HK40090358A - Atr inhibitors and uses thereof - Google Patents

Description

ATR inhibitors and their uses

Technical Field

This disclosure generally relates to novel compounds that can be used as ATR inhibitors, pharmaceutical compositions comprising these compounds, and methods of treatment by administering these compounds or the pharmaceutical compositions.

Background Technology

ATR (also known as FRAP-associated protein 1; FRP1, MEC1, SCKL, SECKL1) protein kinases are members of the PI3-kinase-like kinase (PIKK) protein family, involved in the repair, maintenance, and stability of the genome. They are crucial for the viability of replicating cells and are activated during S phase to regulate the initiation of replication origins and repair damaged replication forks. Therefore, ATR inhibitors have the potential to be an effective approach for cancer treatment.

Although progress has been made in ATR inhibitors, there remains a strong need in the field to develop improved drugs with inhibitory activity against ATR.

Summary of the Invention

This disclosure provides compounds capable of inhibiting ATR protein kinases, including their stereoisomers, pharmaceutically acceptable salts, tautomers, and prodrugs. Methods for using such compounds to treat various diseases or conditions, such as cancer, are also provided.

On the one hand, this disclosure provides a compound having formula (I):

Or its pharmaceutically acceptable salt.

in

Ring A is absent, or is a 3- to 6-membered cycloalkyl, 5- to 6-membered heterocyclic or 5- to 6-membered heteroaryl;

V is a direct bond, a carbonyl group, or an alkyl group optionally substituted with one or more R ^c groups;

W and L are each independently a direct bond, -O-, -S-, or -N( ^Ra )-;

^R1 is an alkyl group, a cyano group, -S(O) _{2CH3 ,} or -S(O)(NH) _CH3 ;

^R2 is hydrogen, halogen, or an alkyl group optionally substituted with one or more ^Rb ;

Ring B is

R ⁵ is selected from the group consisting of: hydrogen, halogen, hydroxyl, cyano, alkyl, alkenyl, ynyl, heteroalkyl, heteroalkenyl, heteroynyl, and haloalkyl;

^Ra is hydrogen or alkyl;

^Rb is a hydroxyl group or a halogen;

^Rc is a hydroxyl group, halogen, or alkyl group;

n can be 0, 1, 2, or 3.

In some embodiments, this disclosure provides a compound having formula (II) or formula (III):

In some embodiments, this disclosure provides a compound having the formula selected from the group consisting of:

as well as

in

U is either O or NH;

V is a direct bond, a carbonyl group, or an alkyl group optionally substituted with one or more R ^c groups;

W and L are each independently -N( ^Ra )-;

^R1 is an alkyl group;

^R2 is hydrogen, halogen, or an alkyl group substituted with one or more ^Rb ;

R ⁵ is hydrogen or alkyl;

^Ra is hydrogen or alkyl;

^Rb is a hydroxyl group or a halogen; and

^Rc is a hydroxyl group, a halogen, or an alkyl group.

In some embodiments, this disclosure provides a compound having formula (V):

Or its pharmaceutically acceptable salt.

in

Ring A is absent, or is a 3- to 6-membered cycloalkyl, 5- to 6-membered heterocyclic, or 5- to 6-membered heteroaryl; Q is a direct bond or an alkyl group optionally substituted with one or more R ^d ;

L is -O-, -S-, or -N( ^Ra )-;

Ring B is

^Ra is hydrogen or alkyl;

R ^_d is a hydroxyl group, a halogen, or an alkyl group;

^R1 is selected from the group consisting of: cyano, hydroxyl, halogen, -H, -S(O) _2CH3 and -S(O)(NH) _{CH3 ;}

R ⁵ is selected from the group consisting of: hydrogen, halogen, hydroxyl, cyano, alkyl, alkenyl, ynyl, heteroalkyl, heteroalkenyl, heteroynyl, and haloalkyl;

n can be 0, 1, 2, or 3.

On the other hand, this disclosure provides a pharmaceutical composition comprising a compound of the disclosure or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

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

In another aspect, this disclosure provides the use of a compound of the present disclosure or a pharmaceutically acceptable salt thereof or a pharmaceutical composition thereof in the preparation of a medicament for the prevention or treatment of cancer.

In another aspect, this disclosure provides compounds of the present disclosure or pharmaceutically acceptable salts thereof or pharmaceutical compositions thereof for the treatment of cancer.

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

Detailed Implementation

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

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

definition

The definitions of specific functional groups and chemical terms are described in more detail below. For the purposes of this disclosure, chemical elements are identified according to the Periodic Table of the Elements, CAS edition, Handbook of Chemistry and Physics, 75th edition, inner cover, and specific functional groups are generally defined as described herein. In addition, the general principles of organic chemistry, as well as specific functional components and reactivity, are described in the following literature: *Organic Chemistry*, Thomas Sorrell, 2nd ed., University Science Books, Sausalito, 2006; Smith and March, *March's Advanced Organic Chemistry*, 6th ed., John Wiley & Sons, Inc., New York, 2007; Lar ock, *Comprehensive Organic Transformations*, 3rd ed., VCH Publishers, Inc., New York, 2018; Carruthers, *Some Modern Methods of Organic Synthesis*, 4th ed., Cambridge University Press, Cambridge, 2004; The entire contents of each of the above references are incorporated herein by reference.

Linking substituents are described throughout this disclosure. Where the structure explicitly requires a linking group, the Markush variable listed with respect to said group should be understood as the linking group. For example, if the structure requires a linking group and the Markush group definition of said variable lists "alkyl", then "alkyl" should be understood to mean a linked alkylene group.

When the bond connecting the substituent to the substituent crosses with the bond between two atoms in the linking ring, such a substituent may bond to any atom in the ring. When a substituent is listed but not specified through which atom it bonds to the remainder of the compound in the given formula, the substituent may bond to any atom in the formula. Combinations of substituents and/or variables are permitted, but only if such combinations produce a stable compound.

When any variable (e.g., ^Ri ) appears more than once in any component or formula of a compound, its definition for each occurrence is independent of its definition for each subsequent occurrence. Thus, for example, if a substituent group is substituted by 0 to 2 ^Ri moieties, the substituent group may optionally be substituted by at most two ^Ri moieties, and ^Ri is independently selected from the definition of ^Ri each time it appears. Furthermore, combinations of substituents and/or variables are permitted, but only if such combinations produce a stable compound.

As used herein, the term "C_{_ij}" indicates a range of carbon atoms, where i and j are integers, and the range includes the endpoints (i.e., i and j) and every integer point in between, where j is greater than i. For example, C _{_1-6} indicates a range of one to six carbon atoms, including one carbon atom, two carbon atoms, three carbon atoms, four carbon atoms, five carbon atoms, and six carbon atoms. In some embodiments, the term "C _{_1-12} " indicates 1 to 12 carbon atoms, particularly 1 to 10 carbon atoms, particularly 1 to 8 carbon atoms, particularly 1 to 6 carbon atoms, particularly 1 to 5 carbon atoms, particularly 1 to 4 carbon atoms, particularly 1 to 3 carbon atoms, or particularly 1 to 2 carbon atoms.

As used herein, the term "alkyl," whether used as part of another term or independently, refers to a saturated straight-chain or branched hydrocarbon group that may optionally be substituted independently by one or more substituents described below. The term "C_{_ij}alkyl" refers to an alkyl group having i to j carbon atoms. In some embodiments, the alkyl group contains 1 to 10 carbon atoms. In some embodiments, the alkyl group contains 1 to 9 carbon atoms. In some embodiments, the alkyl group contains 1 to 8 carbon atoms, 1 to 7 carbon atoms, 1 to 6 carbon atoms, 1 to 5 carbon atoms, 1 to 4 carbon atoms, 1 to 3 carbon atoms, or 1 to 2 carbon atoms. Examples of "C _{_1-10}alkyl" include, but are not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, and decyl. Examples of "C _1-6 alkyl" include methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 2-pentyl, 3-pentyl, 2-methyl-2-butyl, 3-methyl-2-butyl, 3-methyl-1-butyl, 2-methyl-1-butyl, 1-hexyl, 2-hexyl, 3-hexyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 3-methyl-3-pentyl, 2-methyl-3-pentyl, 2,3-dimethyl-2-butyl, 3,3-dimethyl-2-butyl, etc.

As used herein, the term "alkenyl," whether used as part of another term or independently, refers to a straight-chain or branched hydrocarbon group having at least one carbon-carbon double bond that may optionally and independently be substituted by one or more substituents described herein, and includes groups 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, vinyl (ethylenyl or vinyl), allyl (allyl), butenyl, pentenyl, 1-methyl-2-buten-1-yl, 5-hexenyl, etc.

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

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 system in which all ring atoms are carbon, and the system contains at least three cyclic carbon atoms. In some embodiments, the cycloalkyl group may contain 3 to 12 cyclic carbon atoms, 3 to 10 cyclic carbon atoms, 3 to 9 cyclic carbon atoms, 3 to 8 cyclic carbon atoms, 3 to 7 cyclic carbon atoms, 3 to 6 cyclic carbon atoms, 3 to 5 cyclic carbon atoms, 4 to 12 cyclic carbon atoms, 4 to 10 cyclic carbon atoms, 4 to 9 cyclic carbon atoms, 4 to 8 cyclic carbon atoms, 4 to 7 cyclic carbon atoms, 4 to 6 cyclic carbon atoms, or 4 to 5 cyclic carbon atoms. The cycloalkyl group may be saturated or partially unsaturated. The cycloalkyl group may be substituted. In some embodiments, the cycloalkyl group may be a saturated cyclic alkyl group. In some embodiments, the cycloalkyl group can be a partially unsaturated cycloalkyl group containing at least one double or triple bond in its ring system. In some embodiments, the cycloalkyl group can be monocyclic or polycyclic. Examples of monocyclic cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopent-1-enyl, 1-cyclopent-2-enyl, 1-cyclopent-3-enyl, cyclohexyl, 1-cyclohex-1-enyl, 1-cyclohex-2-enyl, 1-cyclohex-3-enyl, cyclohexadienyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, cycloundecyl, and cyclododecyl. Examples of polycyclic cycloalkyl groups include, but are not limited to, adamantyl, norbornel, fluorenyl, spiropeptadienyl, spiro[3,6]decyl, bicyclo[1,1,1]pentenyl, bicyclo[2,2,1]heptenyl, etc.

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

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).

As used herein, the term "haloalkyl" refers to an alkyl group substituted with one or more halogens as defined above. Examples of haloalkyl groups include, but are not limited to, trifluoromethyl, difluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 1,2-difluoroethyl, 3-bromo-2-fluoropropyl, 1,2-dibromoethyl, etc.

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

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

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

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

As used in this article, the term "hydroxyl group" refers to -OH.

As used herein, the term "partially unsaturated" refers to a group comprising at least one double or triple bond. The term "partially unsaturated" is intended to cover rings having multiple unsaturated sites, but is not intended to include aromatic (i.e., fully unsaturated) moieties.

As used herein, the term “substituted,” whether or not preceded by the term “optionally,” means that one or more hydrogen atoms of the specified moiety are replaced by suitable substituents. It should be understood that “substituted” or “replaced by” includes the implicit precondition that such substitution is consistent with the permissible valence of the substituted atom and that the substitution yields a stable or chemically viable compound, e.g., a compound that does not spontaneously undergo transformations such as rearrangement, cyclization, or elimination. Unless otherwise stated, an “optionally substituted” group may have suitable substituents at each substituted position of the group, and at each position, the substituents may be the same or different when more than one position in any given structure can be replaced by more than one substituent selected from the specified group. Those skilled in the art will understand that the substituent itself can be substituted, if appropriate. Unless specifically stated as “unsubstituted,” references to chemical parts herein should be understood to include substituted variants. For example, a reference to an “aryl” group or moiety implicitly includes both substituted and unsubstituted variants.

compound

This disclosure provides a novel (I) compound and its pharmaceutically acceptable salt, a synthetic method for preparing said compound, pharmaceutical compositions containing the same, and various uses of the disclosed compound.

On the one hand, this disclosure provides a compound having formula (I):

Or its pharmaceutically acceptable salt.

in

Ring A is absent, or is a 3- to 6-membered cycloalkyl, 5- to 6-membered heterocyclic or 5- to 6-membered heteroaryl;

V is a direct bond, a carbonyl group, or an alkyl group optionally substituted with one or more R ^c groups;

W and L are each independently a direct bond, -O-, -S-, or -N( ^Ra )-;

^R1 is an alkyl group, a cyano group, -S(O) _{2CH3 ,} or -S(O)(NH) _CH3 ;

^R2 is hydrogen, halogen, or an alkyl group optionally substituted with one or more ^Rb ;

Ring B is

R ⁵ is selected from the group consisting of: hydrogen, halogen, hydroxyl, cyano, alkyl, alkenyl, ynyl, heteroalkyl, heteroalkenyl, heteroynyl, and haloalkyl;

^Ra is hydrogen or alkyl;

^Rb is a hydroxyl group or a halogen;

^Rc is a hydroxyl group, halogen, or alkyl group;

n can be 0, 1, 2, or 3.

In some implementations, V is a direct key.

In some implementations, V is a carbonyl group.

In some embodiments, V is an alkyl group optionally substituted with one or more ^Rc . In some embodiments, V is a _C1-6 alkyl, _C1-5 alkyl, _C1-4 alkyl, or _C1-3 alkyl.

In some implementations, ring A is absent.

In some embodiments, ring A is a 3- to 6-membered cycloalkyl group.

In some embodiments, ring A is cyclopropyl. In some embodiments, ring A is...

In some embodiments, ring A is cyclopentyl. In some embodiments, ring A is...

In some embodiments, ring A is cyclohexyl. In some embodiments, ring A is...

In some implementations, ring A is a 5- to 6-membered heterocyclic group.

In some embodiments, ring A is a 5-membered heterocyclic group containing at least one nitrogen atom. In some embodiments, ring A is a 5-membered heterocyclic group containing at least two nitrogen atoms. In some embodiments, ring A is a 5-membered heterocyclic group containing two nitrogen atoms.

In some embodiments, ring A is a pyrazolyl group.

In some implementations, ring A is a 6-membered heterocyclic group.

In some embodiments, ring A is a tetrahydropyranyl group.

In some embodiments, ring A is a 5- to 6-membered heteroaryl group.

In some embodiments, ring A is a 5- to 6-membered heteroaryl group containing at least one nitrogen atom.

In some embodiments, ring A is a 5-membered heteroaryl group containing at least one nitrogen atom. In some embodiments, ring A is a 5-membered heteroaryl group containing at least one nitrogen atom and an additional heteroatom selected from O, N, or S. In some embodiments, ring A is a thiazolyl, triazolyl, or isoxazolyl group.

In some embodiments, ring A is a 6-membered heteroaryl group containing at least one nitrogen atom. In some embodiments, ring A is a 6-membered heteroaryl group containing at least one nitrogen atom and an additional heteroatom selected from O, N, or S. In some embodiments, ring A is a pyridyl group.

In some implementations, W is a direct key.

In some implementations, W is -N( ^Ra )-.

In some implementations, W is -N( ^Ra )-, and ^Ra is hydrogen.

In some embodiments, W is -N( ^Ra )- and ^Ra is an alkyl group. In some embodiments, W is -N( ^Ra )- and Ra is a _C1-3 alkyl group. In some embodiments, W is -N( ^Ra )- and ^Ra is a methyl ^group .

In some embodiments, ring A is a 3- to 6-membered cycloalkyl, a 5- to 6-membered heterocyclic or a 5- to 6-membered heteroaryl, and W is a direct bond.

In some implementations, ring A does not exist, and W is -N( ^Ra )-.

In some implementations, ring A is absent, W is -N( ^Ra )-, and ^Ra is hydrogen.

In some embodiments, ring A is absent, W is -N( ^Ra )-, and ^Ra is an alkyl group. In some embodiments, ring A is absent, W is -N( ^Ra )-, and ^Ra is a _C1-3 alkyl group. In some embodiments, ring A is absent, W is -N( ^Ra )-, and ^Ra is a methyl group.

In some implementations, ring A does not exist, and W is a direct bond.

In some embodiments, ^R1 is an alkyl group.

In some embodiments, ^R1 is a _C1-3 alkyl group.

In some implementations, ^R1 is a cyano group.

In some embodiments, ^R1 is a hydroxyl group.

In some implementations, ^R1 is -S ₍ O) _2CH3 .

In some implementations, ^R1 is –S(O)(NH) _CH3 .

In some embodiments, ring A is absent, and ^R1 is _a cyano group or -S(O) _2CH3 .

In some embodiments, ring A is a 3- to 6-membered cycloalkyl, _{a 5- to 6-membered heterocyclic or a 5- to 6-membered heteroaryl, and R1 is an alkyl, hydroxyl, -S(O)2CH3} ^or _-S (O)(NH) _CH3 .

In some embodiments, ring A is cyclopropyl, cyclohexyl, tetrahydropyranyl, thiazolyl, pyridinyl _{, or isoxazolyl, and R1 is -S(O)2CH3} ^or _-S (O)(NH) _CH3 .

In some embodiments, ring A is cyclopropyl, _{and R1 is -S(O)₂CH₃} ^or _{-S ( O} ⁾ (NH) _{CH₃ .}

In some embodiments, ring A is cyclopropyl, ^R1 _{is -S(O)2CH3 or} -S ₍ O)(NH) _CH3 , and _n is ¹ _.

In some embodiments, ring A is cyclopropyl, W is a direct bond, ^R1 is ^-S _{(O)2CH3 or -S} (O)(NH) _{CH3 ,} and _n is 1.

In some implementations, for

In some embodiments, ring A is cyclopentyl and ^R1 is cyano. In some embodiments, ring A is cyclopentyl and ^R1 is cyano. In some embodiments, ring A is cyclopentyl, ^R1 is cyano, and n is 1.

In some embodiments, ring A is cyclopentyl, ^W is a direct bond, ^R1 is a cyano group, and n is 1.

In some implementations, for

In some embodiments, ring A is cyclohexyl and ^R1 is cyano. In some embodiments, ring A is cyclohexyl and ^R1 is cyano. In some embodiments, ring A is cyclohexyl, ^R1 is cyano, and n is 1.

In some embodiments, ring A is cyclopentyl, ^W is a direct bond, ^R1 is a cyano group, and n is 1.

In some implementations, for

In some embodiments, ring A is a 5-membered heterocyclic group, and ^R1 is an alkyl group.

In some embodiments, ring A is pyrazolyl, isoxazolyl, or triazolyl, and ^R1 is a _C1-3 alkyl group.

In some embodiments, ring A is pyrazolyl, isoxazolyl, or triazolyl, and ^R1 is methyl. In some embodiments, ring A is pyrazolyl, isoxazolyl, or triazolyl, ^R1 is methyl, and n is 2.

In some embodiments, ring A is pyrazolyl, isoxazolyl, or triazolyl, W is a direct bond, and ^R1 is a methyl group. In some embodiments, ring A is pyrazolyl, isoxazolyl, or triazolyl, W is a direct bond, ^R1 is a methyl group, and n is 2.

In some implementations, for

In some embodiments, ring A is _a 5- to 6-membered heteroaryl group, and ^R1 is -S(O) _2CH3 .

In some embodiments, ring A is thiazolyl or pyridinyl, and ^R1 is -S(O) _2CH3 . In some embodiments, ring A is thiazolyl or pyridinyl, ^R1 is -S(O _{)2CH3 ,} and n is ₁ .

In some embodiments, ring A is thiazolyl or pyridinyl, W is a direct bond, and ^R1 is -S(O) _2CH3 . In some embodiments, ring A is thiazolyl or pyridinyl, W is a direct bond, ^R1 is -S(O) _2CH3 , and _n is ₁ .

In some implementations, for

In some implementations, L stands for key.

In some implementations, L is -O-.

In some implementations, L stands for -S-.

In some implementations, L is -N( ^Ra )-.

In some implementations, ^Ra is hydrogen.

In some embodiments, ^Ra is a _C1-3 alkyl group.

In some implementations, ring B is

In some implementations, L is -O-, -S-, or -N( ^Ra )-, and ring B is

In some implementations, L is -O-, and ring B is

In some implementations, L is -S-, and ring B is

In some embodiments, L is -N( ^Ra )-, ^Ra is hydrogen, and ring B is...

In some embodiments, L is -N( ^Ra )-, ^Ra is hydrogen, and ring B is...

In some implementations, ^R2 is hydrogen.

In some embodiments, ^R2 is a halogen. In some embodiments, ^R2 is fluorine, chlorine, or bromine. In some embodiments, ^R2 is fluorine.

In some embodiments, ^R2 is an alkyl group substituted with one or more ^Rb . In some embodiments, ^R2 is a _C1-3 alkyl group substituted with one or more ^Rb .

In some embodiments, ^R2 is an alkyl group substituted with one or more ^Rb , and ^Rb is a hydroxyl group or fluorine. In some embodiments, ^R2 is a _C1-3 alkyl group substituted with one or more ^Rb , and ^Rb is a hydroxyl group or fluorine.

In some implementations, ^R2 is _-CH2OH or _-CH2F .

In some embodiments, this disclosure provides a compound having formula (II) or formula (III):

V, W, L, ring A, ring B, ^R1 and ^R2 are as defined above.

In some embodiments, this disclosure provides a compound having a formula selected from the group consisting of:

as well as

Or its pharmaceutically acceptable salt, wherein

U is either O or NH;

V is a direct bond, a carbonyl group, or an alkyl group optionally substituted with one or more R ^c groups;

W and L are each independently -O-, -S-, or -N( ^Ra )-;

^R1 is an alkyl group;

^R2 is hydrogen, halogen, or an alkyl group substituted with one or more ^Rb ;

^R3 is a halogen;

R ⁵ is hydrogen or alkyl;

^Ra is hydrogen or alkyl;

^Rb is a hydroxyl group or a halogen; and

^Rc is a hydroxyl group, a halogen, or an alkyl group.

In some embodiments, this disclosure provides a compound having a formula selected from the group consisting of:

Or its pharmaceutically acceptable salt.

In another aspect, this disclosure provides a compound having formula (V):

Or its pharmaceutically acceptable salt.

in

Ring A is absent, or is a 3- to 6-membered cycloalkyl, 5- to 6-membered heterocyclic or 5- to 6-membered heteroaryl;

Q is an alkyl group that is directly bonded or optionally substituted with one or more R ^d ;

L is -O-, -S-, or -N( ^Ra )-;

Ring B is

R ^_d is hydrogen or alkyl;

R ^_d is a hydroxyl group, a halogen, or an alkyl group;

^R1 is selected from the group consisting of: cyano, hydroxyl, halogen, -S( _O ) _2CH3 and -S(O)(NH) _CH3 ;

R ⁵ is selected from the group consisting of: hydrogen, halogen, hydroxyl, cyano, alkyl, alkenyl, ynyl, heteroalkyl, heteroalkenyl, heteroynyl, and haloalkyl;

n can be 0, 1, 2, or 3.

In some implementations, Q is a direct key.

In some embodiments, Q is an alkyl group. In some embodiments, Q is a _C1-6 alkyl, _C1-5 alkyl, _C1-4 alkyl, or _C1-3 alkyl.

In some embodiments, ring A is a 3- to 6-membered cycloalkyl group. In some embodiments, ring A is a cyclopropyl group. In some embodiments, ring A is...

In some implementations, ring A is absent.

In some embodiments, ring A is a 5- or 6-membered heterocyclic group. In some embodiments, ring A is a tetrahydropyranyl group.

In some implementations, ring A is

In some embodiments, Q is an alkyl group, and cyclic A is absent.

In some embodiments, Q is a direct bond, and ring A is a 3- to 6-membered cycloalkyl group or a 5- to 6-membered heterocyclic group.

In some implementations, ^R1 is -S _{(O)2CH3 or} -S(O)(NH) _CH3 .

In some embodiments, ^R1 is a cyano group, a hydroxyl group, or a halogen.

In some embodiments, ring A is absent, or is _{a 3- to 6-membered cycloalkyl or a 5- to 6-membered heterocyclic group, and R1 is -S(O)2CH3} ^or _-S (O)(NH) _CH3 .

In some embodiments, ring A is absent or is a 3- to 6-membered cycloalkyl group, and ^R1 is a cyano, hydroxyl, or halogen.

In some implementations, L is -O-.

In some implementations, L stands for -S-.

In some implementations, L is -N( ^Ra )-, and ^Ra is hydrogen.

In some implementations, ring B is

In some embodiments, ^R5 is hydrogen or alkyl.

In some embodiments, this disclosure provides a compound having a formula selected from the group consisting of:

Or its pharmaceutically acceptable salt.

Table 1 below illustrates exemplary compounds of this disclosure.

Table 1

The compounds described herein are based on general formulas and specific compounds. Furthermore, the compounds disclosed herein may exist in a variety of different forms or derivatives, including but not limited to prodrugs, soft drugs, active metabolic derivatives (active metabolites), and their pharmaceutically acceptable salts, all of which are within the scope of this disclosure.

As used herein, the term "prodrug" refers to a compound or a pharmaceutically acceptable salt thereof that, when metabolized under physiological conditions or converted via a solvolysis reaction, yields the desired active compound. Prodrugs include, but are not limited to, esters, amides, carbamates, carbonates, acylureas, solvates, or hydrates of the active compound. Typically, prodrugs are inactive or less active than the active compound, but may provide one or more advantageous disposal, administration, and/or metabolic properties. For example, some prodrugs are esters of the active compound; during metabolism, the ester group is cleaved to produce the active drug. Furthermore, some prodrugs are compounds that are enzymatically activated to produce the active compound or that produce the active compound after further chemical reactions. Prodrugs can develop from their prodrug form to their active form in a single step, or they may have one or more intermediate forms that may be active or inactive on their own. The preparation and uses of prodrugs are discussed in the following references: T. Higuchi and V. Stella, “Pro-drugs as Novel Delivery Systems,” Volume 14 of the A.C.S. Symposium Series, “Bioreversible Carriers in Drug Design,” edited by Edward B. Roche, American Pharmaceutical Association (API). The references cited hereby are incorporated in full. (References cited here are: *The Harmaceutical 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.)

As used herein, the term "soft drug" refers to a compound that exerts a pharmacological effect but breaks down into inactive metabolites and degradation products, resulting in a limited duration of activity. See, for example, "Soft drugs: Principles and methods for the design of safe drugs," Nicholas Bodor, *Medicinal Research Reviews*, Vol. 4, No. 4, pp. 449-469, 1984, which is hereby incorporated in full.

As used herein, the term "metabolite," such as an active metabolite, overlaps with the prodrug described above. Therefore, such a metabolite is a pharmacologically active compound or a compound further metabolized into a pharmacologically active compound, which is a derivative produced by metabolic processes within the subject's body. For example, such a metabolite can be produced by oxidation, reduction, hydrolysis, amidation, deamidation, esterification, deesterification, enzymatic cleavage, etc., of the administered compound or its salt or prodrug. An active metabolite is such a pharmacologically active derivative compound. For a prodrug, the prodrug compound is typically inactive or less active than the metabolite. For an active metabolite, the parent compound can be an active compound or an inactive prodrug.

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

As used herein, the term "pharmaceutically acceptable" means that a substance or composition is chemically and/or toxicologically compatible with the other components constituting the formulation and/or the subject being treated.

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

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

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

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

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

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

As used herein, the term "solvent" or "solventized form" refers to a solvation form containing stoichiometric or non-stoichiometric amounts of a solvent. Some compounds have a tendency to trap a fixed molar ratio of solvent molecules in a crystalline solid state, thereby forming a solvate. If the solvent is water, the solvate formed is a hydrate, and if the solvent is an alcohol, the solvate formed is an alcoholate. Hydrates are formed by one or more water molecules retaining their molecular state as _H₂O and combining with a molecule of a substance. Examples of solvents that form solvates include, but are not limited to, water, isopropanol, ethanol, methanol, DMSO, ethyl acetate, acetic acid, and ethanolamine.

As used herein, the terms "crystalline form," "polycrystalline form," "polymorph," and "polymorph" are used interchangeably and refer to the crystalline structures in which a compound (or its salts or solvates) can crystallize in different crystalline arrangements, all of which have the same elemental composition. Different crystalline forms typically exhibit different X-ray diffraction patterns, infrared spectra, melting points, densities, hardness, crystal shapes, optical and electrical properties, stability, and solubility. Recrystallization solvents, crystallization rates, storage temperatures, and other factors may cause one crystalline form to dominate. Polymorphs of a compound can be prepared by crystallization under different conditions.

Depending on the choice of substituents, the compounds of this disclosure may include one or more asymmetric centers and therefore may be in various stereoisomeric forms, such as enantiomers and/or diastereomers. For example, the compounds provided herein may have an asymmetric carbon center, and therefore the compounds provided herein may have (R) or (S) stereoconfigurations at the carbon asymmetric center. Thus, the compounds of this disclosure may be in the form of individual enantiomers, diastereomers, or geometric isomers, or may be in the form of mixtures of stereoisomers.

As used herein, the term "enantiomer" refers to two stereoisomers of a compound that are non-overlapping mirror images of each other. The term "diastereomer" refers to a pair of optical isomers that are not mirror images of each other. Diastereomers have different physical properties, such as melting point, boiling point, spectral properties, and reactivity.

When a particular enantiomer is preferred, in some embodiments it can be provided substantially free of the relative enantiomer and may also be referred to as “optically enriched.” As used herein, “optically enriched” means that the compound consists of a significantly larger proportion of one enantiomer. In some embodiments, the compound consists of at least about 90 wt% of the preferred enantiomer. In other embodiments, the compound consists of at least about 95 wt%, 98 wt%, or 99 wt% of the preferred enantiomer. The preferred enantiomer can be isolated from the racemic mixture by any method known to those skilled in the art, such as by chromatography or crystallization, by synthesis using stereochemically homogeneous starting materials, or by stereoselective synthesis. Optionally, derivatization can be performed prior to the separation of the stereoisomers. The separation of the mixture of stereoisomers can be performed as an intermediate step during the synthesis of the compound provided herein or can be performed on the final racemic product. The absolute stereochemistry can be determined by X-ray crystallography of the crystalline product or crystalline intermediate, which, if necessary, is derivatized with a reagent containing a stereocenter of known configuration. Alternatively, absolute stereochemistry can be determined by vibrational circular dichroism (VCD) spectroscopy. See, for example, Jacques et al., Enantiomers, Racemates and Resolutions (Wiley Interscience, New York, 1981); Wilen, S.H. et al., Tetrahedron 33:2725 (1977); Eliel, E.L., Stereochemistry of Carbon Compounds (McGraw-Hill, NY, 1962); Wilen, S.H., Tables of Resolving Agents and Optical Resolutions, p. 268 (E.L. Eliel, ed., Univ. of Notre Dame Press, IN, 1972).

In some embodiments, a mixture of diastereomers is provided, such as a mixture rich in 51% or more of one of the diastereomers, including, for example, 60% or more, 70% or more, 80% or more, or 90% or more of one of the diastereomers.

In some embodiments, unless otherwise indicated, the compounds provided herein may have one or more double bonds present as Z or E isomers. Additionally, this disclosure covers compounds in the form of a single isomer substantially free of other isomers, and alternatively, in the form of a mixture of multiple isomers (e.g., a racemic mixture of enantiomers).

The compounds disclosed herein may also exist in different tautomeric forms, and all such forms are covered within the scope of this disclosure. The terms "tautomer" or "tautomeric form" refer to structural isomers of different energies that can interconvert through low energy barriers. For example, proton tautomers (also called proton-heterotautomers) include interconversions via proton migration, such as keto-enol, amide-imino, lactam-lactamimide, imine-enamine isomerization, and cyclic forms where protons can occupy two or more positions in a heterocyclic system (e.g., 1H- and 3H-imidazolium, 1H-, 2H- and 4H-1,2,4-triazole, 1H- and 2H-isoindole, and 1H- and 2H-pyrazole). Valence tautomers include interconversions via the recombination of some bonding electrons. Tautomers may be in equilibrium or spatially locked into one form through appropriate substitution. Unless otherwise stated, compounds identified by name or structure as a particular tautomer form in this disclosure are intended to include other tautomer forms.

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

Compound Synthesis

The synthetic schemes described in the examples illustrate the synthesis of the compounds provided herein, including their pharmaceutically acceptable salts. The compounds provided herein can be prepared using any known organic synthetic technique and can be synthesized according to any of a variety of possible synthetic routes; therefore, these schemes are merely illustrative and not intended to limit other possible methods that can be used to prepare the compounds provided herein. Furthermore, the steps in these schemes are for better illustration and may be modified as needed. The embodiments of the compounds in the examples were synthesized for research purposes and possibly for submission to regulatory agencies.

The reactions for preparing the compounds of this disclosure can be carried out in suitable solvents that can be readily selected by those skilled in the art of organic synthesis. Suitable solvents are substantially non-reactive to the starting materials (reactants), intermediates, or products at temperatures in which the reaction takes place (e.g., temperatures ranging from the freezing point to the boiling point of the solvent). A given reaction can be carried out in one solvent or a mixture of more than one solvent. Depending on the specific reaction step, a suitable solvent for that particular reaction step can be selected by those skilled in the art.

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

The reaction can be monitored using any suitable method known in the art. For example, product formation can be monitored by spectroscopic means such as nuclear magnetic resonance spectroscopy (e.g., ^¹H or ^¹³C ), infrared spectroscopy, spectrophotometry (e.g., UV-Vis), mass spectrometry, or by chromatographic methods such as high-performance liquid chromatography (HPLC), liquid chromatography-mass spectrometry (LCMS), or thin-layer chromatography (TLC). Those skilled in the art can purify the compound using 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), 874-883, which is incorporated herein by reference in its entirety) and normal-phase silica gel chromatography.

The known starting materials disclosed herein can be synthesized using or according to methods known in the art, or can be purchased from commercial suppliers. Unless otherwise stated, analytical grade solvents and commercially available reagents are used without further purification.

Unless otherwise stated, all reactions disclosed herein are carried out under positive pressure of nitrogen or argon or in anhydrous solvent using a drying tube, and reaction flasks are typically fitted with rubber septa for introduction of substrates and reagents via syringe. Glassware is oven-dried and/or heated to dry.

For illustrative purposes, the following examples partially illustrate synthetic pathways for preparing the compounds of this disclosure and key intermediates. Those skilled in the art will understand that other synthetic pathways can be used to synthesize the compounds of this invention. Although specific starting materials and reagents are described, they can be readily substituted with other starting materials and reagents to provide a variety of derivatives and/or reaction conditions. Furthermore, many of the compounds prepared by the methods described below can be further modified according to this disclosure using conventional chemical methods well known to those skilled in the art.

Pharmaceutical Composition

In another aspect, pharmaceutical compositions are provided comprising one or more molecules or compounds disclosed herein or pharmaceutically acceptable salts thereof.

On the other hand, a pharmaceutical composition is provided comprising one or more molecules or compounds disclosed herein or pharmaceutically acceptable salts thereof, and at least one pharmaceutically acceptable excipient.

As used herein, the term "pharmaceutical composition" refers to a formulation of the present disclosure containing a molecule or compound in a form suitable for administration to a subject.

As used herein, the term "pharmaceuticalally acceptable excipient" means an excipient that can be used to prepare pharmaceutical compositions that are generally safe, non-toxic, and biologically and otherwise desirable, and includes excipients that are acceptable for both veterinary and human pharmaceutical use. As used herein, "pharmaceuticalally acceptable excipient" includes one or more such excipients. The term "pharmaceuticalally acceptable excipient" also covers "pharmaceuticalally acceptable carriers" and "pharmaceuticalally acceptable diluents."

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

In some embodiments, suitable excipients may include buffers such as phosphates, citrates, and other organic acids; antioxidants, including ascorbic acid and methionine; preservatives (such as octadecyl dimethyl benzyl ammonium chloride; hexamethyl ammonium chloride; benzalkonium chloride; benzyl chloride; phenol, butanol, or benzyl alcohol; alkyl esters of p-hydroxybenzoate, such as methyl or propyl p-hydroxybenzoate; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) peptides; and proteins. Such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers, such as polyvinylpyrrolidone; amino acids, such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates, including glucose, mannose, or dextran; 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 nonionic surfactants, such as TWEEN ^™ , PLURONICS ^™ , or polyethylene glycol (PEG).

In some embodiments, suitable excipients may include one or more stabilizers, surfactants, wetting agents, lubricants, emulsifiers, suspending agents, preservatives, antioxidants, opacifiers, flow aids, processing aids, colorants, sweeteners, flavorings, and other known additives to provide an optimal presentation of the drug (i.e., the compounds of this disclosure or pharmaceutical compositions thereof) or to aid in the manufacture of a pharmaceutical product (i.e., a drug). The active pharmaceutical ingredient may also be encapsulated in microcapsules, for example, prepared by coagulation techniques or by interfacial polymerization, such microcapsules being, for example, hydroxymethyl cellulose or gelatin microcapsules and poly-(methyl methacrylate) microcapsules in colloidal drug delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles, and nanocapsules) or in crude emulsions. Such techniques are disclosed in Remington’s Pharmaceutical Sciences, 16th edition, Osol, A. (1980). “Liposomes” are small vesicles comprising various types of lipids, phospholipids, and/or surfactants that can be used to deliver drugs (such as the compounds disclosed herein and optionally chemotherapeutic agents) to mammals, including humans. The components of liposomes are usually arranged in a bilayer, similar to the lipid arrangement of biological membranes.

The pharmaceutical compositions provided herein may be administered to subjects, including but not limited to humans, and may be formulated into any form compatible with the intended route of administration.

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

The amount of active ingredient in a unit dosage form of the composition is the therapeutically effective amount and varies depending on the specific treatment involved. As used herein, the term "therapeuticly effective amount" refers to the amount of a molecule, compound, or composition containing said molecule or compound that treats, improves, or prevents an identified disease or symptom or exhibits detectable therapeutic or inhibitory effects. The effect can be detected by any assay method known in the art. The precise effective amount for a subject will depend on the subject's weight, body size, and health status; the nature and severity of the symptom; the rate of administration; the chosen treatment or combination of treatments for administration; and the prescribing physician's judgment. The therapeutically effective amount in a given situation can be determined through routine testing within the skill and judgment of a clinician.

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

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

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

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

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

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

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

In some embodiments, the pharmaceutical compositions disclosed herein may be in the form of an injectable formulation.

In some embodiments, the pharmaceutical compositions of this disclosure may be in the form of sterile injectable formulations, such as sterile injectable aqueous or oily suspensions. Such suspensions may be formulated using suitable dispersants or wetting agents and suspending agents mentioned above, according to known techniques. Sterile injectable formulations may also be sterile injectable solutions or suspensions in non-toxic parenteral diluents or solvents, such as solutions in 1,3-butanediol, or prepared as lyophilized powders. Acceptable mediators and solvents that may be used include water, Ringer's solution, and isotonic sodium chloride solution. Additionally, sterile non-volatile oils are conventionally used as solvents or suspension media. For this purpose, any mild non-volatile oil may be used, including synthetic monoglycerides or diglycerides. Furthermore, fatty acids such as oleic acid may also be used in the preparation of injectable formulations.

In some embodiments, the pharmaceutical compositions disclosed herein may be in the form of an inhalation formulation.

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

In some embodiments, the pharmaceutical compositions of this disclosure may be in the form of a topical or transdermal formulation.

In some embodiments, the pharmaceutical compositions provided herein may be in the form of creams, ointments, gels, and aqueous or oily solutions or suspensions, which are typically obtained by formulating the active ingredient with conventional, locally acceptable excipients, such as animal and vegetable fats, oils, waxes, paraffins, starch, astragalus gum, cellulose derivatives, polyethylene glycol, silicone, bentonite, silicic acid, talc, and zinc oxide, or mixtures thereof.

In some embodiments, the pharmaceutical compositions provided herein may be formulated in the form of transdermal patches, as is well known to those skilled in the art.

In addition to the representative dosage forms described above, pharmaceutically acceptable excipients and carriers are generally known to those skilled in the art and are therefore included in this disclosure. Such excipients and carriers are described, for example, in the following references: Remington Pharmaceutical Sciences, Mark Publishing, NJ (1991); Remington: The Science and Practice of Pharmacy, edited by University of the Sciences in Philadelphia, 21st edition, LWW (2005), which are incorporated herein by reference.

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

In some embodiments, the pharmaceutical compositions of this disclosure can be formulated to be administered at doses ranging from 0.001 mg/kg body weight/day to 1000 mg/kg body weight/day, for example, 0.01 mg/kg body weight/day to 800 mg/kg body weight/day, 0.01 mg/kg body weight/day to 700 mg/kg body weight/day, 0.01 mg/kg body weight/day to 600 mg/kg body weight/day, 0.01 mg/kg body weight/day to 500 mg/kg body weight/day, 0.01 mg/kg body weight/day to 400 mg/kg body weight/day, 0.01 mg/kg body weight/day to 300 mg/kg body weight/day, 0.1 mg/kg body weight/day to 200 mg/kg body weight/day, 0.1 mg/kg body weight/day to 150 mg/kg body weight/day, 0.1 mg/kg body weight/day, etc. The compounds provided herein or their pharmaceutically acceptable salts are available in doses ranging from 0.5 mg/kg body weight/day to 100 mg/kg body weight/day, 0.5 mg/kg body weight/day to 80 mg/kg body weight/day, 0.5 mg/kg body weight/day to 60 mg/kg body weight/day, 0.5 mg/kg body weight/day to 50 mg/kg body weight/day, 1 mg/kg body weight/day to 50 mg/kg body weight/day, 1 mg/kg body weight/day to 45 mg/kg body weight/day, 1 mg/kg body weight/day to 40 mg/kg body weight/day, 1 mg/kg body weight/day to 35 mg/kg body weight/day, 1 mg/kg body weight/day to 30 mg/kg body weight/day, and 1 mg/kg body weight/day to 25 mg/kg body weight/day. In some cases, dose levels below the lower limit of the aforementioned range may be sufficient, while in others, larger doses may be used without causing any harmful side effects, provided that the larger dose is first divided into several smaller doses for administration throughout the day. For more information on routes of administration and dosage regimens, see Comprehensive Medicinal Chemistry, Volume 5, Chapter 25.3 (Corwin Hansch; Chairman of the Editorial Board), Pergman Publishers, 1990, the references of which are specifically incorporated herein by reference.

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

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

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

In another aspect, a pharmaceutical composition is provided comprising one or more compounds of the present disclosure as a first active ingredient or a pharmaceutically acceptable salt thereof, and a second active ingredient.

In some embodiments, the second active ingredient has an activity complementary to that of the compounds provided herein, such that they do not adversely affect each other. Such ingredients are suitably present in a combination of amounts effective for the intended purpose.

In some embodiments, the second active ingredient may include:

(i) Antiproliferative/anti-proliferative agents and combinations thereof used in medical oncology, such as alkylating agents (e.g., cisplatin, carboplatin, cyclophosphamide, nitrogen mustard, melphalan, chlorambucil, busulphan, and nitrosourea); antimetabolites (e.g., antifolate agents such as fluoropyrimidines (like 5-fluorouracil and tegafur), raltitrexed, methotrexate, cytarabine, hydroxyurea, and gemcitabine); and antitumor antibiotics (e.g., anthracyclines such as adriamycin, bleomycin, doxorubicin, daunomycin, epirubicin). irubicin, idarubicin, mitomycin-C, dactinomycin, and mithramycin; antimitotic agents (e.g., vinca alkaloids, such as vincristine, vinblastine, vindesine, and vinorelbine) and taxanes (e.g., paclitaxel and taxotere); and topoisomerase inhibitors (e.g., epipodophyllotoxin, such as etoposide and teniposide), amsacrine, topotecan, and camptothecins);

(ii) Cell growth inhibitors, such as anti-estrogens (e.g., tamoxifen, toremifene, raloxifene, droloxifene, and iodoxyfene), estrogen receptor downregulators (e.g., fulvestrant), and anti-androgens (e.g., bicalutamide, flutamide, nilutamide, and acetylcholine). LHRH antagonists or agonists (e.g., goserelin, leuprorelin, and buserelin), progestins (e.g., megestrol acetate), aromatase inhibitors (e.g., anastrozole, letrozole, vorazole, and exemestane), and 5α-reductase inhibitors (e.g., finasteride).

(iii) Anti-invasive agents (e.g., c-Src kinase family inhibitors, such as 4-(6-chloro-2,3-methylenedioxyanilino)-7-[2-(4-methylpiperazin-1-yl)ethoxy]-5-tetrahydropyran-4-yloxyquinazoline (AZD0530) and N-(2-chloro-6-methylphenyl)-2-{6-[4-(2-hydroxyethyl)piperazin-1-yl]-2-methylpyrimidin-4-ylamino}thiazolyl-5-carboxamide (dasatinib, BMS-354825)) and metalloproteinase inhibitors, such as marimastat and urokinase plasminogen activator receptor function inhibitors);

(iv) Inhibitors of growth factor function: For example, such inhibitors include growth factor antibodies and growth factor receptor antibodies (e.g., anti-erbB2 antibody trastuzumab [Herceptin ^™ ] and anti-erbB1 antibody cetuximab [C225]); such inhibitors also include, for example, tyrosine kinase inhibitors, such as epidermal growth factor family inhibitors (e.g., EGFR family tyrosine kinase inhibitors, such as N-(3-fluoro-4-fluorophenyl)-7-methoxy-6-(3-morpholinopropoxy)quinazoline-4-amine (gefitinib, ZD...). 1839), N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)quinazoline-4-amine (erlotinib, OSI-774) and 6-acrylamido-N-(3-fluoro-4-fluorophenyl)-7-(3-morpholinopropoxy)quinazoline-4-amine (CI 1033) and erbB2 tyrosine kinase inhibitors, such as lapatinib, hepatocyte growth factor family inhibitors, platelet-derived growth factor family inhibitors, such as imatinib, serine/threonine kinase inhibitors (e.g., Ras/Raf signaling inhibitors, such as farnesyltransferase inhibitors, such as sorafenib (BAY 43-9006)), and cell signaling inhibitors via MEK and/or Akt kinases;

(v) Anti-angiogenic agents, such as anti-angiogenic agents that inhibit the action of vascular endothelial growth factor, [e.g., the anti-vascular endothelial growth factor antibody bevacizumab (Avastin ^™ ) and VEGF receptor tyrosine kinase inhibitors, such as 4-(4-bromo-2-fluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazoline (ZD6474; Example 2 in WO01/32651), 4-(4-fluoro-2-methylindole-5-yloxy)-6-methoxy-7-(3-pyrrolidine-1-ylpropoxy)quinazoline (AZD2171; Example 240 in WO00/47212), vatalanib (PTK787; WO98/35985), and SU11248 (sunitinib; WO00/35985). 01/60814), and compounds that act through other mechanisms (e.g., linomide, integrin ανβ3 function inhibitors, and angiostatin);

(vi) Vascular damaging agents, such as comprehending A4 and compounds disclosed in international patent applications WO 99/02166, WO 00/40529, WO 00/41669, WO 01/92224, WO 02/04434 and WO 02/08213;

(vii) Antisense therapy, such as ISIS 2503, an anti-ras antisense agent;

(viii) Gene therapy approaches, including, for example, methods of replacing abnormal genes, such as abnormal p53 or abnormal BRC1 or BRCA2; GDEPT (gene-directed enzyme prodrug therapy) methods, such as those using cytosine deaminase, thymidine kinase, or bacterial nitroreductase; and methods of increasing patient tolerance to chemotherapy or radiotherapy, such as multidrug-resistant gene therapy; and

(ix) Immunotherapy methods include in vitro and in vivo methods to increase the immunogenicity of a patient’s tumor cells (e.g., transfection with cytokines such as interleukin-2, interleukin-4, or granulocyte-macrophage colony-stimulating factor), methods to reduce T-cell dysfunction, methods using transfected immune cells (e.g., dendritic cells transfected with cytokines), methods using tumor cell lines transfected with cytokines, and methods using anti-idiotype antibodies.

Methods of treating diseases

On the one hand, this disclosure provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, which is capable of inhibiting ATR kinase. The inhibitory properties of the compound of formula (I) can be demonstrated using the test procedures described herein.

Therefore, compounds of formula (I) can be used to treat (therapeutic or preventative) symptoms or diseases mediated by ATR kinases in subjects.

As used herein, "subject" refers to both humans and non-human animals. Examples of non-human animals include all vertebrates, such as mammals, including non-human primates (especially higher primates), dogs, rodents (e.g., mice or rats), guinea pigs, and cats; and non-mammals, such as birds, amphibians, reptiles, etc. In a preferred embodiment, the subject is a human. In another embodiment, the subject is a laboratory animal or an animal suitable as a disease model.

In some embodiments, the compound of formula (I) can be used as an antitumor agent. In some embodiments, the compound of formula (I) can be used as an antiproliferative, apoptotic, and/or antiinvasive agent to inhibit and/or treat solid and/or liquid tumor diseases. In some embodiments, the compound of formula (I) can be used to prevent or treat those tumors sensitive to ATR inhibition. In some embodiments, the compound of formula (I) can be used to prevent or treat those tumors mediated solely or partially by ATR.

In some embodiments, the compound of formula (I) can be used to treat proliferative diseases, including malignant diseases (such as cancer) and non-malignant diseases such as inflammatory diseases, obstructive airway diseases, immune diseases or cardiovascular diseases.

In some embodiments, the compound of formula (I) can be used to treat cancers, such as, but not limited to, hematologic malignancies (e.g., leukemia, multiple myeloma), lymphomas (e.g., Hodgkin's disease, non-Hodgkin's lymphomas (including mantle cell lymphoma)), and myelodysplastic syndromes, as well as solid tumors and their metastases (e.g., breast cancer, lung cancer (non-small cell lung cancer (NSCL), small cell lung cancer (SCLC), squamous cell carcinoma), endometrial cancer), central nervous system tumors (e.g., glioma, dysembryonic neuroepithelial tumor, glioblastoma multiforme, mixed glioma, medulloblastoma, retinoblastoma, etc.). Cancers include: blastoma, germ cell tumor, and teratoma; gastrointestinal cancers (such as stomach cancer, esophageal cancer, hepatocellular carcinoma, bile duct cancer, colon and rectal cancer, small bowel cancer, and pancreatic cancer); skin cancers (such as melanoma (especially metastatic melanoma)); thyroid cancer; head and neck cancer and salivary gland cancer; prostate cancer; testicular cancer; ovarian cancer; cervical cancer; uterine cancer; vaginal cancer; bladder cancer; kidney cancer (including renal cell carcinoma, clear cell carcinoma, and renal eosinophilic tumor)); squamous cell carcinoma; sarcomas (such as osteosarcoma, chondrosarcoma, leiomyosarcoma, soft tissue sarcoma, Ewing's sarcoma, gastrointestinal stromal tumor (GIST), Kaposi's sarcoma); and pediatric cancers (such as rhabdomyosarcoma and neuroblastoma).

In some embodiments, the compound of formula (I) can be used to treat autoimmune and/or inflammatory diseases, such as, but not limited to, allergies, Alzheimer's disease, acute disseminated encephalomyelitis, Addison's disease, ankylosing spondylitis, antiphospholipid antibody syndrome, asthma, atherosclerosis, autoimmune hemolytic anemia, autoimmune hemolytic and thrombocytopenic states, autoimmune hepatitis, autoimmune inner ear diseases, bullous pemphigoid, celiac disease, chagas disease, chronic obstructive pulmonary disease, chronic idiopathic thrombocytopenic purpura (ITP), Chaug-Strauss syndrome, Crohn's disease, dermatomyositis, type 1 diabetes, endometriosis, Goodpasture's syndrome (and associated glomerulonephritis and pulmonary hemorrhage). Graves' disease, Guillain-Barré syndrome, Hashimoto's disease, hidradenitis suppurativa, idiopathic thrombocytopenic purpura, interstitial cystitis, irritable bowel syndrome, lupus erythematosus, scleroderma, multiple sclerosis, myasthenia gravis, somnolence, neuromuscular rigidity, Parkinson's disease, pemphigus vulgaris. Pernicious anemia, polymyositis, primary biliary cirrhosis, psoriasis, psoriatic arthritis, rheumatoid arthritis, schizophrenia, septic shock, scleroderma, Sjogren's disease, systemic lupus erythematosus (and associated glomerulonephritis), temporal arteritis, tissue transplant rejection and hyperacute rejection of transplanted organs, vasculitis (ANCA-associated vasculitis and other vasculitis), vitiligo, and Wegener's granulomatosis.

As used herein, the term "therapy" is intended to have its normal meaning, namely, treating a disease to completely or partially alleviate one, some, or all of its symptoms, or to correct or compensate for the underlying pathology, thereby achieving a beneficial or desired clinical outcome. For the purposes of this disclosure, beneficial or desired clinical outcomes include, but are not limited to, symptom relief, reduction of disease severity, stabilization of the disease state (i.e., non-deterioration), delay or slowing of disease progression, improvement or mitigation of the disease state, and remission (whether partial or complete), whether detectable or undetectable. "Therapy" may also mean prolonged survival compared to expected survival without therapy. Situations requiring therapy include having a symptom or condition, being susceptible to a symptom or condition, or needing to prevent a symptom or condition. Unless specifically indicated to the contrary, the term "therapy" also encompasses prevention. The terms "therapeutic" and "therapeutically" should be interpreted accordingly.

As used herein, the term “prophylaxis” or “prophylactic” is intended to have its normal meaning and includes primary prevention for preventing the development of disease and secondary prevention for protecting patients who have already developed disease from the onset or worsening of the disease or the development of new disease-related symptoms, either temporarily or permanently.

The term “treatment” is used synonymously with “therapeutic therapy.” Similarly, the term “treatment” can be considered as “the application of a therapeutic therapy,” where “therapeutic therapy” is as defined herein.

In another aspect, this disclosure provides the use of a compound of the disclosure or a pharmaceutically acceptable salt thereof or a pharmaceutical composition thereof for therapeutic purposes, such as for a therapy related to the ATR protein.

In another aspect, this disclosure provides the use of a compound of the disclosure or a pharmaceutically acceptable salt thereof or a pharmaceutical composition thereof in the preparation of a medicament for treating cancer.

In another aspect, this disclosure provides the use of a compound of the disclosure or a pharmaceutically acceptable salt thereof or a pharmaceutical composition thereof in the preparation of a medicament for treating cancer.

On the other hand, this disclosure provides compounds of the present disclosure or pharmaceutically acceptable salts thereof or pharmaceutical compositions thereof for the treatment of cancer.

In some embodiments, the compound of formula (I) may be further used in combination with other bioactive ingredients (such as, but not limited to, second and different anti-vesicle agents) and non-pharmacological therapies (such as, but not limited to, surgery or radiation therapy). For example, the compound of formula (I) may be used in combination with other pharmaceutically active compounds or non-pharmacological therapies, preferably compounds capable of enhancing the effect of the compound of formula (I). The compound of formula (I) may be administered simultaneously (as a single formulation or a separate formulation) or sequentially with other therapies. Typically, combination therapy envisions the administration of two or more drugs/therapies during a single cycle or course of treatment.

In some embodiments, the compound of formula (I) is used in combination with one or more conventional chemotherapeutic agents, which cover a wide range of therapeutic treatments in the field of oncology. These agents are administered at various stages of the disease to shrink tumors, destroy residual cancer cells left after surgery, induce remission, maintain remission, and/or alleviate symptoms associated with cancer or its treatment.

In some embodiments, the compound of formula (I) is used in combination with one or more targeted anticancer agents that regulate protein kinases involved in various disease states.

In some embodiments, the compound of formula (I) is used in combination with one or more targeted anticancer agents that regulate non-kinase biological targets, pathways or processes.

In some embodiments, the compound of formula (I) is used in combination with one or more other anticancer agents, including but not limited to gene therapy, RNAi cancer therapy, chemopreservatives (e.g., amfostine, mesna, dexrazoxane), antibody conjugates (e.g., brentuximab vedotin, ibritumomab tioxetan), cancer immunotherapy (e.g., interleukin-2), cancer vaccines (e.g., sipuleucel-T), or monoclonal antibodies (e.g., bevacizumab, alemtuzumab, rituximab, trastuzumab, etc.).

In some embodiments, the compound of formula (I) is used in combination with one or more anti-inflammatory agents, including but not limited to NSAIDs, non-specific and COX-2 specific cyclooxygenase inhibitors, gold compounds, corticosteroids, methotrexate, tumor necrosis factor receptor (TNF) receptor antagonists, immunosuppressants, and methotrexate.

In some embodiments, the compound of formula (I) is used in combination with radiotherapy or surgery. Radiation is typically delivered from inside the body (by implanting radioactive material near the cancer site) or from outside the body by a machine that uses photons (X-rays or gamma rays) or particle radiation. When the combination therapy further includes radiotherapy, radiotherapy can be administered at any suitable time, as long as the beneficial effects of the combination of the therapeutic agent and radiotherapy are achieved.

Therefore, in another aspect, this disclosure provides a method for treating an ATR kinase-related disease in a subject of need, the method comprising administering to the subject an effective amount of a compound of this disclosure or a pharmaceutically acceptable salt thereof or a pharmaceutical composition of this disclosure.

Example

The following examples are included for illustrative purposes. However, it should be understood that these examples do not limit the present disclosure and are intended only to illustrate the methods of practicing the present disclosure. Those skilled in the art will recognize that the described chemical reactions can be readily adapted to prepare a variety of other compounds of the present disclosure, and alternative methods for preparing the compounds of the present disclosure are considered to be within the scope of the present disclosure. For example, non-exemplary compounds according to the present disclosure can be successfully synthesized by modifications obvious to those skilled in the art, such as by appropriately protecting interfering groups, by utilizing other suitable reagents and building blocks known in the art besides those described, and/or by conventional modifications to the reaction conditions. Alternatively, other reactions disclosed herein or known in the art will be considered suitable for preparing other compounds of the present disclosure.

Example 1

Step 1: (R)-2-chloro-6-(3-methylmorpholino)isonicotinic acid methyl ester (1-3)

_Cs₂CO₃ ( _7.91 g, 24.27 mmol) and Pd(dppf)Cl₂ (0.44 g, 0.61 mmol) were added to a solution of methyl 2,6-dichloropyridine-4-carboxylate (2.5 g, 12.13 mmol) and (3R) _-3 -methylmorpholine (1.35 g, 13.35 mmol) in dioxane (50 mL). The mixture was injected twice with _N₂ and stirred overnight at 100 °C. LC-MS showed the reaction was complete. After cooling to room temperature, the reaction mixture was diluted with EA (80 mL), washed with water and brine, _dried over anhydrous _Na₂SO₄ , filtered, and concentrated. The residue was purified by rapid column chromatography (silica gel, 0–15% ethyl acetate/petroleum ether) to give the desired product (1.01 g, yield: 31%). LC-MS(ESI):m/z 271[M+H] ⁺ . ¹ H NMR (400MHz, DMSO) δ7.11(d,J＝0.7Hz,1H),7.00(d,J＝0.7Hz,1H),4.32(dd,J＝6.7,2.6Hz,1H),3.96–3.88(m,2H),3.87(s,3H),3.72 (d, J＝11.4Hz, 1H), 3.61 (dd, J＝11.5, 3.0Hz, 1H), 3.46 (td, J＝11.9, 3.1Hz, 1H), 3.13 (td, J＝12.7, 3.9Hz, 1H), 1.15 (d, J＝6.7Hz, 3H).

Step 2. (R)-(2-chloro-6-(3-methylmorpholino)pyridin-4-yl)methanol (1-4)

At 0 °C, a solution of _LiBH₄ (2.0 M in THF, 15.0 mL, 30.0 mmol) was added to a solution of methyl 2-chloro-6-[(3R)-3-methylmorpholin-4-yl]pyridine-4-carboxylate (4.5 g, 16.62 mmol) in THF (40 mL). The mixture was stirred overnight at room temperature under a nitrogen atmosphere. LC-MS showed that the reaction was complete. The reaction mixture was quenched with a saturated aqueous solution _{of NaHCO₃} and extracted with EA (60 mL × 2). The combined organic layers were washed with brine, dried over _{anhydrous Na₂SO₄} , filtered, and concentrated under vacuum. The residue was purified by rapid column chromatography (silica gel, 0–50% ethyl acetate/petroleum ether) to give the title product (3.87 g, 96%). LC-MS(ESI): m/z 243[M+H] ⁺ . ¹ H NMR (400MHz, CDCl ₃ )δ6.58(s,1H),6.46(s,1H),4.62(s,2H),4.31–4.23(m,1H),3.99(dd,J＝11.4,3.8Hz,1H),3.86(dd,J＝13.1,2.9Hz,1H),3.78(d, J＝11.3Hz,1H),3.72(dd,J＝11.4,2.9Hz,1H),3.61–3.54(m,1H),3.21(td,J＝12.7,3.8Hz,1H),1.89(s,1H),1.24(d,J＝6.7Hz,3H).

Step 3. (R)-4-(6-chloro-4-(chloromethyl)pyridin-2-yl)-3-methylmorpholine (1-5)

SOCl₂ (10 mL, 137.8 mmol) was added dropwise to a solution of {2-chloro-6-[(3R)-3-methylmorpholin-4-yl]pyridin-4-yl}methanol (4.0 g, 16.48 mmol) and DMF (0.05 mL, 0.65 mmol) in DCM (40 mL) at ₀ °C. The resulting mixture was stirred at room temperature under a nitrogen atmosphere for 1 hour. LC-MS showed that the reaction was complete. The reaction mixture was concentrated to dryness under vacuum. The residue was dissolved in DCM (50 mL), washed with a saturated aqueous solution of _NaHCO₃ and brine, dried over anhydrous _{Na₂SO₄ ,} filtered, and concentrated to give the desired product (4.08 g, yield: 95%). LC-MS(ESI): m/z 261[M+H] ⁺ . ¹ H NMR (400MHz, CDCl ₃ )δ6.62(s,1H),6.43(s,1H),4.41(s,2H),4.26(dd,J＝6.7,2.6Hz,1H),4.00(dd,J＝11.4,3.8Hz,1H),3.86(dd,J＝13.1,3.0Hz, 1H), 3.80–3.76 (m, 1H), 3.73 (dd, J＝11.4, 2.9Hz, 1H), 3.61–3.54 (m, 1H), 3.22 (td, J＝12.7, 3.9Hz, 1H), 1.26 (d, J＝6.7Hz, 3H).

Step 4. (R)-4-(6-chloro-4-((methanesulfonyl)methyl)pyridin-2-yl)-3-methylmorpholine (1-6)

A mixture of (3R)-4-[6-chloro-4-(chloromethyl)pyridin-2-yl]-3-methylmorpholine (1.50 g, 5.74 mmol) and sodium methanesulfonate (1.17 g, 11.49 mmol) in DMF (20 mL) was stirred overnight at room temperature under a nitrogen atmosphere. _LC -MS showed that the reaction was complete. The reaction mixture was diluted with _H₂O and extracted with EA (60 mL × 2). The combined organic layers were washed with brine, dried over anhydrous _Na₂SO₄ , filtered, and concentrated. The residue was purified by rapid column chromatography (silica gel, 0–50% ethyl acetate/petroleum ether) to give the desired product (1.55 g, yield: 89%). LC-MS(ESI): m/z 305[M+H] ⁺ . ¹ H NMR (400MHz, CDCl ₃ )δ6.60(s,1H),6.49(s,1H),4.28–4.22(m,1H),4.10(s,2H),4.00(dd,J＝11.5,3.8Hz,1H),3.90(dd,J＝13.2,2.8Hz,1H),3.80– 3.76(m,1H),3.72(dd,J＝11.4,3.0Hz,1H),3.61–3.54(m,1H),3.23(td,J＝12.7,3.9Hz,1H),2.85(s,3H),1.26(d,J＝6.7Hz,3H).

Step 5. (R)-4-(6-chloro-4-(1-(methanesulfonyl)cyclopropyl)pyridin-2-yl)-3-methylmorpholine (1-7)

A mixture of (3R)-4-[6-chloro-4-(methanesulfonylmethyl)pyridin-2-yl]-3-methylmorpholine (1.55 g, 5.09 mmol), 1,2-dibromoethane (0.88 mL, 10.17 mmol), NaOH solution (10.0 M, 5.09 mL, 50.85 mmol), and TBAB (330 mg, 1.02 mmol) in toluene (50 mL) was stirred overnight at 60 °C under a nitrogen atmosphere. LC-MS showed the reaction was complete. The reaction mixture was diluted with _H₂O and extracted with EA (60 mL × 2). The combined organic layers were washed with brine, dried over _{anhydrous Na₂SO₄} , filtered, and concentrated under vacuum. The residue was purified by rapid column chromatography (silica gel, 0–50% ethyl acetate/petroleum ether) to give the desired product (652 mg, yield: 39%). LC-MS(ESI): m/z 331[M+H] ⁺ . ¹ H NMR (400MHz, CDCl ₃ )δ6.70(s,1H),6.67(d,J＝0.9Hz,1H),4.26(d,J＝6.9Hz,1H),4.00(dd,J＝11.4,3.8Hz,1H),3.89(dd,J＝13.2,2.8Hz,1H),3.78(d,J＝11.4Hz,1H), 3.72(dd,J＝11.4,3.0Hz,1H),3.58(td,J＝11.9,3.1Hz,1H),3.22(td,J＝ 12.7, 3.9Hz, 1H), 2.83 (s, 3H), 1.88–1.76 (m, 2H), 1.26 (d, J = 6.6Hz, 5H).

Step 6. (R)-5-((6-(3-methylmorpholino)-4-(1-(methanesulfonyl)cyclopropyl)pyridin-2-yl)amino)-1H-pyrazole-1-carboxylic acid tert-butyl ester (1-9)

A solution of (3R)-4-[6-chloro-4-(1-methanesulfonylcyclopropyl)pyridin-2-yl]-3-methylmorpholine (100 mg, 0.30 mmol) and tert-butyl 5-amino-1H-pyrazole-1-carboxylate (83 mg, 0.45 mmol) in dioxane (10 mL) was added with BrettPhos-Pd-G3 catalyst (27 mg, 0.030 mmol ₎ and _Cs₂CO₃ (197 mg, 0.060 mmol). The mixture was injected twice with _N₂ and stirred at 100 °C for 4 h. LC-MS showed the reaction was complete. The reaction mixture was diluted with EA (50 mL), washed with water and brine, dried over _{anhydrous Na₂SO₄} , filtered, and concentrated under vacuum. The residue was purified by rapid column chromatography (silica gel, 0–60% ethyl acetate/petroleum ether) to give the title product (59 mg, yield: 41%). LC-MS(ESI): m/z 478[M+H] ⁺ .

Step 7. (R)-6-(3-methylmorpholino)-4-(1-(methanesulfonyl)cyclopropyl)-N-(1H-pyrazol-5-yl)pyridin-2-amine (1)

At room temperature, a mixture of 5-{[4-(1-methanesulfonylcyclopropyl)-6-[(3R)-3-methylmorpholin-4-yl]pyridin-2-yl]amino}-1H-pyrazole-1-carboxylic acid tert-butyl ester (59 mg, 0.12 mol) and HCl solution (4 M in dioxane, 2 mL) in DCM (2 mL) was stirred for 2 hours. LC-MS showed the reaction was complete. The reaction mixture was concentrated to dryness under vacuum. The residue was purified by preparative HPLC (C18, 20-95%, acetonitrile in _H₂O containing 0.1% HCOOH) to give the desired product (14.2 mg, yield: 30%). LC-MS (ESI): m/z 378 [M+H] ⁺ . ¹ H NMR (400MHz, DMSO) δ9.14(s,1H),7.57(d,J＝2.2Hz,1H),6.60(s,1H),6.28(d,J＝2.2Hz,1 H),6.23(s,1H),4.26(d,J＝6.6Hz,1H),3.94(dd,J＝11.3,3.3Hz,1H),3.81(d,J＝13.0Hz, 1H),3.73(d,J＝11.2Hz,1H),3.65–3.61(m,1H),3.51(s,1H),3.08(d,J＝3.6Hz,1H),2.95 (s, 3H), 1.57 (dd, J = 5.8, 4.0 Hz, 2H), 1.26 ( dd, J = 6.3, 4.7 Hz, 2H), 1.14 ( d, J = 6.6 Hz, 3H).

Example 2

Step 1. (2,6-Dichloro-3-methylpyridin-4-yl)methanol (2-2)

At 0 °C under a _nitrogen atmosphere, _LiBH₄ solution (2.0 M in THF, 0.68 mL, 1.37 mmol) was added dropwise to a solution of ethyl 2,6-dichloro-3-methylpyridine-4-carboxylate (290 mg, 1.24 mmol) in anhydrous THF (5 mL). The resulting mixture was stirred at 0 °C for 1 hour. LC-MS showed that the reaction was complete. The reaction mixture was quenched with a saturated aqueous _NH₄Cl solution and extracted with EA (50 mL). The organic layer was dried over _{anhydrous Na₂SO₄} , filtered, and concentrated under vacuum. The residue was purified by silica gel column chromatography (PE:EA = 5:1, V/V) to give the desired product (220 mg, yield: 92%). LC/MS (ESI): m/z 192 [M+H] ^⁺ .

Step 2.2, 6-Dichloro-4-(chloromethyl)-3-methylpyridine (2-3)

_SOCl₂ (408 mg, 3.44 mmol) was added dropwise to a solution of (2,6-dichloro-3-methylpyridin-4-yl)methanol (220 mg, 1.14 mmol) and DMF (0.01 mL) in anhydrous DCM (5 mL) at 0 °C. The resulting mixture was stirred at room temperature for 1 hour. LC-MS showed that the reaction was complete. The reaction mixture was concentrated under reduced pressure. The residue was diluted with EA (40 mL), washed with saturated _aqueous NaHCO₃ solution and brine, dried over _{anhydrous Na₂SO₄} , filtered, and concentrated under vacuum. The residue was used for the next step without further purification (230 mg, yield: 95%). LC/MS (ESI): m/z 210/212 [M+H] ^⁺ .

Step 3.2, 6-Dichloro-3-methyl-4-((methanesulfonyl)methyl)pyridinemorpholine (2-4)

A mixture of 2,6-dichloro-4-(chloromethyl)-3-methylpyridine (259 mg, 1.23 mmol) and _CH₃SO₂Na ( ₂₅₃ mg, 2.48 mmol) in DMF (5 mL) was stirred for 4 hours at room temperature. _LC -MS showed the reaction was complete. The reaction mixture was diluted with EA (40 mL), washed with water and brine, dried over anhydrous _Na₂SO₄ , filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 1:1, V/V) to give the desired product (270 mg, yield: 86%). LC/MS (ESI): m/z 254 [M+H] ^⁺ .

Step 4. (R)-4-(6-chloro-5-methyl-4-((methanesulfonyl)methyl)pyridin-2-yl)-3-methylmorpholine (2-6)

A mixture of 2,6-dichloro-4-(methanesulfonylmethyl)-3-methylpyridine (250 mg, 0.98 mmol), (3R)-3-methylmorpholine (399 mg, 3.94 mmol), and DIPEA (509 mg, 3.94 mmol) in NMP (3 mL) was stirred for 1 hour at 180 °C under microwave irradiation. _LC -MS showed the reaction was complete. The reaction mixture was diluted with EA (40 mL), washed with water and brine, dried over anhydrous _Na₂SO₄ , filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 1:1, V/V) to give the desired product (97 mg, yield: 30%) as a white solid. LC/MS (ESI): m/z 319 [M+H] ^⁺ .

Step 5. (R)-4-(6-chloro-5-methyl-4-(1-(methanesulfonyl)cyclopropyl)pyridin-2-yl)-3-methylmorpholine (2-7)

A mixture of (3R)-4-[6-chloro-4-(methanesulfonylmethyl)-5-methylpyridin-2-yl]-3-methylmorpholine (97 mg, 0.30 mmol), 1,2-dibromoethane (113 mg, 0.60 mmol), NaOH (10.0 M in _H₂O , 0.3 mL, 3.05 mmol), and TBAB (19 mg, 0.06 mmol) in toluene (4 mL) was stirred for 3 hours at 60 °C. LC-MS showed the reaction was complete. The reaction mixture was diluted with EA (40 mL), washed with water and brine, dried over _{anhydrous Na₂SO₄} , filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 1:1, V/V) to give the desired product (29 mg, yield: 27%). LC/MS (ESI): m/z 345 [M+H] ^⁺ .

Step 6. (R)-3-methyl-6-(3-methylmorpholino)-4-(1-(methanesulfonyl)cyclopropyl)-N-(1H-pyrazol-5-yl)pyridin-2-amine (2)

BrettPhosPd G3 (8 mg, 0.01 mmol) and Cs₂CO₃ (85 mg, 0.26 mmol) were added to a solution of (3R)-4-[6-chloro-4-(1-methanesulfonylcyclopropyl)-5-methylpyridin-2-yl] _-3 -methylmorpholine (30 mg, 0.08 mmol) and 1H-pyrazol- ₅ -amine (14 mg, 0.16 mmol) in dioxane ( _1.5 mL ₎ . The mixture was stirred for 10 h at 110 °C under a _nitrogen atmosphere. LC-MS showed that the reaction was complete. The reaction mixture was diluted with EA (40 mL), washed with water and brine, dried over anhydrous Na₂SO₄, filtered, and concentrated. The residue was purified by preparative HPLC ( _C₁₈ , 10-95% MeOH in _H₂O containing 0.1% HCOOH) to give the desired product (17 mg, yield: 49%). LC/MS(ESI):m/z 392[M+H] ⁺ . ¹ H NMR(400MHz,DMSO)δ12.09(s,1H),7.94(s,1H),7.52(s,1H),6.40(s,1H),6.24(s, 1H),4.20(d,J＝5.0Hz,1H),3.92(dd,J＝11.2,3.1Hz,1H),3.71(d,J＝11.4Hz,2H),3 .62(dd,J＝11.2,2.7Hz,1H),3.47(td,J＝11.8,2.9Hz,1H),3.06–2.98(m,1H),2.95 (s,3H),2.17(s,3H),1.87(s,1H),1.52(s,1H),1.24(s,2H),1.09(d,J=6.1Hz,3H).

Example 3

Step 1. (R)-3-methyl-N-(3-methyl-1H-pyrazol-5-yl)-6-(3-methylmorpholino)-4-(1-(methanesulfonyl)cyclopropyl)pyridine-2-amine (3)

BrettPhos-Pd-G3 (13 mg, 0.01 mmol) and Cs₂CO₃ (142 mg, 0.43 mmol) were added to a solution of (3R)-4-[6-chloro-4-(1-methanesulfonylcyclopropyl)-5-methylpyridin-2-yl]-3-methylmorpholine (50 mg, 0.14 mmol) and ₃ -methyl-1H-pyrazol- ₅ -amine (28 mg, 0.28 mmol) in dioxane (2 mL). The mixture was stirred for 10 hours at 110 °C under a _nitrogen atmosphere. LC-MS showed that the reaction was complete. The reaction mixture was diluted with EA (40 mL), washed with water and brine, dried over _{anhydrous Na₂SO₄} , filtered, and concentrated. The residue was purified by preparative HPLC ( _C18 , 10-95% MeOH in _H2O , containing 0.1% HCOOH) to obtain the desired product (54 mg, yield: 91%). LC/MS(ESI):m/z 406[M+H] ⁺ . ¹ H NMR (400MHz, DMSO) δ11.75(s,1H),7.69(s,1H),6.21(s,2H),4.20(s,1H),3.92(d,J＝8.2Hz,1H),3.71(d,J＝11.3Hz,2H),3.62(d,J＝8.8Hz,1H),3.47 (dd,J＝11.4,8.9Hz,1H),3.01(t,J＝10.9Hz,1H),2.94(s,3H),2.17(d,J＝1 5.2Hz, 6H), 1.86 (s, 1H), 1.51 (s, 1H), 1.23 (s, 2H), 1.10 (d, J = 5.9Hz, 3H).

Example 4

Step 1.2,6-Dichloro-4-(1,4-dimethyl-1H-pyrazol-5-yl)pyridine (4-3)

To a solution of 2,6-dichloro-4-iodopyridine (300 mg, 1.10 mmol) and 1,4-dimethyl-5-(tetramethyl-1,3,2-dioxacyclopentabor-2-yl)-1H-pyrazole (267.6 mg, 1.21 mmol) in DME (10 mL) _{, Na₂CO₃} (232.2 _mg , 2.19 mmol) and Pd(dppf) _Cl₂ (80.2 mg, 0.11 mmol) were added. The mixture was injected twice with _N₂ and stirred at 90 °C for 4 hours. LC-MS showed the reaction was complete. The reaction mixture was diluted with water (30 mL) and extracted with EA (40 mL × 2). The combined organic layers were washed with brine, dried over _{anhydrous Na₂SO₄} , filtered, and concentrated under vacuum. The residue was purified by preparative TLC (PE:EA = 3:1, V/V) to obtain the desired product (230 mg, yield: 86%). LC/MS (ESI) m/z: 243 [M+H] ⁺ .

Step 2. (R)-4-(6-chloro-4-(1,4-dimethyl-1H-pyrazol-5-yl)pyridin-2-yl)-3-methylmorpholine (4-5)

(3R)-3-methylmorpholine (384.4 mg, 3.80 mmol) was added to a solution of 2,6-dichloro-4-(1,4-dimethyl-1H-pyrazol-5-yl)pyridine (230 mg, 0.95 mmol) in NMP (3 mL). The reaction was stirred at 150 °C for 1 h under microwave irradiation. LC-MS showed that the reaction was complete. The mixture was diluted with water (30 mL) and extracted with EA (40 mL × 2). The combined organic layers were washed with brine, dried over _{anhydrous Na₂SO₄} , filtered, and concentrated under vacuum. The residue was purified by rapid column chromatography (silica gel, 0–10% ethyl acetate/petroleum ether) to give the desired product (150 mg, yield: 51%). LC/MS (ESI) m/z: 307 [M+H] ^⁺ .

Step 3. (R)-5-((4-(1,4-dimethyl-1H-pyrazol-5-yl)-6-(3-methylmorpholino)pyridin-2-yl)amino)-1H-pyrazol-1-carboxylic acid tert-butyl ester (4-7)

To a solution of (R)-4-(6-chloro-4-(1,4-dimethyl-1H-pyrazol-5-yl)pyridin-2-yl)-3-methylmorpholine (120 mg, 0.39 mmol) and tert-butyl 5-amino- _1H -pyrazol-1-carboxylate (107.49 mg, 0.587 mmol) in dioxane (10 mL ₎ , _CS₂CO₃ (637.2 mg, 1.96 mmol) and BrettPhos Pd G₃ (35.46 mg, 0.04 mmol) were added. The mixture was injected twice with _N₂ and stirred overnight at 90 °C. LC-MS showed the reaction was complete. The reaction mixture was diluted with water (30 mL) and extracted with EA (40 mL × 2). The combined organic layers were washed with brine, dried over anhydrous _Na₂SO₄ , filtered, and concentrated under vacuum. The residue was purified by preparative TLC (PE:EA = 2:1, V/V) to obtain the desired product (80 mg, yield: 45%). LC/MS (ESI) m/z: 454 [M+H] ⁺ .

Step 4. (R)-4-(1,4-dimethyl-1H-pyrazol-5-yl)-6-(3-methylmorpholino)-N-(1H-pyrazol-5-yl)pyridin-2-amine (4)

A mixture of (R)-5-((4-(1,4-dimethyl-1H-pyrazol-5-yl)-6-(3-methylmorpholino)pyridin-2-yl)amino)-1H-pyrazol-1-carboxylic acid tert-butyl ester (80 mg, 0.18 mmol) in HCl solution (4 M in dioxane, 2 mL) was stirred overnight at room temperature. LC-MS showed the reaction was complete. The reaction mixture was concentrated to dryness under vacuum. The residue was purified by preparative HPLC (C18, 20-95% acetonitrile in _H₂O , containing 0.1% TFA) to give the desired product (20 mg, yield: 32%). LC/MS(ESI)m/z:354[M+H] ⁺ . ¹ H NMR (400MHz, DMSO) δ9.05(s,1H),7.55(d,J＝2.2Hz,1H),7.31(s,1H),6.41(s,1H),6.35(d,J＝1.9Hz,1H),6.00(s,1H),4.30(d,J＝6.8Hz,1H),3.9 7–3.84(m,2H),3.74(s,3H),3.71(s,1H),3.63(dd,J＝11.3,2.8Hz,1H),3 .52–3.45(m,1H),3.11–3.03(m,1H),1.99(s,3H),1.15(d,J＝6.6Hz,3H).

Example 5

Step 1. (R)-2-chloro-6-(3-methylmorpholino)pyrimidine-4-carboxylic acid methyl ester (5-3)

A mixture of methyl 2,6-dichloropyrimidin-4-carboxylate (1.5 g, 7.24 mmol), (3R)-3-methylmorpholine (732 mg, 7.24 mmol), and TEA (1.47 g, 14.52 mmol) in DCM (30 mL) was stirred for 16 hours at room temperature. LC-MS showed the reaction was complete. The reaction mixture was diluted with DCM (20 mL), washed with water and brine, _dried over anhydrous _Na₂SO₄ , filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 3:1, V/V) to give the desired product (1.55 g, yield: 78%). LC/MS (ESI): m/z 272 [M+H] ^⁺ .

Step 2. (R)-(2-chloro-6-(3-methylmorpholino)pyrimidin-4-yl)methanol (5-4)

At 0 °C under a _N₂ atmosphere, _LiBH₄ solution (2.0 M in THF, 3.7 mL, 7.34 mmol) was added dropwise to a solution of methyl 2-chloro-6-[(3R)-3-methylmorpholin-4-yl]pyrimidin-4-carboxylate (1 g, 3.67 mmol) in anhydrous THF (20 mL). The resulting mixture was stirred at 0 °C for 1 hour. LC-MS showed that the reaction was complete. The reaction mixture was quenched with a saturated aqueous _NH₄Cl solution and extracted with EA (50 mL). The combined organic layers were dried over _{anhydrous Na₂SO₄} , filtered, and concentrated under vacuum. The residue was purified by silica gel column chromatography (PE:EA = 1:1, V/V) to give the desired product (800 mg, yield: 89%). LC/MS (ESI): m/z 244 [M+H] ^⁺ .

Step 3. (R)-4-(2-chloro-6-(chloromethyl)pyrimidin-4-yl)-3-methylmorpholine (5-5)

SOCl₂ (1.17 g, 9.84 mmol) was added dropwise to a solution of {2-chloro-6-[(3R)-3-methylmorpholin-4-yl]pyrimidin- ₄ -yl}methanol (800 mg, 3.28 mmol) and DMF (0.01 mL) in anhydrous DCM (20 mL) at 0 °C. The resulting mixture was stirred at room temperature for 1 hour. LC-MS showed that the reaction was complete. The reaction mixture was concentrated under reduced pressure. The residue was dissolved in EA (40 mL), washed with saturated aqueous _NaHCO₃ solution and brine, dried over _{anhydrous Na₂SO₄} , filtered, and concentrated under vacuum. The residue was used for the next step without further purification (800 mg, yield: 93%). LC/MS (ESI): m/z 262/264 [M+H] ^⁺ .

Step 4. (R)-4-(2-chloro-6-((methanesulfonyl)methyl)pyrimidin-4-yl)-3-methylmorpholine (5-6)

A mixture of (3R)-4-[2-chloro-6-(chloromethyl)pyrimidin-4-yl]-3-methylmorpholine (535 mg, 2.04 mmol) and _CH₃SO₂Na ( ₄₁₈ mg, 4.10 mmol) in DMF (10 mL) was stirred for 16 hours at room temperature. _LC -MS showed the reaction was complete. The reaction mixture was diluted with EA (40 mL), washed with water and brine, dried over anhydrous _Na₂SO₄ , filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 2:1, V/V) to give the desired product (560 mg, yield: 90%). LC/MS (ESI): m/z 306 [M+H] ^⁺ .

Step 5. (R)-4-(2-chloro-6-(1-(methanesulfonyl)cyclopropyl)pyrimidin-4-yl)-3-methylmorpholine (5-7)

A mixture of (3R)-4-[2-chloro-6-(methanesulfonylmethyl)pyrimidin-4-yl]-3-methylmorpholine (125 mg, 0.41 mmol), 1,2-dibromoethane (154 mg, 0.82 mmol), NaOH (10.0 M in _H₂O , 0.4 mL, 4.0 mmol), and TBAB (26 mg, 0.08 mmol) in toluene (4 mL) was stirred for 3 hours at 60 °C. _LC -MS showed the reaction was complete. The reaction mixture was diluted with EA (40 mL), washed with water and brine, dried over anhydrous _Na₂SO₄ , filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 1:1, V/V) to give the desired product (110 mg, yield: 81%). LC/MS (ESI): m/z 332 [M+H] ^⁺ .

Step 6. (R)-5-((4-(3-methylmorpholino)-6-(1-(methanesulfonyl)cyclopropyl)pyrimidin-2-yl)amino)-1H-pyrazole-1-carboxylic acid tert-butyl ester (5-9)

Pd₂(dba)₃ (55 mg, 0.06 mmol), Xant-Phos (34 mg, 0.06 mmol), and Cs₂CO₃ (394 mg, 1.21 mmol) were added to a solution of (3R) _-4- [2-chloro-6-(1-methanesulfonylcyclopropyl)pyrimidin-4-yl] _-3- methylmorpholine (200 mg, 0.60 mmol) and tert-butyl 5-amino-1H-pyrazole- _{1 -} carboxylate (166 mg, 0.90 mmol) in dioxane (10 mL). The mixture was stirred for 6 hours at 100 °C under a _nitrogen atmosphere. LC-MS showed that the reaction was complete. The reaction mixture was diluted with EA (40 mL), washed with water and brine, _dried over anhydrous _Na₂SO₄ , filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 1:2, V/V) to obtain the desired product (129 mg, yield: 44%). LC/MS (ESI): m/z 479 [M+H] ⁺ .

Step 7. (R)-4-(3-methylmorpholino)-6-(1-(methanesulfonyl)cyclopropyl)-N-(1H-pyrazol-5-yl)pyrimidin-2-amine (5)

A mixture of 5-{[4-(1-methanesulfonylcyclopropyl)-6-[(3R)-3-methylmorpholin-4-yl]pyrimidin-2-yl]amino}-1H-pyrazole-1-carboxylic acid tert-butyl ester (60 mg, 0.12 mmol) in HCl solution (4.0 M in dioxane, 3.0 mL) was stirred for 10 hours at room temperature. LC-MS showed the reaction was complete. The reaction mixture was concentrated under reduced pressure. The residue was purified by preparative HPLC ( _C18 , 10-95% MeOH in _H2O containing 0.1% HCOOH) to give the desired product (20 mg, yield: 42%). LC/MS(ESI):m/z 379[M+H] ⁺ . ¹ H NMR (400MHz, DMSO) δ12.29(s,1H),9.51(s,1H),7.58(s,1H),6.38(s,2H),4.44(s,1H),4.05(d,J＝12.8Hz,1H),3.94(dd,J＝11.4,3.4Hz,1H),3.74 (d,J＝11.4Hz,1H),3.59(dd,J＝11.5,2.9Hz,1H),3.46(s,1H),3.25(s,3H ), 3.18 (s, 1H), 1.60 (t, J = 5.7Hz, 2H), 1.50 (s, 2H), 1.21 (d, J = 6.7Hz, 3H).

Example 6

Step 1.5-{[4-(1-methanesulfonylcyclopropyl)-6-[(3R)-3-methylmorpholin-4-yl]pyrimidin-2-yl]amino}-3-methyl-1H-pyrazole-1-carboxylic acid tert-butyl ester (6-2)

Cs₂CO₃ (196.4 mg, 0.60 mmol), Xant-Phos (17.4 mg, 0.03 mmol), and Pd₂(dba)₃ (24.4 mg, 0.03 mmol) were added to a solution of (3R)-4-[2-chloro-6-(1-methanesulfonylcyclopropyl)pyrimidin- ₄ -yl] _-3- methylmorpholine (100 mg, 0.30 mmol) and 5-amino- _3- methyl-1H-pyrazole- _1- carboxylic acid tert-butyl ester (89.2 mg, 0.45 mmol) in dioxane (5 mL). The mixture was stirred for 6 hours at 100 °C under a nitrogen atmosphere.

LC-MS showed the reaction was complete. The reaction mixture was diluted with EA (40 mL), washed with water and brine, _dried over anhydrous _Na₂SO₄ , filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 1:1, V/V) to give the desired product (130 mg, yield: 87%). LC/MS (ESI): m/z 493 [M+H] ^⁺ .

Step 2.4-(1-Methanesulfonylcyclopropyl)-N-(3-methyl-1H-pyrazol-5-yl)-6-[(3R)-3-methylmorpholin-4-yl]pyrimidin-2-amine (6)

To a solution of 5-{[4-(1-methanesulfonylcyclopropyl)-6-[(3R)-3-methylmorpholin-4-yl]pyrimidin-2-yl]amino}-3-methyl-1H-pyrazole-1-carboxylic acid tert-butyl ester (120 mg, 0.24 mmol) in DCM (2 mL), HCl solution (4 M in dioxane, 2 mL) was added. The mixture was stirred at room temperature for 2 hours. LC-MS showed that the reaction was complete. The reaction mixture was concentrated under vacuum. The residue was purified by preparative HPLC (C18, 10-95% MeCN in _H₂O , containing 0.1% ammonia) to give the desired product (32.6 mg, yield: 34%). LC/MS(ESI)m/z:393[M+H] ⁺ . ¹ H NMR (400MHz, DMSO) δ9.21(s,1H),6.31(s,1H),6.15(s,1H),4.40(s,1H),4.02(d,J＝11.7Hz,1H),3.93(d,J＝8.1Hz,1H),3.73(d,J＝11.3H z,1H),3.58(dd,J＝11.6,2.9Hz,2H),3.25(s,3H),3.16(d,J＝10.8Hz,1H),2.19(s,3H),1.58(s,2H),1.47(s,2H),1.20(d,J＝6.7Hz,3H).

Compound 6 can be prepared using the following method:

Step 1. 3-Amino-5-methyl-1H-pyrazole-1-carboxylic acid tert-butyl ester (6-3)

At 0 °C, NaH (60%, 10.81 g, 270.28 mmol) was added in portions to a solution of 3-methyl-1H-pyrazole-5-amine (25 g, 257.41 mmol) in THF (800 mL). After stirring at 0 °C for 30 min, (Boc) _₂O (58.99 g, 270.28 mmol) was added in a single batch. The mixture was stirred at _room temperature for 1 h. TLC showed that the reaction was complete. The reaction mixture was poured into a saturated aqueous solution of _NH₄Cl and extracted twice with DCM (600 mL × 2). The combined organic layers were separated, washed with brine, dried over anhydrous _Na₂SO₄ , filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 2:1, V/V) to give the desired product (19 g, yield: 37.42%). ¹ HNMR (400MHz, CDCl ₃ ) δ 5.59 (d, J = 0.9 Hz, 1H), 3.89 (s, 2H), 2.44 (d, J = 0.9 Hz, 3H), 1.62 (s, 9H).

Step 2. (R)-5-methyl-3-((4-(3-methylmorpholino)-6-(1-(methanesulfonyl)cyclopropyl)pyrimidin-2-yl)amino)-1H-pyrazole-1-carboxylic acid tert-butyl ester (6-4)

BrettPhos-Pd-G3 (906 mg, 4.41 mmol) and Cs₂CO₃ (29.45 g, 90.4 mmol) were added to a solution of (3R)-4-[2-chloro-6-(1-methanesulfonylcyclopropyl)pyrimidin-4-yl]-3-methylmorpholine (15.0 g, 45.20 mmol) and tert-butyl ₃ -amino- _{5-methyl-1H-pyrazole-1} -carboxylate (10.7 g, 54.24 _mmol ) in dioxane (600 mL). The mixture was stirred overnight at 100 °C under a _nitrogen atmosphere. The reaction mixture was diluted with EA (1.0 L), washed with water and brine, dried over anhydrous _Na₂SO₄ , filtered, and concentrated. The residue was purified by silica gel column chromatography (DCM:MeOH = 20:1, V/V) to give the desired product (17 g, yield: 76%). LC/MS(ESI): m/z 493[M+H] ⁺ .

Step 3. (R)-N-(3-methyl-1H-pyrazol-5-yl)-4-(3-methylmorpholino)-6-(1-(methanesulfonyl)cyclopropyl)pyrimidin-2-amine (6)

A mixture of (R)-5-methyl-3-((4-(3-methylmorpholino)-6-(1-(methanesulfonyl)cyclopropyl)pyrimidin-2-yl)amino)-1H-pyrazole-1-carboxylic acid tert-butyl ester (17.0 g, _34.51 mmol) in HCl solution (4.0 M in dioxane, 100.0 mL) was stirred for 12 hours at room temperature. The reaction mixture was concentrated to dryness under reduced pressure, and the residue was diluted with EA (200 mL) and saturated NaHCO3 aqueous solution (200 mL). The resulting mixture was stirred overnight at room temperature. The organic layer was separated, washed with brine, dried over _{anhydrous Na2SO4} , filtered, and concentrated. The residue was purified by preparative HPLC ( _C18 , 10-95% MeOH in _H2O containing 0.1% HCOOH) to give the desired product (10 g, yield: 73%). LC/MS(ESI):m/z 393[M+H] ⁺ . ¹ H NMR (400MHz, DMSO) δ11.78(s,1H),9.10(s,1H),6.23(d,J＝29.9Hz,2H),4.38(s,1H),4.07–3.87(m,2H),3.73(d,J＝11.4Hz,1H),3.58 (dd,J＝11.5,2.9Hz,1H),3.43(td,J＝11.8,2.9Hz,1H),3.26(s,3H),3.13(td,J＝12.9,3.7Hz,1H),2.19(s,3H),1.19(d,J＝6.7Hz,3H).

Example 7

Step 1. (R)-4-(2-chloro-6-(2-(methanesulfonyl)prop-2-yl)pyrimidin-4-yl)-3-methylmorpholine (7-1)

At 0 °C, a solution of CH₃I (1.26 g, 8.85 mmol) in anhydrous DMF (1 mL) was added dropwise to a solution of (3R)-4-[ ₂ -chloro-6-(methanesulfonylmethyl)pyrimidin-4-yl]-3-methylmorpholine (900 mg, 2.94 mmol) and t-BuONa (849 mg, 8.82 mmol) in anhydrous DMF (16 mL). After addition, the resulting mixture was stirred at room temperature for 3 hours. _LC -MS showed that the reaction was complete. The reaction mixture was diluted with EA (40 mL), washed with water and brine, dried over anhydrous _Na₂SO₄ , filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 10:1, V/V) to give the desired product (870 mg, yield: 88%). LC/MS (ESI): m/z 334 [M+H] ^⁺ .

Step 2. (R)-4-(3-methylmorpholino)-6-(2-(methanesulfonyl)propyl-2-yl)-N-(1H-pyrazol-5-yl)pyrimidin-2-amine (7)

A mixture of (3R)-4-[2-chloro- _6- (2-methanesulfonylprop-2-yl)pyrimidin-4-yl]-3-methylmorpholine (100 mg, 0.30 mmol), 1H-pyrazol-5-amine (37 mg, 0.44 mmol), BrettPhos Pd G3 (27 mg, 0.03 mmol), and _Cs₂CO₃ (293 mg, 0.90 mmol) in _dioxane (5 mL) was stirred for 10 hours at 110 °C under a N₂ atmosphere. LC-MS showed the reaction was complete. The reaction mixture was diluted with EA (40 mL), washed with water and brine, dried over _{anhydrous Na₂SO₄} , filtered, and concentrated under vacuum. The residue was purified by preparative HPLC ( _C₁₈ , 10-95% MeOH in _H₂O containing 0.1% HCOOH) to give the desired product (36.7 mg, yield: 32%). LC/MS(ESI):m/z 381[M+H] ⁺ . ¹ H NMR(400MHz,DMSO)δ12.18(s,1H),9.26(s,1H),7.52(s,1H),6.41(s,1H),6.30 (s,1H),4.42(s,1H),4.03(d,J＝12.9Hz,1H),3.94(dd,J＝11.4,3.3Hz,1H),3.7 3(d,J＝11.4Hz,1H),3.59(dd,J＝11.5,3.0Hz,1H),3.44(dd,J＝11.8,9.0Hz,1H) ,3.14(td,J＝12.9,3.7Hz,1H),3.01(s,3H),1.67(s,6H),1.19(d,J＝6.7Hz,3H).

Example 8

Step 1. (R)-N-(3-methyl-1H-pyrazol-5-yl)-4-(3-methylmorpholino)-6-(2-(methanesulfonyl)propyl-2-yl)pyrimidin-2-amine (8)

A mixture of (3R)-4-[6-chloro- _4- (2-methanesulfonylprop-2-yl)pyridin-2-yl]-3-methylmorpholine (100 mg, 0.30 mmol), 3-methyl-1H-pyrazol-5-amine (58 mg, 0.60 mmol), BrettPhos Pd G3 (27 mg, 0.03 mmol), and _Cs₂CO₃ (293 _mg , 0.90 mmol) in dioxane (4 mL) was stirred for 10 hours at 110 °C under a N₂ atmosphere. LC-MS showed that the reaction was complete. The reaction mixture was diluted with EA (40 mL), washed with water and brine, dried over _{anhydrous Na₂SO₄} , filtered, and concentrated under vacuum. The residue was purified by preparative HPLC ( _C18 , 10-95% MeOH in _H2O , containing 0.1% HCOOH) to obtain the desired product as a white solid (44.8 mg, yield: 37%). LC/MS (ESI): m/z 395 [M+H] ⁺ . ¹ HNMR (400MHz, DMSO) δ 11.98 (s, 1H), 9.08 (s, 1H), 8.13 (s, 1H), 6.28 (s, 1H), 6.16 (s, 1H), 4.40 (s, 1H), 4.02 (d, J = 13.0 Hz, 1H), 3.93 (dd, J = 11.4, 3.4 Hz, 1H), 3.73 (d, J ＝11.4Hz,1H),3.59(dd,J＝11.5,2.9Hz,1H),3.44(td,J＝11.8,2.8Hz,1H),3.13(td ,J＝13.0,3.7Hz,1H),3.01(s,3H),2.18(s,3H),1.66(s,6H),1.19(d,J＝6.7Hz,3H).

Example 9

Step 1. (R)-(2-chloro-6-(3-methylmorpholino)pyrimidin-4-yl)methanesulfonate (9-1)

At 0 °C, MsCl (564 mg, 4.92 mmol) in DCM (2 mL) was added dropwise to a solution of (R)-(2-chloro-6-(3-methylmorpholino)pyrimidin-4-yl)methanol (1 g, 4.10 mmol) and TEA (623 mg, 6.15 mmol) in DCM (30 mL). The resulting mixture was stirred at room temperature for 3 hours. _LC -MS showed that the reaction was complete. The reaction mixture was diluted with EA (40 mL), washed with water and brine, dried over anhydrous _Na₂SO₄ , filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 10:1, V/V) to give the desired product (1.06 mg, yield: 80%). LC/MS (ESI): m/z 322 [M+H] ^⁺ .

Step 2. (R)-2-(2-chloro-6-(3-methylmorpholino)pyrimidin-4-yl)acetonitrile (9-2)

A solution of (R)-(2-chloro-6-(3-methylmorpholino)pyrimidin-4-yl)methanesulfonate (1 g, 3.10 mmol) was added dropwise to a solution of NaCN (184 mg, 3.75 mmol) in DMSO (20 mL). The resulting mixture was stirred at room temperature for 1 hour. LC-MS showed that the reaction was complete. The reaction mixture was diluted with EA (40 mL), washed with ice water and brine, dried over anhydrous _{Na₂SO₄ ,} filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 3:1, V/V) to give the desired product (300 mg, yield: 38%). LC/MS (ESI): m/z 253 [M+H] ^⁺ .

Step 3. (R)-1-(2-chloro-6-(3-methylmorpholino)pyrimidin-4-yl)cyclopropaneformonitrile (9-3)

A solution of KOH (1.57 g, 28.0 mmol) in H₂O (15 mL) was added to a solution of (R)-2-(2-chloro-6-(3-methylmorpholino)pyrimidin-4-yl)acetonitrile (100 mg, 0.40 mmol), 1,2-dibromoethane (338 mg, 1.79 mmol), and TBAB (32.2 mg, 0.1 mmol) in ₂ -MeTHF (15 mL). The resulting mixture was stirred at room temperature for 12 hours. LC-MS showed that the reaction was complete. The reaction mixture was diluted with EA (50 mL), washed with water and brine, _dried over anhydrous _Na₂SO₄ , filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 5:1, V/V) to give the desired product (50 mg, yield: 46%). LC/MS (ESI): m/z 279 [M+H] ^⁺ .

Step 4. (R)-1-(2-((3-methyl-1H-pyrazol-5-yl)amino)-6-(3-methylmorpholino)pyrimidin-4-yl)cyclopropaneformonitrile (9)

BrettPhos Pd _G3 (16 mg, 0.018 mmol) was added to a solution of (R)-1-(2-chloro-6-(3-methylmorpholino)pyrimidin-4-yl)cyclopropaneformitrile (50 mg, 0.18 mmol), 3-methyl-1H-pyrazol _-5- amine (35 mg, 0.36 mmol), and Cs₂CO₃ (117.3 mg, 0.36 mmol) in dioxane (5 mL). The mixture was stirred at 100 °C for 16 h. LC-MS showed that the reaction was complete. The reaction mixture was diluted with EA (40 mL), washed with water and brine, dried over _{anhydrous Na₂SO₄} , filtered, and concentrated under vacuum. The residue was purified by preparative HPLC ( _C₁₈ , 10-95% MeOH in _H₂O containing 0.1% HCOOH) to give the desired product (8.2 mg, yield: 13%). LC/MS(ESI):m/z 340[M+H] ⁺ . ¹ H NMR (400MHz, DMSO) δ8.97(s,1H),6.17(s,2H),4.35(s,1H),4.04–3.87(m,2H),3.72(d,J＝11.4Hz,1H),3.58(dd,J＝ 11.5,2.9Hz,1H),3.47–3.39(m,4H),3.13(td,J＝12.9,3.7Hz,2H),2.17(s,3H),1.70(s,4H),1.19(d,J＝6.7Hz,3H).

Example 10

Step 1. (R)-2-(2-chloro-6-(3-methylmorpholino)pyrimidin-4-yl)-2-methylpropionitrile (10-1)

At 0 °C, a solution of CH₃I (605 mg, 4.26 mmol) in anhydrous THF (1 mL) was added dropwise to a solution of (R)-( ₂ -chloro-6-(3-methylmorpholino)pyrimidin-4-yl)methanesulfonate (360 mg, 1.42 mmol) and t-BuONa (274 mg, 2.85 mmol) in anhydrous THF (15 mL). After addition, the resulting mixture was stirred at room temperature for 12 hours. _LC -MS showed that the reaction was complete. The reaction mixture was diluted with EA (40 mL), washed with water and brine, dried over anhydrous _Na₂SO₄ , filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 5:1, V/V) to give the desired product (300 mg, yield: 75%). LC/MS (ESI): m/z 281 [M+H] ^⁺ .

Step 2. (R)-2-methyl-2-(2-((3-methyl-1H-pyrazol-5-yl)amino)-6-(3-methylmorpholino)pyrimidin-4-yl)propionitrile (10)

BrettPhos Pd G3 (16 mg, 0.018 mmol) was added to a solution of (R)-2-(2-chloro-6-(3-methylmorpholino)pyrimidin-4-yl)-2-methylpropionitrile (50 mg, 0.18 mmol), tert-butyl 5-amino-3-methyl-1H-pyrazole- _{1 -} carboxylate (70 mg, 0.36 mmol), and Cs₂CO₃ (174 mg, 0.53 mmol) in dioxane (5 _mL ). The mixture was stirred at 100 °C for 16 h. LC-MS showed that the reaction was complete. The reaction mixture was diluted with EA (40 mL), washed with water and brine, dried over anhydrous _Na₂SO₄ , filtered, and concentrated under vacuum. The residue was dissolved in DCM (4 mL), and then HCl solution (4 M in dioxane, 2 mL) was added. The mixture was stirred at room temperature for 2 h. LC-MS showed that the reaction was complete. The reaction mixture was concentrated under vacuum. The residue was purified by preparative HPLC ( _C18 , 10-95% MeOH in _H2O , containing 0.1% HCOOH) to obtain the desired product (8 mg, yield: 13%). LC/MS(ESI):m/z 342[M+H] ⁺ . ¹ H NMR (400MHz, DMSO) δ11.78(s,1H),9.07(s,1H),6.26(d,J＝17.8Hz,2H),4.40(dd,J＝13.5,7.2Hz,1H),4.01(d,J＝13.2Hz,1H),3.93(dd,J＝11.3,3.3H z,1H),3.72(d,J＝11.4Hz,1H),3.58(dd,J＝11.4,2.9Hz,1H),3.45–3.42(m ,1H),3.17–3.10(m,1H),2.17(s,3H),1.64(s,6H),1.19(d,J＝6.7Hz,3H).

Example 11

Step 1. (R)-4-(2-chloro-6-(4-(methanesulfonyl)tetrahydro-2H-pyran-4-yl)pyrimidin-4-yl)-3-methylmorpholine (11-1)

A mixture of (3R)-4-[2-chloro-6-(methanesulfonylmethyl)pyrimidin-4-yl]-3-methylmorpholine (400 mg, 1.31 mmol), 1-bromo-2-(2-bromoethoxy)ethane (905 mg, 3.93 mmol), TBAB (42 mg, 0.13 mmol), and NaOH (10.0 M in _H₂O , 1.31 mL, 13.1 mmol) in DCM (20 mL) was stirred for 24 hours at room temperature. LC-MS showed the reaction was complete. The reaction mixture was diluted with DCM (40 mL), washed with water and brine, dried over _{anhydrous Na₂SO₄} , filtered, and concentrated under vacuum. The residue was purified by silica gel column chromatography (PE:EA = 3:1, V/V) to give the desired product (147 mg, yield: 30%). LC/MS (ESI): m/z 376 [M+H] ^⁺ .

Step 2. (R)-4-(3-methylmorpholino)-6-(4-(methanesulfonyl)tetrahydro-2H-pyran-4-yl)-N-(1H-pyrazol-5-yl)pyrimidin-2-amine (11)

A mixture of (3R)-4-[2-chloro- _6- (4-methanesulfonyltetrahydropyran-4-yl)pyrimidin-4-yl]-3-methylmorpholine (70 mg, 0.19 mmol), 1H-pyrazol-5-amine (31 mg, 0.37 mmol), BrettPhos Pd G3 (17 mg, 0.02 mmol), and _Cs₂CO₃ (182 mg, 0.56 mmol) in _dioxane (3 mL) was stirred for 10 hours at 110 °C under a N₂ atmosphere. LC-MS showed the reaction was complete. The reaction mixture was diluted with EA (40 mL), washed with water and brine, dried over _{anhydrous Na₂SO₄} , filtered, and concentrated under vacuum. The residue was purified by preparative HPLC ( _C₁₈ , 10–95% MeOH in _H₂O containing 0.1% HCOOH) to give the desired product (40 mg, yield: 50%). LC/MS(ESI):m/z 423[M+H] ⁺ . ¹ H NMR (400MHz, DMSO) δ12.09(s,1H),9.19(s,1H),7.53(s,1H),6.40(s,2H),4.41(d,J＝4.6H z,1H),4.08(d,J＝12.8Hz,1H),3.91(ddd,J＝16.0,10.8,4.0Hz,3H),3.73(d,J＝11.5Hz,1H) ,3.61(dd,J＝11.5,2.9Hz,1H),3.46(td,J＝11.9,2.9Hz,1H),3.17(ddd,J＝19.1,16.3,8.1H z, 3H), 2.85 (s, 3H), 2.64 (d, J = 13.1Hz, 2H), 2.13 (t, J = 11.8Hz, 2H), 1.19 (d, J = 6.7Hz, 3H).

Example 12

Step 1. (R)-N-(3-methyl-1H-pyrazol-5-yl)-4-(3-methylmorpholino)-6-(4-(methanesulfonyl)tetrahydro-2H-pyran-4-yl)pyrimidin-2-amine (12)

A mixture of (3R)-4-[2-chloro- _6- (4-methanesulfonyltetrahydropyran-4-yl)pyrimidin-4-yl]-3-methylmorpholine (70 mg, 0.18 mmol), 3-methyl-1H-pyrazol-5-amine (36 mg, 0.37 mmol), BrettPhosPd G3 (17 mg, 0.02 mmol), and _Cs₂CO₃ (182 _mg , 0.56 mmol) in dioxane (3 mL) was stirred for 10 hours at 110 °C under a _N₂ atmosphere. LC-MS showed that the reaction was complete. The reaction mixture was diluted with EA (40 mL), washed with water and brine, dried over anhydrous _Na₂SO₄ , filtered, and concentrated under vacuum. The residue was purified by preparative HPLC ( _C18 , 10-95% MeOH in _H2O , containing 0.1% HCOOH) to obtain the desired product (48 mg, yield: 61%). LC/MS (ESI): m/z 437 [M+H] ⁺ . ^1H NMR (400MHz, DMSO) δ 11.78 (s, 1H), 9.03 (s, 1H), 8.16 (s, 1H), 6.38 (s, 1H), 6.16 (s, 1H), 4.39 (s, 1H), 4.08 (d, J = 13.0 Hz, 1H), 3.98–3.84 (m, 3H), 3.72 (d, J = 11.4 Hz, 1H), 3.6 1(dd,J＝11.4,2.9Hz,1H),3.46(td,J＝11.9,2.8Hz,1H),3.16(ddd,J＝19.2,16.3,8.1H z, 3H), 2.85 (s, 3H), 2.63 (d, J = 13.0Hz, 2H), 2.24–2.02 (m, 5H), 1.19 (d, J = 6.7Hz, 3H).

Example 13

Step 1. (R)-N-(2-chloro-6-(3-methylmorpholino)pyridin-4-yl)methanesulfonamide (13-2)

(3R)-3-methylmorpholine (629 mg, 6.22 mmol) was added to a solution of N-(2,6-dichloropyridin-4-yl)methanesulfonamide (500 mg, 2.07 mmol) in NMP (15 mL). The mixture was stirred for 1 hour at 170 °C under microwave irradiation. _LC -MS showed that the reaction was complete. The mixture was diluted with water (60 mL) and extracted three times with EA (30 mL × 3). The combined organic phases were washed with brine, dried over _Na₂SO₄ , filtered, and concentrated to dryness. The residue was purified by silica gel column chromatography (PE:EA = 1:1, V/V) to give the desired product (425 mg, yield: 67.02%). LC/MS (ESI): m/z 306 [M+H] ^⁺ .

Step 2. (R)-N-(2-chloro-6-(3-methylmorpholino)pyridin-4-yl)-N-methylmethanesulfonamide (13-3)

MeI (174 mg, 1.23 mmol) was added to a mixture of (R)-N-(2-chloro-6-(3-methylmorpholino)pyridin- ₄ -yl)methanesulfonamide (250 mg, 0.82 mmol) and _K₂CO₃ (339 mg, 2.45 mmol) in DMF (8 mL). The mixture was stirred at room temperature for 2 hours. LC-MS showed that the reaction was complete. The reaction mixture was poured into _H₂O (30 mL) and extracted three times with EA (30 mL × 3). The combined organic phases were washed with brine, dried over _{anhydrous Na₂SO₄} , filtered, and concentrated to dryness. The residue was purified by silica gel column chromatography (PE:EA = 1:1, V/V) to give the desired product (218 mg, yield: 83.4%). LC/MS (ESI): m/z 320 [M+H] ^⁺ .

Step 3. (R)-N-methyl-N-(2-(3-methylmorpholino)-6-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-5-yl)amino)pyridin-4-yl)methanesulfonamide (13-5)

Pd₂(dba)₃ (29 mg, 0.031 mmol) and XantPhos (36 mg, 0.06 mmol) were added to a solution of (R)-N-(2-chloro-6-(3-methylmorpholino)pyridin-4-yl)-N-methylmethanesulfonamide (100 mg, 0.31 mmol), 1-({[2-(trimethylsilyl)ethoxy]methyl}-λ^2-chloroalyl) _-1H -pyrazol- _{5- amine (} 100 mg, 0.47 mmol) and Cs₂CO₃ (306 mg, 0.94 mmol) in dioxane (5 mL). The mixture was stirred for 16 hours at 100 °C under a _nitrogen atmosphere. LC-MS showed that the reaction was complete. The reaction mixture was diluted with EA (60 mL), washed with water and brine, dried over anhydrous _{Na₂SO₄ ,} filtered, and concentrated to dryness. The residue was purified by silica gel column chromatography (PE:EA = 1:1, V/V) to obtain the desired product (118 mg, yield: 76%). LC/MS (ESI): m/z 497 [M+H] ⁺ .

Step 4. (R)-N-(2-((1H-pyrazol-5-yl)amino)-6-(3-methylmorpholino)pyridin-4-yl)-N-methylmethanesulfonamide (13)

A mixture of (R)-N-methyl-N-(2-(3-methylmorpholino)-6-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-5-yl)amino)pyridin-4-yl)methanesulfonamide (118 mg, 0.24 mmol) in TBAF solution (1 M in THF, 2 mL) was stirred for 2 hours at 60 °C. _LC -MS showed the reaction was complete. The mixture was diluted with _H₂O and extracted three times with EA (30 mL × 3). The combined organic phases were washed with brine, dried over anhydrous _Na₂SO₄ , filtered, and concentrated to dryness. The residue was purified by preparative HPLC (C ₁₈ , 10-95% MeOH in _H₂O containing 0.1% HCOOH) to give the desired product (20 mg, yield: 23%). LC/MS(ESI):m/z 367[M+H] ⁺ . ¹ H NMR (400MHz, DMSO) δ8.96(s,1H),7.53(d,J＝2.2Hz,1H),6.47(s,1H),6.30(d,J＝2.1Hz,1H),6.05(d,J＝1.2Hz,1H),4.29–4.23(m,1H),3.93(dd,J＝11. 2,3.2Hz,1H),3.77–3.71(m,2H),3.64–3.61(m,1H),3.50–3.49(m,1H),3.1 9(s,3H),3.07(dd,J=12.6,3.7Hz,1H),3.01(s,3H),1.13(d,J=6.6Hz,3H).

Example 14

Step 1. (R)-6-(3-methylmorpholino)-4-(1-(methanesulfonyl)cyclopropyl)-N-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-5-yl)pyridine-2-amine (14-1)

Pd₂(dba)₃ (98.5 mg, 0.11 mmol), BrettPhos-Pd-G₃ (13.7 mg, 0.015 mmol), and Cs₂CO₃ (701.2 mg, 2.15 mmol) were added to a solution of (R)-4-(6-chloro-4-(1-(methanesulfonyl)cyclopropyl)pyridin-2-yl)-3-methylmorpholine (356.0 mg, 1.08 mmol) and ₁ -({[ _2- (trimethylsilyl)ethoxy]methyl}-λ²⁻-chloroalyl)-1H-pyrazol- ₅ -amine (343.8 mg, 1.61 mmol) in _dioxane (15 mL). The mixture was stirred overnight at 100 °C under a nitrogen atmosphere. LC-MS showed that the reaction was complete. The reaction mixture was diluted with EA (40 mL), washed with water and brine, dried over _{anhydrous Na₂SO₄} , filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 3:1, V/V) to give the desired product (490 mg, yield: 89%). LC/MS (ESI) m/z: 508 [M+H] ^⁺ .

Step 2. (R)-N-methyl-6-(3-methylmorpholino)-4-(1-(methanesulfonyl)cyclopropyl)-N-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-5-yl)pyridine-2-amine (14-2)

At 0 °C, NaH (60%, 14.2 mg, 0.59 mmol) was added dropwise to a solution of (R)-6-(3-methylmorpholino)-4-(1-(methanesulfonyl)cyclopropyl)-N-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-5-yl)pyridine-2-amine (200 mg, 0.39 mmol) in THF (5 mL). The mixture was stirred at 0 °C for 30 min, and then _CH3I (84.0 mg, 0.59 mmol) was added dropwise to a solution of THF (1 mL). The resulting mixture was stirred at room temperature for another 1 h. _LC -MS showed that the reaction was complete. The reaction mixture was quenched with saturated _NH4Cl aqueous solution and extracted with EA (30 mL × 2). The combined organic layers were washed with brine, dried over anhydrous _Na2SO4 , filtered, and concentrated. The residue was purified by silica gel column chromatography (DCM:MeOH = 50:1, V/V) to obtain the desired product (110 mg, yield: 53%). LC/MS (ESI) (m/z): 522 [M+H] ⁺ .

Step 3. (R)-N-methyl-6-(3-methylmorpholino)-4-(1-(methanesulfonyl)cyclopropyl)-N-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-5-yl)pyridine-2-amine (14)

A mixture of (R)-N-methyl-6-(3-methylmorpholino)-4-(1-(methanesulfonyl)cyclopropyl)-N-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-5-yl)pyridine-2-amine (110 mg, 0.21 mmol) in HCl solution (4 M in dioxane, 2 mL) was stirred for 2 hours at room temperature. LC-MS showed the reaction was complete. The reaction mixture was concentrated under vacuum. The residue was purified by preparative HPLC (C18, 10-95% MeCN in _H₂O containing 0.1% HCOOH) to give the desired product (14 mg, yield: 17%). LC/MS (ESI) m/z: 392 [M+H] ⁺ . ¹ H NMR(400MHz,DMSO)δ12.40(s,1H),8.37(s,1H),7.66(s,1H),6.48(s,1H),6.24(s,1H ),6.21(d,J＝1.9Hz,1H),4.27(d,J＝4.9Hz,1H),3.97–3.80(m,3H),3.72(d,J＝11.2Hz, 1H),3.62(dd,J＝11.3,2.8Hz,2H),3.53–3.42(m,3H),3.05(td,J＝12.6,3.6Hz,2H),2 .91 (s, 4H), 1.53 (dd, J = 6.2, 4.1Hz, 3H), 1.22 (t, J = 5.1Hz, 3H), 1.14 (d, J = 6.6Hz, 4H).

Example 15

Step 1.2, 6-Dichloro-4-iodonicotinaldehyde (15-2)

At -78°C, LDA solution (2M in THF, 2.74 mL, 5.48 mmol) was added dropwise to a solution of 2,6-dichloro-4-iodopyridine (1 g, 3.65 mmol) in THF (10 mL). The mixture was stirred at -78°C for 1 hour, followed by dropwise addition of THF (1 mL) containing ethyl formate (0.44 mL, 5.48 mmol). The resulting mixture was stirred at -78°C for another 2 hours. LC-MS showed the reaction was complete. The reaction mixture was quenched with saturated _NH4Cl aqueous solution and extracted twice with EA (40 mL × 2). The combined organic layers were washed with brine, dried over _{anhydrous Na2SO4} , filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 30:1, V/V) to give the desired product (553 mg, yield: 50%). LC/MS (ESI): m/z 302[M+H] ⁺ .

Step 2.2,6-Dichloro-4-(1,4-dimethyl-1H-pyrazol-5-yl)nicotinaldehyde (15-3)

Pd(dppf)Cl₂ (360 mg, 0.50 mmol) and Na₂CO₃ (2.0 M in H₂O, 6 mL, 12.0 mmol) were added to a solution of 2,6-dichloro-4-iodopyridin-3-carboxaldehyde (1.5 g, 4.97 mmol) and _1,4- dimethyl- _5- (tetramethyl-1,3,2-dioxanepentobor- _{2 -} yl)-1H-pyrazole (1.32 g, 5.96 mmol) in DME (90 mL). The mixture was injected twice with _N₂ and stirred overnight at 100 °C under a _N₂ atmosphere. LC-MS showed the reaction was complete. The reaction mixture was diluted with _H₂O (100 mL) and extracted twice with EA (100 mL × 2). The combined organic layers were washed with brine, _dried over anhydrous _Na₂SO₄ , filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 10:1, V/V) to obtain the desired product (548 mg, yield: 41%). LC/MS (ESI): m/z 270 [M+H] ⁺ . ¹ H NMR (400MHz, DMSO) δ 9.98 (s, 1H), 7.79 (s, 1H), 7.37 (s, 1H), 3.59 (s, 3H), 1.79 (s, 3H).

Step 3. (R)-2-chloro-4-(1,4-dimethyl-1H-pyrazol-5-yl)-6-(3-methylmorpholino)nicotinaldehyde (15-4)

A solution of 2,6-dichloro-4-(1,4-dimethyl-1H-pyrazol-5-yl)pyridine-3-carboxaldehyde (330 mg, 1.22 mmol) and (3R)-3-methylmorpholine (185 mg, 1.83 mmol) in NMP (14 mL) was stirred for 1 hour at 130 °C under microwave irradiation. LC-MS showed the reaction was complete. The reaction mixture was quenched with _H₂O (40 mL) and extracted twice with EA (50 mL × 2). The combined organic layers were washed with brine, _dried over anhydrous _Na₂SO₄ , filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 5:1, V/V) to give the desired product (142 mg, yield: 35%). LC/MS (ESI): m/z 335 [M+H] ^⁺ .

Step 4. (R)-(2-chloro-4-(1,4-dimethyl-1H-pyrazol-5-yl)-6-(3-methylmorpholino)pyridin-3-yl)methanol(15-5)

To a solution of 2-chloro-4-(1,4-dimethyl-1H-pyrazol-5-yl)-6-[(3R)-3-methylmorpholin-4-yl]pyridine-3-carboxaldehyde (140 mg, 0.42 mmol) in THF ( ₄ mL), NaBH₄ (14 mg, 0.42 mmol) was added. The resulting mixture was stirred at 0 °C for 0.5 h. LC-MS showed that the reaction was complete. The reaction mixture was quenched with _H₂O and extracted twice with EA (40 mL × 2). The combined organic layers were washed with brine, dried over _{anhydrous Na₂SO₄} , filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 3:1, V/V) to give the desired product (140 mg, yield: 99%). LC/MS (ESI): m/z 337 [M+H] ^⁺ .

Step 5. (R)-(4-(1,4-dimethyl-1H-pyrazol-5-yl)-6-(3-methylmorpholino)-2-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-5-yl)amino)pyridin-3-yl)methanol(15-6)

A mixture of [2-chloro-4-(1,4-dimethyl-1H-pyrazol-5-yl)-6-[(3R)-3-methylmorpholin-4-yl]pyridin-3-yl]methanol (135 mg, 0.40 mmol), 1-({[2-(trimethylsilyl)ethoxy]methyl}-λ^2-chloroalyl)-1H-pyrazol-5-amine (128 mg, 0.60 mmol), BrettPhos-Pd-G3 (36 mg, 0.04 mmol), and _Cs₂CO₃ (392 mg, 1.20 mmol) in _dioxane (6 mL) was stirred overnight at 100 °C under a nitrogen atmosphere. LC-MS showed that the reaction was complete. The mixture was diluted with EA (60 mL), washed with water and brine, _dried over anhydrous _Na₂SO₄ , filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 1:1, V/V) to obtain the desired product (46 mg, yield: 22%). LC/MS (ESI): m/z 514 [M+H] ⁺ .

Step 6. (R)-(2-((1H-pyrazol-5-yl)amino)-4-(1,4-dimethyl-1H-pyrazol-5-yl)-6-(3-methylmorpholino)pyridin-3-yl)methanol (15)

A mixture of [4-(1,4-dimethyl-1H-pyrazol-5-yl)-6-[(3R)-3-methylmorpholin-4-yl]-2-{[1-({[2-(trimethylsilyl)ethoxy]methyl}-λ^2-chloroalyl)-1H-pyrazol-5-yl]amino}pyridin-3-yl]methanol (46 mg, 0.09 mmol) in TBAF solution (1.0 M in THF, 5 mL, 5.0 mmol) was stirred overnight at 40 °C. LC-MS showed the reaction was complete. The reaction mixture was diluted with EA (40 mL), washed with water and brine, _dried over anhydrous _Na₂SO₄ , filtered, and concentrated. The residue was purified by preparative HPLC (C18, 10-95% acetonitrile in _H₂O containing 0.1% ammonia) to give the desired product (13.2 mg, 38%). LC/MS(ESI)m/z:384[M+H] ⁺ . ¹ H NMR(400MHz,DMSO)δ12.16(s,1H),8.51(s,1H),7.59(s,1H),7.32(s,1H),6.55(s,1 H),5.89(s,1H),5.38(s,1H),4.26(s,1H),4.22–4.14(m,2H),3.96–3.90(m,1H),3.8 5–3.78(m,1H),3.72(d,J＝11.3Hz,1H),3.64(d,J＝2.6Hz,1H),3.61(d,J＝2.9Hz,1H) ,3.58(d,J＝1.6Hz,3H),3.10–3.03(m,1H),1.85(s,3H),1.14(dd,J＝6.6,3.0Hz,3H).

Example 16

Step 1.2, methyl 6-dichloro-4-methylpyridine-3-carboxylate (17-2)

_CH₃I (0.3 mL, 4.85 mmol) _{and K₂CO₃} (503 mg, 3.64 mmol) were added to a solution of 2,6-dichloro-4-methylpyridin-3-carboxylic acid (500 mg, 2.43 mmol) in DMF (10 mL). The mixture was stirred overnight at room temperature. _LC -MS showed that the reaction was complete. The reaction mixture was diluted with EA (60 mL), washed with water and brine, dried over anhydrous _Na₂SO₄ , filtered, and concentrated to dryness. The residue was purified by silica gel column chromatography (PE:EA = 10:1, V/V) to give the desired product (525 mg, yield: 98%). LC/MS (ESI): m/z 220 [M+H] ^⁺ .

Step 2.4-(bromomethyl)-2,6-dichloropyridine-3-carboxylate (17-3)

NBS (0.97 g, 5.45 mmol) and AIBN (74 mg, 0.45 mmol) were added to a solution of methyl 2,6-dichloro- _{4-methylpyridine-3-carboxylate (1 g, 4.54 mmol) in CCl4 (40 mL). The mixture was stirred overnight at 80 °C. LC-MS showed that the reaction was complete. The reaction mixture was diluted with EA (60 mL), washed with a saturated aqueous solution of Na₂S₂O₃ and brine , dried} over anhydrous _Na₂SO₄ , filtered, and concentrated to dryness. The residue was purified by silica gel column chromatography (PE:EA = 10:1, V/V) to give the desired product (1.04 g, yield: 76%). LC/MS (ESI): m/z 300 [M+H] ^⁺ .

Step 3.2, Methyl 6-dichloro-4-(methanesulfonylmethyl)pyridine-3-carboxylate (17-4)

Sodium methanesulfonate (410 mg, 4.01 mmol) was added to a solution of methyl 4-(bromomethyl)-2,6-dichloropyridine-3-carboxylate (600 mg, _2.00 mmol) in DMF (20 mL). The mixture was stirred at room temperature for 2 hours. LC-MS showed that the reaction was complete. The reaction mixture was diluted with EA (60 mL), washed with water and brine, dried over anhydrous _Na₂SO₄ , filtered, and concentrated to dryness. The residue was purified by silica gel column chromatography (PE:EA = 5:1, V/V) to give the desired product (510 mg, yield: 85%). LC/MS (ESI): m/z 298 [M+H] ^⁺ .

Step 4. 2-Chloro-4-(methanesulfonylmethyl)-6-[(3R)-3-methylmorpholin-4-yl]pyridine-3-carboxylic acid methyl ester (17-6)

(3R)-3-methylmorpholine (204 mg, 2.01 mmol) was added to a solution of methyl 2,6-dichloro-4-(methanesulfonylmethyl)pyridine-3-carboxylate (300 mg, 1.01 mmol) in NMP (9 mL). The mixture was stirred for 1 hour at 120 °C under microwave irradiation. _LC -MS showed that the reaction was complete. The reaction mixture was diluted with EA (60 mL), washed with water and brine, dried over anhydrous _Na₂SO₄ , filtered, and concentrated to dryness. The residue was purified by silica gel column chromatography (PE:EA = 3:1, V/V) to give the desired product (150 mg, yield: 41%). LC/MS (ESI): m/z 363 [M+H] ^⁺ .

Step 5. 2-Chloro-4-(1-Methanesulfonylcyclopropyl)-6-[(3R)-3-methylmorpholin-4-yl]pyridine-3-carboxylic acid methyl ester (17-7)

To a solution of methyl 2-chloro-4-(methanesulfonylmethyl)-6-[(3R)-3-methylmorpholin-4-yl]pyridine-3-carboxylate (150 mg, 0.41 mmol) in toluene (20 mL), TBAB (27 mg, 0.08 mmol), 1,2-dibromoethane (233 mg, 1.24 mmol), and an aqueous solution of NaOH (10 M, 0.41 mL, _4.13 mmol) were added. The mixture was stirred at 60 °C for 6 hours. LC-MS showed the reaction was complete. The reaction mixture was diluted with EA (60 mL), washed with water and brine, dried over anhydrous _Na₂SO₄ , filtered, and concentrated to dryness. The residue was purified by silica gel column chromatography (PE:EA = 3:1, V/V) to give the desired product (100 mg, yield: 62%). LC/MS (ESI): m/z 389 [M+H] ^⁺ .

Step 6. [2-Chloro-4-(1-Methanesulfonylcyclopropyl)-6-[(3R)-3-methylmorpholin-4-yl]pyridin-3-yl]methanol (17-8)

LiBH₄ ( _2M in THF, 1.03 mL, 2.06 mmol) was added to a solution of methyl 2-chloro-4-(1-methanesulfonylcyclopropyl)-6-[(3R)-3-methylmorpholin-4-yl]pyridine-3-carboxylate (200 mg, 0.51 _mmol ) in 10 mL of THF. The mixture was stirred overnight at room temperature. LC-MS showed that the reaction was complete. The reaction mixture was diluted with EA (60 mL), washed with water and brine, dried over anhydrous _Na₂SO₄ , filtered, and concentrated to dryness. The residue was purified by silica gel column chromatography (PE:EA = 1:1, V/V) to give the desired product (150 mg, yield: 80%). LC/MS (ESI): m/z 361 [M+H] ^⁺ .

Step 7. (R)-(2-((1H-pyrazol-5-yl)amino)-6-(3-methylmorpholino)-4-(1-(methanesulfonyl)cyclopropyl)pyridin-3-yl)methanol (17)

Brettphos-Pd-G3 (12.5 mmol, 0.014 mmol) and Cs₂CO₃ (135 mg, 0.41 mmol) were added to a solution of (R)-(2-chloro-6-(3-methylmorpholino)-4-(1-(methanesulfonyl)cyclopropyl)pyridin-3-yl)methanol (50 mg, 0.14 mmol) and _1H -pyrazol-5-amine (23 mg, 0.28 mmol) in dioxane ( ₂ mL). The mixture was stirred overnight at 110 °C under a _nitrogen atmosphere. _LC -MS showed that the reaction was complete. The reaction mixture was diluted with EA (40 mL), washed with water and brine, dried over anhydrous _Na₂SO₄ , filtered, and concentrated to dryness. The residue was purified by preparative HPLC ( _C18 , 10-95% MeOH in _H2O , containing 0.1% HCOOH) to obtain the desired product (14 mg, yield: 24%). LC/MS (ESI): m/z 408 [M+H] ⁺ . ¹ H NMR (400MHz, DMSO) δ 12.11 (s, 1H), 8.52 (d, J = 4.3Hz, 1H), 7.56 (d, J = 1.6Hz, 1H), 6.50 (s, 1H), 6.25 (s, 1H), 5.30 (s, 1H), 4.89 (d, J = 13.3Hz, 1H), 4.35 (d, J = 12.8Hz, 1H), 4.29 (s, 1H), 3 .94(d,J＝8.4Hz,1H),3.77(dd,J＝25.7,11.3Hz,2H),3.65(s,2H),3.08(d,J＝11.5Hz,1H),2 .96(s,3H),1.84(s,1H),1.49(d,J＝28.1Hz,2H),1.33(s,1H),1.13(dd,J＝18.8,5.4Hz,3H).

Example 17

Step 1.5-{[4-(1-methanesulfonylcyclopropyl)-6-[(3R)-3-methylmorpholin-4-yl]pyridin-2-yl]amino}-3-methyl-1H-pyrazole-1-carboxylic acid tert-butyl ester (19-2)

A mixture of (3R)-4-[6-chloro-4-(1-methanesulfonylcyclopropyl)pyridin-2-yl]-3-methylmorpholine (450 mg, 1.36 mmol), tert-butyl 5-amino-3-methyl-1H-pyrazole-1-carboxylate (402 mg, 2.04 mmol), BrettPhos-Pd-G3 (27 mg, 0.03 mmol), _{and Cs₂CO₃} (1.1 g, 3.40 mmol) in dioxane (40 mL) was stirred overnight at 100 ° _C under a nitrogen atmosphere. LC-MS showed the reaction was complete. The mixture was diluted with EA (50 mL), washed with water and brine, dried over anhydrous _Na₂SO₄ , filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 1:1, V/V) to give the desired product (526 mg, yield: 79%). LC/MS ESI(m/z):492[M+H] ⁺ .

Step 2.4-(1-Methanesulfonylcyclopropyl)-N-(3-methyl-1H-pyrazol-5-yl)-6-[(3R)-3-methylmorpholin-4-yl]pyridine-2-amine (19)

A mixture of 5-{[4-(1-methanesulfonylcyclopropyl)-6-[(3R)-3-methylmorpholin-4-yl]pyridin-2-yl]amino}-3-methyl-1H-pyrazole-1-carboxylic acid tert-butyl ester (526 mg, 1.07 mmol) in HCl solution (4 M in dioxane, 8 mL) was stirred overnight at room temperature. LC-MS showed the reaction was complete. The mixture was concentrated to dryness under reduced pressure. The residue was purified by preparative HPLC ( _C18 , 10-95% MeOH in _H2O containing 0.1% HCOOH) to give the desired product (213 mg, yield: 51%). LC/MS ESI (m/z): 392 [M+H] ⁺ . ¹ H NMR (400MHz, DMSO) δ11.69(s,1H),8.79(s,1H),6.66(s,1H),6.18(s,1H),6.00(s,1H),4.26(d, J＝6.6Hz,1H),3.93(dd,J＝11.2,3.2Hz,1H),3.81(d,J＝11.1Hz,1H),3.72(d,J＝11.2Hz,1H),3.6 1(dd,J＝11.3,2.8Hz,1H),3.47(dd,J＝11.6,8.9Hz,1H),3.04(td,J＝12.5,3.6Hz,1H),2.93(s,3 H), 2.17 (s, 3H), 1.56 (dd, J = 5.7, 3.9Hz, 2H), 1.24 (dd, J = 6.2, 4.7Hz, 2H), 1.13 (d, J = 6.6Hz, 3H).

Example 18

Step 1.2, 6-Dichloro-4-iodo-3-methylpyridine (20-2)

At -60 °C, LDA (2 M in THF, 5.48 mL, 10.95 mmol) was added dropwise to a solution of 2,6-dichloro-3-iodopyridine (2 g, 7.30 mmol) in DMF (40 mL). The mixture was stirred at -60 °C for 1 hour, followed by the dropwise addition of iodomethane (0.68 mL, 10.95 mmol). The resulting mixture was stirred at _{-60 °C} for another hour. LC-MS showed the reaction was complete. The reaction was quenched with a saturated aqueous solution of _NH4Cl and extracted with EA (50 mL). The organic layer was separated, washed with brine, dried over anhydrous _Na2SO4 , filtered, and concentrated. The residue was purified by silica gel chromatography (PE:EA = 10:1, V/V) to give the desired product (1.7 g, yield: 81%). LC/MS (ESI): m/z 288 [M+H] ⁺ .

Step 2. (R)-4-(6-chloro-4-iodo-5-methylpyridin-2-yl)-3-methylmorpholine (20-4)

(R)-3-methylmorpholine (1.79 g, 17.71 mmol) and N,N-diisopropylethylamine (2.93 mL, 17.71 mmol) were added to a solution of 2,6-dichloro-4-iodo-3-methylpyridine (1.7 g, 5.90 mmol) in _NMP (17.0 mL). The mixture was stirred for 1 hour at 180 °C under microwave irradiation. LC-MS showed that the reaction was complete. The mixture was diluted with EA (60 mL), washed with water and brine, dried over anhydrous _Na₂SO₄ , filtered, and concentrated. The residue was purified by silica gel rapid chromatography (PE:EA = 3:1, V/V) to give the desired product (750 mg, yield: 36%). LC/MS (ESI): m/z 353 [M+H] ^⁺ .

Step 3. (R)-4-(6-chloro-4-(1,4-dimethyl-1H-1,2,3-triazol-5-yl)-5-methylpyridin-2-yl)-3-methylmorpholine (20-6)

To a solution of (R)-4-(6-chloro-4-iodo-5-methylpyridin-2-yl)-3-methylmorpholine (600 mg, 1.70 mmol) in DMF (6 mL), 1,4-dimethyl-1H-1,2,3-triazole (182 mg, 1.87 mmol), tetramethylammonium acetate (272 mg, 2.04 mmol), and bis(triphenylphosphine)palladium(II) chloride (132 mg, 0.17 mmol) were added. The mixture was stirred for 5 hours at 140 °C under a nitrogen atmosphere. _LC -MS showed that the reaction was complete. The mixture was diluted with EA (60 mL), washed with water and brine, dried over anhydrous _Na₂SO₄ , filtered, and concentrated. The residue was purified by silica gel rapid chromatography (PE:EA = 1:1, V/V) to give the desired product (400 mg, yield: 73%). LC/MS(ESI): m/z 322[M+H] ⁺ .

Step 4. (R)-4-(1,4-dimethyl-1H-1,2,3-triazol-5-yl)-3-methyl-6-(3-methylmorpholino)-N-(1H-pyrazol-5-yl)pyridin-2-amine (20)

To a solution of (R)-4-(6-chloro-4-(1,4-dimethyl-1H-1,2,3-triazol-5-yl)-5-methylpyridin-2-yl)-3-methylmorpholine (100 mg, 0.31 mmol) in dioxane (2 mL), tert-butyl 5-amino-1H-pyrazole-1 _- carboxylate (85 mg, 0.47 mmol), _Cs₂CO₃ (203 mg, 0.62 mmol), and BrettPhos-Pd-G₃ (28 mg, 0.03 mmol) were added. The mixture was stirred overnight at 100 °C under a nitrogen atmosphere. LC-MS showed the reaction was complete. The mixture was diluted with EA (60 mL), washed with water and brine, _dried over anhydrous _Na₂SO₄ , filtered, and concentrated. The residue was dissolved in DCM (5 mL), and then HCl solution (4 M in dioxane, 2 mL) was added. The resulting mixture was stirred at room temperature for 12 hours. LC-MS showed the reaction was complete. The mixture was concentrated to dryness under reduced pressure. The residue was purified by preparative HPLC (C18, 10-95% MeOH in _H₂O containing 0.1% HCOOH) to give the desired product (10 mg, yield: 8.7%). LC/MS (ESI): m/z 369 [M+H] ⁺ . ¹ HNMR(400MHz,DMSO)δ8.13(s,1H),7.56(s,1H),6.46(s,1H),5.97(s,1H),4. 24–4.14(m,1H),3.90(dd,J＝11.2,3.1Hz,1H),3.79(s,3H),3.77–3.71(m,1H) ,3.69(d,J＝11.5Hz,1H),3.61(d,J＝10.8Hz,1H),3.46(d,J＝2.6Hz,1H),3.02 (t,J＝12.5Hz,1H),2.10(s,3H),1.82(d,J＝0.8Hz,3H),1.11(d,J＝6.6Hz,3H).

Example 19

Step 1. (R)-4-(1,4-dimethyl-1H-1,2,3-triazol-5-yl)-3-methyl-N-(3-methyl-1H-pyrazol-5-yl)-6-(3-methylmorpholino)pyridine-2-amine (21)

Add 3-methyl-1H-pyrazole-5-amine (45 mg, 0.47 mmol), Cs₂CO₃ (203 mg, 0.62 mmol), and BrettPhos-Pd- _G₃ (28 mg, 0.03 mmol) to a solution of (R)-4-(6-chloro-4-(1,4-dimethyl-1H-1,2,3-triazol-5-yl)-5-methylpyridin- ₂ -yl)-3-methylmorpholine (100 mg, 0.31 mmol) in dioxane (2 mL). Stir the mixture overnight at 100 °C under a nitrogen atmosphere. LC-MS showed the reaction was complete. Dilute the mixture with EA (60 mL), wash with water and brine, _dry over anhydrous _Na₂SO₄ , filter, and concentrate. Dissolve the residue in DCM (5 mL), then add HCl solution (4 M in dioxane, 2 mL). Stir the resulting mixture at room temperature for 12 hours. LC-MS showed that the reaction was complete. The mixture was concentrated to dryness under reduced pressure. The residue was purified by preparative HPLC (C18, 10-95% MeOH in _H₂O , containing 0.1% HCOOH) to give the desired product (15 mg, yield: 12.6%). LC/MS(ESI)m/z:383[M+H] ⁺ . ¹ HNMR (400MHz, DMSO) δ8.00(s,1H),6.22(s,1H),5.96(s,1H),4.20(s,1H),3.91(dd,J＝11.2,3.1Hz,1H),3.78(s,3H),3.77–3.72(m,1H),3.70(d,J＝ 11.8Hz,1H),3.61(d,J＝10.8Hz,1H),3.50–3.43(m,1H),3.06–2.98(m,1H ), 2.20 (s, 3H), 2.09 (s, 3H), 1.80 (d, J = 0.7Hz, 3H), 1.12 (d, J = 6.6Hz, 3H).

Example 20

Step 1: (R)-4-(4,6-dichloropyridin-2-yl)-3-methylmorpholine (22-2)

DIEA (755 mg, 5.84 mmol) was added to a solution of 2,6-dichloro-4-iodopyridine (800 mg, 2.92 mmol) and (R)-3-methylmorpholine (325 mg, 3.21 mmol) in DMA (8 mL). _N₂ was injected twice into the mixture, and the mixture was stirred at 120 °C for 12 h. The reaction mixture was diluted with water (30 mL) and extracted with EA (30 mL × 2). The combined organic layers were washed with brine (50 mL), dried over _{anhydrous Na₂SO₄} , filtered, and concentrated under vacuum. The resulting mixture was purified by rapid chromatography using PE/EtOAc (20:1, 8:1) elution to give the desired product (500 mg, yield: 68.9%).

Step 2: (R)-4-(6-chloro-4-(3,5-dimethylisoxazol-4-yl)pyridin-2-yl)-3-methylmorpholine (22-3)

To a solution of (R)-4-(6-chloro-4-iodopyridin-2-yl)-3-methylmorpholine (300 mg, 1.21 mmol) and 3,5-dimethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxane-2-yl)isoxazole (270.8 mg, 1.21 mmol) in dioxane (10 mL) _{, Na₂CO₃} (320 mg, 3.03 mmol) and Pd(dppf) _Cl₂ (88 mg, 0.12 mmol) were added. The mixture was injected twice with _N₂ and stirred at 90 °C for 12 hours. The reaction mixture was diluted with water (30 mL) and extracted with EA (30 mL × 2). The combined organic layers were washed with brine (50 mL), dried over _{anhydrous Na₂SO₄} , filtered, and concentrated under vacuum. The resulting mixture was purified by rapid chromatography using PE/EtOAc (3:1, 1:1) elution to obtain the desired product (320 mg, yield: 86.02%).

Step 3: (R)-4-(3,5-dimethylisoxazol-4-yl)-6-(3-methylmorpholino)-N-(1H-pyrazol-5-yl)pyridin-2-amine (22)

Cs₂CO₃ (400 mg, 1.23 mmol) and BrettPhos Pd G₃ (45 mg, 0.049 mmol) were added to dioxane (8 mL) containing (R)-4-(6-chloro-4-(3,5-dimethylisoxazol-4-yl)pyridin-2-yl) _{-3- methylmorpholine} (150 mg, 0.49 mmol) and tert-butyl 5-amino-1H-pyrazol-1-carboxylate (108 mg, 0.59 mmol). The mixture was injected twice with _N₂ and stirred overnight at 90 °C. The reaction mixture was diluted with water and extracted with EtOAc (30 mL × 2). The combined organic layers were washed with brine (50 mL), dried over _{anhydrous Na₂SO₄} , filtered, and concentrated under vacuum. The residue was purified by preparative TLC (DCM/MeOH = 10/1) to obtain the desired product (20 mg, yield: 11.49%). LC/MS (ESI) m/z: 355.0 [M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d6) δppm 12.03 (br s, 1H) 8.98 (br s, 1H) 7.55 (br s, 1H) 6.33-6.47 (m, 2H) 5.99 (s, 1H) 4.30 (br d, J＝5.16Hz, 1H) 3.94 (br dd,J＝11.12,3.20Hz,1H)3.84(br d,J＝11.68Hz,1H)3.71-3.75(m,1H)3.61-3.66(m,1H)3.45-3.52(m,1H)3.03-3.10(m,1H)2.44(s,3H)2.25(s,3H)1.15(d,J＝6.64Hz,3H).

Example 21

Step 1. (3R)-4-(6-chloro-4-iodopyridin-2-yl)-3-methylmorpholine (23-3)

(3R)-3-methylmorpholine (554.1 mg, 5.48 mmol) was added to a solution of 2,6-dichloro-4-iodopyridine (500 mg, 1.83 mmol) in NMP ( _{10 mL). The mixture was stirred for 1 hour at 150 °C under microwave irradiation. The reaction mixture was diluted with EA (60 mL), washed with water and brine, dried over anhydrous Na₂SO₄ ,} filtered, and concentrated to dryness. The residue was purified by silica gel column chromatography (PE:EA = 10:1, V/V) to give the desired product (250 mg, yield: 40%). LC/MS (ESI): m/z 339 [M+H] ^⁺ .

Step 2. (3R)-4-[6-chloro-4-(dimethyl-1H-1,2,3-triazol-5-yl)pyridin-2-yl]-3-methylmorpholine (23-5)

Pd(PPh3)2Cl2 (62.2 mg, 0.09 mmol) and tetramethylammonium acetate (141.6 mg, 1.06 mmol) were added to a solution of (3R)-4-(6-chloro-4-iodopyridin-2-yl) _{-3 -} methylmorpholine (300 mg, 0.88 mmol) and 1,4-dimethyl-1H-1,2,3-triazole (103.3 mg, 1.06 mmol) in DMF ( _{15 mL )} . The mixture was stirred overnight at 100 °C under a nitrogen atmosphere. LC-MS showed that the reaction was complete. The reaction mixture was diluted with DCM (60 mL), washed with water and brine, dried over anhydrous _Na2SO4 , filtered, and concentrated to dryness. The residue was purified by silica gel column chromatography (PE:EA = 3:1, V/V) to give the desired product (210 mg, yield: 77%). LC/MS(ESI): m/z 309[M+H] ⁺ .

Step 3.4-(dimethyl-1H-1,2,3-triazol-5-yl)-N-(3-methyl-1H-pyrazol-5-yl)-6-[(3R)-3-methylmorpholin-4-yl]pyridin-2-amine (23)

To a solution of (3R)-4-[6-chloro-4-(dimethyl-1H-1,2,3-triazol-5-yl)pyridin-2-yl]-3-methylmorpholine (90 mg, 0.29 mmol) in dioxane (2 mL), tert-butyl 5-amino-3-methyl-1H-pyrazole-1-carboxylate ( _86.51 mg, 0.439 mmol), BrettPhos-Pd-G3 (26.5 mg, 0.03 mmol), and _Cs₂CO₃ (190.5 mg, 0.59 mmol) were added. The mixture was stirred overnight at 100 °C under a nitrogen atmosphere. LC-MS showed that the reaction was complete. The reaction mixture was diluted with EA (40 mL), washed with water and brine, _dried over anhydrous _Na₂SO₄ , filtered, and concentrated to dryness. The residue was purified by preparative HPLC (C18, 10-95% MeCN in _H₂O , containing 0.1% ammonia) to obtain the desired product (35 mg, yield: 32%). LC/MS (ESI): m/z 369 [M+H] ⁺ . ¹ H NMR (400MHz, DMSO) δ11.71(s,1H),8.92(s,1H),6.55(s,1H),6.07(d,J＝6.1Hz ,2H),4.31(d,J＝5.9Hz,1H),3.96(s,3H),3.90(dd,J＝14.1,8.1Hz,2H),3.72( d,J＝11.2Hz,1H),3.63(dd,J＝11.2,2.9Hz,1H),3.48(td,J＝11.8,2.8Hz,1H), 3.07(td,J=12.5,3.5Hz,1H),2.25(s,3H),2.18(s,3H),1.16(d,J=6.6Hz,3H).

Example 22

Step 1. (R)-2-(2-((1H-pyrazol-5-yl)amino)-6-(3-methylmorpholino)pyridin-4-yl)-2-methylpropionitrile (24)

BrettPhos-Pd-G3 (19 mg, 0.21 mmol) and Cs₂CO₃ (210 mg, 0.64 mmol) were added to a solution containing 2-{2-chloro-6-[(3R)-3-methylmorpholin-4-yl]pyridin-4-yl}-2-methylpropionitrile (60 mg, 0.21 mmol) and 1H-pyrazol- _{5 -} amine (35 mg, 0.42 mmol) in dioxane (3 _mL ). The mixture was stirred for 10 hours at 110 °C under a _nitrogen atmosphere. LC-MS showed that the reaction was complete. The reaction mixture was diluted with EA (40 mL), washed with water and brine, dried over anhydrous _Na₂SO₄ , filtered, and concentrated. The residue was purified by preparative HPLC ( _C₁₈ , 10-95% MeOH in _H₂O containing 0.1% HCOOH) to give the desired product (60 mg, yield: 85%). LC/MS(ESI):m/z 327[M+H] ⁺ . ¹ H NMR (400MHz, DMSO) δ12.10 (s, 1H), 9.02 (s, 1H), 7.53 (d, J = 2.2Hz, 1H), 6.59 (s, 1H), 6.30 (d,J＝1.9Hz,1H),6.08(d,J＝1.0Hz,1H),4.32(d,J＝6.5Hz,1H),3.93(dd,J＝11.2,3.3Hz, 1H),3.79(d,J＝12.8Hz,1H),3.73(d,J＝11.2Hz,1H),3.62(dd,J＝11.3,2.9Hz,1H),3.47( td,J＝11.8,3.0Hz,1H),3.06(td,J＝12.6,3.7Hz,1H),1.63(s,6H),1.13(d,J＝6.6Hz,3H).

Example 23

Step 1.2-{2-chloro-6-[(3R)-3-methylmorpholin-4-yl]pyridin-4-yl}-2-methylpropionitrile (25-2)

At 0 °C, sodium tert-butoxide (198 mg, 2.06 mmol) and methyl iodide (0.13 mL, 2.06 mmol) were added to a solution of 2-{2-chloro-6-[(3R)-3-methylmorpholin-4-yl]pyridin-4-yl}acetonitrile (173 mg, 0.69 mmol) in THF (6 mL). The mixture was stirred overnight at ambient temperature. _LC -MS showed that the reaction was complete. The mixture was diluted with EA (50 mL), washed with water and brine, dried over anhydrous _Na₂SO₄ , filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 3:1, V/V) to give the desired product (154 mg, yield: 80%). LC/MS ESI (m/z): 280 [M+H] ^⁺ .

Step 2.5-{[4-(1-cyano-1-methylethyl)-6-[(3R)-3-methylmorpholin-4-yl]pyridin-2-yl]amino}-3-methyl-1H-pyrazole-1-carboxylic acid tert-butyl ester (25-3)

BrettPhos-Pd-G3 (50 mg, 0.06 mmol) and Cs₂CO₃ (538 mg, 1.65 mmol) were added to a solution of 2-{2-chloro-6-[(3R)-3-methylmorpholin-4-yl]pyridin-4-yl}-2-methylpropionitrile (154 mg, 0.55 mmol) and tert-butyl _5- amino- ₃ -methyl-1H-pyrazol-1-carboxylate (163 mg, 0.83 mmol) in dioxane (10 mL). The mixture was stirred overnight at 100 °C under a nitrogen atmosphere. _LC -MS showed that the reaction was complete. The mixture was diluted with EA (50 mL), washed with water and brine, dried over anhydrous _Na₂SO₄ , filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 1:1, V/V) to give the desired product (156 mg, yield: 64%). LC/MS ESI(m/z):441[M+H] ⁺ .

Step 3.2-Methyl-2-{2-[(3-methyl-1H-pyrazol-5-yl)amino]-6-[(3R)-3-methylmorpholin-4-yl]pyridin-4-yl}propionitrile (25)

A mixture of 5-{[4-(1-cyano-1-methylethyl)-6-[(3R)-3-methylmorpholin-4-yl]pyridin-2-yl]amino}-3-methyl-1H-pyrazole-1-carboxylic acid tert-butyl ester (156 mg, 0.35 mmol) in HCl solution (4 M in dioxane, 4 mL) was stirred overnight at room temperature. LC-MS showed the reaction was complete. The mixture was concentrated to dryness under reduced pressure. The residue was purified by preparative HPLC ( _C18 , 10-95% MeOH in _H2O containing 0.1% HCOOH) to give the desired product (88.5 mg, yield: 73%). LC/MS ESI (m/z): 341 [M+H] ⁺ . ¹ H NMR(400MHz,DMSO)δ9.43(s,1H),6.60(s,1H),6.18(s,1H),6.04(s,1H),4.30(d d,J＝6.6,1.9Hz,1H),3.95(dd,J＝11.3,3.4Hz,1H),3.80(dd,J＝13.1,2.0Hz,1H), 3.74(d,J＝11.3Hz,1H),3.64(dd,J＝11.3,2.8Hz,1H),3.49(td,J＝11.8,3.0Hz,1H ), 3.11(td,J＝12.6,3.8Hz,1H),2.21(s,3H),1.64(s,6H),1.15(d,J＝6.6Hz,3H).

Example 24

Step 1. (R)-4-(6-chloro-4-(2-(methanesulfonyl)prop-2-yl)pyridin-2-yl)-3-methylmorpholine (26-1)

_CH₃I (4.7 mL, 76.11 mmol) and t-BuONa (7.31 g, 76.11 mmol) were added to a solution of (3R)-4-[6-chloro-4-(methanesulfonylmethyl)pyridin-2-yl]-3-methylmorpholine (5.8 g, 19.03 mmol) in THF (100 mL). The reaction mixture was stirred overnight at room temperature. The reaction mixture was diluted with EA (100 mL), washed with water and brine, _dried over anhydrous _Na₂SO₄ , filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 1:1, V/V) to give the desired product (5.3 g, yield: 83.7%). LC/MS (ESI): m/z 333 [M+H] ^⁺ .

Step 2.6-((R)-3-methylmorpholino)-4-(2-(methanesulfonyl)propyl-2-yl)-N-(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-5-yl)pyridin-2-amine (26-3)

To a solution of (3R)-4-[6-chloro-4-(2-methanesulfonylprop-2-yl)pyridin- _2- yl]-3-methylmorpholine (4.0 g, 12.02 mmol) in dioxane (80 mL), 1-(tetrahydropyran-2-yl)-1H-pyrazol-5-amine (3.0 g, 18.03 mmol), Brettphos Pd G3 (1.09 g, 1.20 _mmol ), and _Cs₂CO₃ (11.8 g, 36.05 mmol) were added. The mixture was stirred overnight at 100 °C under a nitrogen atmosphere. The reaction mixture was diluted with DCM (100 mL), washed with water and brine, dried over anhydrous _Na₂SO₄ , filtered, and concentrated. The residue was purified by silica gel column chromatography (DCM:MeOH = 30:1, V/V) to give the desired product (4.45 g, yield: 80%). LC/MS(ESI): m/z 464[M+H] ⁺ .

Step 3. (R)-6-(3-methylmorpholino)-4-(2-(methanesulfonyl)propyl-2-yl)-N-(1H-pyrazol-5-yl)pyridin-2-amine (26)

HCl/dioxane (50 mL) was added to a solution of 4-(2-methanesulfonylprop-2-yl)-6-[(3R)-3-methylmorpholin-4-yl]-N-[1-(tetrahydropyran-2-yl)-1H-pyrazol-5-yl]pyridine- ₂ -amine (4.45 g, 9.60 mmol) in DCM (50 mL). The reaction was stirred overnight at room temperature. The reaction mixture was concentrated under vacuum. The residue was diluted with DCM (50 mL), washed with saturated aqueous NaHCO3 solution and brine, dried over _{anhydrous Na2SO4} , filtered, and concentrated. The residue was purified by preparative HPLC ( _C18 , 10-95% MeOH in _H2O containing 0.1% HCOOH) to give the desired product (2.08 g, 57%). LC/MS (ESI): m/z 380 [M+H] ⁺ . ¹ H NMR(400MHz,DMSO)δ12.02(s,1H),8.95(s,1H),7.53(s,1H),6.65(s,1H),6.27(d, J＝60.7Hz,1H),6.19(s,1H),4.28(d,J＝6.2Hz,1H),3.94(dd,J＝11.1,3.2Hz,1H),3. 76(dd,J＝22.8,11.2Hz,2H),3.63(dd,J＝11.2,2.8Hz,1H),3.48(td,J＝11.7,2.9Hz, 1H), 3.05 (td, J = 12.6, 3.7Hz, 1H), 2.76 (s, 3H), 1.66 (s, 6H), 1.12 (d, J = 6.6Hz, 3H).

Example 25

Step 1.5-{[4-(2-methanesulfonylprop-2-yl)-6-[(3R)-3-methylmorpholin-4-yl]pyridin-2-yl]amino}-3-methyl-1H-pyrazole-1-carboxylic acid tert-butyl ester (27-1)

A mixture of (3R)-4-[6-chloro-4-(2-methanesulfonylprop-2-yl)pyridin-2-yl]-3-methylmorpholine (580 mg, 1.74 mmol), tert-butyl 5-amino-3-methyl-1H-pyrazole-1-carboxylate (516 mg, 2.61 mmol), BrettPhos-Pd-G3 (157.9 mg, 0.17 mmol ₎ , and _Cs₂CO₃ (1.42 g, 4.36 mmol) in dioxane (20 mL) was stirred overnight at 100 °C under a nitrogen atmosphere. _LC -MS showed the reaction was complete. The mixture was diluted with EA (50 mL), washed with water and brine, dried over anhydrous _Na₂SO₄ , filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 1:1, V/V) to give the desired product (758 mg, yield: 88%). LC/MS ESI(m/z):494[M+H] ⁺ .

Step 2.4-(2-methanesulfonylprop-2-yl)-N-(3-methyl-1H-pyrazol-5-yl)-6-[(3R)-3-methylmorpholin-4-yl]pyridine-2-amine (27)

At ambient temperature, a mixture of 5-{[4-(2-methanesulfonylprop-2-yl)-6-[(3R)-3-methylmorpholin-4-yl]pyridin-2-yl]amino}-3-methyl-1H-pyrazole-1-carboxylic acid tert-butyl ester (758 mg, 1.54 mmol) in HCl solution (4 M in dioxane, 8 mL) was stirred overnight. LC-MS showed the reaction was complete. The mixture was concentrated to dryness under reduced pressure. The residue was purified by preparative HPLC ( _C18 , 10-95% MeOH in _H2O containing 0.1% HCOOH) to give the desired product (215 mg, yield: 35%). LC/MS ESI (m/z): 394 [M+H] ⁺ . ¹ H NMR(400MHz,DMSO)δ11.67(s,1H),8.79(s,1H),6.70(s,1H),6.19(s,1H),6.03(s,1H ),4.27(d,J＝6.6Hz,1H),3.94(dd,J＝11.2,3.2Hz,1H),3.79(d,J＝13.1Hz,1H),3.73( d,J＝11.2Hz,1H),3.63(dd,J＝11.2,2.8Hz,1H),3.48(td,J＝11.8,2.9Hz,1H),3.04(t d,J＝12.6,3.7Hz,1H),2.75(s,3H),2.17(s,3H),1.66(s,6H),1.12(d,J＝6.6Hz,3H).

Example 26

Step 1. (R)-2-(2-((1H-pyrazol-5-yl)amino)-6-(3-methylmorpholino)pyrimidin-4-yl)-2-methylpropionitrile (28)

A mixture of 2-{2-chloro-6-[( _3R )-3-methylmorpholin-4-yl]pyrimidin-4-yl}-2-methylpropionitrile (100 mg, 0.35 mmol), 1H-pyrazol-5-amine (59 mg, 0.71 mmol), BrettPhos Pd G3 (32 mg, 0.03 mmol), and _Cs₂CO₃ (349 mg, 1.07 mmol) in _dioxane (4 mL) was stirred for 16 hours at 110 °C under a _N₂ atmosphere. LC-MS showed the reaction was complete. The reaction mixture was diluted with EA (40 mL), washed with water and brine, dried over anhydrous _Na₂SO₄ , filtered, and concentrated under vacuum. The residue was purified by preparative HPLC ( _C₁₈ , 10-95% MeOH in _H₂O containing 0.1% HCOOH) to give the desired product (36 mg, yield: 31%). LC/MS(ESI):m/z 328[M+H] ⁺ . ¹ H NMR(400MHz,DMSO)δ12.08(s,1H),9.20(s,1H),8.13(s,1H),7.51(s,1H),6.52 (s,1H),6.25(s,1H),4.41(s,1H),4.01(d,J＝12.8Hz,1H),3.93(dd,J＝11.3,3. 4Hz, 1H), 3.72 (d, J = 11.4Hz, 1H), 3.58 (dd, J = 11.4, 3.0Hz, 1H), 3.43 (td, J = 11. 8, 2.9Hz, 1H), 3.14 (td, J = 13.0, 3.8Hz, 1H), 1.65 (s, 6H), 1.19 (d, J = 6.7Hz, 3H).

Example 27

Step 1.2, 6-Dichloro-3-fluoro-4-iodopyridine (33-2)

At -78°C under a _nitrogen atmosphere, LDA (2.0 M in THF, 6.6 mL, 13.2 mmol) was added dropwise to a solution of 2,6-dichloro-3-fluoropyridine (2.0 g, 12.05 mmol) in anhydrous THF (30 mL). The mixture was stirred at -78°C for 1 hour, followed by the dropwise addition of _I₂ (4.0 g, 15.74 mmol) in anhydrous THF (10 mL). The resulting mixture was stirred at -78°C for another 1 hour. LC-MS showed the reaction was complete. The reaction mixture was quenched with a saturated aqueous _NH₄Cl solution and diluted with EA (30 mL _× 3). The combined organic layers were washed with a saturated _{aqueous Na₂S₂O₃} solution and brine, dried over anhydrous _{Na₂SO₄ ,} filtered, and concentrated under vacuum. The residue was purified by silica gel column chromatography (PE) to give the desired product (2.79 g, yield: 79%). ¹ H NMR (400MHz, DMSO) δ 8.16 (d, J=3.5Hz, 1H).

Step 2.2, 6-Dichloro-4-(1,4-dimethyl-1H-pyrazol-5-yl)-3-fluoropyridine (33-4)

A mixture of 2,6-dichloro- ₃ -fluoro-4-iodopyridine (1.0 g, 3.42 mmol), (1,4-dimethyl-1H-pyrazol-5-yl)boronic acid (0.76 g, 3.43 mmol), _PdCl₂ (dppf) (251 mg, 0.34 mmol), and _Na₂CO₃ (2.0 _M in _H₂O , 3.4 mL) in DME (35 mL) was stirred for 15 hours at 90 °C under a N₂ atmosphere. _LC -MS showed the reaction was complete. The reaction mixture was diluted with EA (40 mL), washed with water and brine, dried over anhydrous _Na₂SO₄ , filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 10:1, V/V) to give the desired product (744 mg, yield: 83%). LC/MS (ESI): m/z 260 [M+H] ^⁺ .

Step 3.6-Chloro-4-(1,4-dimethyl-1H-pyrazol-5-yl)-3-fluoro-N-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-5-yl)pyridine-2-amine (33-6)

A mixture of 2,6-dichloro- _4- (1,4-dimethyl-1H-pyrazol-5-yl)-3-fluoropyridine (400 mg, 1.53 mmol), 1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-pyrazol-5-amine (329 mg, 1.54 mmol), _Pd₂ (dba) _₃ (141 mg, 0.15 mmol), XantPhos (89 mg, 0.15 mmol), and _Cs₂CO₃ (1.0 g, 3.06 mmol) in _dioxane (25 mL) was stirred for 6 hours at 100 °C under a N₂ atmosphere. _LC -MS showed that the reaction was complete. The reaction mixture was diluted with EA (40 mL), washed with water and brine, dried over anhydrous _Na₂SO₄ , filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 2:1, V/V) to obtain the desired product (419 mg, yield: 62%). LC/MS (ESI): m/z 437 [M+H] ⁺ .

Step 4. (R)-4-(1,4-dimethyl-1H-pyrazol-5-yl)-3-fluoro-6-(3-methylmorpholino)-N-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-5-yl)pyridine-2-amine (33-8)

A mixture of 6-chloro-4-(1,4-dimethyl-1H-pyrazol- ₅ -yl)-3-fluoro-N-(1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-pyrazol-5-yl)pyridin-2-amine (400 mg, 0.91 mmol), (3R)-3-methylmorpholine (278 mg, 2.74 mmol), _Pd₂ (dba) _₃ (168 mg, 0.18 mmol), RuPhos (171 mg, 0.36 mmol), and _Cs₂CO₃ ( _1.19 g, 3.65 mmol) in dioxane (40 mL) was stirred for 6 hours at 100 °C under a N₂ atmosphere. _LC -MS showed that the reaction was complete. The reaction mixture was diluted with EA (40 mL), washed with water and brine, dried over anhydrous _Na₂SO₄ , filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 2:1, V/V) to obtain the desired product (437 mg, yield: 95%). LC/MS (ESI): m/z 502 [M+H] ⁺ .

Step 5. (R)-4-(1,4-dimethyl-1H-pyrazol-5-yl)-3-fluoro-6-(3-methylmorpholino)-N-(1H-pyrazol-5-yl)pyridin-2-amine (33)

At 70 °C, a mixture of 4-(1,4-dimethyl-1H-pyrazol-5-yl)-3-fluoro-6-[(3R)-3-methylmorpholin-4-yl]-N-(1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-pyrazol-5-yl)pyridin-2-amine (417 mg, 0.83 mmol) in TBAF solution (1.0 M in THF, 8 mL, 8 mmol) was stirred for 5 hours. LC-MS showed that the reaction was complete. The reaction mixture was diluted with EA (40 mL), washed with water (20 mL × 2) and brine, dried over anhydrous _{Na₂SO₄ ,} filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 1:1, V/V) to obtain a brown solid (252 mg). The brown solid was further purified by preparative HPLC (C ₁₈ , 10-95% MeOH in H ₂ O, containing 0.1% HCOOH) to obtain the desired product (66.9 mg, yield: 21%). LC/MS(ESI):m/z 372[M+H] ⁺ . ¹ H NMR (400MHz, DMSO) δ12.18(s,1H),8.89(s,1H),7.59(s,1H),7.37(s,1H),6.51(s,1H),5.94(s,1H),4.20(d,J＝4.8Hz,1H),3.97–3.89( m,1H),3.77–3.69(m,5H),3.63(dd,J＝11.2,2.7Hz,1H),3.51(s,1H),3.05(td,J＝12.6,3.7Hz,1H),1.96(s,3H),1.13(d,J＝6.6Hz,3H).

Example 28

Step 1.2, 6-Dichloro-3-fluoroisononal (34-3)

At -78 °C, LDA (2.5 M in THF, 9.4 mL, 23.50 mmol) was added dropwise to a solution of 2,6-dichloro-3-fluoropyridine (3 g, 18.07 mmol) in THF (50 mL). The mixture was stirred at -78 °C for 1 hour, followed by the dropwise addition of ethyl formate (2.2 mL, 27.11 mmol) in THF (2 mL). The mixture was stirred at -78 °C for another hour. LC-MS showed the reaction was complete. The mixture was quenched with a saturated aqueous solution of _NH4Cl and extracted with EA (50 mL × 3). The combined organic phases were washed with brine, dried over _{anhydrous Na2SO4} , filtered, and concentrated to dryness. The residue was purified by silica gel column chromatography (PE:EA = 10:1, V/V) to give the desired product (1.7 g, yield: 48%). ¹ H NMR (400MHz, DMSO) δ 10.11 (s, 1H), 7.91 (d, J = 4.0Hz, 1H).

Step 2. (2,6-Dichloro-3-fluoropyridin-4-yl)methanol (34-4)

At 0 °C, _NaBH₄ (590 mg, 17.53 mmol) was added fractionally to a solution of 2,6-dichloro-3-fluoroisocyanuric acid (1.7 g, 8.76 mmol) in THF (30 mL). After addition, the mixture was stirred at 0 °C for 1 hour. _LC -MS showed that the reaction was complete. The reaction mixture was quenched with saturated _NH₄Cl aqueous solution and extracted with EA (40 mL × 3). The combined organic phases were washed with brine, dried over anhydrous _Na₂SO₄ , filtered, and concentrated to dryness. The residue was purified by silica gel column chromatography (PE:EA = 5:1, V/V) to give the desired product (1.62 g, yield: 95%). LC/MS (ESI): m/z 196 [M+H] ^⁺ .

Step 3.2, 6-Dichloro-4-(chloromethyl)-3-fluoropyridine (34-5)

SOCl₂ ₍ 1.2 mL, 16.33 mmol) was added dropwise to a solution of (2,6-dichloro-3-fluoropyridin-4-yl)methanol (1.6 g, 8.16 mmol) and DMF (0.05 mL, 0.68 mmol) in DCM (30 mL) at 0 °C. The mixture was stirred at room temperature for 16 hours. LC-MS showed that the reaction was complete. The reaction mixture was concentrated to give the desired product (1.7 g, yield: 97%). LC/MS (ESI) m/z: 214 [M+H] ^⁺ .

Step 4.2, 6-Dichloro-3-fluoro-4-((methanesulfonyl)methyl)pyridine (34-6)

At 0 °C, _CH₃SO₂Na ( _1.21 g, 11.89 mmol) was added fractionally to a solution of 2,6-dichloro-4-(chloromethyl)-3-fluoropyridine (1.7 g, 7.93 mmol) in DMF (30 mL). The mixture was stirred at room temperature for 4 hours. _LC -MS showed that the reaction was complete. The reaction mixture was poured into _H₂O (20 mL) and extracted with EA (30 mL × 3). The combined organic phases were washed with brine, dried over anhydrous _Na₂SO₄ , filtered, and concentrated to dryness. The residue was purified by silica gel rapid column chromatography (PE:EA = 3:1, V/V) to give the desired product (1.76 g, yield: 86%). LC/MS (ESI): m/z 258 [M+H] ^⁺ .

Step 5.2, 6-Dichloro-3-fluoro-4-(1-(methanesulfonyl)cyclopropyl)pyridine (34-7)

To a solution of 2,6-dichloro-3-fluoro-4-((methanesulfonyl)methyl)pyridine (1.76 g, 6.82 mmol), 1,2-dibromoethane (1.5 mL, 17.05 mmol), and TBAB (440 mg, 1.36 mmol) in toluene (60 mL), NaOH (10 M in _H₂O , 6.82 mL, 68.19 mmol) was added. The mixture was stirred at 60 °C for 3 hours. LC-MS showed that the reaction was complete. The reaction mixture was poured into _H₂O (30 mL) and extracted with EA (30 mL × ₃ ). The combined organic phases were washed with brine, dried over anhydrous _Na₂SO₄ , filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 2:1, V/V) to give the desired product (1.6 g, yield: 83%). LC/MS (ESI): m/z 284 [M+H] ^⁺ .

Step 6. 6-Chloro-3-fluoro-4-(1-(methanesulfonyl)cyclopropyl)-N-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-5-yl)pyridine-2-amine (34-9)

Pd₂(dba)₃ (193 mg, 0.21 mmol) and C₂S₂CO₃ (1.38 g, 4.22 mmol) were added to a solution of 2,6-dichloro-3-fluoro-4-(1-(methanesulfonyl)cyclopropyl)pyridine (600 mg, 2.11 mmol), ₁ -((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole- ₅ -amine (450 mg, 2.11 mmol), _XantPhos (244 mg, 0.42 mmol) in _dioxane (15 mL). The mixture was stirred for 6 hours at 100 °C under a _nitrogen atmosphere. _LC -MS showed that the reaction was complete. The reaction mixture was diluted with EA (40 mL), washed with water and brine, dried over anhydrous _Na₂SO₄ , filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 1:1, V/V) to obtain the desired product (715 mg, yield: 73.5%). LC/MS (ESI): m/z 461 [M+H] ⁺ .

Step 7. (R)-3-fluoro-6-(3-methylmorpholino)-4-(1-(methanesulfonyl)cyclopropyl)-N-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-5-yl)pyridine-2-amine (34-11)

Pd₂(dba)₃ (127 mg, 0.14 mmol) and C₂S₂CO₃ (1.36 g, 4.16 mmol) were added to a solution of 6-chloro-3-fluoro-4-(1-(methanesulfonyl)cyclopropyl)-N-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-5-yl)pyridin-2-amine (640 mg, 1.39 mmol), (3R) _{-3 -} methylmorpholine (281 mg, 2.78 mmol), and _RuPhos (130 mg, 0.28 mmol) in _dioxane (15 mL). The mixture was stirred for 16 hours at 100 °C under a _nitrogen atmosphere. LC-MS showed that the reaction was complete. The reaction mixture was diluted with EA (40 mL), washed with water and brine, dried over anhydrous _{Na₂SO₄ ,} filtered, and concentrated. The residue was purified by silica gel rapid column chromatography (DCM:MeOH = 50:1, V/V) to obtain the desired product (450 mg, yield: 62%). LC/MS (ESI): m/z 527 [M+H] ⁺ .

Step 8. (R)-3-fluoro-6-(3-methylmorpholino)-4-(1-(methanesulfonyl)cyclopropyl)-N-(1H-pyrazol-5-yl)pyridin-2-amine (34)

A mixture of (R)-3-fluoro-6-(3-methylmorpholino)-4-(1-(methanesulfonyl)cyclopropyl)-N-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-5-yl)pyridine-2-amine (450 mg, 0.86 mmol) in HCl solution (4 M in dioxane, 5 mL) was stirred for 1.5 h at 60 °C. LC-MS showed the reaction was complete. The mixture was concentrated under vacuum. The residue was purified by silica gel rapid column chromatography (DCM:MeOH = 30:1, V/V) to give a pale yellow oil, which could be further purified by preparative HPLC (C ₁₈ , 10-95% MeOH in H ₂ O, containing 0.1% HCOOH) to give the desired product (45 mg, yield: 13%). LC/MS(ESI):m/z 396[M+H] ⁺ . ¹ H NMR (400MHz, DMSO) δ12.17(s,1H),8.80(s,1H),7.58(s,1H),6.47(s,1H),6. 11(s,1H),4.22–4.15(m,1H),3.93(dd,J＝11.2,3.1Hz,1H),3.74–3.66(m,2H) ,3.61(dd,J＝11.3,2.7Hz,1H),3.48–3.46(m,1H),3.04(dd,J＝12.5,3.7Hz,1 H), 2.98 (s, 3H), 1.70–1.62 (m, 2H), 1.37–1.30 (m, 2H), 1.11 (d, J = 6.6Hz, 3H).

Example 29

Step 1.2-{2-chloro-6-[(3R)-3-methylmorpholin-4-yl]pyridin-4-yl}acetonitrile (35-1)

NaCN (219 mg, 4.47 mmol) was added fractionally to a solution of (3R)-4-[6-chloro-4-(chloromethyl)pyridin-2-yl]-3-methylmorpholine (778 mg, 2.98 mmol) in DMSO (15 mL). The mixture was stirred overnight at ambient temperature. _LC -MS showed that the reaction was complete. The mixture was diluted with EA (50 mL), washed with water and brine, dried over anhydrous _Na₂SO₄ , filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 3:1, V/V) to give the desired product (253 mg, yield: 34%). LC/MS ESI (m/z): 252 [M+H] ^⁺ .

Step 2.1-{2-chloro-6-[(3R)-3-methylmorpholin-4-yl]pyridin-4-yl}cyclopropane-1-carboxylonitrile (35-2)

At 60 °C under a nitrogen atmosphere, a mixture of 2-{2-chloro-6-[(3R)-3-methylmorpholin-4-yl]pyridin-4-yl}acetonitrile (80 mg, 0.32 mmol), 1,2-dibromoethane (0.06 mL, 0.64 mmol), KOH (500 mg, 8.91 mmol), and TBAB (21 mg, 0.06 mmol) in 2-methyltetrahydrofuran ( ₂ mL) and H₂O (0.5 mL) was stirred overnight. LC-MS showed the reaction was complete. The mixture was diluted with EA (50 mL), washed with water and brine, dried over anhydrous _{Na₂SO₄ ,} filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 2:1, V/V) to give the desired product (64 mg, yield: 73%). LC/MS ESI (m/z): 278 [M+H] ^⁺ .

Step 3.5-{[4-(1-cyanocyclopropyl)-6-[(3R)-3-methylmorpholin-4-yl]pyridin-2-yl]amino}-3-methyl-1H-pyrazole-1-carboxylic acid tert-butyl ester (35-3)

A mixture of 1-{2-chloro-6-[(3R)-3-methylmorpholin-4-yl]pyridin-4-yl}cyclopropane-1-carboxylonitrile (64 mg, 0.23 mmol), tert-butyl 5-amino-3-methyl-1H-pyrazole-1-carboxylate (68 mg, 0.35 mmol), BrettPhos-Pd-G3 (20 mg, 0.02 mmol), and _Cs₂CO₃ (225 mg, 0.69 mmol) in _dioxane (6 mL) was stirred overnight at 100 °C under a nitrogen atmosphere. LC-MS showed the reaction was complete. The mixture was diluted with EA (50 mL), washed with water and brine, _dried over anhydrous _Na₂SO₄ , filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 1:1, V/V) to give the desired product (62 mg, yield: 61%). LC/MS ESI(m/z):439[M+H] ⁺ .

Step 4.1-{2-[(3-methyl-1H-pyrazol-5-yl)amino]-6-[(3R)-3-methylmorpholin-4-yl]pyridin-4-yl}cyclopropane-1-carboxylonitrile (35)

At ambient temperature, a solution of 5-{[4-(1-cyanocyclopropyl)-6-[(3R)-3-methylmorpholin-4-yl]pyridin-2-yl]amino}-3-methyl-1H-pyrazole-1-carboxylic acid tert-butyl ester (62 mg, 0.14 mmol) in HCl solution (4 M in dioxane, 4 mL) was stirred overnight. LC-MS showed the reaction was complete. The mixture was concentrated to dryness under reduced pressure. The residue was purified by preparative HPLC ( _C18 , 10-95% MeOH in _H2O containing 0.1% HCOOH) to give the desired product (15.4 mg, yield: 32%). LC/MS ESI (m/z): 339 [M+H] ⁺ . ¹ H NMR(400MHz,DMSO)δ11.72(br,1H),8.81(s,1H),6.57(s,1H),5.99(s,1H),5.77(s,1H),4 .30(d,J＝6.5Hz,1H),3.92(dd,J＝11.2,3.2Hz,1H),3.76(d,J＝12.7Hz,1H),3.71(d,J＝11.2 Hz,1H),3.60(dd,J＝11.2,2.8Hz,1H),3.47–3.44(m,1H),3.03(dd,J＝12.6,9.0Hz,1H),2. 17 (s, 3H), 1.71 (dd, J = 7.5, 4.4Hz, 2H), 1.50 (dd, J = 7.7, 4.4Hz, 2H), 1.11 (d, J = 6.6Hz, 3H).

Example 30

Step 1. (R)-N-(3-methyl-1H-pyrazol-5-yl)-6-(3-methylmorpholino)-4-(4-(methanesulfonyl)tetrahydro-2H-pyran-4-yl)pyridin-2-amine (36)

BrettPhos-Pd-G3 (24 mg, 0.027 mmol) was added to a solution of (R)-4-(6-chloro-4-(4-(methanesulfonyl)tetrahydro-2H-pyran-4-yl)pyridin-2-yl)-3-methylmorpholine (100 mg, 0.27 mmol), tert-butyl 5-amino- ₃ -methyl-1H-pyrazol- ₁ -carboxylate (105 mg, 0.53 mmol), and Cs₂CO₃ (261 mg, 0.80 mmol) in dioxane (8 mL). The mixture was stirred for 16 hours at 100 °C under a _nitrogen atmosphere. LC-MS showed that the reaction was complete. The reaction mixture was diluted with EA (60 mL), washed with water and brine, dried over anhydrous _{Na₂SO₄ ,} filtered, and concentrated to dryness. The residue was purified by preparative HPLC ( _C18 , 10-95% MeOH in _H2O , containing 0.1% HCOOH) to obtain the desired product (12 mg, yield: 10%). LC/MS (ESI): m/z 436 [M+H] ⁺ . ¹ H NMR (400MHz, DMSO) δ 11.69 (s, 1H), 8.82 (s, 1H), 6.70 (s, 1H), 6.17 (s, 1H), 6.05 (s, 1H), 4.31–4.23 (m, 1H), 3.96–3.83 (m, 4H), 3.72 (d, J = 11.2 Hz, 1H), 3.64 (dd, J = 11.2, 2 .7Hz,1H),3.49(td,J＝11.7,2.8Hz,1H),3.26–3.18(m,2H),3.05(td,J＝12.7,3.6Hz ,1H),2.71(s,3H),2.43(d,J＝13.7Hz,2H),2.24–2.16(m,5H),1.12(d,J＝6.6Hz,3H).

Example 31

Step 1.1-{2-chloro-6-[(3R)-3-methylmorpholin-4-yl]pyridin-4-yl}cyclopentane-1-carboxylonitrile (37-1)

At 70 °C, a mixture of 2-{2-chloro-6-[(3R)-3-methylmorpholin-4-yl]pyridin-4-yl}acetonitrile (210 mg, 0.83 mmol), 1,4-dibromobutane (1 mL, 8.34 mmol), KOH (3 g, 53.47 mmol), and TBAB (54 mg, 0.17 mmol) in a co-solvent of 2-methyltetrahydrofuran (15 mL) and _H₂O (3 mL) was stirred overnight. LC-MS showed the reaction was complete. The mixture was diluted with EA (50 mL), washed with water and brine, _dried over anhydrous _Na₂SO₄ , filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 3:1, V/V) to give the desired product (192 mg, yield: 75%). LC/MS ESI (m/z): 306 [M+H] ^⁺ .

Step 2.5-{[4-(1-cyanocyclopentyl)-6-[(3R)-3-methylmorpholin-4-yl]pyridin-2-yl]amino}-1H-pyrazole-1-carboxylic acid tert-butyl ester (37-2)

BrettPhos-Pd-G3 (29.6 mg, 0.03 mmol) and Cs₂CO₃ (319.6 mg, 0.98 mmol) were added to a solution of 1-{2-chloro-6-[(3R)-3-methylmorpholin-4-yl]pyridin-4-yl}cyclopentane-1-carboxynitrile (100 mg, 0.33 mmol) and tert-butyl _{5-amino-1H-pyrazol-1 -} carboxylate (90 mg, 0.49 mmol) in dioxane (10 mL). The mixture was stirred overnight at 100 °C under a nitrogen atmosphere. _LC -MS showed that the reaction was complete. The mixture was diluted with EA (50 mL), washed with water and brine, dried over anhydrous _Na₂SO₄ , filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 1:1, V/V) to give the desired product (104 mg, yield: 70%). LC/MS ESI(m/z):453[M+H] ⁺ .

Step 3.1-{2-[(3R)-3-methylmorpholin-4-yl]-6-[(1H-pyrazol-5-yl)amino]pyridin-4-yl}cyclopentane-1-carboxylonitrile (37)

At ambient temperature, a mixture of 5-{[4-(1-cyanocyclopentyl)-6-[(3R)-3-methylmorpholin-4-yl]pyridin-2-yl]amino}-1H-pyrazole-1-carboxylic acid tert-butyl ester ( ₁₀₄ mg, 0.23 mmol) in HCl solution (4 M in dioxane, 4 mL) was stirred overnight. LC-MS showed the reaction was complete. The mixture was concentrated to dryness under reduced pressure. The residue was purified by preparative HPLC (C18, 10-95% MeOH in _H2O containing 0.1% HCOOH) to give the desired product (42.5 mg, yield: 52%). LC/MS ESI (m/z): 353 [M+H] ⁺ . ¹ H NMR (400MHz, DMSO) δ12.14(s,1H),9.01(s,1H),7.54(d,J＝2.2Hz,1H),6.59(s,1H),6.30(d,J＝2 .2Hz,1H),6.07(d,J＝0.9Hz,1H),4.35–4.29(m,1H),3.93(dd,J＝11.2,3.3Hz,1H),3.79(d,J＝13. 0Hz,1H),3.73(d,J＝11.2Hz,1H),3.62(dd,J＝11.3,2.8Hz,1H),3.48(dd,J＝11.8,2.9Hz,1H),3. 09–3.02(m,1H),2.34–2.28(m,2H),2.10–2.03(m,2H),1.90–1.85(m,4H),1.13(d,J＝6.6Hz,3H).

Example 32

Step 1.1-{2-chloro-6-[(3R)-3-methylmorpholin-4-yl]pyridin-4-yl}cyclohexane-1-carboxylonitrile (38-1)

At 70 °C, a mixture of 2-{2-chloro-6-[(3R)-3-methylmorpholin-4-yl]pyridin-4-yl}acetonitrile (210 mg, 0.83 mmol), 1,5-dibromopentane (1.2 mL, 8.34 mmol), KOH (3 g, 53.47 mmol), and TBAB (54 mg, 0.17 mmol) in a cosolvent of 2-methyltetrahydrofuran (15 mL) and _H₂O (3 mL) was stirred overnight. _LC -MS showed the reaction was complete. The mixture was diluted with EA (50 mL), washed with water and brine, dried over anhydrous _Na₂SO₄ , filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 3:1, V/V) to give the desired product (194 mg, yield: 73%). LC/MS ESI (m/z): 320 [M+H] ^⁺ .

Step 2.5-{[4-(1-cyanocyclohexyl)-6-[(3R)-3-methylmorpholin-4-yl]pyridin-2-yl]amino}-1H-pyrazole-1-carboxylic acid tert-butyl ester (38-2)

BrettPhos-Pd-G3 (28 mg, 0.03 mmol) and Cs₂CO₃ (306 mg, 0.94 mmol) were added to a solution of 1-{2-chloro-6-[(3R)-3-methylmorpholin-4-yl]pyridin-4-yl}cyclohexane-1-carboxynitrile (100 mg, 0.31 mmol) and tert-butyl 5-amino-1H-pyrazol _{-1 -} carboxylate (86 mg, 0.47 mmol) in dioxane (10 mL). The mixture was stirred overnight at 100 °C under a nitrogen atmosphere. _LC -MS showed that the reaction was complete. The mixture was diluted with EA (50 mL), washed with water and brine, dried over anhydrous _Na₂SO₄ , filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 1:1, V/V) to give the desired product (102 mg, yield: 70%). LC/MS ESI(m/z):467[M+H] ⁺ .

Step 3.1-{2-[(3R)-3-methylmorpholin-4-yl]-6-[(1H-pyrazol-5-yl)amino]pyridin-4-yl}cyclohexane-1-carboxylonitrile (38)

At ambient temperature, a mixture of 5-{[4-(1-cyanocyclohexyl)-6-[(3R)-3-methylmorpholin-4-yl]pyridin-2-yl]amino}-1H-pyrazole-1-carboxylic acid tert-butyl ester (102 mg, 0.22 mmol) in HCl solution (4 M in dioxane, 4 mL) was stirred overnight. LC-MS showed the reaction was complete. The mixture was concentrated to dryness under reduced pressure. The residue was purified by preparative HPLC ( _C18 , 10-95% MeOH in _H2O containing 0.1% HCOOH) to give the desired product (36.6 mg, yield: 46%). LC/MS ESI (m/z): 367 [M+H] ⁺ . ¹ H NMR (400MHz, DMSO) δ12.17(s,1H),9.00(s,1H),7.53(d,J＝2.2Hz,1H),6.63(s,1H),6.29(d,J＝2.1Hz,1H),6.11(d ,J＝0.8Hz,1H),4.32(d,J＝6.6Hz,1H),3.93(dd,J＝11.2,3.3Hz,1H),3.80(d,J＝11.3Hz,1H),3.73(d,J＝11.1Hz,1H) ,3.62(dd,J＝11.3,2.8Hz,1H),3.47(td,J＝11.8,3.0Hz,1H),3.05(td,J＝12.6,3.7Hz,1H),2.00(d,J＝12.4Hz,2H), 1.81(dd,J＝16.4,6.6Hz,4H), 1.73(d,J＝13.6Hz,1H), 1.66–1.56(m,2H), 1.34–1.24(m,1H), 1.12(d,J＝6.6Hz,3H).

Example 33

Step 1. (R)-4-(2-chloro-6-(3-methylmorpholino)pyridin-4-yl)tetrahydro-2H-pyran-4-carboxynitrile (39-1)

At 60 °C, a mixture of 2-{2-chloro-6-[(3R)-3-methylmorpholin-4-yl]pyridin-4-yl}acetonitrile (180 mg, 0.72 mmol), 1-bromo-2-(2-bromoethoxy)ethane (660 mg, 2.85 mmol), TBAB (46 mg, 0.14 mmol), and NaOH (10.0 M in _H₂O , _14.0 mmol, _1.4 mL) in toluene (10 mL) was stirred for 2 hours. LC-MS showed the reaction was complete. The reaction mixture was diluted with EA (40 mL), washed with water and brine, dried over anhydrous Na₂SO₄, filtered, and concentrated under vacuum. The residue was purified by silica gel column chromatography (PE:EA = 2:1, V/V) to give the desired product (157 mg, yield: 68%). LC/MS (ESI): m/z 322 [M+H] ^⁺ .

Step 2. (R)-4-(2-((1H-pyrazol-5-yl)amino)-6-(3-methylmorpholino)pyridin-4-yl)tetrahydro-2H-pyran-4-carboxynitrile (39)

A mixture of 4-{2-chloro- ₆ -[(3R)-3-methylmorpholin-4-yl]pyridin-4-yl}tetrahydropyran-4-carboxynitrile (80 mg, 0.25 mmol), 1H-pyrazol-5-amine (41 mg, 0.49 mmol), BrettPhos-Pd-G3 (22 mg, 0.02 mmol), and _Cs₂CO₃ (244 _mg , 0.75 mmol) in dioxane (4 mL) was stirred for 16 hours at ₁₁₀ °C under a N₂ atmosphere. LC-MS showed the reaction was complete. The reaction mixture was diluted with EA (40 mL), washed with water and brine, dried over anhydrous _Na₂SO₄ , filtered, and concentrated. The residue was purified by preparative HPLC ( _C₁₈ , 10-95% MeOH in _H₂O containing 0.1% HCOOH) to give the desired product (37 mg, yield: 40%). LC/MS(ESI):m/z 369[M+H] ⁺ . ¹ H NMR (400MHz, DMSO) δ12.06(s,1H),9.04(s,1H),7.54(d,J＝1.9Hz,1H),6.64(s,1H),6.29( s,1H),6.13(s,1H),4.34(d,J＝6.9Hz,1H),4.00(dd,J＝11.2,3.0Hz,2H),3.93(dd,J＝11.3, 3.3Hz,1H),3.82(d,J＝12.8Hz,1H),3.73(d,J＝11.2Hz,1H),3.68–3.59(m,3H),3.47(td,J ＝11.8,2.9Hz,1H),3.06(td,J＝12.6,3.7Hz,1H),2.13–1.95(m,4H),1.13(d,J＝6.6Hz,3H).

Example 34

Step 1. (R)-1-(2-chloro-6-(3-methylmorpholino)pyrimidin-4-yl)cyclopentane-1-carboxylonitrile (40-1)

Add an aqueous solution of KOH (10 M, 1.58 mL, 15.8 mmol) to a solution of (R)-2-(2-chloro-6-(3-methylmorpholino)pyrimidin-4-yl)acetonitrile (200 mg, 0.79 mmol), 1,4-dibromobutane (0.95 mL, 7.91 mmol), and TBAB (26 mg, 0.08 mmol) in 15 mL of 2-MeTHF. Stir the mixture overnight at 70 °C. LC-MS showed the reaction was complete. Dilute the reaction mixture with EA (60 mL), wash with water and brine, _dry over anhydrous _Na₂SO₄ , filter, and concentrate to dryness. Purify the residue by silica gel column chromatography (PE:EA = 10:1, V/V) to give the desired product (220 mg, yield: 91%). LC/MS (ESI): m/z 307 [M+H] ^⁺ .

Step 2. (R)-1-(2-((1H-pyrazol-5-yl)amino)-6-(3-methylmorpholino)pyrimidin-4-yl)cyclopentane-1-carboxynitrile (40)

BrettPhos-Pd- _G3 (29 mg, 0.03 mmol) was added to a suspension of (R)-1-(2-chloro-6-(3-methylmorpholino)pyrimidin-4-yl)cyclopentane-1-carboxynitrile (100 mg, 0.33 mmol), 1H-pyrazol _-5- amine (41 mg, 0.49 mmol), and Cs₂CO₃ (319 mg, 0.98 mmol) in dioxane (8 mL). The mixture was stirred at 100 °C for 16 h. LC-MS showed that the reaction was complete. The reaction mixture was diluted with DCM (50 mL), washed with water and brine, dried over _{anhydrous Na₂SO₄} , filtered, and concentrated to dryness. The residue was purified by preparative HPLC ( _C₁₈ , 10-95% MeOH in _H₂O containing 0.1% HCOOH) to give the desired product (30 mg, yield: 26%). LC/MS(ESI):m/z 354.4[M+H] ⁺ . ¹ H NMR(400MHz,DMSO)δ12.09(s,1H),9.19(s,1H),7.52(s,1H),6.49(s,1H),6.28(s ,1H),4.40(d,J＝5.4Hz,1H),4.00(d,J＝12.9Hz,1H),3.93(dd,J＝11.3,3.3Hz,1H), 3.72(d,J＝11.4Hz,1H),3.59(dd,J＝11.4,2.9Hz,1H),3.46–3.40(m,1H),3.14(td ,J＝12.9,3.7Hz,1H),2.31–2.22(m,4H),1.89–1.82(m,4H),1.19(d,J＝6.7Hz,3H).

Example 35

Step 1. (R)-1-(2-chloro-6-(3-methylmorpholino)pyrimidin-4-yl)cyclohexane-1-carboxylonitrile (41-1)

A suspension of (R)-2-(2-chloro-6-(3-methylmorpholino)pyrimidin-4-yl)acetonitrile (200 mg, 0.79 mmol), 1,5-dibromopentane (1.08 mL, 7.91 mmol), and TBAB (26 mg, 0.08 mmol) in 2-MeTHF (15 mL) was added to an aqueous solution of KOH (10 M, 1.58 mL, 15.8 mmol). The mixture was stirred at 70 °C for 16 hours. LC-MS showed that the reaction was complete. The reaction mixture was diluted with EA (60 mL), washed with water and brine, dried over anhydrous _{Na₂SO₄ ,} filtered, and concentrated to dryness. The residue was purified by silica gel column chromatography (PE:EA = 10:1, V/V) to give the desired product (210 mg, yield: 83%). LC/MS (ESI): m/z 321 [M+H] ^⁺ .

Step 2. (R)-1-(2-((1H-pyrazol-5-yl)amino)-6-(3-methylmorpholino)pyrimidin-4-yl)cyclohexane-1-carboxynitrile (41)

BrettPhos-Pd- _G3 (28 mg, 0.03 mmol) was added to a suspension of (R)-1-(2-chloro-6-(3-methylmorpholino)pyrimidin-4-yl)cyclohexane-1-carboxynitrile (100 mg, 0.31 mmol), 1H-pyrazol- _5- amine (39 mg, 0.47 mmol), and Cs₂CO₃ (305 mg, 0.94 mmol) in dioxane (8 mL). The mixture was stirred at 100 °C for 16 h. LC-MS showed that the reaction was complete. The reaction mixture was diluted with EA (50 mL), washed with water and brine, dried over _{anhydrous Na₂SO₄} , filtered, and concentrated to dryness. The residue was purified by preparative HPLC ( _C₁₈ , 10-95% MeOH in _H₂O containing 0.1% HCOOH) to give the desired product (20 mg, yield: 17.5%). LC/MS(ESI):m/z 368.5[M+H] ⁺ . ¹ H NMR(400MHz,DMSO)δ12.09(s,1H),9.18(s,1H),7.51(s,1H),6.52(s,1H),6.26(s,1H),4.46–4.34(m,1 H),4.01(d,J＝13.3Hz,1H),3.93(dd,J＝11.4,3.3Hz,1H),3.72(d,J＝11.4Hz,1H),3.58(dd,J＝11.4,3.0 Hz,1H),3.43(td,J＝11.9,2.9Hz,1H),3.14(td,J＝12.8,3.6Hz,1H),2.09–2.02(m,2H),1.95–1.86(m,2 H),1.85–1.78(m,2H),1.77–1.70(m,1H),1.65–1.53(m,2H),1.32–1.22(m,1H),1.19(d,J＝6.7Hz,3H).

Example 36

Step 1. (R)-4-(2-chloro-6-(3-methylmorpholino)pyrimidin-4-yl)tetrahydro-2H-pyran-4-carboxynitrile (42-1)

Add 1-bromo-2-(2-bromoethoxy)ethane (367 mg, 1.58 mmol), sodium hydroxide (10 M in _H₂O , 0.79 mL, 7.91 mmol), and TBAB (52 mg, 0.16 mmol) to a solution of (R)-2-(2-chloro-6-(3-methylmorpholino)pyrimidin-4-yl)acetonitrile (200 mg, 0.79 mmol) in toluene (10 mL). Stir the reaction overnight at 60 °C. LC-MS showed the reaction was complete. Dilute the mixture with EA (60 mL), wash with water and brine, _dry over anhydrous _Na₂SO₄ , filter, and concentrate. Purify the residue by silica gel rapid chromatography (PE:EA = 3:1, V/V) to give the desired product (180 mg, yield: 70%). LC/MS (ESI): m/z 323 [M+H] ^⁺ .

Step 2. (R)-4-(2-((1H-pyrazol-5-yl)amino)-6-(3-methylmorpholino)pyrimidin-4-yl)tetrahydro-2H-pyran-4-carboxynitrile (42)

To a solution of (R)-4-(2-chloro-6-(3-methylmorpholino)pyrimidin-4-yl)tetrahydro-2H-pyran-4-carboxynitrile (80 mg, 0.25 mmol) in dioxane (8 mL), 1H-pyrazol-5 _- amine (31 mg, 0.37 mmol), _Cs₂CO₃ (162 mg, 0.50 mmol), and BrettPhos-Pd- _G₃ (45 mg, 0.05 mmol) were added. The mixture was stirred overnight at 100 °C under a nitrogen atmosphere. LC-MS showed that the reaction was complete. The mixture was diluted with EA (60 mL), washed with water and brine, dried over anhydrous _Na₂SO₄ , filtered, and concentrated. The residue was purified by preparative HPLC (C18, 10-95% MeOH in _H₂O containing 0.1% HCOOH) to give the desired product (15 mg, yield: 16%). LC/MS(ESI):m/z 370[M+H] ⁺ . ¹ HNMR(400MHz,DMSO)δ12.10(s,1H),9.14(s,1H),7.54(s,1H),6.60(s,1H),6.29(s,1H), 4.43(s,1H),4.07–3.96(m,3H),3.93(dd,J＝11.4,3.4Hz,1H),3.72(d,J＝11.4Hz,1H),3. 63(dd,J＝12.0,10.3Hz,2H),3.57(d,J＝3.0Hz,1H),3.43(td,J＝11.9,2.9Hz,1H),3.15(t d,J＝12.9,3.6Hz,1H),2.20–2.12(m,2H),2.03(d,J＝12.3Hz,2H),1.20(d,J＝6.7Hz,3H).

Example 37

Step 1.1 - (2,6-dichloropyridin-4-yl)cyclohexyl-1-ol (43-3)

At -78 °C, n-BuLi solution (2.5 M in THF, 0.74 mL, 1.85 mmol) was added dropwise to a solution of 4-bromo-2,6-dichloropyridine (300 mg, 1.32 mmol) and cyclohexanone (156 mg, 1.59 mmol) in THF (8 mL). The mixture was stirred at -78 °C for 1 hour. LC-MS showed that the reaction was complete. The mixture was quenched with saturated _NH4Cl aqueous solution and extracted three times with EA (30 mL × 3). The combined organic phases were washed with brine, dried over _{anhydrous Na2SO4} , filtered, and concentrated to dryness. The residue was purified by silica gel column chromatography (PE:EA = 10:1, V/V) to give the desired product (252 mg, yield: 77%). LC/MS (ESI): m/z 246 [M+H] ⁺ .

Step 2. (R)-1-(2-chloro-6-(3-methylmorpholino)pyridin-4-yl)cyclohexyl-1-ol (43-5)

(3R)-3-methylmorpholine (308 mg, 3.05 mmol) was added to a solution of 1-(2,6-dichloropyridin-4-yl)cyclohexyl-1-ol (250 mg, 1.02 mmol) in NMP (5 mL). The mixture was stirred at 150 °C for 16 h. LC-MS showed that the reaction was complete. The reaction mixture was diluted with EA (60 mL), washed with water and brine, dried over _{anhydrous Na₂SO₄} , filtered, and concentrated to dryness. The residue was purified by silica gel column chromatography (PE:EA = 5:1, V/V) to give the desired product (42 mg, yield: 13.3%). LC/MS (ESI): m/z 311 [M+H] ^⁺ .

Step 3. (R)-1-(2-((1H-pyrazol-5-yl)amino)-6-(3-methylmorpholino)pyridin-4-yl)cyclohexyl-1-ol (43)

BrettPhos-Pd- _G3 (12 mg, 0.01 mmol) was added to a suspension of (R)-1-(2-chloro-6-(3-methylmorpholino)pyridin-4-yl)cyclohexyl-1-ol (42 mg, 0.135 mmol), 1H-pyrazol- _5- amine (23 mg, 0.270 mmol), and Cs₂CO₃ (132 mg, 0.405 mmol) in dioxane (6 _mL ). The mixture was stirred for 16 hours at 100 °C under a _nitrogen atmosphere. LC-MS showed that the reaction was complete. The reaction mixture was diluted with EA (40 mL), washed with water and brine, dried over anhydrous _Na₂SO₄ , filtered, and concentrated to dryness. The residue was purified by preparative HPLC ( _C₁₈ , 10-95% MeOH in _H₂O containing 0.1% HCOOH) to give the desired product (10 mg, yield: 21%). LC/MS(ESI):m/z 358[M+H] ⁺ . ¹ H NMR (400MHz, DMSO) δ12.17(s,1H),8.74(s,1H),7.50(d,J＝2.2Hz,1H),6.54(s,1H),6.28(d, J＝2.0Hz,1H),6.15(s,1H),4.60(s,1H),4.32–4.23(m,1H),3.93(dd,J＝11.1,3.1Hz,1H),3.7 7–3.68(m,2H),3.62(dd,J＝11.2,2.8Hz,1H),3.47(td,J＝11.7,2.9Hz,1H),3.03(td,J＝12.5 ,3.6Hz,1H),1.73–1.53(m,7H),1.51–1.43(m,2H),1.27–1.17(m,1H),1.11(d,J＝6.6Hz,3H).

Example 38

Step 1. 4-Bromo-2,6-Dichloropyrimidine (44-3)

At -60 °C, a solution of lithium tetramethylpiperidin (1.0 M in THF, 8.0 mL, 8.05 mmol) was added dropwise to a solution of 2,4-dichloropyrimidine (1 g, 6.71 mmol) in THF (50 mL). The mixture was stirred at -60 °C for 1 hour, followed by the dropwise addition of a solution of 1,2-dibromo-1,1,2,2-tetrachloroethane (3.28 g, 10.07 mmol) in THF (5 mL). The resulting mixture was stirred at -60 °C for another 2 hours. LC-MS showed the reaction was complete. The reaction mixture was quenched with a saturated aqueous solution of _NH₄Cl and extracted with EA (60 mL). The organic layer was separated, washed with brine, dried over _{anhydrous Na₂SO₄} , filtered, and concentrated. The residue was purified by silica gel rapid chromatography (PE:EA = 50:1, V/V) to give the desired product (600 mg, yield: 39%). LC/MS(ESI): m/z 228[M+H] ⁺ .

Step 2.1 - (2,6-Dichloropyrimidin-4-yl)cyclohexyl-1-ol (44-5)

At -60 °C, n-butyllithium (2.5 M in THF, 1.5 mL, 3.69 mmol) was added dropwise to a solution of 4-bromo-2,6-dichloropyrimidine (600 mg, 2.63 mmol) and cyclohexanone (0.32 mL, 3.16 mmol) in 15 mL of THF. The mixture was stirred at -60 °C for 30 min. LC-MS showed the reaction was complete. The reaction mixture was quenched with saturated _NH4Cl aqueous solution and extracted with EA (60 mL). The organic layer was separated, washed with brine, dried over _{anhydrous Na2SO4} , filtered, and concentrated. The residue was purified by silica gel rapid chromatography (PE:EA = 20:1, V/V) to give the desired product (200 mg, yield: 30%). LC/MS (ESI): m/z 248 [M+H] ⁺ .

Step 3. (R)-1-(2-chloro-6-(3-methylmorpholino)pyrimidin-4-yl)cyclohexyl-1-ol (44-7)

(R)-3-methylmorpholine (246 mg, 2.43 mmol) was added to a solution of 1-(2,6-dichloropyrimidin-4-yl)cyclohexane-1-ol (200 mg, 0.81 mmol) in NMP (5 mL). The mixture was stirred for 1 hour at 120 °C under microwave irradiation. LC-MS showed that the reaction was complete. The mixture was diluted with EA (60 mL), washed with water and brine, _dried over anhydrous _Na₂SO₄ , filtered, and concentrated. The residue was purified by silica gel rapid chromatography (PE:EA = 10:1, V/V) to give the desired product (200 mg, yield: 79%). LC/MS (ESI): m/z 312 [M+H] ^⁺ .

Step 4. (R)-1-(2-((1H-pyrazol-5-yl)amino)-6-(2-methylpiperidin-1-yl)pyrimidin-4-yl)cyclohexyl-1-ol (44)

To a solution of (R)-1-(2-chloro-6-(3-methylmorpholino)pyrimidin-4-yl)cyclohexyl-1-ol (80 mg, 0.26 mmol) in dioxane (2 mL), 1H-pyrazol-5-amine (32 mg, 0.39 mmol), _Cs₂CO₃ (167 mg, 0.51 mmol), and BrettPhos-Pd-G₃ ( ₂₃ mg, 0.03 mmol) were added. The mixture was stirred overnight at 80 °C under a nitrogen atmosphere. _LC -MS showed that the reaction was complete. The mixture was diluted with EA (50 mL), washed with water and brine, dried over anhydrous _Na₂SO₄ , filtered, and concentrated. The residue was purified by preparative HPLC (C18, 10-95% MeOH in _H₂O containing 0.1% HCOOH) to give the desired product (20 mg, yield: 21%). LC/MS(ESI)m/z:317[M+H] ⁺ . ¹ H NMR (400MHz, DMSO) δ8.99 (s, 1H), 7.50 (d, J = 2.0Hz, 1H), 6.41 (d, J = 1.7Hz, 1H), 6.38 (s, 1H), 4. 86(s,1H),4.35(d,J＝4.7Hz,1H),3.94(dd,J＝19.7,8.5Hz,2H),3.73(d,J＝11.3Hz,1H),3.58(d ,J＝8.5Hz,1H),3.43(d,J＝2.9Hz,1H),3.12(dd,J＝12.6,9.2Hz,1H),1.85(dt,J＝12.2,7.2Hz,2 H), 1.69 (dd, J = 24.2, 11.5Hz, 3H), 1.49 (t, J = 13.8Hz, 4H), 1.24 (s, 1H), 1.18 (d, J = 6.7Hz, 3H).

Example 39

Step 1. (R)-4-(6-chloro-4-((methanesulfonyl)methyl)pyridin-2-yl)-3-methylmorpholine (45-1)

A solution of (3R)-4-[6-chloro-4-(chloromethyl)pyridin-2-yl]-3-methylmorpholine ( ₂ g, 7.66 mmol) in DMF (40 mL) was added to a solution of _{Na₂CH₃SO₂} (1.56 g, 15.32 mmol). The mixture was stirred at room temperature for 16 hours. _LC -MS showed that the reaction was complete. The reaction mixture was diluted with EA (100 mL), washed with water and brine, dried over anhydrous _Na₂SO₄ , filtered, and concentrated to dryness. The residue was purified by silica gel column chromatography (PE:EA = 10:1, V/V) to give the desired product (1.6 g, yield: 68%). LC/MS (ESI): m/z 305 [M+H] ^⁺ .

Step 2. (R)-4-(6-chloro-4-(4-(methylsulfonyl)tetrahydro-2H-pyran-4-yl)pyridin-2-yl)-3-methylmorpholine (45-2)

An aqueous solution of NaOH (10 M, 2.62 mmol) was added to a solution of (R)-4-(6-chloro-4-((methanesulfonyl)methyl)pyridin-2-yl)-3-methylmorpholine (800 mg, 2.62 mmol), 1-bromo-2-(2-bromoethoxy)ethane (1.83 g, 7.87 mmol), and TBAB (170 mg, 0.53 mmol) in toluene (26 mL). The mixture was stirred at 60 °C for 16 hours. _LC -MS showed that the reaction was complete. The reaction mixture was diluted with EA (60 mL), washed with water and brine, dried over anhydrous _Na₂SO₄ , filtered, and concentrated to dryness. The residue was purified by silica gel column chromatography (PE:EA = 10:1, V/V) to give the desired product (550 mg, yield: 56%). LC/MS (ESI): m/z 375 [M+H] ^⁺ .

Step 3. (R)-6-(3-methylmorpholino)-4-(4-(methanesulfonyl)tetrahydro-2H-pyran-4-yl)-N-(1H-pyrazol-5-yl)pyridin-2-amine (45)

BrettPhos-Pd-G3 (24 mg, 0.03 mmol) was added to a solution of (R)-4-(6-chloro-4-(4-(methanesulfonyl)tetrahydro-2H-pyran-4-yl)pyridin- ₂ -yl) _-3 -methylmorpholine (100 mg, 0.27 mmol), 1H-pyrazole-5 (44 mg, 0.53 mmol), and Cs₂CO₃ (261 mg, 0.80 mmol) in dioxane (8 _mL ). The mixture was stirred at 100 °C for 5 hours. LC-MS showed that the reaction was complete. The reaction mixture was diluted with EA (60 mL), washed with water and brine, dried over anhydrous _Na₂SO₄ , filtered, and concentrated to dryness. The residue was purified by preparative HPLC ( _C₁₈ , 10-95% MeOH in _H₂O containing 0.1% HCOOH) to give the desired product (37 mg, yield: 33%). LC/MS(ESI):m/z 422[M+H] ⁺ . ¹ H NMR (400MHz, DMSO) δ11.94(s,1H),8.98(s,1H),7.54(d,J＝2.2Hz,1H),6.61(s,1H),6.35( d,J＝2.1Hz,1H),6.18(s,1H),4.32–4.23(m,1H),3.97–3.83(m,4H),3.73(d,J＝11.1Hz,1H ),3.64(dd,J＝11.2,2.8Hz,1H),3.52–3.46(m,1H),3.26–3.20(m,2H),3.06(td,J＝12.7,3 .7Hz,1H),2.71(s,3H),2.44(d,J＝13.7Hz,2H),2.25–2.14(m,2H),1.12(d,J＝6.6Hz,3H).

Example 40

Step 1: (R)-4-(6-chloro-4-(1,4-dimethyl-1H-pyrazol-5-yl)pyridin-2-yl)-3-methylmorpholine (46-2)

To a solution of (R)-4-(6-chloro-4-iodopyridin-2-yl)-3-methylmorpholine (150 mg, 0.44 mmol) and 1,4-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxanepentobor-2-yl)-1H-pyrazole (108.24 _mg , 0.49 mmol) in dioxane (5 mL), _K₂CO₃ (121.44 mg, 0.89 mmol) and Pd(dppf) _Cl₂ (32.20 mg, 0.04 mmol) were added. The mixture was injected twice with _N₂ and stirred at 90 °C for 12 hours. The reaction mixture was diluted with water (10 mL) and extracted with EA (15 mL × 2). The combined organic layers were washed with brine (10 mL), dried over _{anhydrous Na₂SO₄} , filtered, and concentrated under vacuum. The resulting mixture was purified by rapid chromatography using PE/EtOAc (3:1, V/V) elution to give the desired product (110 mg, yield: 80.93%).

Step 2: (R)-5-((4-(1,4-dimethyl-1H-pyrazol-5-yl)-6-(3-methylmorpholino)pyridin-2-yl)amino)-3-methyl-1H-pyrazol-1-carboxylic acid tert-butyl ester (46-3)

To a solution of (R)-4-(6-chloro-4-(1,4-dimethyl-1H-pyrazol-5-yl)pyridin- _2- yl)-3-methylmorpholine (110 mg, 0.36 mmol) and tert-butyl 5-amino-3-methyl-1H-pyrazol-1-carboxylate (84.86 mg, 0.43 mmol) in dioxane (5 mL), _CS₂CO₃ (350.47 mg, 1.08 mmol) and BrettPhos Pd G₃ (32.67 mg, 0.036 mmol) were added. The mixture was injected twice with _N₂ and stirred overnight at 90 °C. The reaction mixture was diluted with water and extracted with EtOAc (10 mL × 2). The combined organic layers were washed with brine (10 mL), dried over _{anhydrous Na₂SO₄} , filtered, and concentrated under vacuum. The resulting mixture was purified by rapid chromatography using PE/EtOAc (2:1, V/V) elution to give the desired product (120 mg, yield: 71.58%).

Step 3: (R)-4-(1,4-dimethyl-1H-pyrazol-5-yl)-N-(3-methyl-1H-pyrazol-5-yl)-6-(3-methylmorpholino)pyridine-2-amine (46)

TFA (1 mL) was added to a mixture of (R)-5-((4-(1,4-dimethyl-1H-pyrazol-5-yl)-6-(3-methylmorpholino)pyridin-2-yl)amino)-1H-pyrazol-1-carboxylic acid tert-butyl ester (50 mg, 0.11 mmol) in DCM (3 mL), and the mixture was stirred at room temperature for 1 hour. The resulting mixture was concentrated under vacuum. A saturated solution of NaHCO3 was added to the mixture until pH = 7-8, and extraction was performed with DCM (10 mL × 2). The combined organic layers were washed with brine (10 mL), dried over _{anhydrous Na₂SO₄} , filtered, and concentrated under vacuum. The residue was purified by preparative TLC (DCM/MeOH = 10/1, V/V) to give the desired product (10 mg, yield: 25.45%). LC/MS(ESI)m/z:368.1[M+H] ⁺ . ¹ HNMR(400MHz,DMSO-d ₆ )δppm 11.69(br s,1H)8.88(br s,1H)7.31(s,1H)6.49(br s,1H)6.05(br s,1H)5.98-6.01(m,1H)4.29(br d,J＝6.16Hz,1H)3.86-3.96(m,2H)3.73(s,3H)3.69-3.72(m,1H)3.61-3.66(m,1H)3.45-3 .52(m,1H)3.07(td,J＝12.58,3.34Hz,1H)2.18(s,3H)1.98(s,3H)1.15(d,J＝6.54Hz,3H).

Example 41

Step 1. (R)-4-(3,5-dimethylisoxazol-4-yl)-N-(3-methyl-1H-pyrazol-5-yl)-6-(3-methylmorpholino)pyridine-2-amine (47)

Cs₂CO₃ (400 mg, 1.23 mmol) and BrettPhos Pd G₃ (45 mg, 0.049 mmol) were added to dioxane (8 mL) containing (R)-4-(6-chloro-4-(3,5-dimethylisoxazol-4-yl)pyridin-2-yl) _{-3- methylmorpholine} (150 mg, 0.49 mmol) and tert-butyl 5-amino-1H-pyrazol-1-carboxylate (116 mg, 0.59 mmol). The mixture was injected twice with _N₂ and stirred overnight at 90 °C. The reaction mixture was diluted with water and extracted with EtOAc (30 mL × 2). The combined organic layers were washed with brine (50 mL), dried over _{anhydrous Na₂SO₄} , filtered, and concentrated under vacuum. The residue was purified by preparative TLC (DCM/MeOH = 10/1) to obtain the desired product (30 mg, yield: 16.67%). LC/MS (ESI) m/z: 369.1 [M+H] ⁺ .

¹ H NMR(400MHz,DMSO-d6)δppm 11.68(br s,1H)8.80(br s,1H)6.52(br s,1H)6.05(br s,1H)5.98(s,1H)4.33-4.34(m,1H)4.29(br d,J＝6.52Hz,1H)3.93(br dd,J＝11.16,3.01Hz,1H)3.82-3.88(m,1H)3.70-3.74(m,1H)3.61-3.65(m,1H)3.48(td,J＝11.72,2. 82Hz, 1H) 3.06 (td, J = 12.68, 3.64Hz, 1H) 2.43 (s, 3H) 2.25 (s, 3H) 2.18 (s, 3H) 1.15 (d, J = 6.64Hz, 3H).

Example 42

Biochemical tests

Test 1: ATR Suppression Test

The detection of ATR kinase activity was performed using mobility shift analysis to measure the phosphorylation of the substrate protein FAM-RAD17 (GL, catalog number: 514318, batch number: P19042-MJ524315). The analysis was developed and performed by Chempartner. All test compounds were dissolved in 20 mM 100% DMSO, and then the compounds were prepared and determined as follows:

1) Transfer 80 μl of 20 mM compound to 40 μl of 100% DMSO in a 96-well plate.

2) The compound was continuously diluted by transferring 20 μl to 60 μl of 100% DMSO into the next well, and so on, for a total of 10 concentrations.

3) Add 100 μl of 100% DMSO to two empty wells in the same 96-well plate as a compound-free control and an enzyme-free control. Label the plate as the source plate.

4) Transfer 40 μl of the compound from the source plate to a new 384-well plate as an intermediate plate.

5) Transfer 60 nmol of the compound to the assay plate using an Echo.

6) Add ATR kinase (Eurofins, catalog number: 14-953, batch number: D14JP007N) to kinase base buffer (50 mM HEPES, pH 7.5; 0.0015% Brij-35; 0.01% Triton) to prepare a 2x enzyme solution, then add 10 μl of the 2x enzyme solution to each well of a 384-well plate and incubate at room temperature for 10 minutes.

7) Add FAM-RAD17 and ATP (Sigma, catalog number: A7699-1G, CAS number: 987-65-5) to the kinase base buffer to prepare a 2x peptide solution, and then add 10 μl to the assay plate.

8) Incubate at 28°C for the specified time period. Add 40 μl of stop buffer (100 mM HEPES, pH 7.5; 0.015% Brij-35; 0.2% coating reagent #3; 50 mM EDTA) to stop the reaction.

9) Collect the data from the calipers. Convert the transformed values to suppression values.

Suppression percentage = (Maximum conversion rate) / (max - min) * 100

"max" represents the DMSO control; "min" represents the low control.

The data from XLFit Excel add-in version 5.4.0.8 was fitted to obtain the IC50 value. The formula used is:

Y = bottom + (top - bottom) / (1 + (IC50/X)^Hill slope)

Where X represents the concentration before it is converted to logarithmic form.

Table 2 below lists the IC50 values for the compounds of exemplary formula (I).

Table 2

Test 2: Tumor Cell Anti-Proliferation Test (CTG Test)

Human colorectal cancer cells HT-29 (HTB-38) and LoVo (CCL-229), both originally obtained from the American Type Culture Collection (ATCC), were selected for CTG assay. Complete culture medium was prepared by adding FBS and appropriate additives to the basal medium. The cell layer was then briefly washed with 0.25% (w/v) trypsin-0.038% (w/v) EDTA solution to remove any trace serum containing trypsin inhibitors. An appropriate volume of trypsin-EDTA solution was then added to the flask, and cells were observed under an inverted microscope until the cell layer dispersed. Finally, an appropriate volume of complete growth medium was added, and cells were gently pipetted. Cells were collected using Vi-cell XR, counted, and the cell density adjusted. Cells were then seeded into 96-well opaque-walled, clear-bottomed tissue culture plates in a CO2 incubator for 20–24 hours. All test compounds were prepared at a concentration of 10 mM in DMSO. The compound was then added to the cell culture medium in 3-fold serial dilutions, resulting in a final DMSO concentration of 0.5%. The plates were incubated at 37°C for 96 hours under 5% _CO₂ . Before measurement, an appropriate volume of CellTiter-Glo buffer was transferred to an amber vial containing CellTiter-Glo substrate to reconstitute the lyophilized enzyme/substrate mixture. Gently mixing this formed the CellTiter-Glo reagent (Promega catalog number: G7573). The plates and their contents were equilibrated to room temperature for approximately 30 minutes. Then, 100 μL of CellTiter-Glo reagent was added to the assay plate, and the contents were mixed on a vortex mixer for 2 minutes to induce cell lysis. The plates were incubated at room temperature for 10 minutes to stabilize the luminescence signal. Finally, a clear bottom was sealed with white-backed sealant, and luminescence was recorded using an Enspire. The _IC₅₀ and _GI₅₀ values were calculated using the XLFit curve fitting software with a 4-parameter logistic model: Y = bottom + (top - bottom) / (1 + (IC₅₀/X)^Hill slope).

Table 3 below provides the IC50 (Y = 50%) values for the exemplary compound (I).

Table 3

The foregoing description is intended to be merely an illustration of the basic principles of this disclosure. Furthermore, since many modifications and variations will be apparent to those skilled in the art, it is not intended to limit the invention to the exact constructions and processes described above. Therefore, all suitable modifications and equivalents can be considered to fall within the scope of the invention as defined by the appended claims.

Claims (73)

1. A compound having formula (I): Or its pharmaceutically acceptable salt. in Ring A is absent, or is a 3- to 6-membered cycloalkyl, 5- to 6-membered heterocyclic or 5- to 6-membered heteroaryl; V is a direct bond, a carbonyl group, or an alkyl group optionally substituted with one or more R ^c groups; W and L are each independently a direct bond, -O-, -S-, or -N( ^Ra )-; ^R1 is an alkyl, cyano, _{hydroxyl, -S(O)2CH3 or} -S(O)(NH) _CH3 ; ^R2 is hydrogen, halogen, or an alkyl group optionally substituted with one or more ^Rb ; Ring B is R ⁵ is selected from the group consisting of: hydrogen, halogen, hydroxyl, cyano, alkyl, alkenyl, ynyl, heteroalkyl, heteroalkenyl, heteroynyl, and haloalkyl; ^Ra is hydrogen or alkyl; ^Rb is a hydroxyl group or a halogen; ^Rc is a hydroxyl group, halogen, or alkyl group; n can be 0, 1, 2, or 3. 2. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein V is a direct bond. 3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein V is a carbonyl group. 4. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein V is an alkyl group optionally substituted with one or more R ^_c . 5. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein ring A is a 3- to 6-membered cycloalkyl group. 6. The compound of claim 5 or a pharmaceutically acceptable salt thereof, wherein ring A is cyclopropyl, cyclopentyl, or cyclohexyl, preferably cyclopropyl. 7. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein ring A is a 5- to 6-membered heterocyclic group. 8. The compound of claim 7 or a pharmaceutically acceptable salt thereof, wherein ring A is pyrazolyl or tetrahydropyranyl. 9. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein ring A is a 5- to 6-membered heteroaryl group. 10. The compound of claim 9 or a pharmaceutically acceptable salt thereof, wherein ring A is thiazolyl, triazolyl, pyridyl or isoxazolyl. 11. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein ring A is absent. 12. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein W is a direct bond. 13. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein W is -N( ^Ra )-. 14. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein ring A is absent and W is -N( ^Ra )-. 15. The compound of claim 13 or a pharmaceutically acceptable salt thereof, wherein ^Ra is hydrogen or methyl. 16. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein ring A is absent and W is a direct bond. 17. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein ^R1 is an alkyl group. 18. The compound of claim 17 or a pharmaceutically acceptable salt thereof, wherein ^R1 is a _C1-3 alkyl group. 19. The compound of claim 1 or _{a pharmaceutically acceptable salt thereof, wherein R1 is -S(O)2CH3} ^or _-S (O)(NH) _CH3 . 20. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein ^R1 is a cyano or hydroxyl group. 21. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein ring A is a 3- to 6-membered cycloalkyl, a 5- to 6-membered heterocyclic or a 5- to 6-membered heteroaryl, and R ¹ is an alkyl, hydroxyl, _-S (O) _₂CH₃ or -S(O)(NH) _CH₃ . 22. The compound of claim 21 or a pharmaceutically acceptable salt thereof, wherein ring A is cyclopropyl, cyclohexyl, tetrahydropyranyl, thiazolyl, triazolyl, pyridyl, or isoxazolyl. 23. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein ring A is a 5- or 6-membered heterocyclic group and ^R1 is an alkyl group. 24. The compound of claim 23 or a pharmaceutically acceptable salt thereof, wherein ring A is pyrazolyl, triazolyl or isoxazolyl, and ^R1 is a _C1-3 alkyl group. 25. The compound of claim 23 or a pharmaceutically acceptable salt thereof, wherein ring A is pyrazolyl, triazolyl or isoxazolyl, and ^R1 is methyl. 26. The compound of claim 1 or _a pharmaceutically acceptable salt thereof, wherein ring A is absent and ^R1 is cyano or -S(O) _2CH3 . 27. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein L is a bond. 28. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein L is -O-. 29. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein L is -S-. 30. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein L is -N( ^Ra )-. 31. The compound of claim 30 or a pharmaceutically acceptable salt thereof, wherein ^Ra is hydrogen. 32. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein ring B is... 33. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein L is -O-, -S-, or -N( ^Ra )-, and ring B is 34. The compound of claim 33 or a pharmaceutically acceptable salt thereof, wherein ^Ra is hydrogen. 35. The compound of claim 33 or a pharmaceutically acceptable salt thereof, wherein ^R5 is hydrogen or alkyl. 36. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein ^R2 is hydrogen. 37. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein ^R2 is a halogen. 38. The compound of claim 37 or a pharmaceutically acceptable salt thereof, wherein ^R2 is fluorine. 39. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein ^R2 is an alkyl group substituted with one or more ^Rb . 40. The compound of claim 39 or a pharmaceutically acceptable salt thereof, wherein ^R2 is a _C1-3 alkyl group substituted with one or more ^Rb . 41. The compound of claim 40 or a pharmaceutically acceptable salt thereof, wherein ^R2 is a methyl group substituted with one or more ^Rb . 42. The compound of claim 41 or a pharmaceutically acceptable salt thereof, wherein ^Rb is a hydroxyl group or fluorine. 43. A compound or a pharmaceutically acceptable salt thereof according to any one of the preceding claims, having formula (II) or formula (III): 44. The compound of claim 1 or a pharmaceutically acceptable salt thereof, having a formula selected from the group consisting of: as well as in U is either O or NH; V is a direct bond, a carbonyl group, or an alkyl group optionally substituted with one or more R ^c groups; W and L are each independently -O-, -S-, or -N( ^Ra )-; ^R1 is an alkyl group; ^R2 is hydrogen, halogen, or an alkyl group substituted with one or more ^Rb ; R ⁵ is hydrogen or alkyl; ^Ra is hydrogen or alkyl; ^Rb is a hydroxyl group or a halogen; and ^Rc is a hydroxyl group, a halogen, or an alkyl group. 45. The compound according to claim 1, wherein the compound is selected from the group consisting of: Or its pharmaceutically acceptable salt. 46. A compound having formula (V): Or its pharmaceutically acceptable salt. in Ring A is absent, or is a 3- to 6-membered cycloalkyl, 5- to 6-membered heterocyclic or 5- to 6-membered heteroaryl; Q is an alkyl group that is directly bonded or optionally substituted with one or more R ^d ; L is -O-, -S-, or -N( ^Ra )-; Ring B is ^Ra is hydrogen or alkyl; R ^_d is a hydroxyl group, a halogen, or an alkyl group; ^R1 is selected from the group consisting of: cyano, hydroxyl, halogen, -S( _O ) _2CH3 and -S(O)(NH) _CH3 ; R ⁵ is selected from the group consisting of: hydrogen, halogen, hydroxyl, cyano, alkyl, alkenyl, ynyl, heteroalkyl, heteroalkenyl, heteroynyl, and haloalkyl; n can be 0, 1, 2, or 3. 47. The compound of claim 46 or a pharmaceutically acceptable salt thereof, wherein Q is a direct bond. 48. The compound of claim 46 or a pharmaceutically acceptable salt thereof, wherein Q is an alkyl group. 49. The compound of claim 48 or a pharmaceutically acceptable salt thereof, wherein Q is a _C1-3 alkyl group. 50. The compound of claim 46 or a pharmaceutically acceptable salt thereof, wherein ring A is a 3- to 6-membered cycloalkyl group. 51. The compound of claim 50 or a pharmaceutically acceptable salt thereof, wherein ring A is cyclopropyl. 52. The compound of claim 51 or a pharmaceutically acceptable salt thereof, wherein ring A is... 53. The compound of claim 46 or a pharmaceutically acceptable salt thereof, wherein ring A is absent. 54. The compound of claim 46 or a pharmaceutically acceptable salt thereof, wherein ring A is a 5- or 6-membered heterocyclic group. 55. The compound of claim 46 or a pharmaceutically acceptable salt thereof, wherein ring A is a tetrahydropyranyl group. 56. The compound of claim 55 or a pharmaceutically acceptable salt thereof, wherein ring A is... 57. The compound of claim 46 or a pharmaceutically acceptable salt thereof, wherein Q is an alkyl group and ring A is absent. 58. The compound of claim 46 or a pharmaceutically acceptable salt thereof, wherein Q is a direct bond and ring A is a 3- to 6-membered cycloalkyl group or a 5- to 6-membered heterocyclic group. 59. _{The compound of claim 46 or a pharmaceutically acceptable salt thereof, wherein R1 is -S(O)2CH3} ^or _-S (O)(NH) _CH3 . 60. The compound of claim 46 or a pharmaceutically acceptable salt thereof, wherein ^R1 is a cyano, hydroxyl, or halogen. 61. The compound of claim 46 or a pharmaceutically acceptable salt thereof, wherein ring A is absent, or is a 3- to 6-membered cycloalkyl or a 5- to 6-membered heterocyclic group, and ^R1 is -S(O) _2CH3 or _-S (O)(NH) _CH3 . 62. The compound of claim 46 or a pharmaceutically acceptable salt thereof, wherein ring A is absent or is a 3- to 6-membered cycloalkyl group, and ^R1 is a cyano, hydroxyl, or halogen. 63. The compound of claim 46 or a pharmaceutically acceptable salt thereof, wherein L is -O-. 64. The compound of claim 46 or a pharmaceutically acceptable salt thereof, wherein L is -S-. 65. The compound of claim 46 or a pharmaceutically acceptable salt thereof, wherein L is -N( ^Ra )- and ^Ra is hydrogen. 66. The compound according to any one of claims 63 to 65, or a pharmaceutically acceptable salt thereof, wherein ring B is... 67. The compound of claim 46 or a pharmaceutically acceptable salt thereof, wherein ^R5 is hydrogen or alkyl. 68. The compound according to claim 46, wherein the compound is selected from the group consisting of: Or its pharmaceutically acceptable salt. 69. A pharmaceutical composition comprising a compound according to any one of claims 1 to 68 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier. 70. A method for treating cancer, the method comprising administering to a subject in need an effective amount of a compound according to any one of claims 1 to 68 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition according to claim 69. 71. Use of a compound according to any one of claims 1 to 68, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 69, in the preparation of a medicament for treating cancer. 72. The compound of any one of claims 1 to 68 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 69, for the treatment of cancer. 73. A method for inhibiting ATR kinase in a subject in need, the method comprising administering to the subject an effective amount of a compound according to any one of claims 1 to 68 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition according to claim 69.