yl)propionic acid I-36 (350 mg, 82.37%) as a grayish-white solid. LCMS: m / z (ESI), [M+H] + =297.15. 1H NMR (CD3OD, 400MHz) δ1.43(9H,d),3.44(2H,m),3.56(3H,d),3.69(1H,d),6.43(1H,d),6.50(1H,s),7.63(1H,d).

[0416] Synthesis of intermediate I-37

[0417]

[0418] Step 1: Sodium hydride (4 g, 166.68 mmol, 3.27 equivalents) was added to a stirred solution of 4-bromo-phenylacetonitrile I-37a (10 g, 51.01 mmol, 1 equivalent) in THF (120 mL) at 0 °C. The resulting mixture was stirred at 0 °C under air atmosphere for 1.5 h. Then, dimethyl carbonate (18.38 g, 204.03 mmol, 4 equivalents) was added to the above mixture at 0 °C. After the reaction, the resulting mixture was diluted with water (20 mL). The mixture was extracted with ethyl acetate (3 x 30 mL). The combined organic layers were dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC with PE / EA (2:1, v / v) to give methyl 2-(4-bromophenyl)-2-cyanoacetate (8.923 g, 68.85%) as a yellow oil. 1 HNMR(CDCl3,400MHz)δ3.84(3H,s),4.72(1H,s),7.35–7.39(2H,m),7.56–7.62(2H,m).

[0419] Step 2: At room temperature, add 70 mL of methanol containing methyl 2-(4-bromophenyl)-2-cyanoacetate (2 g, 7.87 mmol, 1 equivalent), Boc₂O (3.44 g, 15.74 mmol, 2 equivalents), and CoCl₂·6H₂O (3.75 g, 15.74 mmol, 2 equivalents) to a 250 mL round-bottom flask. Add NaBH₄ (1.79 g, 47.23 mmol, 6 equivalents) to the mixture at 0 °C. Stir the resulting mixture at 0 °C for 2 hours. Filter the mixture and wash the filter cake with DCM (4 x 50 mL). Extract the filtrate with CH₂Cl₂ (3 x 100 mL). Concentrate the combined organic layers under reduced pressure. The residue was purified by silica gel column chromatography using PE / EA (1:1, v / v) elution to give methyl 2-(4-bromophenyl)-3-[(tert-butoxycarbonyl)amino]propionate I-37b (1.9 g, 67.38%) as a yellow oil. LCMS: m / z (ESI), [M+H- t Bu] +=301.9 / 303.90. 1 H NMR (DMSO-d6, 400MHz) δ1.33(9H,s),3.26(1H,dt),3.42–3.53(1H,m),3.60(3H,s),3.85(1H,t),7.19–7.25(2H,m),7.51–7.57(2H,m).

[0420] Step 3: At room temperature, add 10 mL of DMF containing ester I-37b (1.9 g, 5.30 mmol, 1 equivalent), Zn(CN)₂ (0.62 g, 5.30 mmol, 1.0 equivalent), and Pd(PPh₃)₄ (0.61 g, 0.53 mmol, 0.1 equivalent) to a 40 mL vial. Stir the resulting mixture at 80 °C under a nitrogen atmosphere for 3 hours. Wash the residue with saturated NaHCO₃ (1 x 40 mL). Extract the resulting mixture with ethyl acetate (3 x 200 mL). Concentrate the combined organic layers under reduced pressure. Purify the residue by preparative TLC with PE / EA (1:1, v / v) to give methyl 3-[(tert-butoxycarbonyl)amino]-2-(4-cyanophenyl)propionate (1.0 g, 61.95%) as a yellow solid. LCMS: m / z (ESI), [M+H-tBu] + =248.95. 1 H NMR (DMSO-d6, 400MHz) δ1.31(9H,s),3.51(1H,ddd),3.62(3H,s),4.01(2H,dt),7.44–7.50(2H,m),7.82(2H,d).

[0421] Step 4: At room temperature, add 2.5 mL of water containing methyl 3-[(tert-butoxycarbonyl)amino]-2-(4-cyanophenyl)propionate (900 mg, 2.96 mmol, 1 equivalent), methanol (3.75 mL), and LiOH·H₂O (248.16 mg, 5.91 mmol, 2 equivalents) to a 20 mL vial. Stir the resulting mixture at 70 °C for 1.5 h. Acidify the mixture to pH 5 with HCl (aqueous solution). Purify the residue by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, methanol / water, 0% to 100% gradient, 30 min; detector, UV 254 nm. Concentrate the resulting mixture under vacuum to give 3-[(tert-butoxycarbonyl)amino]-2-(4-carbamoylphenyl)propionate I-37 (800 mg, 87.74%) as a white solid. LCMS:m / z(ESI),[M+H- t Bu] + =253.00.

[0422] Synthesis of intermediate I-38

[0423]

[0424] Step 1: At 80°C and under air, (Bu)3SnCH2OH (700 mg, 2.18 mmol, 1.5 equivalent) and Pd(PPh3)4 (84 mg, 0.073 mmol, 0.05 equivalent) were added to a solution of methyl 2-(4-bromophenyl)-2-[(tert-butoxycarbonyl)amino]acetate I-38a (500 mg, 1.45 mmol, 1 equivalent) in dioxane (30 mL) for 16 hours. The mixture was cooled to room temperature. The resulting mixture was quenched with water and extracted with ethyl acetate (3 x 30 mL). The combined organic layers were washed with saturated NaCl (3 x 30 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC using petroleum ether / ethyl acetate (3:1, v / v) elution to give 325 mg of methyl 2-[(tert-butoxycarbonyl)amino]-2-[4-(hydroxymethyl)phenyl]acetate I-38b (76%) as an orange oil. LCMS: m / z (ESI), [M+H] + =296.14. 1 H NMR (DMSO-d6, 400MHz) δ1.39(9H,s),3.62(3H,s),5.22-5.26(1H,d),7.35-7.38(2H,m),7.52-7.59(2H,m),7.83-7.86(1H,d).

[0425] Step 2: At room temperature, under a nitrogen atmosphere, a mixture of methyl 2-[(tert-butoxycarbonyl)amino]-2-[4-(hydroxymethyl)phenyl]acetate (390 mg, 1.30 mmol, 1 equivalent) and LiOH·H₂O (109 mg, 2.60 mmol, 2 equivalent) in THF (4 mL) and H₂O (1 mL) was dissolved for 5 hours. The resulting mixture was concentrated under reduced pressure. The residue was purified by preparative HPLC under the following conditions (mobile phase consisting of water containing 0.1% NH₄HCO₃ and acetonitrile containing 0.1% NH₄OH; using a constant gradient, mobile phase: 95% aqueous / 5% organic to 5% aqueous / 95% organic, 25 min; flow rate constant at 40 mL / min) to give 312 mg of [(tert-butoxycarbonyl)amino][4-(hydroxymethyl)phenyl]acetic acid I-38 (84%) as a yellow solid. LCMS:m / z(ESI),[M+H] + =282.13.

[0426] Synthesis of intermediate I-39

[0427]

[0428] Step 1: Add K₂CO₃ (2171.96 mg, 15.72 mmol, 1.20 equivalent) and polyoxymethylene (601.93 mg, 13.36 mmol, 1.02 equivalent) to a mixture of methyl 2-(3-bromophenyl)acetate I-39a (3.0 g, 13.10 mmol, 1 equivalent) in DMF (4.0 mL). Stir the resulting mixture at 70 °C under a nitrogen atmosphere for 2 hours. After cooling to room temperature, dilute the mixture with water (40 mL). Extract the mixture with ethyl acetate (3 x 40 mL). Wash the combined organic layers with water (3 x 10 mL) and dry over anhydrous Na₂SO₄. Filter and concentrate the filtrate under reduced pressure. The residue was purified by preparative TLC with PE / EA (3:1, v / v) to give methyl 2-(3-bromophenyl)prop-2-enoate (850 mg, 26.92%) as a yellow oil. 1 H NMR(DMSO-d6,400MHz)δ3.76(3H,s),6.12(1H,d),6.33(1H,d),7.35(1H,t),7.44(1H,ddd),7.57(1H,ddd),7.64(1H,t).

[0429] Step 2: Add 1-methylpiperazine (1661.86 mg, 16.59 mmol, 5 equivalents) to a mixture of methyl 2-(3-bromophenyl)prop-2-enoate (800 mg, 3.32 mmol, 1 equivalent) in DMF (5 mL). Stir the resulting mixture at 25 °C under a nitrogen atmosphere for 2 hours. Dilute the reaction mixture with EA (20 mL). Wash the residue with saturated NaHCO3 (2 x 10 mL), then with water (2 x 20 mL), and finally with saturated NaCl (3 x 20 mL). Dry the resulting organic layer with Na2SO4. Concentrate the resulting mixture under reduced pressure. Purify the residue by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water, 10% to 50% gradient, 10 min; detector, UV 254 nm, 65% hold for 30 min. After evaporation of water, methyl 2-(3-bromophenyl)-3-(4-methylpiperazin-1-yl)propionate I-39b (740 mg, 65.35%) was obtained as a yellow oil. LCMS: m / z (ESI), [M+H] + =341.00,343.00.

[0430] Step 3: At room temperature and under a nitrogen atmosphere, Pd(PPh3)4 (152.39 mg, 0.13 mmol, 0.1 equivalent) was added in portions to a stirred mixture of methylpiperazine I-39b (740 mg, 1.32 mmol, 1 equivalent) and Zn(CN)2 (154.84 mg, 1.32 mmol, 1.0 equivalent) in DMF (3.0 mL). The resulting mixture was stirred at 80 °C under a nitrogen atmosphere for 2.0 h. The mixture was cooled to room temperature. The resulting mixture was diluted with water (20 mL). The resulting mixture was extracted with ethyl acetate (3 x 30 mL). The combined organic layers were washed with water (3 x 10 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water, 10% to 50% gradient, 10 min; detector, UV 254 nm, 28% hold for 30 min. After solvent evaporation, methyl 2-(3-cyanophenyl)-3-(4-methylpiperazin-1-yl)propionate (400 mg, 52.50%) was obtained as a yellow oil. LCMS: m / z (ESI), [M+H] + =288.05.

[0431] Step 4: At room temperature, under a nitrogen atmosphere, water (2.0 mL) containing LiOH·H₂O (175.22 mg, 4.18 mmol, 3.0 equivalent) was added dropwise to a stirred mixture of methyl 2-(3-cyanophenyl)-3-(4-methylpiperazin-1-yl)propionate (400 mg, 1.39 mmol, 1 equivalent) in THF (8.0 mL). The resulting mixture was stirred at 25 °C under a nitrogen atmosphere for 4.0 h. After the reaction, the resulting mixture was concentrated under vacuum. The mixture was diluted with methanol (4.0 mL). The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water, 0% to 50% gradient, 10 min; detector, UV 254 nm, 9.8% hold for 20 min, to give 2-(3-cyanophenyl)-3-(4-methylpiperazin-1-yl)propionic acid I-39 (240 mg, 63.08%) as a white solid. LCMS: m / z (ESI), [M+H) + =274.10.

[0432] Synthesis of intermediate I-40

[0433]

[0434] Step 1: At room temperature, add 3-bromo-5-methoxypyridine I-40a (8 g, 42.55 mmol, 1 equivalent) and THF (40 mL) to a 100 mL three-necked round-bottom flask. At -10 °C under a nitrogen atmosphere, add a 1 M solution of isopropyl magnesium bromide in THF (17.61 mL, 51.06 mmol, 1.2 equivalent). At -10 °C under a nitrogen atmosphere, stir the mixture for 30 minutes. At -10 °C, add ethyl 2-oxobutyrate (6.64 g, 51.06 mmol, 1.2 equivalent) to the above mixture. At -8 °C under a nitrogen atmosphere, stir the resulting mixture for 3 hours. Quench the reaction by adding saturated NH4Cl (aqueous solution) (50 mL) at 0 °C. Extract the resulting mixture with ethyl acetate (3 x 30 mL). The combined organic layers were dried over anhydrous Na₂SO₄, filtered, and evaporated to obtain a crude oil. The residue was purified by silica gel column chromatography using PE / EA (1:1, v / v) elution to give ethyl 2-(5-methoxypyridin-3-yl)-2-oxoethyl acetate I-40b (4.6 g, 45.48%) as a yellow oil. LCMS: m / z (ESI), [M+H] + =210.05. 1 H NMR (CD3OD, 400MHz) δ1.23(3H,t),3.91(3H,s),4.15–4.26(2H,m),7.61(1H,dd),8.23(1H,d),8.37(1H,d).

[0435] Step 2: At 0 °C, ester I-40b (2 g, 9.56 mmol, 1 equivalent), hydroxylamine hydrochloride (0.86 g, 12.43 mmol, 1.3 equivalent), pyridine (0.83 g, 10.52 mmol, 1.1 equivalent), and methanol (10 mL) were added to a 50 mL round-bottom flask. The resulting mixture was stirred for 6 hours at room temperature. The mixture was diluted with CH2Cl2 (40 mL). The mixture was washed with 1 x 40 mL HCl (0.5 M), 1 x 40 mL water, and 1 x 40 mL saturated NaCl (aqueous solution). The organic layer was concentrated under vacuum. The precipitated solid in the aqueous phase was collected by filtration and washed with water (3 x 10 mL). This gave ethyl acetate (2E)-2-(N-hydroxyimino)-2-(5-methoxypyridin-3-yl) as a white solid (1.78 g, 83.04%). LCMS: m / z (ESI), [M+H] + =225.15.

[0436] Step 3: At room temperature, ethyl 2-(N-hydroxyimino)-2-(5-methoxypyridin-3-yl)acetate (1.7 g, 7.58 mmol, 1 equivalent), Pd / C (1.86 g, 17.44 mmol, 2.3 equivalent), and methanol (100 mL) were added to a 50 mL round-bottom flask. The mixture was stirred for 6 hours at room temperature under a hydrogen atmosphere. The mixture was filtered, and the filter cake was washed with methanol (6 x 30 mL). The filtrate was concentrated under reduced pressure. This yielded ethyl 2-amino-2-(5-methoxypyridin-3-yl)acetate I-40c (1.6 g, 87.33%) as a yellow oil. LCMS: m / z (ESI), [M+H] + =211.10. 1 H NMR (CD3OD, 400MHz) δ1.22(3H,t),4.19(2H,m),4.69(1H,d),4.88(3H,s),7.50(1H,dq),8.20(2H,t).

[0437] Step 4: At room temperature, amine I-40c (1.6 g, 7.61 mmol, 1 equivalent), triethylamine (3.85 g, 38.06 mmol, 5 equivalent), Boc2O (3.32 g, 15.22 mmol, 2 equivalent), and DCM (5 mL) were added to a 50 mL round-bottom flask. The resulting mixture was stirred for 2 hours at room temperature. The mixture was quenched with water (50 mL) and extracted with DCM (2 x 50 mL). The combined organic layers were washed with saturated brine (1 x 50 mL). The organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC with CH2Cl2 / methanol (25:1, v / v) to give ethyl acetate 2-[(tert-butoxycarbonyl)amino]-2-(5-methoxypyridin-3-yl) as a yellow oil (1.38 g, 49.19%). LCMS:m / z(ESI),[M+H] + =311.20. 1 H NMR (CD3OD, 400MHz) δ1.19–1.29(3H,m),1.47(9H,s),3.91(3H,s),4.10–4.31(2H,m),7.46(1H,q),8.10–8.27(2H,m).

[0438] Step 5: A solution of ethyl 2-[(tert-butoxycarbonyl)amino]-2-(5-methoxypyridin-3-yl)acetate (1.38 g, 4.45 mmol, 1 equivalent) and LiOH (0.21 g, 8.89 mmol, 2 equivalents) in THF (13 mL) and water (13 mL) was stirred for 6 hours at 60 °C. The mixture / residue was acidified to pH 5 with concentrated HCl. The precipitate was collected by filtration and washed with methanol (2 x 10 mL). The filtrate was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, methanol / water, 0 to 50 gradient, 30 min; detector, UV 254 nm. This yielded [(tert-butoxycarbonyl)amino](5-hydroxypyridin-3-yl)acetic acid I-40 (500 mg, 41.41%) as a white solid. LCMS: m / z (ESI), [M+H] + =283.15.

[0439] Synthesis of intermediate I-41

[0440]

[0441] Methyl 3-[(tert-butoxycarbonyl)amino]-2-(3-cyanophenyl)propionate I-3d (700 mg, 2.30 mmol, 1 equivalent) and LiOH·H₂O (220.34 mg, 9.20 mmol, 4 equivalent) were stirred for 4.5 h in MeOH (8 mL) and H₂O (4 mL) at 70 °C under air atmosphere. The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water, 10% to 100% gradient, 50 min; detector, UV 220 nm. This yielded 3-[(tert-butoxycarbonyl)amino]-2-(3-carbamoylphenyl)propionate I-41 (330 mg, 46.53%) as a yellow solid. LCMS: m / z (ESI), [MH] + =307.05. 1 H NMR (CD3OD, 400MHz) δ1.40(9H,s),3.50(2H,m),3.73(1H,t),7.40(1H,t),7.56(1H,d),7.73(1H,d),7.86(1H,t).

[0442] Synthesis of intermediate I-42

[0443]

[0444] Step 1: Under a nitrogen atmosphere at room temperature, Boc2O (1.92 g, 8.81 mmol, 1.2 equivalent) was added to a stirred mixture of 2-amino-1-(pyridin-3-yl)ethyl ketone I-42a (1.0 g, 7.35 mmol, 1 equivalent) and Et3N (2.23 g, 22.03 mmol, 3 equivalent) in a DCM (20 mL). The resulting mixture was stirred for 3 hours at room temperature. After the reaction, the resulting mixture was diluted with water (50 mL) and extracted with ethyl acetate (3 x 30 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC with PE / EA (1:1, v / v) to give N-[2-oxo-2-(pyridin-3-yl)ethyl]carbamate tert-butyl ester (650 mg, 37.46%) as a yellow solid. LCMS:[M+H] + =237.05. 1 H NMR (CDCl3, 400MHz) δ1.52(9H,s),4.69(2H,d),5.50(1H,s),7.48(1H,ddd),8.25(1H,dt),8.85(1H,dd),9.19(1H,m).

[0445] Step 2: At 0°C and under a nitrogen atmosphere, pyridine (642.79 mg, 8.13 mmol, 3 equivalents) was added dropwise to a stirred mixture of N-[2-oxo-2-(pyridin-3-yl)ethyl]carbamate (640 mg, 2.71 mmol, 1 equivalent) and hydroxylamine hydrochloride (564.70 mg, 8.13 mmol, 3 equivalents) in methanol (10 mL). The resulting mixture was stirred overnight at room temperature. After the reaction, the mixture was quenched with water (20 mL). The mixture was extracted with ethyl acetate (3 x 20 mL). The combined organic layers were dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure to give N-[2-(hydroxyimino)-2-(pyridin-3-yl)ethyl]carbamate tert-butyl I-42b (400 mg, 58.77%) as a yellow oil. LCMS:m / z(ESI),[M+H] + =252.00.

[0446] Step 3: At room temperature and under a nitrogen atmosphere, Zn (362.95 mg, 5.55 mmol, 3 equivalents) was added in portions to a stirred mixture of hydroxyimine I-42b (465 mg, 1.85 mmol, 1 equivalent) and formic acid (170.34 mg, 3.70 mmol, 2 equivalents) in methanol (4.0 mL). The resulting mixture was stirred at room temperature for 3 hours. The desired product was detected by LCMS. After the reaction, the resulting mixture was concentrated under vacuum. Then, it was diluted with methanol (4.0 mL). The residue was alkalized to pH 9 with K2CO3. The precipitate solid was filtered off and washed with DCM / methanol (10:1, v / v). The filtrate was evaporated and purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water, 10% to 50% gradient, 10 min; detector, UV 220 nm, 45% hold for 20 min. After evaporation of the solvent, N-[2-amino-2-(pyridin-3-yl)ethyl]carbamate tert-butyl ester I-42 (287 mg, 65.36%) was obtained as a yellow oil. LCMS: m / z (ESI), [M+H] + =238.10. 1 H NMR(DMSO-d6,400MHz)δ1.32(9H,s),2.05(2H,s),3.06(2H,m),3.90(1H,t),6.86(1H,t),7.32(1H,dd),7.73(1H,dt),8.41(1H,dd),8.49(1H,d).

[0447] Synthesis of intermediate I-43

[0448]

[0449] Step 1: At room temperature, add 30 mL of MeCN containing [(tert-butoxycarbonyl)amino](3-methoxyphenyl)acetic acid I-43a (3 g, 10.66 mmol, 1 equivalent) and 1-methyl-piperazine (1.28 g, 12.80 mmol, 1.2 equivalent), TCFH (4.49 g, 16.00 mmol, 1.5 equivalent), and NMI (4.38 g, 53.32 mmol, 5 equivalent) to a 250 mL round-bottom flask. Stir the resulting mixture for 3 hours at room temperature under a nitrogen atmosphere. The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, methanol / water, 0% to 70% gradient, 40 min; detector, UV 254 nm, to obtain N-[1-(3-methoxyphenyl)-2-(4-methylpiperazin-1-yl)-2-oxoethyl]carbamate tert-butyl ester I-43b (2.5 g, 64.50%) as a yellow oil. LCMS: m / z (ESI), [M+H]+ =364.20.

[0450] Step 2: At room temperature, add 4 mL of DCM containing amide I-43b (2.2 g, 6.05 mmol, 1 equivalent) and TFA (4 mL, 53.85 mmol, 8.90 equivalent) to a 50 mL round-bottom flask. Stir the resulting mixture for 15 hours at room temperature under a nitrogen atmosphere. Concentrate the mixture under reduced pressure. Alkalinize the residue to pH 9 with NH3·H2O. Purify the residue by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, methanol / water (5 mmol / L NH4HCO3), 10% to 50% gradient, 10 min; detector, UV 254 nm, to obtain 2-amino-2-(3-methoxyphenyl)-1-(4-methylpiperazin-1-yl)acetone I-43c (1.5 g, 94.10%) as a yellow oil. LCMS: m / z (ESI), [M+H- t Bu] + =264.05. 1 HNMR (CD3OD, 400MHz) δ2.62(1H,d),2.79(3H,s),3.16(3H,d),3.44(1H,s),3.85(5H,s),4.03(1H,s),5.55(1H,s),7.03–7.18(3H,m),7.45(1H,t).

[0451] Step 3: At room temperature, add amine I-43c (1.5 g, 5.70 mmol, 1 equivalent) and BH3.C4H8O (20 mL, 208.98 mmol, 36.69 equivalent) to a 40 mL vial. Stir the resulting mixture at 70 °C under a nitrogen atmosphere for 24 hours. Cool the mixture to room temperature. Quench the reaction by adding methanol (20 mL) at 0 °C. Then, stir the resulting mixture for 2 hours and concentrate under reduced pressure. The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, methanol / water, 0% to 50% gradient, 40 min; detector, UV 220 nm, to give 1-(3-methoxyphenyl)-2-(4-methylpiperazin-1-yl)ethylamine I-43 (500 mg, 35.20%) as a colorless oil. LCMS: m / z (ESI), [M+H- t Bu] + =250.05.

[0452] Synthesis of intermediate I-44

[0453]

[0454] Step 1: At room temperature, under a nitrogen atmosphere, a mixture of tribromotetrabutylamine (6.42 g, 13.32 mmol, 1 equivalent) and 1-[4-(hydroxymethyl)phenyl]acetone I-44a (2 g, 13.32 mmol, 1 equivalent) in ACN (100 mL) and acetone (10 mL) was stirred for 1 hour. The resulting mixture was concentrated under reduced pressure. The resulting mixture was diluted with water (10 mL). The resulting mixture was extracted with MTBE (2 x 20 mL). The combined organic layers were washed with water (7 x 20 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure to give 2-bromo-1-[4-(hydroxymethyl)phenyl]acetone I-44b (2 g, 65.56%) as a yellow oil. The resulting mixture was used directly in the next step.

[0455] Step 2: At room temperature, under a nitrogen atmosphere, a mixture of methanol (6.24 mL, 43.66 mmol, 5 equivalents) containing NH3 (g) and THF (50 mL) containing bromide I-44b (2 g, 8.73 mmol, 1 equivalent) was stirred for 2 hours. The resulting mixture was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water (0.1% NH3·H2O), 0% to 50% gradient, 60 min; detector, UV 254 nm, to give 2-amino-1-[4-(hydroxymethyl)phenyl]acetone I-44c (1 g, 69.34%) as a colorless oil. 1 HNMR (400MHz, DMSO-d6) δ2.56(2H,d),4.62(2H,s),5.43(1H,s),7.53(2H,d),8.00(2H,d),8.21(2H,s).

[0456] Step 3: At room temperature and under a nitrogen atmosphere, a mixture of Boc2O (1.32 g, 6.05 mmol, 1 equivalent), triethylamine (1.84 g, 18.16 mmol, 3 equivalents), and amine I-44c (1 g, 6.05 mmol, 1 equivalent) in DCM (20 mL) was stirred for 2 hours. The resulting mixture was concentrated under reduced pressure. The residue was purified by preparative TLC with water PE / EA (1:2, v / v) to give N-{2-[4-(hydroxymethyl)phenyl]-2-oxoethyl} tert-butyl carbamate I-44d (500 mg, 31.13%) as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ1.40(9H,s),4.43(2H,d),4.59(2H,s),5.38(1H,s),7.05(1H,t),7.47(2H,d),7.89–8.00(2H,m).

[0457] Step 4: At 80°C under a nitrogen atmosphere, a mixture of NaBH3CN (278.31 mg, 4.43 mmol, 2.5 equivalents), NH4OAc (682.77 mg, 8.86 mmol, 5 equivalents), and ketone I-44d (470 mg, 1.77 mmol, 1 equivalent) in methanol (10 mL) was stirred for 2 hours. The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, methanol / water (10 mmol / L NH4HCO3), 0% to 100% gradient, 60 min; detector, UV 254 nm, to obtain N-{2-amino-2-[4-(hydroxymethyl)phenyl]ethyl}carbamate tert-butyl ester I-44 (110 mg, 23.31%) as a yellow oil. 1 H NMR (400MHz, DMSO-d6) δ1.41(9H,s),4.43(2H,d),4.59(3H,d),5.37(1H,t),7.04(1H,t),7.47(2H,d),7.94(2H,s).

[0458] Synthesis of intermediate I-45

[0459]

[0460] Step 1: A solution of methyl pyruvate I-45a (5 g, 48.98 mmol, 1 equivalent) and 4-methylbenzenesulfonylhydrazine (10.03 g, 53.88 mmol, 1.1 equivalent) in 1,4-dioxane (30 mL) was stirred for 2 hours at 70 °C under a nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was purified by grinding with methanol (20 mL) to give methyl [N'-(4-methylbenzenesulfonyl)hydrazine carbonyl]formate I-45b (12.8 g, 95.98%) as a grayish-white solid. LCMS: m / z (ESI), [M+NH4) + =288.05.

[0461] Step 2: At room temperature, add 100 mL of dioxane containing ester I-45b (5000 mg, 18.36 mmol, 1 equivalent), Pd2(dba)3 (840.8 mg, 0.92 mmol, 0.05 equivalent), XPhos (1750.9 mg, 3.67 mmol, 0.2 equivalent), (tert-butoxy)lithium (3528.2 mg, 44.06 mmol, 2.4 equivalent), and 3-bromo-5-methoxypyridine (3452.8 mg, 18.36 mmol, 1.0 equivalent) to a 40 mL vial. Stir the resulting mixture at 110 °C under a nitrogen atmosphere for 20 minutes. Quench the reaction with saturated NH4Cl (aqueous solution) at 0 °C. Quench the resulting mixture with water (100 mL) and extract with ethyl acetate (3 x 100 mL). The combined organic layers were dried over anhydrous Na₂SO₄, filtered, and evaporated to obtain a crude solid. The residue was purified by preparative TLC using CH₂Cl₂ / methanol (45:1, v / v) to give methyl 2-(5-methoxypyridin-3-yl)prop-2-enoate I-45c (1500 mg, 42.28%) as a yellow oil. LCMS: m / z (ESI), [M+H] + =193.95.

[0462] Step 3: At room temperature, add 3 mL of DMF containing acrylate I-45c (1.5 g, 7.76 mmol, 1 equivalent), triethylamine (2.36 g, 23.29 mmol, 3 equivalents), and 1-methylpiperazine (777.65 mg, 7.76 mmol, 1 equivalent) to a 50 mL round-bottom flask. Stir the resulting mixture for 1.5 hours at room temperature under a nitrogen atmosphere. Concentrate the resulting mixture under vacuum. Purify the residue by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, methanol / water, 0% to 100% gradient, 30 min; detector, UV 254 nm. Concentrate the resulting mixture under vacuum to give methyl 2-(5-methoxypyridin-3-yl)-3-(4-methylpiperazin-1-yl)propionate I-45d (900 mg, 39.51%) as a white oil. LCMS:m / z(ESI),[M+H] + =294.10.

[0463] Step 4: At room temperature, add 2 mL of water containing methylpiperazine I-45d (900 mg, 3.07 mmol, 1 equivalent), THF (2 mL), and LiOH·H₂O (257.45 mg, 6.14 mmol, 2 equivalents) to a 50 mL round-bottom flask. Stir the resulting mixture at room temperature for 2 hours. Acidify the mixture to pH 5 with HCl (aqueous solution). Purify the residue by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, methanol / water, 0% to 100% gradient, 30 min; detector, UV 254 nm. Concentrate the resulting mixture under vacuum to give 2-(5-methoxypyridin-3-yl)-3-(4-methylpiperazin-1-yl)propionic acid I-45 (330 mg, 38.51%) as a white solid. LCMS: m / z (ESI), [M+H] + =280.10. 1 H NMR (DMSO-d6, 400MHz) δ2.70(3H,s),3.05(3H,dd),3.36(7H,s),3.84(3H,s),3.94(1H,t),7.34(1H,dd),8.16(1H,d),8.22(1H,d).

[0464] Synthesis of intermediate I-46

[0465]

[0466] At room temperature, 20.00 mL of methanol containing tert-butyl N-(3-oxo-3-phenylpropyl)carbamate I-46a (400 mg, 1.60 mmol, 1 equivalent), sodium deuterated cyanoborocyanate (403.30 mg, 6.42 mmol, 4 equivalents), and NH4OAc (1.24 g, 16.04 mmol, 10 equivalents) was added to a 20 mL vial. The resulting mixture was stirred overnight at 60 °C under a nitrogen atmosphere. The mixture was alkalized to pH 9 with NaOH (2 mol / L). The mixture was extracted with ethyl acetate (3 x 100 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by preparative TLC using CH₂Cl₂ / methanol (15:1, v / v) to give N-(3-amino-3-phenylpropyl)carbamate tert-butyl ester I-46 (250 mg, 62.24%) as a yellow oil. LCMS: m / z (ESI), [M+H] + =251.15. 1H NMR(DMSO-d6,400MHz)δ1.37(9H,s),1.66–1.80(2H,m),2.89(2H,p),3.89(1H,t),7.25(1H,ddt),7.29–7.41(4H,m).

[0467] Synthesis of intermediate I-47

[0468]

[0469] Step 1: At 0°C and under air, magnesium bromo-(3-methoxyphenyl)methyl methoxyphenyl ... The residue was purified by silica gel column chromatography using CH2Cl2 / methanol (50:1, v / v) elution to give 4-(3-methoxybenzoyl)-1-methylpiperidine I-47b (900 mg, 71.85%) as a white solid. LCMS: m / z (ESI), [M+H] + =234.10. 1 HNMR(CDCl3,400MHz)δ1.90(4H,m),2.14(2H,m),2.34(3H,s),2.96(2H,dd),3.22(1H,tt),3.87(3H,s),7.12(1H,ddd),7.39(1H,t),7.50(2H,m).

[0470] Step 2: At 0°C and under air, NaBH3CN (948.08 mg, 15.09 mmol, 4 equivalents) and CH3COONH4 (3488.89 mg, 45.26 mmol, 12 equivalents) were added in portions to a stirred mixture of methylpiperidine I-47b (880 mg, 3.77 mmol, 1 equivalent) in methanol (10 mL). The resulting mixture was stirred overnight at 70°C and under air. After cooling to room temperature, the reaction was quenched by adding saturated NH4Cl (aqueous solution) (30 mL) at 0°C. The resulting mixture was extracted with CH2Cl2 (3 x 40 mL). The combined organic layers were washed with water (3 x 10 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water, 10% to 50% gradient, 10 min; detector, UV 254 nm, 40% hold for 25 min, to give 1-(3-methoxyphenyl)-1-(1-methylpiperidin-4-yl)methylamine I-47 (340 mg, 38.47%) as a yellow solid. LCMS: m / z (ESI), [M+H) + =235.15.

[0471] Synthesis of intermediate I-48

[0472]

[0473] Step 1: At room temperature, add 10.00 mL of THF containing N-methoxy-N,1-dimethylpiperidine-4-carboxamide I-47a (2 g, 10.74 mmol, 1 equivalent) and magnesium bromo(cyclohexyl)magnesium (21.48 mL, 21.48 mmol, 2 equivalents) to a 40 mL vial. Stir the resulting mixture overnight at room temperature under a nitrogen atmosphere. Quench the reaction with water at room temperature. Extract the resulting mixture with ethyl acetate (3 x 20 mL). Concentrate the combined organic layers under reduced pressure. Purify the residue by silica gel column chromatography using CH2Cl2 / methanol (20:1, v / v) elution to give 4-cyclohexanecarbonyl-1-methylpiperidine I-48b (270 mg, 12.01%) as a yellow oil. LCMS: m / z (ESI), [M+H] + =210.20. 1 H NMR(DMSO-d6,400MHz)δ1.08–1.32(5H,m),1.35–1.48(2H,m),1.56–1.78(7H ,m),1.86(2H,td),2.14(3H,s),2.46(1H,ddt),2.56(1H,td),2.75(2H,dt).

[0474] Step 2: At room temperature, add 5 mL of methanol containing methylpiperidine I-48b (260 mg, 1.24 mmol, 1 equivalent), NH4OAc (1.34 g, 17.39 mmol, 14 equivalents), and sodium deuterated cyanoboronide (312.21 mg, 4.97 mmol, 4 equivalents) to a 20 mL vial. Stir the resulting mixture overnight at 60 °C under a nitrogen atmosphere. Purify the residue by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, water / MEOH, 0% to 100% gradient, 30 min; detector, UV 254 nm. Concentrate the resulting mixture under vacuum to give 1-cyclohexyl-1-(1-methylpiperidin-4-yl)methylamine I-48 (180 mg, 68.89%) as a brown oil. LCMS: m / z (ESI), [M+H] + =211.20. 1 H NMR(DMSO-d6,400MHz)δ0.96(1H,qd),1.04–1.12(1H,m),1.17(4H,ddt),1.22–1.34(3 H,m),1.44(2H,ddd),1.54–1.79(7H,m),2.08(1H,d),2.11(3H,s),2.70–2.81(2H,m).

[0475] Synthesis of intermediate I-49

[0476]

[0477] Step 1: At room temperature, 3-bromo-7-nitro-1H-indole I-49a (20 g, 82.97 mmol, 1 equivalent), 4-(4,4,5,5-tetramethyl-1,3,2-dioxoboron-2-yl)pyrazole-1-carboxylic acid tert-butyl ester (26.85 g, 91.27 mmol, 1.1 equivalent), K₂CO₃ (34.40 g, 248.92 mmol, 3 equivalent), Pd(dppf)Cl₂ (3.04 g, 4.15 mmol, 0.05 equivalent), 1,4-dioxane (80 mL, 0.023 mmol), and water (20 mL, 27.76 mmol) were added to a 250 mL round-bottom flask. The resulting mixture was stirred for 2 hours at 80 °C under a nitrogen atmosphere. The residue was purified by grinding with water (30 mL). This yielded a crude solid (26 g). The coarse solid was ground with methanol (50 mL), yielding tert-butyl 4-(7-nitro-1H-indol-3-yl)pyrazole-1-carboxylate I-49b (17 g, 39.31%) as a brownish-yellow solid. LCMS: m / z (ESI), [M-Boc] + =229.05.

[0478] Step 2: At 0°C, add nitro-indole I-49b (2.5 g, 7.61 mmol, 1 equivalent) and Zn (3.98 g, 60.91 mmol, 8 equivalents) to a 100 mL round-bottom flask. At 0°C, add a solution of NH4Cl (1.79 g, 33.51 mmol, 5 equivalents) in 10 mL of water to the above mixture. At 0°C, stir the resulting mixture under a nitrogen atmosphere for 1 hour. Filter the resulting mixture and wash the filter cake with ethyl acetate (2 x 20 mL). Dilute the filtrate with water (50 mL) and extract the residue with EA (2 x 50 mL). Dry the combined organic layers over anhydrous Na2SO4, filter, and evaporate to give a crude solid. The residue was purified by preparative TLC with CH2Cl2 / methanol (25:1, v / v) to give tert-butyl 4-(7-amino-1H-indol-3-yl)pyrazole-1-carboxylate I-49 (1 g, 44.02%) as a brown solid. LCMS: m / z (ESI), [M- t Bu] + =243.10.

[0479] Synthesis of intermediate I-50

[0480]

[0481] Step 1: At 90°C and under nitrogen, dioxane (5.00 mL) containing tert-butyl 3-bromo-7-nitro-indole-1-carboxylate I-50a (219 mg, 641.93 μmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxorane-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole (249.81 mg, 770.32 μmol), potassium carbonate (266.16 mg, 1.93 mmol), and Pd(dppf)Cl2 (52.42 mg, 64.19 μmol) and water (1.00 mL) was stirred for 16 hours. The solvent was removed under reduced pressure. The residue was purified by silica gel column chromatography with a methanol / DCM gradient elution from 0% to 10%. The pure fraction was evaporated to dryness to give trimethyl-[2-[[4-(7-nitro-1H-indol-3-yl)pyrazol-1-yl]methoxy]ethyl]silane I-50b (200 mg, 87% yield) as a brown solid. LCMS: m / z (ESI), [M+H] + =359.3.

[0482] Step 2: Iron powder (155.79 mg, 2.79 mmol) was added to a mixture of nitro-indole I-50b (200 mg, 557.93 μmol) and ammonium hydrochloride (298.45 mg, 5.58 mmol) in ethanol (10 mL) and water (10 mL). The reaction mixture was then stirred and heated at 80 °C for 3 hours. The reaction mixture was filtered through diatomaceous earth and washed with ethanol (3 x 10 mL), and the filtrate was concentrated under reduced pressure. The residue was dissolved in 50 mL of ethyl acetate, filtered, and the filtrate was concentrated under reduced pressure to give 3-[1-(2-trimethylsilylethoxymethyl)pyrazol-4-yl]-1H-indole-7-amine I-50 (160 mg, 87% yield) as a brown solid. LCMS: m / z (ESI), [M+H] + =329.3.

[0483] Synthesis of intermediate I-51

[0484]

[0485] Step 1: At -10°C, methyl 1H-indole-7-carboxylate I-51a (5 g, 28.54 mmol, 1 equivalent) and DCE (50 mL) were added to a 50 mL three-necked round-bottom flask. Under a nitrogen atmosphere, NBS (6.10 g, 34.25 mmol, 1.2 equivalent) was added in portions to the stirred mixture at -10°C. The resulting mixture was quenched with water (80 mL) and extracted with CH2Cl2 (3 x 50 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using PE / EA (5:1, v / v) elution to give methyl 3-bromo-1H-indole-7-carboxylate I-51b (5.1 g, 68.71%) as a white solid. LCMS: m / z (ESI), [M+H] + =253.95. 1 H NMR (400MHz, DMSO-d6) δ3.95(3H,s),7.27(1H,t),7.60(1H,d),7.74(1H,dt),7.88(1H,dd),11.51(1H,s).

[0486] Step 2: At 80°C under a nitrogen atmosphere, a solution of bromide I-51b (2 g, 7.87 mmol, 1 equivalent), Pd(dppf)Cl2.CH2Cl2 (0.64 g, 0.79 mmol, 0.1 equivalent), K2CO3 (3.26 g, 23.61 mmol, 3 equivalent), and 4-(4,4,5,5-tetramethyl-1,3,2-dioxoboron-2-yl)-1H-pyrazole (1.53 g, 7.87 mmol, 1 equivalent) in 1,4-dioxane (4 mL) and water (1 mL) was stirred for 12 hours. The resulting mixture was diluted with water (30 mL), extracted with ethyl acetate (3 x 30 mL), and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using CH2Cl2 / methanol (5:1, v / v) elution to obtain methyl 3-[1-(tert-butoxycarbonyl)pyrazol-4-yl]-1H-indole-7-carboxylate I-51c (810 mg, 19.41%), a yellow oil. LCMS: m / z (ESI), [M+H] + =242.00.

[0487] Step 3: At room temperature, ester I-51c (390 mg, 1.14 mmol, 1 equivalent), LiOH (136.81 mg, 5.71 mmol, 5 equivalent), THF (2.5 mL), and water (2.5 mL) were added to a 50 mL round-bottom flask. The resulting mixture was stirred for 8 hours at room temperature. The mixture was acidified to pH 5 with HCl (1 M). The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, methanol / water, 0% to 20% gradient, 30 min; detector, UV 254 nm. This yielded 3-(1H-pyrazol-4-yl)-1H-indole-7-carboxylic acid I-51 (60 mg, 13.98%) as a yellow solid. LCMS: m / z (ESI), [M+H] + =242.10.

[0488] Synthesis of intermediate I-52

[0489]

[0490] Step 1: To a mixture of methyl 2-amino-2-(3-bromophenyl)acetate I-52a (2.0 g, 8.19 mmol, 1 equivalent) in DCM (30 mL), add (Boc)₂O (3576.56 mg, 16.39 mmol, 2 equivalents) and TEA (2487.46 mg, 24.58 mmol, 3 equivalents). Stir the resulting mixture at room temperature under air atmosphere for 4 hours. After the reaction, concentrate the resulting mixture under vacuum and dissolve it in DMF (4 mL). The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, MeOH / water, 10% to 100% gradient, 10 min; detector, UV 254 nm, 90% hold for 30 min, to give methyl 2-(3-bromophenyl)-2-[(tert-butoxycarbonyl)amino]acetate I-52b (1.65 g, 58.50% yield) as a white solid. LCMS: m / z (ESI), [M-Boc] + =243.95.

[0491] Step 2: Add Zn(CN)₂ (417.55 mg, 3.49 mmol, 1 equivalent) and Pd(PPh₃)₄ (402.87 mg, 0.35 mmol, 0.1 equivalent) to a mixture of bromide I-52b (1.2 g, 3.49 mmol, 1 equivalent) in DMF (4.0 mL). Stir the resulting mixture at 70 °C under a nitrogen atmosphere for 2 hours. After cooling to room temperature, dilute the mixture with water (40 mL). Extract the mixture with EtOAc (3 x 40 mL). Wash the combined organic layers with water (3 x 10 mL) and dry over anhydrous Na₂SO₄. Filter and concentrate the filtrate under reduced pressure. The residue was purified by preparative TLC with PE / EA (3:1, v / v) to give methyl 2-[(tert-butoxycarbonyl)amino]-2-(3-cyanophenyl)acetate I-52c (990 mg, 97.81% yield) as a white solid. LCMS: m / z (ESI), [M-Boc+H] + =191.00. 1 HNMR(DMSO-d6,400MHz)δ1.40(9H,s),3.72(3H,s)5.38(1H,d),7.58(1H,t),7.75(1H,m),7.81(1H,d),7.88(1H,d),7.94(1H,d).

[0492] Step 3: Add 1.0 mL of water containing LiOH·H₂O (572.34 mg, 13.64 mmol, 4 equivalents) to a mixture of ester I-52c (990 mg, 3.41 mmol, 1 equivalent) in THF (4.0 mL). Stir the resulting mixture for 6 hours at room temperature under a nitrogen atmosphere. After the reaction, concentrate the resulting mixture under vacuum and dilute with DMF (2 mL). Purify the residue by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water, 10% to 50% gradient, 10 min; detector, UV 254 nm, 25% hold for 25 min. After evaporation of water, [(tert-butoxycarbonyl)amino](3-cyanophenyl)acetic acid I-52 (540 mg, 57.31% yield) as a white solid. LCMS: m / z (ESI), [M+H] + =277.00.

[0493] Synthesis of intermediate I-53

[0494]

[0495] Step 1: At room temperature and under air, t-BuOK (14.16 g, 126.19 mmol, 2.50 equivalents) and Pd(OAc)₂ (1.13 g, 5.03 mmol, 0.10 equivalents) dppf (5.57 g, 10.08 mmol, 0.20 equivalents) were added to a stirred solution of tert-butyl 4-bromobenzoate (20.11 g, 78.21 mmol, 1.55 equivalents) and methyl cyanoacetate (5 g, 50.46 mmol, 1.00 equivalents) in dioxane (40 mL). The resulting mixture was stirred for 3 hours at 70 °C under nitrogen atmosphere. The reaction was quenched by adding water (100 mL) at room temperature. The resulting mixture was extracted with EtOAc (3 x 50 mL). The combined organic layers were dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using PE / EA (1:1, v / v) elution to give tert-butyl 4-(1-cyano-2-methoxy-2-oxoethyl)benzoate I-53b (3.77 g, 27.14% yield), which was a yellow oil. 1 H NMR: (DMSO-d6, 400MHz) δ1.55(9H,s),3.72(3H,d),5.84(1H,s),7.54–7.61(2H,m),7.94–8.02(2H,m).

[0496] Step 2: At 0°C, NaBH4 (4.14 g, 109.55 mmol, 8 equivalents) was added in portions to a stirred solution of nitrile I-53b (3.77 g, 13.69 mmol, 1 equivalent) and CoCl2·6H2O (9.77 g, 41.08 mmol, 3 equivalents) and (Boc)2O (8.97 g, 41.08 mmol, 3 equivalents) in MeOH (50 mL). The resulting mixture was stirred overnight at room temperature. The reaction was quenched by adding ice / water (50 mL) at room temperature. The precipitated solid was filtered off and washed with DCM (3 x 20 mL). The resulting mixture was extracted with CH2Cl2 (3 x 50 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using PE / EA (1:1, v / v) elution to give tert-butyl 4-{3-[(tert-butoxycarbonyl)amino]-1-methoxy-1-oxopropyl-2-yl}benzoate I-53c (2947 mg, 56.71% yield), a yellow oil. LCMS: m / z (ESI), [M+H] + =380.45. 1 H NMR: (CD3OD, 400MHz) δ1.41(9H,s),1.61(9H,s),3.43(1H,dd),3.63(1H,dd),3.70(3H,s),3.98(1H,t),7.40(2H,d),7.89–7.97(2H,m).

[0497] Step 3: At room temperature and under air, LiOH (741.50 mg, 30.96 mmol, 4 equivalents) was added to a stirred solution of ester I-53c (2937 mg, 7.74 mmol, 1 equivalent) in MeOH (20 mL) and H₂O (10 mL). The resulting mixture was stirred for 3 hours at room temperature and under air. The resulting mixture was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water, 90% to 100% gradient, 10 min; detector, UV 254 nm. This yielded 3-[(tert-butoxycarbonyl)amino]-2-[4-(tert-butoxycarbonyl)phenyl]propionic acid I-53 (2.5 g, 88.39% yield) as a white solid. LCMS: m / z (ESI), [M+H] + =366.20. 1H NMR: (CD3OD, 400MHz) δ1.41(9H,d),1.60(8H,s),2.88(1H,s),3.01(1H,s),3.44(1H,dd),3.53(1H,dd),3.73(1H,q),7.46(2H,d),7.88(2H,d).

[0498] Synthesis of intermediate I-54

[0499]

[0500] Step 1: At 0°C and under a nitrogen atmosphere, add Boc₂O (430.26 mg, 1.97 mmol, 3 equivalents) to a solution of methyl 3-[(tert-butoxycarbonyl)amino]-2-(3-cyanophenyl)propionate I-3d (200 mg, 0.66 mmol, 1 equivalent) and CoCl₂·6H₂O (469.05 mg, 1.97 mmol, 3 equivalents) in MeOH (10 mL). At 0°C, add NaBH₄ (124.30 mg, 3.29 mmol, 5 equivalents) to the mixture. Stir the reaction mixture at 0°C for 2 hours. Filter the resulting mixture and wash the filter cake with MeOH (3 x 20 mL). Concentrate the filtrate under reduced pressure. The residue was purified by preparative TLC with PE:EA (2:1, v / v) to give methyl 3-[(tert-butoxycarbonyl)amino]-2-(3-{[(tert-butoxycarbonyl)amino]methyl}phenyl)-propionate I-54a (180 mg, 47.94% yield) as a white solid. LCMS: m / z (ESI), [M+H] + =409.10.

[0501] Step 2: At room temperature and under a nitrogen atmosphere, LiOH·H₂O (36.98 mg, 0.88 mmol, 2 equivalents) was added to a solution of ester I-54a (180 mg, 0.44 mmol, 1 equivalent) in THF (10 mL) and H₂O (5 mL). The mixture was stirred for 2 hours at room temperature. The resulting mixture was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, ACN / water, 10% to 50% gradient, 10 min; detector, UV 220 nm. This yielded 3-[(tert-butoxycarbonyl)amino]-2-(3-{[(tert-butoxycarbonyl)amino]methyl}phenyl)-propionic acid I-54 (159 mg, 91.27% yield) as a white solid. LCMS: m / z (ESI), [M+H] + =395.10.

[0502] Synthesis of intermediate I-55

[0503]

[0504] Step 1: At room temperature and under a nitrogen atmosphere, dppf (2229.81 mg, 4.04 mmol, 0.2 equivalents) and methyl cyanoacetate (2 g, 20.18 mmol, 1 equivalent) were added dropwise to a stirred mixture of 1-(benzyloxy)-4-bromobenzene (8232.15 mg, 31.29 mmol, 1.55 equivalents), t-BuOK (5662.23 mg, 50.46 mmol, 2.5 equivalents), and Pd(OAc)2 (453.15 mg, 2.02 mmol, 0.1 equivalents) in dioxane (50.00 mL, 590.18 mmol) (2.5 equivalents) added dropwise. The resulting mixture was stirred at 70 °C under a nitrogen atmosphere for 3 hours. The resulting mixture was then concentrated under reduced pressure. The residue was purified by silica gel column chromatography using PE / EA (4:1, v / v) elution to give methyl 2-[4-(benzyloxy)phenyl]-2-cyanoacetate I-55b (1.955 g, 34.43% yield) as a yellow solid. 1 H-NMR: (DMSO-d6, 400MHz) δ3.72(3H,s),5.13(2H,s),5.59(1H,s),7.09(2H,m),7.40(7H,m).

[0505] Step 2: At 0°C and under air, Boc₂O (1163.74 mg, 5.33 mmol, 1 equivalent) and NaBH₄ (168.10 mg, 4.44 mmol, 2.5 equivalent) were added in portions to a stirred mixture of nitrile I-55b (500 mg, 1.78 mmol, 1 equivalent) and CoCl₂·6H₂O (1268.63 mg, 5.33 mmol, 3 equivalent) in MeOH (2 mL). The resulting mixture was stirred for 1 hour at room temperature and under air. The reaction was quenched with water at room temperature. The resulting mixture was filtered, and the filter cake was washed with MeOH (2 x 4 mL). The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using PE / EA (3:1, v / v) elution to give methyl 2-[4-(benzyloxy)phenyl]-3-[(tert-butoxycarbonyl)amino]propionate I-55c (371 mg, 54.15% yield) as a white solid. LCMS: m / z (ESI), [M- t Bu] + =330.15.

[0506] Step 3: At room temperature and under air atmosphere, a mixture of ester I-55c (371 mg, 0.96 mmol, 1 equivalent) and LiOH·H₂O (121.16 mg, 2.89 mmol, 3 equivalents) in THF (4 mL) and H₂O (1 mL) was stirred for 2 hours. The resulting mixture was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water, 10% to 50% gradient, 50 min; detector, UV 254 nm. This yielded 2-[4-(benzyloxy)phenyl]-3-[(tert-butoxycarbonyl)amino]propionic acid I-55 (150 mg, 41.96% yield) as a yellow solid. LCMS: m / z (ESI), [MH] - =370.10. 1 H NMR: (DMSO-d6,400MHz)δ1.34(9H,s),3.10–3.24(3H,m),5.05(2H,s),6.50(1H,d),6.82– 6.92(2H,m),7.10–7.17(2H,m),7.28–7.35(1H,m),7.35–7.41(2H,m),7.41–7.46(2H,m).

[0507] Synthesis of intermediate I-56

[0508]

[0509] Step 1: At room temperature, add cyclohexaneacetic acid I-56a (2500 mg, 17.58 mmol, 1.0 equivalent) and SOCl2 (1.5 mL, 20.68 mmol) to a 50 mL round-bottom flask. Stir the resulting mixture at 80 °C under a nitrogen atmosphere for 1 hour. Cool the reaction mixture to room temperature. At room temperature, add phosphorus tribromide (4.26 mL, 44.83 mmol, 2.55 equivalent) and bromine (3512.00 mg, 21.98 mmol, 1.25 equivalent) to the above mixture. Stir the resulting mixture at 80 °C for another 2 hours. Cool the mixture to room temperature. Then, add MeOH (2.5 mL) to the above solution. Stir the resulting mixture at 70 °C under a nitrogen atmosphere for 2 hours. Cool the mixture to room temperature. Quench the reaction with saturated NaS2O3 (aqueous solution) (50 mL) at room temperature. Extract the resulting mixture with DCM (2 x 50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, and evaporated to obtain a crude solid. The crude solid was purified by silica gel column chromatography using PE / EA (5:1, v / v) elution to give methyl 2-bromo-2-cyclohexylacetate I-56b (2000 mg, 40.25% yield) as a grayish-white oil.1 H NMR: (CDCl3, 400MHz) δ0.90–1.21(4H,m),1.27(2H,qt),1.60–1.81(5H,m),1.88(1H,tdt),2.06(1H,dtd),3.77(3H,s),4.02(1H,d).

[0510] Step 2: At room temperature, add 20.0 mL of ACN containing bromide I-56b (2000 mg, 8.52 mmol, 1 equivalent), 1-methylpiperazine (1022.44 mg, 10.21 mmol, 1.2 equivalent), and K₂CO₃ (3526.84 mg, 25.52 mmol, 3.0 equivalent) to a 40 mL vial. Stir the resulting mixture at 80 °C under a nitrogen atmosphere for 12 hours. Cool the mixture to room temperature. Concentrate the resulting mixture under reduced pressure. Purify the residue by silica gel column chromatography using CH₂Cl₂ / MeOH (20:1, v / v) elution to give methyl 2-cyclohexyl-2-(4-methylpiperazin-1-yl)acetate I-56c (950 mg, 24.23% yield) as a grayish-white solid. LCMS: m / z (ESI), [M+H] + =255.15. 1 H NMR: (CDCl3, 400MHz) δ0.83–0.99(3H,m),1.15–1.32(4H,m),1.53(1H,d),1.78(4H,d td),1.95(1H,d),2.30(3H,s),2.53(6H,dd),2.64(2H,dt),2.89(1H,d),3.68(3H,s).

[0511] Step 3: At 70°C under a nitrogen atmosphere, a solution of methylpiperazine I-56c (590 mg, 2.32 mmol, 1 equivalent) and LiOH·H₂O (194.66 mg, 4.64 mmol, 2 equivalents) in MeOH (3.0 mL) and H₂O (3.0 mL) was stirred for 2 hours. The mixture was cooled to room temperature. The mixture was acidified to pH 5 with 1 M HCl (aqueous solution). The resulting mixture was concentrated under vacuum. The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, MeOH / water, 10% to 50% gradient, 30 min; detector, UV 220 nm. This yielded cyclohexyl(4-methylpiperazin-1-yl)acetic acid I-56 (240 mg, 35.8% yield) as a grayish-white solid. LCMS: m / z (ESI), [M+H] + =241.15.

[0512] Synthesis of intermediate I-57

[0513]

[0514] Step 1: At 0°C and under air, NaBH4 (919.81 mg, 24.31 mmol, 1 equivalent) was added in portions to a stirred mixture of methyl 3-[(tert-butoxycarbonyl)amino]-2-(4-cyanophenyl)propionate I-14c (740 mg, 2.43 mmol, 1 equivalent) and CoCl2·6H2O (1156.98 mg, 4.86 mmol, 2 equivalent) in MeOH (10 mL). The resulting mixture was stirred for 5 hours at room temperature under air. After the reaction, the reaction was quenched with water (30 mL) at 0°C. The resulting mixture was filtered, and the filter cake was washed with MeOH (3 x 10 mL). The filtrate was extracted with EtOAc (3 x 20 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to give the crude product. The crude product was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water, 10% to 50% gradient, 10 min; detector, UV 254 nm, 15% hold for 20 min. After solvent evaporation under reduced pressure, methyl 2-[4-(aminomethyl)phenyl]-3-[(tert-butoxycarbonyl)amino]propionate I-57b (300 mg, 40.01% yield) was given as a white solid. LCMS: m / z (ESI), [2M+H] + =617.40.

[0515] Step 2: At 0°C and under air, Et3N (196.89 mg, 1.95 mmol, 3 equivalents) was added in portions to a stirred mixture of amine I-57b (200 mg, 0.649 mmol, 1 equivalent) and acetyl chloride (51.93 mg, 0.66 mmol, 1.02 equivalents) in a DCM (3.0 mL). The resulting mixture was stirred for 3 hours at room temperature under air. After the reaction, the mixture was diluted with water (10 mL). The mixture was extracted with CH2Cl2 (3 x 20 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC using CH₂Cl₂ / MeOH (30:1, v / v) to give methyl 3-[(tert-butoxycarbonyl)amino]-2-[4-(acetamidomethyl)phenyl]propionate I-57c (100 mg, 44.00% yield) as a white solid. LCMS: m / z (ESI), [M-Boc] + =251.15.

[0516] Step 3: At room temperature and under air atmosphere, water (1.0 mL) containing LiOH·H₂O (71.85 mg, 1.71 mmol, 4 equivalents) was added dropwise to a stirred mixture of ester I-57c (150 mg, 0.43 mmol, 1 equivalent) in MeOH (4.0 mL). The resulting mixture was stirred overnight at 60 °C under air atmosphere. After cooling to room temperature, the resulting mixture was concentrated under reduced pressure. The resulting mixture was diluted with MeOH (2.0 mL). The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water, 10% to 50% gradient, 10 min; detector, UV 254 nm, 26% hold for 30 min. After evaporation of the solvent under reduced pressure, 3-[(tert-butoxycarbonyl)amino]-2-[4-(acetamidomethyl)phenyl]propionic acid I-57 (92 mg, 63.89% yield) was given as a white solid. LCMS:m / z(ESI),[M+Na] + =359.00.

[0517] Synthesis of intermediate I-58

[0518]

[0519] Step 1: At room temperature, methyl 3-[(tert-butoxycarbonyl)amino]-2-(4-{[(tert-butyldimethylsilyl)oxy]methyl}phenyl)propionate I-1e (1.5 g, 3.54 mmol, 1 equivalent), TBAF (2.78 g, 10.62 mmol, 3 equivalents), and THF (25 mL) were added to a 40 mL vial. The resulting mixture was stirred for 3 hours at room temperature under air atmosphere. The reaction was quenched by adding water (50 mL) at room temperature. The aqueous layer was extracted with EtOAc (3 x 50 mL). The resulting mixture was concentrated under reduced pressure. The residue was purified by preparative TLC with CH2Cl2 / EA (1:1, v / v) to give methyl 3-[(tert-butoxycarbonyl)amino]-2-[4-(hydroxymethyl)phenyl]propionate I-58a (975 mg, 89.01% yield) as a yellow oil. LCMS:m / z(ESI),[M+H] + =310.20. 1 H NMR (CD3OD, 400MHz) δ1.42(9H,s),3.39(1H,m),3.64(5H,m),4.60(2H,s),5.51(1H,s),7.31(4H,m).

[0520] Step 2: At room temperature, methyl 3-[(tert-butoxycarbonyl)amino]-2-[4-(hydroxymethyl)phenyl]propionate (650 mg, 2.10 mmol, 1 equivalent), N-methylcarbamoyl chloride (432.24 mg, 4.62 mmol, 2.2 equivalents), TEA (1.06 g, 10.51 mmol, 5 equivalents), and DCM (15 mL) were added to a 40 mL vial. The resulting mixture was stirred for 3 hours at room temperature under air. The mixture was then concentrated under reduced pressure. The residue was purified by preparative TLC with CH2Cl2 / MeOH (40:1, v / v) to give methyl 3-[(tert-butoxycarbonyl)amino]-2-(4-{[(methylcarbamoyl)oxy]methyl}phenyl)-propionate I-58b (570 mg, 74.04% yield) as a yellow oil. LCMS:m / z(ESI),[M+H] + =367.05.

[0521] Step 3: At room temperature, ester I-58b (570 mg, 1.56 mmol, 1.00 equivalent), LiOH·H₂O (163.18 mg, 3.89 mmol, 2.5 equivalent), MeOH (4 mL), and water (2 mL) were added to a 100 mL round-bottom flask. The mixture was stirred for 16 hours at room temperature under air. The mixture was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, MeOH / water (0.1% FA), 0% to 25% gradient, 20 min; detector, UV 254 nm. The mixture was concentrated under reduced pressure to give 3-[(tert-butoxycarbonyl)amino]-2-(4-{[(methylcarbamoyl)-oxy]methyl}phenyl)propionic acid I-58 (520 mg, 94.86% yield) as a grayish-white solid. LCMS:m / z(ESI),[M+H-Boc] + =253.15.

[0522] Synthesis of intermediate I-59

[0523]

[0524] Step 1: At 0°C, add 10 mL of DMF containing methyl 2-(3-methoxyphenyl)acetate I-59a (3 g, 16.65 mmol, 1 equivalent), t-BuOK (3.74 g, 33.30 mmol, 2 equivalents), and tert-butyl 4-iodopiperidine-1-carboxylate (6.73 g, 21.642 mmol, 1.3 equivalents) to a 50 mL round-bottom flask. Stir the resulting mixture at room temperature for 2 hours. Purify the residue by silica gel column chromatography using PE / EA (25:1, v / v) elution to give piperidine I-59b (2.02 g, 33.38%) as a yellow oil. LCMS: m / z (ESI), [M+H- t Bu] + =308.20. 1 H NMR(DMSO-d6,400MHz)δ0.83–0.97(1H,m),1.04–1.17(2H,m),1.37(9H,s),1.65(1H,dt),2.07(1H,qt),2. 66(2H,d),3.38(1H,d),3.59(3H,s),3.83(1H,d),3.90–3.98(1H,m),6.81–6.92(3H,m),7.19–7.30(1H,m).

[0525] Step 2: At room temperature, add 3 mL of DCM containing piperidine I-59b (2.0 g, 5.50 mmol, 1 equivalent) and TFA (1.5 mL) to a 50 mL round-bottom flask. Stir the resulting mixture at room temperature for 1.5 hours. Alkalinize the mixture to pH 9 with saturated NaHCO3 (aqueous solution). Extract the resulting mixture with DCM (3 x 50 mL). Concentrate the combined organic layers under reduced pressure to give piperidine I-59c (1.3 g, 89.71%) as a yellow oil. LCMS: m / z (ESI), [M+H] + =264.20. 1 H NMR(DMSO-d6,400MHz)δ1.08–1.23(1H,m),1.23–1.33(1H,m),1.33–1.46(1H,m),1.81(1H,dt),2.22(1H,qt),2.77(1H,td),2 .87(1H,td),3.15(1H,dt),3.26(1H,dt),3.43(1H,d),3.57(3H,s),3.75(3H,s),6.81–6.95(3H,m),7.28(1H,t),8.25(2H,s).

[0526] Step 3: At room temperature, add 2.5 mL of MeOH containing piperidine I-59c (1.3 g, 4.94 mmol, 1 equivalent), DIEA (3.19 g, 24.69 mmol, 5.0 equivalent), sodium deuterated cyanoboroborate (620 mg, 9.87 mmol, 2 equivalent), and CH2O (296 mg, 9.88 mmol, 2 equivalent) to a 50 mL round-bottom flask. Stir the resulting mixture overnight at room temperature. Quench the reaction with water at room temperature. Extract the resulting mixture with EtOAc (3 x 10 mL). Concentrate the combined organic layers under reduced pressure to give methylpiperidine I-59d (1.1 g, 80.34%) as a yellow oil. LCMS: m / z (ESI), [M+H] + =278.25.

[0527] Step 4: At room temperature, add 2 mL of THF containing methylpiperidine I-59d (600 mg, 2.16 mmol, 1 equivalent) and 2 mL of water containing LiOH (104 mg, 4.33 mmol, 2 equivalents) to a 20 mL vial. Stir the resulting mixture at room temperature for 3 hours. Acidify the mixture to pH 5 with HCl (1 M). Purify the residue by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, water / MeOH, 0% to 30% gradient, 40 min; detector, UV 254 nm, to obtain acid I-59 (270 mg, 47.40%) as a white solid. LCMS: m / z (ESI), [M+H) + =264.20. 1 H NMR(DMSO-d6,400MHz)δ0.95–1.16(2H,m),1.27(1H,qd),1.63–1.97(4H,m),2.14(3H,s ),2.67(1H,d),2.79(1H,d),3.14(1H,d),3.73(3H,s),6.71–6.94(3H,m),7.23(1H,t).

[0528] Synthesis of intermediate I-60

[0529]

[0530] Step 1: At room temperature, ester I-60a (2 g, 11.10 mmol, 1 equivalent), formaldehyde (0.67 g, 22.20 mmol, 2 equivalents), K₂CO₃ (4.60 g, 33.30 mmol, 3 equivalents), and DMF (2 mL) were added to a 20 mL vial. The mixture was stirred at 85 °C for 1 hour. The mixture was cooled to room temperature. The resulting mixture was diluted with EtOAc (30 mL) and washed with water (3 x 30 mL). The combined organic layers were dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC with PE / EA (8:1, v / v) to give acrylate I-60b (1.3 g, 60.88%) as a pale yellow oil. 1 H NMR (CD3OD, 400MHz) δ3.82(6H,d),5.95(1H,d),6.31(1H,d),6.88–6.95(1H,m),6.95–7.02(2H,m),7.23–7.31(1H,m).

[0531] Step 2: At room temperature, add enolate I-60b (1.3 g, 6.76 mmol, 1 equivalent), 1-methylpiperazine (2.71 g, 27.05 mmol, 4 equivalents), TEA (2.05 g, 20.29 mmol, 3 equivalents), and DMF (10 mL) to a 100 mL round-bottom flask. Stir the resulting mixture at room temperature for 1 hour. Dilute the resulting mixture with EtOAc (50 mL) and wash with 2 x 30 mL water and 1 x 30 mL saturated brine. Concentrate the resulting mixture under reduced pressure. This yields methylpiperidine I-60c (1.7 g, 81.33%) as a brownish-yellow oil. LCMS: m / z (ESI), [M+H ]+ =293.15.

[0532] Step 3. At room temperature, add methylpiperidine I-60c (1 g, 3.42 mmol, 1 equivalent), LiOH (163.83 mg, 6.84 mmol, 2 equivalents), THF (5 mL), and water (5 mL) to an 80 mL vial. Stir the resulting mixture at room temperature for 1 hour. Acidify the mixture to pH 5 with HCl (1 M). Purify the residue by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, MeOH / water, 10% to 50% gradient, 30 min; detector, UV 220 nm. This yields acid I-60 (497 mg, 51.16%) as a white solid. LCMS: m / z (ESI), [M+H) + =279.15.

[0533] Compound Synthesis

[0534] Example 1 (racemate), Example 2 (enantiomer 1), and Example 3 (enantiomer 2)

[0535] Preparation of N-(3-(1H-pyrazol-4-yl)-1H-indol-7-yl)-3-amino-2-(4-(hydroxymethyl)phenyl)propionamide

[0536]

[0537] Step 1: At room temperature and under a nitrogen atmosphere, a mixture of acid I-1 (498.91 mg, 1.69 mmol, 1.2 equivalents), amine I-49 (420 mg, 1.41 mmol, 1 equivalent), HATU (802.92 mg, 2.11 mmol, 1.5 equivalents), and triethylamine (427.37 mg, 4.22 mmol, 3 equivalents) in DMF (5 mL) was stirred overnight. The resulting mixture was diluted with ethyl acetate (100 mL). The residue was washed with saturated NaHCO3 (1 x 80 mL), water (2 x 80 mL), and saturated NaCl (1 x 80 mL). The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water, 35% to 65% gradient, 40 min; detector, UV 254 nm, to give amide C-1a (500 mg, 61.70%) as a grayish-white solid. LCMS:m / z(ESI),[M+H] + =576.5. 1 H NMR(DMSO-d6,400MHz)δ1.37(9H,s),1.62(9H,s),3.60(1H,s),4.08(1H,s),4.47(2H,s),5.13(1H,s),7.04(1H,t),7.10(1H ,s),7.29(2H,d),7.38(2H,d),7.45(1H,d),7.60(1H,d),7.84(1H,d),8.26(1H,s),8.49(1H,s),9.97(1H,s),10.83(1H,s).

[0538] Step 2: Amide C-1a (250 mg) was purified by preparative HPLC under the following conditions (column: CHIRALPAKID, 2*25 cm, 5 μm; mobile phase A: Hex (0.2% DEA), mobile phase B: EtOH; flow rate: 20 mL / min; gradient: 50% B, 13 min; wavelength: 220 / 254 nm; sample solvent: EtOH) to obtain the enantiomer amide C-1b (80 mg, 32.00%) as a grayish-white solid. R= 7.48 minutes. LCMS:m / z(ESI),[M+H] + =576.2. 1 H NMR(CD3OD,400MHz)δ1.44(9H,s),1.70(9H,s),3.48(1H,dd),3.79(1H,dd),4.11(1H,dd),4.62( 2H,s),7.05–7.14(2H,m),7.38(2H,d),7.49(2H,d),7.64(2H,q),8.16(1H,d),8.42(1H,d), SFC:t R =1.41 minutes, ee=100%, and the enantiomer C-1c (85.3 mg, 32.52%) as a grayish-white solid. t R = 9.78 minutes. LCMS:m / z(ESI),[M+H] + =576.2. 1 H NMR(CD3OD,400MHz)δ1.44(9H,s),1.70(9H,s),3.48(1H,dd),3.79(1H,dd),4.11(1H,dd),4.6 2(2H,s),7.05–7.14(2H,m),7.38(2H,d),7.49(2H,d),7.64(2H,q),8.16(1H,d),8.42(1H,d). SFC:t R =1.89 minutes, ee=99.76%.

[0539] Step 3: (Example 3, Enantiomer 2): A solution of enantiomer C-1c (33 mg, 57.33 μmol) in DCM (1.48 mL) was cooled and stirred in an ice / water bath at 0 °C. Trifluoroacetic acid (261.45 mg, 2.29 mmol, 176.66 μL) was then added dropwise to the reaction mixture. The resulting mixture was stirred for 0.5 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by passing it through a C18 rapid column and eluting with a 0% to 60% gradient of MeCN / water (6 mmol / L NH4HCO3). The purified fraction was lyophilized to give enantiomer 2 of amine Example 3 (20.76 mg, 93% yield) as a white solid. LCMS:t R =1.09 min, 3 min chromatography (5-95% MeCN / water (0.02% TFA), waters Acquity UPLC BEH C18 1.7um, 2.1*50mm, 40℃), MS (ESI) m / z = 376.2 [M+H] + . 1H NMR: (400MHz, DMSO-d6) δ12.80(s,1H),10.83(s,1H),7.94(s,2H),7.57(d,J=12.1Hz,2H),7.49–7.31(m,3H),7.29 (d,J=7.9Hz,2H),6.98(t,J=7.8Hz,1H),5.14(s,1H),4.46(s,2H),3.92–3.84(m,1H),2.89(dd,J=11.7,5.0Hz,1H).

[0540] Following the above procedure, starting with amide C1-b, we can obtain Example 2 (enantiomer 1).

[0541] Step 4: (Example 1, racemic mixture): At 0°C, add 3 mL of DCM containing amide C-1a (60 mg, 0.10 mmol, 1 equivalent) and 1 mL of TFA to a 100 mL round-bottom flask. Stir the resulting mixture at 0°C under a nitrogen atmosphere for 1 hour. Concentrate the resulting mixture under reduced pressure. Dissolve the residue in methanol (3 mL). Alkalize the mixture to pH 9 with NH3 (aqueous solution). Purify the crude product by preparative HPLC under the following conditions: (Column: Xselect CSH C18 OBD 30*150 mm; Mobile phase A: water (0.1% FA), Mobile phase B: ACN; Flow rate: 60 mL / min; Gradient: 5% B to 18% B, 7 min; Wavelength: 254 nm; 220 nm; t R =6.05), to obtain Example 1 (21.5 mg, 54.94%), a racemic mixture that was a white solid. LCMS: m / z (ESI), [M+H) + =376.3. 1 H NMR (CD3OD, 400MHz) δ3.37(1H,s),3.68(1H,dd),4.26(1H,dd),4.65(2H,s),7.04–7.12(2H,m),7.43–7.55(5H,m),7.62–7.69(1H,m),7.92(2H,s).

[0542] Example 5 (enantiomer 1) and Example 6 (enantiomer 2)

[0543] Preparation of 4-(1-((3-(1H-pyrazol-4-yl)-1H-indol-7-yl)amino)-3-amino-1-oxopropyl-2-yl)benzyl ester of 3,5-dimethylbenzoic acid

[0544]

[0545] Step 1: At room temperature, add 5 mL of DMF containing alcohol C-1a (146 mg, 0.25 mmol, 1 equivalent) and 2,4-dimethylbenzoic acid (45.71 mg, 0.31 mmol, 1.2 equivalent), EDCI (72.93 mg, 0.38 mmol, 1.5 equivalent), and DMAP (30.98 mg, 0.25 mmol, 1 equivalent) to a 50 mL round-bottom flask. Stir the resulting mixture for 4 hours at room temperature under a nitrogen atmosphere. Precipitate the product by adding water. Purify the crude product (146 mg) by preparative HPLC under the following conditions: (Column: XBridge Prep OBD C18 30*150 mm, 5 μm; Mobile phase A: water (10 mmol / L NH4HCO3), Mobile phase B: ACN; Flow rate: 60 mL / min; Gradient: 55% B to 85% B, 7 min; Wavelength: 220 nm; t R =8.25 minutes), to obtain a racemic ester C-5a (130 mg, 72.42%) as a grayish-white solid. LCMS: m / z (ESI), [M+H) + =708.30. 1 H NMR(DMSO-d6,400MHz)δ1.36(9H,s),1.62(9H,s),2.31(3H,s),2.50(3H,s),3.35–3.42(1H,m),3.61(1H,dt),4.12(1H,t),5.28(2H,s ),6.97–7.16(4H,m),7.40–7.53(5H,m),7.61(1H,d),7.78(1H,d),7.84(1H,d),8.27(1H,s),8.49(1H,s),10.02(1H,s),10.83(1H,s).

[0546] Step 2: Purification of the racemic ester C-5a (260 mg) by preparative chiral HPLC under the following conditions (column: (R,R)WHELK-O1, 4.6*50 mm, 3.5 μm; mobile phase A: Hex (0.2% DEA): (EtOH / DCM = 1:1, v / v) = 65:35 (v / v); flow rate: 1 mL / min; injection volume: 5 μL / mL) to obtain the enantiomer ester C-5c, SFC:t R =2.79 minutes, ee=100%, and enantiomer ester C-5b, SFC:t R =3.567 minutes, ee=100%.

[0547] Step 3: At room temperature, add 3 mL of DCM containing enantiomer ester C-5c (110 mg, 0.16 mmol, 1 equivalent) and 1 mL of TFA to a 100 mL vial. Stir the resulting mixture for 30 minutes at room temperature under a nitrogen atmosphere. Concentrate the resulting mixture under reduced pressure. Dissolve the residue in methanol (3 mL). Alkalize the residue to pH 8 with an aqueous NH3 solution. Stir the resulting mixture for 1 hour at 0 °C under a nitrogen atmosphere. Purify the crude product (80 mg) by preparative HPLC under the following conditions: (Column: XBridge Shield RP18 OBD 30*150 mm, 5 μm; Mobile phase A: water (10 mmol / L NH4HCO3), Mobile phase B: ACN; Flow rate: 60 mL / min; Gradient: 30% B to 60% B, 8 min; Wavelength: 220 nm; t R =7.58 minutes), to obtain enantiomer 2 of Example 6 (28.8 mg, 36.51%) as a white solid. LCMS: m / z (ESI), [M+H) + =508.25. 1 H NMR(CD3OD,400MHz)δ2.35(3H,s),2.54(3H,s),3.08(1H,dd),3.46(1H,dd),4.01(1H,dd),5.33(2H,s),6.9 8–7.08(2H,m),7.08–7.15(2H,m),7.45(1H,s),7.48–7.57(4H,m),7.64(1H,dd),7.83(1H,d),7.91(2H,s). SFC:t R =2.338 minutes, ee = 99.28%.

[0548] At room temperature, add 3 mL of DCM and 1 mL of TFA containing the enantiomer ester C-5b (110 mg, 0.16 mmol, 1 equivalent) to a 100 mL vial. Stir the resulting mixture for 30 min at room temperature under a nitrogen atmosphere. Concentrate the mixture under reduced pressure. Dissolve the residue in methanol (3 mL). Alkalize the residue to pH 8 with an aqueous NH3 solution. Stir the resulting mixture for 1 h at 0 °C under a nitrogen atmosphere. Purify the crude product by preparative HPLC under the following conditions: (Column: XBridge Prep OBD C18 30*150 mm, 5 μm; Mobile phase A: water (10 mmol / L NH4HCO3), Mobile phase B: ACN; Flow rate: 60 mL / min; Gradient: 34% B to 54% B, 8 min; Wavelength: 254 nm; t R=7.83 minutes), to obtain enantiomer 1 of Example 5 (24.5 mg, 42.71%) as a white solid. LCMS: m / z (ESI), [M+H) + =508.25. 1 H NMR(CD3OD,400MHz)δ2.35(3H,s),2.55(3H,s),3.04(1H,dd),3.43(1H,dd),3.96(1H,d d),5.34(2H,s),7.03–7.15(4H,m),7.42–7.54(5H,m),7.61–7.85(2H,m),7.91(2H,s). SFC:t R =2.123 minutes, ee=100%.

[0549] Example 7 (racemic mixture)

[0550] Preparation of N-(3-(1H-pyrazol-4-yl)-1H-indol-7-yl)-2-amino-2-phenylacetamide (racemate)

[0551]

[0552] Step 1: Iron powder (3.16 g, 56.52 mmol, 401.63 μL) was added to a mixture of nitro-indole I-7a (5 g, 11.30 mmol) and ammonium hydrochloride (6.05 g, 113.05 mmol) in EtOH (30 mL) and water (30 mL). The reaction mixture was then stirred and heated at 80 °C for 3 hours. The reaction mixture was filtered through diatomaceous earth and washed with ethanol (50 mL x 3), and the filtrate was concentrated. The residue was dissolved in 200 mL of ethyl acetate, filtered, and the filtrate was concentrated under reduced pressure to give amine I-7b (3.10 g, 67% yield) as a brown solid. LCMS:t R =2.07 min, 3 min chromatography (3 min - 5 - 95% MeCN / water (0.02% TFA), Waters Acquity UPLC BEH C18 1.7um, 2.1*50mm, 40℃), MS (ESI) m / z = 413.2 [M+H] + .

[0553] Step 2: At room temperature, NMP (20 mL) containing amine I-7b (3.1 g, 7.52 mmol), 2-(tert-butoxycarbonylamino)-2-phenyl-acetic acid (3.21 g, 12.78 mmol), and HATU (4.86 g, 12.78 mmol) was stirred for 15 minutes. Then, DIPEA (4.37 g, 33.83 mmol, 5.89 mL) was added at room temperature. The resulting suspension was stirred for 16 hours at room temperature. The reaction mixture was diluted with 200 mL of water, stirred for 30 minutes at room temperature, filtered, and washed with water (10 mL x 2). The solid was collected and dried to give a brown solid racemic amide C-7a (1.60 g, 33% yield). LCMS:t R =2.13 min, 3 min chromatography (3 min - 5 - 95% MeCN / water (0.02% TFA), Waters Acquity UPLC BEH C18 1.7um, 2.1*50mm, 40℃), MS (ESI) m / z = 646.7 [M+H] + .

[0554] Step 3: At 90°C and under nitrogen, dioxane (3.00 mL) containing racemic C-7a (200 mg, 309.80 μmol), 4-bromo-1-toluenesulfonyl-1H-pyrazole (139.95 mg, 464.70 μmol), K₂CO₃ (128.45 mg, 929.41 μmol), and PdCl₂dppf (25.30 mg, 30.98 μmol) and water (0.60 mL) was stirred for 16 hours. The solvent was removed under reduced pressure. The residue was purified by passing it through a silica gel column and eluting with a gradient of ethyl acetate / petroleum ether from 0% to 40%. The purified fraction was evaporated to dryness to give racemic amide C-7b (136 mg, 59% yield) as a brown solid. LCMS:t R =2.01 min, 3 min chromatography (3 min - 5 - 95% MeCN / water (0.02% TFA), Waters Acquity UPLC BEH C18 1.7um, 2.1*50mm, 40℃), MS (ESI) m / z = 740.7 [M+H] + .

[0555] Steps 4 and 5: Racemic amide C-7b (136 mg, 183.82 μmol) was inserted into THF (0.60 mL) / MeOH (0.60 mL), followed by the addition of water containing 2N sodium hydroxide (73.53 mg, 1.84 mmol, 34.52 μL). The reaction mixture was stirred for 3 hours at room temperature. When the reaction was complete, the mixture was stirred and cooled to 0°C in an ice / water bath. Then, saturated NH4Cl aqueous solution (20 mL) was added, and the mixture was extracted with dichloromethane (10 mL x 3). The combined organic layers were washed with brine (50 mL x 1), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain crude racemic amide C-7c. The crude racemic amide was dissolved in DCM (2.00 mL). The reaction mixture was stirred and cooled to 0°C in an ice / water bath. Then, trifluoroacetic acid (1.05 g, 9.19 mmol, 708.08 μL) was added dropwise to the reactants. The resulting mixture was stirred at 0 °C for 2 hours. The mixture was then concentrated under reduced pressure. The residue was purified by C18 rapid chromatography with a 0% to 30% gradient elution of MeCN / water (0.02% TFA). The purified fractions were lyophilized to give the racemic mixture Example 7 (3.5 mg, 7.88 μmol, 4.28% yield, TFA salt) as a brown solid. LCMS:t R =0.93 min, 3 min chromatography (3 min - 5 - 95% MeCN / water (0.02% TFA), Waters Acquity UPLC BEH C18 1.7um, 2.1*50mm, 40℃), MS (ESI) m / z = 332.4 [M+H] + . 1 H NMR: (400MHz, DMSO-d6) δ8.42(s,2H),8.36(d,J=7.7Hz,1H),8.29–8.25(m,1H),8.16(dd,J=7.5,1.9Hz,1H) ,8.10(d,J=7.8Hz,1H),7.96–7.84(m,5H),7.60(dd,J=7.4,1.0Hz,1H),7.46(t,J=7.4Hz,1H),6.35(s,1H).

[0556] Example 8

[0557] Preparation of (2R)-2-amino-2-phenyl-N-[3-(1H-pyrazol-4-yl)-1H-indole-7-yl]acetamide

[0558]

[0559] Step 1: At room temperature, amine I-49 (120 mg, 0.40 mmol, 1 equivalent), (R)-[(tert-butoxycarbonyl)amino](phenyl)acetic acid (151.61 mg, 0.60 mmol, 1.5 equivalent), TCFH (225.71 mg, 0.80 mmol, 2 equivalent), NMI (165.12 mg, 2.01 mmol, 5 equivalent), and MeCN (5 mL) were added to an 80 mL vial. The resulting mixture was stirred at room temperature for 8 hours. The mixture was then concentrated under reduced pressure. The residue was purified by preparative TLC with CH2Cl2 / methanol (40:1, v / v) to give amide (R)-C-8 (160 mg, 83.39%) as a yellow solid. LCMS: m / z (ESI), [M+H- t Bu] + =476.20.

[0560] Step 2: At room temperature, add amide(R)-C-8 (120 mg, 0.23 mmol, 1 equivalent), TFA (1 mL), and DCM (3 mL) to a 50 mL round-bottom flask. Stir the resulting mixture for 1 hour at room temperature. Dilute the mixture with CH2Cl2 (20 mL). Cool the mixture to 0 °C. Alkalize the mixture to pH 9 with an aqueous NH3 solution. Purify the crude product (100 mg) by preparative HPLC under the following conditions: (Column: XBridge Shield RP18 OBD 30*150 mm, 5 μm; Mobile phase A: Water (10 mmol / L NH4HCO3 + 0.1% NH3 aqueous solution), Mobile phase B: ACN; Flow rate: 60 mL / min; Gradient: 17% B to 40% B, 8 min; Wavelength: 220 nm; t R = 7.67 min). The crude product (60 mg) was purified by preparative chiral HPLC under the following conditions: column: CHIRAL ART Cellulose-SB, 2*25 cm, 5 μm; mobile phase A: Hex (0.2% DEA), mobile phase B: EtOH / DCM (1:1, v / v); flow rate: 20 mL / min; gradient: 45% B to 45% B, 16 min; wavelength: 220 / 254 nm; t R -1 = 8.97 minutes; t R-2 = 12.51 min; Sample solvent: EtOH / DCM (1:1, v / v); Injection volume: 0.7 mL). The crude product (32 mg) was purified by preparative HPLC under the following conditions (column: XBridge Shield RP18OBD 30*150 mm, 5 μm; mobile phase A: water (10 mmol / L NH4HCO3 + 0.1% NH3 aqueous solution), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 17% B to 40% B, 8 min, 40% B; wavelength: 220 nm; t R =7.67 minutes), to obtain (R)-Example 8 (17.5 mg, 23.34%) as a white solid. LCMS: m / z (ESI), [M+H] + =332.10. 1 H NMR (CD3OD, 400MHz) δ4.76(1H,s),7.07(1H,t),7.14(1H,dd),7.31–7.39(1H,m),7.39–7.48(3H,m),7.59(2H,dd),7.65(1H,dd),7.92(2H,s). SFC:t R =0.938 minutes, ee=100%.

[0561] Example 9

[0562] Preparation of (2S)-2-amino-2-phenyl-N-[3-(1H-pyrazol-4-yl)-1H-indole-7-yl]acetamide

[0563]

[0564] Step 1: At room temperature and under a nitrogen atmosphere, a solution of amine I-49 (150 mg, 0.50 mmol, 1 equivalent) and (S)-[(tert-butoxycarbonyl)amino](phenyl)acetic acid (189.51 mg, 0.75 mmol, 1.5 equivalent), TCFH (211.60 mg, 0.75 mmol, 1.5 equivalent), and NMI (144.48 mg, 1.76 mmol, 3.5 equivalent) in MeCN (3 mL) was stirred for 1 hour. The residue was purified by preparative TLC with CH2Cl2 / methanol (22:1, v / v) to give amide (S)-C-9 (200 mg, 74.83%) as a yellow solid. LCMS: m / z (ESI), [M+H- t Bu] + =476.25.

[0565] Step 2: At room temperature, under a nitrogen atmosphere, a solution of amide (S)-C-9 (160 mg, 0.27 mmol, 1 equivalent) and TFA (1.00 mL) in DCM (3 mL) was stirred for 1 hour. The resulting mixture was concentrated under reduced pressure. The mixture was diluted with methanol (2 mL). At 0 °C, the mixture was alkalized to pH 9 with ammonia and stirred for 30 minutes. The crude product (200 mg) was purified by preparative HPLC under the following conditions (column: XBridge Shield RP18 OBD 30*150 mm, 5 μm; mobile phase A: water (10 mmol / L NH4HCO3 + 0.1% NH3 aqueous solution), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 17% B to 40% B, 8 min; wavelength: 220 nm; t R =7.77 minutes), to obtain (S)-Example 9 (46.6 mg, 20.74%) as a grayish-white solid. LCMS: m / z (ESI), [M+H] + =332.15. 1 HNMR (CD3OD, 400MHz) δ4.75(1H,s),7.07(1H,t),7.14(1H,d),7.31–7.39(1H,m),7.43(3H,dd),7.59(2H,d),7.65(1H,d),7.91(2H,s). SFC:t R = 1.364 minutes, ee = 100%.

[0566] Example 10 (Enantiomer 1)

[0567] Preparation of methyl 6-nitrooxyhexanoic acid [4-[(1S)-1-(aminomethyl)-2-oxo-2-[[3-(1H-pyrazol-4-yl)-1H-indole-7-yl]amino]ethyl]phenyl]

[0568]

[0569] Step 1: MeCN (12.5 mL) containing 6-bromohexanoic acid C-10a (500 mg, 2.56 mmol) and silver nitrate (566.09 mg, 3.33 mmol, 130.08 μL) was added at 70 °C for 18 hours. The reaction solution was then evaporated under reduced pressure. 10 mL of DCM was then added. The suspension was stirred for 15 minutes at room temperature, filtered, and evaporated under reduced pressure to give 6-nitrooxyhexanoic acid C-10b (440 mg, 97% yield) as a brown oil. 1H NMR: (400MHz, CDCl3) δ11.18(s,1H),4.45(t,J=6.6Hz,2H),2.38(t,J=7.3Hz,2H),1.83–1.63(m,4H),1.56–1.40(m,2H).

[0570] Step 2: 6-Nitrooxyhexanoic acid C-10b (23.08 mg, 130.29 μmol) was dissolved in DCM (1.50 mL) at 0 °C using an ice / water bath. EDCI (24.98 mg, 130.29 μmol), DMAP (1.06 mg, 8.69 μmol), and the enantiomer alcohol C-1c (50 mg, 86.86 μmol) were added. The reaction mixture was stirred and slowly heated to 25 °C for 16 hours. The resulting mixture was diluted with water (50 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic layers were washed with brine (60 mL x 1), dried over anhydrous sodium sulfate, filtered, and evaporated under reduced pressure to give the enantiomer ester C-10c (62 mg, 97% yield) as a brown solid. LCMS:t R =2.07 min, 3 min chromatography (5-95% MeCN / water (6 mmol / L NH4HCO3), waters AcquityUPLC BEH C18 1.7 μm, 2.1*50 mm, 40 °C), MS (ESI) m / z = 735.2 [M+H] + .

[0571] Step 3: At 0 °C, a solution of enantiomer C-10c (62.00 mg, 84.38 μmol) in DCM (3.01 mL) was cooled and stirred in an ice / water bath. Then, TFA (481.05 mg, 4.22 mmol, 325.03 μL) was added dropwise to the reaction mixture. The resulting mixture was stirred for 2 hours. The reaction mixture was concentrated under reduced pressure. The residue was purified by C18 rapid chromatography with a 0% to 50% gradient elution of MeCN / water (6 mmol / L NH4HCO3). The purified fraction was lyophilized to give enantiomer 1 of nitrooxy ester Example 10 (40 mg, 89% yield) as a white solid. LCMS:t R =1.63 min, 3 min chromatography (5-95% MeCN / water (6 mmol / L NH4HCO3), Waters Acquity UPLC BEH C18 1.7 μm, 2.1*50 mm, 40 °C), MS (ESI) m / z = 535.3 [M+H] + . 1H NMR: (500MHz, DMSO-d6) δ12.85(s,1H),10.81(s,1H),7.95(s,2H),7.57(d,J=8.8Hz,2H),7.51–7.20(m,5H),6.98(t,J=7.5Hz,1H),5.06(s,2H ),4.48(t,J=6.2Hz,2H),3.98–3.84(m,1H),3.29–3.26(m,1H),2.98–2. 84(m,1H),2.35(t,J=7.0Hz,2H),1.75–1.47(m,4H),1.42–1.27(m,2H).

[0572] Example 11 (Enantiomer 1)

[0573] Preparation of 4-(1-((3-(1H-pyrazol-4-yl)-1H-indole-7-yl)amino)-3-amino-1-oxopropyl-2-yl)benzyl ester of 5-(nitrooxy)valerate

[0574]

[0575] Step 1: At 70°C and under a nitrogen atmosphere, a mixture of 5-bromopentanoic acid I-11a (1 g, 5.52 mmol, 1 equivalent) and AgNO3 (1.9 g, 11.05 mmol, 2 equivalents) in MeCN (15 mL) was stirred for 24 hours. The reaction is light-sensitive and should be avoided from light. The mixture was cooled to room temperature. The precipitate was collected by filtration and washed with dichloromethane (5 x 40 mL). After filtration, the filtrate was concentrated under reduced pressure to give 600 mg of 5-(nitrooxy)pentanoic acid I-11b (67%) as a colorless oil. 1 H NMR (DMSO-d6, 400MHz) δ1.58-1.63(2H,m),1.79-1.81(2H,m),2.23-2.29(2H,m),3.49-3.54(2H,m),12.08(1H,s).

[0576] Step 2: At 0°C and under a nitrogen atmosphere, EDCI (57 mg, 0.30 mmol, 2 equivalents) and DMAP (2 mg, 0.016 mmol, 0.1 equivalents) were added to a mixture of 5-(nitrooxy)valerate I-11b (120 mg, 0.74 mmol, 5 equivalents) in DCM (10 mL) over 30 minutes, followed by dropwise addition of alcohol C-1c (85 mg, 0.15 mmol, 1 equivalent) at 0°C. The mixture was warmed to room temperature and the reaction mixture was stirred for 16 hours. The reaction mixture was quenched with water (20 mL) at room temperature and extracted with CH2Cl2 (3 x 20 mL). The combined organic layers were washed with saturated NaCl (3 x 20 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC using dichloromethane / methanol (30:1, v / v) elution to give 60 mg of the enantiomer C-11 (40%) as an orange solid. LCMS: m / z (ESI), [M+H] + =721.31.

[0577] Step 3: At 0°C and under a nitrogen atmosphere, the mixture of enantiomer C-11 (60 mg, 0.08 mmol, 1 equivalent) and TFA (0.8 mL, 10.74 mmol) in DCM (3 mL) was stirred for 30 minutes. The resulting mixture was concentrated under reduced pressure. The residue was neutralized to pH 7–8 with saturated NaHCO3 and extracted with CH2Cl2 (3 x 10 mL). The combined organic layers were washed with saturated NaCl (3 x 10 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative HPLC under the following conditions (mobile phase consisting of water containing 0.1% NH4HCO3 and acetonitrile containing 0.1% NH4OH; constant gradient, mobile phase: 70% aqueous / 30% organic to 47% aqueous / 53% organic, 8 min; flow rate constant at 60 mL / min) to obtain 10.6 mg of enantiomer 1 (24%) of Example 11 as an orange solid. LCMS: m / z (ESI), [M+H) + =521.25. 1H NMR(DMSO-d6,400MHz)δ1.60-1.69(4H,m),2.39(2H,t),2.88-2.93(1H,m),3.85-3.92(1H,m),4.50(2H,t),5.07(2H,s),6.96( 1H,t),7.32-7.38(3H,m),7.43(2H,d),7.56-7.60(2H,m),7.96(2H,d),7.82-8.12(2H,m),10.78(1H,s),12.81-12.88(1H,m).

[0578] Example 12 (Enantiomer 1)

[0579] Preparation of 4-(nitrooxy)butyric acid 4-(1-((3-(1H-pyrazol-4-yl)-1H-indole-7-yl)amino)-3-amino-1-oxopropyl-2-yl)benzyl ester

[0580]

[0581] Step 1: At 0°C and under a nitrogen atmosphere, EDCI (133 mg, 0.70 mmol, 2 equivalents) and DMAP (4.2 mg, 0.04 mmol, 0.1 equivalents) were added to a mixture of 4-bromobutyric acid C-12a (290 mg, 1.75 mmol, 5 equivalents) in DCM (30 mL) for 30 minutes, followed by the addition of alcohol C-1c (200 mg, 0.35 mmol, 1 equivalent). The reaction mixture was stirred for 2 hours at room temperature. The mixture was quenched with water (30 mL), extracted with CH2Cl2 (3 x 30 mL), washed with saturated NaCl (3 x 20 mL), and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC using dichloromethane / methanol (30:1, v / v) elution to give 1H-indol-3-yl)-1H-bromoester C-12b (220 mg, 88% yield) as a yellow solid. LCMS: m / z (ESI), [M+H] + =724.23.

[0582] Step 2: At 70°C under a nitrogen atmosphere, a mixture of bromoester C-12b (120 mg, 0.17 mmol, 1 equivalent) and AgNO3 (56 mg, 0.34 mmol, 2 equivalents) in MeCN (15 mL) was stirred for 2 hours. The reaction is light-sensitive and should be avoided in the dark. The mixture was cooled to room temperature. The resulting mixture was filtered, and the filter cake was washed with CH2Cl2 (3 x 50 mL) and concentrated under reduced pressure to give nitrooxy ester C-12c (25 mg, 26% yield) as an orange solid. LCMS: m / z (ESI), [M+H]+ =707.2.

[0583] Step 3: At 0°C and under a nitrogen atmosphere, a mixture of nitrooxy ester C-12c (25 mg, 0.04 mmol, 1 equivalent) and TFA (0.6 mL, 8 mmol) in DCM (3 mL) was stirred for 30 minutes. The mixture was neutralized with ammonia to pH 7–8 and concentrated under reduced pressure. The residue was further purified by preparative HPLC under the following conditions (mobile phase consisting of water and acetonitrile containing 0.1% formic acid; constant gradient, mobile phase: 83% aqueous / 17% organic to 70% aqueous / 30% organic, 7 min; constant flow rate 70 mL / min) to give enantiomer 1 of Example 12 (2 mg, 9% yield) as a white solid. LCMS: m / z (ESI), [M+H] + =507.35. 1 H NMR: (DMSO-d6, 400MHz) δ1.92-1.96(2H,m),2.47-2.50(2H,m),3.03-3.07(2H,m),4.07-4.11(1H,m),4.54(2H,t ),5.09(2H,s),6.97-7.01(1H,m),7.37(3H,m),7.46(2H,d),7.59(2H,d),7.95(2H,s),8.28(1H,s),10.08(1H,br s),10.83(1H,s).

[0584] Example 14

[0585] Preparation of (R)-N-(3-(1H-pyrazol-4-yl)-1H-indol-7-yl)-3-amino-2-phenylpropionamide

[0586]

[0587] Step 1: Under nitrogen atmosphere and at 90 °C, dioxane (60.00 mL) containing amine I-7b (3.560 g, 8.63 mmol), tert-butyl 4-bromopyrazole-1-carboxylate (3.20 g, 12.95 mmol), potassium carbonate (3.58 g, 25.90 mmol), and PdCl₂dppf (705.11 mg, 863.43 μmol) and water (12.00 mL) was stirred for 16 hours. The solvent was removed under reduced pressure. The residue was purified by column chromatography using silica gel and eluting with a 10-50% gradient of ethyl acetate / petroleum ether. The purified fraction was evaporated to dryness to give amine I-14b (0.471 g, 748.56 μmol, 9% yield, 72% purity). LCMS:t R=1.77 min, 3 min chromatography (3 min - 5 - 95% MeCN / water (0.02% TFA), Waters Acquity UPLC BEH C18 1.7 μm, 2.1*50 mm, 40 °C), MS (ESI) m / z = 453.6 [M+H] + ; and amine I-14a as a brown solid (3.19 g, 6.88 mmol, 80% yield, 76% purity). LCMS: tR = 1.43 min, 3 min chromatogram (3 min - 5 - 95% MeCN / water (0.02% TFA), Waters AcquityUPLC BEH C18 1.7 μm, 2.1*50 mm, 40 °C), MS (ESI) m / z = 353.6 [M+H] + .

[0588] Step 2: At 0°C, a stirred mixture of amine I-14b (85 mg, 135.09 μmol), (R)-3-((tert-butoxycarbonyl)amino)-2-phenylpropionic acid (39.42 mg, 148.60 μmol), and DIPEA (26.19 mg, 202.64 μmol, 35.29 μL) in 2.00 mL of NMP was cooled and stirred. Then, at this temperature, HATU (56.80 mg, 149.39 μmol) was added in a single dose. The resulting mixture was stirred at room temperature for 17 hours. The reaction was then monitored by LCMS, and the reaction was found to be incomplete. More (R)-3-((tert-butoxycarbonyl)amino)-2-phenylpropionic acid (107.52 mg, 405.27 μmol) was added to the reactants. The reaction mixture was cooled and stirred at 0°C. At this temperature, a further addition of DIPEA (69.84 mg, 540.36 μmol, 94.12 μL) was made. The resulting mixture was stirred at room temperature for 31 hours. The solution was purified by passing it through a C18 fast column and eluting with a 0-80% gradient of MeCN / water (6 mmol / L NH4HCO3). The purified fraction was lyophilized to give amide(R)-enantiomer C-14c (15 mg, 16% yield) as a white solid. LCMS:t R =2.18 min, 3 min chromatography (3 min - 5 - 95% MeCN / water (0.02% TFA), Waters Acquity UPLC BEH C18 1.7um, 2.1*50mm, 40℃), MS (ESI) m / z = 700.5 [M+H] + .

[0589] Step 3: At 0 °C, a solution of (R)-enantiomer C-14c (15 mg, 21.43 μmol) in DCM (1.00 mL) was cooled and stirred in an ice / water bath. Then, 4 M HCl (72.76 mg, 2.00 mmol, 0.50 mL) was added dropwise to the reaction mixture. The resulting mixture was stirred at 0 °C for 1 hour. The reaction mixture was concentrated under reduced pressure. The residue was purified by C18 rapid chromatography with a 0% to 50% gradient elution of MeCN / water (6 mmol / L NH4HCO3). The purified fraction was lyophilized to give (R)-enantiomer C-14d (10 mg, 93% yield) as a white solid. LCMS:t R =1.59 min, 3 min chromatography (3 min - 5 - 95% MeCN / water (0.02% TFA), Waters Acquity UPLC BEH C18 1.7 μm, 2.1*50 mm, 40 °C), MS (ESI) m / z = 500.2 [M+H] + .

[0590] Step 4: A solution of (R)-enantiomer C-14d (10 mg, 20.02 μmol) was dissolved in THF containing 1 M TBAF (104.67 mg, 400.34 μmol, 0.40 mL). The reaction mixture was sealed and heated at 80 °C for 16 hours. The reaction solution was evaporated under reduced pressure. The residue was purified by passing it through a C18 rapid column and eluting with a CH3CN / water (0.02% TFA) gradient from 0 to 30%. The purified fraction was evaporated to dryness to give (R)-enantiomer Example 14 (4 mg, 8.73 μmol, 44% yield, TFA salt) as a brown solid. LCMS:t R =1.23 min, 3 min chromatography (3 min - 5 - 95% MeCN / water (0.02% TFA), Waters Acquity UPLC BEH C18 1.7 μm, 2.1*50 mm, 40 °C), MS (ESI) m / z = 346.3 [M+H] + . 1 HNMR: (400MHz, DMSO-d6) δ10.75(s,1H),10.20(s,1H),7.97(d,J=12.6Hz,5H),7.66–7.56(m,2H),7.48(d,J=7.4Hz,2H),7.42(t ,J=7.4Hz,2H),7.35(d,J=7.5Hz,2H),6.99(t,J=7.9Hz,1H),4.21(dd,J=9.4,4.6Hz,1H),3.58–3.55(m,1H),3.16–3.12(m,1H).

[0591] Example 15

[0592] Preparation of (S)-N-(3-(1H-pyrazol-4-yl)-1H-indol-7-yl)-3-amino-2-phenylpropionamide

[0593]

[0594] Step 1: At 90°C and under nitrogen atmosphere, nitro-indole I-15a (219 mg, 641.93 μmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxoboron-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole (249.81 mg, 770.32 μmol), potassium carbonate (266.16 mg, 1.93 mmol, 116.23 μL), and PdCl2dppf (52.42 mg, 64.19 μmol) were stirred in dioxane (5.00 mL) and water (1 mL) for 16 hours. The solvent was removed under reduced pressure. The residue was purified by silica gel column chromatography with a methanol / DCM gradient elution from 0 to 10%. The pure fraction was evaporated to dryness to give pyrazole I-15b (200 mg, 87% yield) as a brown solid. LCMS:t R =2.01 min, 3 min chromatography (3 min - 5 - 95% MeCN / water (0.02% TFA), Waters Acquity UPLC BEH C18 1.7um, 2.1*50mm, 40℃), MS (ESI) m / z = 359.3 [M+H] + .

[0595] Step 2: Iron powder (155.79 mg, 2.79 mmol) was added to a mixture of pyrazole I-15b (200 mg, 557.93 μmol) and ammonium hydrochloride (298.45 mg, 5.58 mmol) in ethanol (10 mL) and water (10 mL). The reaction mixture was then stirred and heated at 80 °C for 3 hours. The reaction mixture was filtered through diatomaceous earth and washed with ethanol (10 mL x 3), and the filtrate was concentrated. The residue was dissolved in 50 mL of ethyl acetate, filtered, and the filtrate was concentrated under reduced pressure to give amine I-15c (160 mg, 87% yield) as a brown solid. LCMS:t R =1.56 min, 3 min chromatography (3 min - 5 - 95% MeCN / water (0.02% TFA), Waters Acquity UPLC BEH C18 1.7 μm, 2.1*50 mm, 40 °C), MS (ESI) m / z = 329.3 [M+H] + .

[0596] Step 3: At 0 °C, a stirred mixture of (S)-3-((tert-butoxycarbonyl)amino)-2-phenylpropionic acid (40.38 mg, 152.21 μmol), amine I-15c (50 mg, 152.21 μmol), and DIPEA (59.02 mg, 456.64 μmol, 79.54 μL) in NMP (2 mL) was cooled and stirred. Then, at this temperature, HATU (69.82 mg, 183.63 μmol) was added in a single batch. The resulting mixture was stirred at 0 °C for 2 hours. The solution was purified by passing it through a C18 rapid column and eluting with a MeCN / water gradient from 0 to 60%. The purified fraction was lyophilized to give amide (S)-enantiomer C-15 (38 mg, 43% yield) as a white solid. LCMS:t R =2.08 min, 3 min chromatography (3 min - 5 - 95% MeCN / water (0.02% TFA), WatersAcquity UPLC BEH C18 1.7um, 2.1*50mm, 40℃), MS (ESI) m / z = 576.6 [M+H] + .

[0597] Step 4: At 0 °C, a solution of amide (S)-enantiomer C-15 (38 mg, 66.00 μmol) in DCM (2.00 mL) was cooled and stirred in an ice / water bath. Then, 4 M HCl (145.83 mg, 4.00 mmol, 1.00 mL) was added dropwise to the reaction mixture. The resulting mixture was stirred at 0 °C for 2 hours. The reaction mixture was concentrated under reduced pressure. The residue was purified by C18 rapid chromatography with a 0% to 50% gradient elution of MeCN / water (6 mmol / L NH4HCO3). The purified fraction was lyophilized to give (S)-enantiomer Example 15 (3.2 mg, 14% yield) as a white solid. LCMS:t R =1.17 min, 3 min chromatography (3 min - 5 - 95% MeCN / water (0.02% TFA), Waters Acquity UPLC BEH C18 1.7 μm, 2.1*50 mm, 40 °C), MS (ESI) m / z = 346.2 [M+H] + . 1H NMR: (400MHz, DMSO-d6) δ10.87(s,1H),7.95(s,2H),7.58(d,J=8.1Hz,2H),7.43(t,J=8.8Hz,3H),7.40–7.32( m,3H),7.29(t,J=7.1Hz,1H),6.98(t,J=7.7Hz,1H),4.07–3.97(m,1H),3.61–3.56(m,1H),3.00–2.93(m,1H).

[0598] Examples 16 (racemate), 17 (enantiomer 1), and 18 (enantiomer 2)

[0599] Preparation of N-(3-(1H-pyrazol-4-yl)-1H-indol-7-yl)-3-amino-2-(3-methoxyphenyl)propionamide

[0600]

[0601] Step 1: At room temperature, add 1.5 mL of CH3CN containing acid I-2 (237.58 mg, 0.80 mmol, 1.2 equivalents), TCFH (282.14 mg, 1.01 mmol, 1.5 equivalents), NMI (220.16 mg, 2.68 mmol, 4 equivalents), and amine I-49 (200 mg, 0.67 mmol, 1.00 equivalents) to a 20 mL vial. Stir the resulting mixture for 2 hours at room temperature under a nitrogen atmosphere. Concentrate the resulting mixture under reduced pressure. Purify the residue by preparative TLC with DCM / methanol (30:1, v / v) to give amide C-16a (150 mg, 38.87%) as a yellow solid. LCMS: m / z (ESI), [M+H] + =576.30.

[0602] Step 2: Purification of racemic amide C-16a was performed by preparative chiral HPLC under the following conditions: (Column: CHIRALPAK IG, 2*25cm, 5μm; Mobile phase A: Hex (0.2% DEA), Mobile phase B: EtOH:DCM = 1:1, v / v; Flow rate: 20 mL / min; Gradient: 25% B, 23 min; Wavelength: 220 / 254 nm; t R -1 = 14.66 minutes; t R -2=19.11 minutes; Sample solvent: EtOH:DCM=1:1, v / v; Injection volume: 0.3mL), to obtain enantiomers C-16b and C-16c as white solids.

[0603] Step 3: At room temperature, add 1 mL of TFA containing enantiomer C-16c (50 mg, 0.087 mmol, 1 equivalent) and DCM (2 mL) to a 50 mL round-bottom flask. Stir the resulting mixture at room temperature for 1.5 hours. Alkalinize the mixture to pH 9 with saturated NaHCO3 (aqueous solution). Extract the mixture with CH2Cl2 (3 x 10 mL). Concentrate the combined organic layers under reduced pressure. Purify the crude product by preparative HPLC under the following conditions: column: XBridge Prep OBD 30*150 mm, 5 μm; mobile phase A: water (10 mmol / L NH4HCO3), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 10% B to 40% B, 7 min; wavelength: 220 nm; t R =6.25 minutes), to obtain enantiomer 2 of Example 18 (14.0 mg, 42.93%) as a white solid. LCMS: m / z (ESI), [M+H) + =376.15. 1 H NMR(CD3OD,400MHz)δ3.05(1H,dd),3.43(1H,dd),3.83(3H,s),3.93(1H,dd),6.90(1H ,ddd),7.00–7.13(4H,m),7.26–7.35(1H,m),7.47(1H,s),7.64(1H,dd),7.91(2H,s). SFC:t R =3.248, ee = 97.14%.

[0604] At room temperature, 1 mL of TFA containing enantiomer C-16b (50 mg, 0.087 mmol, 1 equivalent) and DCM (2 mL) was added to a 50 mL round-bottom flask. The resulting mixture was stirred for 1.5 hours at room temperature. The mixture was alkalized to pH 7 with saturated NaHCO3 (aqueous solution). The mixture was extracted with CH2Cl2 (3 x 10 mL). The combined organic layers were concentrated under reduced pressure. The crude product was purified by preparative HPLC under the following conditions: (Column: XBridge Shield RP18 OBD column, 30*150 mm, 5 μm; Mobile phase A: water (10 mmol / L NH4HCO3 + 0.1% NH3·H2O), Mobile phase B: ACN; Flow rate: 60 mL / min; Gradient: 18% B to 40% B, 8 min; Wavelength: 220 nm; t R =7.52 minutes), to obtain enantiomer 1 of Example 17 (4.3 mg, 13.19%) as a white solid. LCMS: m / z (ESI), [M+H) + =376.15.1 H NMR(CD3OD,400MHz)δ3.05(1H,dd),3.43(1H,dd),3.83(3H,s),3.93(1H,dd),6.90(1H ,ddd),7.00–7.13(4H,m),7.26–7.35(1H,m),7.47(1H,s),7.64(1H,dd),7.91(2H,s). SFC:t R = 3.744 minutes, ee = 95.56%.

[0605] Step 4: At room temperature, add 2 mL of DCM containing racemic amide C-16a (20 mg, 0.035 mmol, 1 equivalent) and TFA (1 mL) to a 25 mL round-bottom flask. Stir the resulting mixture for 2 hours at room temperature. Alkalize the mixture to pH 9 with saturated NaHCO3. Extract the resulting mixture with CH2Cl2 (3 x 50 mL). Purify the crude product by preparative HPLC under the following conditions: column: XBridge Prep OBD C18 30*150 mm, 5 μm; mobile phase A: water (10 mmol / L NH4HCO3), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 14% B to 39% B, 8 min; wavelength: 220 nm; t R =6.97 minutes), to obtain the racemic amide of Example 16 (1.6 mg, 12.27%) as a white solid. LCMS: m / z (ESI), [M+H] + =376.20. 1 H NMR(CD3OD,400MHz)δ3.04–3.08(1H,m),3.32-3.50(1H,m),3.83(3H,s),3.93(2H,dd), 6.90(1H,ddd),6.99–7.18(4H,m),7.32(1H,t),7.47(1H,s),7.64(1H,dd),7.92(2H,s).

[0606] Examples 19 (racemate), 20 (enantiomer 1), and 21 (enantiomer 2)

[0607] Preparation of N-(3-(1H-pyrazol-4-yl)-1H-indol-7-yl)-3-amino-2-(3-cyanophenyl)propionamide

[0608]

[0609] Step 1: At 25°C under a N2 atmosphere, TCFH (366.78 mg, 1.31 mmol, 1.2 equivalents) and NMI (357.77 mg, 4.36 mmol, 4 equivalents) were added to a solution of acid I-3 (379.51 mg, 1.31 mmol, 1 equivalent) in CH3CN (20 mL). The mixture was stirred at 25°C for 2 hours. The resulting mixture was filtered, and the filter cake was washed with methanol (3 x 100 mL). The filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC with PE / EA (1:1, v / v) to give a racemic amide C-20a (350 mg, 54.46%) as a brown solid. LCMS: m / z (ESI), [M+H] + =571.30.

[0610] Step 2: Purification of racemic amide C-20a (350 mg, 0.61 mmol, 1 equivalent) by chiral HPLC under the following conditions: (Column: CHIRALPAK IG, 2*25 cm, 5 μm; Mobile phase A: Hex (0.5% 2M NH3-methanol), Mobile phase B: EtOH:DCM (1:1, v / v); Flow rate: 20 mL / min; Gradient: 40% B to 40% B, 8.5 min; Wavelength: 220 / 254 nm; t R -1 = 5.60 minutes; t R -2 = 7.14 minutes. Sample solvent: EtOH:DCM (1:1, v / v); injection volume: 0.6 mL, to obtain enantiomer amide C-20c (112 mg, 31.68%) as a white solid. LCMS m / z (ESI), [M+H) + =571.30. Chiral HPLC:t R =1.336 minutes, ee=100%, and the enantiomer amide C-20b (119 mg, 33.66%) was a white solid. LCMS: m / z (ESI), [M+H] + =571.30. Chiral HPLC:t R = 1.735 minutes, ee = 100%.

[0611] Step 3: At 25°C under a N2 atmosphere, TFA (5 mL) was added to a solution of enantiomer amide C-20c (102 mg, 0.18 mmol, 1 equivalent) in DCM (15 mL). The mixture was stirred at 25°C for 2 hours. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in methanol (50 mL). The mixture was alkalized to pH 8 with NH3 (aqueous solution). The resulting mixture was concentrated under vacuum. The crude product was purified by preparative HPLC under the following conditions (column: XBridge Shield RP18 OBD column, 30*150 mm, 5 μm; mobile phase A: water 10 mmol / L NH4HCO3), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 13% B to 39% B, 8 min; wavelength: 220 nm; t R =7.58 minutes), to obtain enantiomer 2 of Example 21 as a white solid (51.4 mg, 76.95%). LCMS m / z (ESI), [M+H) + =371.10. 1 H NMR(CD3OD,400MHz)δ3.03(1H,m),3.42(1H,m),3.98(1H,dt),7.06(2H,m),7 .47(1H,s),7.58(1H,m),7.67(2H,m),7.80(1H,m),7.86(1H,t),7.91(2H,s). SFC:t R =2.432 minutes, ee = 98.42%.

[0612] At 25°C under a nitrogen atmosphere, TFA (5 mL) was added to a solution of enantiomer amide C-20b (109 mg, 0.19 mmol, 1 equivalent) in DCM (15 mL). The mixture was stirred at 25°C for 2 hours. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in methanol (50 mL). The mixture was alkalized to pH 8 with NH3 (aqueous solution). The resulting mixture was concentrated under vacuum. The crude product was purified by preparative HPLC under the following conditions (column: XBridge Shield RP18 OBD column, 30*150 mm, 5 μm; mobile phase A: water (10 mmol / L NH4HCO3), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 13% B to 39% B, 8 min; wavelength: 220 nm; t R =7.57 minutes), to obtain enantiomer 1 of Example 20 (37.4 mg, 52.10%) as a white solid. LCMS m / z (ESI), [M+H] + =371.10. 1H NMR(CD3OD,400MHz)δ3.04(1H,dd),3.43(1H,dd),3.98(1H,dd),7.06(2H,m),7 .47(1H,s),7.58(1H,t),7.68(2H,m),7.81(1H,dt),7.87(1H,d),7.92(2H,s). SFC:t R =1.999 minutes, ee=100%.

[0613] Step 4: At room temperature and under air atmosphere, the mixture of TFA (2 mL) and racemic amide C-20a (170 mg, 0.30 mmol, 1 equivalent) in DCM (6 mL) was stirred for 1 hour. The resulting mixture was concentrated under reduced pressure. The resulting mixture was diluted with water (20 mL). The mixture was alkalized to pH 8 with saturated NaHCO3 (aqueous solution). The resulting mixture was extracted with ethyl acetate (3 x 20 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The crude product (100 mg) was purified by preparative HPLC under the following conditions to give the racemic solid Example 19 (30 mg, 26.68%) as a white solid. LCMS: m / z (ESI), [M+H] + =371.10. 1 H NMR(400MHz,DMSO-d6)δ3.06(1H,d),3.45(1H,d),4.01(1H,d),7.02–7.10(2H,m),7.4 7(1H,s),7.59(1H,t),7.62–7.68(1H,m),7.71(1H,t),7.82(1H,t),7.86–7.94(3H,m).

[0614] Example 22 (racemic mixture)

[0615] Preparation of 3-amino-2-cyclohexyl-N-[3-(1H-pyrazol-4-yl)-1H-indol-7-yl]propionamide (racemate)

[0616]

[0617] Step 1: At room temperature and under a nitrogen atmosphere, HATU (190.99 mg, 0.50 mmol, 1.5 equivalent) and DIEA (129.84 mg, 1.01 mmol, 3 equivalent) were added fractionally to a stirred solution of amine I-50 (110 mg, 0.34 mmol, 1 equivalent) and 3-[(tert-butoxycarbonyl)amino]-2-cyclohexylpropionic acid (99.95 mg, 0.37 mmol, 1.1 equivalent) in DMF (5 mL). The resulting mixture was stirred for 2 hours at room temperature and under a nitrogen atmosphere. The crude product was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water, 0% to 100% gradient, 20 min; detector, UV 254 nm. The resulting mixture was concentrated under vacuum. This gave the racemic amide C-22a (70 mg, 35.93%) as a white solid. LCMS:m / z(ESI),[M+H] + =582.50.

[0618] Step 2: At room temperature and under a nitrogen atmosphere, a solution of TBAF (7M) containing racemic amide C-22a (40 mg, 0.069 mmol, 1 equivalent) in THF (2 mL) was stirred overnight. The resulting mixture was diluted with water (10 mL). The mixture was extracted with DCM (3 x 20 mL). The combined organic layers were dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC with DCM / methanol (15:1, v / v) to give pyrazole C-22b (15 mg, 48.32%) as a white solid. LCMS: m / z (ESI), [M+H] + =452.30.

[0619] Step 3: At room temperature and under a nitrogen atmosphere, a solution of pyrazole C-22b (15 mg, 0.033 mmol, 1 equivalent) and TFA (0.5 mL) in DCM (1 mL) was stirred for 2 hours. The crude product was purified by preparative HPLC under the following conditions: column: XBridge Prep OBD 30*150 mm, 5 μm; mobile phase A: water (10 mmol / L NH4HCO3), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 17% B to 42% B, 8 min; wavelength: 254 nm; t R =7.47 minutes), to obtain example 22 (2 mg, 16.57%), a racemic mixture appearing as a white solid. LCMS: m / z (ESI), [M+H) + =352.20. 1H NMR(400MHz,DMSO-d6)δ1.06–0.95(1H,m),1.31–1.10(4H,m),12.81(1H,s),1.91–1.50(6H,m),2.38(1H,td ),2.96–2.81(2H,m),7.00(1H,t),7.43(1H,d),7.62–7.53(2H,m),7.95(2H,s),9.76(1H,s),10.92(1H,s).

[0620] Examples 23 (enantiomer 1) and 24 (enantiomer 2)

[0621] Preparation of N-(3-(1H-pyrazol-4-yl)-1H-indol-7-yl)-3-amino-2-cyclohexylpropionamide

[0622]

[0623] Step 1: At room temperature, amine I-49 (300 mg, 1.01 mmol, 1 equivalent) and 3-[(tert-butoxycarbonyl)amino]-2-cyclohexylpropionic acid (300.15 mg, 1.11 mmol, 1.1 equivalent), TCFH (338.56 mg, 1.21 mmol, 1.2 equivalent), NMI (330.24 mg, 4.02 mmol, 4 equivalent), and MeCN (5 mL) were added to a 40 mL vial. The mixture was stirred for 1.5 h at room temperature under a nitrogen atmosphere. The reaction was monitored by LCMS. The desired product was detected by LCMS. The mixture was concentrated under reduced pressure. The residue was purified by preparative TLC (CH2Cl2 / methanol 45:1) to give amide C-23a (420 mg, 75.71%) as a yellow solid. LCMS: m / z (ESI), [M+H] + =552.40.

[0624] Step 2: Amide C-23a (420 mg) was purified by preparative chiral HPLC under the following conditions: (Column: CHIRALPAK IE, 2*25 cm, 5 μm; Mobile phase A: MTBE (0.1% DEA), Mobile phase B: ACN (0.1% IPA); Flow rate: 20 mL / min; Gradient: 10% B to 10% B, 16 min; Wavelength: 220 / 254 nm; t) R -1 = 8.86 minutes; t R-2 = 12.61 minutes; Sample solvent: IPA; Injection volume: 0.6 mL), to obtain enantiomer C-23c and enantiomer C-23b (170 mg, 40.5%) as a yellow solid. LCMS: m / z (ESI), [M+H) + =552.40.

[0625] Step 3: At room temperature, add amide C-23c (200 mg, 0.36 mmol, 1 equivalent), TFA (1 mL), and DCM (3 mL) to a 100 mL round-bottom flask. Stir the resulting mixture for 1 hour at room temperature under air. Concentrate the resulting mixture under reduced pressure. Dilute the resulting mixture with DMF (2 mL). Alkalize the mixture to pH 9 with NH3 (aqueous solution). Stir the resulting mixture for 30 minutes at 0 °C under air. Purify the crude product (180 mg) by preparative HPLC under the following conditions: (Column: XBridge Shield RP18 OBD 30*150 mm, 5 μm; Mobile phase A: Water (10 mmol / L NH4HCO3 + 0.1% NH3 aqueous solution), Mobile phase B: ACN; Flow rate: 60 mL / min; Gradient: 20% B to 45% B, 8 min; Wavelength: 220 nm; t R =7.67 minutes), to obtain enantiomer 2 of Example 24 (69.4 mg, 53.98%) as a white solid. LCMS: m / z (ESI), [M+H) + =352.15. 1 H NMR(DMSO-d6,400MHz)δ0.98(1H,q),1.18(4H,p),1.64(5H,d),1.83(1H,d),2.37(1 H,s),2.87(1H,m),3.22(1H,s),7.00(1H,m),7.44(1H,d),7.57(2H,m),7.95(2H,s). SFC:t R = 1.971 minutes, ee = 97.42%.

[0626] At room temperature, amide C-23b (170 mg, 0.31 mmol, 1 equivalent), TFA (1 mL), and DCM (3 mL) were added to a 100 mL round-bottom flask. The mixture was stirred for 1 hour at room temperature under air. The mixture was then concentrated under reduced pressure. The mixture was diluted with DMF (2 mL). The mixture was alkalized to pH 9 with NH3 (aqueous solution). The mixture was stirred for 30 minutes at 0 °C under air. The crude product (150 mg) was purified by preparative HPLC under the following conditions: column: YMC-Actus Triart C18 ExRS, 30*150 mm, 5 μm; mobile phase A: water (10 mmol / L NH4HCO3), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 18% B to 43% B, 7 min; wavelength: 220 nm; t R =6.55 minutes), to obtain enantiomer 1 of Example 23 (57 mg, 52.26%) as a white solid. LCMS: m / z (ESI), [M+H) + =352.20. 1 H NMR (DMSO-d6, 400MHz) δ1.21(5H,m),1.75(6H,dd),2.35(1H,d),2.86(1H,m),3.21(1H,s),7.00(1H,m),7.44(1H,m),7.57(2H,m),7.95(2H,s). SFC:t R = 1.860 minutes, ee = 100%.

[0627] Examples 26 (enantiomer 1) and 27 (enantiomer 2)

[0628] Preparation of N-(3-(1H-pyrazol-4-yl)-1H-indol-7-yl)-3-amino-2-(tetrahydro-2H-pyran-4-yl)propionamide

[0629]

[0630] Step 1: At room temperature, 3-[(tert-butoxycarbonyl)amino]-2-(oxacyclohexane-4-yl)propionic acid (250 mg, 0.92 mmol, 1 equivalent), amine I-49 (272.88 mg, 0.92 mmol, 1 equivalent), TCFH (513.26 mg, 1.83 mmol, 2 equivalents), NMI (375.49 mg, 4.58 mmol, 5 equivalents), and CH3CN (10 mL) were added to a 40 mL vial. The resulting mixture was stirred at room temperature for 2 hours. The residue was purified by preparative TLC (CH2Cl2 / methanol (25:1) to give a brown solid racemic amide C-26a (312 mg, 59.76%). LCMS: m / z (ESI), [M- t Bu+H] + =498.25.

[0631] Step 2: Purification of racemic amide C-26a (312 mg, 59.76%) was performed by preparative chiral HPLC under the following conditions: (Column: CHIRALPAK IF, 2*25 cm, 5 μm; Mobile phase A: Hex (0.2% DEA), Mobile phase B: EtOH:DCM (1:1, v / v); Flow rate: 20 mL / min; Gradient: 30% B to 30% B, 12 min; Wavelength: 220 / 254 nm; t R -1 = 7.46 minutes; t R -2 = 9.02 minutes; Sample solvent: EtOH:DCM (1:1, v / v); Injection volume: 0.2 mL), to obtain enantiomer C-26c (130 mg, 41.6%), SFC:t R =1.932 minutes, ee=100%, and the enantiomer C-26b (120 mg, 38.4%) as a white solid, SFC:t R =2.524 minutes, ee = 99.634%.

[0632] Step 3: At room temperature, add enantiomer C-26c (120 mg, 0.22 mmol, 1 equivalent) and DCM (3 mL) to a 5...

Claims

1. A compound of formula I or a pharmaceutically acceptable salt thereof: in: R 1 and R 2 each independently selected from the group consisting of hydrogen, hydroxy, halogen, cyano, amino, alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, and heteroalkynyl, wherein said alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, and heteroalkynyl are each optionally substituted with one or more groups independently selected from the group consisting of hydroxy, halogen, cyano, or amino; X, W, Z and U are each independently N or C(R 3 ); R 3 selected from the group consisting of hydrogen, hydroxy, halogen, cyano, amino, alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, and heteroalkynyl, wherein said alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, and heteroalkynyl are each optionally substituted with one or more groups independently selected from the group consisting of hydroxy, halogen, cyano, or amino; E is-N(R 4 )C(=O)-, -C(=O)N(R 4 )-, -N(R 4 )SO2-or-SO2N(R 4 )-; R 4 is selected from the group consisting of: hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, and heteroalkynyl; Y is -Y 1 -Y 2 -(Y 3 ) n , where Y 1 Does not exist or is -C(R 5 )2-, Each R 5 are independently selected from the group consisting of: hydrogen, -N(R a )2, alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl and -alkyl-heterocyclyl, wherein the alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl and -alkyl-heterocyclyl are optionally substituted with one or more R 6 replace, Each R 6 are independently selected from the group consisting of hydroxy, halogen, cyano, amino, alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, and -alkyl-N(R a )2; Each R a are independently selected from hydrogen or alkyl; Y 2 Selected from –(CH2) p -cycloalkyl-*, –(CH2) p -Heterocyclyl-*, –(CH2) p -Aryl-* or –(CH2) p -heteroaryl-*, each of which is optionally substituted with one or more groups independently selected from the group consisting of hydroxy, halogen, cyano, amino, alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl or heteroalkynyl, and wherein p is 0, 1 or 2, and Y 2 The * end and Y 3 connected; Y 3 Does not exist or -Y 31 -Y 32 -Y 33 ;in Y 31 is selected from absent, alkyl, alkenyl or alkynyl; Y 32 is selected from the group consisting of absent, -O-#, -OC(=O)-#, -C(=O)O-#, -P(=O)(R b )-#、-OC(=O)N(R b )-#、-N(R b )C(=O)-# or -C(=O)N(R b )-#, where each R b are independently selected from hydrogen or alkyl, and Y 32 The # end and Y 33 connect; Y 33 Selected from the group consisting of: hydrogen, hydroxyl, cyano, halogen, -N(R c )2, alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, wherein the alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are optionally substituted with one or more R 7 Substitute, where each R 7 is independently selected from halogen, hydroxy, amino, cyano, nitrooxy, alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl or haloalkyl, and each R c are independently selected from hydrogen or alkyl; and n is an integer of 1-5.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R 1 and R 2 Each is independently hydrogen, halogen, hydrogen, cyano, amino or alkyl optionally substituted with one or more halogens.

3. The compound according to claim 2 or a pharmaceutically acceptable salt thereof, wherein R 1 is hydrogen or alkyl, and R 2 is hydrogen, halogen or alkyl.

4. The compound according to claim 3 or a pharmaceutically acceptable salt thereof, wherein R 1 and R 2 It's all hydrogen.

5. The compound according to claim 3 or a pharmaceutically acceptable salt thereof, wherein R 1 and R 2 All are alkyl.

6. The compound according to claim 5 or a pharmaceutically acceptable salt thereof, wherein R 1 is a methyl group, and R 2 It's methyl.

7. The compound according to claim 3 or a pharmaceutically acceptable salt thereof, wherein R 1 is hydrogen, and R 2 It's a halogen.

8. The compound according to claim 7 or a pharmaceutically acceptable salt thereof, wherein R 1 is hydrogen, and R 2 It's fluorine.

9. The compound according to claim 3 or a pharmaceutically acceptable salt thereof, wherein R 1 is hydrogen, and R 2 It is an alkyl group.

10. The compound according to claim 9 or a pharmaceutically acceptable salt thereof, wherein R 1 is hydrogen, and R 2 It's methyl.

11. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein X, W, U and Z are C(R 3 ).

12. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein X and U are C(R 3 ), W and Z are N.

13. A compound according to any one of claims 10 to 12 or a pharmaceutically acceptable salt thereof, wherein R 3 It's hydrogen.

14. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein E is -N(R 4 )C(=O)-or-C(=O)N(R 4 )-.

15. The compound according to claim 14 or a pharmaceutically acceptable salt thereof, wherein R 4 It's hydrogen.

16. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein Y 1 Does not exist.

17. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein Y 1 Yes-C(R 5 )2-.

18. The compound according to claim 17 or a pharmaceutically acceptable salt thereof, wherein one R 5 is hydrogen, and the other R 5 Selected from -N(R a )2, alkyl, cycloalkyl, heterocyclyl or -alkyl-heterocyclyl, wherein the alkyl, cycloalkyl, heterocyclyl and -alkyl-heterocyclyl are optionally substituted by one or more R 6 replace.

19. The compound according to claim 17 or a pharmaceutically acceptable salt thereof, wherein one R 5 is an alkyl group, and the other R 5 Selected from -N(R a )2, alkyl, cycloalkyl or heterocyclic, wherein the alkyl, cycloalkyl and heterocyclic are optionally substituted by one or more R 6 replace.

20. The compound according to claim 18 or 19, or a pharmaceutically acceptable salt thereof, wherein R 6 Selected from amino, alkyl or -alkyl-N(R a )2.

21. A compound according to any one of claims 17 to 20 or a pharmaceutically acceptable salt thereof, wherein Y 1 Select from the group consisting of:

22. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein Y 2 Yes – (CH2) p -cycloalkyl-*.

23. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein Y 2 Yes – (CH2) p -(C 3-10 cycloalkyl)-*, and p is 0 or 1.

24. The compound according to claim 23 or a pharmaceutically acceptable salt thereof, wherein Y 2 It is cyclopentyl or cyclohexyl.

25. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein Y 2 Yes – (CH2) p -heterocyclyl-*.

26. The compound according to claim 25 or a pharmaceutically acceptable salt thereof, wherein Y 2 Yes – (CH2) p -(5- to 12-membered heterocyclyl)-*, and p is 0 or 1.

27. The compound according to claim 26 or a pharmaceutically acceptable salt thereof, wherein Y 2 It is tetrahydrofuranyl, pyrrolidinyl or tetrahydropyranyl.

28. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein Y 2 Yes – (CH2) p -Aryl-*.

29. The compound according to claim 28 or a pharmaceutically acceptable salt thereof, wherein Y 2 Yes – (CH2) p -(C 5-12 aryl)-*, and p is 0 or 1.

30. The compound according to claim 29 or a pharmaceutically acceptable salt thereof, wherein Y 2 It is phenyl or -CH2-phenyl-*.

31. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein Y 2 Yes – (CH2) p -heteroaryl-*.

32. The compound according to claim 31 or a pharmaceutically acceptable salt thereof, wherein Y 2 Yes – (CH2) p -(5- to 12-membered heteroaryl)-*, and p is 0 or 1.

33. The compound according to claim 32 or a pharmaceutically acceptable salt thereof, wherein Y 2 It is pyridyl, tetrahydroisoquinolyl, -CH2-imidazolyl-* or -CH2-indolyl-*.

34. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein Y 31 Not present or alkyl.

35. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein Y 31 Does not exist.

36. The compound according to claim 35 or a pharmaceutically acceptable salt thereof, wherein Y 32 is selected from the group consisting of absent, -O-#, -C(=O)O-#, -P(=O)(R b )-#、-N(R b )C(=O)-# or -C(=O)N(R b )-#.

37. A compound according to claim 35 or 36, or a pharmaceutically acceptable salt thereof, wherein Y 33 is selected from hydrogen, hydroxy, cyano, halogen, -N(R c )2, alkyl or heteroalkyl, wherein the alkyl and heteroalkyl are optionally substituted by one or more R 7 replace.

38. The compound according to claim 37 or a pharmaceutically acceptable salt thereof, wherein Y 33 Selected from hydrogen, hydroxy, cyano, halogen, -NH2, methyl, -CH2CH2OCH3.

39. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein Y 31 It is an alkyl group.

40. The compound according to claim 38 or a pharmaceutically acceptable salt thereof, wherein Y 32 is selected from the group consisting of absent, -O-#, -OC(=O)-#, -OC(=O)N(R b )-# or -N(R b )C(=O)-#.

41. The compound according to claim 38 or 39, or a pharmaceutically acceptable salt thereof, wherein Y 33 Selected from -N(R c )2, alkyl or aryl, wherein the alkyl and aryl groups are optionally substituted by one or more R 7 replace.

42. The compound according to claim 41 or a pharmaceutically acceptable salt thereof, wherein each R 7 Independently selected from halogen, hydroxy, amino, cyano, nitrooxy or alkyl.

43. The compound according to claim 42 or a pharmaceutically acceptable salt thereof, wherein Y 33 Selected from -NH2, -N(CH3)2, methyl, dimethylphenyl or nitrooxypentyl.

44. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein n is 1 or 2.

45. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of:

46. ​​A pharmaceutical composition comprising a compound according to any one of claims 1 to 45 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

47. The pharmaceutical composition according to claim 46, which is formulated as eye drops.

48. A method of inhibiting ROCK activity in a subject in need thereof, comprising administering to the subject an effective amount of a compound according to any one of claims 1 to 45 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 46 or 47.

49. A method of treating a ROCK-related disorder in a subject in need thereof, comprising administering to the subject an effective amount of a compound according to any one of claims 1 to 45 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 46 or 47.

50. The method of claim 49, wherein the ROCK-related disorder is an eye disease, including glaucoma and retinal diseases, such as wet AMD, dry AMD, and DME; a bone disorder, including osteoporosis; a vascular disease, including cerebral vasospasm, coronary vasospasm, hypertension, pulmonary hypertension, sudden death syndrome, angina pectoris, myocardial infarction, restenosis, stroke, hypertensive vascular disease, heart failure, cardiac graft vasculopathy, vein graft disease; a pulmonary disease, including chronic obstructive pulmonary disease (COPD) and asthma; a neurological disorder, including spinal cord injury, Alzheimer's disease, multiple sclerosis, depression, attention deficit hyperactivity disorder, and neuropathic pain; a neovascular disorder and cancer; obesity; and erectile dysfunction.

51. The method of claim 50, wherein the ROCK-related disorder is glaucoma.

52. The method of any one of claims 49 to 51, wherein the compound is administered simultaneously, separately or sequentially with one or more additional therapeutic agents.

53. The method of claim 52, wherein the one or more additional therapeutic agents are selected from beta blockers, alpha agonists, carbonic anhydrase inhibitors, prostaglandin-like compounds, miotic agents, or cholinergic or adrenergic compounds.

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