ROCK inhibitor and its use

Abstract

Compounds of formula (I) and pharmaceutically acceptable salts thereof, which are useful as ROCK inhibitors, are disclosed herein. Also disclosed are pharmaceutical compositions comprising a compound of formula (I), and methods of using such compounds or compositions for treating ROCK-related disorders (such as glaucoma).

Description

Technical Field

[0001]

[0001] The present disclosure generally relates to novel compounds that inhibit Rho - associated protein kinase ( "ROCK") and pharmaceutically acceptable salts thereof. The present disclosure also relates to pharmaceutical compositions containing the compounds as active ingredients and the use of the compounds in the treatment of ROCK - related disorders including glaucoma.

Background Art

[0002]

[0002] Rho - associated protein kinase (ROCK) is a member of the serine - threonine protein kinase family. ROCK exists in two isoforms, namely ROCK1 and ROCK2. Both isoforms are activated by the GTP - bound form of Rho GTPase. ROCK plays an important role in many cellular processes including smooth muscle cell contraction, cell proliferation, cell adhesion, and cell migration. Inhibiting ROCK activity has shown potential for therapeutic applications in a wide range of pathological conditions.

[0003]

[0003] Currently available ROCK inhibitors include Eril (for the treatment of cerebral vasospasm) from Asahi Kasei, Granatine (for the treatment of ocular hypertension and glaucoma) from Kowa, and Rhopressa (for reducing intraocular pressure (IOP) elevation in patients with open - angle glaucoma or ocular hypertension) from Aerie.

Summary of the Invention

Problems to be Solved by the Invention

[0004]

[0004] Therefore, novel compounds that inhibit ROCK are needed as pharmacological tools and have attracted considerable interest as drugs for treating ROCK - related disorders such as glaucoma.

Means for Solving the Problems

[0005] A novel compound capable of inhibiting ROCK is disclosed herein. As a result, the compounds of the present disclosure are useful for the treatment of ROCK-related diseases such as glaucoma.

[0006] In one aspect, the present disclosure provides a compound of formula (I):

[0006]

Chemical formula

[0007] [Wherein, Each of R 1 and R 2 is independently selected from the group consisting of hydrogen, hydroxy, halogen, cyano, amino, alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, and heteroalkynyl, and said alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, and heteroalkynyl are each optionally substituted with one or more groups independently selected from hydroxyl, halogen, cyano or amino, Each of X, W, Z, and U is independently N or C(R 3 ), R 3 is selected from the group consisting of hydrogen, hydroxy, halogen, cyano, amino, alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, and heteroalkynyl, and said alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, and heteroalkynyl are each optionally substituted with one or more groups independently selected from hydroxyl, halogen, cyano or amino, E is -N(R 4 )C(=O)-, -C(=O)N(R 4 )-, -N(R 4 )SO 2 -, or -SO 2 N(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 and, in the formula, Y 1 either does not exist or is -C(R 5 ) 2 -, each R 5 is 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, and said alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl and -alkyl-heterocyclyl are optionally substituted with one or more R 6 s, each R 6 is independently selected from the group consisting of hydroxy, halogen, cyano, amino, alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, and -alkyl-N(R a ) 2 s, each R a is independently selected from hydrogen or alkyl, Y 2 is selected from -(CH 2 ) p -cycloalkyl-*, -(CH 2 ) p -heterocyclyl-*, -(CH 2 ) p -aryl-*, or -(CH 2 ) p -heteroaryl-*, each of which may be optionally substituted with one or more groups independently selected from hydroxy, halogen, cyano, amino, alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, or heteroalkynyl, p is 0, 1 or 2, and the * end of Y 2 is Y3 is attached to Y 3 is absent or -Y 31 -Y 32 -Y 33 wherein, Y 31 is absent or is selected from alkyl, alkenyl or alkynyl, Y 32 is absent or is selected from -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 )-#, each R b is independently selected from hydrogen or alkyl, the # end of Y 32 is attached to Y 33 and Y 33 is hydrogen, hydroxyl, cyano, halogen, -N(R c ) 2 , alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl, and the alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl are optionally substituted with one or more R 7 , each R 7 is independently selected from halogen, hydroxy, amino, cyano, nitrooxy, alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, or haloalkyl, each R c is independently selected from hydrogen or alkyl, and n is an integer from 1 to 5] and a pharmaceutically acceptable salt thereof.

[0008]

[0007] In another aspect, the present disclosure provides a pharmaceutical composition comprising a compound of the present disclosure or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

[0008] In a further aspect, the present disclosure provides a method of inhibiting ROCK activity in a subject in need of inhibition of ROCK activity, the method comprising administering to the subject an effective amount of a compound of the present disclosure or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of the present disclosure.

[0009]

[0009] In a further aspect, the present disclosure provides a method of treating an ROCK-related disorder, the method comprising administering to a subject in need of treatment of the ROCK-related disorder an effective amount of a compound of the present disclosure or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of the present disclosure.

[0010]

[0010] In another aspect, the present disclosure provides the use of a compound of the present disclosure or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of the present disclosure in the manufacture of a medicament for treating an ROCK-related disorder.

[0011]

[0011] In another aspect, the present disclosure provides a compound of the present disclosure or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of the present disclosure for use in the treatment of an ROCK-related disorder.

BEST MODE FOR CARRYING OUT THE INVENTION

[0012]

[0012] Certain embodiments of the present disclosure are referred to in detail below, and examples of those embodiments are illustrated in the accompanying structures and formulas. The present disclosure is described in relation to the recited embodiments, but of course, those embodiments do not limit the present disclosure to those embodiments. Instead, the present disclosure is intended to embrace all alternatives, modifications, and equivalents that may be included within the scope of the present disclosure as defined by the claims. Those skilled in the art will recognize many methods and materials similar or equivalent to those described herein that may be used in the practice of the present disclosure. The present disclosure is in no way limited to the methods and materials described. If one or more of the incorporated references and similar materials, including defined terms, term usage, or described techniques, differ from or conflict with the present application, the present disclosure prevails. All references, patents, and patent applications cited in the present disclosure are hereby incorporated by reference in their entirety.

[0013]

[0013] Of course, certain features of the present disclosure are described in the context of separate embodiments for clarity, but may also be provided in combination within a single embodiment. Conversely, various features of the present disclosure are described in the context of a single embodiment for brevity, but may also be provided separately or in any suitable sub-combination. Also, as used in this specification and the appended claims, the singular forms "a," "an," and "the" include the plural unless the context clearly dictates otherwise. Thus, for example, the expression "a compound" includes a plurality of compounds. Definitions

[0014] The definitions of certain functional groups and chemical terms are described in more detail below. For the purposes of this disclosure, chemical elements are identified according to the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75th Edition, inside cover, and specific functional groups are generally defined as described therein. Further, general principles of organic chemistry, as well as specific functional moieties and reactivities, are described in Organic Chemistry, Thomas Sorrell, University Science Books, Sausalito, 1999; Smith and March, March’s Advanced Organic Chemistry, 5th Edition, John Wiley & Sons, Inc., New York, 2001; Larock, Comprehensive Organic Transformations, VCH Publishers, Inc., New York, 1989; Carruthers, Some Modern Methods of Organic Synthesis, 3rd Edition, Cambridge University Press, Cambridge, 1987, the entire contents of each of which are incorporated herein by reference.

[0014]

[0015] At various places in this disclosure, linking substituents are described. When the structure clearly requires a linking group, the Markush variables listed for that group are understood to be linking groups. For example, if the structure requires a linking group and the definition of the Markush group of that variable lists "alkyl", "alkyl" is understood to represent a linking alkylene group.

[0015]

[0016] If any variable (e.g., R i ) occurs two or more times in any component or formula for a compound, its definition at each occurrence is independent of its definition at all other occurrences. Thus, for example, if a group is shown to be substituted with from 0 to 2 R i moieties, the group is optionally substituted with up to 2 R imay be replaced in part, and R at each occurrence i is independently selected from the definition of R i . Combinations of substituents and / or variables are permitted, but only if such combinations result in stable compounds.

[0016]

[0017] As used herein, for convenience, a dash “-” is used before or after a chemical group to indicate the point of attachment of the substituent. For example, -OH is attached via an oxygen atom. That is, a chemical group can be represented with or without one or more dashes without losing its ordinary meaning. A wavy line drawn through a line in a structure indicates the point of attachment of a group. Unless chemically or structurally required, the order in which chemical groups are written or named does not indicate or imply directionality. As used herein, a solid line extending from the center of a ring indicates that the point of attachment of a substituent of that ring can be at any ring atom. When a substituent is listed without indicating the atom through which the substituent is attached to the remainder of a compound of a given formula, such substituent can be attached through any atom in such formula. Combinations of substituents and / or variables are permitted, but only if such combinations result in stable compounds.

[0017]

[0018] The recitation of a range of values herein is merely for convenience and serves to list separately each individual value that falls within the range, as if each individual value were separately recited herein. Each individual value is incorporated herein as if it were separately recited herein. The ranges used herein include the two limiting values of the range unless otherwise specified. For example, the expressions “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”.

[0018]

[0019] As used herein, the terms "compounds provided herein", "compounds disclosed herein", or "compounds of the present disclosure" refer to compounds of formula (I) and specific compounds disclosed herein.

[0019]

[0020] As used herein, the term "C i~j " indicates a range of carbon atom numbers, where i and j are integers, the range of carbon atom numbers includes both endpoints (i.e., i and j), and each integer point therebetween, and j is greater than i. By way of example, C 1~6 indicates a range of 1 to 6 carbon atoms and includes 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, and 6 carbon atoms. In some embodiments, the term "C 1~12 " indicates 1 to 12, particularly 1 to 10, particularly 1 to 8, particularly 1 to 6, particularly 1 to 5, particularly 1 to 4, particularly 1 to 3, or particularly 1 to 2 carbon atoms.

[0020]

[0021] As used herein, the term "alkyl", whether used as part of another term or independently, refers to a saturated straight-chain or branched-chain hydrocarbon radical and may be independently optionally substituted with one or more of the following substituents. The term "C i~j alkyl" refers to an alkyl 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. Examples of "C 1~10 alkyl" include, but are not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, and decyl. "C 1~6Examples of "alkyl" include, but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, i-butyl, s-butyl, t-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, and 3,3-dimethyl-2-butyl.

[0021]

[0022] As used herein, the term "alkenyl", whether used as part of another term or independently, refers to a straight or branched chain hydrocarbon radical having at least one carbon-carbon double bond, which may be independently optionally substituted with 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, ethenyl (or vinyl), propenyl (allyl), butenyl, pentenyl, 1-methyl-2-buten-1-yl, and 5-hexenyl.

[0022]

[0023] As used herein, the term "alkynyl", whether used as part of another term or independently, refers to a straight-chain or branched-chain hydrocarbon radical having at least one carbon-carbon triple bond and may be optionally independently substituted with one or more substituents described herein. In some embodiments, the alkynyl 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, and 2-propynyl.

[0023]

[0024] As used herein, the term "amino" refers to an -NH 2 group. The amino group may also be substituted with one or more groups such as an alkyl group, an aryl group, a carbonyl group or another amino group.

[0024]

[0025] As used herein, the term "aryl", whether used as part of another term or independently, refers to monocyclic and polycyclic ring systems having a total of 5 to 20 ring members, at least one ring within the system being aromatic, and each ring within the system containing 3 to 12 ring members. Examples of "aryl" include, but are not limited to, phenyl, biphenyl, naphthyl, and anthracyl, etc., which may bear one or more substituents. As used herein, those also included within the scope of the term "aryl" are groups in which an aromatic ring is fused to one or more additional rings. In the case of a polycyclic ring system, only one of the rings need 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 include, but are not limited to, benzofuranyl, indanyl, phthalimidyl, naphthimidyl, phenanthridinyl, or tetrahydronaphthyl, etc. The aryl group may be substituted with substituents as described above at one or more ring positions.

[0025]

[0026] As used herein, the term "cyano" refers to -CN.

[0027] As used herein, the term "cycloalkyl", whether used as part of another term or independently, refers to a monovalent non-aromatic, saturated or partially unsaturated, monocyclic and polycyclic ring system, all of whose ring atoms are carbon and which contains at least 3 ring-forming carbon atoms. In some embodiments, the cycloalkyl may contain 3 to 12 ring-forming carbon atoms, 3 to 10 ring-forming carbon atoms, 3 to 9 ring-forming carbon atoms, 3 to 8 ring-forming carbon atoms, 3 to 7 ring-forming carbon atoms, 3 to 6 ring-forming carbon atoms, 3 to 5 ring-forming carbon atoms, 4 to 12 ring-forming carbon atoms, 4 to 10 ring-forming carbon atoms, 4 to 9 ring-forming carbon atoms, 4 to 8 ring-forming carbon atoms, 4 to 7 ring-forming carbon atoms, 4 to 6 ring-forming carbon atoms, 4 to 5 ring-forming 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 cyclic alkyl group containing at least one double bond or triple bond within its ring system. In some embodiments, the cycloalkyl group may be monocyclic or polycyclic. Condensed ring systems, spiro ring systems and bridged 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-cyclopenta-1-enyl, 1-cyclopenta-2-enyl, 1-cyclopenta-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, norbornyl, fluorenyl, spiro-pentadienyl, spiro[3.6]-decanyl, bicyclo[1,1,1]pentenyl, and bicyclo[2,2,1]heptenyl, etc.

[0026]

[0028] As used herein, the term "halogen" refers to an atom selected from fluorine (or fluoro), chlorine (or chloro), bromine (or bromo), and iodine (or iodo).

[0027]

[0029] As used herein, the term "heteroatom" refers to nitrogen, oxygen, or sulfur, and includes any oxidized form of nitrogen or sulfur, and any quaternized form of basic nitrogen (including N-oxides).

[0028]

[0030] As used herein, the term "heteroalkyl" refers to an alkyl in which at least one of the carbon atoms is replaced by a heteroatom selected from N, O, or S. Heteroalkyl may be a carbon radical or a heteroatom radical (i.e., the heteroatom may appear in the center or at the end of the radical), and may be independently optionally substituted with one or more substituents described herein. The term "heteroalkyl" includes alkoxy radicals and heteroalkoxy radicals.

[0029]

[0031] As used herein, the term "heteroalkenyl" refers to an alkenyl in which at least one of the carbon atoms is replaced by a heteroatom selected from N, O, or S. Heteroalkenyl may be a carbon radical or a heteroatom radical (i.e., the heteroatom may appear in the center or at the end of the radical), and may be independently optionally substituted with one or more substituents described herein.

[0030]

[0032] As used herein, the term "heteroalkynyl" refers to an alkynyl in which at least one of the carbon atoms is replaced by a heteroatom selected from N, O, or S. Heteroalkynyl may be a carbon radical or a heteroatom radical (i.e., the heteroatom may appear in the center or at the end of the radical), and may be independently optionally substituted with one or more substituents described herein.

[0031]

[0033] As used herein, the term "heteroaryl", whether used as part of another term or independently, refers to an aryl group having one or more heteroatoms in addition to carbon atoms. The heteroaryl group can be monocyclic. Examples of monocyclic heteroaryl include, but are not limited to, thienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl, purinyl, naphthyridinyl, benzofuranyl, and pteridinyl. The heteroaryl group also includes polycyclic groups in which the aromatic heterocycle is fused to one or more aryl rings, alicyclic rings or heterocyclyl rings, and the radical or point of attachment is on the aromatic heterocycle. Examples of polycyclic heteroaryl include, but are not limited to, indolyl, isoindolyl, benzothienyl, benzofuranyl, benzo[1,3]dioxolyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzthiazolyl, quinolyl, isoquinolyl, dihydroquinolinyl, dihydroisoquinolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H-quinolizinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, tetrahydroquinolinyl, and tetrahydroisoquinolinyl, etc.

[0032]

[0034] As used herein, the term "heterocyclyl" refers to a saturated or partially unsaturated carbocyclic group in which one or more ring atoms are heteroatoms independently selected from, for example, oxygen, sulfur, nitrogen, and phosphorus, and the remaining ring atoms are carbon, and one or more ring atoms may be independently optionally substituted with one or more substituents. In some embodiments, the heterocyclyl is a saturated heterocyclyl. In some embodiments, the heterocyclyl is a partially unsaturated heterocyclyl having one or more double bonds within its ring system. In some embodiments, the heterocyclyl may contain any oxidized form of carbon, nitrogen or sulfur, and any quaternized form of basic nitrogen. "Heterocyclyl" also includes radicals in which the heterocyclyl radical is fused to a saturated, partially unsaturated, or fully unsaturated (i.e., aromatic) carbocyclic or heterocyclic ring. The heterocyclyl radical may, where possible, be carbon-bonded or nitrogen-bonded. In some embodiments, the heterocyclic ring is carbon-bonded. In some embodiments, the heterocyclic ring is nitrogen-bonded. For example, a group derived from pyrrole may be pyrrol-1-yl (nitrogen-bonded) or pyrrol-3-yl (carbon-bonded). Further, a group derived from imidazole may be imidazol-1-yl (nitrogen-bonded) or imidazol-3-yl (carbon-bonded).

[0033]

[0035] In some embodiments, the term "3- to 12-membered heterocyclyl" refers to a 3- to 12-membered saturated or partially unsaturated monocyclic or polycyclic heterocyclic system having 1 to 3 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Condensed ring systems, spiro ring systems, and bridged ring systems are also included within the scope of this definition. Examples of monocyclic heterocyclyl include, but are not limited to, oxetanyl, 1,1-dioxothietanyl pyrrolidyl, tetrahydrofuryl, tetrahydrothienyl, pyrrolyl, furanyl, thienyl, pyrazolyl, imidazolyl, triazolyl, oxazolyl, thiazolyl, piperidyl, piperazinyl, piperidinyl, morpholinyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, triazinyl, pyridonyl, pyrimidonyl, pyrazinonyl, pyrimidonyl, pyridazonyl, pyrrolidinyl, and triazinonyl. Examples of condensed heterocyclyl include, but are not limited to, phenyl-condensed ring or pyridinyl-condensed ring, such as quinolinyl, isoquinolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, quinoxalinyl, quinolidinyl, quinazolinyl, azaindolizinyl, pteridinyl, chromenyl, isochromenyl, indolyl, isoindolyl, indolizinyl, indazolyl, purinyl, benzofuranyl, isobenzofuranyl, benzimidazolyl, benzothienyl, benzothiazolyl, carbazolyl, phenazinyl, phenothiazinyl, phenanthridinyl, hexahydro-1H-pyrrolidinyl, imidazo[1,2-a]pyridinyl, [1,2,4]triazolo[4,3-a]pyridinyl, and [1,2,3]triazolo[4,3-a]pyridinyl. Examples of spiro heterocyclyl include, but are not limited to, spiropyranyl, and spirooxazinyl. Examples of bridged heterocyclyl include, but are not limited to, morphanyl, hexamethylenetetraminyl, 3-aza-bicyclo[3.1.0]hexane, 8-aza-bicyclo[3.2.1]octane, 1-aza-bicyclo[2.2.2]octane, and 1,4-diazabicyclo[2.2.2]octane (DABCO).

[0034]

[0036] As used herein, the term "hydroxyl" or "hydroxy" refers to -OH.

[0037] As used herein, the term "partially unsaturated" refers to a radical containing at least one double bond or triple bond. The term "partially unsaturated" is intended to include rings having multiple sites of unsaturation, but not to include aromatic (i.e., fully unsaturated) moieties.

[0035]

[0038] As used herein, the terms "optional" or "optionally" mean that the subsequently described event or situation may or may not occur, and that the description includes both the case where the event or situation occurs and the case where it does not occur. As used herein, the term "substituted", whether or not preceded by the term "optionally", means that one or more hydrogens of the designated moiety are replaced by suitable substituents. Of course, "substitution" or "substituted with" implies that such substitution is in accordance with the allowable valence of the atom being substituted and that the substitution results in a stable or chemically feasible compound, e.g., the compound does not spontaneously undergo deformations such as by rearrangement, cyclization, elimination, etc. Unless otherwise specified, an "optionally substituted" group can have suitable substituents at each substitutable position of the group, and if two or more positions of any given structure can be substituted with two or more substituents selected from a particular group, the substituents can be the same or different at each position. Substituents include, but are not limited to, alkyl, alkenyl, alkynyl, alkoxy, acyl, amino, amide, amidino, aryl, azide, carbamoyl, carboxyl, carboxyl ester, cyano, guanidino, halo, haloalkyl, heteroalkyl, heteroaryl, heterocyclyl, hydroxy, hydrazino, imino, oxo, nitro, alkylsulfinyl, sulfonic acid, alkylsulfonyl, thiocyanate, thiol, thione, or combinations thereof. It will be understood by those skilled in the art that substituents themselves can be substituted where appropriate. Unless expressly stated to be "unsubstituted", references herein to chemical moieties are understood to include substituted isomers. For example, references to an "aryl" group or an "aryl" moiety implicitly include both substituted and unsubstituted isomers.

[0036]

[0039] As used herein, the term "substituted", whether preceded by the term "optionally" or not, means that one or more hydrogens of the designated moiety are replaced by an appropriate substituent. Of course, "substitution" or "substituted with" implies that such substitution is in accordance with the allowable valence of the atom being substituted and that the substitution results in a stable or chemically feasible compound, e.g., the compound does not undergo spontaneous transformations such as rearrangement, cyclization, elimination, etc. Unless otherwise specified, an "optionally substituted" group can have an appropriate substituent at each substitutable position of the group, and when two or more positions of any given structure can be substituted with two or more substituents selected from a particular group, the substituents can be the same or different at each position. It will be understood by those skilled in the art that substituents themselves can be substituted where appropriate. Unless expressly stated to be "unsubstituted", references to chemical moieties herein are understood to include substituted variants. For example, references to an "aryl" group or "aryl" moiety implicitly include both substituted and unsubstituted variants.

[0037]

[0040] The symbols "R" and "S" represent the configuration of substituents around a chiral carbon atom. The descriptors "R" and "S" for isomers are used herein as described and as defined in the literature (IUPAC Recommendations 1996, Pure and Applied Chemistry, 68:2193 - 2222 (1996)) to indicate the atomic configuration relative to the core molecule. Compound

[0041] In one aspect, the present disclosure provides a compound of formula (I):

[0038]

Chemical formula

[0039] [Wherein, R 1 and R2 Each of them is independently selected from the group consisting of hydrogen, hydroxy, halogen, cyano, amino, alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, and heteroalkynyl, and said alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, and heteroalkynyl are each optionally substituted with one or more groups independently selected from hydroxyl, halogen, cyano or amino, Each of X, W, Z, and U is independently N or C(R 3 ), R 3 is selected from the group consisting of hydrogen, hydroxy, halogen, cyano, amino, alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, and heteroalkynyl, and said alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, and heteroalkynyl are each optionally substituted with one or more groups independently selected from hydroxyl, halogen, cyano or amino, E is -N(R 4 )C(=O)-, -C(=O)N(R 4 )-, -N(R 4 )SO 2 -, or -SO 2 N(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 , wherein Y 1 is absent or -C(R 5 ) 2 , each R 5 is hydrogen, -N(R a ) 2, independently selected from the group consisting of alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, and -alkyl-heterocyclyl, and said alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl and -alkyl-heterocyclyl are one or more R 6 optionally substituted with each R 6 is 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 is independently selected from hydrogen or alkyl, Y 2 is -(CH 2 ) p -cycloalkyl-*, -(CH 2 ) p -heterocyclyl-*, -(CH 2 ) p -aryl-*, or -(CH 2 ) p -heteroaryl-*, each of which may be optionally substituted with one or more groups independently selected from hydroxy, halogen, cyano, amino, alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, or heteroalkynyl, p is 0, 1 or 2, and the * end of Y 2 is attached to Y 3 Y 3 is absent or -Y 31 -Y 32 -Y 33 wherein, Y 31 is absent or selected from alkyl, alkenyl or alkynyl, Y 32 ​​is 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 )- and is selected from each R b is independently selected from hydrogen or alkyl, and the #-end of Y 32 is bonded to Y 33 and Y 33 is 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, and the alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl are optionally substituted with one or more R 7 and each R 7 is independently selected from halogen, hydroxy, amino, cyano, nitrooxy, alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, or haloalkyl, each R c is independently selected from hydrogen or alkyl, and n is an integer from 1 to 5]] and provides a compound of or a pharmaceutically acceptable salt thereof.

[0040]

[0042] In some embodiments, each of R 1 and R 2 is independently hydrogen, halogen, hydrogen, cyano, amino, or alkyl optionally substituted with one or more halogens.

[0041]

[0043] In some embodiments, R 1 is hydrogen or alkyl, and R 2is hydrogen, halogen, or alkyl. In certain embodiments, R 1 is hydrogen or C 1~6 alkyl, and R 2 is hydrogen, halogen, or C 1~6 alkyl.

[0042]

[0044] In some embodiments, R 1 and R 2 are both hydrogen.

[0045] In some embodiments, R 1 and R 2 are both alkyl. In certain embodiments, R 1 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 is C 1~6 alkyl, C 1~5 alkyl, C 1~4 alkyl, C 1~3 alkyl, or C 1~2 alkyl. In certain embodiments, R 1 is methyl and R 2 is methyl.

[0043]

[0046] In some embodiments, R 1 is hydrogen and R 2 is halogen. In certain embodiments, R 1 is hydrogen and R 2 is fluoro.

[0047] In some embodiments, R 1 is hydrogen and R 2 is alkyl. In certain embodiments, R 1 is hydrogen and R 2 is C 1~6 alkyl, C 1~5 alkyl, C 1~4 alkyl, C 1~3 alkyl, or C 1~2 alkyl. In certain embodiments, R1 is hydrogen, and R 2 is methyl.

[0044]

[0048] In some embodiments, X, W, U, and Z are C(R 3 ). In certain embodiments, W, U, and Z are C(R 3 ), and R 3 is hydrogen.

[0049] In some embodiments, X and U are C(R 3 ), and W and Z are N. In certain embodiments, X and U are C(R 3 ), R 3 is hydrogen, and W and Z are N.

[0045]

[0050] In some embodiments, E is -N(R 4 )C(=O)- or -C(=O)N(R 4 ). In certain embodiments, E is -N(R 4 )C(=O)- or -C(=O)N(R 4 ), and R 4 is hydrogen.

[0046]

[0051] In some embodiments, Y 1 is absent.

[0052] In some embodiments, Y 1 is -C(R 5 ) 2 .

[0053] In certain embodiments, Y 1 is -C(R 5 ) 2 , one of R 5 is hydrogen, and the other R 5 is selected from -N(R a ), alkyl, cycloalkyl, heterocyclyl, and -alkyl-heterocyclyl, and the alkyl, cycloalkyl, heterocyclyl, and -alkyl-heterocyclyl are optionally substituted with one or more R 2 6 . ​

[0047]

[0054] In certain embodiments, Y 1 is -C(R 5 ) 2 -, where one R 5 is hydrogen and the other R 5 is selected from -N(R a ) 2 , C 1~6 alkyl, C 3~12 cycloalkyl, 5- to 12-membered heterocyclyl, or -(C 1~6 alkyl)-(5- to 12-membered heterocyclyl), and the C 1~6 alkyl, C 3~12 cycloalkyl, 5- to 12-membered heterocyclyl, and -(C 1~6 alkyl)-(5- to 12-membered heterocyclyl) are optionally substituted with one or more R 6 .

[0048]

[0055] In certain embodiments, Y 1 is -C(R 5 ) 2 -, where one R 5 is alkyl and the other R 5 is selected from -N(R a ) 2 , alkyl, cycloalkyl, or heterocyclyl, and the alkyl, cycloalkyl, and heterocyclyl are optionally substituted with one or more R 6 .

[0049]

[0056] In certain embodiments, Y 1 is -C(R 5 ) 2 -, where one R 5 is C 1~6 alkyl and the other R 5 is selected from -N(R a ) 2 , C 1~6 alkyl, C 3~12 cycloalkyl, or 5- to 12-membered heterocyclyl, and the C 1~6 alkyl, C 3~12The cycloalkyl or 5- to 12-membered heterocyclyl is optionally substituted with one or more Rs 6 and is optionally substituted with one or more Rs

[0050]

[0057] In some embodiments, R 6 is selected from amino, alkyl, or -alkyl-N(R a ) 2 . In certain embodiments, R 6 is selected from amino, C 1~6 alkyl, or -(C 1~6 alkyl)-N(R a ) 2 .

[0051]

[0058] In some embodiments, Y 1 is

[0052]

Chemical formula

[0053] selected from the group consisting of

[0059] In some embodiments, Y 2 is -(CH 2 ) p -cycloalkyl-*.

[0060] In certain embodiments, Y 2 is -(CH 2 ) p -(C 3~10 cycloalkyl)-*, and p is 0 or 1.

[0054]

[0061] In certain embodiments, Y 2 is cyclopentyl or cyclohexyl.

[0062] In certain embodiments, Y 2 is -(CH 2 ) p -heterocyclyl-*.

[0063] In certain embodiments, Y 2 is -(CH 2 )p -(heterocyclyl of 5 to 12 members)-*, and p is 0 or 1.

[0055]

[0064] In certain embodiments, Y 2 is tetrahydrofuranyl, pyrrolidinyl, or tetrahydropyranyl.

[0065] In some embodiments, Y 2 is -(CH 2 ) p -aryl-*.

[0056]

[0066] In certain embodiments, Y 2 is -(CH 2 ) p -(C 5~12 aryl)-*, and p is 0 or 1.

[0067] In certain embodiments, Y 2 is phenyl or -CH 2 -phenyl-*.

[0057]

[0068] In some embodiments, Y 2 is -(CH 2 ) p -heteroaryl-*.

[0069] In certain embodiments, Y 2 is -(CH 2 ) p -(heteroaryl of 5 to 12 members)-*, and p is 0 or 1.

[0058]

[0070] In certain embodiments, Y 2 is pyridinyl, tetrahydroisoquinolinyl, -CH 2 -imidazolyl-*, or -CH 2 -indolyl-*.

[0071] In some embodiments, Y 31 is absent or alkyl. In certain embodiments, Y 31 is absent or C 1~6 alkyl, C 1~5 alkyl, C1~4 alkyl, C 1~3 alkyl, or C 1~2 is alkyl.

[0059]

[0072] In some embodiments, Y 31 is absent, and Y 32 is absent or is selected from -O-#, -C(=O)O-#, -P(=O)(R b )-#, -N(R b )C(=O)-#, or -C(=O)N(R b )-#.

[0060]

[0073] In some embodiments, Y 31 is absent, and Y 32 is absent or is selected from -O-#, -C(=O)O-#, -P(=O)(R b )-#, -N(R b )C(=O)-#, or -C(=O)N(R b )-#, and Y 33 is selected from -N(R c ) 2 , alkyl, or aryl, wherein the alkyl and aryl are optionally substituted with one or more R 7 .

[0061]

[0074] In some embodiments, Y 31 is absent, and Y 32 is absent or is selected from -O-#, -C(=O)O-#, -P(=O)(R b )-#, -N(R b )C(=O)-#, or -C(=O)N(R b )-#, and Y 33 is 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 are optionally substituted with one or more R 7 .

[0062]

[0075] In certain embodiments, Y 31 is absent, and Y 32 is either absent or selected from -O-#, -C(=O)O-#, -P(=O)(R b )-#, -N(R b )C(=O)-#, or -C(=O)N(R b )-#, and Y 33 is -N(R c ) 2 , C 1~6 alkyl, or C 5~12 aryl, where the C 1~6 alkyl and C 5~12 aryl are independently optionally substituted with one or more R 7 selected from halogen, hydroxy, amino, cyano, nitrooxy, or alkyl.

[0063]

[0076] In certain embodiments, Y 31 is absent, and Y 32 is either absent or selected from -O-#, -C(=O)O-#, -P(=O)(R b )-#, -N(R b )C(=O)-#, or -C(=O)N(R b )-#, and Y 33 is selected from hydrogen, hydroxyl, cyano, halogen, -NH 2 , methyl, -CH 2 CH 2 OCH 3 .

[0064]

[0077] In some embodiments, Y 31 is alkyl, and Y 32 is either absent or selected from -O-#, -OC(=O)-#, -OC(=O)N(R b )-#, or -N(R b )C(=O)-#.

[0065]

[0078] In some embodiments, Y 31 is C 1~6 alkyl, C 1~5 alkyl, C 1~4 alkyl, C1~3 Alkyl, or C 1~2 alkyl, and Y 32 is either absent, -O-#, -OC(=O)-#, -OC(=O)N(R b )-#, or -N(R b )C(=O)-#.

[0066]

[0079] In some embodiments, Y 31 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 is either absent, -O-#, -OC(=O)-#, -OC(=O)N(R b )-#, or -N(R b )C(=O)-#, and Y 33 is -N(R c ) 2 , alkyl, or aryl, where the alkyl and aryl are optionally substituted with one or more R 7 s.

[0067]

[0080] In some embodiments, Y 31 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 is either absent, -O-#, -OC(=O)-#, -OC(=O)N(R b )-#, or -N(R b )C(=O)-#, and Y 33 is -NH 2 , -N(CH 3 ) 2 , methyl, dimethylphenyl, or nitrooxypentyl, where the alkyl and aryl are optionally substituted with one or more R 7 s.

[0068]

[0081] In certain embodiments, Y 31 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 is absent or selected from -O-#, -OC(=O)-#, -OC(=O)N(R b ), -# or -N(R b ),C(=O)-#, and Y 33 is -NH 2 , -N(CH 3 ), 2 methyl, dimethylphenyl, or nitrooxypentyl.

[0069]

[0082] In some embodiments, n is 1 or 2.

[0083] Exemplary compounds of formula (I) are shown in Table 1 below.

[0070]

Table 1-1

[0071]

Table 1-2

[0072]

Table 1-3

[0073]

Table 1-4

[0074]

Table 1-5

[0075]

Table 1-6

[0076]

Table 1-7

[0077]

Table 1-8

[0078]

Table 1-9

[0079]

Table 1-10

[0080]

Table 1-11

[0081]

Table 1-12

[0082]

Table 1-13

[0083]

Table 1-14

[0084]

Table 1-15

[0085]

[0084] The compounds provided in this specification are described with respect to both general formulas and specific compounds. In addition, the compounds of the present disclosure may exist in a number of different forms or derivatives, including but not limited to prodrugs, active metabolite derivatives (active metabolic products), solvates, pharmaceutically acceptable salts, or isotope derivatives, all of which are within the scope of the present disclosure.

[0086] As used herein, the term "prodrug" refers to a compound or a pharmaceutically acceptable salt thereof that yields the desired active compound when metabolized under physiological conditions or when converted by solvolysis. Prodrugs include, without limitation, esters, amides, carbamates, carbonates, ureas, solvates, or hydrates of the active compound. Usually, prodrugs are inactive or less active than the active compound, but can provide one or more advantageous handling, administration, and / or metabolic properties. For example, some prodrugs are esters of the active compound and the ester group is cleaved during metabolic degradation to yield the active drug. Also, some prodrugs yield a compound that is enzymatically activated to yield the active compound or yields a compound that undergoes a further chemical reaction to yield the active compound. A prodrug can proceed from the prodrug form to the active form in a single step or can have one or more intermediate forms that can themselves be active or inactive. The preparation and use of prodrugs are discussed in T. Higuchi and V. Stella, "Pro-drugs as Novel Delivery Systems", Vol. 14 of the A.C.S. Symposium Series; Bioreversible Carriers in Drug Design, edited by Edward B. Roche, American Pharmaceutical Association and Pergamon Press, 1987; Prodrugs: Challenges and Rewards, edited by V. Stella, R. Borchardt, M. Hageman, R. Oliyai, H. Maag, J. Tilley, Springer-Verlag New York, 2007, all of which are hereby incorporated by reference in their entirety.

[0087] As used herein, the term "metabolite", such as an active metabolite, has something in common with the above prodrug. Thus, such a metabolite is a pharmacologically active compound, or a compound that further metabolizes into a pharmacologically active compound that is a derivative produced by the metabolic process in the body of the subject. For example, such metabolites may be produced by oxidation, reduction, hydrolysis, amidation, amidolysis, esterification, esterolysis, and enzymatic cleavage of the administered compound or salt or prodrug. Among these, the active metabolite is such a pharmacologically active derivative compound. For prodrugs, the prodrug compound is generally inactive or less active than the metabolite. For active metabolites, the parent compound may be an active compound or an inactive prodrug.

[0088]

[0087] Prodrugs and active metabolites can be identified using conventional techniques known in the art. See, for example, Bertolini et al., 1997, J Med Chem 40, pp. 2011-2016; Shan et al., J Pharm Sci 86, pp. 756-757; Bagshawe, 1995, DrugDev Res 34, pp. 220-230; Wermuth, supra.

[0089]

[0088] As used herein, the term "pharmaceutically acceptable" indicates that a substance or composition is chemically and / or toxicologically compatible with the other components that make up the formulation and / or the subject being treated with the formulation.

[0090] As used herein, the term "pharmaceutically acceptable salt" includes salts that maintain the biological effects of the free acids and free bases of a particular compound and are not biologically or otherwise harmful, unless otherwise specified. The intended pharmaceutically acceptable salt forms include, but are not limited to, mono, bis, tris, and tetrakis, etc. Pharmaceutically acceptable salts are non-toxic at the amounts and concentrations at which they are administered. When such salts are prepared, the pharmacological use can be facilitated by changing the physical properties of the compound without preventing the compound from exerting its physiological effects. Useful changes in physical properties include lowering the melting point to facilitate transmucosal administration and increasing the solubility to facilitate administration at higher concentrations of the drug.

[0091]

[0090] Pharmaceutically acceptable salts include acid addition salts, such as acid addition salts containing sulfate ion, chloride ion, hydrochloride ion, fumarate ion, maleate ion, phosphate ion, sulfamate ion, acetate ion, citrate ion, lactate ion, tartrate ion, methanesulfonate ion, ethanesulfonate ion, benzenesulfonate ion, p-toluenesulfonate ion, cyclohexylsulfamate ion, and quinate ion. Pharmaceutically acceptable salts can be obtained from acids such as hydrochloric acid, maleic acid, sulfuric acid, phosphoric acid, sulfamic acid, acetic acid, citric acid, lactic acid, tartaric acid, malonic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, cyclohexylsulfamic acid, fumaric acid, and quinic acid.

[0092]

[0091] Pharmaceutically acceptable salts include base addition salts when acidic functional groups such as carboxylic acids or phenols are present, for example, salts containing benzathine, chloroprocaine, choline, diethanolamine, ethanolamine, t-butylamine, ethylenediamine, meglumine, procaine, aluminum, calcium, lithium, magnesium, potassium, sodium, ammonium, alkylamine, and zinc. See, for example, Remington’s Pharmaceutical Sciences, 19th Edition, Mack Publishing Co., Easton, PA, Volume 2, page 1457, 1995; “Handbook of Pharmaceutical Salts: Properties, Selection, and Use” by Stahl and Wermuth, Wiley-VCH, Weinheim, Germany, 2002. Such salts can be prepared using an appropriate corresponding base.

[0093]

[0092] Pharmaceutically acceptable salts can be prepared by standard techniques. For example, a compound in free base form can be dissolved in an appropriate solvent such as an aqueous solution or aqueous alcohol solution containing an appropriate acid, and then the solution can be isolated by evaporation. Thus, when a particular compound is a base, the desired pharmaceutically acceptable salt can be prepared by any appropriate method available in the art, for example, using an inorganic acid such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, and phosphoric acid, or an organic acid such as acetic acid, maleic acid, succinic acid, mandelic acid, fumaric acid, malonic acid, pyruvic acid, oxalic acid, glycolic acid, salicylic acid, pyranosidic acid (such as glucuronic acid or galacturonic acid), alpha-hydroxy acid (such as citric acid or tartaric acid), amino acid (such as aspartic acid or glutamic acid), aromatic acid (such as benzoic acid or cinnamic acid), or sulfonic acid (such as p-toluenesulfonic acid or ethanesulfonic acid) to treat the free base.

[0094]

[0093] Similarly, when the specific compound is an acid, the desired pharmaceutically acceptable salt may be prepared by any suitable method, for example, by treating the free acid with an inorganic base or an organic base, such as an amine (primary, secondary or tertiary), or an alkali metal hydroxide or an alkaline earth metal hydroxide. Examples useful in the description of suitable salts include organic salts derived from amino acids (e.g., L-glycine, L-lysine, and L-arginine), ammonia, primary, secondary, and tertiary amines, and cyclic amines (e.g., hydroxyethylpyrrolidine, piperidine, morpholine or piperazine), and inorganic salts derived from sodium, calcium, potassium, magnesium, manganese, iron, copper, zinc, aluminum and lithium.

[0095]

[0094] Naturally, the compounds of the present disclosure can exist in unsolvated form, solvated form (e.g., hydrated form), and solid form (e.g., crystalline or polymorphic form), and the present disclosure is intended to embrace all such forms.

[0096]

[0095] As used herein, the terms "solvate" or "solvated form" refer to a solvate addition form containing a solvent in either a stoichiometric amount or a non-stoichiometric amount. Some compounds tend to capture solvent molecules in a fixed molar ratio in the crystalline solid state, thereby forming a solvate. When the solvent is water, the solvate formed is a hydrate, and when the solvent is an alcohol, the solvate formed is an alcoholate. Hydrates are formed by the combination of one or more molecules of water with one molecule of a substance, where the water retains its molecular state as H 2 O. Examples of solvents that form solvates include, but are not limited to, water, isopropanol, ethanol, methanol, DMSO, ethyl acetate, acetic acid, and ethanolamine.

[0097]

[0096] This disclosure is to include all isotopes of atoms in the compounds. Isotopes of an atom include atoms that have the same atomic number but different mass numbers. For example, unless otherwise specified, hydrogen, carbon, nitrogen, oxygen, phosphorous, sulfur, fluorine, chlorine, bromide, or iodine in the compounds of this disclosure are their isotopes, such as, but not limited to, 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 are also to be included. In some embodiments, hydrogen includes protium, deuterium, and tritium. In some embodiments, carbon is 12 C and 13 C.

[0098]

[0097] The compounds or pharmaceutically acceptable salts thereof provided herein can contain one or more asymmetric centers and, thus, can be defined as enantiomers, diastereomers, and other stereoisomeric forms as (R)- or (S)- from the perspective of absolute stereochemistry, or as (D)- or (L)- for amino acids, and can give rise to other stereoisomeric forms that can be defined as rel-(R)- or rel-(S)- from the perspective of relative configuration. The present disclosure includes all such possible isomers, as well as their racemic and optically pure forms. Optically active (+) and (−) isomers, (R)- and (S)-isomers, or (D)- and (L)-isomers can be prepared using chiral synthons or chiral reagents or can be separated by conventional techniques such as chromatography and fractional crystallization. Conventional techniques for the preparation and isolation of individual enantiomers include chiral synthesis from optically pure and suitable precursors or resolution of a racemate (or a racemate of a salt or derivative) using, for example, chiral high performance liquid chromatography (HPLC). When a compound is represented in its chiral form, it will of course be understood that its embodiments include, but are not limited to, specific diastereomeric or enantiomer-enriched forms. In situations where chirality is present but not specified, it will of course be understood that its embodiments include either specific diastereomeric or enantiomer-enriched forms, or a racemic mixture of such compounds or a scalemic mixture of enantiomers.

[0099]

[0098] The term “stereoisomer” refers to compounds that contain the same atoms bonded by the same bonds but have different three-dimensional structures and cannot interconvert. The present disclosure contemplates various stereoisomers and mixtures thereof and includes “enantiomers,” which refer to two stereoisomers that are mirror images of each other and cannot be superimposed on each other.

[0100] As used herein, the term "enantiomer" refers to a pair of stereoisomers that are mirror images of each other and cannot be superimposed. A 1:1 mixture of a pair of enantiomers is a "racemic" mixture. A mixture in a ratio other than 1:1 of enantiomers is a "scalemic" mixture of enantiomers.

[0101] As used herein, the term "diastereoisomer" refers to stereoisomers that have at least two asymmetric atoms but are not mirror images of each other. As used herein, the term "tautomer" or "tautomeric form" refers to structurally isomeric forms of a compound that are interconvertible with a low energy barrier. The existence and concentration of these tautomeric forms will depend on the environment in which the compound is found and may vary, for example, depending on whether the compound is a solid, an organic solution, or an aqueous solution. By way of example, proton tautomers (also known as prototropic tautomers) include interconversions via the movement of a proton, such as the isomerization of keto-enol, amide-imido acid, lactam-lactim, imine-enamine, and cyclic forms in which a proton can occupy two or more positions in a heterocyclic system. Valence tautomers include interconversions by the rearrangement of some of the bonding electrons. Tautomers can be in an equilibrium state or can be stereochemically fixed in one form by appropriate substitution. Compounds of the present disclosure identified as a particular tautomeric form by name or structure are to be considered to include other tautomeric forms unless otherwise specified.

[0102] When the compounds provided herein contain an olefinic double bond or other geometrically asymmetric center and unless otherwise specified, these compounds include both E and Z geometric isomers.

[0103] Synthetic Methods The compounds provided herein can be prepared using any known organic synthesis technique and can be synthesized according to any of a number of available synthetic routes.

[0104]

[0104] The reactions for preparing the compounds of the present disclosure can be carried out in a suitable solvent that can be readily selected by one of ordinary skill in organic synthesis. A suitable solvent is substantially non-reactive with the starting materials (reactants), intermediates, and products at the temperature at which the reaction is carried out, which can be, for example, a temperature in the range from the freezing temperature of the solvent to the boiling temperature of the solvent. A given reaction can be carried out in one solvent or in a mixture of more than one solvent. Depending on the particular reaction step, a suitable solvent for the particular reaction step can be selected by one of ordinary skill in the art.

[0105]

[0105] The preparation of the compounds of the present disclosure may require the protection and deprotection of various chemical groups. The need for protection and deprotection and the selection of suitable protecting groups can be readily determined by one of ordinary skill in the art. 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 Edition, 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 Edition, Wiley, 2014, all of which are hereby incorporated by reference in their entirety.

[0106]

[0106] The reaction can be monitored according to any suitable method known in the art. For example, the formation of the product can be monitored by spectroscopic means, such as nuclear magnetic resonance spectroscopy (e.g., 1 H or 13C), can be monitored by infrared spectroscopy, spectrophotometry (e.g., ultraviolet-visible), mass spectrometry, or by chromatography methods such as high performance liquid chromatography (HPLC), liquid chromatography-mass spectrometry (LCMS), or thin layer chromatography (TLC). Compounds can be purified by those skilled in the art by various 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), pp. 874-883, which is incorporated herein by reference in its entirety), and normal phase silica chromatography.

[0107] Pharmaceutical composition

[0107] In a further aspect, there is provided a pharmaceutical composition comprising one or more compounds of the present disclosure or a pharmaceutically acceptable salt thereof. In some embodiments, the pharmaceutical composition of the present disclosure comprises a first compound of formula (I) or a pharmaceutically acceptable salt thereof, and one or more additional compounds of the same formula, provided that the first compound and the additional compounds are not the same molecule.

[0108]

[0108] In another aspect, there is provided a pharmaceutical composition comprising one or more compounds of the present disclosure or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable excipient.

[0109]

[0109] In some embodiments, the pharmaceutical composition of the present disclosure comprises a therapeutically effective amount of one or more compounds of the present disclosure or a pharmaceutically acceptable salt thereof.

[0110] In some embodiments, the pharmaceutical composition of the present disclosure comprises a therapeutically effective amount of one or more compounds of the present disclosure or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable excipient.

[0110]

[0111] As used herein, the term "therapeutically effective amount" refers to an amount of a molecule, compound, or composition comprising a molecule or compound for treating, ameliorating, or preventing a specified disease or condition, or for exhibiting a detectable therapeutic or inhibitory effect. The effect can be detected by any assay known in the art. The exact effective amount for a subject will depend upon the subject's weight, size, and health, the nature and extent of the condition, the rate of administration, the therapeutic agent or combination of therapeutic agents selected for administration, and the judgment of the prescribing physician. The therapeutically effective amount in a particular situation can be determined by routine experimentation within the skill and judgment of the clinician.

[0111]

[0112] As used herein, the term "pharmaceutical composition" refers to a formulation containing the molecules or compounds of the present disclosure in a form suitable for administration to a subject. Pharmaceutical compositions include suitable compositions adapted for oral, rectal, topical, parenteral (including subcutaneous, intramuscular, and intravenous), sublingual, ophthalmic, transdermal, or nasal administration, but in each case the optimal route will depend on the particular host, as well as the nature and severity of the condition to which the active ingredient is being administered. Pharmaceutical compositions can be conveniently provided in unit dosage form and can be prepared by any of the methods well known in the art of pharmacy.

[0112]

[0113] As used herein, the term "pharmaceutically acceptable excipient" means an excipient that is generally safe, non-toxic, and neither biologically nor otherwise harmful, and that is useful in the preparation of pharmaceutical compositions, including excipients acceptable for veterinary use and for human pharmaceutical use. "Pharmaceutically acceptable excipient" as used herein includes both a single such excipient and more than one such excipient. The term "pharmaceutically acceptable excipient" also encompasses "pharmaceutically acceptable carrier" and "pharmaceutically acceptable diluent".

[0113]

[0114] The specific excipients used will depend on the means and purposes to which the compounds of the present disclosure are applied. Solvents are generally selected based on solvents recognized by those skilled in the art as being safe for administration to mammals, including humans. Generally, safe solvents are non-toxic aqueous solvents such as water, and other non-toxic solvents that are soluble or miscible in water. Suitable aqueous solvents include water, ethanol, propylene glycol, polyethylene glycols (e.g., PEG400, PEG300), and mixtures thereof.

[0114]

[0115] In some embodiments, suitable excipients include buffering agents such as phosphate buffers, citrate buffers, and other organic acid buffers; antioxidants such as ascorbic acid and methionine; preservatives (octadecyl dimethyl benzyl ammonium chloride, hexamethonium chloride, benzalkonium chloride, benzethonium chloride, phenolic alcohol, butyl alcohol or benzyl alcohol, alkyl parabens such as methyl paraben or propyl paraben, catechol, resorcinol, cyclohexanol, 3-pentanol, and m-cresol); low molecular weight (less than about 10 residues) polypeptides; proteins such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinyl pyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides and other carbohydrates such as 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., Zn-protein complexes); and / or non-ionic surfactants such as TWEEN™, PLURONICS™ or polyethylene glycol (PEG).

[0115]

[0116] In some embodiments, suitable excipients can include one or more stabilizers, surfactants, wetting agents, lubricants, emulsifiers, suspending agents, preservatives, antioxidants, opacifying agents, flow promoters, processing aids, colorants, sweeteners, fragrances, flavoring agents, and other known additives to provide a fine appearance to the drug (i.e., the compounds or pharmaceutical compositions of the present disclosure) or to assist in the manufacture of a pharmaceutical product (i.e., a medicine). The active pharmaceutical ingredient can also be encapsulated, for example, in microcapsules prepared by coacervation techniques or interfacial polymerization, such as in microcapsules of hydroxymethylcellulose or gelatin, respectively, and poly-(methyl methacrylate) microcapsules, in colloidal drug delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles, and nanocapsules), or in macroemulsions. Such techniques are disclosed in Remington’s Pharmaceutical Sciences, 16th Edition, Osol, A. Ed. (1980). “Liposomes” are small vesicles composed of various types of lipids, phospholipids, and / or surfactants that are useful for the delivery of drugs (compounds disclosed herein, and optionally chemotherapeutic agents, etc.) to mammals, including humans. The components of liposomes are generally arranged in a bilayer structure similar to the lipid arrangement of biological membranes.

[0116]

[0117] The pharmaceutical compositions provided herein can be in any form that enables the composition to be administered to a subject, including but not limited to humans, and can be formulated to be compatible with the intended route of administration.

[0117]

[0118] Various routes are contemplated for the pharmaceutical compositions provided herein, and thus, the pharmaceutical compositions provided herein can be supplied in bulk or in unit dosage form, depending on the intended route of administration. For example, for oral, buccal, and sublingual administration, powders, suspensions, granules, tablets, pills, capsules, gel caps, and caplets may be acceptable as solid dosage forms, and emulsions, syrups, elixirs, suspensions, and solutions may be acceptable as liquid dosage forms. For parenteral administration, emulsions and suspensions may be acceptable as liquid dosage forms, and powders suitable for reconstitution with an appropriate solution may be acceptable as solid dosage forms. For inhalation administration, solutions, sprays, dry powders, and aerosols may be acceptable dosage forms. For topical (including buccal and sublingual) or transdermal administration, powders, sprays, ointments, pastes, creams, lotions, gels, solutions, and patches may be acceptable dosage forms. For vaginal administration, vaginal suppositories, tampons, creams, gels, pastes, foams, and sprays may be acceptable dosage forms.

[0118]

[0119] In some embodiments, the pharmaceutical compositions of the disclosure can be in the form of a formulation for oral administration.

[0120] In certain embodiments, the pharmaceutical compositions of the present disclosure can be in the form of tablet formulations. Suitable pharmaceutically acceptable excipients for tablet formulations include, for example, inert diluents such as lactose, sodium carbonate, calcium phosphate or calcium carbonate, granulating and disintegrating agents such as corn starch or alginic acid; binders such as starch; lubricants such as magnesium stearate, stearic acid or talc; preservatives such as ethyl p-hydroxybenzoate or propyl p-hydroxybenzoate, and antioxidants such as ascorbic acid. The tablet formulations may or may not be coated, but if coated, it is to modify the disintegration of the formulation and subsequent absorption of the active ingredient in the gastrointestinal tract, or to improve the stability and / or appearance of the formulation, and in either case, conventional coating agents and procedures well known in the art are used.

[0119]

[0121] In certain embodiments, the pharmaceutical compositions of the present disclosure may be in the form of hard gelatin capsules in which the active ingredient is mixed with an inert solid diluent such as calcium carbonate, calcium phosphate or kaolin, or may be in the form of soft gelatin capsules in which the active ingredient is mixed with water or an oil such as peanut oil, liquid paraffin or olive oil.

[0120]

[0122] In certain embodiments, the pharmaceutical composition of the present disclosure can be in the form of an aqueous suspension, which generally contains the active ingredient in fine powder form together with one or more suspending agents such as sodium carboxymethyl cellulose, methyl cellulose, hydroxypropylmethyl cellulose, sodium alginate, polyvinyl-pyrrolidone, tragacanth gum and gum arabic, dispersing agents or wetting agents such as lecithin, or condensation products of alkylene oxides and fatty acids (such as polyoxethylene stearate), or condensation products of ethylene oxide and long-chain aliphatic alcohols such as heptadecaethyleneoxycetanol, or condensation products of ethylene oxide and partial esters derived from fatty acids and hexitols such as polyoxyethylene sorbitol monooleate, or condensation products of ethylene oxide and partial esters derived from fatty acids and hexitol anhydrides such as polyethylene sorbitan monooleate. The aqueous suspension may also contain one or more preservatives (such as ethyl p-hydroxybenzoate or propyl p-hydroxybenzoate), antioxidants (such as ascorbic acid), coloring agents, flavoring agents, and / or sweetening agents (such as sucrose, saccharin or aspartame).

[0121]

[0123] In certain embodiments, the pharmaceutical composition of the present disclosure can be in the form of an oily suspension, which generally contains the active ingredient suspended in a vegetable oil (such as castor oil, olive oil, sesame oil or coconut oil) or in a mineral oil (such as liquid paraffin). The oily suspension may also contain thickening agents such as beeswax, solid paraffin or cetyl alcohol. Sweetening agents and flavoring agents such as those described above may be added to provide a palatable oral formulation. These compositions can be preserved by the addition of antioxidants such as ascorbic acid.

[0122]

[0124] In certain embodiments, the pharmaceutical compositions of the present disclosure can be in the form of oil-in-water emulsions. The oily phase can be a vegetable oil such as olive oil or peanut oil, or a mineral oil such as liquid paraffin, or a mixture of any of these. Suitable emulsifiers can be, for example, natural gums such as gum arabic or tragacanth gum, natural phospholipids such as soybean lecithin, esters or partial esters derived from fatty acids and hexitol anhydrides (e.g., sorbitan monooleate), and condensation products of said partial esters with ethylene oxide, such as polyoxyethylene sorbitan monooleate. The emulsion may also contain sweeteners, flavoring agents and preservatives.

[0123]

[0125] In certain embodiments, the pharmaceutical compositions provided herein can be in the form of syrups and elixirs, which may contain sweeteners such as glycerol, propylene glycol, sorbitol, aspartame or sucrose, demulcents, preservatives, flavoring agents and / or coloring agents.

[0124]

[0126] In some embodiments, the pharmaceutical compositions of the present disclosure can be in the form of formulations for parenteral administration.

[0127] In certain embodiments, the pharmaceutical compositions of the present disclosure can be in the form of sterile injectable formulations, such as sterile aqueous suspensions or oily suspensions for injection. This suspension can be formulated according to known techniques using the appropriate dispersing or wetting agents and suspending agents described above. The sterile injectable formulation can also be a sterile injectable solution or a sterile injectable suspension in a non-toxic parenterally acceptable diluent or solvent, such as a solution in 1,3-butanediol, and may be prepared as a lyophilized powder. Acceptable vehicles and solvents that may be used include water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile fixed oils can conveniently be used as a solvent or suspending medium. For this purpose, any bland fixed oil containing synthetic monoglycerides or diglycerides may be used. In addition, fatty acids such as oleic acid may likewise be used in the preparation of injectables.

[0125]

[0128] In some embodiments, the pharmaceutical composition of the present disclosure can be in the form of a formulation for inhalation administration.

[0129] In certain embodiments, the pharmaceutical composition of the present disclosure can be in the form of an aqueous and non-aqueous (e.g., in a fluorocarbon propellant) aerosol containing any suitable solvent and optionally other compounds such as, but not limited to, stabilizers, antimicrobial agents, antioxidants, pH adjusters, surfactants, bioavailability modifiers, and combinations thereof. Carriers and stabilizers vary with the requirements of the particular compound, but typically include nonionic surfactants (Tween, Pluronic, or polyethylene glycol), innocuous proteins such as serum albumin, sorbitan esters, oleic acid, lecithin, amino acids such as glycine, buffers, salts, sugars, or sugar alcohols.

[0126]

[0130] In some embodiments, the pharmaceutical composition of the present disclosure can be in the form of a formulation for topical or transdermal administration. In certain embodiments, the pharmaceutical compositions provided herein can be in the form of creams, ointments, gels, and aqueous or oily solutions or suspensions, which generally formulate the active ingredient with excipients acceptable for conventional topical use such as, for example, animal and vegetable fats, oils, waxes, paraffins, starches, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonite, silicic acid, talc, and zinc oxide, or mixtures thereof.

[0127]

[0131] In certain embodiments, the pharmaceutical compositions provided herein may be formulated for ophthalmic administration. In certain embodiments, the pharmaceutical compositions provided herein can be in the form of ophthalmic formulations such as, for example, ophthalmic ointments, ophthalmic powders, and ophthalmic solutions. In certain embodiments, the ophthalmic formulations are prepared at a comfortable pH using a suitable buffer system.

[0128]

[0132] In addition to the representative dosage forms described above, pharmaceutically acceptable excipients and carriers are generally known to those of ordinary skill in the art and are thus included in the present disclosure. Such excipients and carriers are described, for example, in "Remingtons Pharmaceutical Sciences", Mack Pub. Co., New Jersey (1991), "Remington: The Science and Practice of Pharmacy", edited by University of the Sciences in Philadelphia, 21st Edition, LWW (2005), which are hereby incorporated by reference into this specification.

[0129]

[0133] The dosing regimens provided herein will vary depending on known factors such as the pharmacodynamic properties of the particular agent and its mode and route of administration; the species, age, sex, health status, medical condition, and weight of the recipient; the nature and extent of the symptoms; the type of concomitant treatment; the frequency of treatment; the route of administration, the renal and hepatic function of the patient, and the desired effect. A physician or veterinarian can determine and prescribe the effective amount of the drug necessary to prevent, counteract, or arrest the progression of the disorder.

[0130]

[0134] In some embodiments, the pharmaceutical composition of the present disclosure can be formulated to administer a compound provided herein, or a pharmaceutically acceptable salt thereof, at a dosage between 0.001 and 1000 mg / kg body weight per day, such as 0.01 to 800 mg / kg body weight per day, 0.01 to 700 mg / kg body weight per day, 0.01 to 600 mg / kg body weight per day, 0.01 to 500 mg / kg body weight per day, 0.01 to 400 mg / kg body weight per day, 0.01 to 300 mg / kg body weight per day, 0.1 to 200 mg / kg body weight per day, 0.1 to 150 mg / kg body weight per day, 0.1 to 100 mg / kg body weight per day, 0.5 to 100 mg / kg body weight per day, 0.5 to 80 mg / kg body weight per day, 0.5 to 60 mg / kg body weight per day, 0.5 to 50 mg / kg body weight per day, 1 to 50 mg / kg body weight per day, 1 to 45 mg / kg body weight per day, 1 to 40 mg / kg body weight per day, 1 to 35 mg / kg body weight per day, 1 to 30 mg / kg body weight per day, 1 to 25 mg / kg body weight per day. In some examples, a dosage level below the lower limit of the above range may be sufficient, and in some examples, even higher dosages may be used without causing any harmful side effects, provided that such higher dosages are initially divided into several smaller dosages for administration throughout the day. For further information regarding the route of administration and dosing schedule, see Chapter 25.3 of Volume 5 of Comprehensive Medicinal Chemistry (Corwin Hansch; Chairman of Editorial Board), Pergamon Press 1990. This is hereby expressly incorporated herein by reference.

[0131]

[0135] In some embodiments, the pharmaceutical composition of the present disclosure can be formulated as a single dosage form. The amount of the compound provided herein in the single dosage form will vary depending on the subject being treated and the particular mode of administration.

[0132]

[0136] In some embodiments, a dosage form suitable for administration can contain from about 1 mg to about 1000 mg of the active ingredient per dosage unit. In these pharmaceutical compositions, the active ingredient will typically be present in an amount of from about 0.1 to 95% by weight, based on the total weight of the composition.

[0133]

[0137] In some embodiments, the pharmaceutical compositions of the present disclosure can be formulated as short-acting, immediate-release, long-acting, and sustained-release. Thus, the pharmaceutical formulations of the present disclosure can also be formulated for controlled release or sustained release.

[0134]

[0138] In some embodiments, the dosage of the compounds provided herein or the pharmaceutical compositions provided herein is administered to a subject daily, every other day, every 2 to 3 days, every 3 days, once a week, twice a week, three times a week, or once every two weeks. If desired, the active compound of the effective daily dose can be administered as separate partial doses at appropriate intervals throughout the day, 2, 3, 4, 5, or 6 or more times, optionally in unit dosage forms. In some embodiments, the dosage of the compounds provided herein or the pharmaceutical compositions provided herein is administered for 2 days, 3 days, 5 days, 7 days, 14 days, 21 days, 1 month, 2 months, 2.5 months, 3 months, 4 months, 5 months, or 6 months or more.

[0135]

[0139] In a further aspect, there is also provided a veterinary composition comprising one or more molecules or compounds of the present disclosure or a pharmaceutically acceptable salt thereof and a veterinary carrier. A veterinary carrier is a material useful for administering the composition and can be a solid, liquid, or gaseous material that is otherwise inert or acceptable in the art of veterinary medicine and compatible with the active ingredient. These veterinary compositions can be administered parenterally, orally, or by any other desired route.

[0136]

[0140] A pharmaceutical composition or veterinary composition can be packaged in various ways depending on the method used to administer the drug. For example, as a commercial item, a container containing the composition in a suitable form can be mentioned. Suitable containers are well known to those skilled in the art and include materials such as bottles (plastic and glass), sachets, ampoules, plastic bags, and metal cylinders. The container can also include an anti-tampering set to prevent inadvertent access to the contents of the package. In addition, the container is labeled to describe the contents of the container. The label can also include appropriate cautions. The composition can also be packaged in single-dose containers or multi-dose containers, such as sealed ampoules and vials, and stored in a freeze-dried (lyophilized) state that requires only the addition of a sterile liquid carrier, such as water, immediately before use for injection. Ready-to-prepare injection solutions and injection suspensions are prepared from sterile powders, granules, and tablets as described above.

[0137]

[0141] In some embodiments, the pharmaceutical compositions of the disclosure comprising one or more of the compounds provided herein or pharmaceutically acceptable salts thereof further comprise one or more additional therapeutically active agents.

[0138]

[0142] The additional therapeutically active agents have complementary activities so as not to adversely affect each other with the compounds provided herein. Such agents are combined in effective amounts for the intended purpose and are present appropriately.

[0139]

[0143] In certain embodiments, the additional therapeutic agent is selected from beta-blockers, alpha-agonists, carbonic anhydrase inhibitors, prostaglandin-like compounds, miotics or cholinergic agents, or epinephrine compounds.

[0140]

[0144] Beta blockers reduce the production of aqueous humor. Examples include levobunolol (BETAGAN®), timolol (BETIMOL®, TIMOPTIC®), betaxolol (BETOPTIC®), and metipranolol (OPTIPRANOLOL®).

[0141]

[0145] Alpha-agonists reduce the production of aqueous humor and increase its drainage. Examples include apraclonidine (IOPIDINE®) and brimonidine (ALPHAGAN®).

[0142]

[0146] Carbonic anhydrase inhibitors reduce the production of aqueous humor. Examples include dorzolamide (TRUSOPT®) and brinzolamide (AZOPT®).

[0143]

[0147] Prostaglandins and prostaglandin-like compounds increase the outflow of aqueous humor. Examples include latanoprost (XALATAN®), bimatoprost (LUMIGAN®), and travoprost (TRAVATAN™).

[0144]

[0148] Miotic agents or cholinergic agonists increase the outflow of aqueous humor. Examples include pilocarpine (ISOPTO CARPINE®, PILOPINE®) and carbachol (ISOPTO CARBACHOL®).

[0145]

[0149] Epinephrine compounds, such as dipivefrin (PROPINE®), also increase the outflow of aqueous humor.

[0150] Additional therapeutic or agent can be administered simultaneously or sequentially with the compounds provided herein. Sequential administration includes administration before or after the compounds provided herein. In some embodiments, the additional therapeutic or agent may be administered in the same composition as the compounds provided herein. In other embodiments, there may be a time interval between the administration of the additional therapeutic agent and the administration of the compounds provided herein.

[0146]

[0151] In some embodiments, administering an additional therapeutic agent together with the compounds provided herein may reduce the dosage of other therapeutic agents and / or enable administration at less frequent intervals.

[0147] Method of treatment

[0152] The compounds of the present disclosure and pharmaceutical compositions containing the same can inhibit ROCK, and thus may be useful for inhibiting ROCK activity in a subject in need of inhibition of ROCK activity and for preventing or treating ROCK-related disorders.

[0148]

[0153] In a further aspect, the present disclosure provides a method of treating a ROCK-related disorder, the method comprising administering to a subject in need thereof an effective amount of a compound provided herein or a pharmaceutically acceptable salt or pharmaceutical composition thereof.

[0149]

[0154] As used herein, the terms "treating," "treatment," or "therapy" shall have their ordinary meaning of addressing a disease to wholly or partially alleviate one, several, or all of the symptoms of the disease or to ameliorate or compensate for the underlying pathology, thereby achieving a beneficial or desired clinical outcome. For the purposes of the present disclosure, a beneficial or desired clinical outcome includes, but is not limited to, alleviation of symptoms, reduction in the degree of the disease, stabilization of the disease state (i.e., not exacerbating), delay or slowing of disease progression, improvement or alleviation of the disease state, and remission (partial or complete), whether detectable or undetectable. "Treating" can also mean extending the survival period compared to the expected survival period if not receiving treatment. Those in need of treatment include those already having a condition or disorder, those having a tendency to have a condition or disorder, and those in whom a condition or disorder should be prevented.

[0150]

[0155] As used herein, the terms "preventing," "prevention," or "prophylaxis" shall have their ordinary meaning and include primary prevention to prevent the onset of a disease and secondary prevention where the disease has already occurred and the patient is temporarily or continuously protected against disease exacerbation or worsening, or the onset of new symptoms associated with the disease.

[0151]

[0156] In some embodiments, the compounds or their pharmaceutically acceptable salts and compositions provided herein can be used for the treatment of various ROCK-related disorders including cancer, cardiovascular diseases, autoimmune diseases, eye diseases, metabolic syndrome, respiratory distress syndrome, kidney diseases, overactive bladder, epilepsy, migraine, diabetes, high altitude pulmonary edema, mental disorders, and the like.

[0152]

[0157] In certain embodiments, the compounds or pharmaceutically acceptable salts and compositions thereof provided herein can be used to treat glaucoma, ocular hypertension, eye diseases including retinal diseases, bone disorders including osteoporosis and osteoarthritis, cerebral vasospasm, coronary artery spasm, hypertension, pulmonary hypertension, high altitude essential hypertension, sudden death syndrome, angina, myocardial infarction, restenosis, stroke, hypertensive vascular diseases, heart failure, cardiac allograft vasculopathy, atherosclerosis, arterial occlusion, peripheral arterial disease, peripheral circulatory disorders, vascular diseases including vein graft lesions, lung diseases including chronic obstructive pulmonary disease (COPD) and asthma, spinal cord injury, dementia, Alzheimer's disease, Parkinson's disease, neurodegeneration, multiple sclerosis, depression, attention deficit hyperactivity disorder and neuropathic pain, multiple sclerosis, neuropathies including amyotrophic lateral sclerosis, neovascular disorders, astrocytes, breast, cervical, colorectal, endometrial, esophageal, gastric, head and neck, hepatocellular, laryngeal, lung, oral, ovarian, prostate and thyroid cancers and sarcomas, obesity, urological 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, and myocardial protection.

[0153]

[0158] In certain embodiments, the eye disease that can be treated with the compounds or pharmaceutically acceptable salts and compositions thereof provided herein is glaucoma.

[0159] The concentration and route of administration to a subject will vary depending on the ROCK-related disorder to be treated. In certain embodiments, administration is via a route selected from the group consisting of parenteral administration, intraperitoneal administration, intradermal administration, intracardiac administration, intracerebroventricular administration, intracranial administration, intrathecal administration, intrasynovial administration, subarachnoid administration, intramuscular injection, intravitreal injection, intravenous injection, intraarterial injection, oral administration, buccal administration, sublingual administration, transdermal administration, topical administration, intratracheal administration, rectal administration, subcutaneous administration, and transocular administration.

Example

[0154]

[0160] The general methods of the present disclosure are further described below. The compounds of the present disclosure can be prepared by methods known in the art. The following describes a detailed method for preparing preferred compounds of the present disclosure. However, the description is in no way intended to limit the method for preparing the compounds of the present disclosure.

[0155] Examples of Synthesis

[0161] For illustrative purposes, the following examples are included. The examples provided herein illustrate the synthesis of the compounds disclosed herein and the intermediates used to prepare those compounds. However, of course, these examples are not intended to limit the present disclosure and are only intended to suggest ways of practicing the present disclosure. Those skilled in the art will recognize that the described chemical reactions can be readily adapted to prepare a plurality of other compounds of the present disclosure, and that other methods of preparing the compounds of the present disclosure are considered to be within the scope of the present disclosure. For example, the synthesis of compounds not exemplified by the present disclosure can be accomplished by obvious modifications by those skilled in the art, such as appropriately protecting interfering groups, utilizing other suitable reagents, constructing blocks known in the art other than those described, and / or making routine changes to reaction conditions. In addition, those skilled in the art will understand that each step described herein or in separate batches of the compound may be combined. Alternatively, other reactions disclosed herein or known in the art will be recognized as having applicability for preparing other compounds of the present disclosure. Accordingly, the following description is not intended to limit the scope of the present disclosure, but rather is defined by the claims appended hereto.

[0156]

[0162] The abbreviations of the chemical substances used in the synthesis of the compounds provided herein are listed below.

[0157]

Table 2-1

[0158]

Table 2-2

[0159]

Table 2-3

[0160] Synthesis of Intermediate Synthesis of Intermediate I-1

[0161]

Chemical formula

[0162]

[0163] Step 1: A solution of p-cyanomethylbenzoic acid I-1a (20 g, 124.10 mmol, 1 equiv) and CDI (22.14 g, 136.51 mmol, 1.1 equiv) in THF (50 mL) was stirred at room temperature for 3 h under a nitrogen atmosphere. To the mixture, NaBH 2 in H 4 (14.08 g, 372.30 mmol, 3 equiv) in O (45 mL) was added dropwise 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 at room temperature. The resulting mixture was extracted with ethyl acetate (3 × 500 mL). The combined organic layers were dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica column chromatography eluting with CH 2 Cl 2 / EtOAc (8:1, v / v) to give 2-[4-(hydroxymethyl)phenyl]acetonitrile I-1b (10.9 g, 59.67%, crude) as a pale yellow liquid. 1 H NMR (CD 3 OD, 400 MHz) δ 3.87 - 3.91 (2H, m), 4.62 (2H, s), 7.37 (4H, q).

[0164] Step 2: To a stirred mixture of alcohol I-1b (10.9 g, 74.06 mmol, 1 equiv), triethylamine (22.48 g, 222.18 mmol, 3 equiv), and DMAP (0.09 g, 0.741 mmol, 0.01 equiv) in DCM (100 mL) was added TBSCl (13.39 g, 88.87 mmol, 1.2 equiv) portionwise at 0 °C under a nitrogen atmosphere. The resulting mixture was stirred at room temperature for 4 h under a nitrogen atmosphere. The resulting mixture was diluted with water (260 mL). The resulting mixture was extracted with CH 2 Cl 2 (3 × 100 mL), and the combined organic layers were dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC (PE / EA = 6:1, v / v) to give 2-(4-(((tert-butyldimethylsilyl)oxy)methyl)phenyl)acetonitrile I-1c (14.0 g, 72.31%) as a pale yellow liquid. 1 H NMR (CD 3 OD, 400 MHz) δ 0.13 (6H, s), 0.96 (9H, s), 3.89 (2H, s), 4.77 (2H, s), 7.31 - 7.40 (4H, m).

[0165] Step 3: To a stirred solution of nitrile I-1c (14.0 g, 53.55 mmol, 1 equiv) in THF (20 mL) was added sodium hydride (1.93 g, 80.33 mmol, 1.5 equiv) portionwise. The resulting mixture was stirred at 0 °C for 1 h under a nitrogen atmosphere. Dimethyl carbonate (19.29 g, 214.20 mmol, 4 equiv) was added portionwise 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 resulting mixture was extracted with ethyl acetate (3 × 250 mL), and the combined organic layers were dried over anhydrous Na 2 SO 4It was dehydrated. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica column chromatography (PE / EA = 6:1, volume / volume) to obtain methyl 2-(4-{[(tert-butyldimethylsilyl)oxy]methyl}phenyl)-2-cyanoacetate I-1d (12.6 g, 73.65%) as a pale yellow liquid. 1 H NMR (CD 3 OD, 400 MHz) δ 0.13 (6H, s), 0.97 (9H, s), 3.78 (3H, s), 4.78 (2H, s), 7.37 - 7.47 (4H, m).

[0166] Step 4: In a 250 mL round-bottom flask, at room temperature, ester I-1d (12.6 g, 39.44 mmol, 1 equivalent) and Boc 2 O (34.44 g, 157.76 mmol, 4 equivalents), CoCl 2 ·6H 2 O (24.42 g, 102.55 mmol, 2.6 equivalents) were added to methanol (150 mL). NaBH 4 (11.94 g, 315.53 mmol, 8 equivalents) was added to the above mixture at 0 °C. The resulting mixture was stirred at room temperature for 15 hours under a nitrogen atmosphere. The combined organic layers were dehydrated with anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica column chromatography eluted with PE / EA (0 - 5.5%) to obtain methyl 3-[(tert-butoxycarbonyl)amino]-2-(4-{[(tert-butyldimethylsilyl)oxy]methyl}phenyl)propanoate I-1e (7.1 g, 42.51%) as a pale yellow oil. 1 H NMR (DMSO-d 6 , 400 MHz) δ 0.08 (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).

[0167] Step 5: Ester I-1e (10.1 g, 23.84 mmol, 1 equiv) in methanol (10 mL) and H 2 A solution of LiOH·H 2 O (2.00 g, 47.68 mmol, 2 equiv) in H + O (6 mL) was stirred overnight at 60 °C under a nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure. The mixture was acidified to pH 6 with HCl (aqueous solution). The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, methanol in water, concentration gradient from 10% to 50% in 40 min; detector, UV 220 nm. Thereby, 3-[(tert-butoxycarbonyl)amino]-2-[4-(hydroxymethyl)phenyl]propanoic acid I-1 (3.58 g, 50.84%) was obtained as an off-white solid. LCMS: m / z (ESI), [M+Na]

[0163] Synthesis of Intermediate I-2

[0164]

Chemical formula

[0165]

[0168] Step 1: To a solution of 2-(3-methoxyphenyl)acetonitrile I-2a (3 g, 20.38 mmol, 1 equiv) in THF (50 mL) was added sodium hydride (0.54 g, 22.42 mmol, 1.1 equiv) at 0 °C. The mixture was stirred for 50 min. Dimethyl carbonate (14.69 g, 163.07 mmol, 8 equiv) was added, and the mixture and the resulting mixture were warmed to room temperature and stirred for 2 h. The reaction mixture was quenched with water (50 mL) and extracted with DCM (3 × 50 mL). The combined organic layers were dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica column chromatography eluting with PE / EA (12:1, volume / volume) to give methyl 2-cyano-2-(3-methoxyphenyl)acetate I-2b (2.84 g, 67.89%) as a yellow oil. 1 H NMR (400 MHz, DMSO-d 6) δ 3.76 (d, 6H), 5.63 (s, 1H), 6.81 - 7.11 (m, 3H), 7.39 (t, 1H).

[0169] Step 2: In a mixture of ester I-2b (800 mg, 3.90 mmol, 1 equivalent), Boc 2 O (1701.62 mg, 7.80 mmol, 2 equivalents), CoCl 2 ·6H 2 O (1113.06 mg, 4.68 mmol, 1.2 equivalents) and NaBH 4 (368.72 mg, 9.75 mmol, 2.5 equivalents) in methanol (20 mL) was stirred at room temperature overnight under a nitrogen atmosphere. The resulting mixture was diluted with water (30 mL) and extracted with CH 2 Cl 2 (3 × 40 mL), and dried over anhydrous Na 2 SO 4 . 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)propanoate I-2c (504 mg, 41.79%) as a pale yellow oil. LCMS: m / z (ESI), [M+Na] + = 332.00.

[0166]

[0170] Step 3: To a 50 mL round-bottom flask were added ester I-2c (760 mg, 2.46 mmol, 1 equivalent) and LiOH (117.67 mg, 4.91 mmol, 2.0 equivalents) in THF (3 mL) and water (3 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 hours. The resulting mixture was concentrated under reduced pressure. The mixture was acidified to pH 5 with HCl (1 M). The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, methanol in water, concentration gradient from 0% to 100% in 50 minutes; detector, UV254 nm. The resulting mixture was concentrated under vacuum to give 3-[(tert-butoxycarbonyl)amino]-2-(3-methoxyphenyl)propanoic acid I-2 (200 mg, 27.57%) as a yellowish brown solid. LCMS: m / z (ESI), [M- t Bu]+ = 240.25.

[0167] Synthesis of Intermediate I-3

[0168]

Chem.

[0169]

[0171] Step 1: To a solution of 2-(3-bromophenyl)acetonitrile I-3a (10 g, 51.01 mmol, 1 equiv) in THF (200 mL) at 0 °C was added sodium hydride (4.08 g, 102.02 mmol, 2 equiv, 60%). The mixture was stirred for 40 minutes. Dimethyl carbonate (18.38 g, 204.03 mmol, 4 equiv) was added and the mixture was warmed to room temperature 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 × 200 mL). The combined organic layers were washed with brine (2 × 200 mL) and dried over anhydrous Na 2 SO 4 4. 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), [M-H] - = 253.70. 1 H NMR (DMSO-d 6 6, 400 MHz) δ 3.43 (3H, s), 6.67 (1H, ddd), 6.94 (1H, t), 7.39 (1H, d), 7.86 (1H, s).

[0172] Step 2: To a stirred mixture of ester I-3b (5.5 g, 21.65 mmol, 1 equiv), CoCl 2 2·6H 2 2O (15.45 g, 64.94 mmol, 3 equiv) and Boc 2 2O (14.17 g, 64.94 mmol, 3 equiv) in methanol (100 mL) at 0 °C under an air atmosphere was added NaBH 4(4.91 g, 129.88 mmol, 6 eq) was added dropwise, the mixture was warmed to room temperature and stirred for 2 h. The resulting mixture was quenched with water (200 mL) at 0 °C. The resulting mixture was filtered. The filtered cake was washed with DCM (4 × 100 mL). The filtrate was extracted with CH 2 Cl 2 (2 × 300 mL). The combined organic layers were dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica column chromatography eluting with PE / EA (10:1, v / v) to give methyl 2-(3-bromophenyl)-3-[(tert-butoxycarbonyl)amino]propanoate I-3c (4 g, 30.95%) as a yellow oil. LCMS: m / z (ESI), [M + H - t Bu] + = 303.85.

[0170]

[0173] Step 3: A mixture of Pd(PPh 3 ) 4 (0.16 g, 0.14 mmol, 0.05 eq), Zn(CN) 2 (0.20 g, 1.68 mmol, 0.6 eq) and ester I-3c (1 g, 2.79 mmol, 1 eq) in DMF (15 mL) was stirred at 80 °C for 4 h under a nitrogen atmosphere. The resulting mixture was diluted with water (50 mL). The resulting mixture was extracted with ethyl acetate (3 × 50 mL). The combined organic layers were washed with brine (3 × 70 mL) and dried over anhydrous Na 2 SO 4 . 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)propanoate I-3d (400 mg, 47.08%) as a white solid. LCMS: m / z (ESI), [M + H - t Bu] + = 248.95. 1 H NMR (400 MHz, DMSO-d 6) δ 1.31 (9H, s), 3.33 - 3.42 (1H, m), 3.46 - 3.58 (1H, m), 3.63 (3H, s), 3.96 (1H, t), 6.96 (1H, t), 7.47 - 7.65 (2H, m), 7.73 (1H, d), 7.77 (1H, d).

[0174] Step 4: LiOH·H 2 O (86.17 mg, 2.05 mmol, 2.5 equiv) and nitrile I - 3d (250 mg, 0.82 mmol, 1 equiv) in THF (5 mL) and H 2 O (1 mL) were stirred at room temperature for 2 h under an air atmosphere. The resulting mixture was concentrated under reduced pressure to give 3 - [(tert - butoxycarbonyl)amino] - 2 - (3 - cyanophenyl)propanoic acid I - 3 (200 mg, 83.87%) as a white solid. LCMS: m / z (ESI), [M + H] + = 291.00.

[0171] Synthesis of Intermediate I - 4

[0172]

Chemical Structure

[0173]

[0175] Step 1: To a stirred mixture of 2 - (4 - bromophenyl) - 3 - [(tert - butoxycarbonyl)amino]propanoic acid (2.0 g, 5.81 mmol, 1 equiv) and DMAP (141.97 mg, 1.16 mmol, 0.2 equiv) in DCM (10 mL) and methanol (1.0 mL, 24.70 mmol, 4.25 equiv) was added EDCI (2227.74 mg, 11.620 mmol, 2 equiv) portionwise at room temperature under a nitrogen atmosphere. The resulting mixture was stirred overnight at room temperature under a nitrogen atmosphere. After the reaction, the resulting mixture was diluted with water (40 mL). The resulting mixture was extracted with ethyl acetate (3 × 30 mL). The combined organic layers were washed with water (3 × 20 mL) and anhydrous Na 2 SO 4It was dehydrated. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica column chromatography eluted with PE / EA (4:1, volume / volume) to obtain methyl 2-(4-bromophenyl)-3-[(tert-butoxycarbonyl)amino]propanoate I-4a (1.58 g, 75.91%) as a white solid. LCMS: m / z (ESI), [M-t-Bu] + = 301.80.

[0174]

[0176] Step 2: To a stirred mixture of ester I-4a (1.48 g, 4.13 mmol, 1 equiv) and Zn(CN) 2 (494.81 mg, 4.21 mmol, 1.02 equiv) in DMF (4 mL), Pd(PPh 3 ) 4 (477.42 mg, 0.41 mmol, 0.1 equiv) was added portionwise under a nitrogen atmosphere at room temperature. The resulting mixture was stirred at 80 °C for 3 h under a nitrogen atmosphere. After cooling to room temperature, the resulting mixture was diluted with water (25 mL). The resulting mixture was extracted with ethyl acetate (3 × 30 mL). The combined organic layers were dehydrated with anhydrous Na 2 SO 4 . 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 using PE / EA (3:1, volume / volume) to obtain methyl 3-[(tert-butoxycarbonyl)amino]-2-(4-cyanophenyl)propanoate I-4b (1.1 g, 87.48%) as a yellow oil. LCMS: m / z (ESI), [M-t-Bu] + = 249.30.

[0175]

[0177] Step 3: To a stirred mixture of nitrile I-4b (300 mg, 0.99 mmol, 1 equiv) and (Boc) 2 O (430.26 mg, 1.97 mmol, 2 equiv), CoCl 2 ·6H 2 O (469.05 mg, 1.97 mmol, 2 equiv) in methanol (10 mL), NaBH 4(298.32 mg, 7.89 mmol, 8 equivalents) was added little by little. The resulting mixture was stirred at 25 °C for 3 hours under a nitrogen atmosphere. The reaction was quenched with ice water (30 mL) at 0 °C. The resulting mixture was filtered, and the filtered cake was washed with methanol (3 × 20 mL). The filtrate was concentrated under reduced pressure. The resulting mixture was extracted with ethyl acetate (3 × 30 mL). The combined organic layers were washed with water (3 × 10 mL) and dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC using PE / EA (5:1, volume / volume) to obtain methyl 3-[(tert-butoxycarbonyl)amino]-2-(4-{[(tert-butoxycarbonyl)amino]methyl}phenyl)propanoate I-4c (170 mg, 42.22%) as a yellow oil. LCMS: m / z (ESI), [M+Na] + = 431.10.

[0176]

[0178] Step 4: To a stirred mixture of ester I-4c (160 mg, 0.39 mmol, 1 equivalent) in methanol (4.0 mL), LiOH·H 2 O (65.74 mg, 1.57 mmol, 4.0 equivalents) in water (2.0 mL) was added dropwise at 25 °C under a nitrogen atmosphere. The resulting mixture was stirred at 60 °C for 3 hours under a nitrogen atmosphere. 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 reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, MeCN in water, 10% to 50% concentration gradient in 10 minutes; detector, UV 254 nm, held at 30% for 20 minutes. After evaporation of the solvent, 3-[(tert-butoxycarbonyl)amino]-2-(4-{[(tert-butoxycarbonyl)amino]methyl}phenyl)propanoic acid I-4 (90 mg, 58.25%) was obtained as a white solid. LCMS: m / z (ESI), [M-H] - = 393.15.

[0177] Synthesis of Intermediate I-5

[0178]

Chemical Structure

[0179]

[0179] Step 1: In a 20 mL vial, at room temperature, 3-iodonitrobenzene I-5a (12.56 g, 50.46 mmol, 1 equiv) in DMSO (100 mL), K 2 CO 3 (20.92 g, 151.38 mmol, 3 equiv), CuI (9.61 g, 50.46 mmol, 1 equiv) and methyl cyanoacetate (5 g, 50.46 mmol, 1 equiv) were added. The resulting mixture was stirred at 120 °C for 3 h under a nitrogen atmosphere. The resulting mixture was diluted with EA (200 mL) and washed with saturated NaHCO 3 (aqueous solution) 1 × 200 mL, water (2 × 200 mL) and saturated brine (1 × 200 mL). The organic layer was concentrated under reduced pressure. The residue was purified by silica column chromatography eluting with PE / EA (1:1, v / v). The residue was purified by silica column chromatography eluting with PE / EA (25:1) to give methyl 2-cyano-2-(3-nitrophenyl)acetate I-5b (3.0 g, 27.00%) as a red oil. 1 H NMR (DMSO-d 6 , 400 MHz) δ 3.77 (3H, s), 6.01 (1H, s), 7.79 (1H, t), 7.93 (1H, d), 8.30 (1H, dd), 8.34 (1H, t).

[0180] Step 2: NaBH 4 (604.77 mg, 15.98 mmol, 8 equiv) was added to the ester I-5b (440 mg, 2.00 mmol, 1 equiv) at 0 °C, CoCl 2 ·6H 2 O (1.24 g, 5.20 mmol, 2.6 equiv) and Boc 2O (1.74 g, 7.99 mmol, 4 equiv) was added to a mixture in methanol (10 mL). The resulting mixture was stirred at room temperature for 1 h. The resulting mixture was quenched with water (50 mL), filtered, and the filtered cake was washed with DCM (3 × 10 mL). The filtrate was concentrated under reduced pressure. The resulting mixture was extracted with DCM (3 × 50 mL). The organic layer was concentrated under reduced pressure. The residue was purified by preparative TLC using PE / EA (2:1, v / v) to give methyl 3-[(tert-butoxycarbonyl)amino]-2-{3-[(tert-butoxycarbonyl)amino]phenyl}propanoate I-5c (300 mg, 38.06%) as a yellow oil. LCMS: m / z (ESI), [M+H] + = 395.20. 1 H NMR (CD 3 OD, 400 MHz) 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).

[0181] Step 3: In a 50 mL round-bottom flask, ester I-5c (300 mg, 0.76 mmol, 1 equiv) and LiOH (36.43 mg, 1.52 mmol, 2 equiv) in THF (2.5 mL) and water (2.5 mL) were added at room temperature. The resulting mixture was stirred at room temperature for 1.5 h. The mixture was acidified to pH 5 with HCl (1 M). The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, methanol in water, 0% to 20% concentration gradient in 30 min; detector, UV254 nm. The resulting mixture was concentrated under vacuum to give 3-[(tert-butoxycarbonyl)amino]-2-{3-[(tert-butoxycarbonyl)amino]phenyl}propanoic acid I-5 (200 mg, 69.12%) as a yellow oil. 1 H NMR (CD 3OD, 400 MHz) δ 1.42 (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). Synthesis of Intermediate I-6

[0180]

Chem.

[0181]

[0182] Step 1: In a 250 mL round-bottom flask, ethyl cyanoacetate (2.26 g, 19.98 mmol, 1 equiv), oxan-3-one I-6a (2 g, 19.98 mmol, 1 equiv), etidine (5.06 g, 19.98 mmol, 1 equiv), L-proline (0.46 g, 4.00 mmol, 0.2 equiv), and methanol (100 mL) were added at room temperature. The resulting mixture was stirred at 25 °C for 15 h under a nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica column chromatography eluting with PE / EA (2:1, v / v) to give ethyl 2-cyano-2-(oxan-3-yl)acetate I-6b (3.2 g, 81.22%) as a yellow solid. 1 H NMR (CDCl 3 , 400 MHz) δ 1.35 (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).

[0183] Step 2: In a 250 mL round-bottom flask, ester I-6b (3.1 g, 15.72 mmol, 1 equiv) and Boc 2 O (6.86 g, 31.43 mmol, 2 equiv) and CoCl 2 ·6H 2 O (7.48 g, 31.43 mmol, 2 equiv) were added in methanol (80 mL) at 0 °C. NaBH4 (2.97 g, 78.59 mmol, 5 equiv) was added portionwise to the above mixture at 0 °C. The resulting mixture was stirred at room temperature for 12 h under a nitrogen atmosphere. The reaction was quenched at 0 °C by the addition of water (200 mL). The resulting mixture was filtered and the filter cake was washed with DCM (6 × 50 mL). The aqueous layer was extracted with CH 2 Cl 2 (3 × 200 mL). The resulting mixture was concentrated under reduced pressure. The residue was purified by silica column chromatography eluting with PE / EA (2:1, v / v) to afford ethyl 3-[(tert-butoxycarbonyl)amino]-2-(oxan-3-yl)propanoate I-6c (3 g, 63.33%) as a yellow oil. LCMS: m / z (ESI), [M + H - t Bu] + = 246.05. 1 H NMR (CDCl 3 , 400 MHz) δ 1.26 - 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).

[0184] Step 3: In a 50 mL round-bottom flask, at room temperature, ester I-6c (1 g, 3.32 mmol, 1 equiv) and LiOH·H 2 O (158.93 mg, 6.64 mmol, 2 equiv) and H 2O (4 mL) and methanol (8 mL) were added. The resulting mixture was stirred at 60 °C for 15 h under a nitrogen atmosphere. The mixture was acidified to pH 5 with HCl (1 M). The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, methanol in water, concentration gradient from 0% to 30% in 40 min; detector, UV 220 nm. The resulting mixture was concentrated under reduced pressure. The obtained solid was dried by lyophilization. Thereby, 3-[(tert-butoxycarbonyl)amino]-2-(oxan-3-yl)propanoic acid I-6 (650 mg, 71.67%) was obtained as a white solid. LCMS: m / z (ESI), [M+H- t Bu] + = 218.15. 1 H NMR (CDCl 3 , 400 MHz) δ 1.48 (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). Synthesis of Intermediate I-7

[0182]

Chemical Structure

[0183]

[0185] Step 1: In a 250 mL round-bottom flask, 2-(pyridin-3-yl)acetonitrile I-7a (6 g, 50.79 mmol, 1 equiv) and THF (40 mL, 493.71 mmol), sodium hydride (2.23 g, 55.87 mmol, 1.1 equiv, 60%) were added at 0 °C. The resulting mixture was stirred at room temperature for 1 h, and dimethyl carbonate (36.60 g, 406.30 mmol, 8 equiv) was added to the above mixture. The resulting mixture was stirred at room temperature for 4 h. The resulting mixture was extracted with CH 2 Cl 2 / methanol = 20:1 (3 × 30 mL). The combined organic layers were dried over anhydrous Na 2 SO 4It was dehydrated. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica column chromatography eluting with PE / EA (1:1, volume / volume) to obtain methyl 2-cyano-2-(pyridin-3-yl)acetate I-7b (7.4 g, 54.83%) as a yellowish brown solid. LCMS: m / z (ESI), [M+H] + = 177.25.

[0184]

[0186] Step 2: In a 100 mL round-bottom flask, at 0 °C, ester I-7b (2 g, 11.35 mmol, 1 equivalent) in methanol (40 mL), CoCl 2 ·6H 2 O (8.10 g, 34.06 mmol, 3 equivalents) and Boc 2 O (4.96 g, 22.70 mmol, 2 equivalents) were added, and NaBH 4 (3.44 g, 90.82 mmol, 8 equivalents) was added. The resulting mixture was stirred at 0 °C for 2 hours. The reaction was quenched with water at room temperature. The resulting mixture was extracted with CH 2 Cl 2 (3 × 30 mL) and dehydrated with anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC using DCM / EA (1:1, volume / volume) to obtain methyl 3-[(tert-butoxycarbonyl)amino]-2-(pyridin-3-yl)propanoate I-7c (310 mg, 8.14%) as a yellow solid. LCMS: m / z (ESI), [M+H] + = 281.15.

[0185]

[0187] Step 3: In a 50 mL round-bottom flask, ester I-7c (300 mg, 1.07 mmol, 1 equiv) was added at room temperature, followed by LiOH (51.26 mg, 2.14 mmol, 2 equiv), THF (5 mL) and water (5 mL). The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, MeCN in water, concentration gradient from 10% to 50% in 40 min; detector, UV 220 nm. This gave 3-[(tert-butoxycarbonyl)amino]-2-(pyridin-3-yl)propanoic acid I-7 (140 mg, 41.16%) as a yellow solid. LCMS: m / z (ESI), [M+H] + = 267.05.

[0186] Synthesis of Intermediate I-8

[0187]

Chem.

[0188]

[0188] Step 1. In a 250 mL round-bottom flask, atropic acid I-8a (8 g, 54.00 mmol, 1 equiv) and ethanol (220 mL), H 2 SO 4 (0.5 mL, 9.38 mmol, 0.17 equiv) were added at room temperature. The resulting mixture was stirred at 80 °C overnight. After cooling to room temperature, the mixture was neutralized to pH 8 with saturated NaHCO 3 (aqueous solution). The resulting mixture was diluted with water (70 mL). The aqueous layer was extracted with ethyl acetate (3 × 100 mL). The resulting mixture was concentrated under reduced pressure. The residue was purified by silica column chromatography eluting with PE / EA (10:1, v / v) to give ethyl 2-phenylprop-2-enoate I-8b (7.5 g, 78.82%) as a colorless liquid. 1 H NMR (400 MHz, DMSO-d 6 ) δ 1.3 (t, 3H), 4.2 (q, 2H), 6.0 (d, 1H), 6.2 (d, 1H), 7.2 - 7.5 (m, 5H).

[0189] Step 2: A solution of ester I-8b (250 mg, 1.50 mmol, 1 equiv) and 1-methylpiperazine (568.42 mg, 5.68 mmol, 4 equiv) in DMF (3 mL) was stirred at room temperature for 1 h under a nitrogen atmosphere. The residue was dissolved in PE (20 mL). The combined organic layers were washed with water (3 × 10 mL) and dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure to afford ethyl 3-(4-methylpiperazin-1-yl)-2-phenylpropanoate I-8c (330 mg, 84.16%) as an off-white solid. LCMS: m / z (ESI), [M+H] + = 277.25.

[0189]

[0190] Step 3: To a stirred solution of piperazine I-8c (500 mg, 1.81 mmol, 1 equiv) in methanol (10 mL) and H 2 O (5 mL), LiOH (173.31 mg, 7.24 mmol, 4 equiv) was added dropwise at room temperature under an air atmosphere. The resulting mixture was stirred at room temperature for 3 h under an air atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, MeCN in water, gradient from 20% to 30% in 10 min; detector, UV 254 nm to afford 3-(4-methylpiperazin-1-yl)-2-phenylpropanoic acid I-8 (220 mg, 48.97%) as a white solid. LCMS: m / z (ESI), [M+H] + = 249.15.

[0190] Synthesis of Intermediate I-9

[0191]

Chem.

[0192]

[0191] Step 1: To a stirred mixture of ethyl phenylacetate I-9a (1000 mg, 6.09 mmol, 1 equiv) and tert-butyl 4-iodopiperidine-1-carboxylate I-9b (2.84 g, 9.14 mmol, 1.5 equiv) in DMF (10 mL) was added t-BuOK (1.37 g, 12.18 mmol, 2 equiv) portionwise at 0 °C under an air atmosphere. The resulting mixture was stirred at 25 °C for 16 h under an air atmosphere. The reaction was quenched with saturated NH 4 Cl (aqueous solution) at 0 °C. The resulting mixture was extracted with ethyl acetate (3 × 50 mL), and dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reverse flash chromatography using the following conditions: column, silica; mobile phase, MeCN in water, 50% to 90% concentration gradient over 10 min; detector, UV 254 nm to give tert-butyl 4-(2-ethoxy-2-oxo-1-phenylethyl)piperidine-1-carboxylate (550 mg, 25.99%) as a yellow solid. 1 H NMR (400 MHz, DMSO-d 6 ) δ1.13 (4H, t), 1.37 (9H, s), 1.67 (1H, d), 2.01-2.16 (1H, m), 3.37 (4H, d), 3.83 (1H, d), 3.90-4.17 (3H, m), 7.25-7.37 (5H, m).

[0192] Step 2: To a stirred solution of tert-butyl 4-(2-ethoxy-2-oxo-1-phenylethyl)piperidine-1-carboxylate (500 mg, 1.44 mmol, 1 equiv) in DCM (6 mL) was added TFA (2 mL). The resulting mixture was stirred at room temperature for 2 h under an air atmosphere. The resulting mixture was concentrated 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.

[0193]

[0193] Step 3: To a stirred mixture of ester I-9c (350 mg, 1.42 mmol, 1 equiv) and formaldehyde (424.89 mg, 14.15 mmol, 10 equiv) in DCM (10 mL) was added DIEA (914.47 mg, 7.08 mmol, 5 equiv). The resulting mixture was stirred at room temperature for 0.5 h under an air atmosphere. To the above mixture was added NaBH(OAc) 3 (899.73 mg, 4.25 mmol, 3 equiv). The resulting mixture was stirred for an additional 2 h at room temperature. The reaction was quenched at 0 °C by the addition of water (4 mL). The resulting mixture was extracted with ethyl acetate (3 × 30 mL) and dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure. Thereby, ethyl 2-(1-methylpiperidin-4-yl)-2-phenylacetate (350 mg, 94.63%) was obtained as a yellow oil. LCMS: m / z (ESI), [M+H] + = 262.10.

[0194]

[0194] Step 4: To a stirred solution of ethyl 2-(1-methylpiperidin-4-yl)-2-phenylacetate (350 mg, 1.34 mmol, 1 equiv) in THF (4 mL) and H 2 O (1 mL) was added LiOH·H 2 O (67.43 mg, 1.61 mmol, 1.2 equiv). The resulting mixture was stirred at 70 °C for 16 h under a nitrogen atmosphere. The mixture was acidified to pH 6 with HCl (1 M). The resulting mixture was concentrated under vacuum. The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, methanol in water, 10% to 50% concentration gradient over 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. 1 H NMR (400 MHz, DMSO-d 6) δ 7.41 - 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). Synthesis of Intermediate I - 10

[0195] [Chemical formula]

[0196]

[0195] Step 1: A mixture of methyl 2 - bromo - 2 - phenylacetate I - 10a (10 g, 43.65 mmol, 1 equivalent), 1 - methyl - piperazine (13.12 g, 130.96 mmol, 3 equivalents) and triethylamine (4.42 g, 43.65 mmol, 1 equivalent) in THF (100 mL) was stirred at 70 °C for 4 hours under a nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica column chromatography eluting with CH 2 Cl 2 / methanol (15:1, v / v) to give methyl 2 - (4 - methylpiperazin - 1 - yl) - 2 - phenylacetate I - 10b (6.3 g, 58.12%) as a brownish oil. LCMS: m / z (ESI), [M + H] + = 249.15.

[0197]

[0196] Step 2: In a 50 mL round - bottom flask, at room temperature, ester I - 10b (2.7 g, 10.87 mmol, 1 equivalent) and LiOH·H 2O (0.91 g, 21.75 mmol, 2.0 equiv) was added. The resulting mixture was stirred at room temperature for 5 h under a nitrogen atmosphere. The mixture was acidified to pH 6 with HCl (1 M). The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, methanol in water, 0% to 5% concentration gradient in 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.

[0198] Synthesis of Intermediate I-11

[0199]

Chemical Structure

[0200]

[0197] Step 1: To a stirred solution of methyl 3-(cyanomethyl)benzoate I-11a (15 g, 85.62 mmol, 1 equiv) in methanol (250 mL) was added NaBH 4 (16.20 g, 428.12 mmol, 5 equiv) portionwise at 0 °C under an air atmosphere. The resulting mixture was stirred overnight at 70 °C under an air atmosphere. The mixture was cooled to room temperature. The resulting mixture was concentrated under reduced pressure. The reaction was quenched at room temperature by the addition of water (100 mL). The resulting mixture was extracted with ethyl acetate (3 × 50 mL). The combined organic layers were dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica column chromatography eluting with PE / EA (1:10, v / v) to give 2-[3-(hydroxymethyl)phenyl]acetonitrile (11.14 g, 88.40%) as a yellow oil. 1 H NMR (DMSO-d 6 , 400 MHz) δ 4.10 (2H, s), 4.52 (2H, d), 5.28 (1H, t), 7.21 (1H, d), 7.27 (1H, d), 7.32 - 7.39 (2H, m).

[0198] Step 2: To a stirred solution of 2-[3-(hydroxymethyl)phenyl]acetonitrile (11.14 g, 75.69 mmol, 1 equiv) and TBSCl (17.11 g, 113.54 mmol, 1.5 equiv) in DCM (100 mL) was added dropwise DMAP (0.92 g, 7.569 mmol, 0.1 equiv) and triethylamine (22.98 g, 227.073 mmol, 3 equiv) at room temperature under an air atmosphere. The resulting mixture was stirred overnight at room temperature under an air atmosphere. The reaction was quenched at room temperature by the addition of water (100 mL). The resulting mixture was extracted with CH 2 Cl 2 (3 × 100 mL). The combined organic layers were dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica column chromatography eluting with PE / EA (1:1, v / v) to give 2-(3-{[(tert-butyldimethylsilyl)oxy]methyl}phenyl)acetonitrile I-11b (19.5 g, 98.54%) as a colorless oil. 1 H NMR (DMSO-d 6 , 400 MHz) δ 0.09 (6H, s), 0.92 (9H, s), 4.04 (2H, s), 4.73 (2H, s), 7.24 (2H, dd), 7.35 (2H, dd).

[0199] Step 3: To a stirred solution of nitrile I-11b (19.5 g, 74.59 mmol, 1 equiv) in THF (100 mL) was added dropwise sodium hydride (3.58 g, 149.17 mmol, 2 equiv) at 0 °C under an air atmosphere. The resulting mixture was stirred for 1 h at room temperature under an air atmosphere. Dimethyl carbonate (53.75 g, 596.70 mmol, 8 equiv) was added dropwise to the above mixture at room temperature. The resulting mixture was stirred for an additional 3 h at room temperature. The reaction was quenched at room temperature by the addition of water / ice (100 mL). The resulting mixture was extracted with ethyl acetate (3 × 100 mL). The combined organic layers were dried over anhydrous Na 2 SO 4It was dehydrated. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica column chromatography eluted with PE / EA (1:1, volume / volume) to obtain methyl 2-(3-{[(tert-butyldimethylsilyl)oxy]methyl}phenyl)-2-cyanoacetate (10.1 g, 42.39%) as a yellow oil. 1 H NMR (CD 3 OD, 400 MHz) δ 0.13 (6H, d), 0.97 (9H, s), 3.78 (3H, s), 4.79 (2H, d), 7.22 - 7.46 (3H, m), 7.50 (1H, dq).

[0200] Step 4: To a stirred mixture of methyl 2-(3-{[(tert-butyldimethylsilyl)oxy]methyl}phenyl)-2-cyanoacetate (5 g, 15.65 mmol, 1 equivalent) and CoCl 2 ·6H 2 O (11.17 g, 46.95 mmol, 3 equivalents) in methanol (100 mL), Boc 2 O (10.25 g, 46.95 mmol, 3 equivalents) was added at room temperature under an air atmosphere. NaBH 4 (4.74 g, 125.21 mmol, 8 equivalents) was added portionwise to the above mixture at 0 °C. The resulting mixture was stirred at room temperature for an additional 3 hours. The reaction was quenched at room temperature by the addition of water (100 mL). The resulting mixture was extracted with CH 2 Cl 2 (3 × 50 mL). The combined organic layers were dehydrated with anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica column chromatography eluted with PE / EA (1:1, volume / volume) to obtain methyl 3-[(tert-butoxycarbonyl)amino]-2-(3-{[(tert-butyldimethylsilyl)oxy]methyl}phenyl)propanoate I-11c (2282 mg, 34.42%) as a yellow oil. LCMS: m / z (ESI), [M + H - Boc] + = 324.15. 1 H NMR (CD 3OD, 400 MHz) δ 0.12 (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).

[0201] Step 5: To a stirred solution of ester I-11c (200 mg, 0.472 mmol, 1 equiv) in THF (5 mL, 61.71 mmol) was added TBAF (370.32 mg, 1.42 mmol, 3 equiv) at room temperature under an air atmosphere. The resulting mixture was stirred at room temperature for 3 h under an air atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was purified by preparative TLC using CH 2 Cl 2 / methanol (40:1, v / v) to give methyl 3-[(tert-butoxycarbonyl)amino]-2-[3-(hydroxymethyl)phenyl]propanoate (97 mg, 66.41%) as a white solid. LCMS: m / z (ESI), [M+H] + = 310.20. 1 H NMR (DMSO-d 6 , 400 MHz) δ 1.37 (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).

[0202] Step 6: To a stirred solution of methyl 3-[(tert-butoxycarbonyl)amino]-2-[3-(hydroxymethyl)phenyl]propanoate (300 mg, 0.97 mmol, 1 equiv) in DCM (6 mL, 94.38 mmol, 97.33 equiv) were added N-methylcarbamoyl chloride (108.82 mg, 1.16 mmol, 1.2 equiv) and Et 3N (294.39 mg, 2.91 mmol, 3 equivalents) was added dropwise. The resulting mixture was stirred overnight at room temperature under an air atmosphere. The reaction was quenched at room temperature by the addition of water (10 mL). The resulting mixture was extracted with CH 2 Cl 2 (3 × 10 mL). The combined organic layers were dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, MeCN in water, 100% to 100% concentration gradient over 10 minutes; detector, UV 254 nm to give methyl 3-[(tert-butoxycarbonyl)amino]-2-(3-{[(methylcarbamoyl)oxy]methyl}phenyl)propanoate I-11d (90 mg, 25.33%) as a colorless oil. LCMS: m / z (ESI), [M+H] + = 367.15.

[0201]

[0203] Step 7: To a stirred solution of ester I-11d (270 mg, 0.74 mmol, 1 equivalent) in methanol (8 mL) and H 2 O (2 mL), LiOH (70.59 mg, 2.95 mmol, 4 equivalents) was added at room temperature under an air atmosphere. The resulting mixture was stirred at room temperature for 2 hours under an air atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, MeCN in water, 50% to 60% concentration gradient over 10 minutes; detector, UV 254 nm to give 3-[(tert-butoxycarbonyl)amino]-2-(3-{[(methylcarbamoyl)oxy]methyl}phenyl)propanoic acid I-11 (90 mg, 34.66%) as a white solid. LCMS: m / z (ESI), [M+H] + = 353.20.

[0202] Synthesis of Intermediate I-12

[0203]

Chemical Formula

[0204]

[0204] Step 1: To a stirred mixture of ethyl 2-cyano-2-(4-nitrophenyl)acetate I-12a (2.0 g, 8.539 mmol, 1 equiv) and zinc (4.47 g, 68.31 mmol, 8 equiv) in THF (40 mL), NH 4 Cl (4.57 g, 85.39 mmol, 10 equiv) in water (10 mL) was added dropwise at 25 °C under a nitrogen atmosphere. The resulting mixture was stirred at room temperature for 48 h under a nitrogen atmosphere. After the reaction, the resulting mixture was filtered and the filter cake was washed with methanol (3 × 20 mL). The filtrate was concentrated under reduced pressure. The residue was purified by silica column chromatography eluting with CH 2 Cl 2 / methanol (60:1, v / v) to give ethyl 2-(4-aminophenyl)-2-cyanoacetate I-12b (1.2 g, 68.81%) as a yellow solid. LCMS: m / z (ESI), [M+H] + = 205.15.

[0205]

[0205] Step 2: To a stirred mixture of nitrile I-12b (1.1 g, 5.39 mmol, 1 equiv) and triethylamine (1.64 g, 16.16 mmol, 3 equiv) in DCM (30 mL), triethylamine (1.64 g, 16.16 mmol, 3 equiv) was added dropwise at 0 °C under a nitrogen atmosphere. The resulting mixture was stirred at room temperature for 2 h under a nitrogen atmosphere. After the reaction, the resulting mixture was diluted with water (80 mL). The resulting mixture was extracted with ethyl acetate (3 × 60 mL). The combined organic layers were washed with saturated brine (3 × 100 mL) and dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure. The residue was dissolved in DCM (5 mL). The residue was purified by preparative TLC using 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-d 6, 400 MHz) δ 1.18 (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).

[0206] Step 3: To a stirred mixture of ester I-12c (1.1 g, 4.47 mmol, 1 equiv) and CoCl 2 ·6H 2 O (2.13 g, 8.93 mmol, 2 equiv) in methanol (40 mL), (Boc) 2 O (1.95 g, 8.93 mmol, 2 equiv) and NaBH 4 (0.68 g, 17.87 mmol, 4 equiv) were added portionwise at 0 °C under an air atmosphere. The resulting mixture was stirred overnight at room temperature under an air atmosphere. After the reaction, the resulting mixture was filtered and the filter cake was washed with water (3 × 30 mL). The resulting mixture was diluted with water (50 mL). The resulting mixture was extracted with ethyl acetate (3 × 40 mL). The combined organic layers were dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC using CH 2 Cl 2 / methanol (5:1, v / v) to give ethyl 3-[(tert-butoxycarbonyl)amino]-2-(4-acetamidophenyl)propanoate I-12d (900 mg, 57.50%) as a yellow oil. LCMS: m / z (ESI), [M - Boc] + = 251.15.

[0206]

[0207] Step 4. To a stirred mixture of ester I-12d (500 mg, 2.03 mmol, 1 equiv) in methanol (4.0 mL) was added dropwise LiOH (194.51 mg, 8.12 mmol, 4 equiv) in water (1.0 mL) at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at 60 °C overnight 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 reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, MeCN in water, gradient from 10% to 50% concentration in 30 min; detector, UV 254 nm and held at 16% for 10 min. After lyophilization, 2-(4-acetamidophenyl)-3-((tert-butoxycarbonyl)amino)propanoic acid I-12 (260 mg, 36.54%) was obtained as a white solid. LCMS: m / z (ESI), [M-t-Bu] + = 267.05.

[0207] Synthesis of Intermediate I-13

[0208]

Chem.

[0209]

[0208] Step 1: To a mixture of tert-butyl N-[2-(3-bromophenyl)-2-hydroxyethyl]carbamate I-13a (2.0 g, 6.33 mmol, 1 equiv) and (methylphosphonoyl)methane (0.59 g, 7.59 mmol, 1.2 equiv) in dioxane (25 mL) was added Pd(OAc) 2 (0.28 g, 1.27 mmol, 0.2 equiv), K 3 PO 4 (2.69 g, 12.65 mmol, 2.0 equiv) and Xantphos (1.10 g, 1.90 mmol, 0.3 equiv) portionwise, and 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 × 30 mL). The combined organic layers were dried over anhydrous Na 2 SO 4It was dehydrated. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, MeCN in water (0.1% FA), concentration gradient from 0% to 100% in 10 minutes; detector, UV254 nm. Thus, tert-butyl N-{2-[3-(dimethylphosphoryl)phenyl]-2-hydroxyethyl}carbamate I-13b (1.3 g, 65.59%) was obtained as a yellow oil. 1 H NMR (400 MHz, DMSO-d 6 ) δ 1.35 (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).

[0209] Step 2: A mixture of phthalimide (169.05 mg, 1.15 mmol, 1.2 eq), PPh 3 (753.39 mg, 2.87 mmol, 3 eq) in THF (1 mL) was treated with DIAD (580.81 mg, 2.87 mmol, 3 eq) at 0 °C for 0.5 h under a nitrogen atmosphere, and then alcohol I-13b (300 mg, 0.96 mmol, 1 eq) in THF (2 mL) was added dropwise at 0 °C for 1 h. The reaction was quenched with water at room temperature. The resulting mixture was extracted with CH 2 Cl 2 (3 × 20 mL), dehydrated with anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC using CH 2 Cl 2 / methanol (10:1, v / v) to obtain tert-butyl N-{2-[3-(dimethylphosphoryl)phenyl]-2-(1,3-dioxoisoindol-2-yl)ethyl}carbamate I-13c (270 mg, 63.74%) as a white solid. 1 H NMR (400 MHz, DMSO-d 6) δ 1.27 (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).

[0210] Step 3: A solution of phthalimide I-13c (240 mg, 0.54 mmol, 1 equiv) and hydrazine (216.97 mg, 4.34 mmol, 8 equiv) in methanol (5 mL) was stirred at room temperature for 4 h under a nitrogen atmosphere. The resulting mixture was filtered and the filter cake was washed with methanol (3 × 5 mL). The filtrate was concentrated under reduced pressure. The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, MeCN in water, 10% to 50% concentration gradient over 10 min; detector, UV 254 nm. This gave tert-butyl N-{2-amino-2-[3-(dimethylphosphoryl)phenyl]ethyl}-carbamate I-13 (82 mg, 48.40%) as a colorless oil. 1 H NMR (400 MHz, DMSO-d 6 ) δ 1.35 (9H, s), 1.63 (6H, d), 3.09 (3H, m), 6.84 (1H, t), 7.54 (4H, m). Synthesis of Intermediate I-14

[0210]

Chemical Structure

[0211]

[0211] Step 1: To a stirred solution of ester I-26c (2.08 g, 5.81 mmol, 1 equiv) and Pd(PPh 3 ) 4 (67.10 mg, 0.058 mmol, 0.01 equiv) in DMF (10 mL) was added Zn(CN) 2(688.59 mg, 5.86 mmol, 1.01 eq) was added portionwise. The resulting mixture was stirred at 80 °C for 2.0 h under a nitrogen atmosphere. After cooling to room temperature, the resulting mixture was diluted with water (60 mL). The resulting mixture was extracted with ethyl acetate (3 × 40 mL). The combined organic layers were washed with water (3 × 10 mL) and dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC using PE / EA (2:1, v / v) to give methyl 3-[(tert-butoxycarbonyl)amino]-2-(4-cyanophenyl)propanoate I-14c (1.3 g, 73.57%) as a yellow solid.

[0212]

[0212] Step 2: To a stirred solution of nitrile I-14c (600 mg, 1.97 mmol, 1 eq) in methanol (8.0 mL) and LiOH·H 2 O (330.89 mg, 7.88 mmol, 4.0 eq) in water (2.0 mL) was added dropwise at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at 70 °C for 2.0 h under a nitrogen atmosphere. After cooling to room temperature, the resulting mixture was concentrated under vacuum. After evaporation, the residue was dissolved in methanol (2.0 mL) and basified to pH 9 with an aqueous NH 3 solution. The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, MeCN in water, gradient from 10% to 50% concentration in 10 min; detector, UV 254 nm, held at 18.2% for 20 min to give 3-[(tert-butoxycarbonyl)amino]-2-(4-cyanophenyl)propanoic acid I-14 (400 mg, 69.89%) as a white solid. LCMS: m / z (ESI), [M- t Bu] + = 235.05.

[0213] Synthesis of Intermediate I-15

[0214]

Chemical Structure

[0215]

[0213] Step 1: In a 250 mL round-bottom flask, at 0 °C, (5-bromopyridin-2-yl)methanol I-15a (5 g, 26.59 mmol, 1 equiv), TBSCl (4.81 g, 31.91 mmol, 1.2 equiv), triethylamine (8.07 g, 79.78 mmol, 3 equiv), DMAP (0.32 g, 2.66 mmol, 0.1 equiv) and DCM (80 mL) were added. The resulting mixture was stirred at room temperature overnight. The resulting mixture was extracted with CH 2 Cl 2 (3 × 30 mL), and dehydrated with anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica column chromatography eluting with EA / CH 2 Cl 2 (3:1, v / v) 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 (CD 3 OD, 400 MHz) δ 0.15 (6H, s), 0.93 (9H, d), 4.77 (2H, d), 7.50 (1H, m), 8.01 (1H, m), 8.55 (1H, m).

[0214] Step 2: In six 500 mL round-bottom flasks, at room temperature, 5-bromo-2-{[(tert-butyldimethylsilyl)oxy]methyl}pyridine (6.3 g, 20.84 mmol, 1 equiv), methyl cyanoacetate (2.48 g, 25.01 mmol, 1.2 equiv), Pd(OAc) 2 (0.47 g, 2.08 mmol, 0.1 equiv), dppf (2.30 g, 4.17 mmol, 0.2 equiv), t-BuOK (7.02 g, 62.52 mmol, 3 equiv) and 1,4-dioxane (240 mL) were added. The resulting mixture was stirred at 70 °C for 5 h under a nitrogen atmosphere. The mixture was quenched with saturated brine (200 mL), extracted with EA (2 × 200 mL), and the combined organic layers were dried over anhydrous Na 2 SO 4It was dehydrated, filtered, and evaporated to obtain a crude solid. The residue was purified by silica column chromatography eluting with PE / EA (1:1, v / v) to give 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 (CD 3 OD, 400 MHz) δ 0.17 (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).

[0215] Step 3: In a 50 mL round-bottom flask, at room temperature, nitrile I-15b (2.24 g, 6.99 mmol, 1 equiv), CoCl 2 ·6H 2 O (2.49 g, 10.49 mmol, 1.5 equiv), Boc 2 O (3.05 g, 13.98 mmol, 2.0 equiv) and methanol (20 mL) were added, and NaBH 4 (1.06 g, 27.96 mmol, 4.0 equiv) was added portionwise at 0 °C. The resulting mixture was stirred at room temperature overnight. The reaction was quenched with water (100 mL) at room temperature. The resulting mixture was filtered, and the filtered cake was washed with CH 2 Cl 2 (3 × 30 mL). The filtrate was extracted with DCM (2 × 100 mL), and the combined organic layers were dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC using CH 2 Cl 2 / methanol (17:1, v / v) to give methyl 3-[(tert-butoxycarbonyl)amino]-2-(6-{[(tert-butyldimethylsilyl)oxy]methyl}pyridin-3-yl)propanoate (625 mg, 14.74%) as a yellowish-brown solid. LCMS: m / z (ESI), [M+H] + = 425.30.

[0216]

[0216] Step 4: In a 50 mL round-bottom flask, at room temperature, methyl 3-[(tert-butoxycarbonyl)amino]-2-(6-{[(tert-butyldimethylsilyl)oxy]methyl}pyridin-3-yl)propanoate (600 mg, 1.41 mmol, 1 equiv), LiOH (67.69 mg, 2.83 mmol, 2 equiv), water (5 mL) and THF (5 mL) were added. The resulting mixture was stirred at room temperature overnight. The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, methanol in water, 0 to 10% concentration gradient in 10 minutes; detector, UV 254 nm to obtain 3-[(tert-butoxycarbonyl)amino]-2-[6-(hydroxymethyl)pyridin-3-yl]propanoic acid I-15 (300 mg, 63.77%) as a light brown solid. LCMS: m / z (ESI), [M+H] + = 297.10。

[0217] Synthesis of Intermediate I-16

[0218]

Chemical formula

[0219]

[0217] Step 1: A mixture of 3-bromo-5-methylpyridine I-16a (5 g, 29.07 mmol, 1 equiv) and methyl 2-cyanoacetate (3.46 g, 34.88 mmol, 1.2 equiv), Pd(OAc) 2 (0.65 g, 2.91 mmol, 0.1 equiv), dppf (3.21 g, 5.81 mmol, 0.2 equiv), potassium 2-methylpropane-2-olate (9.78 g, 87.20 mmol, 3 equiv) in 1,4-dioxane (100 mL) was stirred at 70 °C for 2 hours under a nitrogen atmosphere. The reaction was monitored by LCMS. The mixture was acidified to pH 9 using saturated NH 4 Cl (aqueous solution). The resulting mixture was stirred at room temperature for 1 hour. The resulting mixture was extracted with ethyl acetate (3 × 300 mL). The combined organic layers were dried over anhydrous Na 2 SO4 It was dehydrated. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica column chromatography eluting with CH 2 Cl 2 / methanol (30:1, v / v) 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.

[0220]

[0218] Step 2: To a stirred mixture of nitrile I-16b (800 mg, 4.21 mmol, 1 equiv) and Boc 2 O (3671.87 mg, 16.82 mmol, 4 equiv), CoCl 2 ·6H 2 O (2601.83 mg, 10.94 mmol, 2.6 equiv) in methanol (50 mL) was added NaBH 4 (1272.92 mg, 33.65 mmol, 8 equiv) portionwise at 0 °C under a nitrogen atmosphere. The resulting mixture was stirred at room temperature for 1 h under a nitrogen atmosphere. The reaction was quenched with water at room temperature. The resulting mixture was extracted with CH 2 Cl 2 (2 × 100 mL). The combined organic layers were dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC using CH 2 Cl 2 / methanol (32:1, v / v) to give methyl 3-[(tert-butoxycarbonyl)amino]-2-(5-methylpyridin-3-yl)propanoate I-16c (310 mg, 43.66%) as a yellow solid. LCMS: m / z (ESI), [M+H] + = 295.15.

[0221]

[0219] Step 3: Ester I-16c (300 mg, 1.02 mmol, 1 equiv) in methanol (3 mL) and LiOH·H 2 O in H 2A solution of O (85.53 mg, 2.04 mmol, 2 equiv) was stirred at room temperature for 1 h under a nitrogen atmosphere. The residue was acidified to pH 5 with HCl (1 M). The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, methanol in water, concentration gradient from 10% to 23% in 30 min; detector, UV 220 nm to give 3-[(tert-butoxycarbonyl)amino]-2-(5-methylpyridin-3-yl)propanoic acid I-16 (140 mg, 49.00%) as a yellow solid. LCMS: m / z (ESI), [M+H] + = 281.10.

[0222] Synthesis of Intermediate I-17

[0223]

Chem.

[0224]

[0220] Step 1: In a 250 mL round-bottom flask, bromide I-17a (2.26 g, 22.80 mmol, 1.2 equiv), dppf (1.05 g, 1.90 mmol, 0.1 equiv), t-BuOK (6.40 g, 57.01 mmol, 3.0 equiv), palladium acetate (0.21 g, 0.95 mmol, 0.05 equiv) and methyl 2-cyano-2-methylacetate (2.58 g, 22.80 mmol, 1.2 equiv) in dioxane (70 mL) were added at room temperature. The resulting mixture was stirred at 70 °C for 3 h under a nitrogen atmosphere. The residue was quenched with saturated NH 4 Cl (100 mL). The resulting mixture was filtered and the filtrate was extracted with ethyl acetate (2 × 150 mL). The combined organic layers were dried over anhydrous Na 2 SO 4 and filtered and concentrated under reduced pressure. The residue was purified by silica column chromatography eluting with PE / EA (10:1, v / v) to give methyl 2-[3-(benzyloxy)phenyl]-2-cyanoacetate I-17b (1.8 g, 33.67%) as a yellow oil. 1 1H NMR (DMSO-d 6, 400 MHz) δ 3.71 (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).

[0221] Step 2: In a 40 mL vial, at 0 °C, nitrile I-17b (2 g, 7.11 mmol, 1 equivalent) in methanol (5 mL), CoCl 2 ·6H 2 O (4.40 g, 18.49 mmol, 2.6 equivalents) and Boc 2 O (6.21 g, 28.44 mmol, 4.0 equivalents) were added. NaBH 4 (2.15 g, 56.88 mmol, 8.0 equivalents) was added to the above mixture at 0 °C. The resulting mixture was stirred at room temperature for 2 hours. The reaction was quenched with water at 0 °C. The precipitated solid was filtered off and washed with DCM (1 × 100 mL). The resulting mixture was extracted with DCM (3 × 100 mL). The combined organic layers were dried over anhydrous Na 2 SO 4 and filtered, and concentrated under reduced pressure. The residue was purified by preparative TLC using CH 2 Cl 2 / methanol (10:1, volume / volume) to give methyl 2-[3-(benzyloxy)phenyl]-3-[(tert-butoxycarbonyl)amino]propanoate I-17c (1.1 g, 40.14%) as a yellow solid. LCMS: m / z (ESI), [M + H - t Bu] + = 330.10. 1 1H NMR (400 MHz, DMSO-d 6 ) δ 1.35 (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).

[0222] Step 3: In a 50 mL round-bottom flask, add ester I-17c (1.1 g, 2.85 mmol, 1 equivalent) in THF (2 mL) at room temperature and LiOH·H 2 O (239.49 mg, 5.71 mmol, 2 equivalents) in water (2 mL). Stir the resulting mixture at room temperature for 1.5 h. Acidify the mixture to pH 5 with HCl (aqueous solution). Purify the residue by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, methanol in water, concentration gradient from 0% to 100% in 30 min; detector, UV 254 nm. Concentrate the resulting mixture under vacuum to obtain 2-[3-(benzyloxy)phenyl]-3-[(tert-butoxycarbonyl)amino]propanoic acid I-17 (500 mg, 47.17%) as a yellow solid. LCMS: m / z (ESI), [M+H- t Bu] + = 316.10. 1 H NMR (400 MHz, DMSO-d 6 ) δ 1.34 (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). Synthesis of Intermediate I-18

[0225]

Chemical Structure

[0226]

[0223] Step 1: Methyl 2-(pyridin-3-yl)acetate I-18a (4 g, 26.46 mmol, 1 equivalent) and K 2 CO 3(10.97 g, 79.38 mmol, 3 equiv) and a mixture of formaldehyde (1.59 g, 52.92 mmol, 2 equiv) in DMF (40 mL) were stirred at 80 °C for 2 h under a nitrogen atmosphere. 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 × 50 mL). The combined organic layers were washed with saturated sodium chloride solution (3 × 50 mL) and dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica column chromatography eluting with PE / EA (2:3, v / v) 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 (CDCl 3 , 400 MHz) δ 3.87 (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).

[0224] Step 2: To a stirred mixture of enolate I-18b (757 mg, 4.64 mmol, 1 equiv) in DMF (10 mL) was added 1-methylpiperazine (2323.41 mg, 23.20 mmol, 5 equiv) at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at room temperature for 2 h under a nitrogen atmosphere. The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, MeCN in water, 10% to 50% concentration gradient over 10 min; detector, UV254 nm. The resulting mixture was concentrated under vacuum. This gave methyl 3-(4-methylpiperazin-1-yl)-2-(pyridin-3-yl)propanoate I-18c (884 mg, 72.36%) as a yellow solid. LCMS: m / z (ESI), [M+H] + = 264.30.

[0227]

[0225] Step 3: Piperazine I-18c (150 mg, 0.57 mmol, 1 equiv) in THF (8 mL) and H2 To the stirred mixture in O(2 mL), LiOH (68.2 mg, 2.85 mmol, 5 eq) was added portionwise at room temperature under a nitrogen atmosphere. The resulting mixture was stirred overnight at room temperature under a nitrogen atmosphere. The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, MeCN in water, concentration gradient from 10% to 50% in 10 min; detector, UV 254 nm. This gave 3-(4-methylpiperazin-1-yl)-2-(pyridin-3-yl)propanoic acid I-18 (120 mg, 84.5%) as a white solid. LCMS: m / z (ESI), [M+H] + = 250.35

[0228] Synthesis of Intermediate I-19

[0229]

Chemical Structure

[0230]

[0226] Step 1: In a 500 mL round-bottom flask, tert-butyl 3-bromobenzoate I-19a (8 g, 31.11 mmol, 1 eq) and methyl cyanoacetate (4624.44 mg, 46.67 mmol, 1.5 eq), Pd(OAc) 2 (698.52 mg, 3.11 mmol, 0.1 eq), dppf (3437.21 mg, 6.22 mmol, 0.2 eq), t-BuOK (10.47 g, 93.34 mmol, 3 eq) were added at room temperature in dioxane (200 mL). The resulting mixture was stirred at 70 °C for 6 h under a nitrogen atmosphere. The mixture was acidified to pH 6 with saturated NH 4 Cl (aqueous solution). The resulting mixture was extracted with ethyl acetate (3 × 50 mL). The combined organic layers were dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica column chromatography eluting with PE / EA (7:1, v / v) to give tert-butyl 3-(1-cyano-2-methoxy-2-oxoethyl)benzoate I-19b (3.7 g, 43.20%) as a pale yellow oil. 11H NMR (CD 3 OD, 400 MHz) δ 1.63 (9H, s), 3.81 (3H, s), 7.56 (1H, td), 7.70 (1H, ddd), 8.02 (1H, dt), 8.05 - 8.09 (1H, m).

[0227] Step 2: In a 100 mL round-bottom flask, at room temperature, nitrile I-19b (3.7 g, 13.44 mmol, 1 equivalent) and CoCl 2 ·6H 2 O (2.62 g, 20.16 mmol, 1.5 equivalents), Boc 2 O (5.87 g, 26.88 mmol, 2 equivalents) were added in methanol (20 mL). NaBH 4 (1.53 g, 40.32 mmol, 3 equivalents) was added portionwise to the above mixture at 0 °C over 30 minutes. The resulting mixture was stirred at room temperature for an additional 15 hours. The reaction was quenched with water at room temperature. The resulting mixture was extracted with DCM (3 × 40 mL). The combined organic layers were dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica column chromatography eluting with PE / EA (5:1, v / v) to give tert-butyl 3-{3-[(tert-butoxycarbonyl)amino]-1-methoxy-1-oxopropan-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.

[0231]

[0228] Step 3: In a 100 mL round-bottom flask, ester I-19c (2 g, 5.27 mmol, 1 equiv) and LiOH (252.47 mg, 10.54 mmol, 2 equiv) in water (15 mL) and methanol (15 mL) were added at room temperature. The resulting mixture was stirred at room temperature for 2 h under a nitrogen atmosphere. The mixture was acidified to pH 5 with HCl (1 M). The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, methanol in water, concentration gradient from 12% to 22% in 25 min; detector, UV 254 nm. Thereby, 3-[(tert-butoxycarbonyl)amino]-2-[3-(tert-butoxycarbonyl)phenyl]propanoic acid I-19 (650 mg, 33.75%) was obtained as an off-white solid. LCMS: m / z (ESI), [M+H] + = 366.00.

[0232] Synthesis of Intermediate I-20

[0233]

Chemical Structure

[0234]

[0229] Step 1. In a 40 mL vial, methyl 2-bromo-2-phenylacetate I-20a (1 g, 4.37 mmol, 1 equiv), tert-butyl N-[2-(piperazin-1-yl)ethyl]carbamate (1.20 g, 5.24 mmol, 1.2 equiv), and triethylamine (1.33 g, 13.10 mmol, 3 equiv) in THF (20 mL) were added at room temperature. The resulting mixture was stirred at 70 °C for 15 h under a nitrogen atmosphere. The reaction was quenched at room temperature by the addition of water (60 mL). The resulting mixture was extracted with EA (3 × 50 mL). The combined organic layers were washed with saturated brine (1 × 100 mL), anhydrous Na 2 SO 4It was dehydrated. After filtration, the filtrate was concentrated under reduced pressure. The residue was dissolved in DCM (4 mL). The residue was purified by preparative TLC using 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%) as a pale yellow oil. LCMS: m / z (ESI), [M+H] + = 378.10.

[0235]

[0230] In a 40 mL vial, piperazine I-20b (600 mg, 1.59 mmol, 1 equiv) and LiOH (76.14 mg, 3.18 mmol, 2 equiv) in THF (5 mL) and water (5 mL) were added at room temperature. The resulting mixture was stirred at room temperature for 15 h under a nitrogen atmosphere. The mixture was acidified to pH 5 with HCl (aqueous solution). The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, methanol in water, concentration gradient from 5% to 10% in 30 min; detector, UV 220 nm. The resulting mixture was concentrated under reduced pressure. Thereby, (4-{2-[(tert-butoxycarbonyl)amino]ethyl}piperazin-1-yl)(phenyl)acetic acid I-20 (469 mg, 81.18%) was obtained as an off-white solid. LCMS: m / z (ESI), [M+H] + = 364.10. 1 H NMR (CD 3 OD, 400 MHz) δ 1.44 (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). Synthesis of Intermediate I-21

[0236]

Chemical Structure

[0237]

[0231] Step 1: A mixture of 3-(cyanomethyl)benzoic acid I-21a (5 g, 31.03 mmol, 1 equiv) and CDI (5.53 g, 34.13 mmol, 1.1 equiv) in THF (26 mL) was stirred at room temperature for 3 h. H 2 O (20 mL) of NaBH 4 (3.52 g, 93.08 mmol, 3 equiv) was added dropwise to the above mixture at 0 °C under a nitrogen atmosphere. The resulting mixture was stirred at room temperature overnight under a nitrogen atmosphere. The reaction was monitored by TLC using PE / EA (6:1, v / v). The reaction was quenched with water at room temperature. The resulting mixture was extracted with DCM (3 × 200 mL), and the combined organic layers were dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica column chromatography eluting with PE / EA (8:1, v / v) to afford 2-[3-(hydroxymethyl)phenyl]acetonitrile (2.3 g, 50.37%) as a pale yellow liquid. 1 H NMR (CD 3 OD, 400 MHz) δ 3.91 (2H, s), 4.64 (2H, s), 7.00 - 7.07 (1H, m), 7.17 (1H, t), 7.39 (1H, q), 7.99 (1H, s).

[0232] Step 2: To a stirred mixture of 2-[3-(hydroxymethyl)phenyl]acetonitrile (2.3 g, 15.63 mmol, 1 equiv), triethylamine (4.74 g, 46.88 mmol, 3 equiv), and DMAP (0.02 g, 0.16 mmol, 0.01 equiv) in DCM (10 mL) was added TBSCl (2.83 g, 18.75 mmol, 1.2 equiv) portionwise at 0 °C under a nitrogen atmosphere. The resulting mixture was stirred at room temperature overnight under a nitrogen atmosphere. The reaction was monitored by TLC using PE / EA (6:1, v / v). The reaction was quenched with water at room temperature. The resulting mixture was extracted with CH 2 Cl 2 (3 × 100 mL), and the combined organic layers were dried over anhydrous Na 2 SO 4It was dehydrated. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC using PE / EA (6:1, volume / volume) 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 (CD 3 OD, 400 MHz) δ 0.13 (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).

[0233] Step 3. To a stirred mixture of nitrile I-21b (2.1 g, 8.03 mmol, 1 equivalent) in THF (25 mL), sodium hydride (289.14 mg, 12.05 mmol, 1.5 equivalents) was added portionwise at 0 °C under a nitrogen atmosphere. The resulting mixture was stirred at room temperature for 1 hour under a nitrogen atmosphere. Next, dimethyl carbonate (2.89 g, 32.13 mmol, 4 equivalents) was added portionwise at 0 °C under a nitrogen atmosphere. The final reaction mixture was stirred at room temperature overnight. The reaction was monitored by TLC using PE / EA (6:1, volume / volume). The reaction was quenched with water at room temperature. The resulting mixture was extracted with ethyl acetate (3 × 150 mL), and the combined organic layers were dried over anhydrous Na 2 SO 4 and dehydrated. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC using PE / EA (6:1, volume / volume) 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 (CD 3 OD, 400 MHz) δ 0.13 (6H, s), 0.97 (9H, s), 3.78 (3H, s), 4.80 (2H, q), 7.32 - 7.51 (4H, m).

[0234] Step 4. Methyl 2-(3-{[(tert-butyldimethylsilyl)oxy]methyl}phenyl)-2-cyanoacetate (1.9 g, 5.95 mmol, 1 equivalent) and (Boc)2 O (5.19 g, 23.79 mmol, 4 equivalents) and CoCl 2 ·6H 2 O (8.9 g, 8.96 mmol, 1.5 equivalents) was added portionwise to the stirred mixture in methanol (50 mL) at 0 °C under a nitrogen atmosphere with NaBH 4 (1.80 g, 47.58 mmol, 8 equivalents). The resulting mixture was stirred overnight at room temperature under a nitrogen atmosphere. The resulting mixture was filtered. The filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC using PE / EA (5:1, volume / volume) to give methyl 3-[(tert-butoxycarbonyl)amino]-2-(3-{[(tert-butyldimethylsilyl)oxy]methyl}phenyl)propanoate (1.2 g, 47.63%) as a pale yellow liquid. LCMS: m / z (ESI), [M+H] + = 324.20.

[0238]

[0235] Step 5. Methyl 3-[(tert-butoxycarbonyl)amino]-2-(3-{[(tert-butyldimethylsilyl)oxy]methyl}phenyl)propanoate (1.2 g, 2.83 mmol, 1 equivalent) in methanol (1 mL) and H 2 O (1 mL) of LiOH·H 2 O (237.72 mg, 5.67 mmol, 2 equivalents) was stirred overnight at 60 °C under a nitrogen atmosphere. The mixture was acidified to pH 5 with HCl (1 M). The resulting mixture was concentrated under vacuum. The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, methanol in water, 0% to 5% concentration gradient over 30 minutes; detector, UV220 nm. This gave 3-[(tert-butoxycarbonyl)amino]-2-[3-(hydroxymethyl)phenyl]propanoic acid I-21 (530 mg, 63.35%) as an off-white solid. LCMS: m / z (ESI), [M- t Bu] + = 240.06.

[0239] Synthesis of Intermediate I-22

[0240]

Chemical Structure

[0241]

[0236] Step 1: To a solution of ethyl cyanoacetate (10 g, 88.41 mmol, 1.00 equiv) in DMF (50 ml), sodium hydride (2.55 g, 106.09 mmol, 1.2 equiv) was added portionwise, and the resulting mixture was stirred at 0 °C for 1 h under a nitrogen atmosphere. Subsequently, 4-fluoronitrobenzene I-22a (8.73 g, 61.87 mmol, 0.70 equiv) in dioxane (50 ml) was added dropwise at 0 °C. The resulting mixture was stirred at 80 °C for 12 h. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in DCM (300 mL). The resulting mixture was washed with 2 × 250 mL of HCl (1 M), 2 × 250 ml of water, and 1 × 250 mL of saturated brine. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica column chromatography eluting with PE / EA (8:1, v / v) 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 (400 MHz, DMSO-d 6 ) δ 1.20 (3H, t), 4.15 (2H, m), 5.97 (1H, s), 7.74 (2H, d), 8.29 (2H, d).

[0237] Step 2: A solution of nitrile I-22b (1 g, 4.54 mmol, 1 equiv) in methanol was treated with Boc 2 O (3.96 g, 18.17 mmol, 4 equiv) and CoCl 2 ·6H 2 O (2.81 g, 11.81 mmol, 2.6 equiv) at 0 °C for 5 min under a nitrogen atmosphere, and then NaBH 4 (1.37 g, 36.34 mmol, 8 equiv) was added portionwise at 0 °C. The resulting mixture was stirred at room temperature for 2 h. The reaction was quenched with water at room temperature. The resulting mixture was filtered, and the filtered cake was washed with methanol (3 × 20 mL). The aqueous layer was extracted with CH 2 Cl2 (3 × 100 mL) was extracted. The resulting mixture was concentrated under reduced pressure. The residue was purified by preparative TLC using CH 2 Cl 2 / methanol (2:1, v / v) to give ethyl 3-((tert-butoxycarbonyl)amino)-2-(4-((tert-butoxycarbonyl)amino)phenyl)propanoate I-22c (500 mg, 27.91%) as a yellow oil. LCMS: m / z (ESI), [M + H] + = 409.25.

[0242]

[0238] Step 3: In a 100 mL round-bottom flask, at room temperature, ester I-22c (500 mg, 1.27 mmol, 1 equiv) and LiOH·H 2 O (106.37 mg, 2.54 mmol, 2 equiv) in THF (5 mL) and H 2 O (5 mL) were added. The resulting mixture was stirred at room temperature for 15 h under an air atmosphere. The mixture was acidified to pH 5 with HCl (1 M). The resulting mixture was concentrated under reduced pressure. The resulting oil was dried by lyophilization. This gave 3-[(tert-butoxycarbonyl)amino]-2-{4-[(tert-butoxycarbonyl)amino]phenyl}propanoic acid I-22 (420 mg, 68.74%) as a yellow solid. LCMS: m / z (ESI), [M - H] - = 379.15.

[0243] Synthesis of Intermediate I-23

[0244]

Chemical formula

[0245]

[0239] Step 1: A mixture of 2-(2-methoxyphenyl)acetonitrile I-23a (3 g, 20.38 mmol, 1 equiv) and sodium hydride (391.04 mg, 16.31 mmol, 1.2 equiv) in THF (30 mL) was stirred at room temperature for 2 h under a nitrogen atmosphere. Dimethyl carbonate (4.60 g, 50.96 mmol, 2.5 equiv) was added dropwise to the above mixture at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at room temperature for 2 h under a nitrogen atmosphere. The resulting mixture was quenched with water (40 mL). The aqueous layer was extracted with CH 2 Cl 2 (3 × 20 mL) and dried over anhydrous Na 2 SO 4 . The resulting mixture was concentrated under reduced pressure. The crude product was purified by reverse-phase flash 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 (400 MHz, DMSO-d 6 ) δ 3.72 (3H, s), 3.83 (3H, d), 7.01 (1H, t), 7.12 (1H, d), 7.34 (1H, d), 7.42 (1H, d).

[0240] Step 2: To a stirred mixture of methyl 2-cyano-2-(2-methoxyphenyl)acetate (1.9 g, 9.26 mmol, 1 equiv) and Boc 2 O (4.04 g, 18.52 mmol, 2 equiv) in methanol, CoCl 2 (3.01 g, 23.15 mmol, 2.5 equiv) and NaBH 4 (2.80 g, 74.07 mmol, 8 equiv) were added portionwise at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at room temperature for 2 h under a nitrogen atmosphere. The resulting mixture was quenched with water (30 mL). The aqueous layer was extracted with CH 2 Cl 2 (3 × 20 mL) and dried over anhydrous Na 2 SO 4It was dehydrated. The resulting mixture was concentrated under reduced pressure. The crude product was purified by reverse-phase flash to obtain methyl 3-[(tert-butoxycarbonyl)amino]-2-(2-methoxyphenyl)propanoate I-23b (1.3 g, 45.39%) as a yellow solid. LCMS: m / z (ESI), [M+H] + = 310.10. 1 H NMR (400 MHz, DMSO-d 6 ) δ 1.34 (9H, s), 3.20 (1H, d), 3.50 (1H, td), 3.56 (3H, s), 3.76 (3H, 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).

[0241] Step 3: A mixture of methyl 3-[(tert-butoxycarbonyl)amino]-2-(2-methoxyphenyl)propanoate (1.3 g, 4.20 mmol, 1 equiv) and LiOH (0.40 g, 16.81 mmol, 4 equiv) in methanol (4 mL) and water (1 mL) was stirred at room temperature for 2 h under a nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure. The crude product was purified by reverse-phase flash to obtain 3-[(tert-butoxycarbonyl)amino]-2-(2-methoxyphenyl)propanoic acid I-23 (800 mg, 64.46%) as a yellow solid. 1 H NMR (400 MHz, DMSO-d 6 ) 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). Synthesis of Intermediate I-24

[0246]

Chemical formula

[0247]

[0242] Step 1: A solution of 4-fluorobenzeneacetonitrile I-24a (3 g, 22.20 mmol, 1 equiv) in THF was treated with sodium hydride (1.1 g, 45.84 mmol, 2.06 equiv) at 0 °C for 1 h under a nitrogen atmosphere, followed by the addition of dimethyl carbonate (4.40 g, 48.84 mmol, 2.2 equiv) dropwise at room temperature. The resulting mixture was stirred at room temperature for 12 h. The reaction was quenched with water at room temperature. The resulting mixture was extracted with ethyl acetate (3 × 30 mL), and dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC using PE / EA (3:1, v / v) to afford methyl 2-cyano-2-(4-fluorophenyl)acetate (1.1 g, 15.01%) as a yellow oil. 1 1H NMR (DMSO-d 6 , 400 MHz) δ 3.74 (3H, s), 5.73 (1H, s), 7.25 - 7.35 (2H, m), 7.46 - 7.55 (2H, m).

[0243] Step 2: A mixture of methyl 2-cyano-2-(4-fluorophenyl)acetate (1 g, 5.18 mmol, 1 equiv), Boc 2 O (2.26 g, 10.36 mmol, 2.00 equiv) and CoCl 2 ·6H 2 O (2.46 g, 10.34 mmol, 2.00 equiv) in methanol was stirred at 0 °C for 5 min under a nitrogen atmosphere. Sodium borohydride (1.57 g, 41.42 mmol, 8 equiv) was added to the mixture at 0 °C to room temperature. The resulting mixture was filtered, and the filter cake was washed with CH 2 Cl 2 (3 × 30 mL). The filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC using PE / EA (5:1, v / v) to afford methyl 3-[(tert-butoxycarbonyl)amino]-2-(4-fluorophenyl)propanoate I-24b (400 mg, 25.68%) as a yellow oil. LCMS: m / z (ESI), [M+H] + = 298.25.

[0248]

[0244] Step 3. In a 50 mL round-bottom flask, methyl 3-[(tert-butoxycarbonyl)amino]-2-(4-fluorophenyl)propanoate I-24b (380 mg, 1.28 mmol, 1 equiv), LiOH (61.22 mg, 2.56 mmol, 2.00 equiv), and THF (2 mL) and water (1 mL) were added at room temperature. The resulting mixture was stirred at 60 °C for 8 h. The mixture was acidified to pH 4 - 6 with concentrated HCl. The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, methanol in water, concentration gradient from 10% to 50% in 10 min; detector, UV 254 nm. Thereby, 3-[(tert-butoxycarbonyl)amino]-2-(4-fluorophenyl)propanoic acid I-24 (260 mg, 63.98%) was obtained as a yellow solid. LCMS: m / z (ESI), [M - 56] + = 228.05.

[0249] Synthesis of Intermediate I-25

[0250]

Chem.

[0251]

[0245] 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 for 24 h using a Dean-Stark trap. Half of the toluene was evaporated under reduced pressure, the solution was cooled to 0 °C, and the product was recrystallized to obtain tert-butyl N-(benzotriazol-1-ylmethyl)carbamate I-25b (24 g, yield 58%) as a white solid. LCMS: 3-minute chromatography (3 min - water (6 mmol / L NH 4 HCO 3)In 5 - 95% MeCN, Waters Acquity UPLC BEH C18 1.7um, 2.1×50mm, 40 °C), t R = 1.55 minutes, MS(ESI) m / z = 249.1 [M+H] + . 1 H NMR: (400 MHz, DMSO - d 6 ) δ 8.42 (t, J = 6.4 Hz, 1H), 8.04 (d, J = 8.4 Hz, 1H), 7.96 (d, J = 8.4 Hz, 1H), 7.57 (t, J = 7.6 Hz, 1H), 7.44 - 7.38 (m, 1H), 5.88 (d, J = 6.7 Hz, 2H), 1.37 (s, 9H).

[0246] Step 2: To (R)-4 - benzyloxazolidin - 2 - one (9.1 g, 51.35 mmol) in THF (100 mL) at -78 °C, 2.5 M n - butyllithium (3.32 g, 51.87 mmol, 20.75 mL) was slowly added along the side of the flask, and the reaction mixture was stirred at -78 °C for 20 minutes. Next, a solution containing the lithium anion of (R)-4 - benzyloxazolidin - 2 - one in THF was cannulated into a pre - cooled (-78 °C) solution of 2-(3 - methoxyphenyl)acetyl chloride I - 25c (10.05 g, 54.44 mmol) in THF (100 mL). The reaction mixture was stirred at -78 °C for 1 hour and then poured into NH 4 Cl (saturated) / HCl (1 M) (3:1, volume / volume), and extracted with ethyl acetate. The combined extracts were dried over anhydrous sodium sulfate, filtered, and evaporated under reduced pressure. The residue was purified by silica flash chromatography eluting with a concentration gradient of 0 to 40% ethyl acetate in petroleum ether. The pure fractions were lyophilized to obtain (4R)-4 - benzyl - 3-[2-(3 - methoxyphenyl)acetyl]oxazolidin - 2 - one I - 25d (12.19 g, yield 73%) as a white solid. LCMS: 3 - minute chromatography (3 minutes - water (6 mmol / L NH 4 HCO 3) 5 - 95% MeCN, Waters Acquity UPLC BEH C18 1.7um, 2.1×50mm, 40°C), t R = 1.82 min, MS(ESI) m / z = 326.2 [M+H] + . 1 H NMR: (400 MHz, DMSO-d 6 ) δ 7.32 - 7.20 (m, 4H), 7.19 - 7.08 (m, 2H), 6.86 (d, J = 7.1 Hz, 3H), 4.68 (ddd, J = 10.8, 7.6, 2.9 Hz, 1H), 4.34 (t, J = 8.5 Hz, 1H), 4.25 (d, J = 15.9 Hz, 1H), 4.20 (dd, J = 8.9, 2.8 Hz, 1H), 4.10 (d, J = 15.9 Hz, 1H), 3.75 (s, 3H), 2.95 (ddd, J = 20.9, 13.5, 5.3 Hz, 2H).

[0247] Step 3: To a solution of 1M LiHMDS (10.68 g, 44.59 mmol) in THF (44.59 mL) cooled to -70°C was added via cannula a cooled (-70°C) solution of (4R)-4-benzyl-3-[2-(3-methoxyphenyl)acetyl]oxazolidin-2-one I-25d (12.19 g, 37.47 mmol) in THF (60 mL). The mixture was stirred at -70°C for 40 minutes. Then a solution of tert-butyl N-(benzotriazol-1-ylmethyl)carbamate I-25b (11.16 g, 44.96 mmol) in THF (60 mL), also cooled to -70°C, was added via cannula. The solution was warmed from -25 to -30°C and stirred for 2 hours. The mixture was extracted with ethyl acetate and NH 4It was poured into Cl(saturated) / HCl(1M)(3:1), and the aqueous phase was further extracted once more with ethyl acetate. The combined organic matter was dried over anhydrous sodium sulfate, filtered, and evaporated under reduced pressure. The residue was purified by a silica flash chromatography column eluting with a concentration gradient of 0 to 4% ethyl acetate in DCM. The pure fraction was lyophilized to obtain tert-butyl N-[(2S)-3-[(4R)-4-benzyl-2-oxo-oxazolidin-3-yl]-2-(3-methoxyphenyl)-3-oxo-propyl]carbamate I-25e (15 g, yield 88%) as a soft white solid. LCMS: 3-minute chromatography (3 minutes - 5 to 95% MeCN in water (0.02% TFA), Waters Acquity UPLC BEH C18 1.7um, 2.1×50mm, 40 °C) with t R = 1.95 minutes, MS(ESI) m / z = 455.3 [M+H] + . 1 H NMR: (500 MHz, DMSO-d 6 ) δ 7.37 - 7.17 (m, 6H), 7.08 (t, J = 5.6 Hz, 1H), 6.83 (t, J = 7.1 Hz, 2H), 6.78 (s, 1H), 5.16 (dd, J = 8.4, 6.1 Hz, 1H), 4.68 (t, J = 7.6 Hz, 1H), 4.25 (t, J = 8.5 Hz, 1H), 4.17 (dd, J = 12.4, 10.3 Hz, 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, 4.9 Hz, 2H), 1.35 (s, 9H).

[0248] Step 4: To tert-butyl N-[(2S)-3-[(4R)-4-benzyl-2-oxo-oxazolidin-3-yl]-2-(3-methoxyphenyl)-3-oxo-propyl]carbamate I-25e (15 g, 33.00 mmol) in THF (270 mL) and water (90 mL) cooled to 0 °C were added 30% hydrogen peroxide (16.36 g, 144.26 mmol, 14.87 mL, purity 30%) and lithium hydroxide (1.03 g, 42.90 mmol). The solution was stirred at 0 °C for 0.75 h, and 400 ml of a saturated solution of sodium sulfate was added. The mixture was stirred for an additional 15 min, then acidified to pH = 5 (pH paper) with 1 M HCl and extracted with ethyl acetate (200 mL × 3). The combined organics were dried over anhydrous sodium sulfate, filtered, and evaporated under reduced pressure. The residue was purified by silica flash chromatography eluting with a concentration gradient of 0 to 40% ethyl acetate in petroleum ether containing 1% acetic acid. The pure fractions were lyophilized to give (2S)-3-(tert-butoxycarbonylamino)-2-(3-methoxyphenyl)propanoic acid I-25 (8.7 g, 89% yield) as a white solid. 1 H NMR: (500 MHz, DMSO-d 6 ) δ 12.33 (s, 1H), 7.24 (t, J = 7.9 Hz, 1H), 3.79 - 3.63 (m, 4H), 3.74 - 3.71 (m, 4H) 3.42 (ddd, J = 13.3, 7.8, 5.4 Hz, 1H), 3.21 (dt, J = 13.3, 6.7 Hz, 1H), 1.33 (s, 9H). SFCt R = 4.638 min, ee value = 98.7%.

[0252] Synthesis of Intermediate I-26

[0253]

Chem.

[0254]

[0249] Step 1: To a stirred mixture of 4-bromo-benzenacetonitrile I-26a (10 g, 51.01 mmol, 1 equiv) in THF (200 mL) was added sodium hydride (2.45 g, 102.02 mmol, 2 equiv) at 0 °C. The resulting mixture was stirred at 0 °C for 2 h under an air atmosphere. Dimethyl carbonate (18.38 g, 204.03 mmol, 4 equiv) was then added to the above mixture. After the reaction, the resulting mixture was quenched with ice water (200 mL). The resulting mixture was extracted with ethyl acetate (3 × 100 mL). The combined organic layers were dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica column chromatography eluting with PE / EA (10:1, v / v) to give methyl 2-(4-bromophenyl)-2-cyanoacetate I-26b (6.1 g, 47.07%) as a yellow oil. 1 H NMR (DMSO-d 6 , 400 MHz) δ 3.74 (3H, s), 5.74 (1H, s), 7.41 (2H, m), 7.68 (2H, m)

[0250] Step 2: To a stirred mixture of nitrile I-26b (5.0 g, 19.68 mmol, 1 equiv) in methanol (35 mL) were added CoCl 2 ·6H 2 O (1102.40 mg, 39.36 mmol, 2 equiv), (Boc) 2 O (8589.67 mg, 39.36 mmol, 2 equiv), and at 0 °C, NaBH 4 (4466.65 mg, 118.07 mmol, 6 equiv) was added. The resulting mixture was stirred at 25 °C for 4 h under an air atmosphere. After the reaction, the resulting mixture was quenched by the addition of ice water (100 mL). The resulting mixture was extracted with ethyl acetate (3 × 70 mL). The combined organic layers were dried over anhydrous Na 2 SO 4It was dehydrated. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica column chromatography eluting with PE / EA (5:1, volume / volume) to obtain methyl 2-(4-bromophenyl)-3-[(tert-butoxycarbonyl)amino]propanoate I-26c (4.0 g, 56.74%) as a yellow oil. 1 H NMR (DMSO-d 6 , 400 MHz) δ 1.34 (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).

[0251] Step 3: To a stirred mixture of ester I-26c (4.0 g, 11.17 mmol, 1 equivalent) in methanol (16 mL) was added LiOH·H 2 O (1874.09 mg, 44.66 mmol, 4 equivalents) in water (4 mL). The resulting mixture was stirred at 60 °C for 8 h under a nitrogen atmosphere. After cooling to 25 °C, the resulting mixture was concentrated under reduced pressure. The reaction mixture was diluted with water (20 mL) and the pH was adjusted to 2 with HCl (1 M, 30 mL). The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, phase A, water. Phase B ACN, 0% to 50% concentration gradient over 40 min; detector, UV254 nm and hold at 30% for 20 min. After evaporation of the solvent, 2-(4-bromophenyl)-3-[(tert-butoxycarbonyl)amino]propanoic acid I-26 (3.2 g, 83.26%) was obtained as a yellow solid. LCMS: m / z (ESI), [M- t Bu] + =289.95.

[0255] Synthesis of Intermediate I-27

[0256]

Chemical formula

[0257]

[0252] Step 1: 1-Bromo-4-(2-methoxyethoxy)benzene I-27a (1 g, 4.35 mmol, 1 equiv), methyl cyanoacetate (198.18 mg, 2.00 mmol, 0.46 equiv), and Pd(OAc) 2 (97.61 mg, 0.44 mmol, 0.1 equiv), t-BuOK (975.77 mg, 8.70 mmol, 2 equiv) in dioxane (15 mL) were stirred, and Dppf (480.33 mg, 0.87 mmol, 0.2 equiv) was added at room temperature under an air atmosphere. The resulting mixture was stirred at 70 °C overnight under a nitrogen atmosphere. The resulting mixture was extracted with ethyl acetate (3 × 20 mL), and anhydrous Na 2 SO 4 was used for dehydration. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC using 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. 1 1H NMR (CDCl 3 , 400 MHz) δ 3.48 (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).

[0253] Step 2: Methyl 2-cyano-2-[4-(2-methoxyethoxy)phenyl]acetate (300 mg, 1.20 mmol, 1 equiv) and CoCl 2 ·6H 2 O (572.69 mg, 2.41 mmol, 2 equiv), and (Boc) 2 O (525.34 mg, 2.41 mmol, 2.00 equiv) in methanol (10 mL) were stirred, and NaBH 4 (364.24 mg, 9.63 mmol, 8 equiv) was added portionwise at 0 °C under an air atmosphere. The mixture in methanol was stirred at 0 °C overnight under an air atmosphere. The reaction was quenched with water (10 mL) at room temperature. The resulting mixture was extracted with ethyl acetate (3 × 15 mL), and anhydrous Na 2 SO 4 was used for dehydration. After filtration, the filtrate was concentrated under reduced pressure. The residue was in CH2 Cl 2 Purified by preparative TLC using CHCl₃ / methanol (15:1, v / v) to obtain methyl 3-[(tert-butoxycarbonyl)amino]-2-[4-(2-methoxyethoxy)phenyl]propanoate I-27b (100 mg, 23.51%) as a white solid. LCMS: m / z (ESI), [M+H-boc] + =254.

[0258]

[0254] Step 3: To a stirred mixture of ester I-27b (200 mg, 0.57 mmol, 1 equiv) in THF (40 mL) and H₂O (10 mL), LiOH (40.66 mg, 1.70 mmol, 3 equiv) was added portionwise at 0 °C under a nitrogen atmosphere. The resulting mixture was stirred at 60 °C overnight under a nitrogen atmosphere. The mixture was cooled to room temperature. The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, MeCN in water (0.1% FA), 10% to 50% concentration gradient in 10 min; detector, UV 254 nm. This gave 3-[(tert-butoxycarbonyl)amino]-2-[4-(2-methoxyethoxy)phenyl]propanoic acid I-27 (90 mg, 46.86%) as a white solid. LCMS: m / z (ESI), [M+H-boc] 2 =240. + =240.

[0259] Synthesis of Intermediate I-28

[0260]

Chemical Structure

[0261]

[0255] Step 1: To a solution of 2-(4-methoxyphenyl)acetonitrile I-28a (6 g, 40.77 mmol, 1 equiv) in THF (100 mL) was added 60% sodium hydride in oil (1.47 g, 61.15 mmol, 1.5 equiv) at 0 °C. The mixture was stirred for 80 minutes. Dimethyl carbonate (29.38 g, 326.14 mmol, 8 equiv) was added at 0 °C, and the mixture was warmed to room temperature and stirred for 4 hours. The reaction mixture was quenched with water (100 mL) and extracted with DCM (3 × 100 mL). The residue was purified by silica column chromatography eluting with PE / EA (8:1, v / v) to afford methyl 2-cyano-2-(4-methoxyphenyl)acetate (5.96 g, 70.03%) as a yellow oil.

[0262]

[0256] Step 2: NaBH 4 (40.56 g, 107.21 mmol, 4 equiv) was added portionwise to a mixture of methyl 2-cyano-2-(4-methoxyphenyl)acetate (5.50 g, 26.80 mmol, 1 equiv), (Boc) 2 O (11.7 g, 53.60 mmol, 2 equiv) and CoCl 2 ·6H 2 O (8.29 mg, 34.84 mmol, 1.3 equiv) in methanol (10 mL), and 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 × 40 mL). The combined organic layers were washed with saturated brine (1 × 100 mL) and dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica column chromatography eluting with PE / EA (5:1, v / v) to afford methyl 3-[(tert-butoxycarbonyl)amino]-2-(4-methoxyphenyl)propanoate I-28b (3.50 mg, 38.16%) as a colorless oil. LCMS: m / z (ESI), [M+H] + = 310.15.

[0263]

[0257] Step 3: In a 50 mL round-bottom flask, at room temperature, ester I-28b (2 g, 6.47 mmol, 1 equivalent) and LiOH·H 2 O (0.54 g, 12.93 mmol, 2 equivalents) as well as H 2 O (4 mL) and methanol (8 mL) were added. The resulting mixture was stirred at room temperature for 2 hours under an air atmosphere. The residue was acidified to pH = 5 with HCl (1 M). The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, methanol in water, concentration gradient from 0% to 15% in 30 minutes; detector, UV 220 nm. The resulting liquid was dried by lyophilization. Thereby, 3-[(tert-butoxycarbonyl)amino]-2-(4-methoxyphenyl)propanoic acid I-28 (1.38 g, 70.83%) was obtained as a white solid. LCMS: m / z (ESI), [M + H - t Bu] + = 240.25.

[0264] Synthesis of Intermediate I-29

[0265]

Chemical Structure

[0266]

[0258] Step 1: To a stirred solution of 2-(6-methylpyridin-2-yl)acetonitrile I-29a (500 mg, 3.78 mmol, 1 equivalent) in THF (20 mL), sodium hydride (181.57 mg, 7.57 mmol, 2 equivalents) was added portionwise at 0 °C under an air atmosphere. The resulting mixture was stirred at room temperature for 1 hour under an air atmosphere. Dimethyl carbonate (436.34 mg, 4.54 mmol, 1.2 equivalents) was added to the above mixture at room temperature. The resulting mixture was stirred for an additional 2 hours at room temperature. The reaction was quenched at room temperature by the addition of water / ice (20 mL). The resulting mixture was extracted with CH 2 Cl 2 (3 × 20 mL). The combined organic layers were dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure. The residue was taken up in CH 2 Cl2 Purified by preparative TLC using dichloromethane / methanol (20:1, volume / volume) to obtain 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 (CDCl 3 , 400 MHz) δ 2.49 (3H, s), 3.82 (3H, s), 6.49 (1H, d), 7.18 (1H, d), 7.54 (1H, dd).

[0259] Step 2: To a stirred solution of methyl 2-cyano-2-(6-methylpyridin-2-yl)acetate (1.8 g, 9.46 mmol, 1 equiv) in methanol (20 mL) was added dropwise CoCl 2 ·6H 2 O (6.75 g, 28.39 mmol, 3 equiv) and (Boc) 2 O (6.20 g, 28.39 mmol, 3 equiv) at room temperature under an air atmosphere. To the above mixture was added dropwise NaBH 4 (2.86 g, 75.71 mmol, 8 equiv) at 0 °C. The resulting mixture was stirred additionally overnight at room temperature. The reaction was quenched at room temperature by the addition of water (20 mL). The resulting mixture was extracted with CH 2 Cl 2 (3 × 20 mL). The resulting mixture was concentrated under reduced pressure. The residue was purified by preparative TLC using CH 2 Cl 2 / methanol (50:1, volume / volume) to obtain methyl 3-[(tert-butoxycarbonyl)amino]-2-(6-methylpyridin-2-yl)propanoate I-29b (1 g, 35.90%) as a yellow oil. 1 H NMR (DMSO-d 6 , 400 MHz) δ 1.34 (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).

[0260] Step 3: To a stirred solution of ester I-29b (1 g, 3.40 mmol, 1 equiv) in methanol (20 mL) and H 2 O (10 mL) was added LiOH (0.16 g, 6.69 mmol, 1.97 equiv) at room temperature. The resulting mixture was stirred at 60 °C for 2 h under an air atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, MeCN in water, 10% to 20% concentration gradient over 10 min; detector, UV 254 nm to give 3-[(tert-butoxycarbonyl)amino]-2-(6-methylpyridin-2-yl)propanoic acid I-29 (865 mg, 90.83%) as a yellow solid. 1 H NMR (DMSO-d 6 , 400 MHz) δ 1.34 (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). Synthesis of Intermediate I-30

[0267]

Chemical formula

[0268]

[0261] Step 1: To a stirred mixture of ethyl cyanoacetate (3 g, 26.52 mmol, 1 equiv) and bromocyclopentane I-30a (4.74 g, 31.83 mmol, 1.2 equiv) in DMSO (50 mL) was added K 2 CO 3 (11.00 g, 79.56 mmol, 3 equiv) under an air atmosphere at room temperature. The resulting mixture was stirred at 60 °C for 2 h under an air atmosphere. After cooling to room temperature, the reaction was quenched by the addition of water (20 mL) at room temperature. The resulting mixture was extracted with ethyl acetate (3 × 20 mL). The combined organic layers were washed with brine (3 × 20 mL) and anhydrous Na 2 SO 4It was dehydrated. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, MeCN in water, concentration gradient from 40% to 50% in 10 minutes; detector, UV254 nm to obtain ethyl 2-cyano-2-cyclopentylacetate (1.1 g, 22.89%) as a yellow oil. 1 H NMR (DMSO-d 6 , 400 MHz) δ 1.23 (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).

[0262] Step 2: To a stirred mixture of ethyl 2-cyano-2-cyclopentylacetate (900 mg, 4.97 mmol, 1 equivalent) and CoCl 2 ·6H 2 O (3.54 g, 14.90 mmol, 3 equivalents) in methanol (20 mL), (Boc) 2 O (3.25 g, 14.90 mmol, 3 equivalents) was added dropwise at room temperature under an air atmosphere. NaBH 4 (1.50 g, 39.73 mmol, 8 equivalents) was added portionwise to the above mixture over 3 minutes at 0 °C. The resulting mixture was stirred overnight at room temperature under an air atmosphere. The reaction was quenched at 0 °C by the addition of water / ice (20 mL), and the mixture was concentrated under reduced pressure. The resulting mixture was extracted with ethyl acetate (3 × 20 mL). The combined organic layers were washed with brine (3 × 20 mL) and dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica column chromatography eluting with PE / EA (1:1, volume / volume) to obtain ethyl 3-[(tert-butoxycarbonyl)amino]-2-cyclopentylpropanoate I-30b (900 mg, 63.51%) as a yellow oil. 1 H NMR (DMSO-d 6, 400 MHz) δ 1.18 (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).

[0263] Step 3: To a stirred solution of ester I-30b (1.1 g, 3.85 mmol, 1 equiv) in methanol (10 mL) and H 2 O (5 mL) was added LiOH (184.63 mg, 7.71 mmol, 2 equiv) at room temperature under an air atmosphere. The resulting mixture was stirred at 60 °C for 2 h under an air atmosphere. After cooling to room temperature, the resulting mixture was concentrated under reduced pressure. The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, MeCN in water, 20% to 30% concentration gradient over 10 min; detector, UV 254 nm to give 3-[(tert-butoxycarbonyl)amino]-2-cyclopentylpropanoic acid I-30 (878 mg, 88.52%) as a white solid. 1 H NMR (DMSO-d 6 , 400 MHz) δ 1.36 (9H, s), 1.57 (8H, d), 1.83 (1H, s), 3.00 (2H, s), 6.52 (1H, s). Synthesis of Intermediate I-31

[0269]

Chemical formula

[0270]

[0264] Step 1: In a 50 mL round-bottom flask, at room temperature, methyl 3-[(tert-butoxycarbonyl)amino]-2-(3-cyanophenyl)propanoate I-3d (300 mg, 0.99 mmol, 1 equiv) in methanol (10 mL), NH 3 aqueous solution (2.5 mL, 64.20 mmol, 65.13 equiv) and Raney Ni (10 mg, 0.12 mmol, 0.12 equiv) were added. The resulting mixture was H 2It was stirred overnight at room temperature under an atmosphere. The resulting mixture was filtered, and the filtered cake was washed with methanol (2 × 20 mL). The filtrate was concentrated under reduced pressure to obtain methyl 2-[3-(aminomethyl)phenyl]-3-[(tert-butoxycarbonyl)amino]propanoate I-31b (260 mg, 77.94%) as a green oil. LCMS: m / z (ESI), [M+H] + = 309.15.

[0271]

[0265] Step 2: To a solution of amine I-31b (240 mg, 0.78 mmol, 1 equiv) and paraformaldehyde (1402.09 mg, 15.56 mmol, 20 equiv) in DCM (100 mL), N 2 triethylamine (157.51 mg, 1.56 mmol, 2 equiv) and MgSO 4 (93.67 mg, 0.78 mmol, 1 equiv) were added under an atmosphere at 25 °C. The mixture was stirred at 25 °C for 30 minutes. Then, NaBH(OAc) 3 (659.78 mg, 3.11 mmol, 4 equiv) was added to the resulting mixture at 25 °C. The resulting mixture was stirred at 25 °C for 12 hours. The resulting mixture was filtered, and the filtered cake was washed with methanol (3 × 100 mL). The filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC using PE / EA (3:1, volume / volume) to obtain methyl 3-[(tert-butoxycarbonyl)amino]-2-{3-[(dimethylamino)methyl]phenyl}propanoate I-31c (115 mg, 37.24%) as a yellow oil. LCMS: m / z (ESI), [M+H] + = 337.20.

[0272]

[0266] Step 3: To a solution of ester I-31c (105 mg, 0.31 mmol, 1 equiv)) in THF (20 mL) and H 2 O (4 mL), N 2 LiOH·H 2O (26.19 mg, 0.62 mmol, 2 eq) was added. The mixture was stirred at 25 °C for 2 h. The resulting mixture was concentrated under reduced pressure. Thereby, 3-[(tert-butoxycarbonyl)amino]-2-{3-[(dimethylamino)methyl]phenyl}propanoic acid I-31 (100 mg, 69.15%) was obtained as a white solid. LCMS: m / z (ESI), [M+H] + = 323.20.

[0273] Synthesis of Intermediate I-32

[0274] [Chemical formula]

[0275]

[0267] Step 1: To a stirred mixture of bromide I-26b (1.0 g, 3.94 mmol, 1 eq) in 1,4-dioxane (10 mL) were added dimethylphosphine oxide (614.37 mg, 7.87 mmol, 2 eq), Pd(OAc) 2 (176.72 mg, 0.79 mmol, 0.2 eq), Xantphos (683.20 mg, 1.18 mmol, 0.3 eq) and K 3 PO 4 (2088.54 mg, 9.84 mmol, 2.5 eq). The resulting mixture was stirred at 80 °C for 2 h under a nitrogen atmosphere. After cooling to 25 °C, the resulting mixture was diluted with water (40 mL). The resulting mixture was extracted with ethyl acetate (3 × 50 mL). The combined organic layers were washed with water (3 × 30 mL) and dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure. Then it was diluted with DMF (5 mL). The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, MeCN in water, 10% to 50% concentration gradient in 10 min; detector, UV 220 nm and held at 15% for 15 min. After evaporation of the solvent, 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.

[0276]

[0268] Step 2: Methyl 2-cyano-2-[4-(dimethylphosphoryl)phenyl]acetate (660 mg, 2.63 mmol, 1 equiv), CoCl 2 ·6H 2 O (1875.16 mg, 7.881 mmol, 3 equiv), (Boc) 2 O (1146.75 mg, 5.25 mmol, 2 equiv) in MeOH (40 mL) was added portionwise with stirring to the mixture at 0 °C under a nitrogen atmosphere. NaBH 4 (795.08 mg, 21.02 mmol, 8 equiv) was added. The resulting mixture was stirred overnight. After the reaction, the reaction mixture was quenched by the addition of ice water (40 mL). The resulting mixture was extracted with DCM (3 × 40 mL). The combined organic layers were dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated 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-[4-(dimethylphosphoryl)phenyl]propanoate I-32b (330 mg, 35.35%) as a yellow oil. LCMS: m / z (ESI), [M + H - t Bu] + = 300.10.

[0277]

[0269] Step 3: To the stirred mixture of ester I-32b (310 mg, 0.87 mmol, 1 equiv) in methanol (4 mL) was added LiOH·H 2O (146.41 mg, 3.49 mmol, 4 eq) was added dropwise. 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 reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, phase B MeCN, phase A water, 10% to 50% concentration gradient in 10 min; detector, UV254 nm and held at 16.5% for 20 min. After lyophilization, 3-[(tert-butoxycarbonyl)amino]-2-[4-(dimethylphosphoryl)phenyl]propanoic acid I-32 (200 mg, 67.17%) was obtained as a white solid. 1 H NMR (DMSO-d 6 , 400 MHz) δ 1.33 (9H, s), 1.60 (6H, d), 3.27 (2H, m), 7.37 (2H, dd), 7.60 (2H, m). Synthesis of Intermediate I-33

[0278]

Chemical Structure

[0279]

[0270] Step 1: To a stirred solution of 2-(pyridin-2-yl)acetonitrile I-33a (2.0 g, 16.93 mmol, 1.00 eq) in THF (30 mL) was added sodium hydride (812.53 mg, 33.86 mmol, 2 eq) at 0 °C. The resulting mixture was stirred at 0 °C and then stirred for 30 min under a nitrogen atmosphere. Dimethyl carbonate (3049.90 mg, 33.86 mmol, 2 eq) was added dropwise to the above mixture at 0 °C. The resulting mixture was stirred for an additional 3 h at 25 °C. The reaction was quenched by the addition of water (80 mL) at 0 °C. The resulting mixture was extracted with ethyl acetate (3 × 60 mL). The combined organic layers were washed with water (3 × 40 mL) and anhydrous Na 2 SO 4It was dehydrated. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica column chromatography eluted with DCM / methanol (6%), and methyl 2-cyano-2-(pyridin-2-yl)acetate (2.3 g, 77.12%) was obtained as a yellow solid. LCMS: m / z (ESI), [M+H] + = 176.95. 1 H NMR (DMSO-d 6 , 400 MHz) δ 3.69 (3H, s), 6.88 (1H, t), 7.21 (1H, d), 7.84 (1H, ddd), 8.09 (1H, t).

[0271] Step 2: To a stirred mixture of methyl 2-cyano-2-(pyridin-2-yl)acetate (2.25 g, 12.77 mmol, 1 equiv), CoCl 2 ·6H 2 O (6077.14 mg, 25.54 mmol, 2 equiv) and (Boc) 2 O (5574.69 mg, 25.54 mmol, 2 equiv) in methanol (50 mL) was added NaBH 4 (2898.85 mg, 76.63 mmol, 6 equiv) at 0 °C. The resulting mixture was stirred at 25 °C for 3 h under an air atmosphere. The reaction was quenched at 0 °C by the addition of water (80 mL). After filtration, the resulting mixture was extracted with DCM (60 mL × 3). The combined organic layers were washed with water (3 × 50 mL) and dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC using PE / EA (3 / 1, v / v) to give methyl 3-[(tert-butoxycarbonyl)amino]-2-(pyridin-2-yl)propanoate I-33b (600 mg, 16.76%) as a yellow solid. LCMS: m / z (ESI), [M+H] + = 281.05. 1 H NMR (DMSO-d 6, 400 MHz) δ 1.34 (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).

[0272] Step 3: To a stirred mixture of ester I-33b (575 mg, 2.05 mmol, 1 equiv) in methanol (4.0 mL) was added LiOH (196.50 mg, 8.20 mmol, 4 equiv) in water (1.0 mL) at 25 °C. The resulting mixture was stirred at 70 °C for 3 h under an air atmosphere. After cooling to room temperature, the resulting mixture was concentrated under vacuum 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). The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, phase B MeCN, phase A water, 10% to 50% concentration gradient in 10 min; detector, UV254 nm and held at 10.8% for 20 min. After lyophilization, 3-[(tert-butoxycarbonyl)amino]-2-(pyridin-2-yl)propanoic 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-d 6 , 400 MHz) δ 1.33 (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). Synthesis of Intermediate I-34

[0280]

Chem.

[0281]

[0273] Step 1: A solution of (4-bromo-2-methoxyphenyl)methanol I-34a (2 g, 9.21 mmol, 1 equiv) in CH 2 Cl 2 was treated with triethylamine (2.80 g, 27.64 mmol, 3 equiv) and DMAP (11.26 mg, 0.092 mmol, 0.01 equiv) at 0 °C for 5 min under an air atmosphere, followed by the dropwise addition of TBSCl (1.67 g, 11.06 mmol, 1.2 equiv) at 0 °C. The resulting mixture was stirred at room temperature for 1 h under an air atmosphere. The resulting mixture was diluted with water (100 mL). The aqueous layer was extracted with CH 2 Cl 2 (3 × 100 mL). The combined organic layers were concentrated under reduced pressure. The residue was purified by preparative TLC using PE / EA (40:1, v / v) to give [(4-bromo-2-methoxyphenyl)methoxy](tert-butyl)dimethylsilane (2.56 g, 83.86%) as a yellow oil. 1 H NMR (CD 3 OD, 400 MHz) δ 0.12 (6H, s), 0.97 (9H, s), 3.84 (3H, s), 4.70 (2H, d), 7.06 - 7.14 (2H, m), 7.31 (1H, dt).

[0274] Step 2: In a 40 mL vial, [(4-bromo-2-methoxyphenyl)methoxy](tert-butyl)dimethylsilane (1 g, 3.02 mmol, 1 equiv), methyl cyanoacetate (358.88 mg, 3.62 mmol, 1.20 equiv), Pd(OAc) 2 (67.76 mg, 0.30 mmol, 0.1 equiv), dppf (333.43 mg, 0.60 mmol, 0.2 equiv), and t-BuOK (1.02 g, 9.05 mmol, 3 equiv) in 1,4-dioxane (8 mL) were added at room temperature. The resulting mixture was stirred at 90 °C overnight under a nitrogen atmosphere. The mixture was acidified to pH 5 with saturated NH 4 Cl (aqueous solution). The resulting mixture was stirred at room temperature for 30 min. The aqueous layer was extracted with ethyl acetate (3 × 100 mL). The combined organic layers were dried over anhydrous Na 2 SO 4It was dehydrated, filtered, and concentrated under reduced pressure. The residue was purified by preparative TLC using PE / EA (4:1, volume / volume) to obtain 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-d 6 , 400 MHz) δ 0.09 (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).

[0275] Step 3: In a 40 mL vial, at 0 °C, nitrile I-34b (550 mg, 1.57 mmol, 1 equivalent) in methanol (5 mL), CoCl 2 ·6H 2 O (973.46 mg, 4.09 mmol, 2.6 equivalents) and Boc 2 O (1.37 g, 6.30 mmol, 4 equivalents) were added. NaBH 4 (476.25 mg, 12.59 mmol, 8 equivalents) was added to the above mixture at 0 °C. The resulting mixture was stirred at room temperature for 2 hours. The reaction was quenched with water at room temperature. The resulting mixture was filtered, and the filter cake was washed with DCM (3 × 30 mL). The filtrate was extracted with DCM (3 × 30 mL). The combined organic layers were dehydrated with anhydrous Na 2 SO 4 and filtered, then concentrated under reduced pressure. The residue was purified by preparative TLC using CH 2 Cl 2 / methanol (40:1, volume / volume) to obtain methyl 3-[(tert-butoxycarbonyl)amino]-2-(4-{[(tert-butyldimethylsilyl)oxy]methyl}-3-methoxyphenyl)propanoate (410 mg, 57.43%) as a yellow oil. LCMS: m / z (ESI), [M+Na] + = 476.15. 1 H NMR (DMSO-d 6, 400 MHz) δ 0.08 (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).

[0276] Step 4: In a 50 mL round-bottom flask, at room temperature, methyl 3-[(tert-butoxycarbonyl)amino]-2-(4-{[(tert-butyldimethylsilyl)oxy]methyl}-3-methoxyphenyl)propanoate (390 mg, 0.86 mmol, 1 equivalent), THF (2 mL, 24.69 mmol, 28.71 equivalents) and LiOH·H 2 O (72.15 mg, 1.72 mmol, 2.0 equivalents) were added. The resulting mixture was stirred at room temperature for 1.5 hours. The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, methanol in water with a concentration gradient from 0% to 100% in 30 minutes; detector, UV 220 nm. The resulting mixture was concentrated under vacuum to obtain 3-[(tert-butoxycarbonyl)amino]-2-(4-{[(tert-butyldimethylsilyl)oxy]methyl}-3-methoxyphenyl)propanoic acid I-34 (160 mg, 42.33%) as a gray solid. LCMS: m / z (ESI), [M+Na] + = 462.15.

[0282] Synthesis of Intermediate I-35

[0283]

Chemical Structure

[0284]

[0277] Step 1: In a 100 mL round-bottom flask, 3-bromo-5-methoxypyridine I-35a (4 g, 21.27 mmol, 1 eq) and methyl cyanoacetate (2.53 g, 25.53 mmol, 1.2 eq) in dioxane (40 mL, 590.14 mmol, 27.74 eq) at room temperature, Pd(OAc) 2 (477.62 mg, 2.13 mmol, 0.1 eq), dppf (2.35 g, 4.26 mmol, 0.2 eq), t-BuOK (7.16 g, 63.82 mmol, 3 eq) were added. The resulting mixture was stirred at 70 °C for 5 h under a nitrogen atmosphere. The mixture was acidified to pH 6 with saturated NH 4 Cl (aqueous solution). The resulting mixture was extracted with ethyl acetate (3 × 60 mL). The combined organic layers were washed with DCM (3 × 3 mL) and dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica column chromatography eluting with CH 2 Cl 2 / methanol (50:1, v / v) 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.

[0285]

[0278] Step 2: To a stirred mixture of methyl 2-cyano-2-(5-methoxypyridin-3-yl)acetate (700 mg, 3.40 mmol, 1 eq) and CoCl 2 ·6H 2 O (1.21 g, 5.09 mmol, 1.5 eq) in methanol (15 mL) at 0 °C under a nitrogen atmosphere, NaBH 4 (385.27 mg, 10.19 mmol, 3 eq) was added portionwise. The resulting mixture was stirred at room temperature for 1 h 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 × 10 mL). The resulting mixture was extracted with DCM (3 × 100 mL). The combined organic layers were washed with DCM (2 × 3 mL) and dried over anhydrous Na 2 SO 4It was dehydrated. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC using CH 2 Cl 2 / methanol (35:1, v / v) to obtain methyl 3-[(tert-butoxycarbonyl)amino]-2-(5-methoxypyridin-3-yl)propanoate I-35b (250 mg, 23.73%) as a pale yellow solid. LCMS: m / z (ESI), [M+H] + = 311.15.

[0286]

[0279] Step 3: In a 50 mL round-bottom flask, ester I-35b (80 mg, 0.26 mmol, 1 equivalent) and LiOH (12.35 mg, 0.52 mmol, 2 equivalents) in water (3 mL) and methanol (3 mL) were added at room temperature. The resulting mixture was stirred at room temperature for 2 hours under a nitrogen atmosphere. The mixture was acidified to pH 5 with concentrated HCl. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, methanol in water, concentration gradient from 28% to 32% in 15 minutes; detector, UV 254 nm. The resulting mixture was concentrated under vacuum. Thereby, 3-[(tert-butoxycarbonyl)amino]-2-(5-methoxypyridin-3-yl)propanoic acid I-35 (40 mg, 52.37%) was obtained as an off-white solid. LCMS: m / z (ESI), [M+H] + = 297.05.

[0287] Synthesis of Intermediate I-36

[0288]

Chemical formula

[0289]

[0280] Step 1: In a 250 mL round-bottom flask, 4-bromo-1-methylpyridin-2-one I-36a (3 g, 15.96 mmol, 1 equivalent) and methyl cyanoacetate (1.90 g, 19.15 mmol, 1.2 equivalents) and Pd(OAc) 2(358.21 mg, 1.60 mmol, 0.1 eq) and Dppf (1.76 g, 3.19 mmol, 0.2 eq) and t-BuOK (5.37 g, 47.87 mmol, 3 eq) and 1,4-dioxane (100 mL) were added. The resulting mixture was stirred at 70 °C for 1 h 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 ethyl acetate 3 × 100 mL. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in methanol (5 mL). The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, methanol in water, concentration gradient from 0% to 0% in 15 min; detector, UV 254 nm. The resulting mixture was concentrated under reduced pressure. Thereby, methyl 2-cyano-2-(1-methyl-2-oxopyridin-4-yl)acetate (1.15 g, 34.95%) was obtained as a brown solid. LCMS: m / z (ESI), [M+H] + = 207.05. 1 H NMR (CD 3 OD, 400 MHz) δ 3.37 (1H, s), 3.45 (3H, s), 3.63 (3H, s), 6.75 (1H, s), 6.91 (1H, s), 7.23 (1H, d).

[0281] Step 2: A solution of methyl 2-cyano-2-(1-methyl-2-oxopyridin-4-yl)acetate (1.1 g, 5.34 mmol, 1 eq) in methanol (15 mL) was treated with Boc 2 O (4.66 g, 21.34 mmol, 4 eq) and CoCl 2 ·6H 2 O (3.30 g, 13.87 mmol, 2.6 eq) at 0 °C for 5 min under an air atmosphere, followed by the addition of NaBH 4 (1.61 g, 42.68 mmol, 8 eq) portionwise at 0 °C. The resulting mixture was stirred at 70 °C for 15 h under an air atmosphere. The reaction was quenched by the addition of water (80 mL) at room temperature. The resulting mixture was filtered, and the filter cake was washed with DCM (3 × 10 mL). The aqueous layer was CH 2 Cl 2It was extracted with (3 × 80 mL). The resulting mixture was concentrated under reduced pressure. The residue was purified by preparative TLC using CH 2 Cl 2 / methanol (18:1, volume / volume) to obtain methyl 3-[(tert-butoxycarbonyl)amino]-2-(1-methyl-2-oxopyridin-4-yl)propanoate I-36b (445 mg, 26.88%) as a white oil. LCMS: m / z (ESI), [M+H] + = 311.15.

[0290]

[0282] Step 3: In a 100 mL round-bottom flask, at room temperature, ester I-36b (445 mg, 1.43 mmol, 1 equivalent) and LiOH·H 2 O (120.33 mg, 2.87 mmol, 2 equivalents) as well as THF (3 mL) and H 2 O (3 mL) were added. The resulting mixture was stirred at room temperature for 2 hours under an air atmosphere. The mixture was acidified to pH 5 with HCl (1 M). The resulting mixture was concentrated under reduced pressure. The residue was dissolved in methanol (5 mL). The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, methanol in water, concentration gradient from 0% to 10% in 30 minutes; detector, UV220 nm. The resulting mixture was concentrated under reduced pressure. The resulting liquid was dried by lyophilization. Thereby, 3-[(tert-butoxycarbonyl)amino]-2-(1-methyl-2-oxopyridin-4-yl)propanoic acid I-36 (350 mg, 82.37%) was obtained as an off-white solid. LCMS: m / z (ESI), [M+H] + = 297.15. 1 H NMR (CD 3 OD, 400 MHz) δ 1.43 (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). Synthesis of Intermediate I-37

[0291]

Chemical Structure

[0292]

[0283] Step 1: To a stirred solution of 4-bromo-benzenacetonitrile I-37a (10 g, 51.01 mmol, 1 equiv) in THF (120 mL) at 0 °C was added sodium hydride (4 g, 166.68 mmol, 3.27 equiv). The resulting mixture was stirred at 0 °C for 1.5 h under an air atmosphere. Then dimethyl carbonate (18.38 g, 204.03 mmol, 4 equiv) was added to the above mixture at 0 °C. After the reaction, the resulting mixture was diluted with water (20 mL). The resulting mixture was extracted with ethyl acetate (3 × 30 mL). The combined organic layers were dried over anhydrous Na 2 SO 4 4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC using PE / EA (2:1, v / v) to give methyl 2-(4-bromophenyl)-2-cyanoacetate (8.923 g, 68.85%) as a yellow oil. 1 1H NMR (CDCl 3 3, 400 MHz) δ 3.84 (3H, s), 4.72 (1H, s), 7.35 - 7.39 (2H, m), 7.56 - 7.62 (2H, m).

[0284] Step 2: In a 250 mL round-bottom flask were added methyl 2-(4-bromophenyl)-2-cyanoacetate (2 g, 7.87 mmol, 1 equiv), Boc 2 2O (3.44 g, 15.74 mmol, 2 equiv) and CoCl 2 2·6H 2 2O (3.75 g, 15.74 mmol, 2 equiv) in methanol (70 mL) at room temperature. NaBH 4 4 (1.79 g, 47.23 mmol, 6 equiv) was added to the above mixture at 0 °C. The resulting mixture was stirred at 0 °C for 2 h. The resulting mixture was filtered and the filtered cake was washed with DCM (4 × 50 mL). The filtrate was concentrated in CH 2 2Cl 2It was extracted with (3 × 100 mL). The combined organic layers were concentrated under reduced pressure. The residue was purified by silica column chromatography eluted with PE / EA (1:1, volume / volume) to obtain methyl 2-(4-bromophenyl)-3-[(tert-butoxycarbonyl)amino]propanoate 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-d 6 , 400 MHz) δ 1.33 (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).

[0285] Step 3: In a 40 mL vial, at room temperature, ester I-37b (1.9 g, 5.30 mmol, 1 equivalent) in DMF (10 mL), Zn(CN) 2 (0.62 g, 5.30 mmol, 1.0 equivalent) and Pd(PPh 3 ) 4 (0.61 g, 0.53 mmol, 0.1 equivalent) were added. The resulting mixture was stirred at 80 °C for 3 hours under a nitrogen atmosphere. The residue was washed with saturated NaHCO 3 (1 × 40 mL). The resulting mixture was extracted with ethyl acetate (3 × 200 mL). The combined organic layers were concentrated under reduced pressure. The residue was purified by preparative TLC using PE / EA (1:1, volume / volume) to obtain methyl 3-[(tert-butoxycarbonyl)amino]-2-(4-cyanophenyl)propanoate (1.0 g, 61.95%) as a yellow solid. LCMS: m / z (ESI), [M + H - tBu] + = 248.95. 1 H NMR (DMSO-d 6 , 400 MHz) δ 1.31 (9H, s), 3.51 (1H, ddd), 3.62 (3H, s), 4.01 (2H, dt), 7.44 - 7.50 (2H, m), 7.82 (2H, d).

[0286] Step 4: In a 20 mL vial, at room temperature, methyl 3-[(tert-butoxycarbonyl)amino]-2-(4-cyanophenyl)propanoate (900 mg, 2.96 mmol, 1 equiv), methanol (3.75 mL) and LiOH·H 2 O (248.16 mg, 5.91 mmol, 2 equiv) were added. The resulting mixture was stirred at 70 °C for 1.5 h. The mixture was acidified to pH 5 with HCl (aqueous solution). The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, methanol in water, 0% to 100% concentration gradient in 30 min; detector, UV 254 nm. The resulting mixture was concentrated under vacuum to give 3-[(tert-butoxycarbonyl)amino]-2-(4-carbamoylphenyl)propanoic acid I-37 (800 mg, 87.74%) as a white solid. LCMS: m / z (ESI), [M+H- t Bu] + = 253.00.

[0293] Synthesis of Intermediate I-38

[0294]

Chemical Structure

[0295]

[0287] Step 1: To a solution of methyl 2-(4-bromophenyl)-2-[(tert-butoxycarbonyl)amino]acetate I-38a (500 mg, 1.45 mmol, 1 equiv) in dioxane (30 mL), (Bu) 3 SnCH 2 OH (700 mg, 2.18 mmol, 1.5 equiv) and Pd(PPh 3 ) 4 (84 mg, 0.073 mmol, 0.05 equiv) were added under an air atmosphere at 80 °C for 16 h. The mixture was cooled to room temperature. The resulting mixture was quenched with water and extracted with ethyl acetate (3 × 30 mL). The combined organic layers were washed with saturated NaCl (3 × 30 mL) and anhydrous Na 2 SO 4It was dehydrated. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC eluting with petroleum ether / ethyl acetate (3:1, v / v) to give methyl 2-[(tert-butoxycarbonyl)amino]-2-[4-(hydroxymethyl)phenyl]acetate I-38b (325 mg) as an orange oil (76%). LCMS: m / z (ESI), [M+H] + = 296.14. 1 H NMR (DMSO-d 6 , 400 MHz) δ 1.39 (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).

[0288] Step 2: A mixture of methyl 2-[(tert-butoxycarbonyl)amino]-2-[4-(hydroxymethyl)phenyl]acetate (390 mg, 1.30 mmol, 1 equiv) and LiOH·H 2 O (109 mg, 2.60 mmol, 2 equiv) in THF (4 mL) and H 2 O (1 mL) was stirred at room temperature for 5 h under a nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was purified by preparative HPLC using the following conditions (the mobile phase consisted of a solvent mixture of 0.1% NH 4 HCO 3 in water and 0.1% NH 4 OH in acetonitrile. A constant concentration gradient from 95% aqueous / 5% organic to 5% aqueous / 95% organic mobile phase was used over 25 min. The flow rate was constant at 40 mL / min.) to give [(tert-butoxycarbonyl)amino][4-(hydroxymethyl)phenyl]acetic acid I-38 (84%) 312 mg as a yellow solid. LCMS: m / z (ESI), [M+H] + = 282.13.

[0296] Synthesis of Intermediate I-39

[0297]

Chemical Structure

[0298]

[0289] Step 1: To a mixture of methyl 2-(3-bromophenyl)acetate I-39a (3.0 g, 13.10 mmol, 1 equiv) in DMF (4.0 mL), K 2 CO 3 (2171.96 mg, 15.72 mmol, 1.20 equiv) and polyoxymethylene (601.93 mg, 13.36 mmol, 1.02 equiv) were added. The resulting mixture was stirred at 70 °C for 2 h under a nitrogen atmosphere. After cooling to room temperature, the resulting mixture was diluted with water (40 mL). The resulting mixture was extracted with ethyl acetate (3 × 40 mL). The combined organic layers were washed with water (3 × 10 mL) and dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC using 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-d 6 , 400 MHz) δ 3.76 (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).

[0290] Step 2: To a mixture of methyl 2-(3-bromophenyl)prop-2-enoate (800 mg, 3.32 mmol, 1 equiv) in DMF (5 mL), 1-methylpiperazine (1661.86 mg, 16.59 mmol, 5 equiv) was added. The resulting mixture was stirred at 25 °C for 2 h under a nitrogen atmosphere. The reaction mixture was diluted with EA (20 mL). The residue was washed with saturated NaHCO 3 (2 × 10 mL) then water (2 × 20 mL) and saturated NaCl (3 × 20 mL). The resulting organic layer was dried over Na 2 SO 4It was dried. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, MeCN in water, concentration gradient from 10% to 50% in 10 minutes; detector, UV254 nm, held at 65% for 30 minutes. After evaporating the water, methyl 2-(3-bromophenyl)-3-(4-methylpiperazin-1-yl)propanoate I-39b (740 mg, 65.35%) was obtained as a yellow oil. LCMS: m / z (ESI), [M+H] + = 341.00, 343.00.

[0299]

[0291] Step 3: 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), Pd(PPh 3 ) 4 (152.39 mg, 0.13 mmol, 0.1 equivalent) was added portionwise at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at 80 °C for 2.0 hours under a nitrogen atmosphere. 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 × 30 mL). The combined organic layers were washed with water (3 × 10 mL) and dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, MeCN in water, concentration gradient from 10% to 50% in 10 minutes; detector, UV254 nm, held at 28% for 30 minutes. After evaporating the solvent, methyl 2-(3-cyanophenyl)-3-(4-methylpiperazin-1-yl)propanoate (400 mg, 52.50%) was obtained as a yellow oil. LCMS: m / z (ESI), [M+H] + = 288.05.

[0300]

[0292] Step 4: Methyl 2-(3-cyanophenyl)-3-(4-methylpiperazin-1-yl)propanoate (400 mg, 1.39 mmol, 1 equivalent) in THF (8.0 mL) and LiOH·H in water (2.0 mL)2 To a stirred mixture of O (175.22 mg, 4.18 mmol, 3.0 eq), it was added dropwise at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at 25 °C for 4.0 h under a nitrogen atmosphere. After the reaction, the resulting mixture was concentrated under vacuum. The resulting mixture was diluted with methanol (4.0 mL). The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, MeCN in water, 0% to 50% concentration gradient in 10 min; detector, UV 254 nm and held at 9.8% for 20 min to obtain 2-(3-cyanophenyl)-3-(4-methylpiperazin-1-yl)propanoic acid I-39 (240 mg, 63.08%) as a white solid. LCMS: m / z (ESI), [M+H] + = 274.10.

[0301] Synthesis of Intermediate I-40

[0302]

Chemical Structure

[0303]

[0293] Step 1: In a 100 mL three-necked round-bottom flask, 3-bromo-5-methoxypyridine I-40a (8 g, 42.55 mmol, 1 eq) and THF (40 mL) were added at room temperature. Isopropylmagnesium bromide, 1 M solution in THF (17.61 mL, 51.06 mmol, 1.2 eq) was added to the mixture at -10 °C under a nitrogen atmosphere. The mixture was stirred at -10 °C for 30 min under a nitrogen atmosphere. Ethyl 2-oxobutanoate (6.64 g, 51.06 mmol, 1.2 eq) was added to the above mixture at -10 °C. The resulting mixture was stirred at -8 °C for 3 h under a nitrogen atmosphere. The reaction was quenched by the addition of saturated NH 4 Cl (aqueous solution) (50 mL) at 0 °C. The resulting mixture was extracted with ethyl acetate (3 × 30 mL). The combined organic layers were dried over anhydrous Na 2 SO 4It was dehydrated, filtered, and evaporated to obtain a crude oily substance. The residue was purified by silica column chromatography eluted with PE / EA (1:1, volume / volume) to obtain ethyl 2-(5-methoxypyridin-3-yl)-2-oxoacetate I-40b (4.6 g, 45.48%) as a yellow oily substance. LCMS: m / z (ESI), [M+H] + = 210.05. 1 H NMR (CD 3 OD, 400 MHz) δ 1.23 (3H, t), 3.91 (3H, s), 4.15 - 4.26 (2H, m), 7.61 (1H, dd), 8.23 (1H, d), 8.37 (1H, d).

[0294] Step 2: In a 50 mL round-bottom flask, at 0 °C, ester I-40b (2 g, 9.56 mmol, 1 equivalent), hydroxylamine hydrochloride (0.86 g, 12.43 mmol, 1.3 equivalents), pyridine (0.83 g, 10.52 mmol, 1.1 equivalents), and methanol (10 mL) were added. The resulting mixture was stirred at room temperature for 6 hours. The resulting mixture was diluted with CH 2 Cl 2 (40 mL). The resulting mixture was washed with HCl (0.5 M) 1×40 mL, water 1×40 mL, and saturated NaCl (aqueous solution) 1×40 mL. The organic layer was concentrated under vacuum. The solid precipitated in the aqueous phase was collected by filtration and washed with water (3×10 mL). Thereby, ethyl (2E)-2-(N-hydroxyimino)-2-(5-methoxypyridin-3-yl)acetate (1.78 g, 83.04%) was obtained as a white solid. LCMS: m / z (ESI), [M+H] + = 225.15.

[0304]

[0295] Step 3: In a 50 mL round-bottom flask, ethyl 2-(N-hydroxyimino)-2-(5-methoxypyridin-3-yl)acetate (1.7 g, 7.58 mmol, 1 eq), Pd / C (1.86 g, 17.44 mmol, 2.3 eq) and methanol (100 mL) were added at room temperature. The resulting mixture was stirred at room temperature for 6 h under a hydrogen atmosphere. The resulting mixture was filtered, and the filtered cake was washed with methanol (6 × 30 mL). The filtrate was concentrated under reduced pressure. Thereby, ethyl 2-amino-2-(5-methoxypyridin-3-yl)acetate I-40c (1.6 g, 87.33%) was obtained as a yellow oil. LCMS: m / z (ESI), [M+H] + =211.10。 1 H NMR (CD 3 OD, 400 MHz) δ 1.22 (3H, t), 4.19 (2H, m), 4.69 (1H, d), 4.88 (3H, s), 7.50 (1H, dq), 8.20 (2H, t).

[0296] Step 4: In a 50 mL round-bottom flask, amine I-40c (1.6 g, 7.61 mmol, 1 eq), triethylamine (3.85 g, 38.06 mmol, 5 eq), Boc 2 O (3.32 g, 15.22 mmol, 2 eq) and DCM (5 mL) were added at room temperature. The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was quenched with water (50 mL) and extracted with DCM (2 × 50 mL). The combined organic layers were washed with saturated brine (1 × 50 mL), and the organic layer was dehydrated with anhydrous Na 2 SO 4 4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC using CH 2 Cl 2 / methanol (25:1, v / v) to obtain ethyl 2-[(tert-butoxycarbonyl)amino]-2-(5-methoxypyridin-3-yl)acetate (1.38 g, 49.19%) as a yellow oil. LCMS: m / z (ESI), [M+H] + =311.20。 1 H NMR (CD 3OD, 400 MHz) δ 1.19 - 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).

[0297] Step 5: A solution of ethyl 2-[(tert-butoxycarbonyl)amino]-2-(5-methoxypyridin-3-yl)acetate (1.38 g, 4.45 mmol, 1 equiv) and LiOH (0.21 g, 8.89 mmol, 2 equiv) in THF (13 mL) and water (13 mL) was stirred at 60 °C for 6 h. The mixture / residue was acidified to pH 5 with concentrated HCl. The precipitated solid was collected by filtration and washed with methanol (2 × 10 mL). The filtrate was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, methanol in water, 0 to 50% concentration gradient in 30 min; detector, UV 254 nm. This gave [(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.

[0305] Synthesis of Intermediate I-41

[0306]

Chemical Structure

[0307]

[0298] Methyl 3-[(tert-butoxycarbonyl)amino]-2-(3-cyanophenyl)propanoate I-3d (700 mg, 2.30 mmol, 1 equiv) and LiOH·H 2 O (220.34 mg, 9.20 mmol, 4 equiv) in MeOH (8 mL) and H 2The mixture in O (4 mL) was stirred at 70 °C for 4.5 hours under an air atmosphere. The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica gel; mobile phase, MeCN in water, concentration gradient from 10% to 100% in 50 minutes; detector, UV 220 nm. Thereby, 3-[(tert-butoxycarbonyl)amino]-2-(3-carbamoylphenyl)propanoic acid I-41 (330 mg, 46.53%) was obtained as a yellow solid. LCMS: m / z (ESI), [M-H] + = 307.05. 1 H NMR (CD 3 OD, 400 MHz) δ 1.40 (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). Synthesis of Intermediate I-42

[0308]

Chemical Structure

[0309]

[0299] Step 1: To a stirred mixture of 2-amino-1-(pyridin-3-yl)ethanone I-42a (1.0 g, 7.35 mmol, 1 equivalent) and Et 3 N (2.23 g, 22.03 mmol, 3 equivalents) in DCM (20 mL) was added Boc 2 O (1.92 g, 8.81 mmol, 1.2 equivalents) at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at room temperature for 3 hours. After the reaction, the resulting mixture was diluted with water (50 mL) and extracted with ethyl acetate (3 × 30 mL). The combined organic layers were dehydrated with anhydrous Na 2 SO 4 4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC using PE / EA (1:1, volume / volume) to obtain tert-butyl N-[2-oxo-2-(pyridin-3-yl)ethyl]carbamate (650 mg, 37.46%) as a yellow solid. LCMS: [M+H] + = 237.05. 1 H NMR (CDCl3 , 400 MHz) δ 1.52 (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).

[0300] Step 2: To a stirred mixture of tert-butyl N-[2-oxo-2-(pyridin-3-yl)ethyl]carbamate (640 mg, 2.71 mmol, 1 equiv) and hydroxylamine hydrochloride (564.70 mg, 8.13 mmol, 3 equiv) in methanol (10 mL), pyridine (642.79 mg, 8.13 mmol, 3 equiv) was added dropwise at 0 °C under a nitrogen atmosphere. The resulting mixture was stirred at room temperature overnight. After the reaction, the resulting mixture was quenched with water (20 mL). The resulting mixture was extracted with ethyl acetate (3 × 20 mL). The combined organic layers were dried over anhydrous Na 2 SO 4 and filtered. After filtration, the filtrate was concentrated under reduced pressure to give tert-butyl N-[2-(hydroxyimino)-2-(pyridin-3-yl)ethyl]carbamate I-42b (400 mg, 58.77%) as a yellow oil. LCMS: m / z (ESI), [M+H] + = 252.00.

[0310]

[0301] Step 3: To a stirred mixture of hydroxyimine I-42b (465 mg, 1.85 mmol, 1 equiv) and formic acid (170.34 mg, 3.70 mmol, 2 equiv) in methanol (4.0 mL), Zn (362.95 mg, 5.55 mmol, 3 equiv) was added portionwise at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at room temperature for 3 h. The desired product was detected by LCMS. After the reaction, the resulting mixture was concentrated under vacuum. It was then diluted with methanol (4.0 mL). The residue was K 2 CO 3It was basified to pH 9. The precipitated solid was filtered off and washed with DCM / methanol (10:1, v / v). The filtrate was evaporated and purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, MeCN in water, 10% to 50% concentration gradient over 10 minutes; detector, UV 220 nm, held at 45% for 20 minutes. After evaporation of the solvent, tert-butyl N-[2-amino-2-(pyridin-3-yl)ethyl]carbamate I-42 (287 mg, 65.36%) was obtained as a yellow oil. LCMS: m / z (ESI), [M+H] + =238.10。 1 H NMR (DMSO-d 6 , 400 MHz) δ 1.32 (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). Synthesis of Intermediate I-43

[0311]

Chemical Structure

[0312]

[0302] Step 1: In a 250 mL round-bottom flask, [(tert-butoxycarbonyl)amino](3-methoxyphenyl)acetic acid I-43a (3 g, 10.66 mmol, 1 equiv), piperazine, 1-methyl- (1.28 g, 12.80 mmol, 1.2 equiv), TCFH (4.49 g, 16.00 mmol, 1.5 equiv), and NMI (4.38 g, 53.32 mmol, 5 equiv) in MeCN (30 mL) at room temperature were added. The resulting mixture was stirred at room temperature for 3 h under a nitrogen atmosphere. The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, methanol in water, 0% to 70% concentration gradient in 40 min; detector, UV254 nm to obtain tert-butyl N-[1-(3-methoxyphenyl)-2-(4-methylpiperazin-1-yl)-2-oxoethyl]carbamate I-43b (2.5 g, 64.50%) as a yellow oil. LCMS: m / z (ESI), [M+H] + = 364.20.

[0313]

[0303] Step 2: In a 50 mL round-bottom flask, amide I-43b (2.2 g, 6.05 mmol, 1 equiv) and TFA (4 mL, 53.85 mmol, 8.90 equiv) in DCM (4 mL) at room temperature were added. The resulting mixture was stirred at room temperature for 15 h under a nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was basified to pH 9 with NH 3 ·H 2 O. The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, methanol in water (5 mmol / L NH 4 HCO 3 ), 10% to 50% concentration gradient in 10 min; detector, UV254 nm to obtain 2-amino-2-(3-methoxyphenyl)-1-(4-methylpiperazin-1-yl)ethanone I-43c (1.5 g, 94.10%) as a yellow oil. LCMS: m / z (ESI), [M+H- t Bu] + = 264.05. 1 H NMR (CD 3OD, 400 MHz) δ 2.62 (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).

[0304] Step 3: In a 40 mL vial, amine I-43c (1.5 g, 5.70 mmol, 1 equivalent) and BH 3 ·C 4 H 8 O (20 mL, 208.98 mmol, 36.69 equivalents) were added at room temperature. The resulting mixture was stirred at 70 °C for 24 h under a nitrogen atmosphere. The mixture was cooled to room temperature. The reaction was quenched at 0 °C by the addition of methanol (20 mL). The resulting mixture was stirred for 2 h, concentrated under reduced pressure, and the residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, methanol in water, 0% to 50% concentration gradient over 40 min; detector, UV 220 nm to give 1-(3-methoxyphenyl)-2-(4-methylpiperazin-1-yl)ethanamine I-43 (500 mg, 35.20%) as a colorless oil. LCMS: m / z (ESI), [M + H - t Bu] + = 250.05.

[0314] Synthesis of Intermediate I-44

[0315]

Chemical Structure

[0316]

[0305] Step 1: A mixture of tribromomane - tetrabutylamine (6.42 g, 13.32 mmol, 1 equiv) and 1 - [4 - (hydroxymethyl)phenyl]ethanone I - 44a (2 g, 13.32 mmol, 1 equiv) in ACN (100 mL) and acetone (10 mL) was stirred at room temperature for 1 hour under a nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure. The resulting mixture was diluted with water (10 mL). The resulting mixture was extracted with MTBE (2×20 mL). The combined organic layers were washed with water (7×20 mL) and dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure to obtain 2 - bromo - 1 - [4 - (hydroxymethyl)phenyl]ethanone I - 44b (2 g, 65.56%) as a yellow oil. The resulting mixture was used directly in the next step.

[0317]

[0306] Step 2: A mixture of NH 3 (gas) (6.24 mL, 43.66 mmol, 5 equiv) and bromide I - 44b (2 g, 8.73 mmol, 1 equiv) in THF (50 mL) was stirred at room temperature for 2 hours under a nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, MeCN in water (0.1% NH 3 ·H 2 O), 0% to 50% concentration gradient in 60 minutes; detector, UV254 nm to obtain 2 - amino - 1 - [4 - (hydroxymethyl)phenyl]ethanone I - 44c (1 g, 69.34%) as a colorless oil. 1 H NMR (400 MHz, DMSO - d 6 ) δ 2.56 (2H, d), 4.62 (2H, s), 5.43 (1H, s), 7.53 (2H, d), 8.00 (2H, d), 8.21 (2H, s).

[0307] Step 3: Boc 2A mixture of O (1.32 g, 6.05 mmol, 1 equiv), triethylamine (1.84 g, 18.16 mmol, 3 equiv) and amine I-44c (1 g, 6.05 mmol, 1 equiv) in DCM (20 mL) was stirred at room temperature for 2 h under a nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was purified by preparative TLC (prep-TLC water PE / EA (1:2, v / v)) using PE / EA (1:2, v / v) to give tert-butyl N-{2-[4-(hydroxymethyl)phenyl]-2-oxoethyl}carbamate I-44d (500 mg, 31.13%) as a yellow solid. 1 H NMR (400 MHz, DMSO-d 6 ) δ 1.40 (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).

[0308] Step 4: NaBH 3 CN (278.31 mg, 4.43 mmol, 2.5 equiv), NH 4 OAc (682.77 mg, 8.86 mmol, 5 equiv) and a mixture of ketone I-44d (470 mg, 1.77 mmol, 1 equiv) in methanol (10 mL) were stirred at 80 °C for 2 h under a nitrogen atmosphere. The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, methanol in water (10 mmol / L NH 4 HCO 3 ), concentration gradient from 0% to 100% in 60 min; detector, UV254 nm to give tert-butyl N-{2-amino-2-[4-(hydroxymethyl)phenyl]ethyl}carbamate I-44 (110 mg, 23.31%) as a yellow oil. 1 H NMR (400 MHz, DMSO-d 6 ) δ 1.41 (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). Synthesis of Intermediate I-45

[0318]

Chem.

[0319]

[0309] Step 1: A solution of methyl pyruvate I-45a (5 g, 48.98 mmol, 1 equiv) and 4-methylbenzenesulfonyl hydrazide (10.03 g, 53.88 mmol, 1.1 equiv) in 1,4-dioxane (30 mL) was stirred at 70 °C for 2 h under a nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was purified by trituration with methanol (20 mL) to give methyl [N'-(4-methylbenzenesulfonyl)hydrazinecarbonyl]formate I-45b (12.8 g, 95.98%) as an off-white solid. LCMS: m / z (ESI), [M+NH 4 + = 288.05.

[0320]

[0310] Step 2: In a 40 mL vial, ester I-45b (5000 mg, 18.36 mmol, 1 equiv) in dioxane (100 mL) at room temperature, Pd 2 (dba) 3 (840.8 mg, 0.92 mmol, 0.05 equiv), XPhos (1750.9 mg, 3.67 mmol, 0.2 equiv), (tert-butoxy)lithium (3528.2 mg, 44.06 mmol, 2.4 equiv) and 3-bromo-5-methoxypyridine (3452.8 mg, 18.36 mmol, 1.0 equiv) were added. The resulting mixture was stirred at 110 °C for 20 min under a nitrogen atmosphere. The reaction was quenched with saturated NH 4 Cl (aqueous solution) at 0 °C. The resulting mixture was quenched with water (100 mL) and extracted with ethyl acetate (3 × 100 mL). The combined organic layers were dried over anhydrous Na 2 SO 4 and filtered, and evaporated to give a crude solid. The residue was purified by CH 2 Cl 2 ​Purified by preparative TLC using dichloromethane / methanol (45:1, volume / volume) to obtain 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.

[0321]

[0311] Step 3: In a 50 mL round-bottom flask, enoate I-45c (1.5 g, 7.76 mmol, 1 equiv) in DMF (3 mL) at room temperature, triethylamine (2.36 g, 23.29 mmol, 3 equiv) and 1-methylpiperazine (777.65 mg, 7.76 mmol, 1 equiv) were added. The resulting mixture was stirred at room temperature for 1.5 h under a nitrogen atmosphere. The resulting mixture was concentrated under vacuum. The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, methanol in water, 0% to 100% concentration gradient in 30 min; detector, UV 254 nm. The resulting mixture was concentrated under vacuum to obtain methyl 2-(5-methoxypyridin-3-yl)-3-(4-methylpiperazin-1-yl)propanoate I-45d (900 mg, 39.51%) as a white oil. LCMS: m / z (ESI), [M+H] + = 294.10.

[0322]

[0312] Step 4: In a 50 mL round-bottom flask, methyl piperazine I-45d (900 mg, 3.07 mmol, 1 equiv) in water (2 mL) at room temperature, THF (2 mL) and LiOH·H 2 O (257.45 mg, 6.14 mmol, 2 equiv) were added. The resulting mixture was stirred at room temperature for 2 h. The mixture was acidified to pH 5 with HCl (aqueous solution). The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, methanol in water, 0% to 100% concentration gradient in 30 min; detector, UV 254 nm. The resulting mixture was concentrated under vacuum to obtain 2-(5-methoxypyridin-3-yl)-3-(4-methylpiperazin-1-yl)propanoic acid I-45 (330 mg, 38.51%) as a white solid. LCMS: m / z (ESI), [M+H]+ = 280.10 1 H NMR (DMSO-d 6 , 400 MHz) δ 2.70 (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). Synthesis of Intermediate I-46

[0323]

Chemical Structure

[0324]

[0313] In a 20 mL vial, at room temperature, tert-butyl N-(3-oxo-3-phenylpropyl)carbamate I-46a (400 mg, 1.60 mmol, 1 equivalent), sodium cyanoborohydride (403.30 mg, 6.42 mmol, 4 equivalents), and NH 4 OAc (1.24 g, 16.04 mmol, 10 equivalents) were added to methanol (20.00 mL). The resulting mixture was stirred at 60 °C overnight under a nitrogen atmosphere. The mixture was basified to pH 9 with NaOH (2 mol / L). The resulting mixture was extracted with ethyl acetate (3 × 100 mL). The combined organic layers were dried over anhydrous Na 2 SO 4 , filtered, and concentrated under reduced pressure. The residue was purified by preparative TLC using CH 2 Cl 2 / methanol (15:1, v / v) to obtain tert-butyl N-(3-amino-3-phenylpropyl)carbamate I-46 (250 mg, 62.24%) as a yellow oil. LCMS: m / z (ESI), [M + H] + = 251.15 1 H NMR (DMSO-d 6 , 400 MHz) δ 1.37 (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). Synthesis of Intermediate I-47

[0325] [Chemical Formula]

[0326]

[0314] Step 1: To a stirred mixture of N-methoxy-N,1-dimethylpiperidine-4-carboxamide I-47a (1.0 g, 5.37 mmol, 1 equiv) in THF (10 mL) was added dropwise bromo-(3-methoxyphenyl)magnesium (1.16 g, 5.48 mmol, 1.02 equiv) at 0 °C under an air atmosphere. The resulting mixture was stirred overnight at room temperature under an air atmosphere. After the reaction, the reaction was quenched with saturated NH 4 Cl (aqueous solution) (40 mL) at 0 °C. The resulting mixture was extracted with ethyl acetate (3 × 40 mL). The combined organic layers were washed with water (3 × 40 mL) and dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica column chromatography eluting with CH 2 Cl 2 / methanol (50:1, v / v) 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 H NMR (CDCl 3 , 400 MHz) δ 1.90 (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).

[0315] Step 2: To a stirred mixture of methylpiperidine I-47b (880 mg, 3.77 mmol, 1 equiv) in methanol (10 mL) was added NaBH 3 CN (948.08 mg, 15.09 mmol, 4 equiv) and CH 3 COONH 4(3488.89 mg, 45.26 mmol, 12 eq) was added portionwise. The resulting mixture was stirred overnight at 70 °C under an air atmosphere. After cooling to room temperature, the reaction was quenched at 0 °C by the addition of saturated NH 4 Cl (aqueous solution) (30 mL). The resulting mixture was extracted with CH 2 Cl 2 (3 × 40 mL). The combined organic layers were washed with water (3 × 10 mL) and dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, MeCN in water, gradient from 10% to 50% in 10 min; detector, UV 254 nm, held at 40% for 25 min to give 1-(3-methoxyphenyl)-1-(1-methylpiperidin-4-yl)methanamine I-47 (340 mg, 38.47%) as a yellow solid. LCMS: m / z (ESI), [M+H] + = 235.15.

[0327] Synthesis of Intermediate I-48

[0328]

Chemical Structure

[0329]

[0316] Step 1: In a 40 mL vial, N-methoxy-N,1-dimethylpiperidine-4-carboxamide I-47a (2 g, 10.74 mmol, 1 eq) and bromo(cyclohexyl)magnesium (21.48 mL, 21.48 mmol, 2 eq) in THF (10.00 mL) were added at room temperature. The resulting mixture was stirred overnight at room temperature under a nitrogen atmosphere. The reaction was quenched at room temperature with water. The resulting mixture was extracted with ethyl acetate (3 × 20 mL). The combined organic layers were concentrated under reduced pressure. The residue was taken up in CH 2 Cl 2Purified by silica column chromatography eluting with / methanol (20:1, volume / volume) 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-d 6 , 400 MHz) δ 1.08 - 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).

[0317] Step 2: In a 20 mL vial, at room temperature, methylpiperidine I-48b (260 mg, 1.24 mmol, 1 equivalent) in methanol (5 mL), NH 4 OAc (1.34 g, 17.39 mmol, 14 equivalents) and sodium cyanoborohydride (312.21 mg, 4.97 mmol, 4 equivalents) were added. The resulting mixture was stirred at 60 °C overnight under a nitrogen atmosphere. The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, water in MEOH, 0% to 100% concentration gradient in 30 minutes; detector, UV254 nm. The resulting mixture was concentrated under vacuum to give 1-cyclohexyl-1-(1-methylpiperidin-4-yl)methanamine I-48 (180 mg, 68.89%) as a brown oil. LCMS: m / z (ESI), [M+H] + = 211.20. 1 H NMR (DMSO-d 6 , 400 MHz) δ 0.96 (1H, qd), 1.04 - 1.12 (1H, m), 1.17 (4H, ddt), 1.22 - 1.34 (3H, 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). Synthesis of Intermediate I-49

[0330] [Chemical]

[0331]

[0318] Step 1: In a 250 mL round-bottom flask, at room temperature, 3-bromo-7-nitro-1H-indole I-49a (20 g, 82.97 mmol, 1 equivalent), tert-butyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazole-1-carboxylate (26.85 g, 91.27 mmol, 1.1 equivalents), K 2 CO 3 (34.40 g, 248.92 mmol, 3 equivalents), Pd(dppf)Cl 2 (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. The resulting mixture was stirred at 80 °C for 2 hours under a nitrogen atmosphere. The residue was purified by trituration with water (30 mL). This gave a crude solid (26 g), and the crude solid was triturated with methanol (50 mL), thereby obtaining tert-butyl 4-(7-nitro-1H-indol-3-yl)pyrazole-1-carboxylate I-49b (17 g, 39.31%) as a yellowish-brown solid. LCMS: m / z (ESI), [M - Boc] + = 229.05.

[0332]

[0319] Step 2: In a 100 mL round-bottom flask, at 0 °C, nitro-indole I-49b (2.5 g, 7.61 mmol, 1 equivalent) and Zn (3.98 g, 60.91 mmol, 8 equivalents) were added. A solution of NH 4 Cl (1.79 g, 33.51 mmol, 5 equivalents) in water (10 mL) was added to the above mixture at 0 °C. The resulting mixture was stirred at 0 °C for 1 hour under a nitrogen atmosphere. The resulting mixture was filtered, and the filtered cake was washed with ethyl acetate (2 × 20 mL). The filtrate was diluted with water (50 mL), the residue was extracted with EA (2 × 50 mL), and the combined organic layers were dried over anhydrous Na 2 SO 4 and filtered and evaporated to obtain a crude solid. The residue was CH2 Cl 2 Purified by preparative TLC using dichloromethane / 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.

[0333] Synthesis of Intermediate I-50

[0334]

Chemical Structure

[0335]

[0320] Step 1: 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-dioxaborolan-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)Cl 2 (52.42 mg, 64.19 μmol) in dioxane (5.00 mL) and water (1.00 mL) were stirred at 90 °C for 16 h under nitrogen. The solvent was removed under reduced pressure. The residue was purified by silica column chromatography eluting with a gradient of 0 to 10% methanol in DCM. The pure fractions were 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.

[0336]

[0321] 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 h. The reaction mixture was filtered through celite, washed with ethanol (3 × 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 brownish solid. LCMS: m / z (ESI), [M+H] + = 329.3.

[0337] Synthesis of Intermediate I-51

[0338]

Chemical formula

[0339]

[0322] Step 1: Methyl 1H-indole-7-carboxylate I-51a (5 g, 28.54 mmol, 1 equiv) and DCE (50 mL) were added to a 50 mL three-necked round-bottom flask at -10 °C. To the stirred mixture, NBS (6.10 g, 34.25 mmol, 1.2 equiv) was added portionwise at -10 °C under a nitrogen atmosphere. The resulting mixture was quenched with water (80 mL) and extracted with CH 2 Cl 2 (3 × 50 mL). The combined organic layers were dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica column chromatography eluting with PE / EA (5:1, v / v) 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 (400 MHz, DMSO-d6 ) δ 3.95 (3H, s), 7.27 (1H, t), 7.60 (1H, d), 7.74 (1H, dt), 7.88 (1H, dd), 11.51 (1H, s).

[0323] Step 2: Bromide I-51b (2 g, 7.87 mmol, 1 equivalent), Pd(dppf)Cl 2 ·CH 2 Cl 2 (0.64 g, 0.79 mmol, 0.1 equivalent), K 2 CO 3 (3.26 g, 23.61 mmol, 3 equivalents) and 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (1.53 g, 7.87 mmol, 1 equivalent) in 1,4-dioxane (4 mL), water (1 mL) was stirred at 80 °C for 12 h under a nitrogen atmosphere. The resulting mixture was diluted with water (30 mL) and extracted with ethyl acetate (3 × 30 mL), dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica column chromatography eluting with CH 2 Cl 2 / methanol (5:1, v / v) to give methyl 3-[1-(tert-butoxycarbonyl)pyrazol-4-yl]-1H-indole-7-carboxylate I-51c (810 mg, 19.41%) as a yellow oil. LCMS: m / z (ESI), [M+H] + = 242.00.

[0340]

[0324] Step 3: In a 50 mL round-bottom flask, ester I-51c (390 mg, 1.14 mmol, 1 equivalent), LiOH (136.81 mg, 5.71 mmol, 5 equivalents), THF (2.5 mL) and water (2.5 mL) were added at room temperature. The resulting mixture was stirred at room temperature for 8 hours. The mixture was acidified to pH 5 with HCl (1 M). The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, methanol in water, concentration gradient from 0% to 20% in 30 minutes; detector, UV 254 nm. Thereby, 3-(1H-pyrazol-4-yl)-1H-indole-7-carboxylic acid I-51 (60 mg, 13.98%) was obtained as a yellow solid. LCMS: m / z (ESI), [M+H] + = 242.10.

[0341] Synthesis of Intermediate I-52

[0342]

Chemical Structure

[0343]

[0325] 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), (Boc) 2 O (3576.56 mg, 16.39 mmol, 2 equivalents), TEA (2487.46 mg, 24.58 mmol, 3 equivalents) were added. The resulting mixture was stirred at room temperature for 4 hours under an air atmosphere. After the reaction, the resulting mixture was concentrated under vacuum and decomposed with DMF (4 mL). The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, MeOH in water, concentration gradient from 10% to 100% in 10 minutes; detector, UV 254 nm, held at 90% for 30 minutes to obtain methyl 2-(3-bromophenyl)-2-[(tert-butoxycarbonyl)amino]acetate I-52b (1.65 g, yield 58.50%) as a white solid. LCMS: m / z (ESI), [M - Boc] + = 243.95.

[0344]

[0326] Step 2: To a mixture of bromide I-52b (1.2 g, 3.49 mmol, 1 equiv) in DMF (4.0 mL) was added Zn(CN) 2 (417.55 mg, 3.56 mmol, 1.02 equiv), Pd(PPh 3 ) 4 (402.87 mg, 0.35 mmol, 0.1 equiv). The resulting mixture was stirred at 70 °C for 2 h under a nitrogen atmosphere. After cooling to room temperature, the resulting mixture was diluted with water (40 mL). The resulting mixture was extracted with EtOAc (3 × 40 mL). The combined organic layers were washed with water (3 × 10 mL) and dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC using PE / EA (3:1, v / v) to give methyl 2-[(tert-butoxycarbonyl)amino]-2-(3-cyanophenyl)acetate I-52c (990 mg, yield 97.81%) as a white solid. LCMS: m / z (ESI), [M - Boc + H] + = 191.00. 1 H NMR (DMSO-d 6 , 400 MHz) δ 1.40 (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).

[0327] Step 3: To a mixture of ester I-52c (990 mg, 3.41 mmol, 1 equiv) in THF (4.0 mL) was added LiOH·H in water (1.0 mL) 2O (572.34 mg, 13.64 mmol, 4 eq) was added. The resulting mixture was stirred at room temperature for 6 h under a nitrogen atmosphere. After the reaction, the resulting mixture was concentrated under vacuum and diluted with DMF (2 mL). The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, MeCN in water, concentration gradient from 10% to 50% in 10 min; detector, UV 254 nm, held at 25% for 25 min. After evaporation of water, [(tert-butoxycarbonyl)amino](3-cyanophenyl)acetic acid I-52 (540 mg, yield 57.31%) was obtained as a white solid. LCMS: m / z (ESI), [M+H] + = 277.00.

[0345] Synthesis of Intermediate I-53

[0346]

Chemical Structure

[0347]

[0328] Step 1: To a stirred solution of tert-butyl 4-bromobenzoate (20.11 g, 78.21 mmol, 1.55 eq) and methyl cyanoacetate (5 g, 50.46 mmol, 1.00 eq) in dioxane (40 mL), t-BuOK (14.16 g, 126.19 mmol, 2.50 eq) and Pd(OAc) 2 (1.13 g, 5.03 mmol, 0.10 eq), dppf (5.57 g, 10.08 mmol, 0.20 eq) were added under an air atmosphere at room temperature. The resulting mixture was stirred at 70 °C for 3 h under a nitrogen atmosphere. The reaction was quenched at room temperature by the addition of water (100 mL). The resulting mixture was extracted with EtOAc (3 × 50 mL). The combined organic layers were dried over anhydrous Na 2 SO 4 4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica column chromatography eluting with PE / EA (1:1, v / v) to give tert-butyl 4-(1-cyano-2-methoxy-2-oxoethyl)benzoate I-53b (3.77 g, yield 27.14%) as a yellow oil. 11H NMR: (DMSO-d 6 , 400 MHz) δ 1.55 (9H, s), 3.72 (3H, d), 5.84 (1H, s), 7.54 - 7.61 (2H, m), 7.94 - 8.02 (2H, m).

[0329] Step 2: Nitrile I-53b (3.77 g, 13.69 mmol, 1 equiv) and CoCl 2 ·6H 2 O (9.77 g, 41.08 mmol, 3 equiv) and (Boc) 2 O (8.97 g, 41.08 mmol, 3 equiv) were added portionwise to a stirred solution in MeOH (50 mL) at 0 °C with NaBH 4 (4.14 g, 109.55 mmol, 8 equiv). The resulting mixture was stirred at room temperature overnight. The reaction was quenched at room temperature by the addition of ice / water (50 mL). The precipitated solid was filtered off and washed with DCM (3 × 20 mL). The resulting mixture was extracted with CH 2 Cl 2 (3 × 50 mL). The combined organic layers were dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica column chromatography eluting with PE / EA (1:1, v / v) to give tert-butyl 4-{3-[(tert-butoxycarbonyl)amino]-1-methoxy-1-oxopropan-2-yl}benzoate I-53c (2947 mg, 56.71% yield) as a yellow oil. LCMS: m / z (ESI), [M+H] + = 380.45. 1 1H NMR: (CD 3 OD, 400 MHz) δ 1.41 (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).

[0330] Step 3: Ester I-53c (2937 mg, 7.74 mmol, 1 equiv) in MeOH (20 mL) and H 2To the stirred solution in O (10 mL), LiOH (741.50 mg, 30.96 mmol, 4 equiv) was added at room temperature under an air atmosphere. The resulting mixture was stirred at room temperature for 3 h under an air atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, MeCN in water, 90% to 100% concentration gradient over 10 min; detector, UV 254 nm. Thereby, 3-[(tert-butoxycarbonyl)amino]-2-[4-(tert-butoxycarbonyl)phenyl]propanoic acid I-53 (2.5 g, yield 88.39%) was obtained as a white solid. LCMS: m / z (ESI), [M+H] + = 366.20. 1 H NMR: (CD 3 OD, 400 MHz) δ 1.41 (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). Synthesis of Intermediate I-54

[0348]

Chemical Structure

[0349]

[0331] Step 1: To a solution of methyl 3-[(tert-butoxycarbonyl)amino]-2-(3-cyanophenyl)propanoate I-3d (200 mg, 0.66 mmol, 1 equiv) and CoCl 2 ·6H 2 O (469.05 mg, 1.97 mmol, 3 equiv) in MeOH (10 mL), Boc 2 O (430.26 mg, 1.97 mmol, 3 equiv) was added at 0 °C under a nitrogen atmosphere. To the mixture, NaBH 4(124.30 mg, 3.29 mmol, 5 eq) was added. The reaction mixture was stirred at 0 °C for 2 h. The resulting mixture was filtered, and the filter cake was washed with MeOH (3 × 20 mL). The filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC using PE:EA (2:1, v / v) to give methyl 3-[(tert-butoxycarbonyl)amino]-2-(3-{[(tert-butoxycarbonyl)amino]methyl}phenyl)-propanoate I-54a (180 mg, yield 47.94%) as a white solid. LCMS: m / z (ESI), [M+H] + = 409.10.

[0350]

[0332] Step 2: To a solution of ester I-54a (180 mg, 0.44 mmol, 1 eq) in THF (10 mL) and H 2 O (5 mL), LiOH·H 2 O (36.98 mg, 0.88 mmol, 2 eq) was added at room temperature under a nitrogen atmosphere. The mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, ACN in water, 10% to 50% concentration gradient in 10 min; detector, UV220 nm. Thereby, 3-[(tert-butoxycarbonyl)amino]-2-(3-{[(tert-butoxycarbonyl)amino]methyl}phenyl)-propanoic acid I-54 (159 mg, yield 91.27%) was obtained as a white solid. LCMS: m / z (ESI), [M+H] + = 395.10.

[0351] Synthesis of Intermediate I-55

[0352]

Chemical Structure

[0353]

[0333] Step 1: 1-(Benzyloxy)-4-bromobenzene (8232.15 mg, 31.29 mmol, 1.55 eq), t-BuOK (5662.23 mg, 50.46 mmol, 2.5 eq) and Pd(OAc) 2(453.15 mg, 2.02 mmol, 0.1 eq) of dioxane (50.00 mL, 590.18 mmol) was added dropwise to the stirred mixture under a nitrogen atmosphere at room temperature with dppf (2229.81 mg, 4.04 mmol, 0.2 eq) and methyl cyanoacetate (2 g, 20.18 mmol, 1 eq). The resulting mixture was stirred at 70 °C for 3 h under a nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica column chromatography eluting with PE / EA (4:1, v / v) to give methyl 2-[4-(benzyloxy)phenyl]-2-cyanoacetate I-55b (1.955 g, yield 34.43%) as a yellow solid. 1 H-NMR: (DMSO-d 6 , 400 MHz) δ 3.72 (3H, s), 5.13 (2H, s), 5.59 (1H, s), 7.09 (2H, m), 7.40 (7H, m).

[0334] Step 2: To a stirred mixture of nitrile I-55b (500 mg, 1.78 mmol, 1 eq) and CoCl 2 ·6H 2 O (1268.63 mg, 5.33 mmol, 3 eq) in MeOH (2 mL) was added portionwise Boc 2 O (1163.74 mg, 5.33 mmol, 3 eq) and NaBH 4 (168.10 mg, 4.44 mmol, 2.5 eq) at 0 °C under an air atmosphere. The resulting mixture was stirred at room temperature for 1 h under an air atmosphere. The reaction was quenched with water at room temperature. The resulting mixture was filtered and the filter cake was washed with MeOH (2 × 4 mL). The filtrate was concentrated under reduced pressure. The residue was purified by silica column chromatography eluting with PE / EA (3:1, v / v) to give methyl 2-[4-(benzyloxy)phenyl]-3-[(tert-butoxycarbonyl)amino]propanoate I-55c (371 mg, yield 54.15%) as a white solid. LCMS: m / z (ESI), [M - t Bu] + = 330.15.

[0354]

[0335] Step 3: Ester I-55c (371 mg, 0.96 mmol, 1 equivalent) and LiOH·H 2 O (121.16 mg, 2.89 mmol, 3 equivalents) in THF (4 mL) and H 2 O (1 mL) was stirred at room temperature for 2 hours under an air atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, MeCN in water, concentration gradient from 10% to 50% in 50 minutes; detector, UV254 nm. This gave 2-[4-(benzyloxy)phenyl]-3-[(tert-butoxycarbonyl)amino]propanoic acid I-55 (150 mg, yield 41.96%) as a yellow solid. LCMS: m / z (ESI), [M-H] - = 370.10. 1 H NMR: (DMSO-d 6 , 400 MHz) δ 1.34 (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). Synthesis of Intermediate I-56

[0355]

Chemical formula

[0356]

[0336] Step 1: In a 50 mL round-bottom flask, cyclohexaneacetic acid I-56a (2500 mg, 17.58 mmol, 1.0 equivalent) and SOCl 2(1.5 mL, 20.68 mmol) was added. The resulting mixture was stirred at 80 °C for 1 hour under a nitrogen atmosphere. The reaction mixture was cooled to room temperature. Phosphorus tribromide (4.26 mL, 44.83 mmol, 2.55 eq) and bromine (3512.00 mg, 21.98 mmol, 1.25 eq) were added to the above mixture at room temperature. The resulting mixture was stirred at 80 °C for an additional 2 hours. The mixture was cooled to room temperature. Then MeOH (2.5 mL) was added to the above solution. The resulting mixture was stirred at 70 °C for 2 hours under a nitrogen atmosphere. The mixture was cooled to room temperature. The reaction was quenched with saturated NaS 2 O 3 (aqueous solution) (50 mL). The resulting mixture was extracted with DCM (2 × 50 mL). The combined organic layers were dried over anhydrous Na 2 SO 4 and filtered and evaporated to give a crude solid. The crude solid was purified by silica column chromatography eluting with PE / EA (5:1, v / v) to give methyl 2-bromo-2-cyclohexylacetate I-56b (2000 mg, 40.25% yield) as an off-white oil. 1 H NMR: (CDCl 3 , 400 MHz) δ 0.90 - 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).

[0337] Step 2: In a 40 mL vial, bromide I-56b (2000 mg, 8.52 mmol, 1 eq) in ACN (20.0 mL) at room temperature, 1-methyl-piperazine (1022.44 mg, 10.21 mmol, 1.2 eq), K 2 CO 3 (3526.84 mg, 25.52 mmol, 3.0 eq) were added. The resulting mixture was stirred at 80 °C for 12 hours under a nitrogen atmosphere. The mixture was cooled to room temperature. The resulting mixture was concentrated under reduced pressure. The residue was taken up in CH 2 Cl 2Purified by silica column chromatography eluting with n-hexane / EtOAc / MeOH (20:1, volume / volume) to give methyl 2-cyclohexyl-2-(4-methylpiperazin-1-yl)acetate I-56c (950 mg, yield 24.23%) as an off-white solid. LCMS: m / z (ESI), [M+H] + = 255.15. 1 H NMR: (CDCl 3 , 400 MHz) δ 0.83 - 0.99 (3H, m), 1.15 - 1.32 (4H, m), 1.53 (1H, d), 1.78 (4H, dtd), 1.95 (1H, d), 2.30 (3H, s), 2.53 (6H, dd), 2.64 (2H, dt), 2.89 (1H, d), 3.68 (3H, s).

[0338] Step 3: A solution of methyl piperazine I-56c (590 mg, 2.32 mmol, 1 equiv) and LiOH·H 2 O (194.66 mg, 4.64 mmol, 2 equiv) in MeOH (3.0 mL) and H 2 O (3.0 mL) was stirred at 70 °C for 2 h under a nitrogen atmosphere. 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 reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, MeOH in water, 10% to 50% concentration gradient over 30 min; detector, UV 220 nm. This gave cyclohexyl(4-methylpiperazin-1-yl)acetic acid I-56 (240 mg, yield 35.8%) as an off-white solid. LCMS: m / z (ESI), [M+H] + = 241.15.

[0357] Synthesis of Intermediate I-57

[0358]

Chemical Structure

[0359]

[0339] Step 1: To a stirred mixture of methyl 3-[(tert-butoxycarbonyl)amino]-2-(4-cyanophenyl)propanoate I-14c (740 mg, 2.43 mmol, 1 equiv) and CoCl 2 ·6H 2 O (1156.98 mg, 4.86 mmol, 2 equiv) in MeOH (10 mL), NaBH 4 (919.81 mg, 24.31 mmol, 10 equiv) was added portionwise at 0 °C under an air atmosphere. The resulting mixture was stirred at room temperature for 5 h under an air atmosphere. 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 × 10 mL). The filtrate was extracted with EtOAc (3 × 20 mL). The combined organic layers were dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure to give the crude product. The crude product was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, MeCN in water, 10% to 50% concentration gradient over 10 min; detector, UV 254 nm, held at 15% for 20 min. After evaporation of the solvent under reduced pressure, methyl 2-[4-(aminomethyl)phenyl]-3-[(tert-butoxycarbonyl)amino]propanoate I-57b (300 mg, yield 40.01%) was obtained as a white solid. LCMS: m / z (ESI), [2M + H] + = 617.40.

[0360]

[0340] Step 2: To a stirred mixture of amine I-57b (200 mg, 0.649 mmol, 1 equiv) and acetyl chloride (51.93 mg, 0.66 mmol, 1.02 equiv) in DCM (3.0 mL), Et 3 N (196.89 mg, 1.95 mmol, 3 equiv) was added portionwise at 0 °C under an air atmosphere. The resulting mixture was stirred at room temperature for 3 h under an air atmosphere. After the reaction, the resulting mixture was diluted with water (10 mL). The resulting mixture was extracted with CH 2 Cl 2 (3 × 20 mL). The combined organic layers were dried over anhydrous Na 2 SO 4It was dehydrated. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC using CH 2 Cl 2 / MeOH (30:1, v / v) to obtain methyl 3-[(tert-butoxycarbonyl)amino]-2-[4-(acetamidomethyl)phenyl]propanoate I-57c (100 mg, yield 44.00%) as a white solid. LCMS: m / z (ESI), [M-Boc] + =251.15.

[0361]

[0341] Step 3: To a stirred mixture of ester I-57c (150 mg, 0.43 mmol, 1 equiv) in MeOH (4.0 mL), LiOH·H 2 O (71.85 mg, 1.71 mmol, 4 equiv) in water (1.0 mL) was added dropwise at room temperature under an air atmosphere. The resulting mixture was stirred overnight at 60 °C under an 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 reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, MeCN in water, 10% to 50% concentration gradient over 10 min; detector, UV 254 nm, held at 26% for 30 min. After evaporating the solvent under reduced pressure, 3-[(tert-butoxycarbonyl)amino]-2-[4-(acetamidomethyl)phenyl]propanoic acid I-57 (92 mg, yield 63.89%) was obtained as a white solid. LCMS: m / z (ESI), [M+Na] + =359.00.

[0362] Synthesis of Intermediate I-58

[0363]

Chemical Structure

[0364]

[0342] Step 1: In a 40 mL vial, methyl 3-[(tert-butoxycarbonyl)amino]-2-(4-{[(tert-butyldimethylsilyl)oxy]methyl}phenyl)propanoate I-1e (1.5 g, 3.54 mmol, 1 equiv), TBAF (2.78 g, 10.62 mmol, 3 equiv), and THF (25 mL) were added at room temperature. The resulting mixture was stirred at room temperature for 3 h under an air atmosphere. The reaction was quenched at room temperature by the addition of water (50 mL). The aqueous layer was extracted with EtOAc (3 × 50 mL). The resulting mixture was concentrated under reduced pressure. The residue was purified by preparative TLC using CH 2 Cl 2 / EA (1:1, v / v) to give methyl 3-[(tert-butoxycarbonyl)amino]-2-[4-(hydroxymethyl)phenyl]propanoate I-58a (975 mg, yield 89.01%) as a yellow oil. LCMS: m / z (ESI), [M+H] + = 310.20. 1 H NMR (CD 3 OD, 400 MHz) δ 1.42 (9H, s), 3.39 (1H, m), 3.64 (5H, m), 4.60 (2H, s), 5.51 (1H, s), 7.31 (4H, m).

[0343] Step 2: In a 40 mL vial, methyl 3-[(tert-butoxycarbonyl)amino]-2-[4-(hydroxymethyl)phenyl]propanoate (650 mg, 2.10 mmol, 1 equiv), N-methylcarbamoyl chloride (432.24 mg, 4.62 mmol, 2.2 equiv), TEA (1.06 g, 10.51 mmol, 5 equiv), and DCM (15 mL) were added at room temperature. The resulting mixture was stirred at room temperature for 3 h under an air atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was purified by preparative TLC using CH 2 Cl 2Purified by preparative TLC using / MeOH (40:1, volume / volume) to obtain methyl 3-[(tert-butoxycarbonyl)amino]-2-(4-{[(methylcarbamoyl)oxy]methyl}phenyl)-propanoate I-58b (570 mg, yield 74.04%) as a yellow oil. LCMS: m / z (ESI), [M+H] + = 367.05.

[0365]

[0344] Step 3: In a 100 mL round-bottom flask, ester I-58b (570 mg, 1.56 mmol, 1.00 equivalent), LiOH·H2O (163.18 mg, 3.89 mmol, 2.5 equivalents), MeOH (4 mL), and water (2 mL) were added at room temperature. The resulting mixture was stirred at room temperature for 16 h under an air atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, MeOH in water (0.1% FA), 0% to 25% concentration gradient in 20 min; detector, UV 254 nm. The resulting mixture was concentrated under reduced pressure to obtain 3-[(tert-butoxycarbonyl)amino]-2-(4-{[(methylcarbamoyl)-oxy]methyl}phenyl)propanoic acid I-58 (520 mg, yield 94.86%) as an off-white solid. LCMS: m / z (ESI), [M+H - Boc] + = 253.15.

[0366] Synthesis of Intermediate I-59

[0367]

Chemical Structure

[0368]

[0345] Step 1: In a 50 mL round-bottom flask, at 0 °C, methyl 2-(3-methoxyphenyl)acetate I-59a (3 g, 16.65 mmol, 1 equiv), t-BuOK (3.74 g, 33.30 mmol, 2 equiv) and tert-butyl 4-iodopiperidine-1-carboxylate (6.73 g, 21.642 mmol, 1.3 equiv) in DMF (10 mL) were added. The resulting mixture was stirred at room temperature for 2 h. The residue was purified by silica column chromatography eluting with PE / EA (25:1, v / v) 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-d 6 , 400 MHz) δ 0.83 -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).

[0346] Step 2: In a 50 mL round-bottom flask, at room temperature, piperidine I-59b (2.0 g, 5.50 mmol, 1 equiv) and TFA (1.5 mL) in DCM (3 mL) were added. The resulting mixture was stirred at room temperature for 1.5 h. The mixture was basified to pH 9 with saturated NaHCO 3 (aqueous solution). The resulting mixture was extracted with DCM (3 × 50 mL). The combined organic layers were concentrated 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-d 6, 400 MHz) δ 1.08 - 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).

[0347] Step 3: In a 50 mL round bottom flask, at room temperature, piperidine I-59c (1.3 g, 4.94 mmol, 1 equivalent) in MeOH (2.5 mL), DIEA (3.19 g, 24.69 mmol, 5.0 equivalents), sodium cyanoborohydride (620 mg, 9.87 mmol, 2 equivalents) and CH 2 O (296 mg, 9.88 mmol, 2 equivalents) were added. The resulting mixture was stirred at room temperature overnight. The reaction was quenched with water at room temperature. The resulting mixture was extracted with EtOAc (3 × 10 mL). The combined organic layers were concentrated under reduced pressure to give methyl piperidine I-59d (1.1 g, 80.34%) as a yellow oil. LCMS: m / z (ESI), [M + H] + = 278.25.

[0369]

[0348] Step 4: In a 20 mL vial, at room temperature, methyl piperidine I-59d (600 mg, 2.16 mmol, 1 equivalent) in THF (2 mL) and LiOH (104 mg, 4.33 mmol, 2 equivalents) in water (2 mL) were added. The resulting mixture was stirred at room temperature for 3 hours. The mixture was acidified to pH 5 with HCl (1 M). The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, MeOH in water, 0% to 30% concentration gradient over 40 minutes; detector, UV 254 nm to give acid I-59 (270 mg, 47.40%) as a white solid. LCMS: m / z (ESI), [M + H] + = 264.20. 1 H NMR (DMSO-d 6, 400 MHz) δ 0.95 - 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). Synthesis of Intermediate I-60

[0370] [Chemical Structure]

[0371]

[0349] Step 1: In a 20 mL vial, at room temperature, ester I-60a (2 g, 11.10 mmol, 1 equivalent), formaldehyde (0.67 g, 22.20 mmol, 2 equivalents), K 2 CO 3 (4.60 g, 33.30 mmol, 3 equivalents) and DMF (2 mL) were added. The resulting 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), washed with water (3 × 30 mL), and the combined organic layers were dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC using PE / EA (8:1, volume / volume) to obtain enoate I-60b (1.3 g, 60.88%) as a pale yellow oil. 1 H NMR (CD 3 OD, 400 MHz) δ 3.82 (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).

[0350] Step 2: In a 100 mL round-bottom flask, enolate I-60b (1.3 g, 6.76 mmol, 1 equiv) was added at room temperature, 1-methylpiperazine (2.71 g, 27.05 mmol, 4 equiv), TEA (2.05 g, 20.29 mmol, 3 equiv) and DMF (10 mL). The resulting mixture was stirred at room temperature for 1 hour. The resulting mixture was diluted with EtOAc (50 mL) and washed with 2×30 mL of water and 1×30 mL of saturated brine. The resulting mixture was concentrated under reduced pressure. This gave methylpiperidine I-60c (1.7 g, 81.33%) as a yellowish-brown oil. LCMS: m / z (ESI), [M+H ]+ =293.15.

[0372]

[0351] Step 3. In an 80 mL vial, methylpiperidine I-60c (1 g, 3.42 mmol, 1 equiv) was added at room temperature, LiOH (163.83 mg, 6.84 mmol, 2 equiv), THF (5 mL) and water (5 mL). The resulting mixture was stirred at room temperature for 1 hour. The mixture was acidified to pH 5 with HCl (1 M). The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, MeOH in water, concentration gradient from 10% to 50% in 30 minutes; detector, UV220 nm. This gave acid I-60 (497 mg, 51.16%) as a white solid. LCMS: m / z (ESI), [M+H] + =279.15.

[0373] Synthesis of Compounds Example 1 (racemate), Example 2 (enantiomer 1) and Example 3 (enantiomer 2) Preparation of N-(3-(1H-pyrazol-4-yl)-1H-indol-7-yl)-3-amino-2-(4-(hydroxymethyl)phenyl)propanamide

[0374]

Chem.

[0375]

[0352] Step 1: A mixture of acid I-1 (498.91 mg, 1.69 mmol, 1.2 eq), amine I-49 (420 mg, 1.41 mmol, 1 eq), HATU (802.92 mg, 2.11 mmol, 1.5 eq), and triethylamine (427.37 mg, 4.22 mmol, 3 eq) in DMF (5 mL) was stirred overnight at room temperature under a nitrogen atmosphere. The resulting mixture was diluted with ethyl acetate (100 mL). The residue was washed with saturated NaHCO 3 (1×80 mL), water (2×80 mL), and saturated NaCl (1×80 mL). The residue was purified by reverse flash chromatography using the following conditions: column, C18 silica; mobile phase, MeCN in water, gradient from 35% to 65% in 40 min; detector, UV254 nm to obtain amide C-1a (500 mg, 61.70%) as an off-white solid. LCMS: m / z (ESI), [M+H] + = 576.5. 1 H NMR (DMSO-d 6 , 400 MHz) δ 1.37 (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).

[0353] Step 2: Amide C-1a (250 mg) was purified by preparative HPLC using the following conditions: column, CHIRALPAK ID, 2×25 cm, 5 μm; mobile phase A: Hex (0.2% DEA), mobile phase B: EtOH; flow rate: 20 mL / min; concentration gradient: 50% B in 13 min; wavelength: 220 / 254 nm; sample solvent: EtOH) to obtain the enantiomeric amide C-1b (80 mg, 32.00%) as an off-white solid. t R = 7.48 min. LCMS: m / z (ESI), [M+H] + = 576.2. 1 H NMR (CD 31H NMR (CD3OD, 400 MHz) δ 1.44 (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 min, ee = 100% and enantiomer C-1c (85.3 mg, 32.52%) was obtained as an off-white solid. t R = 9.78 min. LCMS: m / z (ESI), [M+H] + = 576.2. 1 1H NMR (CD 3 3OD, 400 MHz) δ 1.44 (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 = 1.89 min, ee = 99.76%.

[0376]

[0354] Step 3: (Example 3, Enantiomer 2): A solution of enantiomer C-1c (33 mg, 57.33 μmol) in DCM (1.48 mL) was cooled to 0 °C in an ice / water bath and stirred. Then trifluoroacetic acid (261.45 mg, 2.29 mmol, 176.66 μL) was added dropwise to the reaction mixture. The resulting mixture was stirred for 0.5 h. The reaction solution was concentrated under reduced pressure. The residue was dissolved in water (6 mmol / L NH 4 4HCO 3) Purified by C18 flash chromatography column eluting with a concentration gradient of 0% to 60% MeCN. The pure fraction was lyophilized to obtain amine Example 3, enantiomer 2 (20.76 mg, 93% yield) as a white solid. LCMS: 3-minute chromatography (5 - 95% MeCN in water (0.02% TFA), waters Acquity UPLC BEH C18 1.7um, 2.1×50mm, 40 °C) with t R = 1.09 minutes, MS (ESI) m / z = 376.2 [M+H] + . 1 H NMR: (400 MHz, DMSO-d 6 ) δ 12.80 (s, 1H), 10.83 (s, 1H), 7.94 (s, 2H), 7.57 (d, J = 12.1 Hz, 2H), 7.49 - 7.31 (m, 3H), 7.29 (d, J = 7.9 Hz, 2H), 6.98 (t, J = 7.8 Hz, 1H), 5.14 (s, 1H), 4.46 (s, 2H), 3.92 - 3.84 (m, 1H), 2.89 (dd, J = 11.7, 5.0 Hz, 1H).

[0355] Using amide C1-b as the starting material, Example 2 (enantiomer 1) can be obtained according to the above procedure.

[0377]

[0356] Step 4: (Example 1, racemate): In a 100 mL round-bottom flask, amide C-1a (60 mg, 0.10 mmol, 1 equivalent) in DCM (3 mL) and TFA (1 mL) was added at 0 °C. The resulting mixture was stirred at 0 °C for 1 hour under a nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in methanol (3 mL). The mixture was basified to pH 9 with NH 3 (aqueous solution). The crude product was separated by preparative HPLC using the following conditions (column: Xselect CSH C18 OBD 30×150mm; mobile phase A: water (0.1% FA), mobile phase B: ACN; flow rate: 60 mL / min; concentration gradient: 5% B to 18% B in 7 minutes; wavelength: 254; 220 nm; t RPurified by preparative HPLC (column: XBridge Prep OBD C18 30×150mm, 5μm; mobile phase A: water (10 mmol / L NH4HCO3), mobile phase B: ACN; flow rate: 60 mL / min; concentration gradient: 55% B to 85% B in 7 minutes; wavelength: 220 nm; tR = 6.05) to obtain the racemic Example 1 (21.5 mg, 54.94%) as a white solid. LCMS: m / z (ESI), [M+H] + = 376.3. 1 H NMR (CD 3 OD, 400 MHz) δ 3.37 (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). Example 5 (enantiomer 1) and Example 6 (enantiomer 2) Preparation of 4-(1-((3-(1H-pyrazol-4-yl)-1H-indol-7-yl)amino)-3-amino-1-oxopropan-2-yl)benzyl 3,5-dimethylbenzoate

[0378] [Chemical formula]

[0379]

[0357] Step 1: In a 50 mL round-bottom flask, alcohol C-1a (146 mg, 0.25 mmol, 1 equivalent), 2,4-dimethylbenzoic acid (45.71 mg, 0.31 mmol, 1.2 equivalents), EDCI (72.93 mg, 0.38 mmol, 1.5 equivalents), and DMAP (30.98 mg, 0.25 mmol, 1 equivalent) in DMF (5 mL) were added at room temperature. The resulting mixture was stirred at room temperature under a nitrogen atmosphere for 4 hours. The product precipitated upon addition of water. The crude product (146 mg) was purified by preparative HPLC using the following conditions (column: XBridge Prep OBD C18 30×150mm, 5μm; mobile phase A: water (10 mmol / L NH 4 HCO 3 3), mobile phase B: ACN; flow rate: 60 mL / min; concentration gradient: 55% B to 85% B in 7 minutes; wavelength: 220 nm; t R=8.25 min) and purified to obtain racemic ester C-5a (130 mg, 72.42%) as an off-white solid. LCMS: m / z (ESI), [M+H] + =708.30. 1 H NMR (DMSO-d 6 , 400 MHz) δ 1.36 (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).

[0358] Step 2: Racemic ester C-5a (260 mg) was purified by preparative chiral HPLC using 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 ul mL) to obtain enantiomeric ester C-5c, SFC: t R =2.79 min, ee = 100% and enantiomeric ester C-5b, SFC: t R =3.567 min, ee = 100%.

[0380]

[0359] Step 3: Enantiomeric ester C-5c (110 mg, 0.16 mmol, 1 equiv) in DCM (3 mL) and TFA (1 mL) was added to a 100 mL vial at room temperature. The resulting mixture was stirred at room temperature for 30 min under a nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in methanol (3 mL). The residue was NH 3It was basified to pH 8 with an aqueous solution. The resulting mixture was stirred at 0 °C for 1 hour under a nitrogen atmosphere. The crude product (80 mg) was purified by preparative HPLC using the following conditions (column: XBridge Shield RP18 OBD 30×150 mm, 5 μm; mobile phase A: water (10 mmol / L NH 4 HCO 3 ), mobile phase B: ACN; flow rate: 60 mL / min; concentration gradient: 30% B to 60% B in 8 minutes; wavelength: 220 nm; t R = 7.58 minutes) to obtain Example 6, enantiomer 2 (28.8 mg, 36.51%) as a white solid. LCMS: m / z (ESI), [M+H] + = 508.25. 1 H NMR (CD 3 OD, 400 MHz) δ 2.35 (3H, s), 2.54 (3H, s), 3.08 (1H, dd), 3.46 (1H, dd), 4.01 (1H, dd), 5.33 (2H, s), 6.98 - 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%.

[0381]

[0360] In a 100 mL vial, enantiomeric ester C-5b (110 mg, 0.16 mmol, 1 equivalent) in DCM (3 mL) and TFA (1 mL) was added at room temperature. The resulting mixture was stirred at room temperature for 30 minutes under a nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in methanol (3 mL). The residue was basified to pH 8 with an NH 3 aqueous solution. The resulting mixture was stirred at 0 °C for 1 hour under a nitrogen atmosphere. The crude product was purified by preparative HPLC using the following conditions (column: XBridge Prep OBD C18 30×150 mm, 5 μm; mobile phase A: water (10 mmol / L NH 4 HCO 3 ), mobile phase B: ACN; flow rate: 60 mL / min; concentration gradient: 34% B to 54% B in 8 minutes; wavelength: 254 nm; tR = 7.83 min) for purification to give Example 5, enantiomer 1 (24.5 mg, 42.71%) as a white solid. LCMS: m / z (ESI), [M+H] + = 508.25 1 H NMR (CD 3 OD, 400 MHz) δ 2.35 (3H, s), 2.55 (3H, s), 3.04 (1H, dd), 3.43 (1H, dd), 3.96 (1H, dd), 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 min, ee = 100%

[0382] Example 7 (racemate) Preparation of N-(3-(1H-pyrazol-4-yl)-1H-indol-7-yl)-2-amino-2-phenylacetamide (racemate)

[0383]

Chemical Structure

[0384]

[0361] 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 h. The reaction mixture was filtered through Celite, washed with ethanol (50 mL × 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, yield 67%) as a brown solid. LCMS: 3-minute chromatography (3 min - 5 - 95% MeCN in water (0.02% TFA), Waters Acquity UPLC BEH C18 1.7um, 2.1 × 50 mm, 40 °C) with t R= 2.07 min, MS(ESI) m / z = 413.2 [M+H] + 。

[0385]

[0362] Step 2: 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) in NMP (20 mL) were stirred at room temperature for 15 minutes. Then DIPEA (4.37 g, 33.83 mmol, 5.89 mL) was added at room temperature. The resulting suspension was stirred at room temperature for 16 hours. The reaction mixture was diluted with 200 ml of water, then stirred at room temperature for 30 minutes, filtered, and washed with water (10 mL×2). The solid was collected and dried to obtain the amide racemate C-7a (1.60 g, 33% yield) as a brown solid. LCMS: 3-minute chromatography (3 minutes - 5 - 95% MeCN in water (0.02% TFA), Waters Acquity UPLC BEH C18 1.7um, 2.1×50 mm, 40 °C) with t R = 2.13 min, MS(ESI) m / z = 646.7 [M+H] + 。

[0386]

[0363] Step 3: Racemic C-7a (200 mg, 309.80 μmol), 4-bromo-1-tosyl-1H-pyrazole (139.95 mg, 464.70 μmol), K 2 CO 3 (128.45 mg, 929.41 μmol) and PdCl 2 dppf (25.30 mg, 30.98 μmol) in dioxane (3.00 mL) and water (0.60 mL) were stirred at 90 °C under nitrogen for 16 hours. The solvent was removed under reduced pressure. The residue was purified by silica flash chromatography column eluting with a concentration gradient of 0 to 40% ethyl acetate in petroleum ether. The pure fractions were evaporated to dryness to obtain the racemic amide C-7b (136 mg, 59% yield) as a brown solid. LCMS: 3-minute chromatography (3 minutes - 5 - 95% MeCN in water (0.02% TFA), Waters Acquity UPLC BEH C18 1.7um, 2.1×50 mm, 40 °C) with tR = 2.01 minutes, MS(ESI) m / z = 740.7 [M+H] + 。

[0387]

[0364] Step 4 and Step 5: Racemic amide C-7b (136 mg, 183.82 μmol) was placed in THF (0.60 mL) / MeOH (0.60 mL), and then 2N sodium hydroxide in water (73.53 mg, 1.84 mmol, 34.52 μL) was added to the reaction solution. The reaction mixture was stirred at room temperature for 3 hours. When the reaction was complete, the mixture was stirred and cooled to 0 °C in an ice / water bath, and then saturated NH 4 Cl aqueous solution (20 mL) was added, and the mixture was extracted with dichloromethane (10 mL × 3). The combined organic layers were washed with brine (50 mL × 1), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain crude racemic amide C-7c, which was dissolved in DCM (2.00 mL). The reaction solution 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 reaction solution. The resulting mixture was stirred at 0 °C for 2 hours. Then, the resulting mixture was concentrated under reduced pressure. The residue was purified by C18 flash chromatography eluting with a concentration gradient of 0% to 30% MeCN in water (0.02% TFA). The pure fractions were lyophilized to obtain racemic Example 7 (3.5 mg, 7.88 μmol, yield 4.28%, TFA salt) as a brown solid. LCMS: 3-minute chromatography (3 minutes - 5 to 95% MeCN in water (0.02% TFA), Waters Acquity UPLC BEH C18 1.7um, 2.1 × 50 mm, 40 °C) with t R = 0.93 minutes, MS(ESI) m / z = 332.4 [M+H] + 。 1 H NMR: (400 MHz, DMSO-d 6) δ 8.42 (s, 2H), 8.36 (d, J = 7.7 Hz, 1H), 8.29 - 8.25 (m, 1H), 8.16 (dd, J = 7.5, 1.9 Hz, 1H), 8.10 (d, J = 7.8 Hz, 1H), 7.96 - 7.84 (m, 5H), 7.60 (dd, J = 7.4, 1.0 Hz, 1H), 7.46 (t, J = 7.4 Hz, 1H), 6.35 (s, 1H). Example 8 (2R)-2-Amino-2-phenyl-N-[3-(1H-pyrazol-4-yl)-1H-indol-7-yl]acetamide Preparation

[0388]

Chemical formula

[0389]

[0365] Step 1: In an 80 mL vial, 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 equivalents), TCFH (225.71 mg, 0.80 mmol, 2 equivalents), NMI (165.12 mg, 2.01 mmol, 5 equivalents) and MeCN (5 mL) were added. The resulting mixture was stirred at room temperature for 8 hours. The resulting mixture was concentrated under reduced pressure. The residue was purified by preparative TLC using CH 2 Cl 2 / methanol (40:1, v / v) to obtain amide (R)-C-8 (160 mg, 83.39%) as a yellow solid. LCMS: m / z (ESI), [M+H- t Bu] + =476.20.

[0390]

[0366] Step 2: In a 50 mL round-bottom flask, at room temperature, amide (R)-C-8 (120 mg, 0.23 mmol, 1 equivalent), TFA (1 mL) and DCM (3 mL) were added. The resulting mixture was stirred at room temperature for 1 hour. The resulting mixture was CH 2 Cl 2It was diluted with (20 mL). The mixture was cooled to 0 °C. The mixture was basified to pH 9 with an aqueous solution of NH 3 The crude product (100 mg) was purified by preparative HPLC using the following conditions (column: XBridge Shield RP18 OBD 30×150 mm, 5 μm; mobile phase A: water (10 mmol / L NH 4 HCO 3 + 0.1% NH 3 aqueous solution), mobile phase B: ACN; flow rate: 60 mL / min; concentration gradient: from 17% B to 40% B in 8 minutes; wavelength: 220 nm; t R = 7.67 minutes). The crude product (60 mg) was purified by preparative chiral HPLC using 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; concentration gradient: from 45% B to 45% B in 16 minutes; wavelength: 220 / 254 nm; t R -1 = 8.97 minutes; t R -2 = 12.51 minutes; sample solvent: EtOH / DCM (1:1, v / v); injection volume: 0.7 mL). The crude product (32 mg) was purified by preparative HPLC using the following conditions (column: XBridge Shield RP18 OBD 30×150 mm, 5 μm; mobile phase A: water (10 mmol / L NH 4 HCO 3 + 0.1% NH 3 aqueous solution), mobile phase B: ACN; flow rate: 60 mL / min; concentration gradient: from 17% B to 40% B, 40% B in 8 minutes; 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 (CD 3 OD, 400 MHz) δ 4.76 (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: tR = 0.938 min, ee = 100%.

[0391] Example 9 Preparation of (2S)-2-Amino-2-phenyl-N-[3-(1H-pyrazol-4-yl)-1H-indol-7-yl]acetamide

[0392]

Chemical Structure

[0393]

[0367] Step 1: A solution of amine I-49 (150 mg, 0.50 mmol, 1 eq), (S)-[(tert-butoxycarbonyl)amino](phenyl)acetic acid (189.51 mg, 0.75 mmol, 1.5 eq), TCFH (211.60 mg, 0.75 mmol, 1.5 eq), and NMI (144.48 mg, 1.76 mmol, 3.5 eq) in MeCN (3 mL) was stirred at room temperature for 1 hour under a nitrogen atmosphere. The residue was purified by preparative TLC using CH 2 Cl 2 / methanol (22:1, v / v) to obtain amide (S)-C-9 (200 mg, 74.83%) as a yellowish-brown solid. LCMS: m / z (ESI), [M + H - t Bu] + = 476.25.

[0394]

[0368] Step 2: A solution of amide (S)-C-9 (160 mg, 0.27 mmol, 1 eq) and TFA (1.00 mL) in DCM (3 mL) was stirred at room temperature for 1 hour under a nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure. The resulting mixture was diluted with methanol (2 mL). The mixture was basified to pH 9 with aqueous ammonia at 0 °C and stirred for 30 minutes. The crude product (200 mg) was subjected to preparative HPLC using the following conditions (column: XBridge Shield RP18 OBD 30×150 mm, 5 μm; mobile phase A: water (10 mmol / L NH 4 HCO 3 + 0.1% NH 3Mobile phase A: aqueous solution; mobile phase B: ACN; flow rate: 60 mL / min; concentration gradient: from 17% B to 40% B in 8 minutes; wavelength: 220 nm; t R = 7.77 minutes) for purification to obtain (S)-Example 9 (46.6 mg, 20.74%) as an off-white solid. LCMS: m / z (ESI), [M+H] + = 332.15. 1 H NMR (CD 3 OD, 400 MHz) δ 4.75 (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%.

[0395] Example 10 (enantiomer 1) [4-[(1S)-1-(Aminomethyl)-2-oxo-2-[[3-(1H-pyrazol-4-yl)-1H-indol-7-yl]amino]ethyl]phenyl]methyl 6-nitrooxyhexanoate Preparation

[0396]

Chemical formula

[0397]

[0369] Step 1: 6-Bromohexanoic acid C-10a (500 mg, 2.56 mmol) and silver nitrate (566.09 mg, 3.33 mmol, 130.08 μL) in MeCN (12.5 mL), at 70 °C for 18 hours. Then the reaction solution was evaporated under reduced pressure. Then DCM (10 mL) was added. The suspension was stirred at room temperature for 15 minutes, filtered, and evaporated under reduced pressure to obtain 6-nitrooxyhexanoic acid C-10b (440 mg, yield 97%) as a brown oil. 1 H NMR: (400 MHz, CDCl 3) δ 11.18 (s, 1H), 4.45 (t, J = 6.6 Hz, 2H), 2.38 (t, J = 7.3 Hz, 2H), 1.83 - 1.63 (m, 4H), 1.56 - 1.40 (m, 2H).

[0370] Step 2: 6-Nitrooxyhexanoic acid C-10b (23.08 mg, 130.29 μmol) was dissolved in DCM (1.50 mL) at 0 °C in an ice / water bath. EDCI (24.98 mg, 130.29 μmol), DMAP (1.06 mg, 8.69 μmol) and enantiomeric alcohol C-1c (50 mg, 86.86 μmol) were added. The reaction mixture was stirred and slowly warmed to 25 °C over 16 h. The resulting mixture was diluted with water (50 mL) and extracted with ethyl acetate (20 mL × 3). The combined organic layers were washed with brine (60 mL × 1), dried over anhydrous sodium sulfate, filtered and evaporated under reduced pressure to give enantiomeric ester C-10c (62 mg, 97% yield) as a brownish solid. LCMS: 3-minute chromatography (5 - 95% MeCN in water (6 mmol / L NH 4 HCO 3 ) on a waters Acquity UPLC BEH C18 1.7um, 2.1×50mm, 40 °C) with t R = 2.07 min, MS (ESI) m / z = 735.2 [M+H] + .

[0398]

[0371] Step 3: A solution of enantiomer C-10c (62.00 mg, 84.38 μmol) in DCM (3.01 mL) was cooled to 0 °C in an ice / water bath and stirred. Then TFA (481.05 mg, 4.22 mmol, 325.03 μL) was added dropwise to the reaction solution. The resulting mixture was stirred for 2 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by C18 flash chromatography eluting with a concentration gradient of 0% to 50% MeCN in water (6 mmol / L NH 4 HCO 3 ) to give nitrooxy ester Example 10, enantiomer 1 (40 mg, 89% yield) as a white solid. LCMS: 3-minute chromatography (5 - 95% MeCN in water (6 mmol / L NH4 HCO 3 ) in 5 - 95% MeCN, Waters Acquity UPLC BEH C18 1.7um, 2.1×50mm, 40 °C) at t R = 1.63 min, MS(ESI) m / z = 535.3 [M+H] + . 1 H NMR: (500 MHz, DMSO-d 6 ) δ 12.85 (s, 1H), 10.81 (s, 1H), 7.95 (s, 2H), 7.57 (d, J = 8.8 Hz, 2H), 7.51 - 7.20 (m, 5H), 6.98 (t, J = 7.5 Hz, 1H), 5.06 (s, 2H), 4.48 (t, J = 6.2 Hz, 2H), 3.98 - 3.84 (m, 1H), 3.29 - 3.26 (m, 1H), 2.98 - 2.84 (m, 1H), 2.35 (t, J = 7.0 Hz, 2H), 1.75 - 1.47 (m, 4H), 1.42 - 1.27 (m, 2H). Example 11 (Enantiomer 1) Preparation of 4-(1-((3-(1H-Pyrazol-4-yl)-1H-indol-7-yl)amino)-3-amino-1-oxopropan-2-yl)benzyl 5-(nitrooxy)pentanoate

[0399]

Chemical Structure

[0400]

[0372] Step 1: A mixture of 5-bromopentanoic acid I-11a (1 g, 5.52 mmol, 1 equivalent) and AgNO 3 (1.9 g, 11.05 mmol, 2 equivalents) in MeCN (15 mL) was stirred at 70 °C for 24 hours under a nitrogen atmosphere. The reaction was photosensitive and light had to be avoided. The mixture was cooled to room temperature. The precipitated solid was collected by filtration and washed with dichloromethane (5 × 40 mL). After filtration, the filtrate was concentrated under reduced pressure to obtain 5-(nitrooxy)pentanoic acid I-11b (600 mg) as a colorless oil (67%).1 H NMR (DMSO-d 6 , 400 MHz) δ 1.58 - 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).

[0373] Step 2: To a mixture of 5-(nitrooxy)pentanoic acid I-11b (120 mg, 0.74 mmol, 5 equiv) in DCM (10 mL) was added EDCI (57 mg, 0.30 mmol, 2 equiv) and DMAP (2 mg, 0.016 mmol, 0.1 equiv) at 0 °C for 30 min under a nitrogen atmosphere, followed by the dropwise addition of alcohol C-1c (85 mg, 0.15 mmol, 1 equiv) at 0 °C. After warming the mixture to room temperature, the reaction solution was stirred for 16 h. The reaction was quenched with water (20 mL) at room temperature and extracted with CH 2 Cl 2 (3 × 20 mL). The combined organic layers were washed with saturated NaCl (3 × 20 mL) and dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC eluting with dichloromethane / methanol (30:1, v / v) to afford enantiomer C-11 (60 mg) as an orange solid (40%). LCMS: m / z (ESI), [M + H] + = 721.31.

[0401]

[0374] Step 3: A mixture of enantiomer C-11 (60 mg, 0.08 mmol, 1 equiv) and TFA (0.8 mL, 10.74 mmol) in DCM (3 mL) was stirred at 0 °C for 30 min under a nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was neutralized to pH 7 - 8 with saturated NaHCO 3 and extracted with CH 2 Cl 2 (3 × 10 mL). The combined organic layers were washed with saturated NaCl (3 × 10 mL) and dried over anhydrous Na 2 SO 4 . After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative HPLC using the following conditions (mobile phase: 0.1% NH 4 HCO 3and 0.1% NH in acetonitrile 4 It was composed of a solvent mixture of OH. A constant concentration gradient of 70% aqueous / 30% organic to 47% aqueous / 53% organic mobile phase was used over 8 minutes. The flow rate was constant at 60 mL / min. ) was purified to obtain Example 11, Enantiomer 1 (10.6 mg) as an orange solid (24%). LCMS: m / z (ESI), [M+H] + = 521.25 1 H NMR (DMSO-d 6 , 400 MHz) δ 1.60 - 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). Example 12 (Enantiomer 1) Preparation of 4-(1-((3-(1H-Pyrazol-4-yl)-1H-indol-7-yl)amino)-3-amino-1-oxopropan-2-yl)benzyl 4-(nitrooxy)butanoate

[0402]

Chemical Structure

[0403]

[0375] Step 1: To a mixture of 4-bromobutanoic acid C-12a (290 mg, 1.75 mmol, 5 equivalents) in DCM (30 mL) was added EDCI (133 mg, 0.70 mmol, 2 equivalents) and DMAP (4.2 mg, 0.04 mmol, 0.1 equivalent) at 0 °C for 30 minutes under a nitrogen atmosphere, followed by addition of alcohol C-1c (200 mg, 0.35 mmol, 1 equivalent). The reaction mixture was stirred at room temperature for 2 hours. The mixture was quenched with water (30 mL), and CH 2 Cl 2(3 × 30 mL) was extracted, washed with saturated NaCl (3 × 20 mL), and dehydrated with anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC eluting with dichloromethane / methanol (30 / 1, v / v) to give (1H-indol-3-yl)-1H-bromoester C-12b as a yellow solid (220 mg, 88% yield). LCMS: m / z (ESI), [M+H] + = 724.23.

[0404]

[0376] Step 2: A mixture of bromoester C-12b (120 mg, 0.17 mmol, 1 equiv) and AgNO 3 (56 mg, 0.34 mmol, 2 equiv) in MeCN (15 mL) was stirred at 70 °C for 2 h under a nitrogen atmosphere. The reaction was photosensitive and light had to be avoided. The mixture was cooled to room temperature. The resulting mixture was filtered and the filter cake was washed with CH 2 Cl 2 (3 × 50 mL) and concentrated under reduced pressure to give nitrooxyester C-12c as an orange solid (25 mg, 26% yield). LCMS: m / z (ESI), [M+H] + = 707.2.

[0405]

[0377] Step 3: A mixture of nitrooxyester C-12c (25 mg, 0.04 mmol, 1 equiv) and TFA (0.6 mL, 8 mmol) in DCM (3 mL) was stirred at 0 °C for 30 min under a nitrogen atmosphere. The mixture was neutralized to pH 7 - 8 with ammonia and concentrated under reduced pressure. The residue was further purified by preparative HPLC using the following conditions (The mobile phase consisted of a solvent mixture of 0.1% formic acid in water and acetonitrile. A constant concentration gradient from 83% aqueous / 17% organic to 70% aqueous / 30% organic mobile phase was used over 7 min. The flow rate was constant at 70 mL / min.) to give Example 12, enantiomer 1 as a white solid (2 mg, 9% yield). LCMS: m / z (ESI), [M+H] + = 507.35. 1 H NMR: (DMSO-d 6, 400 MHz) δ 1.92 - 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). Example 14 (R)-N-(3-(1H-Pyrazol-4-yl)-1H-indol-7-yl)-3-amino-2-phenylpropanamide Preparation

[0406] [Chemical Structure]

[0407]

[0378] Step 1: 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 PdCl 2 dppf (705.11 mg, 863.43 μmol) in dioxane (60.00 mL) and water (12.00 mL) were stirred at 90 °C for 16 h under nitrogen. The solvent was removed under reduced pressure. The residue was purified by silica chromatography column eluting with a concentration gradient of 10 to 50% ethyl acetate in petroleum ether. The pure fractions were evaporated to dryness to give amine I-14b (0.471 g, 748.56 μmol, yield 9%, purity 72%). LCMS: 3-minute chromatography (3 minutes - 5 - 95% MeCN in water (0.02% TFA), Waters Acquity UPLC BEH C18 1.7um, 2.1×50mm, 40 °C) with t R = 1.77 minutes, MS (ESI) m / z = 453.6 [M + H] +And amine I-14a (3.19 g, 6.88 mmol, yield 80%, purity 76%) was obtained as a brown solid. LCMS: 3-minute chromatography (3 minutes - 5 - 95% MeCN in water (0.02% TFA), Waters Acquity UPLC BEH C18 1.7 um, 2.1×50 mm, 40 °C), tR = 1.43 minutes, MS(ESI) m / z = 353.6 [M+H] + .

[0408]

[0379] Step 2: A stirred mixture of amine I-14b (85 mg, 135.09 μmol), (R)-3-((tert-butoxycarbonyl)amino)-2-phenylpropanoic acid (39.42 mg, 148.60 μmol) and DIPEA (26.19 mg, 202.64 μmol, 35.29 μL) in NMP (2.00 mL) was cooled and stirred at 0 °C. Then HATU (56.80 mg, 149.39 μmol) was added all at once at this temperature. The resulting mixture was stirred at room temperature for 17 hours. Then, when the reaction was detected by LCMS, the reaction was not complete. Further, (R)-3-((tert-butoxycarbonyl)amino)-2-phenylpropanoic acid (107.52 mg, 405.27 μmol) was added to the reaction solution. The reaction mixture was cooled and stirred at 0 °C. Then, further DIPEA (69.84 mg, 540.36 μmol, 94.12 μL) was added all at once at this temperature. The resulting mixture was stirred at room temperature for 31 hours. The solution was purified by a C18 flash chromatography column eluting with a concentration gradient of 0 to 80% MeCN in water (6 mmol / L NH 4 HCO 3 ). The pure fractions were lyophilized to obtain amide (R)-enantiomer C-14c (15 mg, yield 16%) as a white solid. LCMS: 3-minute chromatography (3 minutes - 5 - 95% MeCN in water (0.02% TFA), Waters Acquity UPLC BEH C18 1.7 um, 2.1×50 mm, 40 °C), t R = 2.18 minutes, MS(ESI) m / z = 700.5 [M+H] + .

[0409]

[0380] Step 3: A solution of (R)-enantiomer C-14c (15 mg, 21.43 μmol) in DCM (1.00 mL) was cooled to 0 °C in an ice / water bath and stirred. Then 4M HCl (72.76 mg, 2.00 mmol, 0.50 mL) was added dropwise to the reaction solution. 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 flash chromatography eluting with a concentration gradient of 0% to 50% MeCN in water (6 mmol / L NH 4 HCO 3 ). The pure fractions were lyophilized to give (R)-enantiomer C-14d (10 mg, 93% yield) as a white solid. LCMS: 3-minute chromatography (3 minutes - 5 to 95% MeCN in water (0.02% TFA), Waters Acquity UPLC BEH C18 1.7um, 2.1×50 mm, 40 °C) with t R = 1.59 minutes, MS (ESI) m / z = 500.2 [M+H] + .

[0410]

[0381] Step 4: A solution of (R)-enantiomer C-14d (10 mg, 20.02 μmol) was dissolved in 1M TBAF in THF (104.67 mg, 400.34 μmol, 0.40 mL). The reaction solution was sealed and heated at 80 °C for 16 hours. The reaction solution was evaporated under reduced pressure. The residue was purified by a C18-flash chromatograph column eluting with a concentration gradient of 0 to 30% CH 3 CN in water (0.02% TFA). The pure fractions were evaporated to dryness to give (R)-enantiomer Example 14 (4 mg, 8.73 μmol, 44% yield, TFA salt) as a brown solid. LCMS: 3-minute chromatography (3 minutes - 5 to 95% MeCN in water (0.02% TFA), Waters Acquity UPLC BEH C18 1.7um, 2.1×50 mm, 40 °C) with t R = 1.23 minutes, MS (ESI) m / z = 346.3 [M+H] + . 1 H NMR: (400 MHz, DMSO-d 6) δ 10.75 (s, 1H), 10.20 (s, 1H), 7.97 (d, J = 12.6 Hz, 5H), 7.66 - 7.56 (m, 2H), 7.48 (d, J = 7.4 Hz, 2H), 7.42 (t, J = 7.4 Hz, 2H), 7.35 (d, J = 7.5 Hz, 2H), 6.99 (t, J = 7.9 Hz, 1H), 4.21 (dd, J = 9.4, 4.6 Hz, 1H), 3.58 - 3.55 (m, 1H), 3.16 - 3.12 (m, 1H). Example 15 (S)-N-(3-(1H-Pyrazol-4-yl)-1H-indol-7-yl)-3-amino-2-phenylpropanamide Preparation

[0411]

Chemical Structure

[0412]

[0382] Step 1: Nitro-indole I-15a (219 mg, 641.93 μmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-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 PdCl 2 dppf (52.42 mg, 64.19 μmol) in dioxane (5.00 mL) and water (1 mL) were stirred at 90 °C for 16 h under nitrogen. The solvent was removed under reduced pressure. The residue was purified by silica column chromatography eluting with a concentration gradient of 0 to 10% methanol in DCM. The pure fractions were evaporated to dryness to give pyrazole I-15b (200 mg, 87% yield) as a brownish solid. LCMS: 3-minute chromatography (3 minutes - 5 - 95% MeCN in water (0.02% TFA), Waters Acquity UPLC BEH C18 1.7um, 2.1×50 mm, 40 °C) with t R = 2.01 minutes, MS (ESI) m / z = 359.3 [M + H]+ .

[0413]

[0383] 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 h. The reaction mixture was filtered through celite, washed with ethanol (10 mL × 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 obtain amine I-15c (160 mg, 87% yield) as a brownish solid. LCMS: 3-minute chromatography (3 minutes - 5 - 95% MeCN in water (0.02% TFA), Waters Acquity UPLC BEH C18 1.7um, 2.1×50 mm, 40 °C) with t R = 1.56 min, MS (ESI) m / z = 329.3 [M+H] + .

[0414]

[0384] Step 3: A stirred mixture of (S)-3-((tert-butoxycarbonyl)amino)-2-phenylpropanoic 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 at 0 °C. Then HATU (69.82 mg, 183.63 μmol) was added all at once at this temperature. The resulting mixture was stirred at 0 °C for 2 h. The solution was purified by C18 flash chromatography column eluting with a concentration gradient of 0 to 60% MeCN in water. The pure fractions were lyophilized to obtain amide (S)-enantiomer C-15 (38 mg, 43% yield) as a white solid. LCMS: 3-minute chromatography (3 minutes - 5 - 95% MeCN in water (0.02% TFA), Waters Acquity UPLC BEH C18 1.7um, 2.1×50 mm, 40 °C) with t R = 2.08 min, MS (ESI) m / z = 576.6 [M+H] + .

[0415]

[0385] Step 4: A solution of amide (S)-enantiomer C-15 (38 mg, 66.00 μmol) in DCM (2.00 mL) was cooled to 0 °C in an ice / water bath and stirred. Then 4M HCl (145.83 mg, 4.00 mmol, 1.00 mL) was added dropwise to the reaction solution. 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 flash chromatography eluting with a concentration gradient of 0% to 50% MeCN in water (6 mmol / L NH 4 HCO 3 ). The pure fractions were lyophilized to give (S)-enantiomer Example 15 (3.2 mg, 14% yield) as a white solid. LCMS: 3-minute chromatography (3 minutes - 5 to 95% MeCN in water (0.02% TFA), Waters Acquity UPLC BEH C18 1.7um, 2.1×50 mm, 40 °C) with t R = 1.17 minutes, MS (ESI) m / z = 346.2 [M+H] + . 1 1H NMR: (400 MHz, DMSO-d 6 ) δ 10.87 (s, 1H), 7.95 (s, 2H), 7.58 (d, J = 8.1 Hz, 2H), 7.43 (t, J = 8.8 Hz, 3H), 7.40 - 7.32 (m, 3H), 7.29 (t, J = 7.1 Hz, 1H), 6.98 (t, J = 7.7 Hz, 1H), 4.07 - 3.97 (m, 1H), 3.61 - 3.56 (m, 1H), 3.00 - 2.93 (m, 1H). Example 16 (racemate), Example 17 (enantiomer 1) and Example 18 (enantiomer 2) Preparation of N-(3-(1H-pyrazol-4-yl)-1H-indol-7-yl)-3-amino-2-(3-methoxyphenyl)propanamide

[0416]

Chemical formula

[0417]

[0386] Step 1: In a 20 mL vial, at room temperature, CH 3 Acid I-2 (237.58 mg, 0.80 mmol, 1.2 eq), TCFH (282.14 mg, 1.01 mmol, 1.5 eq), NMI (220.16 mg, 2.68 mmol, 4 eq) and amine I-49 (200 mg, 0.67 mmol, 1.00 eq) in CN (1.5 mL) were added. The resulting mixture was stirred at room temperature for 2 h under a nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was purified by preparative TLC using DCM / methanol (30:1, v / v) to obtain amide C-16a (150 mg, 38.87%) as a yellow solid. LCMS: m / z (ESI), [M+H] + =576.30.

[0418]

[0387] Step 2: The racemic amide C-16a was purified by preparative chiral HPLC using the following conditions (column: CHIRALPAK IG, 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; concentration gradient: 25% B in 23 min; wavelength: 220 / 254 nm; t R -1 = 14.66 min; t R -2 = 19.11 min; sample solvent: EtOH:DCM = 1:1, v / v; injection volume: 0.3 mL) to obtain enantiomer C-16b and enantiomer C-16c as white solids.

[0419]

[0388] Step 3: In a 50 mL round-bottom flask, at room temperature, enantiomer C-16c (50 mg, 0.087 mmol, 1 eq) in TFA (1 mL) and DCM (2 mL) were added. The resulting mixture was stirred at room temperature for 1.5 h. The mixture was basified to pH 9 with saturated NaHCO 3 (aqueous solution). The resulting mixture was extracted with CH 2 Cl 2 (3×10 mL). The combined organic layers were concentrated under reduced pressure. The crude product was purified by preparative HPLC using the following conditions (column: XBridge Prep OBD 30×150 mm, 5 μm; mobile phase A: water (10 mmol / L NH 4 HCO3 ) Mobile phase B: ACN; Flow rate: 60 mL / min; Concentration gradient: 10% B to 40% B in 7 minutes; Wavelength: 220 nm; t R = 6.25 minutes) for purification, and Example 18, enantiomer 2 (14.0 mg, 42.93%) was obtained as a white solid. LCMS: m / z (ESI), [M+H] + = 376.15. 1 H NMR (CD...

Claims

1. Equation I 【Chemistry 1】 [In the formula, R １ and R ２ Each of these is independently selected from the group consisting of hydrogen, hydroxyl, halogen, cyano, amino, alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, and heteroalkynyl, and each of the alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, and heteroalkynyl is optionally substituted with one or more groups independently selected from hydroxyl, halogen, cyano, or amino. Each of X, W, Z, and U is independently N or C(R ３ ) and R ３ The group is selected from the group consisting of hydrogen, hydroxyl, halogen, cyano, amino, alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, and heteroalkynyl, and each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, and heteroalkynyl is optionally substituted with one or more groups independently selected from hydroxyl, halogen, cyano, or amino. E is -N(R ４ )-C(=O)-, -C(=O)N(R ４ )-, -N(R ４ )SO ２ -, or -SO ２ N(R ４ )-, and R ４ This is selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, and heteroalkynyl. Y is -Y １ -Y ２ - (Y ３ ) ｎ And in the formula, Y １ It does not exist, or -C(R ５ ) ２ - and Each R ５ is hydrogen, -N(R ａ ) ２ Independently selected from the group consisting of 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 one or more R ６ And it is replaced by optional selection. Each R ６ These include hydroxy, halogen, cyano, amino, alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, and -alkyl-N(R) ａ ) ２ Independently selected from the group consisting of, Each R ａ It is independently selected from hydrogen or alkyl, Y ２ is, -(CH ２ ) ｐ -Cycloalkyl-*, -(CH ２ ) ｐ -Heterocyclyl-*, -(CH ２ ) ｐ -aryl-*, or -(CH ２ ) ｐ -Heteroaryl-* is selected, each of which may be optionally substituted with one or more groups independently selected from hydroxy, halogen, cyano, amino, alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, or heteroalkynyl, where p is 0, 1, or 2, Y ２ The *end is Y ３ It is connected, Y ３ It does not exist, or -Y ３１ -Y ３２ -Y ３３ And in the formula, Y ３１ It is either absent or selected from alkyl, alkenyl, or alkynyl. Y ３２ It does not exist, or -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 ) - Selected from #, each R ｂ Y is independently selected from hydrogen or alkyl. ３２ The end of # is Y ３３ It is connected, Y ３３ is hydrogen, hydroxyl, cyano, halogen, -N(R) ｃ ) ２ Selected from the group consisting of 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 one or more R ７ It is replaced by arbitrary selection, and each R ７ R is independently selected from halogen, hydroxy, amino, cyano, nitrooxy, alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, or haloalkyl, and each R ｃ It is independently selected from hydrogen or alkyl, Furthermore n is an integer between 1 and 5. Compounds of or pharmaceutically acceptable salts thereof.

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

3. (i) R １ and R ２ Both are hydrogen; (ii) Both R1 and R2 are alkyl, and optionally R1 is methyl and R2 is methyl; (iii) R1 is hydrogen, R2 is halogen, and optionally R1 is hydrogen and R2 is fluoro; or (iv) R1 is hydrogen, R2 is alkyl, and optionally R1 is hydrogen and R2 is methyl. The compound according to claim 2 or a pharmaceutically acceptable salt thereof.

4. (i) X, W, U and Z are C(R ３ ) or (ii) X and U are C(R3), and W and Z are N, The compound according to claim 1 or a pharmaceutically acceptable salt thereof.

5. R ３ The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound is hydrogen.

6. E is -N(R ４ )C(=O)- or -C(=O)N(R ４ ) - wherein R4 is optionally hydrogen, the compound according to claim 1 or a pharmaceutically acceptable salt thereof.

7. (i) one R ５ R is hydrogen, and the other R ５ However, -N(R ａ ) ２ Selected from alkyl, cycloalkyl, heterocyclyl, or -alkyl-heterocyclyl, the alkyl, cycloalkyl, heterocyclyl, and -alkyl-heterocyclyl are one or more R ６ Replaced by optional selection; or (ii) One R5 is alkyl, and the other R5 is selected from -N(Ra)2, alkyl, cycloalkyl, or heterocyclyl, and the alkyl, cycloalkyl, and heterocyclyl are optionally substituted with one or more R6. The compound according to claim 1 or a pharmaceutically acceptable salt thereof.

8. R ６ However, amino, alkyl, or -alkyl-N(R) ａ ) ２ A compound according to claim 7 or a pharmaceutically acceptable salt thereof, selected from the above.

9. Y １ but, 【Chemistry 2】 A compound according to claim 7 or a pharmaceutically acceptable salt thereof, selected from the group consisting of the following.

10. (i) Y ２ ga- (CH ２ ) ｐ - (C ３～１０ It is a cycloalkyl)-*, where p is 0 or 1, and optionally Y2 is cyclopentyl or cyclohexyl; (ii) Y2 is -(CH2)p-(5- to 12-membered heterocyclyl)-*, where p is 0 or 1, and optionally Y2 is tetrahydrofuranil, pyrrolidinil, or tetrahydropyranil; (iii) Y2 is -(CH2)p-(C5-12aryl)-*, where p is 0 or 1, and optionally Y2 is phenyl or -CH2-phenyl-*; or (iv) Y2 is -(CH2)p-(5- to 12-membered heteroaryl)-*, where p is 0 or 1, and optionally Y2 is pyridinyl, tetrahydroisoquinolinyl, -CH2-imidazolyl-*, or -CH2-indolyl-*. The compound according to claim 1 or a pharmaceutically acceptable salt thereof.

11. Y ３１ A compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein the compound is not present.

12. Y ３２ However, they do not exist, or -O-#, -C(=O)O-#, -P(=O)(R ｂ )-#,-N(R ｂ )C(=O)-#, or -C(=O)N(R ｂ A compound according to claim 11 or a pharmaceutically acceptable salt thereof, selected from )-#.

13. Y ３３ However, hydrogen, hydroxyl, cyano, halogen, -N(R) ｃ ) ２ Selected from alkyl or heteroalkyl, the alkyl and heteroalkyl are one or more R ７ A compound according to claim 11 or a pharmaceutically acceptable salt thereof, which is optionally substituted by [a specific compound].

14. Y ３３ is hydrogen, hydroxyl, cyano, halogen, -NH ２ , methyl, -CH ２ CH ２ OCH ３ and is selected from the group consisting of, the compound according to claim 13 or a pharmaceutically acceptable salt thereof.

15. Y ３１ The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound is alkyl.

16. Y ３２ However, they do not exist, or -O-#, -OC(=O)-#, -OC(=O)N(R ｂ )-#, or-N(R ｂ A compound according to claim 15 or a pharmaceutically acceptable salt thereof, selected from C(=O)-#.

17. Y ３３ is selected from -N(R ｃ ), alkyl, or aryl, and the alkyl and aryl are optionally substituted with one or more R ２ s, the compound according to claim 15 or a pharmaceutically acceptable salt thereof.​​

18. Each R ７ The compound according to claim 17 or a pharmaceutically acceptable salt thereof, wherein the compound is independently selected from halogen, hydroxy, amino, cyano, nitrooxy, or alkyl.

19. Y ３３ However, -NH ２ , -N(CH ３ ) ２ A compound according to claim 18 or a pharmaceutically acceptable salt thereof, selected from methyl, dimethylphenyl, or nitrooxypentyl.

20. A compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein n is 1 or 2. 【Request Item 21】 【Chemistry 3-1】 【Chemistry 3-2】 【Chemistry 3-3】 [Chemistry 3-4] [Transformation 3-5] [Chemistry 3-6] 【Chemistry 3-7】 【Transformation 3-8】 【Chemistry 3-9】 【Chemistry 3-10】 【Chemistry 3-11】 【Chemistry 3-12】 【Chemistry 3-13】 【Chemistry 3-14】 A compound according to claim 1 or a pharmaceutically acceptable salt thereof, selected from the group consisting of the following.

22. A pharmaceutical composition comprising a compound according to any one of claims 1 to 21 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

23. The pharmaceutical composition according to claim 22, formulated as eye drops.

24. A pharmaceutical composition for inhibiting ROCK activity in subjects requiring inhibition of ROCK activity, comprising an effective amount of a compound according to any one of claims 1 to 21 or a pharmaceutically acceptable salt thereof.

25. A pharmaceutical composition for treating rock-related disorders in subjects requiring treatment of rock-related disorders, comprising an effective amount of a compound according to any one of claims 1 to 21 or a pharmaceutically acceptable salt thereof.

26. The pharmaceutical composition according to claim 24 or 25, wherein the ROCK-related disorder is an eye disease including glaucoma and retinal diseases such as exudative AMD, atrophic AMD and DME; a bone disorder including osteoporosis; cerebral vasospasm, coronary artery spasm; hypertension, pulmonary hypertension, sudden death syndrome, angina; myocardial infarction, restenosis, stroke; hypertensive vascular disease; heart failure; vascular diseases including allograft vascular disease and venous graft lesions; lung diseases including chronic obstructive pulmonary disease (COPD) and asthma; spinal cord injury; Alzheimer's disease; multiple sclerosis; neurological disorders including depression, attention deficit hyperactivity disorder and neuropathic pain; neovascularization and cancer; obesity; and erectile dysfunction, optionally wherein the ROCK-related disorder is glaucoma.

27. ​​A pharmaceutical composition comprising a compound according to any one of claims 1 to 21 or a pharmaceutically acceptable salt thereof, which can be combined with one or more additional therapeutic agents, wherein the one or more additional therapeutic agents are optionally selected from beta-blockers, alpha-agonists, carbonic anhydrase inhibitors, prostaglandin-like compounds, miotics or cholinergics, or epinephrine compounds.

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