WO2023138621A1

WO2023138621A1 - Atr inhibitors and uses thereof

Info

Publication number: WO2023138621A1
Application number: PCT/CN2023/072923
Authority: WO
Inventors: Bo Shan; Bing HOU; Hui YUWEN; Peng Chen; Zhongyang SHI; Feifei YANG; Xingxing Wang; Jay Mei
Original assignee: Shanghai Antengene Corporation Limited; Antengene Discovery Limited
Priority date: 2022-01-19
Filing date: 2023-01-18
Publication date: 2023-07-27
Also published as: TW202344251A

Abstract

The present disclosure relates to novel compounds useful as inhibitors of ATR kinase, as well as pharmaceutical compositions comprising these compounds and methods of treatment by administration of these compounds or the pharmaceutical compositions.

Description

ATR INHIBITORS AND USES THEREOF

FIELD OF THE DISCLOSURE

The present disclosure generally relates to novel compounds useful as ATR inhibitors, as well as pharmaceutical compositions comprising these compounds and methods of treatment by administration of these compounds or the pharmaceutical compositions.

BACKGROUND OF THE DISCLOSURE

ATR (also known as FRAP-Related Protein 1; FRP1, MEC1, SCKL, SECKL1) protein kinase is a member of the PI3-Kinase like kinase (PIKK) family of proteins involved in repair and maintenance of the genome and its stability. It is essential to the viability of replicating cells and is activated during S-phase to regulate firing of replication origins and to repair damaged replication forks. Therefore, ATR inhibitors have the potential to be an efficient way in cancer treatment.

While progress has been made for ATR inhibitors, there is still a strong need in the art to develop improved pharmaceutics having inhibitory activity against ATR.

SUMMARY OF THE DISCLOSURE

The present disclosure provides compounds, including stereoisomers, pharmaceutically acceptable salts, tautomers and prodrugs thereof, which are capable of inhibiting ATR protein kinase. Methods for use of such compounds for treatment of various diseases or conditions, such as cancer, are also provided.

In one aspect, the present disclosure provides a compound having Formula (I) :

or a pharmaceutically acceptable salt thereof,

wherein

R¹ and R² each independently is alkyl, alkenyl, alkynyl, heteroaryl, heteroalkenyl, heteroalkynyl, cycloalkyl, heterocyclyl, aryl or heteroaryl, wherein said cycloalkyl, heterocyclyl, aryl and heteroaryl are optionally substituted with one or more R^a; or

R¹ and R² together with the sulfur atom to which they are both attached form a heterocyclyl optionally substituted with one or more R^b;

R^a is independently selected from the group consisting of hydroxyl, halogen, cyano, amino, alkyl, alkoxyl, and haloalkyl;

R^b is independently selected from the group consisting of alkyl, alkenyl, alknyl, alkoxy, cyano, halogen, hydroxyl, amino, and nitro, wherein said alkyl, alkenyl and alknyl are optionally substituted with one or more groups independently selected from hydroxyl, halogen, cyano, amino and alkoxyl;

R³ is

Ring A is 5-to 6-membered heterocyclyl or 5-to 6-membered heteroaryl;

R⁴, in each occurrence, is halogen, alkyl, haloalkyl, or cycloalkyl; and

m is 0, 1, 2 or 3.

In some embodiments, the present disclosure provides compound having a formula of (Ia) :

or a pharmaceutically acceptable salt thereof.

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

or a pharmaceutically acceptable salt thereof.

In another aspect, the present disclosure provides a compound selected from the group consisting of:

or a pharmaceutically acceptable salt thereof.

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

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

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

In a further aspect, the present disclosure provides compounds of the present disclosure or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of the present disclosure, for use in the treatment of cancer.

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

DETAILED DESCRIPTION OF THE DISCLOSURE

Reference will now be made in detail to certain embodiments of the present disclosure, examples of which are illustrated in the accompanying structures and formulas. While the present disclosure will be described in conjunction with the enumerated embodiments, it will be understood that they are not intended to limit the present disclosure to those embodiments. On the contrary, the present disclosure is intended to cover all alternatives, modifications, and equivalents, which may be included within the scope of the present disclosure as defined by the claims. One skilled in the art will recognize many methods and materials similar or equivalent to those described herein, which could be used in the practice of the present disclosure. The present disclosure is in no way limited to the methods and materials described. In the event that one or more of the incorporated references and similar materials differs from or contradicts this application, including but not limited to defined terms, term usage, described techniques, or the like, the present disclosure controls. All references, patents, patent applications cited in the present disclosure are hereby incorporated by reference in their entireties.

It is appreciated that certain features of the present disclosure, which are, for clarity, described in the context of separate embodiments, can also be provided in combination in a single embodiment. Conversely, various features of the present disclosure, which are, for brevity, described in the context of a single embodiment, can also be provided separately or in any suitable sub-combination. It must be noted that, as used in the specification and the appended claims, the singular forms “a, ” “an, ” and “the” include plural forms of the same unless the context clearly dictates otherwise. Thus, for example, reference to “a compound” includes a plurality of compounds.

Definitions

Definitions of specific functional groups and chemical terms are described in more detail below. For purposes of this disclosure, the chemical elements are identified in accordance with the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75^th Ed., inside cover, and specific functional groups are generally defined as described therein. Additionally, general principles of organic chemistry, as well as specific functional moieties and reactivity, are described in Organic Chemistry, Thomas Sorrell, 2^nd Edition, University Science Books, Sausalito, 2006; Smith and March March’s Advanced Organic Chemistry, 6^th Edition, John Wiley &Sons, Inc., New York, 2007; Larock, Comprehensive Organic Transformations, 3^rd Edition, VCH Publishers, Inc., New York, 2018; Carruthers, Some Modern Methods of Organic Synthesis, 4^th Edition, Cambridge University Press, Cambridge, 2004; the entire contents of each of which are incorporated herein by reference.

Generally, the nomenclature used herein and the laboratory procedures in organic chemistry, medicinal chemistry, and pharmacology described herein are those well known and commonly employed in the art. Unless defined otherwise, all technical and scientific terms used herein generally have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs.

It should be noted that if there is a discrepancy between a depicted structure and a name given that structure, the depicted structure is to be accorded more weight. In addition, if the stereochemistry of a structure or a portion of a structure is not indicated with, for example, bold or dashed lines, the structure or portion of the structure is to be interpreted as encompassing all stereoisomers of it.

At various places in the present disclosure, linking substituents are described. Where the structure clearly requires a linking group, the Markush variables listed for that group are understood to be linking groups. For example, if the structure requires a linking group and the Markush group definition for that variable lists “alkyl” , then it is understood that the “alkyl” represents a linking alkylene group.

When a bond to a substituent is shown to cross a bond connecting two atoms in a ring, then such substituent may be bonded to any atom in the ring. When a substituent is listed without indicating the atom via which such substituent is bonded to the rest of the compound of a given formula, then such substituent may be bonded via any atom in such formula. Combinations of substituents and/or variables are permissible, but only if such combinations result in stable compounds.

When any variable (e.g., Rⁱ) occurs more than one time in any constituent or formula for a compound, its definition at each occurrence is independent of its definition at every other occurrence. Thus, for example, if a group is shown to be substituted with 0-2 Rⁱ moieties, then the group may optionally be substituted with up to two Rⁱ moieties and Rⁱ at each occurrence is selected independently from the definition of Rⁱ. Also, combinations of substituents and/or variables are permissible, but only if such combinations result in stable compounds.

As used herein, the term “C_i-j” indicates a range of the carbon atoms numbers, wherein i and j are integers and the range of the carbon atoms numbers includes the endpoints (i.e. i and j) and each integer point in between, and wherein j is greater than i. For examples, 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, particularly 1 to 10, particularly 1 to 8, particularly 1 to 6, particularly 1 to 5, particularly 1 to 4, particularly 1 to 3 or particularly 1 to 2 carbon atoms.

As used herein, the term “alkyl” , whether as part of another term or used independently, refers to a saturated linear or branched-chain hydrocarbon radical, which may be optionally substituted independently with one or more substituents described below. The term “C_i-j alkyl” refers to an alkyl having i to j carbon atoms. In some embodiments, alkyl groups contain 1 to 10 carbon atoms. In some embodiments, alkyl groups contain 1 to 9 carbon atoms. In some embodiments, alkyl groups contain 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” are methyl, ethyl, propyl, isopropyl, n-butyl, i-butyl, s-butyl, t-butyl, n-pentyl, 2-pentyl, 3-pentyl, 2-methyl-2-butyl, 3-methyl-2-butyl, 3-methyl-1-butyl, 2-methyl-1-butyl, 1-hexyl, 2- hexyl, 3-hexyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 3-methyl-3-pentyl, 2-methyl-3-pentyl, 2, 3-dimethyl-2-butyl, 3, 3-dimethyl-2-butyl, and the like.

As used herein, the term “alkoxyl” , whether as part of another term or used independently, refers to an alkyl group, as previously defined, attached to the parent molecule through an oxygen atom. The term “C_i-j alkoxy” means that the alkyl moiety of the alkoxy group has i to j carbon atoms. In some embodiments, alkoxy groups contain 1 to 10 carbon atoms. In some embodiments, alkoxy groups contain 1 to 9 carbon atoms. In some embodiments, alkoxy groups contain 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_1-6 alkoxyl” include, but are not limited to, methoxy, ethoxy, propoxy (e.g. n-propoxy and isopropoxy) , t-butoxy, neopentoxy, n-hexoxy, and the like.

As used herein, the term “amino” refers to –NH₂. Amino groups may also be substituted with one or more groups such as alkyl, aryl, carbonyl or other amino groups.

As used herein, the term “aryl” , whether as part of another term or used independently, refers to monocyclic and polycyclic ring systems having a total of 5 to 20 ring members, wherein at least one ring in the system is aromatic and wherein each ring in the system contains 3 to 12 ring members. Examples of “aryl” include, but are not limited to, phenyl, biphenyl, naphthyl, anthracyl and the like, which may bear one or more substituents. Also included within the scope of the term “aryl” , as it is used herein, is a group in which an aromatic ring is fused to one or more additional rings. In the case of polycyclic ring system, only one of the rings needs to be aromatic (e.g., 2, 3-dihydroindole) , although all of the rings may be aromatic (e.g., quinoline) . The second ring can also be fused or bridged. Examples of polycyclic aryl include, but are not limited to, benzofuranyl, indanyl, phthalimidyl, naphthimidyl, phenanthridinyl, or tetrahydronaphthyl, and the like. Aryl groups can be substituted at one or more ring positions with substituents as described above.

As used herein, the term “cycloalkyl” , whether as part of another term or used independently, refer to a monovalent non-aromatic, saturated or partially unsaturated monocyclic and polycyclic ring system, in which all the ring atoms are carbon and which contains at least three ring forming carbon atoms. In some embodiments, the cycloalkyl may contain 3 to 12 ring forming carbon atoms, 3 to 10 ring forming carbon atoms, 3 to 9 ring forming carbon atoms, 3 to 8 ring forming carbon atoms, 3 to 7 ring forming carbon atoms, 3 to 6 ring forming carbon atoms, 3 to 5 ring forming carbon atoms, 4 to 12 ring forming carbon atoms, 4 to 10 ring forming carbon atoms, 4 to 9 ring forming carbon atoms, 4 to 8 ring forming carbon atoms, 4 to 7 ring forming carbon atoms, 4 to 6 ring forming carbon atoms, 4 to 5 ring forming carbon atoms. Cycloalkyl groups may be saturated or partially unsaturated. Cycloalkyl groups may be substituted. In some embodiments, the cycloalkyl group may be a saturated cyclic alkyl group. In some embodiments, the cycloalkyl group may be a partially unsaturated cyclic alkyl group that contains at least one double bond or triple bond in its ring system. In some embodiments, the cycloalkyl group may be monocyclic or polycyclic. Examples of monocyclic cycloalkyl group 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 group include, but are not limited to, adamantyl, norbornyl, fluorenyl, spiro-pentadienyl, spiro [3.6] -decanyl, bicyclo [1, 1, 1] pentenyl, bicyclo [2, 2, 1] heptenyl, and the like.

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, as defined above, that is substituted by one or more halogens, as defined above. Examples of haloalkyl include, but are not limited to, trifluoromethyl, difluoromethyl, trichloromethyl, 2, 2, 2-trifluoroethyl, 1, 2-difluoroethyl, 3-bromo-2-fluoropropyl, 1, 2-dibromoethyl, and the like.

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 a basic nitrogen (including N-oxides) .

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

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

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

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

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

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

As used herein, the term “hydroxyl” refers to –OH.

As used herein, the term “partially unsaturated” refers to a radical that includes at least one double or triple bond. The term “partially unsaturated” is intended to encompass rings having multiple sites of unsaturation, but is not intended to include aromatic (i.e., fully unsaturated) moieties.

As used herein, the term “substituted” , whether preceded by the term “optionally” or not, means that one or more hydrogens of the designated moiety are replaced with a suitable substituent. It will be understood that “substitution” or “substituted with” includes the implicit proviso that such substitution is in accordance with permitted valence of the substituted atom and that the substitution results in a stable or chemically feasible compound, e.g., which does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, etc. Unless otherwise indicated, an “optionally substituted” group may have a suitable substituent at each substitutable position of the group, and when more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituent may be either the same or different at every position. It will be understood by those skilled in the art that substituents can themselves be substituted, if appropriate. Unless specifically stated as “unsubstituted” , references to chemical moieties herein are understood to include substituted variants. For example, reference to an “aryl” group or moiety implicitly includes both substituted and unsubstituted variants.

Compounds

The present disclosure provides novel compounds of Formula (I) and pharmaceutically acceptable salts thereof, synthetic methods for making the compounds, pharmaceutical compositions containing them and various uses of the disclosed compounds.

In one aspect, the present disclosure provides a compound having Formula (I) :

or a pharmaceutically acceptable salt thereof,

wherein

R³ is

Ring A is 5-to 6-membered heterocyclyl or 5-to 6-membered heteroaryl;

R⁴, in each occurrence, is halogen, alkyl, haloalkyl, or cycloalkyl; and

m is 0, 1, 2 or 3.

In some embodiments, R¹ and R² are each independently C_1-3 alkyl, C_3-6 cycloalkyl or C_5-12 aryl.

In some embodiments, R¹ and R² are independently C_1-3 alkyl. In certain embodiments, R¹ and R² are the same. In certain embodiments, both R¹ and R² are methyl. In certain embodiments, R¹ and R² are different. In certain embodiments, one of R¹ and R² is methyl, and the other is ethyl.

In some embodiments, R¹ and R² are independently C_3-6 cycloalkyl. In certain embodiments, both R¹ and R² are cyclopropyl.

In some embodiments, one of R¹ and R² is C_1-3 alkyl, and the other is C_3-6 cycloalkyl or C_5-12 aryl.

In some embodiments, is selected from the group consisting of:

In some embodiments, R¹ and R² together with the sulfur atom to which they are both attached form a heterocyclyl optionally substituted with one or more R^b.

In some embodiments, R¹ and R² together with the sulfur atom to which they are both attached form a 3-to 6-membered heterocyclyl optionally substituted with one or more R^b.

In some embodiments, is selected from each of which is optionally substituted with one or more R^b.

In some embodiments, Ring A is 5-to 6-membered heteroaryl. In certain embodiments, Ring A is pyrazolyl, pyrrolyl, or pyridyl.

In some embodiments, R⁴ is halogen.

In some embodiments, R⁴ is alkyl. In certain embodiments, R⁴ is C_1-3 alkyl.

In some embodiments, R⁴ is haloalkyl. In certain embodiments, R⁴ is C_1-3 haloalkyl.

In some embodiments, R⁴ is cycloalkyl. In certain embodiments, R⁴ is C_3-6 cycloalkyl.

In some embodiments, m is 0, 1 or 2.

In some embodiments, is selected from the group consisting of:

In some embodiments, R³ is

In some embodiments, the present disclosure provides a compound having a formula (Ia) :

or a pharmaceutically acceptable salt thereof.

In some embodiments, the present disclosure provides a compound selected from the group consisting of:

or a pharmaceutically acceptable salt thereof.

In a further aspect, the present disclosure provides a compound selected from the group consisting of:

or a pharmaceutically acceptable salt thereof.

Exemplary compounds of the present disclosure are set forth in Table 1 below.

Table 1

Compounds provided herein are described with reference to both generic formulae and specific compounds. In addition, the compounds of the present disclosure may exist in a number of different forms or derivatives, including but not limited to prodrugs, soft drugs, active metabolic derivatives (active metabolites) , and their pharmaceutically acceptable salts, all within the scope of the present disclosure.

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

As used herein, the term “soft drug” refers to compounds that exert a pharmacological effect but break down to inactive metabolites degradants so that the activity is of limited time. See, for example, “Soft drugs: Principles and methods for the design of safe drugs” , Nicholas Bodor, Medicinal Research Reviews, Vol. 4, No. 4, 449-469, 1984, which is hereby incorporated by reference in its entirety.

As used herein, the term “metabolite” , e.g., active metabolite overlaps with prodrug as described above. Thus, such metabolites are pharmacologically active compounds or compounds that further metabolize to pharmacologically active compounds that are derivatives resulting from metabolic process in the body of a subject. For example, such metabolites may result from oxidation, reduction, hydrolysis, amidation, deamidation, esterification, deesterification, enzymatic cleavage, and the like, of the administered compound or salt or prodrug. Of these, active metabolites are such pharmacologically active derivative compounds. For prodrugs, the prodrug compound is generally inactive or of lower activity than the metabolic product. For active metabolites, the parent compound may be either an active compound or may be an inactive prodrug.

Prodrugs and active metabolites may be identified using routine techniques know in the art. See, e.g., Bertolini et al, 1997, J Med Chem 40: 2011-2016; Shan et al., J Pharm Sci 86: 756-757; Bagshawe, 1995, DrugDev Res 34: 220-230; Wermuth, supra.

As used herein, the term “pharmaceutically acceptable” indicates that the substance or composition is compatible chemically and/or toxicologically, with the other ingredients comprising a formulation, and/or the subjects being treated therewith.

As used herein, the term “pharmaceutically acceptable salt” , unless otherwise indicated, includes salts that retain the biological effectiveness of the free acids and bases of the specified compound and that are not biologically or otherwise undesirable. Contemplated pharmaceutically acceptable salt forms include, but are not limited to, mono, bis, tris, tetrakis, and so on. Pharmaceutically acceptable salts are non-toxic in the amounts and concentrations at which they are administered. The preparation of such salts can facilitate the pharmacological use by altering the physical characteristics of a compound without preventing it from exerting its physiological effect. Useful alterations in physical properties include lowering the melting point to facilitate transmucosal administration and increasing the solubility to facilitate administering higher concentrations of the drug.

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

Pharmaceutically acceptable salts also include basic addition salts such as those containing benzathine, chloroprocaine, choline, diethanolamine, ethanolamine, t-butylamine, ethylenediamine, meglumine, procaine, aluminum, calcium, lithium, magnesium, potassium, sodium, ammonium, alkylamine, and zinc, when acidic functional groups, such as carboxylic acid or phenol are present. For example, see Remington's Pharmaceutical Sciences, 19^thed., Mack Publishing Co., Easton, PA, Vol. 2, p. 1457, 1995; “Handbook of Pharmaceutical Salts: Properties, Selection, and Use” by Stahl and Wermuth, Wiley-VCH, Weinheim, Germany, 2002. Such salts can be prepared using the appropriate corresponding bases.

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

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

It is also to be understood that the compounds of present disclosure can exist in unsolvated forms, solvated forms (e.g., hydrated forms) , and solid forms (e.g., crystal or polymorphic forms) , and the present disclosure is intended to encompass all such forms.

As used herein, the term “solvate” or “solvated form” refers to solvent addition forms that contain either stoichiometric or non-stoichiometric amounts of solvent. Some compounds have a tendency to trap a fixed molar ratio of solvent molecules in the crystalline solid state, thus forming a solvate. If the solvent is water the solvate formed is a hydrate; and if the solvent is alcohol, the solvate formed is an alcoholate. Hydrates are formed by the combination of one or more molecules of water with one molecule of the substance in which the water retains its molecular state as H₂O. 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 “crystal form” , “crystalline form” , “polymorphic forms” and “polymorphs” can be used interchangeably, and mean crystal structures in which a compound (or a salt or solvate thereof) can crystallize in different crystal packing arrangements, all of which have the same elemental composition. Different crystal forms usually have different X-ray diffraction patterns, infrared spectral, melting points, density hardness, crystal shape, optical and electrical properties, stability and solubility. Recrystallization solvent, rate of crystallization, storage temperature, and other factors may cause one crystal form to dominate. Crystal polymorphs of the compounds can be prepared by crystallization under different conditions.

The compounds of present disclosure can comprise one or more asymmetric centers depending on substituent selection, and thus can exist in various stereoisomeric forms, e.g., enantiomers and/or diastereomers. For example, the compounds provided herein may have an asymmetric carbon center, and thus compounds provided herein may have either the (R) or (S) stereo-configuration at a carbon asymmetric center. Therefore, compounds of the present disclosure may be in the form of an individual enantiomer, diastereomer or geometric isomer, or may be in the form of a mixture of stereoisomers.

As used herein, the term “enantiomer” refers to two stereoisomers of a compound which are non-superimposable mirror images of one another. The term “diastereomer” refers to a pair of optical isomers which are not mirror images of one another. Diastereomers have different physical properties, e.g. melting points, boiling points, spectral properties, and reactivities.

Where a particular enantiomer is preferred, it may, in some embodiments be provided substantially free of the opposite enantiomer, and may also be referred to as “optically enriched” . “Optically enriched” , as used herein, means that the compound is made up of a significantly greater proportion of one enantiomer. In certain embodiments, the compound is made up of at least about 90%by weight of a preferred enantiomer. In other embodiments, the compound is made up of at least about 95%, 98%, or 99%by weight of a preferred enantiomer. Preferred enantiomers may be isolated from racemic mixtures by any method known to those skilled in the art, for example by chromatography or crystallization, by the use of stereochemically uniform starting materials for the synthesis or by stereoselective synthesis. Optionally a derivatization can be carried out before a separation of stereoisomers. The separation of a mixture of stereoisomers can be carried out at an intermediate step during the synthesis of a compound provided herein or it can be done on a final racemic product. Absolute stereochemistry may be determined by X-ray crystallography of crystalline products or crystalline intermediates which are derivatized, if necessary, with a reagent containing a stereogenic center of known configuration. Alternatively, absolute stereochemistry may be determined by Vibrational Circular Dichroism (VCD) spectroscopy analysis. 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, Notre Dame, IN 1972) .

In some embodiments, mixtures of diastereomers, for example mixtures of diastereomers enriched with 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 are provided.

In some embodiments, compounds provided herein may have one or more double bonds that can exist as either the Z or E isomer, unless otherwise indicated. The present disclosure additionally encompasses the compounds as individual isomers substantially free of other isomers and alternatively, as mixtures of various isomers, e.g., racemic mixtures of enantiomers.

The compounds of the present disclosure may also exist in different tautomeric forms, and all such forms are embraced within the scope of the present disclosure. The term “tautomer” or “tautomeric form” refers to structural isomers of different energies which are interconvertible via a low energy barrier. For example, proton tautomers (also known as prototropic tautomers) include interconversions via migration of a proton, such as keto-enol, amide-imidic acid, lactam-lactim, imine-enamine isomerizations and annular forms where a proton can occupy two or more positions of a heterocyclic system (for example, 1H-and 3H-imidazole, 1H-, 2H-and 4H-1, 2, 4-triazole, 1H-and 2H-isoindole, and 1H-and 2H-pyrazole) . Valence tautomers include interconversions by reorganization of some of the bonding electrons. Tautomers can be in equilibrium or sterically locked into one form by appropriate substitution. Compounds of the present disclosure identified by name or structure as one particular tautomeric form are intended to include other tautomeric forms unless otherwise specified.

The present disclosure is also intended to include all isotopes of atoms in the compounds. Isotopes of an atom include atoms having the same atomic number but different mass numbers. For example, unless otherwise specified, hydrogen, carbon, nitrogen, oxygen, phosphorous, sulfur, fluorine, chlorine, bromide or iodine in the compounds of present disclosure are meant 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 embodiments, carbon includes ¹²C and ¹³C.

Synthesis of compounds

Synthesis of the compounds provided herein, including pharmaceutically acceptable salts thereof, are illustrated in the synthetic schemes in the examples. The compounds provided herein can be prepared using any known organic synthesis techniques and can be synthesized according to any of numerous possible synthetic routes, and thus these schemes are illustrative only and are not meant to limit other possible methods that can be used to prepare the compounds provided herein. Additionally, the steps in the Schemes are for better illustration and can be changed as appropriate. The embodiments of the compounds in examples were synthesized for the purposes of research and potentially submission to regulatory agencies.

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

Preparation of compounds of the present disclosure can 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 one skilled in the art. The chemistry of protecting groups can be found, for example, in T.W. Greene and P.G.M. Wuts, Protective Groups in Organic Synthesis, 3rd Ed., Wiley &Sons, Inc., New York (1999) , in P. Kocienski, Protecting Groups, Georg Thieme Verlag, 2003, and in Peter G.M. Wuts, Greene's Protective Groups in Organic Synthesis, 5^th Edition, Wiley, 2014, all of which are incorporated herein by reference in its entirety.

Reactions can be monitored according to 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-visible) , mass spectrometry, or by chromatographic methods such as high performance liquid chromatography (HPLC) , liquid chromatography-mass spectroscopy (LCMS) , or thin layer chromatography (TLC) . Compounds can be purified by one skilled in the art by a variety of methods, including high performance liquid chromatography (HPLC) ( “Preparative LC-MS Purification: Improved Compound Specific Method Optimization” Karl F. Blom, Brian Glass, Richard Sparks, Andrew P. Combs J. Combi. Chem. 2004, 6 (6) , 874-883, which is incorporated herein by reference in its entirety) , and normal phase silica chromatography.

The known starting materials of the present disclosure can be synthesized by using or according to the known methods in the art, or can be purchased from commercial suppliers. Unless otherwise noted, analytical grade solvents and commercially available reagents were used without further purification.

Unless otherwise specified, the reactions of the present disclosure were all done under a positive pressure of nitrogen or argon or with a drying tube in anhydrous solvents, and the reaction flasks were typically fitted with rubber septa for the introduction of substrates and reagents via syringe. Glassware was oven dried and/or heat dried.

For illustrative purposes, the Examples section below shows synthetic route for preparing the compounds of the present disclosure as well as key intermediates. Those skilled in the art will appreciate that other synthetic routes may be used to synthesize the inventive compounds. Although specific starting materials and reagents are depicted, other starting materials and reagents can be easily substituted to provide a variety of derivatives and/or reaction conditions. In addition, many of the compounds prepared by the methods described below can be further modified in light of this disclosure using conventional chemistry well known to those skilled in the art.

Pharmaceutical Compositions

In a further aspect, there is provided pharmaceutical compositions comprising one or more molecules or compounds of the present disclosure, or a pharmaceutically acceptable salt thereof.

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

As used herein, the term “pharmaceutical composition” refers to a formulation containing the molecules or compounds of the present disclosure in a form suitable for administration to a subject.

As used herein, the term “pharmaceutically acceptable excipient” means an excipient that is useful in preparing a pharmaceutical composition that is generally safe, non-toxic and neither biologically nor otherwise undesirable, and includes excipient that is acceptable for veterinary use as well as human pharmaceutical use. A “pharmaceutically acceptable excipient” as used herein includes both one and more than one such excipient. The term “pharmaceutically acceptable excipient” also encompasses “pharmaceutically acceptable carrier” and “pharmaceutically acceptable diluent” .

The particular excipient used will depend upon the means and purpose for which the compounds of the present disclosure is being applied. Solvents are generally selected based on solvents recognized by persons skilled in the art as safe to be administered to a mammal including humans. In general, safe solvents are non-toxic aqueous solvents such as water and other non-toxic solvents that are soluble or miscible in water. Suitable aqueous solvents include water, ethanol, propylene glycol, polyethylene glycols (e.g., PEG 400, PEG 300) , etc. and mixtures thereof.

In some embodiments, suitable excipients may include buffers such as phosphate, citrate and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride, benzethonium chloride; phenol, butyl or benzyl alcohol; alkyl parabens such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol) ; low molecular weight (less than about 10 residues) polypeptides; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose or sorbitol; salt-forming counter-ions such as sodium; metal complexes (e.g., Zn-protein complexes) ; and/or non-ionic surfactants such as TWEEN^TM, PLURONICS^TM or polyethylene glycol (PEG) .

In some embodiments, suitable excipients may include one or more stabilizing agents, surfactants, wetting agents, lubricating agents, emulsifiers, suspending agents, preservatives, antioxidants, opaquing agents, glidants, processing aids, colorants, sweeteners, perfuming agents, flavoring agents and other known additives to provide an elegant presentation of the drug (i.e., a compound of the present disclosure or pharmaceutical composition thereof) or aid in the manufacturing of the pharmaceutical product (i.e., medicament) . The active pharmaceutical ingredients may also be entrapped in microcapsules prepared, for example, by coacervation techniques or by interfacial polymerization, for example, hydroxymethylcellulose or gelatin-microcapsules and poly- (methylmethacylate) microcapsules, respectively, in colloidal drug delivery systems (for example, liposomes, albumin microspheres, microemulsions, nano-particles and nanocapsules) or in macroemulsions. Such techniques are disclosed in Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980) . A “liposome” is a small vesicle composed of various types of lipids, phospholipids and/or surfactant which is useful for delivery of a drug (such as the compounds disclosed herein and, optionally, a chemotherapeutic agent) to a mammal including humans. The components of the liposome are commonly arranged in a bilayer formation, similar to the lipid arrangement of biological membranes.

The pharmaceutical compositions provided herein can be in any form that allows for the composition to be administered to a subject, including, but not limited to a human, and formulated to be compatible with an intended route of administration.

A variety of routes are contemplated for the pharmaceutical compositions provided herein, and accordingly the pharmaceutical composition provided herein may be supplied in bulk or in unit dosage form depending on the intended administration route. For example, for oral, buccal, and sublingual administration, powders, suspensions, granules, tablets, pills, capsules, gelcaps, and caplets may be acceptable as solid dosage forms, and emulsions, syrups, elixirs, suspensions, and solutions may be acceptable as liquid dosage forms. For injection administration, emulsions and suspensions may be acceptable as liquid dosage forms, and a powder suitable for reconstitution with an appropriate solution as solid dosage forms. For inhalation administration, solutions, sprays, dry powders, and aerosols may be acceptable dosage form. For topical (including buccal and sublingual) or transdermal administration, powders, sprays, ointments, pastes, creams, lotions, gels, solutions, and patches may be acceptable dosage form. For vaginal administration, pessaries, tampons, creams, gels, pastes, foams and spray may be acceptable dosage form.

The quantity of active ingredient in a unit dosage form of composition is a therapeutically effective amount and is varied according to the particular treatment involved. As used herein, the term “therapeutically effective amount” refers to an amount of a molecule, compound, or composition comprising the molecule or compound to treat, ameliorate, or prevent an identified disease or condition, or to exhibit a detectable therapeutic or inhibitory effect. The effect can be detected by any assay method known in the art. The precise effective amount for a subject will depend upon the subject’s body weight, size, and health; the nature and extent of the condition; the rate of administration; the therapeutic or combination of therapeutics selected for administration; and the discretion of the prescribing physician. Therapeutically effective amounts for a given situation can be determined by routine experimentation that is within the skill and judgment of the clinician.

In some embodiments, the pharmaceutical compositions of the present disclosure may be in a form of formulation for oral administration.

In certain embodiments, the pharmaceutical compositions of the present disclosure may be in the form of tablet formulations. Suitable pharmaceutically- acceptable excipients for a tablet formulation include, for example, inert diluents such as lactose, sodium carbonate, calcium phosphate or calcium carbonate, granulating and disintegrating agents such as corn starch or algenic acid; binding agents such as starch; lubricating agents such as magnesium stearate, stearic acid or talc; preservative agents such as ethyl or propyl p-hydroxybenzoate, and anti-oxidants, such as ascorbic acid. Tablet formulations may be uncoated or coated either to modify their disintegration and the subsequent absorption of the active ingredient within the gastrointestinal tract, or to improve their stability and/or appearance, in either case using conventional coating agents and procedures well known in the art.

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

In certain embodiments, the pharmaceutical compositions of the present disclosure may be in the form of aqueous suspensions, which generally contain the active ingredient in finely powdered form together with one or more suspending agents, such as sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinyl-pyrrolidone, gum tragacanth and gum acacia; dispersing or wetting agents such as lecithin or condensation products of an alkylene oxide with fatty acids (for example polyoxethylene stearate) , or condensation products of ethylene oxide with long chain aliphatic alcohols, for example heptadecaethyleneoxycetanol, or condensation products of ethylene oxide with partial esters derived from fatty acids and a hexitol such as polyoxyethylene sorbitol monooleate, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, for example polyethylene sorbitan monooleate. The aqueous suspensions may also contain one or more preservatives (such as ethyl or propyl p-hydroxybenzoate, anti-oxidants (such as ascorbic acid) , coloring agents, flavoring agents, and/or sweetening agents (such as sucrose, saccharine or aspartame) .

In certain embodiments, the pharmaceutical compositions of the present disclosure may be in the form of oily suspensions, which generally contain suspended active ingredient in a vegetable oil (such as arachis oil, olive oil, sesame oil or coconut oil) or in a mineral oil (such as liquid paraffin) . The oily suspensions may also contain a thickening agent such as beeswax, hard paraffin or cetyl alcohol. Sweetening agents such as those set out above, and flavoring agents may be added to provide a palatable oral preparation. These compositions may be preserved by the addition of an anti-oxidant such as ascorbic acid.

In certain embodiments, the pharmaceutical compositions of the present disclosure may be in the form of oil-in-water emulsions. The oily phase may be a vegetable oil, such as olive oil or arachis oil, or a mineral oil, such as for example liquid paraffin or a mixture of any of these. Suitable emulsifying agents may be, for example, naturally-occurring gums such as gum acacia or gum tragacanth, naturally-occurring phosphatides such as soya bean, lecithin, esters or partial esters derived from fatty acids and hexitol anhydrides (for example sorbitan monooleate) and condensation products of the said partial esters with ethylene oxide such as polyoxyethylene sorbitan monooleate. The emulsions may also contain sweetening, flavoring and preservative agents.

In certain embodiments, the pharmaceutical compositions provided herein may be in the form of syrups and elixirs, which may contain sweetening agents such as glycerol, propylene glycol, sorbitol, aspartame or sucrose, a demulcent, a preservative, a flavoring and/or coloring agent.

In some embodiments, the pharmaceutical compositions of the present disclosure may be in a form of formulation for injection administration.

In certain embodiments, the pharmaceutical compositions of the present disclosure may be in the form of a sterile injectable preparation, such as a sterile injectable aqueous or oleaginous suspension. This suspension may be formulated according to the known art using those suitable dispersing or wetting agents and suspending agents, which have been mentioned above. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, such as a solution in 1, 3-butanediol or prepared as a lyophilized powder. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile fixed oils may conventionally be employed as a solvent or suspending medium. For this purpose any bland fixed oil may be employed including synthetic mono-or diglycerides. In addition, fatty acids such as oleic acid may likewise be used in the preparation of injectables.

In some embodiments, the pharmaceutical compositions of the present disclosure may be in a form of formulation for inhalation administration.

In certain embodiments, the pharmaceutical compositions of the present disclosure may be in the form of aqueous and nonaqueous (e.g., in a fluorocarbon propellant) aerosols containing any appropriate solvents and optionally other compounds such as, but not limited to, stabilizers, antimicrobial agents, antioxidants, pH modifiers, surfactants, bioavailability modifiers and combinations of these. The carriers and stabilizers vary with the requirements of the particular compound, but typically include nonionic surfactants (Tweens, Pluronics, or polyethylene glycol) , innocuous proteins like 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 the present disclosure may be in a form of formulation for topical or transdermal administration.

In certain embodiments, the pharmaceutical compositions provided herein may be in the form of creams, ointments, gels and aqueous or oily solutions or suspensions, which may generally be obtained by formulating an active ingredient with a conventional, topically acceptable excipients such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.

In certain embodiments, the pharmaceutical compositions provided herein may be formulated in the form of transdermal skin patches that are well known to those of ordinary skill in the art.

Besides those representative dosage forms described above, pharmaceutically acceptable excipients and carriers are generally known to those skilled in the art and are thus included in the present disclosure. Such excipients and carriers are described, for example, in “Remingtons Pharmaceutical Sciences” Mack Pub. Co., New Jersey (1991) , in “Remington: The Science and Practice of Pharmacy” , Ed. University of the Sciences in Philadelphia, 21^st Edition, LWW (2005) , which are incorporated herein by reference.

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

In some embodiments, the pharmaceutical compositions of the present disclosure can be formulated so that a dosage of between 0.001-1000 mg/kg body weight/day, for example, 0.01-800 mg/kg body weight/day, 0.01-700 mg/kg body weight/day, 0.01-600 mg/kg body weight/day, 0.01-500 mg/kg body weight/day, 0.01-400 mg/kg body weight/day, 0.01-300 mg/kg body weight/day, 0.1-200 mg/kg body weight/day, 0.1-150 mg/kg body weight/day, 0.1-100 mg/kg body weight/day, 0.5-100 mg/kg body weight/day, 0.5-80 mg/kg body weight/day, 0.5-60 mg/kg body weight/day, 0.5-50 mg/kg body weight/day, 1-50 mg/kg body weight/day, 1-45 mg/kg body weight/day, 1-40 mg/kg body weight/day, 1-35 mg/kg body weight/day, 1-30 mg/kg body weight/day, 1-25 mg/kg body weight/day of the compounds provided herein, or a pharmaceutically acceptable salt thereof, can be administered. In some instances, dosage levels below the lower limit of the aforesaid range may be more than adequate, while in other cases still larger doses may be employed without causing any harmful side effect, provided that such larger doses are first divided into several small doses for administration throughout the day. For further information on routes of administration and dosage regimes, see Chapter 25.3 in Volume 5 of Comprehensive Medicinal Chemistry (Corwin Hansch; Chairman of Editorial Board) , Pergamon Press 1990, which is specifically incorporated herein by reference.

In some embodiments, the pharmaceutical compositions of the present disclosure can be formulated as short-acting, fast-releasing, long-acting, and sustained-releasing. Accordingly, the pharmaceutical formulations of the present disclosure may also be formulated for controlled release or for slow release.

In a further aspect, there is also provided veterinary compositions comprising one or more molecules or compounds of the present disclosure or pharmaceutically acceptable salts thereof and a veterinary carrier. Veterinary carriers are materials useful for the purpose of administering the composition and may be solid, liquid or gaseous materials which are otherwise inert or acceptable in the veterinary art and are compatible with the active ingredient. These veterinary compositions may be administered parenterally, orally or by any other desired route.

The pharmaceutical compositions or veterinary compositions may be packaged in a variety of ways depending upon the method used for administering the drug. For example, an article for distribution can include a container having deposited therein the compositions in an appropriate form. Suitable containers are well known to those skilled in the art and include materials such as bottles (plastic and glass) , sachets, ampoules, plastic bags, metal cylinders, and the like. The container may also include a tamper-proof assemblage to prevent indiscreet access to the contents of the package. In addition, the container has deposited thereon a label that describes the contents of the container. The label may also include appropriate warnings. The compositions may also be packaged in unit-dose or multi-dose containers, for example sealed ampoules and vials, and may be stored in a freeze-dried (lyophilized) condition requiring only the addition of the sterile liquid carrier, for example water, for injection immediately prior to use. Extemporaneous injection solutions and suspensions are prepared from sterile powders, granules and tablets of the kind previously described.

In a further aspect, there is also provided pharmaceutical compositions comprise one or more compounds of the present disclosure, or a pharmaceutically acceptable salt thereof, as a first active ingredient, and a second active ingredient.

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

In some embodiments, the second active ingredient can include:

(i) antiproliferative/antineoplastic drugs and combinations thereof, as used in medical oncology such as alkylating agents (for example cis-platin, carboplatin, cyclophosphamide, nitrogen mustard, melphalan, chlorambucil, busulphan and nitrosoureas) ; antimetabolites (for example antifolates such as fluoropyrimidines like 5-fluorouracil and tegafur, raltitrexed, methotrexate, cytosine arabinoside, hydroxyurea and gemcitabine) ; antitumour antibiotics (for example anthracyclines like adriamycin, bleomycin, doxorubicin, daunomycin, epirubicin, idarubicin, mitomycin-C, dactinomycin and mithramycin) ; antimitotic agents (for example vinca alkaloids like vincristine, vinblastine, vindesine and vinorelbine and taxoids like paclitaxel and taxotere) ; and topoisomerase inhibitors (for example epipodophyllotoxins like etoposide and teniposide, amsacrine, topotecan and camptothecins) ;

(ii) cytostatic agents such as antioestrogens (for example tamoxifen, toremifene, raloxifene, droloxifene and iodoxyfene) , oestrogen receptor down regulators (for example fulvestrant) , antiandrogens (for example bicalutamide, flutamide, nilutamide and cyproterone acetate) , LHRH antagonists or LHRH agonists (for example goserelin, leuprorelin and buserelin) , progestogens (for example megestrol acetate) , aromatase inhibitors (for example as anastrozole, letrozole, vorazole and exemestane) and inhibitors of 5a-reductase such as finasteride;

(iii) anti-invasion agents (for example c-Src kinase family inhibitors like 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} thiazole-5-carboxamide (dasatinib, BMS-354825) , and metalloproteinase inhibitors like marimastat and inhibitors of urokinase plasminogen activator receptor function) ;

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

(v) antiangiogenic agents such as those which inhibit the effects of vascular endothelial growth factor, [for example the anti-vascular endothelial cell growth factor antibody bevacizumab (Avastin^TM) and VEGF receptor tyrosine kinase inhibitors such as 4- (4-bromo-2-fluoroanilino) -6-methoxy-7- (1-methylpiperidin-4-ylmethoxy) quinazoline (ZD6474; Example 2 within WO 01/32651) , 4- (4-fluoro-2-methylindol-5-yloxy) -6-methoxy-7- (3-pyrrolidin-1-ylpropoxy) quinazoline (AZD2171; Example 240 within WO 00/47212) , vatalanib (PTK787; WO 98/35985) and SU11248 (sunitinib; WO 01/60814) , and compounds that work by other mechanisms (for example linomide, inhibitors of integrin ανβ3 function and angiostatin) ] ;

(vi) vascular damaging agents such as combretastatin 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 therapies, such as ISIS 2503, an anti-ras antisense agent;

(viii) gene therapy approaches, including approaches to replace aberrant genes such as aberrant p53 or aberrant BRCAl or BRCA2, GDEPT (gene-directed enzyme pro-drug therapy) approaches such as those using cytosine deaminase, thymidine kinase or a bacterial nitroreductase enzyme and approaches to increase patient tolerance to chemotherapy or radiotherapy such as multi-drug resistance gene therapy; and

(ix) immunotherapeutic approaches, including ex-vivo and in-vivo approaches to increase the immunogenicity of patient tumour cells, such as transfection with cytokines such as interleukin 2, interleukin 4 or granulocyte -macrophage colony stimulating factor, approaches to decrease T-cell anergy, approaches using transfected immune cells such as cytokine-transfected dendritic cells, approaches using cytokine-trtnsfected tumour cell lines and approaches using anti-idiotypic antibodies.

Method of treatment of disease

In an aspect, the present disclosure provides compounds of Formula (I) or pharmaceutically acceptable salts thereof, which are capable of inhibiting ATR kinase. The inhibitory properties of compounds of Formula (I) may be demonstrated using the test procedures set out herein.

Accordingly, the compounds of Formula (I) may be used in the treatment (therapeutic or prophylactic) of conditions or diseases in a subject which are mediated by ATR kinase.

As used herein, a “subject” refers to a human and a non-human animal. Examples of a non-human animal include all vertebrates, e.g., mammals, such as non-human primates (particularly higher primates) , dog, rodent (e.g., mouse or rat) , guinea pig, cat, and non-mammals, such as birds, amphibians, reptiles, etc. In a preferred embodiment, the subject is a human. In another embodiment, the subject is an experimental animal or animal suitable as a disease model.

In some embodiments, the compounds of Formula (I) can be used as anti-tumour agents. In some embodiments, the compounds of Formula (I) can be used as anti-proliferative, apoptotic and/or anti-invasive agents in the containment and/or treatment of solid and/or liquid tumour disease. In certain embodiments, the compounds of Formula (I) are useful in the prevention or treatment of those tumours which are sensitive to inhibition of ATR. In certain embodiments, the compounds of Formula (I) are useful in the prevention or treatment of those tumours which are mediated alone or in part by ATR.

In some embodiments, the compounds of Formula (I) are useful for the treatment of proliferative diseases, including malignant diseases such as cancer as well as non-malignant diseases such as inflammatory diseases, obstructive airways diseases, immune diseases or cardiovascular diseases.

In some embodiments, the compounds of Formula (I) are useful for the treatment of cancer, for example but not limited to, haematologic malignancies such as leukaemia, multiple myeloma, lymphomas such as Hodgkin's disease, non-Hodgkin's lymphomas (including mantle cell lymphoma) , and myelodysplastic syndromes, and also solid tumours and their metastases such as breast cancer, lung cancer (non-small cell lung cancer (NSCL) , small cell lung cancer (SCLC) , squamous cell carcinoma) , endometrial cancer, tumours of the central nervous system such as gliomas, dysembryoplastic neuroepithelial tumour, glioblastoma multiforme, mixed gliomas, medulloblastoma, retinoblastoma, neuroblastoma, germinoma and teratoma, cancers of the gastrointestinal tract such as gastric cancer, oesophagal cancer, hepatocellular (liver) carcinoma, cholangiocarcinomas, colon and rectal carcinomas, cancers of the small intestine, pancreatic cancers, cancers of the skin such as melanomas (in particular metastatic melanoma) , thyroid cancers, cancers of the head and neck and cancers of the salivary glands, prostate, testis, ovary, cervix, uterus, vulva, bladder, kidney (including renal cell carcinoma, clear cell and renal oncocytoma) , squamous cell carcinomas, sarcomas such as osteosarcoma, chondrosarcoma, leiomyosarcoma, soft tissue sarcoma, Ewing's sarcoma, gastrointestinal stromal tumour (GIST) , Kaposi's sarcoma, and paediatric cancers such as rhabdomyosarcomas and neuroblastomas.

In some embodiments, the compounds of Formula (I) are useful for the treatment of autoimmune and/or inflammatory diseases, for example but not limited to, allergy, 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 disease, bullous pemphigoid, coeliac disease, chagas disease, chronic obstructive pulmonary disease, chronic Idiopathic thrombocytopenic purpura (ITP) , churg-strauss syndrome, Crohn's disease, dermatomyositis, diabetes mellitus type 1, endometriosis, Goodpasture's syndrome (and associated glomerulonephritis and pulmonary hemorrhage) , graves' disease, guillain-barre syndrome, hashimoto's disease, hidradenitis suppurativa, idiopathic thrombocytopenic purpura, interstitial cystitis, irritable bowel syndrome, lupus erythematosus, morphea, multiple sclerosis, myasthenia gravis, narcolepsy, neuromyotonia, Parkinson's disease, pemphigus vulgaris, pernicious anaemia, 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 graft rejection and hyperacute rejection of transplanted organs, vasculitis (ANCA-associated and other vasculitides) , vitiligo, and Wegener's granulomatosis.

As used herein, the term “therapy” is intended to have its normal meaning of dealing with a disease in order to entirely or partially relieve one, some or all of its symptoms, or to correct or compensate for the underlying pathology, thereby achieving beneficial or desired clinical results. For purposes of this disclosure, beneficial or desired clinical results include, but are not limited to, alleviation of symptoms, diminishment of extent of disease, stabilized (i.e., not worsening) state of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, and remission (whether partial or total) , whether detectable or undetectable. “Therapy” can also mean prolonging survival as compared to expected survival if not receiving it. Those in need of therapy include those already with the condition or disorder as well as those prone to have the condition or disorder or those in which the condition or disorder is to be prevented. The term “therapy” also encompasses prophylaxis unless there are specific indications to the contrary. The terms “therapeutic” and “therapeutically” should be interpreted in a corresponding manner.

As used herein, the term “prophylaxis” or “prophylactic” is intended to have its normal meaning and includes primary prophylaxis to prevent the development of the disease and secondary prophylaxis whereby the disease has already developed and the patient is temporarily or permanently protected against exacerbation or worsening of the disease or the development of new symptoms associated with the disease.

The term “treatment” is used synonymously with “therapy” . Similarly the term “treat” can be regarded as “applying therapy” where “therapy” is as defined herein.

In a further aspect, the present disclosure provides use of the compound of the present disclosure or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of the present disclosure for use in therapy, for example, for use in therapy associated with ATR kinase.

In a further aspect, the present disclosure provides use of the compound of the present disclosure or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of the present disclosure, in the manufacture of a medicament for treating cancer.

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

In some embodiments, the compounds of Formula (I) can be used further combination with other biologically active ingredients (such as, but not limited to, a second and different antineoplastic agent) and non-drug therapies (such as, but not limited to, surgery or radiation treatment) . For instance, the compounds of Formula (I) can be used in combination with other pharmaceutically active compounds, or non-drug therapies, preferably compounds that are able to enhance the effect of the compounds of Formula (I) . The compounds of Formula (I) can be administered simultaneously (as a single preparation or separate preparation) or sequentially to the other therapies. In general, a combination therapy envisions administration of two or more drugs/treatments during a single cycle or course of therapy.

In some embodiments, the compounds of Formula (I) are used in combination with one or more of traditional chemotherapeutic agents, which encompass a wide range of therapeutic treatments in the field of oncology. These agents are administered at various stages of the disease for the purposes of shrinking tumors, destroying remaining cancer cells left over after surgery, inducing remission, maintaining remission and/or alleviating symptoms relating to the cancer or its treatment.

In some embodiments, the compounds of Formula (I) are used in combination with one or more targeted anti-cancer agents that modulate protein kinases involved in various disease states.

In some embodiments, the compounds of Formula (I) are used in combination with one or more targeted anti-cancer agents that modulate non-kinase biological targets, pathway, or processes.

In some embodiments, the compounds of Formula (I) are used in combination with one or more of other anti-cancer agents that include, but are not limited to, gene therapy, RNAi cancer therapy, chemoprotective agents (e.g., amfostine, mesna, and dexrazoxane) , drug-antibody conjugate (e.g brentuximab vedotin, ibritumomab tioxetan) , cancer immunotherapy such as Interleukin-2, cancer vaccines (e.g., sipuleucel-T) or monoclonal antibodies (e.g., Bevacizumab, Alemtuzumab, Rituximab, Trastuzumab, etc) .

In some embodiments, the compounds of Formula (I) are used in combination with one or more anti-inflammatory agent including but not limited to NSAIDs, non-specific and COX-2 specific cyclooxgenase enzyme inhibitors, gold compounds, corticosteroids, methotrexate, tumor necrosis factor receptor (TNF) receptors antagonists, immunosuppressants and methotrexate.

In some embodiments, the compounds of Formula (I) are used in combination with radiation therapy or surgeries. Radiation is commonly delivered internally (implantation of radioactive material near cancer site) or externally from a machine that employs photon (x-ray or gamma-ray) or particle radiation. Where the combination therapy further comprises radiation treatment, the radiation treatment may be conducted at any suitable time so long as a beneficial effect from the co-action of the combination of the therapeutic agents and radiation treatment is achieved.

Accordingly, in a further aspect, the present disclosure provides a method for treating diseases associated with ATR kinase in a subject in need thereof, comprising administering an effective amount of a compound of the present disclosure or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of the present disclosure to the subject.

EXAMPLES

For the purpose of illustration, the following examples are included. However, it is to be understood that these examples do not limit the present disclosure and are only meant to suggest a method of practicing the present disclosure. Persons skilled in the art will recognize that the chemical reactions described may be readily adapted to prepare a number of other compounds of the present disclosure, and alternative methods for preparing the compounds of the present disclosure are deemed to be within the scope of the present disclosure. For example, the synthesis of non-exemplified compounds according to the present disclosure may be successfully performed by modifications apparent to those skilled in the art, e.g., by appropriately protecting interfering groups, by utilizing other suitable reagents and building blocks known in the art other than those described, and/or by making routine modifications of reaction conditions. Alternatively, other reactions disclosed herein or known in the art will be recognized as having applicability for preparing other compounds of the present disclosure.

Example 1

Synthesis of (R) -4- (7- (3, 5-dimethyl-1H-1, 2, 4-triazol-1-yl) -3- (3-methyl-1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine

Step 1. (3R) -4- (7- (3, 5-dimethyl-1H-1, 2, 4-triazol-1-yl) -3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine

To a solution of 3, 5-dimethyl-4H-1, 2, 4-triazole (45 mg, 0.461 mmol) in dioxane (30 mL) were added (3R) -4- {7-chloro-3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine (100 mg, 0.230 mmol) , Ruphos (12 mg, 0.046 mmol) , RuPhos-Pd-G3 (20 mg, 0.046 mmol) and Cs₂CO₃ (224 mg, 0.690 mmol) . The reaction mixture was degassed and charged with N₂ for several times and then stirred at 100℃ for 16 h under N₂ atmosphere. LC-MS showed the reaction was completed. The reaction mixture was cooled to room temperature and filtered. The filtrate was concentrated in vacuum. The residue was purified by flash chromatography (silica gel, 0 ～ 100 %, ethyl acetate in petroleum ether) to afford (3R) -4- (7- (3, 5-dimethyl-1H-1, 2, 4-triazol-1-yl) -3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine (52 mg, 0.095 mmol, 41.06%) as a colorless oil. LCMS: m/z 495.7 (M+H) ⁺.

Step 2. (R) -4- (7- (3, 5-dimethyl-1H-1, 2, 4-triazol-1-yl) -3- (3-methyl-1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine

To a solution of (3R) -4- (7- (3, 5-dimethyl-1H-1, 2, 4-triazol-1-yl) -3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine (52 mg, 0.105 mmol) in DCM (1.5 mL) was added HCl (4 M in dioxane, 3 mL) . The mixture was stirred at r.t for 0.5 h. LC-MS showed the reaction was completed. The mixture was concentrated in vacuum. The residue was purified by Prep-HPLC (C₁₈, 10-95%, MeCN in H₂O with 0.1%HCOOH) to afford (R) -4- (7- (3, 5-dimethyl-1H-1, 2, 4-triazol-1-yl) -3- (3-methyl-1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine (15 mg, 0.037 mmol, 34.76%) . LCMS: m/z 411.6 (M+H) ⁺. ¹H NMR (400 MHz, DMSO) δ 13.19 (s, 1H) , 7.28 (s, 1H) , 7.12 (s, 1H) , 4.53 (d, J = 5.1 Hz, 1H) , 4.15 (d, J = 12.7 Hz, 1H) , 4.07 (d, J = 8.5 Hz, 1H) , 3.84 (d, J = 11.3 Hz, 1H) , 3.75 (d, J = 11.4 Hz, 1H) , 3.60 (t, J = 10.4 Hz, 1H) , 2.80 (s, 3H) , 2.40 (s, 3H) , 2.32 (s, 3H) , 1.28 (d, J = 6.6 Hz, 3H)

Example 2

Synthesis of (R) -3-methyl-4- (7- (5-methyl-1H-1, 2, 4-triazol-1-yl) -3- (3-methyl-1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) morpholine

(R) -3-methyl-4- (7- (3-methyl-1H-1, 2, 4-triazol-1-yl) -3- (3-methyl-1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) morpholine

Step 1. Mixture of (3R) -3-methyl-4- (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -7- (5-methyl-1H-1, 2, 4-triazol-1-yl) isothiazolo [4, 5-b] pyridin-5- yl) morpholine and (3R) -3-methyl-4- (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -7- (3-methyl-1H-1, 2, 4-triazol-1-yl) isothiazolo [4, 5-b] pyridin-5-yl) morpholine

To a mixture of (3R) -4- {7-chloro-3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine (150 mg, 0.346 mmol) , 5-methyl-1H-1, 2, 4-triazole (57 mg, 0.691 mmol) , RuPhos (32 mg, 0.069 mmol) and Cs₂CO₃ (225 mg, 0.691 mmol) in DMF (8 mL) was added RuPhos-Pd-G3 (27 mg, 0.035 mmol) . The mixture was degassed and charged with N₂ for several times and stirred at 80℃ for 16 h. LC-MS showed the reaction was completed. The mixture was filtered and concentrated to dryness. The residue was purified by flash chromatography (silica gel (12 g) , 0-100%, EA in PE) to give a mixture (120 mg, 72%) of (3R) -3-methyl-4- (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -7- (5-methyl-1H-1, 2, 4-triazol-1-yl) isothiazolo [4, 5-b] pyridin-5-yl) morpholine and (3R) -3-methyl-4- (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -7- (3-methyl-1H-1, 2, 4-triazol-1-yl) isothiazolo [4, 5-b] pyridin-5-yl) morpholine as a yellow oil. LC/MS (ESI) : m/z 481.6 [M+1] ⁺.

Step 2. (R) -3-methyl-4- (7- (5-methyl-1H-1, 2, 4-triazol-1-yl) -3- (3-methyl-1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) morpholine

To a mixture (120 mg, 0.250 mmol) of (3R) -3-methyl-4- (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -7- (5-methyl-1H-1, 2, 4-triazol-1-yl) isothiazolo [4, 5-b] pyridin-5-yl) morpholine and (3R) -3-methyl-4- (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -7- (3-methyl-1H-1, 2, 4-triazol-1-yl) isothiazolo [4, 5-b] pyridin-5-yl) morpholine in DCM (1 mL) was added HCl (4 M in dioxane, 3 mL) . The mixture was stirred at rt for 1 h. LC-MS showed the reaction was completed. The mixture was concentrated to dryness. The crude product was purified by prep-HPLC (C₁₈, 10-95%, MeCN in H₂O with 0.1%HCOOH) to give (R) -3-methyl-4- (7- (5-methyl-1H-1, 2, 4-triazol-1-yl) -3- (3-methyl-1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) morpholine (5 mg, 0.013 mmol, 20.20%) and (R) -3-methyl-4- (7- (3-methyl-1H-1, 2, 4-triazol-1-yl) -3- (3-methyl-1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) morpholine (20 mg, 0.050 mmol, 80.81%) . LC/MS (ESI) m/z: 397.5 [M+1] ⁺. P1: ¹H NMR (400 MHz, DMSO) δ 8.34 (s, 1H) , 7.37 (s, 1H) , 7.13 (s, 1H) , 4.54 (d, J = 4.6 Hz, 1H) , 4.16 (d, J = 13.1 Hz, 1H) , 4.07 (d, J = 8.0 Hz, 1H) , 3.83 (s, 1H) , 3.74 (s, 1H) , 3.60 (s, 1H) , 3.31 (d, J = 3.4 Hz, 1H) , 2.86 (s, 3H) , 2.32 (s, 3H) , 1.29 (d, J = 6.6 Hz, 3H) . P2: ¹H NMR (400 MHz, DMSO) δ 9.74 (d, J = 1.7 Hz, 1H) , 7.72 (d, J = 4.6 Hz, 1H) , 7.12 (s, 1H) , 4.63 –4.54 (m, 1H) , 4.17 (d, J = 13.3 Hz, 1H) , 4.09 –4.03 (m, 1H) , 3.84 (s, 1H) , 3.74 (d, J = 9.0 Hz, 1H) , 3.59 (d, J = 2.3 Hz, 1H) , 3.30 (t, J = 11.3 Hz, 1H) , 2.47 (s, 3H) , 2.32 (s, 3H) , 1.27 (d, J =6.6 Hz, 3H) .

Example 3

Synthesis of (R) -2- (3- (3-methyl-1H-pyrazol-5-yl) -5- (3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) propan-2-amine

Step 1. 3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridine-7-carbonitrile

To a solution of (3R) -4- {7-chloro-3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine (300 mg, 0.691 mmol) in DMF (2 mL) was added Zn (CN) ₂ (162 mg, 1.38 mmol) , DPPF (77 mg, 0.138 mmol) and Pd₂ (dba) ₃ (63 mg, 0.069 mmol) . The reaction was stirred at 120℃ overnight under nitrogen atmosphere. LC-MS showed the reaction was complete. The reaction was diluted with EA (10 mL) and water (10 mL) . The organic layer was separated, washed with brine, dried over anhydrous Na₂SO₄, filtered and concentrated in vacuo. The residue was purified column chromatography on silica gel (PE: EA = 1: 1) to afford the desired product (200 mg, 0.471 mmol, 68.2%) as a yellow solid. LC/MS (ESI) m/z: 425 (M+H) ⁺.

Step 2. 2- (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) propan-2-amine

To a solution of 3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridine-7-carbonitrile (100 mg, 0.236 mmol) in THF (2 mL) were added Methyl magnesium bromide (in ethyl ether) (0.71 mL, 2.12 mmol) drop wise at 0℃. The mixture was stirred at 0℃ for 30 min, Titanium tetraisopropanolate (0.21 mL, 0.707 mmol) was added to the mixture dropwise. Then the mixture was stirred at 50℃ overnight. The reaction was diluted with EA (20 mL) and water (20 mL) . The organic layer was separated, washed with brine, dried over anhydrous Na₂SO₄, filtered and concentrated in vacuo. The residue was purified by prep-TLC (DCM: MeOH = 20: 1, V/V) to afford the desired product (30 mg, 27.9%) as a yellow solid. LC/MS (ESI) m/z: 589 (M+H) ⁺.

Step 3. (R) -2- (3- (3-methyl-1H-pyrazol-5-yl) -5- (3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) propan-2-amine

To a solution of 2- {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} propan-2-amine (30 mg, 0.066 mmol) in DCM (1 mL) and HCl/Dioxane (4M) (1 mL) . The mixture was stirred at room temperature for 30 min. LC-MS showed the reaction was complete. The reaction mixture was concentrated under vacuo. The residue was purified by prep-HPLC (C₁₈, 10-95%, MeOH in H₂O with 0.1%HCOOH) to afford the desired product (3 mg, 12.3%) . LC/MS (ESI) m/z: 372 [M+H] ⁺. ¹H NMR (400 MHz, DMSO-d6) δ8.22 (s, 1H) , 7.07 (s, 1H) , 6.99 (s, 1H) , 4.53 (d, J = 5.4 Hz, 1H) , 4.11 –4.00 (m, 2H) , 3.81 (d, J = 11.3 Hz, 1H) , 3.72 (d, J = 10.8 Hz, 1H) , 3.57 (t, J = 10.2 Hz, 1H) , 3.22 (m, 1H) , 2.29 (s, 3H) , 1.49 (d, J = 3.1 Hz, 6H) , 1.20 (d, J = 6.6 Hz, 3H) .

Example 4

Synthesis of (3R) -4- [7- (1-cyclopropyl-1H-1, 2, 3-triazol-5-yl) -3- (3-methyl-1H-pyrazol-5-yl) - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] -3-methylmorpholine

Step 1. N'- [ (2Z) -1, 1-dimethoxypropan-2-ylidene] -4-methylbenzene-1- sulfonohydrazide

To a mixture of 2, 2-dimethoxyacetaldehyde (1.7 mL, 11.3 mmol) in MeOH (10 mL) was added 4-methylbenzene-1-sulfonohydrazide (2.0 mg, 10.7 mmol) under N₂. The mixture was stirred at rt for 5 min. LC-MS showed the reaction was complete. The mixture was concentrated in vacuo to give crude product N- {2- [2-oxo-5- (tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1, 2-dihydropyridin-1-yl] ethyl} methanesulfonamide (3.0 g, 10.5 mmol) , which was used for the next step without further purification.

Step 2. 1-cyclopropyl-1H-1, 2, 3-triazole

To a mixture of N'- [ (2Z) -1, 1-dimethoxypropan-2-ylidene] -4-methylbenzene-1-sulfonohydrazide (3.0 g, 10.5 mmol) in MeOH (30 mL) was added cyclopropanamine (0.8 mL, 11.5 mmol) and TEA (1.60 mL, 11.5 mmol) under N₂. The mixture was stirred at 75℃ for 15 h. LC-MS showed the reaction was complete. H₂O (10 mL) was added to the reaction mixture. The reaction mixture was extracted with EtOAc (10 mL x 3) . The combined organic layer was washed with brine, dried over Na₂SO₄, filtered and concentrated in vacuo. The residue was purified by flash column chromatography (Silica gel, 0～100%ethyl acetate in petroleum ether) to give 1-cyclopropyl-1H-1, 2, 3-triazole (400 mg, 3.66 mmol, 35%) . LC/MS (ESI) : m/z 110 [M+H] ⁺.

Step 3. (3R) -4- [7- (1-cyclopropyl-1H-1, 2, 3-triazol-5-yl) -3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] -3-methylmorpholine

To a solution of (3R) -4- {7-chloro-3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine (50 mg, 0.115 mmol) in DMA (3 mL) was added 1-cyclopropyl-1H-1, 2, 3-triazole (76 mg, 0.691 mmol) , tetramethylazanium acetate (32 mg, 0.230 mmol) and Bis (triphenylphosphine) palladium (II) chloride (13 mg, 0.016 mmol) . The reaction was stirred at 140℃ under N₂ for 6h. LC-MS showed the reaction was complete. The mixture was poured into H₂O (10 mL) and extracted with EtOAc (10 mL x 3) . The combined organic layer was washed with brine and dried over Na₂SO₄, filtered and concentrated in vacuo. The reaction was purified by prep-TLC (PE: EA=1: 1) to give (3R) -4- [3-chloro-7- (1-cyclopropyl-1H-1, 2, 3-triazol-5-yl) - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] -3-methylmorpholine (15 mg, 0.040 mmol, 49%) as a yellow oil. LC/MS (ESI) : m/z 506.8 [M+H] ⁺.

Step 4. (3R) -4- [7- (1-cyclopropyl-1H-1, 2, 3-triazol-5-yl) -3- (3-methyl-1H-pyrazol-5-yl) - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] -3-methylmorpholine

To a solution of (3R) -4- [7- (1-cyclopropyl-1H-1, 2, 3-triazol-5-yl) -3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] -3- methylmorpholine (15 mg, 0.040 mmol) in DCM (1 mL) was added HCl-Dioxane (2 mL) . The reaction was stirred at room temperature for 1 h. LC-MS showed the reaction was complete. The reaction mixture was concentrated under reduced pressure. The residue was purified by prep-HPLC (C18, 10-95%, MeOH in H₂O with 0.1%HCOOH) to give (3R) -4- [7- (1-cyclopropyl-1H-1, 2, 3-triazol-5-yl) -3- (3-methyl-1H-pyrazol-5-yl) - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] -3-methylmorpholine (5 mg, 0.012 mmol, 9.99%) . LC/MS (ESI) : m/z 422.9 [M+H] ⁺. ¹H NMR (400 MHz, DMSO) δ13.12 (d, J = 121.5 Hz, 1H) , 8.25 (s, 1H) , 7.62 (s, 1H) , 7.15 (s, 1H) , 4.72 –4.40 (m, 1H) , 4.24 –4.00 (m, 3H) , 3.84 –3.68 (m, 2H) , 3.66 –3.54 (m, 1H) , 3.29 –3.24 (m, 1H) , 2.33 (s, 3H) , 1.27 (d, J = 6.5 Hz, 3H) , 1.21 –1.09 (m, 4H) .

Example 5

Synthesis of (R) -4- (7- (1-cyclopropyl-1H-pyrazol-5-yl) -3- (3-methyl-1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine

Step 1. (3R) -4- (7- (1-cyclopropyl-1H-pyrazol-5-yl) -3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine

To a solution of (3R) -4- {7-chloro-3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine (100 mg, 0.230 mmol) in Dioxane (2.0 mL) and H₂O (0.4 mL) were added Pd (PPh₃) ₄ (53 mg, 0.046 mmol) , 1-cyclopropyl-5- (tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole (162 mg, 0.691 mmol) and K₂CO₃ (159 mg, 1.15 mmol) . The mixture was degassed and charged with N₂ for several times and then stirred at 80℃ for 16 h under N₂. LC-MS showed the reaction was completed. The reaction mixture was poured into H₂O (20 mL) and extracted with EA (20 mL*3) . The combined organic phase was washed with brine (20 mL) , dried over anhydrous Na₂SO₄, filtered and concentrated in vacuum. The residue was purified by flash chromatography (silica gel, 0 ～ 100%, ethyl acetate in petroleum ether) to afford (3R) -4- (7- (1-cyclopropyl-1H-pyrazol-5-yl) -3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine (50 mg, 0.099 mmol, 42.91%) as a colorless solid. LC/MS (ESI) : m/z 506.7 [M+H] ⁺.

Step 2. (R) -4- (7- (1-cyclopropyl-1H-pyrazol-5-yl) -3- (3-methyl-1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine

To a solution of (3R) -4- (7- (1-cyclopropyl-1H-pyrazol-5-yl) -3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine (50 mg, 0.099 mmol) in DCM (1.5 mL) was added HCl (4 M in dioxane, 3 mL) . The mixture was stirred at r.t for 0.5 h. LC-MS showed the reaction was completed. The mixture was concentrated in vacuum. The residue was purified by Prep-HPLC (C₁₈, 10-95%, MeOH in H₂O with 0.1%HCOOH) to afford (R) -4- (7- (1-cyclopropyl-1H-pyrazol-5-yl) -3- (3-methyl-1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine (10 mg, 0.024 mmol, 23.99%) as a white solid. LCMS: m/z 422.5 (M+H) ⁺. ¹H NMR (400 MHz, DMSO) δ 13.11 (d, J = 121.4 Hz, 1H) , 7.64 (d, J = 1.7 Hz, 1H) , 7.57 (s, 1H) , 7.15 (s, 1H) , 6.79 (s, 1H) , 4.55 (s, 1H) , 4.17 (d, J = 12.8 Hz, 1H) , 4.06 (d, J = 10.0 Hz, 1H) , 3.96 (d, J = 5.0 Hz, 1H) , 3.82 (d, J = 11.4 Hz, 1H) , 3.74 (d, J = 10.5 Hz, 1H) , 3.59 (t, J = 10.4 Hz, 1H) , 3.27 –3.16 (m, 1H) , 2.32 (s, 3H) , 1.27 (d, J = 6.6 Hz, 3H) , 1.08 –0.92 (m, 4H) .

Example 6

Synthesis of (R) -4- (7- (1-cyclopropyl-1H-1, 2, 3-triazol-5-yl) -3- (1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine

Step 1. (3R) -4- (7- (1-cyclopropyl-1H-1, 2, 3-triazol-5-yl) -3- (1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine

To a solution of (3R) -4- {7-chloro-3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine (100 mg, 0.239 mmol) in DMA (3 mL) were added 4- (difluoromethyl) -4H-1, 2, 4-triazole (156 mg, 1.43 mmol) , Bis(triphenylphosphine) palladium (II) chloride (33 mg, 0.048 mmol) and tetramethylazanium acetate (318 mg, 2.39 mmol) . The mixture was degassed and charged with N₂ for several times and then stirred at 140℃ for 6 h under N₂. LC-MS showed the reaction was completed. The reaction mixture was poured into water (20 mL) and extracted with EA (20 mL *3) . The combined organic layer was washed with brine (20 mL) , dried over Na₂SO₄, filtered and concentrated. The residue was purified by Prep-TLC (PE: EA=1: 1) to give (3R) -4- (7- (1-cyclopropyl-1H-1, 2, 3-triazol-5-yl) -3- (1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine (25 mg, 0.051 mmol, 21.1%) as a yellow solid. LC/MS (ESI) : m/z 493.1 [M+H] ⁺.

Step 2. (R) -4- (7- (1-cyclopropyl-1H-1, 2, 3-triazol-5-yl) -3- (1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine

To a solution of (3R) -4- (7- (1-cyclopropyl-1H-1, 2, 3-triazol-5-yl) -3- (1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine (25 mg, 0.051 mmol) in DCM (1 mL) was added HCl (4 M in dioxane, 2 mL) dropwise. The reaction was stirred at room temperature for 1 h. LC-MS showed the reaction was completed. The reaction mixture was concentrated. The residue was purified by Prep-HPLC (C₁₈, 10-95%, MeOH in H₂O with 0.1% HCOOH) to give (R) -4- (7- (1-cyclopropyl-1H-1, 2, 3-triazol-5-yl) -3- (1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine (4 mg, 0.010 mmol, 19.29%) . LC/MS (ESI) : m/z 408.9 [M+H] ⁺. ¹H NMR (400 MHz, DMSO) δ 13.54 (d, J = 174.5 Hz, 1H) , 8.26 (s, 1H) , 7.68 (d, J = 29.0 Hz, 2H) , 7.43 (s, 1H) , 4.56 (s, 1H) , 4.23 –4.04 (m, 3H) , 3.83 –3.72 (m, 2H) , 3.57 (s, 1H) , 3.13 –2.98 (m, 1H) , 1.32 –1.24 (m, 4H) , 1.15 (dd, J = 8.5, 5.6 Hz, 3H) .

Example 7

Synthesis of (R) -2- (3- (3-methyl-1H-pyrazol-5-yl) -5- (3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) propan-1, 1, 1, 3, 3, 3-d6-2-ol

Step 1. 2- (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) propan-1, 1, 1, 3, 3, 3-d6-2-ol

To a solution of methyl 3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridine-7-carboxylate (100 mg, 0.219 mmol) in THF (5 mL) was added CD₃MgI (111 mg, 0.657 mmol) under N₂ atmosphere at 0℃. The reaction was stirred at 0℃ for 1 hr. LC-MS showed the reaction was complete. The mixture was poured into saturated NH₄Cl and extracted with EA (20 mL *3) . The combined organic layer was washed with brine, dried over anhydrous Na₂SO₄, filtered and concentrated. The residue was purified by flash chromatography (silica gel, 0 ～ 50 %, ethyl acetate in petroleum ether) to afford 2- (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) propan-1, 1, 1, 3, 3, 3-d6-2-ol (85 mg, 0.183 mmol, 83.89%) as a colorless sticky oil. LC/MS (ESI) : m/z 380 (M+H) ⁺.

Step 2. (R) -2- (3- (3-methyl-1H-pyrazol-5-yl) -5- (3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) propan-1, 1, 1, 3, 3, 3-d6-2-ol

To a solution of 2- (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) propan-1, 1, 1, 3, 3, 3-d6-2-ol (50 mg, 0.099 mmol) in DCM (3 mL) was added HCl (4M in dioxane, 3 mL) . The mixture was stirred rt for 0.5h. LC-MS showed the reaction was complete. The mixture was concentrated in vacuum. The residue was purified by prep-HPLC (C₁₈, 10-95%, MeOH in H₂O with 0.1%HCOOH) to afford (R) -2- (3- (3-methyl-1H-pyrazol-5-yl) -5- (3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) propan-1, 1, 1, 3, 3, 3-d6-2-ol (10 mg, 24.43%) . LCMS (ESI) : m/z 380.2 (M+H) ⁺. ¹H NMR (400 MHz, DMSO-d6) δ 7.09 (s, 1H) , 6.56 (s, 1H) , 4.36 (d, J = 4.8 Hz, 1H) , 4.17 (dd, J = 12.6, 2.6 Hz, 2H) , 4.11-3.95 (m, 2H) , 3.83-3.62 (m, 6H) , 3.55 (td, J = 11.8, 2.8 Hz, 3H) , 3.27 –3.00 (m, 2H) , 1.21 (d, J = 6.7 Hz, 3H) .

Example 8

Synthesis of 2- (^2H₃) methyl-2- [3- (3-methyl-1H-pyrazol-5-yl) -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl] (^2H₃) propanenitrile

Step 1. 2- {3-chloro-5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} -2- (^2H₃) methyl (^2H₃) propanenitrilee

To a solution of 2- {3-chloro-5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} acetonitrile (120 mg, 0.389 mmol) and (tert-butoxy) sodium (75 mg, 0.777 mmol) in anhydrous THF (3 mL) at 0℃ was added a solution CD₃I (0.05 mL, 0.777 mmol) in anhydrous THF (2mL) dropwise. The mixture was stirred at room temperature for 1 h. LC-MS showed the reaction was complete. The reaction mixture was diluted with EA (40 mL) , then washed with water and brine, dried over anhydrous Na₂SO₄, filtered and concentrated. The residue was purified by column chromatography on silica gel (PE: EA = 2: 1, V/V) to afford 2- {3-chloro-5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} -2- (^2H₃) methyl (^2H₃) propanenitrile (100 mg, 0.292 mmol, 75.05%) as a yellow solid. LC/MS (ESI) : m/z 343.1 [M+H] ⁺.

Step 2. 2- (methyl-d3) -2- (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) propanenitrile-3, 3, 3-d3

To a mixture of (R) -2- (3-chloro-5- (3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) -2- (methyl-d3) propanenitrile-3, 3, 3-d3 (55 mg, 0.160 mmol) , [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] boronic acid (67 mg, 0.321 mmol) and K₂CO₃ (66.51 mg, 0.481 mmol) in dioxane (4 mL) and H₂O (0.8 mL) was added Pd (PPh₃) ₄ (37 mg, 0.032 mmol) . The mixture was stirred at 100℃ for 16 h under N₂ atmosphere. LC-MS showed the reaction was complete. The reaction mixture was diluted with H₂O (20 mL) , then extracted with EA (20 mL×3) . The combined organic layer was washed with brine, dried over anhydrous Na₂SO₄, filtered and concentrated. The residue was purified by column chromatography on silica gel (DCM: MeOH = 20: 1, ) to give 2- (methyl-d3) -2- (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) propanenitrile-3, 3, 3-d3 (30 mg, 39.6%) as a brown solid. LC/MS (ESI) : m/z 473.1 [M+H] ⁺.

Step 3. (R) -2- (methyl-d3) -2- (3- (3-methyl-1H-pyrazol-5-yl) -5- (3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) propanenitrile-3, 3, 3-d3

To a solution of 22- (methyl-d3) -2- (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) propanenitrile-3, 3, 3-d3 (20 mg, 0.042 mmol) in DCM (1 mL) was added HCl-dioxane (4M, 2 mL) dropwise. The reaction was stirred at room temperature for 1 h. LC-MS showed the reaction was complete. The reaction mixture was concentrated under reduced pressure. The residue was purified by prep-HPLC (C₁₈, 10-95%, MeOH in H₂O with 0.1%HCOOH) to give (R) -2- (methyl-d3) -2- (3- (3-methyl-1H-pyrazol-5-yl) -5- (3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) propanenitrile-3, 3, 3-d3 (4 mg, 24.33%) . LC/MS (ESI) : m/z 390.0 [M+H] ⁺. ¹H NMR (400 MHz, DMSO-d6) δ 7.13 (d, J = 8.0 Hz, 2H) , 4.55 (s, 1H) , 4.08 (dd, J = 27.8, 11.7 Hz, 2H) , 3.83 (d, J = 11.1 Hz, 1H) , 3.71 (d, J = 11.1 Hz, 1H) , 3.54 (dd, J = 27.1, 16.0 Hz, 2H) , 2.31 (s, 3H) , 1.25 (d, J = 6.4 Hz, 3H) .

Example 9

Synthesis of (R) -3-methyl-4- (3- (3-methyl-1H-pyrazol-5-yl) -7- (2H-1, 2, 3-triazol-2-yl) isothiazolo [4, 5-b] pyridin-5-yl) morpholine and (R) -3-methyl-4- (3- (3-methyl- 1H-pyrazol-5-yl) -7- (1H-1, 2, 3-triazol-1-yl) isothiazolo [4, 5-b] pyridin-5-yl) morpholine

Step 1. (3R) -3-methyl-4- (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -7- (2H-1, 2, 3-triazol-2-yl) isothiazolo [4, 5-b] pyridin-5-yl) morpholine and (3R) -3-methyl-4- (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -7- (1H-1, 2, 3-triazol-1-yl) isothiazolo [4, 5-b] pyridin-5-yl) morpholine

To a solution of (3R) -4- {7-chloro-3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine (120 mg, 0.277 mmol) in DMA (3 mL) was added 1H-1, 2, 3-triazole (0.05 mL, 0.830 mmol) , CS₂CO₃ (360 mg, 1.106 mmol) , Ruphos (26 mg, 0.055 mmol) and Ruphos Pd G₃ (46 mg, 0.055 mmol) . After the reaction was stirred at 100℃ overnight under nitrogen atmosphere. LC-MS showed the reaction was complete. The reaction was diluted with EA (20 mL) and water (20 mL) . The organic layer was separated, washed with brine, dried over anhydrous Na₂SO₄, filtered and concentrated in vacuo. The residue was purified by Pre-TLC (DMC: MeOH = 40: 1, V/V) to afford 9-4 (40 mg, 0.086 mmol, 31.0%) as a yellow solid and 9-3 (60 mg, 0.129 mmol, 46.5%) as a yellow solid. LC/MS (ESI) m/z: 467 (M+H) ⁺.

Step 2. (R) -3-methyl-4- (3- (3-methyl-1H-pyrazol-5-yl) -7- (2H-1, 2, 3-triazol-2-yl) isothiazolo [4, 5-b] pyridin-5-yl) morpholine

To a solution of (3R) -3-methyl-4- (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -7- (2H-1, 2, 3-triazol-2-yl) isothiazolo [4, 5-b] pyridin-5-yl) morpholine (40 mg, 0.086 mmol) in DCM (1 mL) and HCl/Dioxane (4M) (1 mL) . The mixture was stirred at room temperature for 1 h. LC-MS showed the reaction was complete. The reaction mixture was concentrated under vacuo. The residue was purified by Pre-HPLC (C₁₈, 10-95%, MeOH in H₂O with 0.1%HCOOH) to afford the desired product (20 mg, 0.052 mmol, 61.0%) . LC/MS (ESI) m/z: 383 [M+H] ⁺.

¹HNMR (400 MHz, DMSO) δ 13.10 (d, J = 114.2 Hz, 1H) , 8.43 (s, 2H) , 7.70 (s, 1H) , 7.13 (s, 1H) , 4.56 (d, J = 5.1 Hz, 1H) , 4.16 (d, J = 12.7 Hz, 1H) , 4.06 (d, J = 8.6 Hz, 1H) , 3.83 (d, J = 11.3 Hz, 1H) , 3.75 (d, J =9.0 Hz, 1H) , 3.60 (t, J = 10.4 Hz, 1H) , 3.29 (m, 1H) , 2.32 (s, 3H) , 1.27 (d, J = 6.6 Hz, 3H) .

Step 3. (R) -3-methyl-4- (3- (3-methyl-1H-pyrazol-5-yl) -7- (1H-1, 2, 3-triazol-1-yl) isothiazolo [4, 5-b] pyridin-5-yl) morpholine

To a solution of (3R) -3-methyl-4- (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -7- (1H-1, 2, 3-triazol-1-yl) isothiazolo [4, 5-b] pyridin-5-yl) morpholine (60 mg, 0.129 mmol) in DCM (1 mL) and HCl/Dioxane (4M) (1 mL) . The mixture was stirred at room temperature for 1 h. LC-MS showed the reaction was complete. The reaction mixture was concentrated under vacuo. The residue was purified by Pre-HPLC (C₁₈, 10-95%, MeOH in H₂O with 0.1%HCOOH) to afford the desired product (30 mg, 0.078 mmol, 61.0%) . LC/MS (ESI) m/z: 383 [M+H] ⁺.

¹HNMR (400 MHz, DMSO) δ 13.22 (s, 1H) , 9.48 (s, 1H) , 8.19 (s, 1H) , 7.78 (s, 1H) , 7.14 (s, 1H) , 4.63 (d, J = 7.3 Hz, 1H) , 4.21 (d, J = 12.7 Hz, 1H) , 4.08 (d, J = 10.8 Hz, 1H) , 3.87 (d, J = 11.2 Hz, 1H) , 3.76 (d, J =11.2 Hz, 1H) , 3.59 (d, J = 11.8 Hz, 1H) , 3.30 (s, 1H) , 2.32 (s, 3H) , 1.29 (d, J = 6.5 Hz, 3H) .

Example 10

Synthesis of (R) -1- (3- (3-methyl-1H-pyrazol-5-yl) -5- (3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) cyclopropan-1-ol

Step 1. 1- (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) cyclopropan-1-ol

To a solution of 3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridine-7-carbonitrile (80 mg, 0.188 mmol) in THF (2 mL) were added Ethyl magnesium bromide (0.940 mL, 0.940 mmol) drop wise at 0℃. The mixture was stirred at 0℃ for 30 min. Titanium tetraisopropanolate (0.059 mL, 0.564 mmol) was added to the mixture dropwise. Then the mixture was stirred at 50℃ overnight. The reaction was diluted with EA (20 mL) and water (20 mL) . The organic layer was separated, washed with brine, dried over anhydrous Na₂SO₄, filtered and concentrated in vacuo. The residue was purified by Pre-TLC (DCM: MeOH = 20: 1, V/V) to afford the desired product (20 mg, 0.044 mmol, 23.4%) . LC/MS (ESI) m/z: 456 (M+H) ⁺.

Step 2. (R) -1- (3- (3-methyl-1H-pyrazol-5-yl) -5- (3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) cyclopropan-1-ol

To a solution of 1- (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) cyclopropan-1-ol (20 mg, 0.044 mmol) in DCM (1 mL) and HCl/Dioxane (4M) (1 mL) . The mixture was stirred at room temperature for 30 min. LC-MS showed the reaction was complete. The reaction mixture was concentrated under vacuo. The residue was purified by prep-HPLC (C₁₈, 10-95%, MeOH in H₂O with 0.1%HCOOH) to afford the desired product (5 mg, 0.013 mmol, 30.7%) . LC/MS (ESI) m/z: 372 [M+H] ⁺. ¹H NMR (400 MHz, DMSO) δ 13.07 (NH, 1H) , 7.87 (s, 1H) , 7.11 (s, 1H) , 4.66 (s, 1H) , 4.21 (d, J =13.5 Hz, 1H) , 4.07 (d, J = 9.3 Hz, 1H) , 3.85 (d, J = 11.6 Hz, 1H) , 3.75 (d, J = 9.7 Hz, 1H) , 3.60 (t, J =10.9 Hz, 1H) , 3.29 (m, 1H) , 2.31 (s, 3H) , 1.27 (d, J = 6.5 Hz, 3H) , 1.18 (t, J = 7.0 Hz, 4H) .

Example 11

Synthesis of (R) -4- (7- (4- (difluoromethyl) -4H-1, 2, 4-triazol-3-yl) -3- (3-methyl-1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine

Step 1. 4- (difluoromethyl) -4H-1, 2, 4-triazole

To a solution of 4H-1, 2, 4-triazole (13.0 mL, 217 mmol) in THF (250 mL) were added NaH (17.4 g, 434 mmol) and diethyl (bromodifluoromethyl) phosphonate (116 g, 434 mmol) at 0℃. The mixture was stirred at 0℃ for 3 h. TLC showed the reaction was complete. The mixture was quenched with H₂O (10 mL) slowly, then diluted with H₂O (250 mL) and extracted with EtOAc (250 mL *3) . The combined organic layer was washed with brine (250 mL) , dried over Na₂SO₄, filtered and concentrated. The residue was purified by flash chromatography (silica gel (120 g) , 0-10%, MeOH in DCM) to give 4- (difluoromethyl) -4H-1, 2, 4-triazole (400 mg, 3.19 mmol, 1.47%) as a colorless oil. LC/MS (ESI) : m/z 120.1 [M+H] ⁺.

Step 2. (3R) -4- (7- (4- (difluoromethyl) -4H-1, 2, 4-triazol-3-yl) -3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine

To a solution of (3R) -4- {7-chloro-3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine (200 mg, 0.461 mmol) in DMA (3 mL) were added 4- (difluoromethyl) -4H-1, 2, 4-triazole (329 mg, 2.76 mmol) , tetramethylazanium acetate (184 mg, 1.38 mmol) and Bis (triphenylphosphine) palladium (II) chloride (72 mg, 0.092 mmol) . The reaction was degassed and charged with N₂ for several times and then stirred at 140℃ under N₂ for 6 h. LC-MS showed the reaction was completed. The reaction was poured into water (20 mL) and extracted with EA (20mL *3) . The combined organic layer was washed with brine (20 mL) , dried over Na₂SO₄, filtered and concentrated. The residue was purified by Prep-TLC (PE: EA=1: 2) to give (3R) -4- (7- (4- (difluoromethyl) -4H-1, 2, 4-triazol-3-yl) -3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine (50 mg, 0.097 mmol, 21.0%) as a yellow oil. LC/MS (ESI) : m/z 516.9 [M+H] ⁺.

Step 3. (R) -4- (7- (4- (difluoromethyl) -4H-1, 2, 4-triazol-3-yl) -3- (3-methyl-1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine

To a solution of (3R) -4- (7- (4- (difluoromethyl) -4H-1, 2, 4-triazol-3-yl) -3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) - 3-methylmorpholine (50 mg, 0.097 mmol) in DCM (1 mL) was added HCl (4 M in dioxane, 2 mL) . The reaction was stirred at room temperature for 1 h. LC-MS showed the reaction was completed. The reaction mixture was concentrated under reduced pressure. The residue was purified by Prep-HPLC (C₁₈, 10-95%, MeOH in H₂O with 0.1%HCOOH) to give (R) -4- (7- (4- (difluoromethyl) -4H-1, 2, 4-triazol-3-yl) -3- (3- methyl-1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine (10 mg, 0.023 mmol, 23.89%) . LC/MS (ESI) : m/z 432.8 [M+H] ⁺. ¹H NMR (400 MHz, DMSO) δ 9.36 (s, 1H) , 8.28 (t, J = 58.7 Hz, 1H) , 7.37 (s, 1H) , 7.13 (s, 1H) , 4.49 (d, J = 4.8 Hz, 1H) , 4.17 –4.06 (m, 2H) , 3.86 (d, J = 11.4 Hz, 1H) , 3.75 (d, J = 2.9 Hz, 1H) , 3.65 –3.59 (m, 2H) , 3.22 –3.19 (m, 1H) , 2.32 (s, 3H) , 1.27 (d, J = 6.7 Hz, 3H) .

Example 12

Synthesis of (3R) -3-methyl-4- [3- (3-methyl-1H-pyrazol-5-yl) -7- [5- (trifluoromethyl) -1H-1, 2, 3-triazol-1-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] morpholine

Step 1. (3R) -3-methyl-4- {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -7- [5- (trifluoromethyl) -1H-1, 2, 3-triazol-1-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} morpholine

To a solution of (3R) -4- {7-chloro-3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine (100 mg, 0.230 mmol ) in DMA (1.5 mL ) were added 5- (trifluoromethyl) -1H-1, 2, 3-triazole (32 mg, 0.230 mmol ) and Cs₂CO₃ (75 mg, 0.230 mmol) . The reaction was stirred at 160℃ for 5 hours. LC-MS showed the reaction was complete. The mixture was poured into H₂O (10 mL) and extracted with EtOAc (10 mL *3) . The combined organic layer was washed with brine, dried over Na₂SO₄, filtered and concentrated in vacuo. The residue was purified by flash column chromatography (Silica gel, 0～100%ethyl acetate in petroleum ether) to afford (3R) -3-methyl-4- {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -7- [5- (trifluoromethyl) -1H-1, 2, 3-triazol-1-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} morpholine (20 mg, 0.037 mmol, 16.2%) as white solid. LC/MS (ESI) m/z: 535 (M+H) ⁺.

Step 2. (3R) -3-methyl-4- [3- (3-methyl-1H-pyrazol-5-yl) -7- [5- (trifluoromethyl) -1H-1, 2, 3-triazol-1-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] morpholine

To a solution of (3R) -3-methyl-4- {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -7- [5- (trifluoromethyl) -1H-1, 2, 3-triazol-1-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} morpholine (20 mg, 0.037 mmol) in DCM (2 mL) were added HCl (4M in dioxane, 2 mL) . The reaction was stirred at room temperature for 1h. LC-MS showed the reaction was complete. The reaction mixture was concentrated under reduced pressure. The residue was purified by prep-HPLC (C₁₈, 10-95%, MeOH in H₂O with 0.1%HCOOH) to give (3R) -3-methyl-4- [3- (3-methyl-1H-pyrazol-5-yl) -7- [5- (trifluoromethyl) -1H-1, 2, 3-triazol-1-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] morpholine (2 mg, 0.004 mmol, 11.9%) . LC/MS (ESI) m/z: 451 (M+H) ⁺. ¹H NMR (400 MHz, DMSO-d6) δ 6.95 (s, 2H) , 4.33 (d, J = 5.4 Hz, 1H) , 4.10 –4.00 (m, 1H) , 3.95 (d, J =12.2 Hz, 1H) , 3.83 (d, J = 11.3 Hz, 1H) , 3.79 –3.72 (m, 1H) , 3.62 (t, J = 10.3 Hz, 2H) , 2.32 (s, 3H) , 1.29 (d, J = 6.6 Hz, 3H) .

Example 13

Synthesis of (R) -3-methyl-4- (3- (3-methyl-1H-pyrazol-5-yl) -7- (3- (trifluoromethyl) -1H-1, 2, 4-triazol-1-yl) isothiazolo [4, 5-b] pyridin-5-yl) morpholine

Step 1. (3R) -3-methyl-4- (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -7- (3- (trifluoromethyl) -1H-1, 2, 4-triazol-1-yl) isothiazolo [4, 5-b] pyridin-5-yl) morpholine

To a solution of (3R) -4- {7-chloro-3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine (200 mg, 0.461 mmol) in DMA (3 mL) were added 3- (trifluoromethyl) -4H-1, 2, 4-triazole (126 mg, 0.922 mmol) and Cs₂CO₃ (300 mg, 2.30 mmol) . The mixture was stirred at 140℃ for 12 h. LC-MS showed the reaction was completed. The reaction mixture was poured into H₂O (20 mL) and extracted with EA (20 mL *3) . The combined organic phase was washed with brine (20 mL) , dried over anhydrous Na₂SO₄, filtered and concentrated in vacuum. The residue was purified by flash chromatography (silica gel, 0 ～ 100%, ethyl acetate in petroleum ether) to afford (3R) -3-methyl-4- (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -7- (3- (trifluoromethyl) -1H-1, 2, 4-triazol-1-yl) isothiazolo [4, 5-b] pyridin-5-yl) morpholine (230 mg, 0.430 mmol, 93.36%) as a yellow solid. LC/MS (ESI) : m/z 535.6 [M+H] ⁺.

Step 2. (R) -3-methyl-4- (3- (3-methyl-1H-pyrazol-5-yl) -7- (3- (trifluoromethyl) -1H-1, 2, 4-triazol-1-yl) isothiazolo [4, 5-b] pyridin-5-yl) morpholine

To a solution of (3R) -3-methyl-4- (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -7- (3- (trifluoromethyl) -1H-1, 2, 4-triazol-1-yl) isothiazolo [4, 5-b] pyridin-5-yl) morpholine (230 mg, 0.430 mmol) in DCM (3 mL) was added HCl (4 M in dioxane, 3 mL) . The mixture was stirred at r.t for 0.5 h. LC-MS showed the reaction was completed. The mixture was concentrated in vacuum. The residue was purified by Prep-HPLC (C₁₈, 10-95%, MeOH in H₂O with 0.1%HCOOH) to afford (R) -3-methyl-4- (3- (3-methyl-1H-pyrazol-5-yl) -7- (3- (trifluoromethyl) -1H-1, 2, 4-triazol-1-yl) isothiazolo [4, 5-b] pyridin-5-yl) morpholine (10 mg, 0.022 mmol) . LC/MS (ESI) : m/z 451.4 [M+H] ⁺. ¹H NMR (400 MHz, DMSO) δ 13.11 (d, J = 124.8 Hz, 1H) , 10.15 (s, 1H) , 7.90 (s, 1H) , 7.13 (s, 1H) , 4.60 (s, 1H) , 4.20 (d, J = 12.0 Hz, 1H) , 4.09 (d, J = 8.8 Hz, 1H) , 3.88 (d, J = 11.2 Hz, 1H) , 3.75 (d, J = 11.7 Hz, 3H) , 2.32 (s, 3H) , 1.29 (d, J = 6.6 Hz, 3H) .

Example 14

Synthesis of (R) -4- ( (3- (3-methyl-1H-pyrazol-5-yl) -5- (3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) -1, 4l6-oxathiane 4-oxide

Step 1. 4- ( (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3- methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) -1, 4l6-oxathiane 4-oxide

To a solution of (3R) -4- {7-chloro-3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine (100 mg, 0.230 mmol) in dioxane (3 mL) was added (dicyclopropylimino-λ^6-sulfanyl) one (47 mg, 0.325 mmol) , Pb₂ (dba) ₃ (21 mg, 0.023 mmol) , Xantphos (27 mg, 0.046 mmol) and Cs₂CO₃ (250 mg, 0.767 mmol) . The reaction was stirred at 100℃ overnight under nitrogen atmosphere. LC-MS showed the reaction was complete. The reaction solution was filtered and concentrated in vacuo. The residue was purified by flash column chromatography (silica gel, 0～100%, EA in PE) to afford 4- ( (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) -1, 4l6-oxathiane 4-oxide (85 mg, 0.160 mmol, 69.38%) as a yellow solid. LC/MS (ESI) : m/z 533.2 (M+H) ⁺.

Step 2. (R) -4- ( (3- (3-methyl-1H-pyrazol-5-yl) -5- (3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) -1, 4l6-oxathiane 4-oxide

To a solution of 4- ( (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) -1, 4l6-oxathiane 4-oxide (50 mg, 0.094 mmol) DCM (3 mL) was added HCl (4M in dioxane, 3 mL) . The mixture was stirred at rt for 0.5h. LC-MS showed the reaction was complete. The mixture was concentrated in vacuum. The residue was purified by prep-HPLC (C₁₈, 10-95%, MeOH in H₂O with 0.1%HCOOH) to afford 4- { [3- (3-methyl-1H-pyrazol-5-yl) -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl] imino} -1, 4λ^6-oxathian-4-one (15 mg, 0.033 mmol, 22.27%) . LCMS (ESI) : m/z 448.8 (M+H) +. ¹H NMR (400 MHz, DMSO-d6) δ 7.09 (s, 1H) , 6.56 (s, 1H) , 4.36 (d, J = 4.8 Hz, 1H) , 4.17 (dd, J = 12.6, 2.6 Hz, 2H) , 4.11-3.95 (m, 2H) , 3.83-3.62 (m, 6H) , 3.55 (td, J =11.8, 2.8 Hz, 3H) , 3.27-3.00 (m, 2H) , 1.21 (d, J = 6.7 Hz, 3H) .

Example 15

Synthesis of (R) -dimethyl ( (3- (3-methyl-1H-pyrazol-5-yl) -5- (3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) -l6-sulfanone

Step 1. dimethyl ( (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) -l6-sulfanone

To a solution of (3R) -4- {7-chloro-3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine (2 g, 4.61 mmol) in dioxane (25 mL) was added iminodimethyl-λ^6-sulfanone (0.52 g, 5.53 mmol) , CS₂CO₃ (4.50 g, 13.8 mmol) , Pd₂ (dba) ₃ (0.42 g, 0.461 mmol) and Xantphos (0.53 g, 0.922 mmol) . The reaction was stirred at 100℃ for 16h under nitrogen atmosphere. LC-MS showed the reaction was complete. The reaction was poured into H₂O and extracted with EA (3*50 mL) . The organic phase was washed with brine, dried over anhydrous Na₂SO₄, filtered and concentrated in vacuo. The residue was purified by flash column chromatography (Silica gel, 0～100%ethyl acetate in petroleum ether) to give dimethyl ( (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) -l6-sulfanone (1.9 g, 3.87 mmol, 84%) as a yellow solid. LC/MS (ESI) : m/z 491 (M+H) ⁺.

Step 2. (R) -dimethyl ( (3- (3-methyl-1H-pyrazol-5-yl) -5- (3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) -l6-sulfanone

To a solution of dimethyl ( (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) -l6-sulfanone (1.9 g, 3.87 mmol) in DCM (20 mL) was added HCl (4M in dioxane, 20 mL) . The mixture was stirred at room temperature for 1h. LC-MS showed the reaction was complete. The reaction was concentrated in vacuo to give crude product, which was purified by prep-HPLC (C₁₈, 10-95%, MeCN in H₂O with 0.1%CHOOH) to afford (R) -dimethyl ( (3- (3-methyl-1H-pyrazol-5-yl) -5- (3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) -l6-sulfanone (1.07 g, 2.63 mmol, 68%) . LC/MS (ESI) : m/z 407 (M+H) ⁺. ¹H NMR (400 MHz, DMSO-d6) δ 7.06 (s, 1H) , 6.54 (s, 1H) , 4.36 (d, J = 4.7 Hz, 1H) , 4.02 (d, J = 8.1 Hz, 1H) , 3.95 (d, J = 11.3 Hz, 1H) , 3.80 (d, J = 11.3 Hz, 1H) , 3.75 –3.68 (m, 1H) , 3.56 (dd, J = 13.0, 10.4 Hz, 1H) , 3.49 (d, J = 1.9 Hz, 6H) , 3.22 (dd, J = 12.5, 8.8 Hz, 1H) , 2.30 (s, 3H) , 1.22 (d, J = 6.6 Hz, 3H) .

Example 16

Synthesis of (4- [3- (3-methyl-1H-pyrazol-5-yl) -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl] oxan-4-ol

Step 1. (3R) -4- [7- (3, 6-dihydro-2H-pyran-4-yl) -3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] -3-methylmorpholine

To a solution of (3R) -4- {7-chloro-3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine (100 mg, 0.230 mmol) in Dioxane (5 mL) and H₂O (1 mL) was added 2- (3, 6-dihydro-2H-pyran-4-yl) -4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolane (73 mg, 0.346 mmol) , K₂CO₃ (96 mg, 0.691 mmol) and Pd (PPh₃) ₄ (53 mg, 0.046 mmol) . The reaction was stirred at 100℃ for 16h under N₂. LC-MS showed the reaction was complete. H₂O (10 mL) was added to the mixture and the reaction was extracted with EtOAc (3*20 mL) . The combined organic phase was washed with brine, dried over Na₂SO₄, filtered and concentrated in vacuo. The residue was purified by flash column chromatography (Silica gel, 0～100%ethyl acetate in petroleum ether) to give (3R) -4- [7- (3, 6-dihydro-2H-pyran-4-yl) -3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] -3-methylmorpholine (80 mg, 0.166 mmol, 72.1%) as a yellow oil. LC/MS (ESI) : m/z 483.2 [M+H] ⁺.

Step 2. 4- {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} oxan-4-ol

To a solution of (3R) -4- [7- (3, 6-dihydro-2H-pyran-4-yl) -3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] -3-methylmorpholine (100 mg, 0.208 mmol) in IPA (5 mL) and DCM (1 mL) was added phenylsilane (0.052 mL, 0.415 mmol) and Mn (TMHD) ₃ (13 mg, 0.021 mmol) at 0 ℃. The mixture was stirred at rt for 3h under O₂. LC-MS showed the reaction was complete. The mixture was poured into H₂O and extracted with EtOAc (3*20 mL) , washed with brine, dried over Na₂SO₄, filtered and concentrated in vacuo. The residue was purified by prep-TLC (PE: EA=1: 1) to give 4- {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} oxan-4-ol (30 mg, 0.060 mmol, 28.8%) as a white solid. LC/MS (ESI) : m/z 500.1 [M+H] ⁺.

Step 3. 4- [3- (3-methyl-1H-pyrazol-5-yl) -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl] oxan-4-ol

To a solution of 4- {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} oxan-4-ol (30 mg, 0.060 mmol) in DCM (3 mL) was added HCl (4M in dioxane, 1 mL) . The mixture was stirred at rt for 1 hour. LC-MS showed the reaction was complete. The reaction mixture was concentrated. The residue was purified by prep-HPLC (C₁₈, 10-95%, MeOH in H₂O with 0.1%HCOOH) to give 4- [3- (3-methyl-1H-pyrazol-5-yl) -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl] oxan-4-ol (5.8 mg, 0.013 mmol, 22.1%) . LC/MS (ESI) : m/z 416.0 [M+H] ⁺. ¹H NMR (400 MHz, DMSO-d6) δ 7.10 (s, 2H) , 6.18 (s, 1H) , 4.58 (d, J = 5.2 Hz, 1H) , 4.14 (d, J = 11.9 Hz, 1H) , 4.07 –3.94 (m, 1H) , 3.88 –3.76 (m, 5H) , 3.71 (dd, J = 11.2, 2.7 Hz, 1H) , 3.62 –3.51 (m, 1H) , 3.24 –3.02 (m, 2H) , 2.29 (s, 3H) , 2.21 –2.07 (m, 2H) , 1.66 (d, J = 13.2 Hz, 2H) , 1.22 (t, J = 7.5 Hz, 3H) .

Example 17

Synthesis of (R) -dicyclopropyl ( (3- (3-methyl-1H-pyrazol-5-yl) -5- (3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) -l6-sulfanone

Step 1. sulfinyldicyclopropane

To a solution of sulfurous dichloride (0.511 mL, 7.044 mmol) in THF was added C₃H₅MgBr (0.5M in THF, 42.3 mL, 21.1 mmol) at 0℃. The reaction was stirred at 0℃ for 1 hr. LC-MS showed the reaction was complete. The reaction mixture was diluted with EA (20 mL*3) , washed with saturated NH₄Cl and brine, dried over anhydrous Na₂SO₄, filtered and concentrated. The residue was purified by flash column chromatography (Silica gel, 0～10%, MeOH in DCM) to give sulfinyldicyclopropane (400 mg, 43.61%) as a brown liquid. LC/MS (ESI) : m/z 130 (M+H) ⁺.

Step 2. N- (dicyclopropyl (oxo) -l6-sulfanylidene) -2, 2, 2-trifluoroacetamide

To a solution of sulfinyldicyclopropane (350 mg, 2.69 mmol) in DCM (20 mL) was added Rh (OAc) ₂ (24 mg, 0.054 mmol) , MgO (542 mg, 13.4 mmol) , Trifluoroacetamide (608 mg, 5.38 mmol) and PhI (OAc) ₂ (1.3 g, 4.032 mmol ) . The reaction was stirred at room temperature for 15 h under nitrogen atmosphere. LC-MS showed the reaction was complete. The reaction mixture is filtered and concentrated. The residue was purified by flash column chromatography (silica gel, 0～10%, MeOH in DCM) to afford N- (dicyclopropyl (oxo) -l6-sulfanylidene) -2, 2, 2-trifluoroacetamide (179 mg, 0.742 mmol, 27.61%) as a yellow solid. LC/MS (ESI) : m/z 241 (M+H) ⁺.

Step 3. dicyclopropyl (imino) -l6-sulfanone

To a solution of N- (dicyclopropyl (oxo) -l6-sulfanylidene) -2, 2, 2-trifluoroacetamide (174 mg, 0.721 mmol) in MeOH (2 mL) was added K₂CO₃ (179 mg, 1.29 mmol) . The mixture was stirred at room temperature for 2 h. LC-MS showed the reaction was complete. The reaction mixture is filtered and concentrated. The residue was purified by prep-HPLC (C₁₈, 10-95%, MeOH in H₂O with 0.1% HCOOH) to afford dicyclopropyl (imino) -l6-sulfanone (47 mg, 0.324 mmol, 44.87%) as a yellow liquid. LC/MS (ESI) : m/z 145 [M+H] ⁺.

Step 4. dicyclopropyl ( (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) -l6-sulfanone

To a solution of (3R) -4- {7-chloro-3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine (94 mg, 0.217 mmol) in dioxane (3 mL) was added dicyclopropyl (imino) -l6-sulfanone (47 mg, 0.325 mmol) , Pd₂ (dba) ₃ (21 mg, 0.023 mmol) , Xantphos (26 mg, 0.046 mmol) and Cs₂CO₃ (250 mg, 0.767 mmol) . The reaction was stirred at 100℃ overnight under nitrogen atmosphere. LC-MS showed the reaction was complete. The reaction mixture is filtered and concentrated in vacuo. The residue was purified by flash column chromatography (silica gel, 0～100%, EA in PE) to afford dicyclopropyl ( (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) -l6-sulfanone (84 mg, 0.155 mmol, 71.45%) as a yellow solid. LC/MS (ESI) : m/z 542 (M+H) ⁺.

Step 5. (R) -dicyclopropyl ( (3- (3-methyl-1H-pyrazol-5-yl) -5- (3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) -l6-sulfanone

To a solution of dicyclopropyl ( (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) -l6-sulfanone (84 mg, 0.155 mmol ) in DCM (1 mL) was added HCl (4M in dioxane, 2 mL) . The mixture was stirred at room temperature for 1 h. LC-MS showed the reaction was complete. The reaction mixture was concentrated under vacuo. The residue was purified by prep-HPLC (C₁₈, 10-95%, MeOH in H₂O with 0.1% HCOOH) to afford (R) -dicyclopropyl ( (3- (3-methyl-1H-pyrazol-5-yl) -5- (3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) -l6-sulfanone (30 mg, 42.3%) . LC/MS (ESI) : m/z 458 [M+H] ⁺. ¹H NMR (400 MHz, DMSO-d6) δ: 13.11 (s, 1H) , 7.09 (s, 1H) , 6.55 (s, 1H) , 4.28 (s, 1H) , 4.02 (d, J = 8.4 Hz, 1H) , 3.92 (s, 1H) , 3.79 (d, J = 11.1 Hz, 1H) , 3.70 (d, J = 8.7 Hz, 1H) , 3.55 (t, J = 10.3 Hz, 1H) , 3.17 (t, J = 11.0 Hz, 1H) , 3.06 –2.94 (m, 2H) , 2.31 (d, J = 17.9 Hz, 3H) , 1.39 –1.21 (m, 5H) , 1.19 (d, J = 6.5 Hz, 3H) , 1.18 –1.10 (m, 3H) .

Example 18

Step 1. (3R) -3-methyl-4- {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -7- [2- (oxan-2-yl) -2H-1, 2, 3-triazol-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} morpholine

A mixture of (3R) -4- {7-chloro-3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine (100 mg, 0.230 mmol) in Dioxane (5 mL) and H₂O (1 mL) was added 2- (oxan-2-yl) -4- (tetramethyl-1, 3, 2- dioxaborolan-2-yl) -2H-1, 2, 3-triazole (77 mg, 0.277 mmol) , K₂CO₃ (95 mg, 0.691 mmol) and tetrakis (triphenylphosphane) palladium (27 mg, 0.023 mmol) . The reaction was stirred at 100℃ for 15 h under N₂ atmosphere. LC-MS showed the reaction was complete. The reaction mixture was diluted with EtOAc (100 mL) , then washed with water and brine, dried over anhydrous Na₂SO₄, filtered and concentrated. The residue was purified by column chromatography on silica gel (PE: EA = 1: 1) to afford (3R) -3-methyl-4- {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -7- [2- (oxan-2-yl) -2H-1, 2, 3-triazol-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} morpholine (40 mg, 29.95%) as a yellow solid. LC/MS (ESI) : m/z 552.0 [M+H] ⁺.

Step 2. (3R) -3-methyl-4- [3- (3-methyl-1H-pyrazol-5-yl) -7- (2H-1, 2, 3-triazol-4-yl) - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] morpholine

To a solution of (3R) -3-methyl-4- {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -7- [2- (oxan-2-yl) -2H-1, 2, 3-triazol-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} morpholine (50 mg, 0.091 mmol) in DCM (3 mL) was added HCl (4M in dioxane, 1 mL) at room temperature. The mixture was stirred at rt for 1 hour. LC-MS showed the reaction was complete. The reaction mixture was concentrated under reduced pressure. The residue was purified by prep-HPLC (C₁₈, 10-95%, MeOH in H₂O with 0.1%HCOOH) to give (3R) -3-methyl-4- [3- (3-methyl-1H-pyrazol-5-yl) -7- (2H-1, 2, 3-triazol-4-yl) [1, 2] thiazolo [4, 5-b] pyridin-5-yl] morpholine (10 mg, 0.026 mmol, 28.8%) . LC/MS (ESI) : m/z 383.1 [M+H] ⁺. ¹H NMR (400 MHz, DMSO-d6) δ 9.01 (s, 1H) , 7.77 (d, J = 15.6 Hz, 1H) , 7.17 (d, J = 13.4 Hz, 1H) , 4.62 (d, J = 5.8 Hz, 1H) , 4.19 (d, J = 10.9 Hz, 1H) , 4.10 –4.05 (m, 1H) , 3.85 (s, 1H) , 3.75 (s, 1H) , 3.71 (s, 1H) , 3.30 (d, J = 3.6 Hz, 1H) , 2.32 (s, 3H) , 1.28 (t, J = 9.2 Hz, 3H) .

Example 19

Synthesis of (R) - (1- (3- (3-methyl-1H-pyrazol-5-yl) -5- (3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) -1H-1, 2, 4-triazol-3-yl) methanol

Step 1. 1- (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) -1H-1, 2, 4-triazole-3-carboxylic

To a mixture of (3R) -4- {7-chloro-3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine (100 mg, 0.230 mmol) and Cs₂CO₃ (224 mg, 0.690 mmol) in DMA (1 mL) was added methyl 1H-1, 2, 4-triazole-5-carboxylate (59 mg, 0.461 mmol) . The reaction was stirred at 140℃ for 2 h. LC-MS showed the reaction was completed. The reaction mixture was adjusted with HCl (1 M) to pH=7 and extracted with EA (20 mL*5) . The combined organic phase was washed with brine (20 mL) , dried over anhydrous Na₂SO₄, filtered and concentrated to dryness. The residue was purified by Prep-TLC (DCM: MeOH = 10: 1, V/V) to afford 1- (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) -1H-1, 2, 4-triazole-3-carboxylic acid (70 mg, 0.137 mmol, 59.50%) as a yellow solid. LC/MS (ESI) : m/z 511.6 (M+H) ⁺.

Step 2. (1- (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) -1H-1, 2, 4-triazol-3-yl) methanol

To a solution of 1- (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) -1H-1, 2, 4-triazole-3-carboxylic acid (60 mg, 0.118 mmol) in THF (2 mL) was added LiAlH₄ (10 mg, 0.264 mmol) at -78℃. The mixture was stirred at -78℃ for 1 h. LC-MS showed the reaction was completed. The reaction mixture was quenched with Na₂SO₄ ^.10H₂O, filtered and concentrated. The residue was purified by Prep-TLC (DCM: MeOH = 10: 1, V/V) to afford (1- (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) -1H-1, 2, 4-triazol-3-yl) methanol (18 mg, 0.036 mmol, 30.84%) as a yellow solid. LC/MS (ESI) : m/z 497.6 [M+H] ⁺.

Step 3. (R) - (1- (3- (3-methyl-1H-pyrazol-5-yl) -5- (3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) -1H-1, 2, 4-triazol-3-yl) methanol

To a solution of (1- (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) -1H-1, 2, 4-triazol-3-yl) methanol (16 mg, 0.032 mmol) in DCM (1 mL) was added HCl (4 M in dioxane, 1.5 mL) . The mixture was stirred at room temperature for 0.5 h. LC-MS showed the reaction was completed. The reaction mixture was concentrated. The residue was purified by Prep-HPLC (C₁₈, 10-95%, MeOH in H₂O with 0.1%HCOOH) to afford (R) - (1- (3- (3-methyl-1H-pyrazol-5-yl) -5- (3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) -1H-1, 2, 4-triazol-3-yl) methanol (5 mg, 0.012 mmol, 37.62%) . LC/MS (ESI) : m/z: 413 [M+H] ⁺. ¹H NMR (400 MHz, DMSO) δ 13.19 (s, 1H) , 9.81 (s, 1H) , 7.77 (s, 1H) , 7.12 (s, 1H) , 5.57 (s, 1H) , 4.61 (d, J = 12.4 Hz, 3H) , 4.19 (d, J = 12.7 Hz, 1H) , 4.08 (d, J = 8.5 Hz, 1H) , 3.86 (d, J = 11.2 Hz, 1H) , 3.75 (d, J = 8.9 Hz, 1H) , 3.60 (t, J = 10.5 Hz, 1H) , 3.29 –3.26 (m, 1H) , 2.31 (s, 3H) , 1.28 (d, J = 6.6 Hz, 3H) .

Example 20

Synthesis of 1- { [3- (3-methyl-1H-pyrazol-5-yl) -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl] imino} -1λ^6-thiolan-1-one

Step 1. 1- ( {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} imino) -1λ^6-thiolan-1-one

To a solution of (3R) -4- {7-chloro-3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine (50 mg, 0.115 mmol) in Dioxane (5 mL) was added 1-imino-1λ^6-thiolan-1-one (17 mg, 0.138 mmol) , Xantphos (27 mg, 0.046 mmol) , Cs₂CO₃ (113 mg, 0.346 mmol) and Pd₂ (dba) ₃ (21 mg, 0.023 mmol) . The reaction was stirred at 100℃ for 15 h under N₂ atmosphere. TLC showed the reaction was complete. The mixture was added H₂O (10 mL) and extracted with EtOAc (3*20 mL) . The organic layer was washed with brine, dried over Na₂SO₄, filtered and concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel (0-100%, EA in Pe) to give 1- ( {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} imino) -1λ^6-thiolan-1-one (40 mg, 64%) as a yellow oil. LC/MS (ESI) : m/z 517.2 [M+H] ⁺.

Step 2. 1- { [3- (3-methyl-1H-pyrazol-5-yl) -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl] imino} -1λ^6-thiolan-1-one

To a solution of 1- ( {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} imino) -1λ^6-thiolan-1-one (40 mg, 0.077 mmol) in DCM (4 mL) was added HCl (4M in dioxane, 1 mL) . The mixture was stirred at room temperature for 1 hour. TLC showed the reaction was complete. The reaction mixture was concentrated under reduced pressure. The residue was purified by Prep-HPLC (C₁₈, 10-95%, MeOH in H₂O with 0.1%HCOOH) to give 1- { [3- (3-methyl-1H-pyrazol-5-yl) -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl] imino} -1λ^6-thiolan-1-one (10 mg, 29.86%) . LC/MS (ESI) : m/z 433.1 [M+H] ⁺. ¹H NMR (400 MHz, DMSO-d6) δ 12.98 (NH, 1H) , 7.10 (s, 1H) , 6.44 (s, 1H) , 4.37 (d, J = 5.2 Hz, 1H) , 4.00 (dd, J = 20.5, 8.8 Hz, 2H) , 3.84 –3.68 (m, 2H) , 3.64 –3.50 (m, 5H) , 3.19 (td, J = 12.4, 3.2 Hz, 1H) , 2.58 –2.43 (m, 2H) , 2.20 –2.10 (m, 2H) , 1.21 (d, J = 6.6 Hz, 3H) , 1.21 (d, J = 6.6 Hz, 3H) .

Example 21

Synthesis of (R) -1- ( (3- (3-methyl-1H-pyrazol-5-yl) -5- (3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) hexahydro-1l6-thiopyran 1-oxide

Step 1. 1- ( (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) hexahydro-1l6-thiopyran 1-oxide

To a solution of (3R) -4- {7-chloro-3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine (100 mg, 0.230 mmol) in dioxane (3 mL) was added (dicyclopropylimino-λ^6-sulfanyl) one (47 mg, 0.325 mmol) , Pd₂ ₍dba) ₃ (21 mg, 0.023 mmol) , Xantphos (27 mg, 0.046 mmol) and Cs₂CO₃ (250 mg, 0.767 mmol) . The reaction was stirred at 100℃ overnight under nitrogen atmosphere. TLC showed the reaction was complete. The reaction mixture was filtered and concentrated in vacuo. The residue was purified by flash column chromatography (Silica gel, 0～100%ethyl acetate in petroleum ether) to afford 1- ( (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) hexahydro-1l6-thiopyran 1-oxide (80 mg, 65.41%) as a yellow solid. LC/MS (ESI) : m/z 531.2 [M+H] ⁺.

Step 2. (R) -1- ( (3- (3-methyl-1H-pyrazol-5-yl) -5- (3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) hexahydro-1l6-thiopyran 1-oxide

To a solution of 1- ( (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) hexahydro-1l6-thiopyran 1-oxide (80 mg, 0.151 mmol) in DCM (1 mL) was added HCl/dioxane (2 mL) . The mixture was stirred at room temperature for 1 h. LC-MS showed the reaction was complete. The reaction mixture was concentrated under vacuo. The residue was purified by prep-HPLC (C₁₈, 10-95%, MeOH in H₂O with 0.1%HCOOH) to afford 1- { [3- (3-methyl-1H-pyrazol-5-yl) -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl] imino} -1λ^6-thian-1-one (30 mg, 44.56%) . LC/MS (ESI) : m/z 447 [M+H] ⁺. ¹H NMR (400 MHz, DMSO-d6) δ 7.14 (s, 1H) , 6.60 (s, 1H) , 4.42 (s, 1H) , 4.07 (d, J = 8.3 Hz, 1H) , 4.01 (d, J = 11.8 Hz, 1H) , 3.86 (d, J =11.0 Hz, 1H) , 3.71 (dd, J = 38.8, 16.0 Hz, 5H) , 3.25 (s, 2H) , 2.36 (s, 3H) , 2.01 (d, J =38.4 Hz, 4H) , 1.72-1.68 (m, 2H) , 1.28 (m, 3H) .

Example 22

Synthesis of (R) -4- (7- (1- (difluoromethyl) cyclopropyl) -3- (3-methyl-1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine

Step 1. (3R) -4- (7- (1- (difluoromethyl) cyclopropyl) -3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine

To a mixture of (R) -4- (3-chloro-7- (1- (difluoromethyl) cyclopropyl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine (50 mg, 0.139 mmol) , (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) boronic acid (58 mg, 0.278 mmol) and K₂CO₃ (96 mg, 0.695 mmol) in dioxane (5 mL) and water (1 mL) was added Pd (PPh₃) ₄ (16 mg, 0.014 mmol) . The mixture was degassed and charged with N₂ for several times and then stirred at 80℃ for 16 h under N₂. LC-MS showed the reaction was completed. The reaction mixture was poured into H₂O (30 mL) and extracted with EA (30 mL *3) . The combined organic phase was washed with brine (30 mL) , dried over anhydrous Na₂SO₄, filtered and concentrated to dryness. The residue was purified by column chromatography (silica gel, 0-100%, EA in PE) to give (3R) -4- (7- (1- (difluoromethyl) cyclopropyl) -3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine (40 mg, 0.082 mmol, 58.80%) as a yellow solid. LC/MS (ESI) : m/z 490 [M+H] ⁺.

Step 2. (R) -4- (7- (1- (difluoromethyl) cyclopropyl) -3- (3-methyl-1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine

To a solution of (3R) -4- (7- (1- (difluoromethyl) cyclopropyl) -3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine (40 mg, 0.082 mmol) in MeOH (2 mL) was added TsOH (14 mg, 0.082 mmol) . The mixture was stirred at room temperature for 1 h. LC-MS showed the reaction was completed. The reaction mixture was poured into saturated NaHCO₃ and extracted with EA (30 mL *3) . The combined organic phase was washed with brine, dried over anhydrous Na₂SO₄, filtered and concentrated to dryness. The residue was purified by Prep-HPLC (C₁₈, 10-95%, MeOH in H₂O with 0.1%HCOOH) to afford (R) -4- (7- (1- (difluoromethyl) cyclopropyl) -3- (3-methyl-1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine (12 mg, 0.029 mmol, 72.07%) . LC/MS (ESI) : m/z: 406 [M+H] ⁺. ¹H NMR (400 MHz, DMSO) δ 7.23 (s, 1H) , 7.11 (s, 1H) , 5.98 (s, 1H) , 4.51 (s, 1H) , 4.17 –3.96 (m, 2H) , 3.81 (d, J = 11.6 Hz, 1H) , 3.71 (d, J = 8.9 Hz, 1H) , 3.56 (s, 1H) , 3.17 (d, J = 5.3 Hz, 1H) , 2.31 (d, J = 18.4 Hz, 3H) , 1.31 (s, 2H) , 1.23 (d, J = 6.9 Hz, 5H) .

Example 23

Synthesis of (R) -1- ( (3- (3-methyl-1H-pyrazol-5-yl) -5- (3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) -1l6-thietane 1-oxide

Step 1. 1- ( (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) -1l6-thietane 1-oxide

To a solution of (3R) -4- {7-chloro-3- [1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine (100 mg, 0.238 mmol) in dioxane (4 mL) was added 1-imino-1λ^6-thietan-1-one (30 mg, 0.286 mmol) , Pd₂ (dba) ₃ (22 mg, 0.024 mmol) , Xantphos (28 mg, 0.048 mmol) and C_S2CO₃ (233 mg, 0.714 mmol) . The reaction was stirred at 100℃ overnight under nitrogen atmosphere. LC-MS showed the reaction was complete. The reaction was filtered and concentrated in vacuo. The residue was purified by flash column chromatography (Silica gel, 0～100%ethyl acetate in petroleum ether) to afford 1- ( (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) -1l6-thietane 1-oxide (80 mg, 0.160 mmol, 67.13%) as a yellow solid. LC/MS (ESI) : m/z 501 (M+H) ⁺.

Step 2. (R) -1- ( (3- (3-methyl-1H-pyrazol-5-yl) -5- (3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) -1l6-thietane 1-oxide

To a solution of 1- ( {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} imino) -1λ^6-thietan-1-one (20 mg, 0.040 mmol) in MeOH (2 mL) was added TsOH (2 mg, 0.040 mmol) . The mixture was stirred at room temperature for 1 h. LC-MS showed the reaction was complete. The reaction mixture was concentrated under vacuo. The residue was purified by prep-HPLC (C₁₈, 10-95%, MeOH in H₂O with 0.1%HCOOH) to afford (R) -1- ( (3- (3-methyl-1H-pyrazol-5-yl) -5- (3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) -1l6-thietane 1-oxide (12 mg, 0.029 mmol, 72.07%) . LC/MS (ESI) : m/z 418 [M+H] ⁺. ¹H NMR (400 MHz, DMSO-d6) δ 7.10 (s, 1H) , 6.38 (s, 1H) , 4.50 (dd, J = 15.3, 6.9 Hz, 2H) , 4.41 (dd, J = 9.8, 4.4 Hz, 2H) , 4.36 (s, 1H) , 3.82 –3.67 (m, 2H) , 3.55 (dd, J = 11.7, 8.8 Hz, 2H) , 3.24 –3.15 (m, 2H) , 2.40 (dd, J = 10.6, 4.9 Hz, 2H) , 2.29 (s, 3H) , 1.20 (d, J = 6.7 Hz, 3H) .

Example 24

Synthesis of (ethyl (methyl) ( (3- (3-methyl-1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) -l6-sulfanone

Step 1. ethyl (methyl) ( (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) -l6-sulfanone

To a solution of (3R) -4- {7-chloro-3- [1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine (150 mg, 0.357 mmol) in dioxane (4 mL) was added ethyl (imino) methyl-λ^6-sulfanone (57 mg, 0.536 mmol) , Pd₂(dba) ₃ (33 mg, 0.036 mmol) , Xantphos (41 mg, 0.071 mmol) and Cs₂CO₃ (349 mg, 1.07 mmol) . The reaction was stirred at 100℃ overnight under nitrogen atmosphere. LC-MS showed the reaction was complete. The reaction was filtered and concentrated in vacuo. The residue was purified by flash column chromatography (Silica gel, 0～100%ethyl acetate in petroleum ether) to afford ethyl (methyl) ( (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) -l6-sulfanone (120 mg, 66.83%) as a yellow solid. LC/MS (ESI) m/z: 503 (M+H) ⁺.

Step 2. ethyl (methyl) ( (3- (3-methyl-1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) -l6-sulfanone

To a solution of ethyl (methyl) ( (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) -l6-sulfanone (100 mg, 0.198 mmol) in DCM (1 mL) was added HCl/dioxane (2 mL) . The mixture was stirred at room temperature for 1 h. LC-MS showed the reaction was complete. The reaction mixture was concentrated under vacuo. The residue was purified by SFC to afford 24a (16 mg, 19.16%) and 24b (12 mg, 14.37%) . LC/MS (ESI) m/z: 420 [M+H] ⁺.

24a:

¹H NMR (400 MHz, DMSO-d6) δ 7.08 (s, 1H) , 6.56 (s, 1H) , 4.36 (d, J = 6.8 Hz, 1H) , 4.07 –3.90 (m, 2H) , 3.80 (d, J = 11.3 Hz, 1H) , 3.71 (dd, J = 11.3, 2.7 Hz, 1H) , 3.62 (d, J = 7.3 Hz, 2H) , 3.39 (s, 3H) , 3.19 (td, J = 12.5, 3.6 Hz, 2H) , 2.29 (s, 3H) , 1.35 (t, J = 7.3 Hz, 3H) , 1.20 (d, J = 6.7 Hz, 3H) .

24b:

¹H NMR (400 MHz, DMSO-d6) δ 7.08 (s, 1H) , 6.56 (s, 1H) , 4.35 (d, J = 6.5 Hz, 1H) , 4.08 –3.91 (m, 2H) , 3.80 (d, J = 11.3 Hz, 1H) , 3.71 (dd, J = 11.3, 2.7 Hz, 1H) , 3.62 (d, J = 7.4 Hz, 2H) , 3.40 (s, 3H) , 3.19 (td, J = 12.6, 3.6 Hz, 2H) , 2.29 (s, 3H) , 1.35 (t, J = 7.3 Hz, 3H) , 1.20 (d, J = 6.6 Hz, 3H) .

Example 25

Synthesis of (R) -4- (7- (1- (fluoromethyl) cyclopropyl) -3- (3-methyl-1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine

Step 1. (3R) -4- (7- (1- (fluoromethyl) cyclopropyl) -3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine

To a mixture of (R) -4- (3-chloro-7- (1- (fluoromethyl) cyclopropyl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine (30 mg, 0.088 mmol) , (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) boronic acid (37 mg, 0.176 mmol) and K₂CO₃ (61 mg, 0.439 mmol) in dioxane (5 mL) and water (1 mL) was added Pd (PPh₃) ₄ (10 mg, 0.009 mmol) . The mixture was degassed and charged with N₂ for several times and then stirred at 80℃ for 16 h under N₂. LC-MS showed the reaction was completed. The reaction mixture was poured into H₂O (30 mL) and extracted with EA (30 mL *3) . The combined organic phase was washed with brine (30 mL) , dried over anhydrous Na₂SO₄, filtered and concentrated to dryness. The residue was purified by Prep-TLC (PE: EA=1: 1) to give (3R) -4- (7- (1- (fluoromethyl) cyclopropyl) -3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine (20 mg, 0.042 mmol, 48.32%) as a yellow solid. LC/MS (ESI) : m/z 472 [M+H] ⁺.

Step 2. (R) -4- (7- (1- (fluoromethyl) cyclopropyl) -3- (3-methyl-1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine

To a solution of (3R) -4- (7- (1- (fluoromethyl) cyclopropyl) -3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine (20 mg, 0.042 mmol) in MeOH (2 mL) was added TsOH (7 mg, 0.042 mmol) . The mixture was stirred at room temperature for 1 h. LC-MS showed the reaction was completed. The reaction mixture was poured into saturated NaHCO₃ and extracted with EA (30 mL *3) . The combined organic phase was washed with brine (30 mL) , dried over anhydrous Na₂SO₄, filtered and concentrated to dryness. The residue was purified by Prep-HPLC (C₁₈, 10-95%, MeOH in H₂O with 0.1% HCOOH) to afford (R) -4- (7- (1- (fluoromethyl) cyclopropyl) -3- (3-methyl-1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine (2 mg, 0.005 mmol, 12.17%) . LC/MS (ESI) : m/z 387 [M+H] ⁺. ¹H NMR (400 MHz, DMSO) δ 13.91 –12.11 (m, 1H) , 7.23 (s, 1H) , 7.12 (s, 1H) , 4.63 (s, 1H) , 4.17 (d, J = 13.0 Hz, 1H) , 4.04 (d, J = 10.2 Hz, 1H) , 3.81 (s, 1H) , 3.73 (d, J = 10.1 Hz, 1H) , 3.58 (s, 1H) , 3.24 (s, 1H) , 2.78 –2.63 (m, 4H) , 2.31 (s, 3H) , 2.12 (s, 1H) , 1.88 (d, J = 9.0 Hz, 1H) , 1.24 (d, J = 6.1 Hz, 3H) .

Example 26

Synthesis of cyclopropyl (methyl) ( (3- (3-methyl-1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) -l6-sulfanone

Step 1. (methylsulfinyl) cyclopropane

To a solution of 1-bromo-4-methanesulfinylbenzene (1.8 mL, 13.7 mmol) in THF (30 mL) was added C₃H₅MgBr (0.5M in THF, 41 mL, 20.5 mmol) at 0℃ slowly. The mixture was stirred at 0℃ for 1.5h. LC-MS showed the reaction was complete. The mixture was poured into saturated NH₄Cl and extracted with EA (20 mLx3) . The combined organic layer was washed with brine, dried over anhydrous Na₂SO₄, filtered and concentrated. The residue was purified by flash chromatography (silica gel, 0 ～ 50 %, ethyl acetate in petroleum ether) to afford (methylsulfinyl) cyclopropane (900 mg, 8.64 mmol, 63.10%) as a yellow oil. LC/MS (ESI) : m/z 105 [M+H] ⁺.

Step 2. cyclopropyl (imino) methyl-λ^6-sulfanone

To a solution of (methylsulfinyl) cyclopropane (1 g, 9.600 mmol) ) , MgO (1.55 g, 38.4 mmol) , PhI (OAc) ₂ (4.64 g, 14.4 mmol) and Rh (OAc) ₂ (4.23 g, 9.60 mmol) in DCM (30 mL) was added NH₂COONH₄ (1.12 g, 14.4 mmol) . The reaction mixture was stirred at 25℃ for 16h. LC-MS showed the reaction was complete. The mixture was filtered and concentrated. The residue was purified by flash chromatography (silica gel, 0 ～ 10 %, MeOH in DCM) to afford cyclopropyl (imino) methyl-λ^6-sulfanone (300 mg, 2.52 mmol, 26.22%) as a colorless oil. LC/MS (ESI) : m/z 120 [M+H] ⁺.

Step 3. cyclopropyl (methyl) ( (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) -l6-sulfanone

To a solution of (3R) -4- {7-chloro-3- [1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine (150 mg, 0.357 mmol) in dioxane (4 mL) was added cyclopropyl (imino) methyl-λ^6-sulfanone (64 mg, 0.535 mmol) , Pd₂ (dba) ₃ (33 mg, 0.036 mmol) , Xantphos (41 mg, 0.071 mmol) and C_S2CO₃ (349 mg, 1.07 mmol) . The reaction was stirred at 100℃ overnight under nitrogen atmosphere. LC-MS showed the reaction was complete. The reaction was filtered and concentrated in vacuo. The residue was purified by flash chromatography (silica gel, 0 ～ 100 %, ethyl acetate in petroleum ether) to afford cyclopropyl (methyl) ( (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) -l6-sulfanone (60 mg, 0.117 mmol, 32.65%) as a yellow solid. LC/MS (ESI) : m/z 515 (M+H) ^+.

Step 4. cyclopropyl (methyl) ( (3- (3-methyl-1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) -l6-sulfanone

To a solution of cyclopropyl (methyl) ( (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7- yl) imino) -l6-sulfanone (80 mg, 0.155 mmol) in DCM (1 mL) was added HCl/dioxane (2 mL) . The mixture was stirred at room temperature for 1 h. LC-MS showed the reaction was complete. The reaction mixture was concentrated under vacuo. The residue was purified by SFC to afford 26a (10 mg, 0.023 mmol, 14.93%) and 26b (5 mg, 0.012 mmol, 7.47%) . LC/MS (ESI) : m/z 420 [M+H] ⁺.

26a:

¹HNMR (400 MHz, DMSO-d6) δ 13.10 (s, 1H) , 7.09 (s, 1H) , 6.55 (s, 1H) , 4.34 (d, J = 5.4 Hz, 1H) , 4.01 (d, J = 8.3 Hz, 1H) , 3.91 (s, 1H) , 3.80 (d, J = 11.2 Hz, 1H) , 3.70 (dd, J = 11.3, 2.7 Hz, 1H) , 3.55 (d, J = 2.8 Hz, 1H) , 3.45 (s, 3H) , 3.18 (dd, J = 12.5, 9.4 Hz, 1H) , 3.08 –3.00 (m, 1H) , 2.29 (s, 3H) , 1.40 –1.31 (m, 1H) , 1.24 (s, 2H) , 1.21 –1.08 (m, 5H) .

26b:

¹H NMR (400 MHz, DMSO-6d) δ 13.08 (s, 1H) , 7.08 (s, 1H) , 6.55 (s, 1H) , 4.28 (d, J = 6.5 Hz, 1H) , 4.00 (t, J = 13.1 Hz, 2H) , 3.79 (d, J = 11.4 Hz, 1H) , 3.75 –3.54 (m, 2H) , 3.55 (t, J = 10.3 Hz, 1H) , 3.46 (s, 3H) , 3.18 (dd, J = 12.6, 9.1 Hz, 1H) , 3.02 (s, 1H) , 2.29 (s, 3H) , 1.42 –1.33 (m, 1H) , 1.24 (s, 2H) , 1.20 (d, J = 6.7 Hz, 3H) , 1.15 (s, 1H) .

Example 27

Synthesis of (R) -4- (7- (1- (difluoromethyl) cyclopropyl) -3- (1H-pyrazol-5- yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine

Step 1. (R) -4- (3-chloro-7- (1- (difluoromethyl) cyclopropyl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine

To a solution of (R) -1- (3-chloro-5- (3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) cyclopropane-1-carbaldehyde (100 mg, 0.296 mmol) in DCM (5 mL) was added DAST (143 mg, 0.888 mmol) at 0℃. The mixture was stirred at 0℃ for 1 h. LC-MS showed the reaction was completed. The mixture was poured into saturated NaHCO₃ (30 mL) and extracted with DCM (30 mL*3) . The combined organic phase was washed with brine (30 mL) , dried over anhydrous Na₂SO₄, filtered and concentrated in vacuo. The residue was purified by column chromatography (silica gel, 0-100%, EA in PE) to give (R) -4- (3-chloro-7- (1- (difluoromethyl) cyclopropyl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine (50 mg, 0.139 mmol, 46.94%) as a white solid. LC/MS (ESI) : m/z 360.8 [M+H] ⁺.

Step 2. (3R) -4- (7- (1- (difluoromethyl) cyclopropyl) -3- (1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine

To a mixture of (R) -4- (3-chloro-7- (1- (difluoromethyl) cyclopropyl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine (50 mg, 0.139 mmol) , 1- (tetrahydro-2H-pyran-2-yl) -5- (4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole (58 mg, 0.208 mmol) , dipotassium carbonate (58 mg, 0.417 mmol) in dioxane (5 mL) and water (1 mL) was added Pd (PPh₃) ₄ (35 mg, 0.030 mmol) . The mixture was degassed and charged with N₂ for several times and then stirred at 80℃ for 16 h under N₂. The mixture was poured into H₂O (20 mL) and extracted with EA (20 mL *3) . The combined organic phase was washed with brine, dried over anhydrous Na₂SO₄, filtered and concentrated in vacuo. The residue was purified by Prep-TLC (PE: EA=1: 1) to give (3R) -4- (7- (1- (difluoromethyl) cyclopropyl) -3- (1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine (15 mg, 0.032 mmol, 56.75%) as a yellow solid. LC/MS (ESI) : m/z 476.6 [M+H] ⁺.

Step 3. (R) -4- (7- (1- (difluoromethyl) cyclopropyl) -3- (1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine

To a solution of (3R) -4- (7- (1- (difluoromethyl) cyclopropyl) -3- (1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine (15 mg, 0.032 mmol) in MeOH (1 mL) was added TsOH (5.5 mg, 0.032 mmol) . The mixture was stirred at room temperature for 1 h. LC-MS showed the reaction was completed. The mixture was poured into saturated NaHCO₃ and extracted with EA (20 mL *3) . The combined organic phase was washed with brine (20 mL) , dried over anhydrous Na₂SO₄, filtered and concentrated in vacuo. The residue was purified by Prep-HPLC (C₁₈, 10-95%, MeOH in H₂O with 0.1%HCOOH) to afford (R) -4- (7- (1-(difluoromethyl) cyclopropyl) -3- (1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine (2 mg, 0.005 mmol, 12.17%) . LC/MS (ESI) : m/z 392 [M+H] ⁺. ¹H NMR (400 MHz, DMSO) δ 13.65 (s, 1H) , 7.74 (s, 1H) , 7.38 (d, J = 1.9 Hz, 1H) , 7.23 (s, 1H) , 5.99 (t, J = 55.6 Hz, 1H) , 4.53 (d, J = 4.7 Hz, 1H) , 4.22 –3.99 (m, 2H) , 3.81 (d, J = 11.3 Hz, 1H) , 3.71 (dd, J = 11.4, 2.8 Hz, 1H) , 3.59 –3.50 (m, 1H) , 3.23 (dd, J = 12.6, 3.7 Hz, 1H) , 1.31 (s, 2H) , 1.23 (d, J = 6.7 Hz, 5H) .

Example 28

Synthesis of dimethyl ( {5- [ (3R) -3-methylmorpholin-4-yl] -3- (1H-pyrazol-5-yl) - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} imino) -λ^6-sulfanone

Step 1. dimethyl ( {5- [ (3R) -3-methylmorpholin-4-yl] -3- [1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} imino) -λ^6-sulfanone

To a solution of (3R) -4- {7-chloro-3- [1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine (100 mg, 0.238 mmol) in dioxane (4 mL) was added iminodimethyl-λ^6-sulfanone (27 mg, 0.286 mmol) , Pd₂ (dba) ₃ (22 mg, 0.024 mmol) and Xantphos (28 mg, 0.048 mmol) , Cs₂CO₃ (233 mg, 0.714 mmol) . The reaction was stirred at 100℃ overnight under nitrogen atmosphere. LC-MS showed the reaction was complete. The reaction was diluted with EA (20 mL) and water (20 mL) . The organic layer was separated, washed with brine, dried over anhydrous Na₂SO₄, filtered and concentrated in vacuo. The residue was purified by column chromatography on silica gel (PE: EA = 2: 1, V/V) to afford dimethyl ( {5- [ (3R) -3-methylmorpholin-4-yl] -3- [1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} imino) -λ^6-sulfanone (72 mg, 63.4%) as a yellow solid. LC/MS (ESI) m/z: 478 (M+H) ⁺.

Step 2. dimethyl ( {5- [ (3R) -3-methylmorpholin-4-yl] -3- (1H-pyrazol-5-yl) - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} imino) -λ^6-sulfanone

To a solution of dimethyl ( {5- [ (3R) -3-methylmorpholin-4-yl] -3- [1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} imino) -λ^6-sulfanone (72 mg, 0.151 mmol) in DCM (1 mL) was added HCl (4M in dioxane, 2 mL) . The mixture was stirred at room temperature for 1 h. LC-MS showed the reaction was complete. The reaction mixture was concentrated under vacuo. The residue was purified by prep-HPLC (C₁₈, 10-95%, MeOH in H₂O with 0.1%HCOOH) to afford dimethyl ( {5- [ (3R) -3-methylmorpholin-4-yl] -3- (1H-pyrazol-5-yl) - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} imino) -λ^6-sulfanone (40 mg, 67.5%) . LC/MS (ESI) m/z: 393.1 (M+H) ⁺. ¹H NMR (400 MHz, DMSO-d6) δ 13.31 (s, 1H) , 7.71 (s, 1H) , 7.37 (d, J = 1.8 Hz, 1H) , 6.55 (s, 1H) , 4.39 (d, J = 6.7 Hz, 1H) , 4.03 –3.94 (m, 2H) , 3.80 (d, J = 11.2 Hz, 1H) , 3.70 (dd, J = 11.4, 2.8 Hz, 1H) , 3.55 (dd, J = 11.7, 8.9 Hz, 1H) , 3.48 (d, J = 1.2 Hz, 6H) , 3.23 –3.16 (m, 1H) , 1.21 (d, J = 6.6 Hz, 3H) .

Example 29

Synthesis of 2- {5- [ (3R) -3-methylmorpholin-4-yl] -3- (1H-pyrazol-5-yl) - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} propan-2-ol

Step 1. methyl 5- [ (3R) -3-methylmorpholin-4-yl] -3- [1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridine-7-carboxylate

To a solution of (3R) -4- {7-chloro-3- [1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine (200 mg, 0.476 mmol) in MeOH (3 mL) was added Pd (dppf) Cl₂ (70 mg, 0.095 mmol) , TEA (0.33 mL, 2.38 mmol) . The reaction was stirred at 65 ℃ overnight under CO atmosphere. LC-MS showed the reaction was complete. The reaction mixture was filtered and concentrated in vacuo. The residue was purified by flash column chromatography (Silica gel, 0～100%ethyl acetate in petroleum ether) to give methyl 5- [ (3R) -3-methylmorpholin-4-yl] -3- [1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridine-7-carboxylate (75 mg, 35.51%) as a yellow solid. LC/MS (ESI) : m/z 443 (M+H) ⁺.

Step 2. 2- {5- [ (3R) -3-methylmorpholin-4-yl] -3- [1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} propan-2-ol

A mixture of methyl 5- [ (3R) -3-methylmorpholin-4-yl] -3- [1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridine-7-carboxylate (75 mg, 0.169 mmol) in THF (2 mL) was added Methylmagnesiumbromid (0.34 mL, 0.845 mmol) . The reaction was stirred at 0℃ for 2 h. LC-MS showed the reaction was completed. 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 flash column chromatography (Silica gel, 0～100%ethyl acetate in petroleum ether) to give 2- {5- [ (3R) -3-methylmorpholin-4-yl] -3- [1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} propan-2-ol (75 mg, 0.169 mmol, 99.9%) as a white solid. LC/MS (ESI) : m/z 444 [M+H] ⁺.

Step 3. 2- {5- [ (3R) -3-methylmorpholin-4-yl] -3- (1H-pyrazol-5-yl) - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} propan-2-ol

To a solution of 2- {5- [ (3R) -3-methylmorpholin-4-yl] -3- [1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} propan-2-ol (75 mg, 0.169 mmol) in DCM (2 mL) was added HCl/dioxane (2 mL) . The mixture was stirred at room temperature for1 hour. LC-MS showed the reaction was complete. The reaction was concentrated in vacuo to give crude product, which was purified by prep-HPLC (C₁₈, 10-95%, MeCN in H₂O with 0.1%CHOOH) to afford the 2- {5- [ (3R) -3-methylmorpholin-4-yl] -3- (1H-pyrazol-5-yl) - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} propan-2-ol (14 mg, 0.039 mmol, 23.0%) . LC/MS (ESI) : m/z 361 (M+H) ⁺. ¹H NMR (400 MHz, DMSO-d6) δ 7.70 (s, 1H) , 7.36 (s, 1H) , 7.02 (s, 1H) , 6.10 (s, 1H) , 4.55 (s, 1H) , 4.07-4.03 (m, 2H) , 3.78-3.74 (m, 2H) , 3.48 (s, 1H) , 3.24 (d, J = 13.1 Hz, 1H) , 1.56 (s, 6H) , 1.20 (d, J = 6.0 Hz, 3H) .

Example 30

Synthesis of 4- {5- [ (3R) -3-methylmorpholin-4-yl] -3- [1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} oxan-4-ol

Step 1. 4- [7- (3, 6-dihydro-2H-pyran-4-yl) -3- [1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] -3-methylmorpholine

To a mixture of 4- {7-chloro-3- [1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine (100 mg, 0.238 mmol) , 2- (3, 6-dihydro-2H-pyran-4-yl) -4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolane (75 mg, 0.357 mmol) and K₂CO₃ (99 mg, 0.714 mmol) in Dioxane (2 mL) and H₂O (0.2 mL) was added Pd (PPh₃) ₄ (28 mg, 0.024 mmol) . The mixture was stirred at 100℃ for 16 h under N₂ atmosphere. LC-MS showed the reaction was complete. The reaction mixture was diluted with H₂O (20 mL) and extracted with EA (20 mL*3) . The combined organic layer was washed with brine, dried over anhydrous Na₂SO₄, filtered and concentrated. The residue was purified by column chromatography on silica gel (DCM: MeOH = 20: 1) to give the desired product 4- [7- (3, 6-dihydro-2H-pyran-4-yl) -3- [1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] -3-methylmorpholine (65 mg, 58.4%) as a brown solid. LC/MS (ESI) : m/z 468.1 [M+H] ⁺.

Step 2. 4- {5- [ (3R) -3-methylmorpholin-4-yl] -3- (1H-pyrazol-5-yl) - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} oxan-4-ol

A mixture of 4- [7- (3, 6-dihydro-2H-pyran-4-yl) -3- [1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] morpholine (60 mg, 0.132 mmol) , (4Z) -5- { [bis ( { [ (3Z) -2, 2, 6, 6-tetramethyl-5-oxohept-3-en-3-yl] oxy} ) ruthenio] oxy} -2, 6, 6-trimethylhept-4-en-3-one (8.4 mg, 0.013 mmol) and phenylsilane (0.033 mL, 0.265 mmol) in IPA (1 mL) and H₂O (0.1 mL) was stirred at rt for 2 h under O₂ atmosphere. LC-MS showed the reaction was complete. The reaction mixture was diluted with H₂O (20 mL) and extracted with EA (20 mL*3) . The combined organic layer was washed with brine, dried over anhydrous Na₂SO₄, filtered and concentrated. The residue was purified by column chromatography on silica gel (DCM: MeOH = 20: 1) to give 4- [5- (morpholin-4-yl) -3- [1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl] oxan-4-ol (20 mg, 0.042 mmol, 32.1%) as a brown solid. LC/MS (ESI) : m/z 472.1 [M+H] ^+.

Step 3. 4- {5- [ (3R) -3-methylmorpholin-4-yl] -3- (1H-pyrazol-5-yl) - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} oxan-4-ol

To a solution of 4- {5- [ (3R) -3-methylmorpholin-4-yl] -3- [1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} oxan-4-ol (20 mg, 0.04 mmol) in DCM (4 mL) was added HCl/Dioxane (4M, 1 mL) . The mixture was stirred at rt for 1 hour. TLC showed the reaction was complete. The reaction mixture was concentrated under reduced pressure. The residue was purified by prep-HPLC (C₁₈, 10-95%, MeOH in H₂O with 0.1%HCOOH) to give 4- {5- [ (3R) -3-methylmorpholin-4-yl] -3- (1H-pyrazol- 5-yl) - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} oxan-4-ol (10 mg, 0.025 mmol, 20.16%) . LC/MS (ESI) : m/z 402.1 [M+H] ⁺. ¹H NMR (400 MHz, DMSO-d6) δ 13.57 (s, 1H) , 7.68 (s, 1H) , 7.37 (d, J = 1.6 Hz, 1H) , 7.11 (s, 1H) , 6.22 (s, 1H) , 4.59 (d, J = 6.0 Hz, 1H) , 4.23 –4.04 (m, 1H) , 4.00-3.96 (m, 1H) , 3.90 –3.76 (m, 5H) , 3.71 (dd, J = 11.4, 2.7 Hz, 1H) , 3.54 (dt, J = 22.6, 14.5 Hz, 2H) , 3.27 –3.11 (m, 2H) , 2.29 –2.08 (m, 2H) , 1.66 (d, J = 13.4 Hz, 2H) , 1.35 –1.08 (m, 4H) .

Example 31

Synthesis of (R) -4- (7- (1- (fluoromethyl) cyclopropyl) -3- (1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine

Step 1. (R) -1- (3-chloro-5- (3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) cyclopropane-1-carbaldehyde

To a solution of (R) -1- (3-chloro-5- (3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) cyclopropane-1-carbonitrile (800 mg, 2.39 mmol) in Toluene (15 mL) was added DIBAL-H (1.0M in Toluene, 7.2 mL, 7.17 mmol) at -78℃. The mixture was stirred at -78℃ for 1 h. LC-MS showed the reaction was completed. The mixture was warmed to room temperature, poured into H₂SO₄ (1 M in water, 30 mL) and extracted with EA (50 mL *3) . The combined organic phase was washed with brine (30 mL) , dried over anhydrous Na₂SO₄, filtered and concentrated in vacuo. The residue was purified by column chromatography (silica gel, 0-100%, EA in PE) to give (R) -1- (3-chloro-5- (3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) cyclopropane-1-carbaldehyde (400 mg, 1.18 mmol, 49.56%) as a yellow oil. LC/MS (ESI) : m/z 338.8 [M+H] ⁺.

Step 2. (R) - (1- (3-chloro-5- (3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) cyclopropyl) methanol

To a solution of (R) -1- (3-chloro-5- (3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) cyclopropane-1-carbaldehyde (240 mg, 0.859 mmol) in MeOH (5 mL) was added NaBH₄ (90 mg, 2.37 mmol) . The mixture was stirred at 25℃ for 1h. LC-MS showed the reaction was complete. The mixture was poured into H₂O and extracted with DCM (30 mL*3) . The combined organic phase was washed with brine, dried over anhydrous Na₂SO₄, filtered and concentrated in vacuo. The residue was purified by column chromatography (silica gel, 0-100%, EA in PE) to give (R) - (1- (3-chloro-5- (3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) cyclopropyl) methanol (200 mg, 0.592 mmol, 50.00%) as a white solid. LC/MS (ESI) : m/z 340.8 [M+H] ⁺.

Step 3. (R) -4- (3-chloro-7- (1- (fluoromethyl) cyclopropyl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine

To a solution of (R) - (1- (3-chloro-5- (3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) cyclopropyl) methanol (100 mg, 0.294 mmol) in DCM (5 mL) was added DAST (285 mg, 1.77 mmol) at 0℃. The mixture was stirred at 0℃ for 1 h. LC-MS showed the reaction was completed. The mixture was poured into saturated NaHCO₃ and extracted with DCM (30 mL *3) . The combined organic phase was washed with brine (30 mL) , dried over anhydrous Na₂SO₄, filtered and concentrated in vacuo. The residue was purified by column chromatography (silica gel, 0-100%, EA in PE) to give (R) -4- (3-chloro-7- (1- (fluoromethyl) cyclopropyl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine (50 mg, 0.146 mmol, 49.71%) as a white solid. LC/MS (ESI) : m/z 342 [M+H] ⁺.

Step 4. (3R) -4- (7- (1- (fluoromethyl) cyclopropyl) -3- (1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine

To a mixture of (R) -4- (3-chloro-7- (1- (fluoromethyl) cyclopropyl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine (20 mg, 0.059 mmol) ) , 3-methyl-1- (oxan-2-yl) -5- (tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole (81 mg, 0.278 mmol) and K₂CO₃ (40.43 mg, 0.293 mmol) in dioxane (5 mL) and water (1 mL) was added Pd (PPh₃) ₄ (6.76 mg, 0.006 mmol) . The mixture was stirred at 80℃ for 16 h. The mixture was poured into H₂O (20 mL) and extracted with EA (20 mL *3) . The combined organic phase was washed with brine (20 mL) , dried over anhydrous Na₂SO₄, filtered and concentrated in vacuo. The residue was purified by Prep-TLC (PE: EA=1: 1) to give (3R) -4- (7- (1- (fluoromethyl) cyclopropyl) -3- (1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine (20 mg, 0.044 mmol, 74.71%) as a yellow solid. LC/MS (ESI) : m/z 458 [M+H] ⁺.

Step 4. (R) -4- (7- (1- (fluoromethyl) cyclopropyl) -3- (1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine

To a solution of (3R) -4- (7- (1- (fluoromethyl) cyclopropyl) -3- (1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine (50 mg, 0.109 mmol) in MeOH (5 mL) was added TsOH (19 mg, 0.109 mmol) . The mixture was stirred at room temperature for 1 h. LC-MS showed the reaction was completed. The mixture was poured into saturated NaHCO₃ (20 mL) and extracted with EA (20 mL *3) . The combined organic phase was washed with brine (20 mL) , dried over anhydrous Na₂SO₄, filtered and concentrated in vacuo. The residue was purified by Prep-HPLC (C₁₈, 10-95%, MeOH in H₂O with 0.1%HCOOH) to afford (R) -4- (7- (1- (fluoromethyl) cyclopropyl) -3- (1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine (10 mg, 0.027 mmol, 24.50%) . LC/MS (ESI) : m/z 374 [M+H] ⁺. ¹H NMR (400 MHz, DMSO) δ 13.46 (d, J = 169.5 Hz, 1H) , 7.78 (d, J = 88.4 Hz, 1H) , 7.39 (d, J = 1.9 Hz, 1H) , 7.24 (s, 1H) , 4.63 (s, 1H) , 4.18 (d, J = 13.5 Hz, 1H) , 4.04 (d, J = 11.0 Hz, 1H) , 3.82 (d, J = 11.3 Hz, 1H) , 3.72 (dd, J = 11.4, 2.8 Hz, 1H) , 3.57 (td, J = 11.7, 2.8 Hz, 1H) , 3.26 (d, J = 12.1 Hz, 1H) , 2.85 –2.58 (m, 4H) , 2.12 (dd, J = 8.5, 2.9 Hz, 1H) , 1.88 (d, J = 9.8 Hz, 1H) , 1.24 (d, J = 6.6 Hz, 3H) .

Example 32

Synthesis of (3R) -4- [7- (1-ethyl-1H-1, 2, 3-triazol-5-yl) -3- (1H-pyrazol-5-yl) - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] -3-methylmorpholine

Step 1. (3R) -4- [3-chloro-7- (1-ethyl-1H-1, 2, 3-triazol-5-yl) - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] -3-methylmorpholine

A mixture of (3R) -4- {3, 7-dichloro- [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine (200 mg, 0.657 mmol) , 1-ethyl-1H-1, 2, 3-triazole (383.12 mg, 3.945 mmol) , Tetramethylammonium acetate (262.70 mg, 1.972 mmol) and Pd (PPh₃) ₂Cl₂ (92.29 mg, 0.131 mmol) in DMA (3 mL) was stirred at 140℃ for 8 hrs under nitrogen atmosphere. The reaction mixture was diluted with water and extracted with ethyl acetate. The organic layer was washed with brine, dried over Na₂SO₄, filtered and concentrated in vacuo. The residue was purified on flash column chromatography (Silica, 0～100%ethyl acetate in petroleum ether) to give the title product (3R) -4- [3-chloro-7- (1-ethyl-1H-1, 2, 3-triazol-5-yl) - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] -3-methylmorpholine (118 mg, 0.323 mmol, 49.19%) as a yellow semi-solid. LC-MS (ESI+) : m/z (M+H) = 364.8, 366.

Step 2. (3R) -4- [7- (1-ethyl-1H-1, 2, 3-triazol-5-yl) -3- [1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] -3-methylmorpholine

A mixture of (3R) -4- [3-chloro-7- (1-ethyl-1H-1, 2, 3-triazol-5-yl) - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] -3-methylmorpholine (118 mg, 0.323 mmol) , 1- (oxan-2-yl) -5- (tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole (179.92 mg, 0.647 mmol) , Pd (PPh₃) ₄ (74.74 mg, 0.065 mmol) and K₂CO₃ (3 mL, 6.000 mmol) in dioxane (15 mL) and water (3 mL) was stirred overnight at 100℃ under nitrogen atmosphere. The reaction mixture was diluted with water and extracted with Ethyl Acetate. The combined organic layer was washed with brine, dried over Na₂SO₄, filtered and concentrated in vacuo. The residue was purified on flash column chromatography (Silica, 0～100%Ethyl Acetate in petroleum ether) to give the title product (3R) -4- [7- (1-ethyl-1H-1, 2, 3-triazol-5-yl) -3- [1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] -3-methylmorpholine (49 mg, 0.102 mmol, 31.53%) as a yellow oil. LC-MS (ESI+) : m/z (M+H) =480.

Step 3. (3R) -4- [7- (1-ethyl-1H-1, 2, 3-triazol-5-yl) -3- (1H-pyrazol-5-yl) - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] -3-methylmorpholine

To a solution of (3R) -4- [7- (1-ethyl-1H-1, 2, 3-triazol-5-yl) -3- [1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] -3-methylmorpholine (49 mg, 0.102 mmol) in DCM (2 mL) was added TFA (2 mL) and the resulting mixture was stirred for 1 h at ambient temperature. The mixture was concentrated and basified with saturated ammonium. The mixture was concentrated and the residue was purified on flash column chromatography (Silica, 0～10%MeOH in DCM) and Prep-HPLC (C18, 10-95%, acetonitrile in water with 0.1%formic acid) to give the title product (3R) -4- [7- (1-ethyl-1H-1, 2, 3-triazol-5-yl) -3- (1H-pyrazol-5-yl) - [1, 2] thiazolo [4, 5- b] pyridin-5-yl] -3-methylmorpholine (6.5 mg, 0.016 mmol, 16.08%) . LC-MS (ESI+) : m/z (M+H) = 396.8.

¹H NMR (400 MHz, DMSO) δ 13.53 (d, J = 177.6 Hz, 1H) , 8.22 (s, 1H) , 7.82 (d, J = 89.9 Hz, 1H) , 7.45 (d, J = 16.2 Hz, 2H) , 4.62 –4.47 (m, 3H) , 4.19 (d, J =12.2 Hz, 1H) , 4.05 (d, J = 10.3 Hz, 1H) , 3.83 (d, J = 11.2 Hz, 1H) , 3.73 (d, J = 11.1 Hz, 1H) , 3.59 (t, J = 10.7 Hz, 1H) , 3.30 –3.23 (m, 1H) , 1.38 (t, J = 7.2 Hz, 3H) , 1.26 (d, J = 6.4 Hz, 3H) .

Example 33

Synthesis of (3R) -3-methyl-4- [7- (4-methyl-1H-1, 2, 3-triazol-1-yl) -3- (3-methyl-1H-pyrazol-5-yl) - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] morpholine

Step 1. (3R) -4- {7-azido-3- [ (4-methoxyphenyl) methoxy] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine

To a suspension of (3R) -4- {7-chloro-3- [ (4-methoxyphenyl) methoxy] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine (300 mg, 0.739 mmol) in DMSO (5 mL) was added N₃Na (157 mg, 1.478 mmol) and Na₂CO₃ (157 mg, 1.48 mmol) After completion of the addition , the mixture was heated at 80℃ for 3 days. LC-MS showed the reaction was completed. The reaction was diluted with EA and water. The two phases were separated. The organic phase was washed with brine, dried over anhydrous Na₂SO₄, filtered and concentrated in vacuo to give crude product. The residue was purified by flash column chromatography (Silica gel, 0～100%, ethyl acetate in petroleum ether) to give (3R) -4- {7-azido-3- [ (4-methoxyphenyl) methoxy] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine (120 mg, 0.291 mmol, 39.36%) as a white solid. LC/MS (ESI) : m/z 413 (M+H) ⁺

Step 2. (3R) -4- {3- [ (4-methoxyphenyl) methoxy] -7- (4-methyl-1H-1, 2, 3-triazol-1-yl) - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine

To a suspension of (3R) -4- {7-azido-3- [ (4-methoxyphenyl) methoxy] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine (120 mg, 0.291 mmol) in MeOH (5 mL) and H₂O (1 mL) was added trimethyl (prop-1-yn-1-yl) silane (0.087 mL, 0.582 mmol) , copper (II) sulfate pentahydrate (0.064 mL, 0.582 mmol) and Sodium ascorbate (115 mg, 0.582 mmol) . After completion of the addition, the mixture was stirred at room temperature overnight. LC-MS showed the reaction was completed. The reaction was diluted with EA and water. The two phases were separated. The organic phase was washed with brine, dried over anhydrous Na₂SO₄, filtered and concentrated in vacuo to give crude product. The residue was purified by flash column chromatography (Silica gel, 0～100%ethyl acetate in petroleum ether) to give to give (3R) -4- {3- [ (4-methoxyphenyl) methoxy] -7- (4-methyl-1H-1, 2, 3-triazol-1-yl) - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine (90 mg, 0.199 mmol, 68.36%) as a yellow solid. LC/MS (ESI) : m/z 453 (M+H) ⁺.

Step 3. 7- (4-methyl-1H-1, 2, 3-triazol-1-yl) -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-3-ol

To a solution of (3R) -4- {3- [ (4-methoxyphenyl) methoxy] -7- (4-methyl-1H-1, 2, 3-triazol-1-yl) - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine (90 mg, 0.199 mmol ) in TFA (3 mL ) was stirred at room temperature for1 hour. LC-MS showed the reaction was completed. The reaction was concentrated in vacuo to give crude product, which was purified by flash column chromatography (Silica gel, 0～100%ethyl acetate in petroleum ether ) to afford the 7- (4-methyl-1H-1, 2, 3-triazol-1-yl) -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-3-ol (38 mg, 0.114 mmol, 57.48%) as a white solid. LC/MS (ESI) : m/z 333 (M+H) ⁺.

Step 4. 7- (4-methyl-1H-1, 2, 3-triazol-1-yl) -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-3-yl trifluoromethanesulfonate

To a mixture of 7- (4-methyl-1H-1, 2, 3-triazol-1-yl) -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-3-ol (30 mg, 0.090 mmol ) and DIPEA (34.93 mg, 0.271 mmol ) in THF (5 mL ) was added PhNTf₂ (64.44 mg, 0.181 mmol ) . After stirred at RT for 16hs, the mixture was concentrated to dryness. The residue was purified by flash chromatography (silica gel (12 g) , 0-100%, EA in PE ) to give 7- (4-methyl-1H-1, 2, 3-triazol-1-yl) -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-3-yl trifluoromethanesulfonate (35 mg, 0.075 mmol, 83.49%) as a yellow solid. LC/MS (ESI) m/z: 465.5 (M+1) ⁺.

Step 5. (3R) -3-methyl-4- {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -7- (4-methyl- 1H-1, 2, 3-triazol-1-yl) - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} morpholine

To a mixture of 7- (4-methyl-1H-1, 2, 3-triazol-1-yl) -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-3-yl trifluoromethanesulfonate (35 mg, 0.075 mmol) , (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) boronic acid (37.35 mg, 0.226 mmol) and K₂CO₃ (2M in H2O, 0.188 mL, 0.377 mmol) in dioxane (1.5 mL) was added Pd (dppf) Cl₂ (11.03 mg, 0.015 mmol) . The mixture was stirred at 100℃ for 16 hrs. The mixture was filtered and concentrated to dryness. The residue was purified by flash chromatography (silica gel (12 g) , 0-100%, EA in PE) to give (3R) -3-methyl-4- {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -7- (4-methyl-1H-1, 2, 3-triazol-1-yl) - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} morpholine (32 mg, 0.067 mmol, 88.36%) as a yellow oil. LC/MS (ESI) m/z: 481.6 (M+1) ⁺.

Step 6. (3R) -3-methyl-4- [7- (4-methyl-1H-1, 2, 3-triazol-1-yl) -3- (3-methyl-1H-pyrazol-5-yl) - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] morpholine

To a mixture of (3R) -3-methyl-4- {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -7- (4-methyl-1H-1, 2, 3-triazol-1-yl) - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} morpholine (32 mg, 0.067 mmol) in DCM (0.5 mL) was added HCl (4M in dioxane, 1 mL) . After stirred at rt for 1 hr, the mixture was concentrated to dryness. Then the crude product was purified by Prep-HPLC (C18, 10-95%, MeCN in H₂O with 0.1%HCOOH) to give (3R) -3-methyl-4- [7- (4-methyl-1H-1, 2, 3-triazol-1-yl) -3- (3-methyl-1H-pyrazol-5-yl) - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] morpholine (6 mg, 0.015 mmol, 22.73%) . LC/MS (ESI) m/z: 397.5 (M+1) ⁺.

¹HNMR (400 MHz, DMSO) δ 13.10 (d, J = 117.1 Hz, 1H) , 9.19 (s, 1H) , 7.70 (s, 1H) , 7.13 (s, 1H) , 4.61 (dd, J = 13.1, 6.9 Hz, 1H) , 4.27 –4.15 (m, 1H) , 4.07 (dd, J = 9.1, 3.7 Hz, 1H) , 3.86 (dd, J = 14.2, 3.0 Hz, 1H) , 3.75 (dd, J = 11.7, 1.3 Hz, 1H) , 3.60 (td, J = 11.8, 2.4 Hz, 1H) , 3.28 (s, 1H) , 2.42 (s, 3H) , 2.31 (s, 3H) , 1.28 (d, J = 6.3 Hz, 3H) .

Example 35

Synthesis of methyl ( { [3- (3-methyl-1H-pyrazol-5-yl) -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl] imino} ) phenyl-λ^6-sulfanone

Step 1. imino (methyl) phenyl-λ^6-sulfanone

To a solution of (methylsulfanyl) benzene (1 mL, 8.05 mmol) in MeOH (80 mL) were added (acetyloxy) (phenyl) -λ^3-iodanyl acetate (3.5 mL, 18.52 mmol) , diammonium carbonate (1 g, 12.08 mmol) at room temperature. The reaction was stirred at room temperature under O₂ atmosphere for 5 minutes. LC-MS showed the reaction was completed. The reaction mixture was poured into H₂O (20 mL) and extracted with EA (20 mL×3) . The combined organic phase was washed with brine (20 mL) , dried over anhydrous Na₂SO₄, filtered and concentrated to dryness. The residue was purified by flash chromatography (silica gel (10g) , 0-100%, EA in PE) to afford the desired product as a light yellow solid imino (methyl) phenyl-λ^6-sulfanone (1.1 g, yield: 88%) . LC/MS (ESI) : m/z 156.1 [M+H] ⁺.

Step 2. (3S) -1- [3- (3-methyl-1H-pyrazol-5-yl) -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl] pyrrolidin-3-ol

To a solution of imino (methyl) phenyl-λ^6-sulfanone (150 mg, 0.97 mmol) in dioxane (10 mL) were added (3R) -4- {7-chloro-3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine (419 mg, 0.97 mmol) , Xantphos (112 mg, 0.19 mmol) , Pd₂ (dba) ₃ (89 mg, 0.10 mmol) , CS₂CO₃ (945 mg, 2.90 mmol) . The reaction was stirred at 100℃ overnight under N₂. The mixture was added H₂O (10 mL) and extracted with EtOAc (6 mL x 3) . The combined organic phase was washed with brine (20 mL) , dried over anhydrous Na₂SO₄, filtered and concentrated to dryness. The residue was purified by flash chromatography to give methyl ( {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} imino) phenyl-λ^6-sulfanone (120 mg, yield: 22%) . LC-MS (ESI) : m/z 534.6 [M+H] ⁺.

Step 3. methyl ( { [3- (3-methyl-1H-pyrazol-5-yl) -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl] imino} ) phenyl-λ^6-sulfanone

To a solution of methyl ( {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} imino) phenyl-λ^6-sulfanone (60 mg, 0.109 mmol) in DCM (1 mL) was added HCl/dioxane (2 mL) . The mixture was stirred at room temperature for 1 h. LC-MS showed the reaction was completed. The reaction mixture was concentrated under vacuo. The residue was purified by SFC to afford 35a (14.4 mg, 0.031 mmol, 28.31%) and 35b (4 mg, yield: 7%) . LC/MS (ESI) m/z: 468.8 [M+H] ⁺.

35a:

¹H NMR (400 MHz, DMSO-d6) δ 8.08 (t, J = 28.4 Hz, 2H) , 7.71 (dt, J = 15.0, 7.3 Hz, 3H) , 7.03 (s, 1H) , 6.26 (s, 1H) , 4.13 –3.86 (m, 2H) , 3.84 –3.61 (m, 5H) , 3.55 (dd, J = 13.1, 10.8 Hz, 1H) , 3.51 –3.41 (m, 1H) , 2.99 (td, J = 12.6, 3.5 Hz, 1H) , 2.27 (s, 3H) , 1.11 (d, J = 6.6 Hz, 3H) .

35b:

¹H NMR (400 MHz, DMSO-d6) δ 8.06 –7.97 (m, 2H) , 7.76 –7.60 (m, 3H) , 7.02 (s, 1H) , 6.16 (s, 1H) , 3.97 –3.82 (m, 3H) , 3.69 (s, 3H) , 3.67 –3.56 (m, 2H) , 3.45 –3.42 (m, 1H) , 2.99 (td, J = 12.8, 3.7 Hz, 1H) , 2.27 (s, 3H) , 0.73 (d, J = 6.6 Hz, 3H) .

Example 36

Synthesis of (R) -3-methyl-4- (3- (3-methyl-1H-pyrazol-5-yl) -7-morpholinoisothiazolo [4, 5-b] pyridin-5-yl) morpholine

Step 1. (3R) -3-methyl-4- (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -7-morpholinoisothiazolo [4, 5-b] pyridin-5-yl) morpholine

To a solution of (3R) -4- (7-chloro-3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine (60 mg, 0.14 mmol) in Sulfolane (2 mL) were added morpholine (0.02 mL, 0.28 mmol) , DIPEA (0.12 mL, 0.69 mmol) and KF (8 mg, 0.14 mmol) . The reaction was stirred at 120℃ overnight under nitrogen atmosphere. LC-MS showed the reaction was completed. The mixture was poured into water and extracted with EA (20 mL x 3) . The combined organic phase was washed with brine (20 mL) , dried over anhydrous Na₂SO₄, filtered and concentrated in vacuo. The residue was purified by Prep-TLC (PE: EA = 1: 1, V/V) to afford (3R) -3-methyl-4- (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -7-morpholinoisothiazolo [4, 5-b] pyridin-5-yl) morpholine (50 mg, yield: 75%) as a green solid. LC/MS (ESI) : m/z 485.6 (M+H) ⁺.

Step 2. (R) -3-methyl-4- (3- (3-methyl-1H-pyrazol-5-yl) -7-morpholinoisothiazolo [4, 5-b] pyridin-5-yl) morpholine

To a solution of (3R) -3-methyl-4- (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -7-morpholinoisothiazolo [4, 5-b] pyridin-5-yl) morpholine (35 mg, 0.07 mmol) in DCM (1 mL) was added HCl (4 M in dioxane, 2 mL) . The mixture was stirred at room temperature for 0.5 h. LC-MS showed the reaction was completed. The reaction mixture was concentrated in vacuo. The residue was purified by Prep-HPLC (C₁₈, 10-95%, MeOH in H₂O with 0.1%HCOOH) to afford (R) -3-methyl-4- (3-(3-methyl-1H-pyrazol-5-yl) -7-morpholinoisothiazolo [4, 5-b] pyridin-5-yl) morpholine (5 mg, yield: 18%) . LC/MS (ESI) : m/z 401 [M+H] ⁺.

¹H NMR (400 MHz, DMSO-d6) δ 13.01 (s, 1H) , 7.11 (s, 1H) , 6.36 (s, 1H) , 4.49 (d, J = 6.3 Hz, 1H) , 4.01 (dd, J = 16.5, 8.1 Hz, 2H) , 3.80 (dd, J = 10.7, 6.3 Hz, 5H) , 3.69 (dd, J = 11.3, 2.7 Hz, 1H) , 3.57 –3.51 (m, 1H) , 3.41 (d, J = 2.4 Hz, 4H) , 3.20 (td, J = 12.8, 3.7 Hz, 1H) , 2.29 (s, 3H) , 1.20 (d, J = 6.6 Hz, 3H) .

Example 37

Synthesis of (3R, 3'R) -4, 4'- (3- (3-methyl-1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridine-5, 7-diyl) bis (3-methylmorpholine)

Step 1. (3R, 3'R) -4, 4'- (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5- yl) isothiazolo [4, 5-b] pyridine-5, 7-diyl) bis (3-methylmorpholine)

To a mixture of (3R) -4- {7-chloro-3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine (80 mg, 0.18 mmol) , (3R) -3-methylmorpholine (37 mg, 0.37 mmol) and CS₂CO₃ (120 mg, 0.37 mmol) in N, N-dimethylformamide (2 mL) were added RuPhos Pd G3 (15 mg, 0.02 mmol) , RuPhos (17 mg, 0.04 mmol) . The mixture was stirred at 80℃ for 16 hrs under N₂. LC-MS showed the reaction was completed. The mixture was poured into H₂O (50 mL) and extracted with EA (50 mL x 3) . The combined organic phase was washed brine (50 mL) , dried over anhydrous Na₂SO₄, filtered and concentrated. The residue was purified by flash chromatography (silica gel (12 g) , 0-100%, EA in PE) to give (3R) -3-methyl-4- {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -7- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} morpholine (70 mg, yield: 76%) as a yellow oil. LC/MS (ESI) : m/z 498.65 [M+H] ⁺.

Step 2. (3R, 3'R) -4, 4'- (3- (3-methyl-1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridine-5, 7-diyl) bis (3-methylmorpholine)

To a mixture of (3R) -3-methyl-4- {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -7- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} morpholine (70 mg, 0.14 mmol) in DCM (1 mL) was added HCl/dioxane (1 mL) . The mixture was stirred at rt for 1 h. LC-MS showed the reaction was completed. The mixture was concentrated to dryness. Then the crude product was purified by prep-HPLC (C₁₈, 10-95%, MeCN in H₂O with 0.1%HCOOH) to give (3R) -3-methyl-4- [3- (3-methyl-1H-pyrazol-5-yl) -7- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] morpholine (23 mg, yield: 39%) . LC/MS (ESI) : m/z 414.5 [M+H] ⁺.

¹H NMR (400 MHz, DMSO-d6) δ 13.00 (s, 1H) , 7.10 (s, 1H) , 6.29 (s, 1H) , 4.48 (d, J = 5.4 Hz, 1H) , 4.11 –3.94 (m, 4H) , 3.78 (d, J = 10.0 Hz, 3H) , 3.72 –3.50 (m, 3H) , 3.42 –3.38 (m, 2H) , 3.19 (td, J = 12.8, 3.7 Hz, 1H) , 2.29 (s, 3H) , 1.21 (d, J =6.6 Hz, 3H) , 1.13 (d, J = 6.6 Hz, 3H) .

Example 38

Synthesis of (S) -1- (3- (3-methyl-1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) pyrrolidin-3-ol

Step 1. (3S) -1- (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) pyrrolidin-3-ol

To a mixture of (3R) -4- {7-chloro-3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine (80 mg, 0.18 mmol) in Sulfolane (2 mL) were added (3S) -pyrrolidin-3-ol (19 mg, 0.22 mmol) and DIEA (0.2 mL, 0.92 mmol) and KF (11 mg, 0.18 mmol) . The mixture was stirred at 120℃ for 15 h. LC-MS showed the reaction was completed. The mixture was poured into water (30 mL) and extracted with EA (30 mL x 3) . The combined organic phase was washed with brine (30 mL) , dried over anhydrous Na₂SO₄, filtered and concentrated in vacuo. The residue was purified by Prep-TLC (PE: EA = 1: 1, V/V) to give (3S) -1- (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) pyrrolidin-3-ol (30 mg, yield: 34%) as a yellow solid. LC/MS (ESI) : m/z 485.8 [M+H] ⁺.

Step 2. (S) -1- (3- (3-methyl-1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) pyrrolidin-3-ol

To a solution of (3S) -1- (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) pyrrolidin-3-ol (30 mg, 0.06 mmol) in DCM (1 mL) was added HCl (4 M in dioxane, 1 mL) . The mixture was stirred at rt for 1 h. LC-MS showed the reaction was completed. The reaction mixture was concentrated under reduced pressure. The residue was purified by Prep-HPLC (C₁₈, 10-95%, MeOH in H₂O with 0.1%HCOOH) to give (S) -1- (3- (3-methyl-1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) pyrrolidin-3-ol (10 mg, yield: 20%) . LC/MS (ESI) : m/z 401.3 [M+H] ⁺.

¹H NMR (400 MHz, DMSO-d6) δ 7.08 (s, 1H) , 5.80 (s, 1H) , 5.13 (d, J = 2.6 Hz, 1H) , 4.45 (s, 2H) , 4.05 –3.86 (m, 2H) , 3.82 –3.65 (m, 5H) , 3.57 –3.42 (m, 2H) , 3.15 (td, J = 12.6, 3.8 Hz, 1H) , 2.30 (d, J = 19.1 Hz, 3H) , 2.09 (ddd, J = 12.9, 8.6, 4.5 Hz, 1H) , 1.98 (d, J = 3.0 Hz, 1H) , 1.17 (d, J = 6.6 Hz, 3H) .

Example 39

Synthesis of (R) -1- (3- (3-methyl-1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) pyrrolidin-3-ol

Step 1. (3R) -1- (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) pyrrolidin-3-ol (ZYM-N191165-196)

To a mixture of (3R) -4- {7-chloro-3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine (80 mg, 0.18 mmol) in Sulfolane (2 mL) were added (R) -pyrrolidin-3-ol (19 mg, 0.22 mmol) and DIEA (0.2 mL, 0.92 mmol) and KF (11 mg, 0.18 mmol) . The mixture was stirred at 120℃ for 15 h. LC-MS showed the reaction was completed. The mixture was poured into water (50 mL) and extracted with EA (50 mL x 3) . The combined organic phase was washed with brine (50 mL) , dried over anhydrous Na₂SO₄, filtered and concentrated in vacuo. The residue was purified by Prep-TLC (PE: EA = 1: 1, V/V) to give (3R) -1- (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3- methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) pyrrolidin-3-ol (30 mg, yield: 34%) as a yellow solid. LC/MS (ESI) : m/z 485.8 [M+H] ⁺.

Step 2. (R) -1- (3- (3-methyl-1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) pyrrolidin-3-ol (KYH-N211770-025-1)

To a solution of (3R) -1- (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) pyrrolidin-3-ol (30 mg, 0.06 mmol) in DCM (1 mL) was added HCl (4 M in dioxane, 1 mL) . The mixture was stirred at rt for 1 h. LC-MS showed the reaction was completed. The reaction mixture was concentrated under reduced pressure. The residue was purified by Prep-HPLC (C₁₈, 10-95%, MeOH in H₂O with 0.1%HCOOH) to give (R) -1- (3- (3-methyl-1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) pyrrolidin-3-ol (10 mg, yield: 20%) . LC/MS (ESI) : m/z 401.3 [M+H] ⁺.

¹H NMR (400 MHz, DMSO-d6) δ 12.89 (s, 1H) , 7.08 (s, 1H) , 5.80 (s, 1H) , 5.14 (d, J = 3.3 Hz, 1H) , 4.45 (s, 2H) , 3.97 (t, J = 10.9 Hz, 2H) , 3.80 –3.75 (m, 2H) , 3.70 (dd, J = 11.4, 8.8 Hz, 3H) , 3.57 –3.50 (m, 2H) , 3.20 –3.12 (m, 1H) , 2.28 (s, 3H) , 2.09 (td, J = 8.6, 4.3 Hz, 1H) , 1.99 (s, 1H) , 1.18 (d, J = 6.6 Hz, 3H) .

Example 40

Synthesis of (R) -4- (7- (3, 3-difluoroazetidin-1-yl) -3- (3-methyl-1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine

Step 1. (3R) -4- (7- (3, 3-difluoroazetidin-1-yl) -3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine

To a solution of 3, 3-difluoroazetidine (21 mg, 0.23 mmol) , (3R) -4- {7-chloro-3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine (100 mg, 0.23 mmol) and Ruphos (12 mg, 0.05 mmol) in DMF (1.5 mL) was added RuPhos-Pd-G₃ (20 mg, 0.05 mmol) at 25℃. The reaction mixture was stirred at 80℃ for 16 h under N₂ atmosphere. LC-MS showed the reaction was completed. The reaction solution was diluted with H₂O (20 mL) , extracted with EA (20 mL×3) . The combined organic layer was washed with water and brine (20 mL) , dried over anhydrous Na₂SO₄, filtered and concentrated in vacuum. The residue was purified by flash chromatography (silica gel, 0 ～ 100 %, ethyl acetate in petroleum ether) to afford (3R) -4- [7- (3, 3-difluoroazetidin-1-yl) -3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] -3-methylmorpholine (45 mg, yield: 40%) as a colorless sticky oil. LC/MS (ESI) : m/z 491 [M+1] ⁺.

Step 2. (R) -4- (7- (3, 3-difluoroazetidin-1-yl) -3- (3-methyl-1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine

To a mixture of (R) -4- (7- (3, 3-difluoroazetidin-1-yl) -3- (3-methyl-1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine (50 mg, 0.10 mmol) in DCM (1 mL) was added HCl (4 M in dioxane, 3 mL) . The mixture was stirred at rt for 1 h. LC-MS showed the reaction was completed. The mixture was concentrated to dryness. The crude product was purified by Prep-HPLC (C₁₈, 10-95%, MeCN in H₂O with 0.1%HCOOH) to give (R) -4- (7- (3, 3-difluoroazetidin-1-yl) -3- (3-methyl-1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine (9 mg, yield: 22%) as a yellow solid. LC/MS (ESI) m/z: 407 [M+1] ⁺.

¹H NMR (400 MHz, DMSO-d6) δ 7.11 (s, 1H) , 6.04 (s, 1H) , 4.66 (t, J = 12.2 Hz, 4H) , 4.47 (d, J = 4.6 Hz, 1H) , 4.01 (d, J = 11.0 Hz, 2H) , 3.79 (d, J = 11.2 Hz, 1H) , 3.68 (dd, J = 11.3, 2.8 Hz, 1H) , 3.52 (dt, J = 11.6, 6.0 Hz, 1H) , 3.18 (dd, J = 12.5, 9.0 Hz, 1H) , 2.29 (s, 3H) , 1.20 (d, J = 6.6 Hz, 3H) .

Example 41

Synthesis of ethyl ( (3- (3-methyl-1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) (phenyl) -λ^6-sulfanone

Step 1. ethyl ( (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) - 3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) (phenyl) -λ^6-sulfanone

To a solution of (3R) -4- {7-chloro-3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine (150 mg, 0.35 mmol) in dioxane (3 mL) were added ethyl (imino) phenyl-λ^6-sulfanone (70 mg, 0.42 mmol) , Cs₂CO₃ (338 mg, 1.0 mmol) , Pd₂ (dba) ₃ (32 mg, 0.035 mmol) and Xantphos (40 mg, 0.07 mmol) . The reaction was stirred at 100℃ overnight under nitrogen atmosphere. LC-MS showed the reaction was completed. The reaction was filtered and concentrated in vacuo. The residue was purified by flash column chromatography (Silica gel, 0～100%ethyl acetate in petroleum ether) to give ethyl ( (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) (phenyl) -λ^6-sulfanone (190 mg, yield: 97%) as a yellow solid. LC/MS (ESI) : m/z 567.7 (M+H) ⁺.

Step 2. ethyl ( (3- (3-methyl-1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) (phenyl) -λ^6-sulfanone

To a solution of ethyl ( (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) (phenyl) -λ^6-sulfanone (190 mg, 0.34 mmol) in DCM (4 mL) was added HCl (4 M in dioxane, 4 mL) . The mixture was stirred at room temperature for 1 h. LC-MS showed the reaction was completed. The reaction mixture was concentrated in vacuo. The residue was purified by SFC to afford 41a (22 mg, yield: 13%) and 41b (15 mg, yield: 9%) . LC/MS (ESI) : m/z 483.6 (M+H) ⁺.

41a:

¹H NMR (400 MHz, DMSO-d6) δ 8.00 –7.87 (m, 2H) , 7.75 (t, J = 7.4 Hz, 1H) , 7.67 (t, J = 7.5 Hz, 2H) , 7.02 (s, 1H) , 6.16 (s, 1H) , 3.94 (d, J = 7.8 Hz, 1H) , 3.85 (d, J = 12.0 Hz, 2H) , 3.81 –3.73 (m, 2H) , 3.63 (dt, J = 11.4, 7.1 Hz, 2H) , 3.44 (dd, J =11.8, 9.0 Hz, 1H) , 3.07 –2.89 (m, 1H) , 2.27 (s, 3H) , 1.25 (t, J = 7.3 Hz, 3H) , 0.71 (d, J = 6.6 Hz, 3H) .

41b:

¹H NMR (400 MHz, DMSO-d6) δ 12.98 (s, 1H) , 8.04 –7.90 (m, 2H) , 7.75 (d, J = 7.3 Hz, 1H) , 7.68 (t, J = 7.6 Hz, 2H) , 7.04 (s, 1H) , 6.27 (s, 1H) , 4.02 (d, J = 5.2 Hz, 1H) , 3.93 (dd, J = 11.3, 3.3 Hz, 1H) , 3.81 –3.74 (m, 3H) , 3.71 (d, J = 11.2 Hz, 1H) , 3.56 (dd, J = 11.4, 2.8 Hz, 1H) , 3.46 (td, J = 11.7, 2.8 Hz, 1H) , 2.98 (td, J = 12.6, 3.7 Hz, 1H) , 2.27 (s, 3H) , 1.23 (t, J = 7.3 Hz, 3H) , 1.10 (d, J = 6.6 Hz, 3H) .

Example 42

Synthesis of methyl ( (3- (3-methyl-1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) (pyridin-2-yl) -λ^6-sulfanone

Step 1. imino (methyl) (pyridin-2-yl) -λ^6-sulfanone

To a suspension of 2- (methylsulfanyl) pyridine (1 g, 8.0 mmol) in MeOH (10 mL) was added (Diacetoxyiodo ) benzene (6 g, 18.40 mmol) and Ammonium carbonate (1 g, 12.00 mmol) . The mixture was stirred at room temperature for 10 minutes under air. LC-MS showed the reaction was completed. The reaction was poured into water and extracted with EA (50 mL x 3) . The combined organic phase was washed with brine (50 mL) , dried over anhydrous Na₂SO₄, filtered and concentrated in vacuo. The residue was purified by flash column chromatography (Silica gel, 0～100%, ethyl acetate in petroleum ether) to give imino (methyl) (pyridin-2-yl) -λ^6-sulfanone (520 mg, yield: 42%) as a yellow solid. LC/MS (ESI) : m/z 157.2 (M+H) ⁺.

Step 2. methyl ( (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) (pyridin-2-yl) -λ^6-sulfanone

To a solution of (3R) -4- {7-chloro-3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine (200 mg, 0.46 mmol) in dioxane (5 mL) were added imino (methyl) (pyridin-2-yl) -λ^6-sulfanone (86 mg, 0.55 mmol) , Cs₂CO₃ (450 mg, 1.4 mmol) , Pd₂ (dba) ₃ (42 mg, 0.05mmol) and Xantphos (53 mg, 0.09 mmol) . The reaction was stirred at 100℃ overnight under nitrogen atmosphere. LC-MS showed the reaction was completed. The reaction was cooled to rt, filtered and concentrated in vacuo. The residue was purified by flash column chromatography (Silica gel, 0～100%, ethyl acetate in petroleum ether) to give methyl ( (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) (pyridin-2-yl) -λ^6-sulfanone (200 mg, yield: 78%) as a yellow solid. LC/MS (ESI) : m/z 554.2 (M+H) ⁺.

Step 3. methyl ( (3- (3-methyl-1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) (pyridin-2-yl) -λ^6-sulfanone

To a solution of methyl ( (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) (pyridin-2-yl) -λ^6-sulfanone (200 mg, 0.36 mmol) in DCM (3 mL) was added HCl/dioxane (3 mL) . The mixture was stirred at room temperature for 1 h. LC-MS showed the reaction was completed. The reaction mixture was concentrated in vacuo. The residue was purified by Prep-HPLC (C₁₈, 10-95%, MeOH in H₂O with 0.1%HCOOH) to give crude product. The crude product was purified by SFC to afford 42a (18 mg, yield: 11%) and 42b (26 mg, yield: 16%) as a white solid. LC/MS (ESI) : m/z 470.3 (M+H) ⁺.

42a:

¹H NMR (400 MHz, DMSO-d6) δ 12.95 (d, J = 101.2 Hz, 1H) , 8.80 (d, J =4.1 Hz, 1H) , 8.30 (t, J = 26.0 Hz, 1H) , 8.20 (t, J = 7.7 Hz, 1H) , 7.76 (dd, J = 7.4, 4.8 Hz, 1H) , 7.03 (s, 1H) , 6.33 (s, 1H) , 4.14 (s, 1H) , 3.96 (d, J = 9.0 Hz, 1H) , 3.82 (d, J =11.9 Hz, 1H) , 3.74 (d, J = 11.5 Hz, 1H) , 3.71 (s, 3H) , 3.60 (d, J = 10.8 Hz, 1H) , 3.49 (t, J = 10.6 Hz, 1H) , 3.02 (t, J = 11.1 Hz, 1H) , 2.26 (s, 3H) , 1.12 (d, J = 6.6 Hz, 3H) .

42b:

¹H NMR (400 MHz, DMSO-d6) δ 12.94 (d, J = 100.0 Hz, 1H) , 8.78 (d, J =3.8 Hz, 1H) , 8.40 (d, J = 7.9 Hz, 1H) , 8.21 (td, J = 7.8, 1.7 Hz, 1H) , 7.94 –7.58 (m, 1H) , 7.01 (s, 1H) , 6.26 (s, 1H) , 4.07 –3.92 (m, 2H) , 3.88 (d, J = 12.5 Hz, 1H) , 3.78 –3.61 (m, 5H) , 3.47 (td, J = 11.6, 2.6 Hz, 1H) , 3.10 –2.92 (m, 1H) , 2.26 (s, 3H) , 0.80 (dd, J = 19.5, 6.9 Hz, 3H) .

Example 43

Synthesis of (S) -3-methyl-4- (3- (3-methyl-1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) morpholine

Step 1. (3S) -3-methyl-4- (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) morpholine

To a solution of (3R) -4- {7-chloro-3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine (100 mg, 0.23 mmol) in DMF (5 mL) were added (3S) -3-methylmorpholine (47 mg, 0.46 mmol) , Cs₂CO₃ (150 mg, 0.46 mmol) , RuPhos (22 mg, 0.05 mmol) and RuPhos-Pd-G₃ (19 mg, 0.02 mmol) . The reaction was stirred at 80℃ overnight under nitrogen atmosphere. LC-MS showed the reaction was completed. The mixture was poured into water (30 mL) and extracted with EA (30 mL x 3) . The combined organic phase was washed with brine (30 mL) , dried over anhydrous Na₂SO₄, filtered and concentrated in vacuo. The residue was purified by flash column chromatography (Silica gel, 0～100%, ethyl acetate in petroleum ether ) to give (3S) -3-methyl-4- (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) morpholine (70 mg, yield: 61%) as a yellow solid. LC/MS (ESI) : m/z 499.7 (M+H) ⁺.

Step 2. (S) -3-methyl-4- (3- (3-methyl-1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) morpholine

To a solution of (3S) -3-methyl-4- (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) morpholine (80 mg, 0.16 mmol) in DCM (2 mL) was added HCl (4 M in dioxane, 2 mL) . The mixture was stirred at room temperature for 1 h. LC-MS showed the reaction was completed. The reaction was concentrated in vacuo. The residue was purified by Prep-HPLC (C₁₈, 10-95%, MeCN in H₂O with 0.1%CHOOH) to afford (S) -3-methyl-4- (3- (3-methyl-1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) morpholine (22 mg, yield: 33%) . LC/MS (ESI) : m/z 415.6 (M+H) ⁺.

¹H NMR (400 MHz, DMSO-d6) δ 13.02 (s, 1H) , 7.10 (s, 1H) , 6.30 (s, 1H) , 4.47 (d, J = 5.4 Hz, 1H) , 4.01 (dd, J = 13.0, 5.6 Hz, 4H) , 3.79 (d, J = 10.7 Hz, 3H) , 3.70 (dd, J = 11.3, 2.6 Hz, 1H) , 3.63 (dt, J = 11.4, 7.8 Hz, 1H) , 3.59 –3.49 (m, 1H) , 3.40 (d, J = 5.5 Hz, 2H) , 3.19 (td, J = 12.7, 3.6 Hz, 1H) , 2.29 (s, 3H) , 1.15 (dd, J = 24.0, 6.6 Hz, 6H) .

Example 44

Synthesis of (R) -diethyl ( (3- (3-methyl-1H-pyrazol-5-yl) -5- (3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) -l6-sulfanone

Step 1. diethyl (imino) -l6-sulfanone

To a mixture of (ethylsulfanyl) ethane (1 g, 11.09 mmol) and Ammonium carbamate (1.7 g, 12.81 mmol) in MeOH (10 mL) was added PhI (OAc) ₂ (8.9 g, 27.63 mmol) . The mixture was stirred at 25℃ for 3 hrs. LC-MS showed the reaction was completed. The mixture was filtered and concentrated. Then the residue was purified by flash chromatography (silica gel (12 g) , 0-50%, EA in PE) to give diethyl (imino) -λ^6-sulfanone (512 mg, yield: 38%) as a colorless liquid. LC/MS (ESI) : m/z 121.2 [M+1] ⁺.

Step 2. diethyl ( (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) -l6-sulfanone

To a mixture of diethyl (imino) -λ^6-sulfanone (27 mg, 0.223 mmol) , (3R) -4- {7-chloro-3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine (80 mg, 0.18 mmol) , CS₂CO₃ (180 mg, 0.55 mmol) and XantPhos (21 mg, 0.04 mmol) in dioxane (1 mL) was added Pd₂ (dba) ₃ (17 mg, 0.02 mmol) . The mixture was stirred at 80℃ for 16 hrs under N₂. LC-MS showed the reaction was complete. The mixture was poured into H₂O (50 mL) and extracted with EA (50 mL x 3) . The combined organic phase was washed brine (50 mL) , dried over anhydrous Na₂SO₄, filtered and concentrated to dryness. The residue was purified by flash chromatography (silica gel (4 g) , 0-10%, MeOH in DCM) to give diethyl ( {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} imino) -λ^6-sulfanone (30 mg, yield: 26%) as a yellow solid. LC/MS (ESI) : m/z 518.7 [M+H] ⁺.

Step 3. (R) -diethyl ( (3- (3-methyl-1H-pyrazol-5-yl) -5- (3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) -l6-sulfanone

To a mixture of diethyl ( {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} imino) -λ^6-sulfanone (50 mg, 0.10 mmol) in DCM (1 mL) was added HCl/dioxane (1 mL) . The mixture was stirred at rt for 1h. LC-MS showed the reaction was complete. The mixture was concentrated to dryness. Then the crude product was purified by prep-HPLC (C₁₈, 10-95%, MeCN in H₂O with 0.1%HCOOH) to give diethyl ( { [3- (3-methyl-1H-pyrazol-5-yl) -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl] imino} ) -λ^6-sulfanone (27.9 mg, 0.06 mmol, 67%) . LC/MS (ESI) : m/z 434.59 [M+H] ⁺.

¹H NMR (400 MHz, DMSO-d6) δ 7.08 (s, 1H) , 6.58 (s, 1H) , 4.32 (d, J = 5.6 Hz, 1H) , 3.97 (dd, J = 27.2, 10.1 Hz, 2H) , 3.79 (d, J = 11.3 Hz, 1H) , 3.70 (dd, J = 11.3, 2.6 Hz, 1H) , 3.55 (q, J = 7.4 Hz, 5H) , 3.22 –3.11 (m, 1H) , 2.29 (s, 3H) , 1.31 (t, J = 7.3 Hz, 6H) , 1.19 (d, J = 6.6 Hz, 3H) .

Example 45

Synthesis of methyl [methyl ( {1- [3- (3-methyl-1H-pyrazol-5-yl) -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl] cyclopropyl} ) oxo-λ^6-sulfanylidene] amine

Step 1. methyl [methyl (1- {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} cyclopropyl) oxo-λ^6-sulfanylidene] amine

To a suspension of imino (methyl) (1- {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} cyclopropyl) -λ^6-sulfanone (30 mg, 0.06 mmol) in THF (3 mL) were added CH₃I (0.007 mL, 0.12 mmol) and t-BuOK (26 mg, 0.23 mmol) . After completion of the addition, the mixture was stirred at room temperature for 3 hrs. LC-MS showed the reaction was completed. The reaction was diluted with EA (50 mL) and water (50 mL) . The organic phase was washed with brine (50 mL) , dried over anhydrous Na₂SO₄, filtered and concentrated in vacuo to give crude product. The residue was purified by flash column chromatography (Silica gel, 0～100%ethyl acetate in petroleum ether) to give methyl [methyl (1- {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} cyclopropyl) oxo-λ^6-sulfanylidene] amine (30 mg, yield: 97%) as a yellow solid. LC/MS (ESI) : m/z 531 (M+H) ⁺.

Step 2. methyl [methyl ( {1- [3- (3-methyl-1H-pyrazol-5-yl) -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl] cyclopropyl} ) oxo-λ^6-sulfanylidene] amine

To a solution of methyl [methyl (1- {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7- yl} cyclopropyl) oxo-λ^6-sulfanylidene] amine (30 mg, 0.06 mmol) in DCM (2 mL) was added HCl/dioxane (2 mL) . The mixture was stirred at room temperature for 1 hour. LC-MS showed the reaction was completed. The reaction was concentrated in vacuo to give crude product, which was purified by prep-HPLC (C18, 10-95%, MeCN in H₂O with 0.1%CHOOH) to afford methyl [methyl ( {1- [3- (3-methyl-1H-pyrazol-5-yl) -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl] cyclopropyl} ) oxo-λ^6-sulfanylidene] amine (6 mg, yield: 23%) . LC/MS (ESI) : m/z 447(M+H) ⁺.

¹H NMR (400 MHz, DMSO-d6) δ 12.98 (s, 1H) , 7.43 (d, J = 5.0 Hz, 1H) , 7.09 (s, 1H) , 4.50 (s, 1H) , 4.13 (d, J = 12.6 Hz, 1H) , 4.03 (d, J = 10.9 Hz, 1H) , 3.82 (d, J = 11.4 Hz, 1H) , 3.71 (d, J = 9.2 Hz, 1H) , 3.57 (t, J = 10.4 Hz, 1H) , 3.26 –3.16 (m, 1H) , 2.96 (d, J = 9.5 Hz, 3H) , 2.69 (d, J = 6.1 Hz, 3H) , 2.30 (s, 3H) , 1.76 (d, J =10.2 Hz, 1H) , 1.72 –1.59 (m, 1H) , 1.45 (q, J = 9.7 Hz, 2H) , 1.30 –1.13 (m, 3H) .

Example 46

Synthesis of (R) -1- (3- (3-methyl-1H-pyrazol-5-yl) -5- (3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) azetidin-3-ol

Step 1. 1- (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) azetidin-3-ol

To a mixture of azetidin-3-ol (45 mg, 0.616 mmol) , (3R) -4- {7-chloro-3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine (90 mg, 0.21 mmol) and CS₂CO₃ (315 mg, 0.97 mmol) in DMF (1 mL) was added RuPhos (18 mg, 0.04 mmol) and RuPhos-Pd-G3 (18 mg, 0.02 mmol) . The mixture was stirred at 80℃ for 16 hrs under N₂. LC-MS showed the reaction was completed. The mixture was poured into H₂O (30 mL) and extracted with EA (50 mL x 3) . The combined organic phase was washed brine (30 mL) , dried over anhydrous Na₂SO₄, filtered and concentrated. The residue was purified by flash chromatography (silica gel (12 g) , 0-100%, EA in PE) to give 1- {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} azetidin-3-ol (51 mg, yield: 18%) as a yellow liquid. LC/MS (ESI) : m/z 471.2 [M+H] ⁺.

Step 2. (R) -1- (3- (3-methyl-1H-pyrazol-5-yl) -5- (3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) azetidin-3-ol

To a mixture of 1- {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} azetidin-3-ol (51 mg, 0.11 mmol) in DCM (1 mL) was added HCl/dioxane (1 mL) . The mixture was stirred at r.t for 1 h. LC-MS showed the reaction was completed. The mixture was concentrated to dryness. Then the crude product was purified by prep-HPLC (C18, 10-95%, MeCN in H₂O with 0.1%HCOOH) to give 1- [3- (3-methyl-1H-pyrazol-5-yl) -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl] azetidin-3-ol (9.3 mg, yield: 22%) . LC/MS (ESI) : m/z 387.4 [M+H] ⁺.

¹H NMR (400 MHz, DMSO-d6) δ 7.05 (s, 1H) , 5.80 (s, 1H) , 4.67 (s, 1H) , 4.42 (t, J = 7.3 Hz, 3H) , 4.04 –3.91 (m, 4H) , 3.78 (d, J = 11.3 Hz, 1H) , 3.69 (dd, J =11.4, 2.7 Hz, 1H) , 3.58 –3.45 (m, 2H) , 2.29 (s, 3H) , 1.19 (d, J = 6.6 Hz, 3H) .

Example 47

Synthesis of methyl (1-methyl-1H-pyrazol-4-yl) ( (3- (3-methyl-1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) -l6-sulfanone

Step 1. 1-methyl-4- (methylthio) -1H-pyrazole

To a mixture of 4-iodo-1-methyl-1H-pyrazole (1.5 g, 7.21 mmol) , CH₃SNa (750 mg, 7.21 mmol) and Xantphos (0.5 g, 0.86 mmol) in dioxane (15 mL) was added Pd₂ (dba) ₃ (0.4 g, 0.44 mmol) . The mixture was stirred at 80℃ for 16 hrs under N₂. LC-MS showed the reaction was completed. The mixture was poured into H₂O (50 mL) and extracted with EA (50 mL x 3) . The combined organic phase was washed brine (50 mL) , dried over anhydrous Na₂SO₄, filtered and concentrated. The residue was purified by flash chromatography (silica gel (40 g) , 0-60%, EA in PE) to give 1- methyl-4- (methylsulfanyl) -1H-pyrazole (160 mg, yield: 17%) as a yellow solid. LC/MS (ESI) : m/z 129.2 [M+H] ⁺.

Step 2. imino (methyl) (1-methyl-1H-pyrazol-4-yl) -l6-sulfanone

To a mixture of 1-methyl-4- (methylsulfanyl) -1H-pyrazole (100 mg, 0.78 mmol) and PhI (OAc) ₂ (643 mg, 0.78 mmol) in MeOH (1 mL) was added NH₂COONH₄ (155 mg, 0.78 mmol) . The mixture was stirred at 25℃ for 3hrs. LC-MS showed the reaction was completed. The mixture was poured into H₂O (50 mL) and extracted with DCM (50 mL x 3) . The mixture was filtered and concentrated to dryness to give imino (methyl) (1-methyl-1H-pyrazol-4-yl) -λ^6-sulfanone (115 mg, yield: 93%) as a white liquid. LC/MS (ESI) : m/z 160.2 [M+1] ⁺.

Step 3. methyl ( (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) (1-methyl-1H-pyrazol-4-yl) -l6-sulfanone

To a mixture of imino (methyl) (1-methyl-1H-pyrazol-4-yl) -λ^6-sulfanone (110 mg, 0.69 mmol) , (3R) -4- {7-chloro-3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine (100 mg, 0.23 mmol) , xantphos (26 mg, 0.05 mmol) and Pd₂ (dba) ₃ (21 mg, 0.02 mmol) in dioxane (1 mL) was added CS₂CO₃ (230 mg, 0.71 mmol) . The mixture was stirred at 100℃ for 16 hrs under N₂. LC-MS showed the reaction was completed. The mixture was filtered and concentrated to dryness. The residue was purified by flash chromatography (silica gel (4 g) , 0-100%, EA in PE) to give methyl ( {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} imino) (1-methyl-1H-pyrazol-4-yl) -λ^6-sulfanone (131 mg, yield: 34%) as a yellow solid. LC/MS (ESI) : m/z 556.8 [M+H] ⁺.

Step 4. methyl (1-methyl-1H-pyrazol-4-yl) ( (3- (3-methyl-1H-pyrazol-5-yl) -5- ( (R) -3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) imino) -l6-sulfanone

To a mixture of methyl ( {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} imino) (1-methyl-1H-pyrazol-4-yl) -λ^6-sulfanone (131 mg, 0.235 mmol) in DCM (2 mL) was added HCl (2 mL) . The mixture was stirred at r.t for 1 h. LC-MS showed the reaction was completed. The mixture was concentrated to dryness. Then the crude product was purified by prep-HPLC (C₁₈, 10-95%, MeCN in H₂O with 0.1%HCOOH) to give 47a (18 mg, yield: 17%) and 47b (12 mg, yield; 11%) . LC/MS (ESI) : m/z 472.8 [M+H] ⁺.

47a:

¹H NMR (400 MHz, DMSO-d6) δ 8.48 (s, 1H) , 7.96 (s, 1H) , 7.05 (s, 1H) , 6.40 (s, 1H) , 4.14 (d, J = 6.0 Hz, 1H) , 3.97 (d, J = 8.5 Hz, 1H) , 3.86 (d, J = 15.4 Hz, 4H) , 3.76 (d, J = 11.2 Hz, 1H) , 3.64 (d, J = 6.5 Hz, 4H) , 3.51 (t, J = 11.0 Hz, 1H) , 3.08 (t, J = 10.7 Hz, 1H) , 2.28 (s, 3H) , 1.15 (d, J = 6.6 Hz, 3H) .

47b:

¹H NMR (400 MHz, DMSO-d6) δ 8.49 (s, 1H) , 7.93 (s, 1H) , 7.04 (s, 1H) , 6.31 (s, 1H) , 3.98 (d, J = 8.3 Hz, 2H) , 3.93 –3.85 (m, 4H) , 3.73 (d, J = 11.1 Hz, 1H) , 3.66 (d, J = 11.8 Hz, 4H) , 3.50 (t, J = 10.4 Hz, 1H) , 3.05 (dd, J = 12.8, 9.1 Hz, 1H) , 2.27 (s, 3H) , 0.91 (d, J = 6.6 Hz, 3H) .

Example 48

Synthesis of 4, 4-difluoro-1- [3- (3-methyl-1H-pyrazol-5-yl) -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl] cyclohexan-1-ol

Step 1. (3R) -4- [7- (4, 4-difluorocyclohex-1-en-1-yl) -3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] -3-methylmorpholine

To a solution of (3R) -4- {7-chloro-3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine (150 mg, 0.35 mmol) in dioxane (3 mL) were added 2- (4, 4-difluorocyclohex-1-en-1-yl) -4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolane (169 mg, 0.69 mmol) , Pd (PPh₃) ₄ (40 mg, 0.04 mmol) and K₂CO₃ (48 mg, 0.35 mmol) . The reaction was stirred at 100 ℃ overnight under nitrogen atmosphere. LC-MS showed the reaction was completed. The reaction was diluted with EA (50 mL) and water (50 mL) . The organic phase was washed with brine (50 mL) , dried over anhydrous Na₂SO₄, filtered and concentrated in vacuo. The residue was purified by flash column chromatography (Silica gel, 0～100%ethyl acetate in petroleum ether) to give (3R) -4- [7- (4, 4-difluorocyclohex-1-en-1-yl) -3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] -3-methylmorpholine (150 mg, yield: 84%) as a yellow solid. LC/MS (ESI) : m/z 516 (M+H) ⁺.

Step 2. 4, 4-difluoro-1- {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} cyclohexan-1-ol

To a suspension of (3R) -4- [7- (4, 4-difluorocyclohex-1-en-1-yl) -3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] -3-methylmorpholine (150 mg, 0.29 mmol) in Isopropyl alcohol (1 mL) and DCM (5 mL) were added phenylsilane (0.07 mL, 0.58 mmol) and Mn (TMHD) ₃ (18 mg, 0.03 mmol) . After completion of the addition, the mixture was stirred at room temperature overnight. LC-MS showed the reaction was completed. The organic phase was washed with brine (50 mL) , dried over anhydrous Na₂SO₄, filtered and concentrated in vacuo. The residue was purified by flash column chromatography (Silica gel, 0～100%ethyl acetate in petroleum ether) to give to give 4, 4-difluoro-1- {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} cyclohexan-1-ol (70 mg, yield: 45%) as a yellow solid. LC/MS (ESI) : m/z 534 (M+H) ⁺.

Step 3. 4, 4-difluoro-1- [3- (3-methyl-1H-pyrazol-5-yl) -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl] cyclohexan-1-ol

To a solution of 4, 4-difluoro-1- {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -5- [(3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} cyclohexan-1-ol (70 mg, 0.13 mmol) in DCM (3 mL) was added HCl/dioxane (3 mL) . The mixture was stirred at room temperature for 1 hour. LC-MS showed the reaction was completed. The reaction was concentrated in vacuo to give crude product, which was purified by prep-HPLC (C18, 10-95%, MeCN in H₂O with 0.1%CHOOH) to afford the 4, 4-difluoro-1- [3- (3-methyl-1H-pyrazol-5-yl) -5- [ (3R) -3-methylmorpholin-4-yl] -[1, 2] thiazolo [4, 5-b] pyridin-7-yl] cyclohexan-1-ol (12 mg, yield: 20%) . LC/MS (ESI) : m/z 450 (M+H) ⁺.

¹H NMR (400 MHz, DMSO-d6) δ 13.09 (s, 1H) , 7.08 (d, J = 12.5 Hz, 2H) , 6.30 (s, 1H) , 4.58 (d, J = 6.4 Hz, 1H) , 4.12 (d, J = 12.0 Hz, 1H) , 4.01 (d, J = 8.1 Hz, 1H) , 3.79 (d, J = 11.4 Hz, 1H) , 3.70 (d, J = 8.6 Hz, 1H) , 3.55 (t, J = 10.4 Hz, 1H) , 3.26 –3.13 (m, 1H) , 2.39 –2.09 (m, 7H) , 1.98 (dd, J = 55.2, 16.6 Hz, 4H) , 1.20 (d, J = 6.6 Hz, 3H) .

Example 49

Synthesis of 1-cyclopropyl-4- [3- (3-methyl-1H-pyrazol-5-yl) -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl] piperidin-4-ol

Step 1. (3R) -3-methyl-4- (3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) -7- (1, 2, 3, 6-tetrahydropyridin-4-yl) isothiazolo [4, 5-b] pyridin-5-yl) morpholine

To a solution of (3R) -4- {7-chloro-3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine (250 mg, 0.58 mmol) and 4- (tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1, 2, 3, 6-tetrahydropyridine (144 mg, 0.69 mmol) in dioxane (2 mL) and H₂O (0.5 mL) were added Pd (PPh₃) ₄ (67 mg, 0.06 mmol) followed by dipotassium carbonate (398 mg, 2.88 mmol) under N₂ with stirring. The mixture was refluxed and stirred for 16 hours until the starting material was consumed completely. The reaction mixture was cooled to room temperature and concentrated in vacuum to remove most of solvent. The residue was poured into water (20 mL) and extracted with EA (20 mL) . The organic layer was dried over Na₂SO₄, filtered and concentrated. The residue was purified by flash column chromatography (Silica gel, 0～100%ethyl acetate in petroleum ether) to give to give (3R) -3-methyl-4- {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -7- (1, 2, 3, 6-tetrahydropyridin-4-yl) - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} morpholine (150 mg, yield: 54%) as a yellow solid.

Step 2. (3R) -4- (7- (1-cyclopropyl-1, 2, 3, 6-tetrahydropyridin-4-yl) -3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine

To a solution of (3R) -3-methyl-4- {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -7- (1, 2, 3, 6-tetrahydropyridin-4-yl) - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} morpholine (150 mg, 0.31 mmol) and cyclopropylboronic acid (80 mg, 0.94 mmol) in DCE (3 mL) were added disodium carbonate (165 mg, 1.56 mmol) followed by Py (71 mg, 0.62 mmol) and Cu (OAc) ₂ (6 mg, 0.03 mmol) under N₂ with stirring. The mixture was refluxed and stirred at 70℃ for 16 hours until the starting material was consumed completely. The reaction mixture was cooled to room temperature and concentrated in vacuum to remove most of solvent. The residue was poured into water (20 mL) and extracted with EA (20 mL) . The organic layer was dried over Na₂SO₄, filtered and concentrated. The residue was purified by flash column chromatography (Silica gel, 0～100%ethyl acetate in petroleum ether) to give to give (3R) -4- (7- (1-cyclopropyl-1, 2, 3, 6-tetrahydropyridin-4-yl) -3- (3-methyl-1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-5-yl) isothiazolo [4, 5-b] pyridin-5-yl) -3-methylmorpholine (90 mg, yield: 56%) as a yellow oil.

Step 3. 1-cyclopropyl-4- {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} piperidin-4-ol

To a suspension of (3R) -4- [7- (1-cyclopropyl-1, 2, 3, 6-tetrahydropyridin-4-yl) -3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] -3-methylmorpholine (90 mg, 0.17 mmol) in isopropyl alcohol (0.5 mL) and DCM (2.5 mL) were added phenylsilane (0.02 mL, 0.17 mmol) and Mn (TMHD) ₃ (104 mg, 0.17 mmol) . After completion of the addition , the mixture was stirred at room temperature overnight. LC-MS showed the reaction was completed. The reaction was diluted with EA (30 mL) and water (30 mL) . The organic phase was washed with brine (30 mL) , dried over anhydrous Na₂SO₄, filtered and concentrated in vacuo to give crude product. The residue was purified by flash column chromatography (Silica gel, 0～100%ethyl acetate in petroleum ether) to give 1-cyclopropyl-4- {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} piperidin-4-ol (50 mg, 54%) as a yellow solid. LC/MS (ESI) : m/z 539 (M+H) ⁺.

Step 4. 1-cyclopropyl-4- [3- (3-methyl-1H-pyrazol-5-yl) -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl] piperidin-4-ol

To a solution of 1-cyclopropyl-4- {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} piperidin-4-ol (50 mg, 0.09 mmol) in DCM (2 mL) was added HCl/dioxane (2 mL) . The mixture was stirred at room temperature for1 hour. LC-MS showed the reaction was completed. The reaction was concentrated in vacuo to give a crude product, which was purified by prep-HPLC (C18, 10-95%, MeCN in H₂O with 0.1%HCOOH) to afford the 1-cyclopropyl-4- [3- (3-methyl-1H-pyrazol-5-yl) -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl] piperidin-4-ol (6 mg, yield: 14%) . LC/MS (ESI) : m/z 455 (M+H) ⁺.

¹H NMR (400 MHz, CDCl₃) δ 7.07 (s, 1H) , 7.00 (s, 1H) , 5.94 (s, 1H) , 4.53 (s, 1H) , 4.11 (d, J = 12.6 Hz, 1H) , 3.99 (d, J = 8.5 Hz, 1H) , 3.77 (d, J = 11.3 Hz, 1H) , 3.68 (d, J = 9.0 Hz, 1H) , 3.53 (t, J = 10.2 Hz, 1H) , 3.18 (t, J = 11.0 Hz, 1H) , 2.73 (d, J = 57.7 Hz, 4H) , 2.28 (s, 4H) , 2.02 (s, 2H) , 1.71 (s, 3H) , 1.18 (d, J = 6.6 Hz, 3H) , 0.38 (d, J = 57.6 Hz, 4H) .

Example 50

Synthesis of 1-methyl-4- [3- (3-methyl-1H-pyrazol-5-yl) -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl] piperidin-4-ol

Step 1. (3R) -3-methyl-4- [7- (1-methyl-1, 2, 3, 6-tetrahydropyridin-4-yl) -3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] morpholine

To a solution of (3R) -4- {7-chloro-3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine (100 mg, 0.23 mmol) in dioxane (2 mL) were added 1-methyl-4- (tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1, 2, 3, 6-tetrahydropyridine (103 mg, 0.46 mmol) , Pd (PPh₃) ₄ (27 mg, 0.02 mmol) and K₂CO₃ (95 mg, 0.69 mmol) . The reaction was stirred at 100℃ overnight under nitrogen atmosphere. LC-MS showed the reaction was completed. The reaction was diluted with EA (50 mL) and water (50 mL) . The organic phase was washed with brine (50 mL) , dried over anhydrous Na₂SO₄, filtered and concentrated in vacuo. The residue was purified by flash column chromatography (Silica gel, 0～100%ethyl acetate in petroleum ether) to give (3R) -3-methyl-4- [7- (1-methyl-1, 2, 3, 6-tetrahydropyridin-4-yl) -3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] morpholine (110 mg, yield: 96%) as a yellow solid. LC/MS (ESI) : m/z 495 (M+H) ⁺.

Step 2. 1-methyl-4- {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} piperidin-4-ol

To a suspension of (3R) -3-methyl-4- [7- (1-methyl-1, 2, 3, 6-tetrahydropyridin-4-yl) -3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] morpholine (110 mg, 0.22 mmol) in Isopropyl alcohol (1 mL) and DCM (5 mL) were added phenylsilane (0.03 mL, 0.22 mmol) and Mn (TMHD) ₃ (134 mg, 0.22 mmol) . After completion of the addition, the mixture was stirred at room temperature overnight. LC-MS showed the reaction was completed. The reaction was diluted with EA (50 mL) and water (50 mL) . The organic phase was washed with brine (50 mL) , dried over anhydrous Na₂SO₄, filtered and concentrated in vacuo to give crude product. The residue was purified by flash column chromatography (Silica gel, 0～100%ethyl acetate in petroleum ether) to give to give 1-methyl-4- {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} piperidin-4-ol (80 mg, yield: 70%) as a yellow solid. LC/MS (ESI) : m/z 513 (M+H) ⁺.

Step 3. 1-methyl-4- [3- (3-methyl-1H-pyrazol-5-yl) -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl] piperidin-4-ol

To a solution of 1-methyl-4- {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} piperidin-4-ol (80 mg, 0.16 mmol) in DCM (4 mL) was added HCl/dioxane (4 mL) . The mixture was stirred at room temperature for1 hour. LC-MS showed the reaction was completed. The reaction was concentrated in vacuo to give crude product, which was purified by prep-HPLC (C18, 10-95%, MeCN in H₂O with 0.1%CHOOH) to afford the 1-methyl-4- [3- (3-methyl-1H-pyrazol-5-yl) -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl] piperidin-4-ol (22 mg, yield: 33%) . LC/MS (ESI) : m/z 429 (M+H) ⁺.

¹H NMR (400 MHz, DMSO-d6) δ 7.07 (d, J = 15.5 Hz, 2H) , 5.97 (s, 1H) , 4.54 (d, J = 6.5 Hz, 1H) , 4.12 (d, J = 11.8 Hz, 1H) , 4.01 (dd, J = 11.2, 3.0 Hz, 1H) , 3.79 (d, J = 11.3 Hz, 1H) , 3.70 (dd, J = 11.3, 2.6 Hz, 1H) , 3.60 –3.50 (m, 2H) , 3.28 –3.09 (m, 2H) , 2.72 (d, J = 10.7 Hz, 1H) , 2.46 (d, J = 11.1 Hz, 1H) , 2.29 (d, J = 3.4 Hz, 6H) , 2.16 (dt, J = 13.1, 9.1 Hz, 2H) , 1.74 (d, J = 13.1 Hz, 2H) , 1.20 (d, J = 6.6 Hz, 3H) .

Example 51

Synthesis of (3R) -4- [7- (4, 4-difluoropiperidin-1-yl) -3- (3-methyl-1H-pyrazol-5-yl) - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] -3-methylmorpholine

Step 1. (3R) -4- [7- (4, 4-difluoropiperidin-1-yl) -3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] -3-methylmorpholine

To a solution of (3R) -4- {7-chloro-3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine (100 mg, 0.23 mmol) in DMF (2 mL) was added 4, 4-difluoro-1- (tetramethyl-1, 3, 2-dioxaborolan-2-yl) piperidine (114 mg, 0.46 mmol) , RuPhos (22 mg, 0.05 mmol) , RuPhos Pd G3 (19 mg, 0.02 mmol) and Cs₂CO₃ (150 mg, 0.46 mmol) . The reaction was stirred at 100℃overnight under nitrogen atmosphere. LC-MS showed the reaction was completed. The reaction was diluted with EA (50 mL) and water (50 mL) . The organic phase was washed with brine (50 mL) , dried over anhydrous Na₂SO₄, filtered and concentrated in vacuo. The residue was purified by flash column chromatography (Silica gel, 0～100%ethyl acetate in petroleum ether) to give (3R) -4- [7- (4, 4-difluoropiperidin-1-yl) -3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] -3-methylmorpholine (100 mg, yield: 84%) as a yellow solid. LC/MS (ESI) : m/z 519 (M+H) ⁺.

Step 2. (3R) -4- [7- (4, 4-difluoropiperidin-1-yl) -3- (3-methyl-1H-pyrazol-5-yl) - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] -3-methylmorpholine

To a solution of (3R) -4- [7- (4, 4-difluoropiperidin-1-yl) -3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] -3-methylmorpholine (100 mg, 0.19 mmol) in DCM (3 mL) was added HCl/dioxane (3 mL) and stirred at room temperature for 1 hour. LC-MS showed the reaction was completed. The reaction was concentrated in vacuo to give a crude product, which was purified by prep-HPLC (C18, 10-95%, MeCN in H₂O with 0.1%CHOOH) to afford the (3R) -4- [7- (4, 4-difluoropiperidin-1-yl) -3- (3-methyl-1H-pyrazol-5-yl) - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] -3-methylmorpholine (56 mg, yield: 67%) . LC/MS (ESI) : m/z 435 (M+H) ⁺.

¹H NMR (400 MHz, DMSO-d6) δ 7.09 (s, 1H) , 6.43 (s, 1H) , 4.50 (d, J = 5.7 Hz, 1H) , 4.03 (t, J = 12.6 Hz, 2H) , 3.79 (d, J = 11.2 Hz, 1H) , 3.70 (d, J = 10.9 Hz, 1H) , 3.65 –3.58 (m, 4H) , 3.57 –3.51 (m, 1H) , 3.22 (dd, J = 17.5, 7.9 Hz, 1H) , 2.30 (s, 3H) , 2.23 –2.09 (m, 4H) , 1.21 (d, J = 6.6 Hz, 3H) .

Example 52

Synthesis of (R) -2- (3- (3-methyl-1H-pyrazol-5-yl) -5- (3-methylmorpholino) isothiazolo [4, 5-b] pyridin-7-yl) propan-2-ol

Step 1. (3-methyl-1H-pyrazol-5-yl) boronic acid

To a solution of [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] boronic acid (500 mg, 2.38 mmol) in DCM (5 mL) was added HCl/dioane (7 mL) and stirred at 60℃ overnight. LC-MS showed the reaction was completed. The reaction was concentrated in vacuo to give crude product (3-methyl-1H-pyrazol-5-yl) boronic acid (290 mg, yield: 97%) as a white solid. LC/MS (ESI) : m/z 127 (M+H) ⁺.

Step 2. (3R) -4- [7-chloro-3- (3-methyl-1H-pyrazol-5-yl) - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] -3-methylmorpholine

To a solution of 7-chloro-5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-3-yl trifluoromethanesulfonate (800 mg, 1.91 mmol) in dioxane (10 mL) were added (3-methyl-1H-pyrazol-5-yl) boronic acid (289 mg, 2.30 mmol) , Pd (PPh₃) ₄ (221 mg, 0.19 mmol) and K₂CO₃ (794 mg, 5.74 mmol) . The reaction was stirred at 100℃ overnight under nitrogen atmosphere. LC-MS showed the reaction was completed. The reaction was diluted with EA (50 mL) and water (50 mL) . The organic phase was washed with brine (50 mL) , dried over anhydrous Na₂SO₄, filtered and concentrated in vacuo. The residue was purified by flash column chromatography (Silica gel, 0～100%ethyl acetate in petroleum ether) to give (3R) -4- [7-chloro-3- (3-methyl-1H-pyrazol-5-yl) - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] -3-methylmorpholine (200 mg, yield: 30%) as a yellow solid. LC/MS (ESI) : m/z 350 (M+H) ⁺.

Step 3. (3R) -3-methyl-4- [3- (3-methyl-1H-pyrazol-5-yl) -7- [2- (oxan-2-yl) -2H-1, 2, 3-triazol-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] morpholine

To a solution of (3R) -4- [7-chloro-3- (3-methyl-1H-pyrazol-5-yl) - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] -3-methylmorpholine (200 mg, 0.57 mmol) in dioxane (3 mL) were added 2- (oxan-2-yl) -4- (tetramethyl-1, 3, 2-dioxaborolan-2-yl) -2H-1, 2, 3-triazole (319 mg, 1.14 mmol) , Pd (PPh₃) ₄ (66 mg, 0.06 mmol) and K₂CO₃ (237 mg, 1.71 mmol) . The reaction was stirred at 100℃ overnight under nitrogen atmosphere. LC-MS showed the reaction was completed. The reaction was diluted with EA (50 mL) and water (50 mL) . The organic phase was washed with brine (50 mL) , dried over anhydrous Na₂SO₄, filtered and concentrated in vacuo. The residue was purified by flash column chromatography (Silica gel, 0～100%ethyl acetate in petroleum ether) to give (3R) -3-methyl-4- [3- (3-methyl-1H-pyrazol-5-yl) -7- [2- (oxan-2-yl) -2H-1, 2, 3-triazol-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] morpholine (100 mg, yield: 37%) as a yellow solid. LC/MS (ESI) : m/z 467 (M+H) ⁺.

Step 4. (3R) -3-methyl-4- [3- (3-methyl-1- { [2- (trimethylsilyl) ethoxy] methyl} -1H-pyrazol-5-yl) -7- [2- (oxan-2-yl) -2H-1, 2, 3-triazol-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] morpholine

To a solution of (3R) -3-methyl-4- [3- (3-methyl-1H-pyrazol-5-yl) -7- [2- (oxan-2-yl) -2H-1, 2, 3-triazol-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] morpholine (350 mg, 0.75 mmol) in THF (5 mL) were added NaH (45 mg, 1.50 mmol) and SEMCl (0.2 mL, 0.90 mmol) . The mixture was stirred at 0℃ ～ room temperature for 2 h. TLC showed the reaction was completed. The mixture was added H₂O (10 mL) and extracted with EA (10 mL x 3) . The combined organic layer was washed with brine (10 mL) , dried over Na₂SO₄ and evaporated to dryness. The residue was purified by flash column chromatography on silica gel to give (3R) -3-methyl-4- [3- (3-methyl-1- { [2- (trimethylsilyl) ethoxy] methyl} -1H-pyrazol-5-yl) -7- [2- (oxan-2-yl) -2H-1, 2, 3- triazol-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] morpholine (230 mg, yield: 51%) as a colorless oil. LC/MS (ESI) : m/z 598.1 [M+H] ⁺.

Step 5. (3R) -3-methyl-4- [3- (3-methyl-1- { [2- (trimethylsilyl) ethoxy] methyl} -1H-pyrazol-5-yl) -7- (2H-1, 2, 3-triazol-4-yl) - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] morpholine

To a solution of (3R) -3-methyl-4- [3- (3-methyl-1- { [2- (trimethylsilyl) ethoxy] methyl} -1H-pyrazol-5-yl) -7- [2- (oxan-2-yl) -2H-1, 2, 3-triazol-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] morpholine (350 mg, 0.59 mmol) in DCM (5 mL) was added TFA (3 mL) . The reaction was stirred at room temperature for 2 h. TLC showed the reaction was completed. After removing the solvent, the reaction was purified by flash column chromatography on silica get to give (3R) -3-methyl-4- [3- (3-methyl-1- { [2- (trimethylsilyl) ethoxy] methyl} -1H-pyrazol-5-yl) -7- (2H-1, 2, 3-triazol-4-yl) - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] morpholine (220 mg, yield: 73%) as a colorless oil. LC/MS (ESI) : m/z 513.1 [M+H] ⁺.

Step 6. (3R) -4- {7- [1- (difluoromethyl) -1H-1, 2, 3-triazol-4-yl] -3- (3-methyl-1- { [2- (trimethylsilyl) ethoxy] methyl} -1H-pyrazol-5-yl) - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine

To a solution of (3R) -3-methyl-4- [3- (3-methyl-1- { [2- (trimethylsilyl) ethoxy] methyl} -1H-pyrazol-5-yl) -7- (2H-1, 2, 3-triazol-4-yl) - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] morpholine (230 mg, 0.45 mmol) in THF (5 mL) were added diethyl (bromodifluoromethyl) phosphonate (240 mg, 0.90 mmol) and NaH (36 mg, 0.90 mmol) at 0℃. The reaction stirred at 0℃ for 2 h. TLC showed the reaction was completed. The mixture was added H₂O (10 mL) and extracted with EA (10 mL x 3) . The combined organic layer was washed with brine (10 mL) , dried over Na₂SO₄ and evaporated to dryness. The residue was purified by flash column chromatography on silica gel to give (3R) -3-methyl-4- [3- (3-methyl-1H-pyrazol-5-yl) -7- (2H-1, 2, 3-triazol-4-yl) - [1, 2] thiazolo [4, 5-b] pyridin-5-yl] morpholine (50 mg, yield: 29%) as a colorless oil. LC/MS (ESI) : m/z 563.1 [M+H] ⁺.

Step 7. (3R) -4- {7- [1- (difluoromethyl) -1H-1, 2, 3-triazol-4-yl] -3- (3-methyl-1H-pyrazol-5-yl) - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine

To a solution of (3R) -4- {7- [1- (difluoromethyl) -1H-1, 2, 3-triazol-4-yl] -3- (3-methyl-1- { [2- (trimethylsilyl) ethoxy] methyl} -1H-pyrazol-5-yl) - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine (50 mg, 0.09 mmol) in DCM (5 mL) was added TFA (2 mL) stirred at room temperature for 2 h. LCMS showed the reaction was completed. The reaction mixture was concentrated under reduced pressure. The residue was purified by Prep-HPLC (C18, 10-95%, MeOH in H2O with 0.1% HCOOH) to give (3R) -4- {7- [1- (difluoromethyl) -1H-1, 2, 3-triazol-4-yl] -3- (3-methyl-1H-pyrazol-5-yl) - [1, 2] thiazolo [4, 5-b] pyridin-5-yl} -3-methylmorpholine (5 mg, yield: 13 %) . LC/MS (ESI) : m/z 433.1 [M+H] ⁺.

¹H NMR (400 MHz, DMSO-d6) δ 9.80 (s, 1H) , 8.39 (dd, J = 118.8, 60.5 Hz, 1H) , 7.81 (s, 1H) , 7.14 (s, 1H) , 4.58 (d, J = 6.0 Hz, 1H) , 4.18 (d, J = 12.9 Hz, 1H) , 4.08 (d, J = 8.8 Hz, 1H) , 3.86 (d, J = 11.5 Hz, 1H) , 3.76 (d, J = 9.0 Hz, 1H) , 3.60 (t, J = 10.9 Hz, 1H) , 3.18 (s, 1H) , 2.31 (s, 3H) , 1.27 (d, J = 6.5 Hz, 3H) .

Example 53

Synthesis of cyano [methyl ( {1- [3- (3-methyl-1H-pyrazol-5-yl) -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl] cyclopropyl} ) oxo-λ^6-sulfanylidene] amine

Step 1. (cyanoimino) (methyl) ( {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} methyl) -λ^6-sulfanone

To a solution of imino (methyl) ( {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} methyl) -λ^6-sulfanone (80 mg, 0.16 mmol) in DCM (5 mL) was added Py (0.03 mL, 0.33 mmol) and BrCN (0.024 mL, 0.33 mmol) at -78℃, and stirred at -78℃-room temperature for 2 hours. LCMS showed the reaction was completed. The reaction mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography (Silica gel, 0～100%ethyl acetate in petroleum ether) to give the desired product (cyanoimino) (methyl) ( {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -5- [(3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} methyl) -λ^6-sulfanone (40 mg, yield: 48%) as a white solid. LC/MS (ESI) : m/z 516.7 [M+H] ⁺.

Step 2. cyano [methyl (1- {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} cyclopropyl) oxo-λ^6-sulfanylidene] amine

To a solution of (cyanoimino) (methyl) ( {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} methyl) -λ^6-sulfanone (40 mg, 0.08 mmol) in toluene (3 mL) was added 1, 2-dibromoethane (29 mg, 0.16 mmol) , TBAB (13 mg, 0.04 mmol) and NaOH (31 mg, 0.78 mmol) . The reaction was stirred at 60℃ for 2 hours. LC-MS showed the reaction was completed. The reaction was diluted with EA (50 mL) and water (50 mL) . The organic phase was washed with brine (50 mL) , dried over anhydrous Na₂SO₄, filtered and concentrated in vacuo. The residue was purified by flash column chromatography (Silica gel, 0～100%ethyl acetate in petroleum ether) to give cyano [methyl (1- {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} cyclopropyl) oxo-λ^6-sulfanylidene] amine (15 mg, yield: 36%) as a yellow solid. LC/MS (ESI) : m/z 542 (M+H) ⁺.

Step 3. cyano [methyl ( {1- [3- (3-methyl-1H-pyrazol-5-yl) -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl] cyclopropyl} ) oxo-λ^6-sulfanylidene] amine

To a solution of cyano [methyl (1- {3- [3-methyl-1- (oxan-2-yl) -1H-pyrazol-5-yl] -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl} cyclopropyl) oxo-λ^6-sulfanylidene] amine (15 mg, 0.03 mmol) in DCM (2 mL) was added HCl/dioxane (2 mL) . The mixture was stirred at room temperature for 1 hour. LC-MS showed the reaction was completed. The reaction was concentrated in vacuo to give a crude product, which was purified by prep-HPLC (C18, 10-95%, MeCN in H₂O with 0.1%CHOOH) to afford cyano [methyl ( {1- [3- (3-methyl-1H-pyrazol-5-yl) -5- [ (3R) -3-methylmorpholin-4-yl] - [1, 2] thiazolo [4, 5-b] pyridin-7-yl] cyclopropyl} ) oxo-λ^6-sulfanylidene] amine (2 mg, yield: 15%) . LC/MS (ESI) : m/z 458(M+H) ⁺.

¹H NMR (400 MHz, DMSO-d6) δ 7.59 (d, J = 3.8 Hz, 1H) , 7.11 (s, 1H) , 4.50 (s, 1H) , 4.15 (d, J = 13.9 Hz, 1H) , 4.06 (d, J = 11.2 Hz, 1H) , 3.83 (d, J = 11.5 Hz, 1H) , 3.72 (dd, J = 11.4, 2.8 Hz, 1H) , 3.58 (t, J = 9.6 Hz, 4H) , 3.25 (dd, J = 12.7, 3.6 Hz, 1H) , 2.31 (s, 3H) , 2.05 –1.91 (m, 2H) , 1.79 (td, J = 11.9, 6.2 Hz, 2H) , 1.26 (dd, J =6.6, 3.0 Hz, 5H) .

Biochemical Assays

Assay 1: ATR inhibition assay

Detection of ATR kinase activity utilized the Mobility shift assay to measure the phosphorylation of the substrate protein FAM-RAD17 (GL, Cat. No. 514318, Lot. No. P19042-MJ524315) . The assay was developed and conducted at Chempartner. All the test compounds were dissolved in 100%DMSO at concentration of 20 mM, then prepare compounds and conducted the assay as follows:

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

2) Serially dilute the compound by transferring 20μl to 60μl of 100%DMSO in the next well and so forth for a total of 10 concentrations.

3) Add 100 μl of 100%DMSO to two empty wells for no compound control and no enzyme control in the same 96-well plate. Mark the plate as source plate.

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

5) Transfer 60 nl compounds to assay plate by Echo.

6) Add ATR kinase (Eurofins, Cat. No. 14-953, Lot. No. D14JP007N) into Kinase base buffer (50 mM HEPES, pH 7.5; 0.0015%Brij-35; 0.01%Triton) to prepare 2 x enzyme solution, then add 10 μl of 2x enzyme solution to each well of the 384-well assay plate, incubate at room temperature for 10 min.

7) Add FAM-RAD17 and ATP (Sigma, Cat. No. A7699-1G, CAS No. 987-65-5) in the kinase base buffer to prepare 2x peptide solution, then add 10μl to the assay plate.

8) Incubate at 28℃ for specified period of time. 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 reaction.

9) Collect data on Caliper. Convert conversion values to inhibition values.

Percent inhibition = (max-conversion) / (max-min) *100

wherein “max” stands for DMSO control; “min” stands for low control.

Fit the data in XLFit excel add-in version 5.4.0.8 to obtain IC50 values. Equation used is:

Y=Bottom + (Top-Bottom) / (1+ (IC50/X) ^HillSlope)

wherein X means concentration in a format not transformed to logarithms.

The following Table 2 lists the IC₅₀ values for exemplary compounds of Formula (I) .

Table 2

For the other compounds provided herein for which the results are not shown, all have an IC₅₀ against ATR kinase of no more than 1000 nM. Some of these compounds have an IC₅₀ against ATR kinase of no more than 500 nM, some no more than 400 nM, some no more than 300 nM, some no more than 200 nM, or no more than 100 nM, or even no more than 50 nM.

Therefore, as determined by ATR inhibition assay, the compounds of the present disclosure have a good inhibitory effect on ATR kinase activity.

Assay 2: Tumor Cell Anti-proliferation Assay (CTG Assay)

Human colorectal cancer cells HT-29 (HTB-38) and LoVo (CCL-229) were selected for the CTG assay, the two cell lines were originally obtained from the American Type Culture Collection (ATCC) . FBS and appropriate additives were added into base medium to prepare complete medium, then the cell layer was briefly rinsed with 0.25% (w/v) Trypsin-0.038% (w/v) EDTA solution to remove all traces of serum that contains trypsin inhibitor. After that, appropriate volume of Trypsin-EDTA solution was added to flask and cells under an inverted microscope were observed until cell layer was dispersed. At last, appropriate volume of complete growth medium was added and cells were aspirated by gently pipetting. Numbers were collected and counted with Vi-cell XR and cell density was adjusted, cells were seeded into 96-well opaque-walled clear bottom tissue-culture treated plates in the CO₂ incubator for 20-24 hours. All the test compounds were at 10 mM in DMSO. Compounds were then added to the cell media in 3-fold serial dilutions, the final DMSO concentration was 0.5%. Plates were incubated for 96h at 5%CO₂, 37℃. Before the measurement, the appropriate volume of CellTiter-Glo Buffer was transferred into the amber bottle containing CellTiter-Glo substrate to reconstitute the lyophilized enzyme/substrate mixture, mixed gently, thereby forming the CellTiter-Glo Reagent (Promega Cat. No. G7573) . The plate and its contents were equilibrated to room temperature for approximately 30 minutes, then 100 μL of CellTiter-Glo Reagent was added to the assay plate, the contents were mixed for 2 minutes on an orbital shaker to induce cell lysis, incubated at room temperature for 10 minutes to stabilize luminescent signal. At last the clear bottom was pasted with white back seal and luminescence was recorded with Enspire. IC₅₀ and GI₅₀ values were calculated with XLFit curve fitting software using 4 Parameter Logistic Model Y=Bottom + (Top-Bottom) / (1+ (IC50/X) ^HillSlope) .

The following Table 3 provides the IC₅₀ values for exemplary compounds of Formula (I) .

Table 3

Pharmacokinetics Study

The purpose of this study is to determine the pharmacokinetics parameters in plasma of compounds in fasted male CD-1 mice following intravenous (IV) or oral (PO) administration.

Test Article Preparation

The formulations for intravenous and oral administrations were:

For IV adminstration: 0.20 mg/mL test compound in vehicle (10%DMSO+10%Solutol+80%Water)

For PO administration: 1 mg/mL test compound in vehicle (10%DMSO+10%Solutol+80%Water)

Study Design

*The animals were fasted prior to oral administration. Food supply to the animals dosed orally were resumed 4 hours post-dose.

Administration

The test article was be administered via a single IV or PO dosing.

Collection Intervals

IV group: Post-dose at 5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 8 h and 24 h.

PO group: Post-dose at 15 min, 30 min, 1 h, 2 h, 4 h, 6 h, 8 h and 24 h.

Analysis Procedure

The PK blood samples were centrifuged at approximately 6800G for 6 minutes at 2-8℃ and the resulting plasma was transferred to appropriately labeled tubes within 2 hour of blood collection/centrifugation and stored frozen at approximately -70℃.

Method development and biological samples analysis for the test articles (Sodium heparin anticoagulant) were be performed by means of LC-MS/MS. The analytical results were be confirmed using quality control samples for intra-assay variation. The accuracy of >66.7%of the quality control samples were be between 80 -120%of the known value (s) .

Pharmacokinetics Analysis

Standard set of parameters including Area Under the Curve (AUC _(0-t) and AUC_(0-∞) ) , elimination half-live (T_1/2) , maximum plasma concentration (Cmax) and time to reach maximum plasma concentration (Tmax) and other parameters were be calculated using Phoenix WinNonlin 7.0 (Pharsight, USA) by the Study Director.

The following Table 4 provides the results for exemplary compounds of Formula (I) .

The foregoing description is considered as illustrative only of the principles of the present disclosure. Further, since numerous modifications and changes will be readily apparent to those skilled in the art, it is not desired to limit the invention to the exact construction and process shown as described above. Accordingly, all suitable modifications and equivalents may be considered to fall within the scope of the invention as defined by the claims that follow.

Claims

A compound having Formula (I) :

or a pharmaceutically acceptable salt thereof,

wherein

R¹ and R² each independently is alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl or heteroaryl, wherein said cycloalkyl, heterocyclyl, aryl and heteroaryl are optionally substituted with one or more R^a; or

R¹ and R² together with the sulfur atom to which they are both attached form a heterocyclyl optionally substituted with one or more R^b;

R^a is independently selected from the group consisting of hydroxyl, halogen, cyano, amino, alkyl, alkoxyl, and haloalkyl;

R^b is independently selected from the group consisting of alkyl, alkenyl, alknyl, alkoxy, cyano, halogen, hydroxyl, amino, and nitro, wherein said alkyl, alkenyl and alknyl are optionally substituted with one or more groups independently selected from hydroxyl, halogen, cyano, amino and alkoxyl;

R³ is

Ring A is 5-to 6-membered heterocyclyl or 5-to 6-membered heteroaryl;

R⁴, in each occurrence, is halogen, alkyl, haloalkyl, or cycloalkyl; and

m is 0, 1, 2 or 3.
The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R¹ and R² are each independently C_1-3 alkyl, C_3-6 cycloalkyl or C_5-12 aryl.
The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R¹ and R² are independently C_1-3 alkyl.
The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R¹ and R² are independently C_3-6 cycloalkyl.
The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein one of R¹ and R² is C_1-3 alkyl, and the other is C_3-6 cycloalkyl or C_5-12 aryl.
The compound of claim 1, or a pharmaceutically acceptable salt thereof, whereinis selected from the group consisting of:
The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R¹ and R² together with the sulfur atom to which they are both attached form a heterocyclyl optionally substituted with one or more R^b.
The compound of claim 1, or a pharmaceutically acceptable salt thereof, whereinis selected fromeach of which is optionally substituted with one or more R^b.
The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein Ring A is 5-to 6-membered heteroaryl.
The compound of claim 9, or a pharmaceutically acceptable salt thereof, wherein Ring A is pyrazolyl, pyrrolyl, or pyridyl.
The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R⁴ is halogen.
The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R⁴ is alkyl.
The compound of claim 12, or a pharmaceutically acceptable salt thereof, wherein R⁴ is C_1-3 alkyl.
The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R⁴ is haloalkyl.
The compound of claim 13, or a pharmaceutically acceptable salt thereof, wherein R⁴ is C_1-3 haloalkyl.
The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R⁴ is cycloalkyl.
The compound of claim 16, or a pharmaceutically acceptable salt thereof, wherein R⁴ is C_3-6 cycloalkyl.
The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein m is 0, 1 or 2.
The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein is selected from the group consisting of:
The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R³ is
The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R³ is
The compound of any one of preceding claims, having a formula (Ia) :

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

or a pharmaceutically acceptable salt thereof.
A compound selected from the group consisting of:

or a pharmaceutically acceptable salt thereof.
A pharmaceutical composition comprising the compound of any one of claims 1-24 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.
A method for treating cancer, comprising administering an effective amount of a compound of any one of claims 1-24 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of claim 25 to a subject in need thereof.
Use of the compound of any one of claims 1-24 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of claim 23 in the manufacture of a medicament for treating cancer.
A compound of any one of claims 1-24 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of claim 25, for use in the treatment of cancer.
A method for inhibiting ATR kinase in a subject in need thereof, comprising administering an effective amount of a compound of any one of claims 1-24 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of claim 25 to the subject.