Compounds for treating or suppressing relapse of acute myeloid leukemia

Abstract

The present invention relates to compounds for treating acute myeloid leukemia or inhibiting the recurrence of acute myeloid leukemia, and for inhibiting the growth of and / or killing leukemia stem cells.

Description

[0001] This application claims the benefit of International Patent Application No. PCT / CN20 / 79464, filed on March 16, 2020, which is incorporated herein by reference in its entirety.

[0002] Background

[0003] Acute myeloid leukemia (hereinafter also referred to as "AML") is a malignant blood disease with a poor prognosis that usually occurs in adults, and its 5-year survival rate is predicted to be 20%. In some cases, it is possible to temporarily reduce the number of AML cells to a level below the detection limit through AML treatment. This situation is called "complete remission". However, even when complete remission is achieved, AML often recurs, and for many patients, recurrent AML leads to death. Therefore, there is an urgent need to develop more durable treatments for AML.

[0004] While conventional chemotherapeutic agents achieve a temporary remission of AML, the recurrence of AML has been a concern. Specifically, the extremely low survival rate in the case of recurrence has been a serious problem of concern. There is a need for new approaches to treat AML and other blood cancers. Summary of the Invention

[0006] The present invention provides compounds that inhibit the growth of leukemia stem cells and / or treat acute myeloid leukemia or inhibit the recurrence of acute myeloid leukemia.

[0007] In some embodiments, the present invention relates to compounds of formula (I):

[0008]

[0009] Wherein

[0010] Each Q, Y, and Z is independently selected from N and C, and X is N or C-R a ;

[0011] Provided that at least one of Q, X, Y, and Z is N, and

[0012] Each dotted bond is independently a single bond or a double bond such that the bicyclic ring formed thereby is a heteroaryl;

[0013] R 1 Is selected from alkyl, alkenyl, alkynyl, amino, acylamino, cycloalkyl, cycloalkylalkyl, heterocyclic group, heterocyclic group alkyl, aryl, arylalkyl, heteroaryl, and heteroarylalkyl;

[0014] Or

[0015] a) W is N, R 2 Is absent, and A 1 And A 2 Are each CH 2 ;

[0016] b) W is C, R 2 is selected from H, halogen, CN, alkyl, alkoxy, hydroxy, acyloxy and amino, and A 1 and A 2 are each independently selected from CH 2 and O; or

[0017] c) W is C, and R 1 and R 2 together form =CH 2 -acylamino, cycloalkyl and heterocyclic group, and A 1 and A 2 are each independently selected from CH 2 and O;

[0018] R a is selected from H, halogen, CN and alkyl;

[0019] R 9 is selected from H, halogen, alkyl, alkoxy, hydroxy, acyloxy and amino;

[0020] R 10 is H or alkoxy;

[0021] R 11 is aryloxy, heteroaryloxy, aralkyl, alkoxycarbonyl, ureido or -C(O)-aryl; and

[0022] R 12 is selected from H, halogen, CN, alkyl, alkoxy, hydroxy and acyloxy;

[0023] provided that the compound of formula (I) is not

[0024] and if W is C, R 2 、R 9 and R 12 are each hydrogen, and R 11 is unsubstituted phenoxy, then

[0025] R 1 is not -NHCH 2 OH, -NHCH 2 COOH, -NHCH 2 CONH 2 、-NHCH 2 CH 2 NH 2 、-NH(CH 2 ) 2 N(Me) 2 、-NHCH 2-pyridyl, -NHCO-pyridyl, -C(O)OEt, -NH-tetrahydropyran, -NH(CH 2 ) 3 imidazolyl, -NHCH 2 pyrrolidinyl, -NH-pyrrolidinonyl,

[0026] wherein R 13 is selected from H, ethyl, isopropyl, tert-butyl, -CH 2 OCH 3 , -CH 2 CH 2 OH and -CH 2 CH 2 OCH 3 ; and if W is N, R 9 and R 12 are each hydrogen, and R 11 is unsubstituted phenoxy, then R 1 is not piperidinyl.

[0027] The present disclosure further relates to a method of co-inhibiting HCK and BCL-2 in cells, which comprises contacting the cells with a compound disclosed herein and a BCL-2 inhibitor.

[0028] The present disclosure further relates to a method of killing cells having an FLT3-ITD mutation, the method comprising contacting the cells with a compound disclosed herein and a BCL-2 inhibitor.

[0029] The present disclosure further relates to a method of treating acute myeloid leukemia, which comprises co-administering a compound disclosed herein and a BCL-2 inhibitor to a subject.

[0030] Detailed Description

[0031] Definition

[0032] Unless otherwise defined, all technical and scientific terms used herein have the meaning commonly understood by one of ordinary skill in the art to which this disclosure pertains. The following references provide one of ordinary skill in the art with a general definition of many of the terms used in this disclosure: Singleton et al., Dictionary of Microbiology and Molecular Biology (2nd ed. 1994); The Cambridge Dictionary of Science and Technology (Walker ed., 1988); The Glossary of Genetics, 5th Ed., R. Rieger et al. (eds.), Springer Verlag (1991); and Hale & Marham, The Harper Collins Dictionary of Biology (1991). Unless otherwise noted, as used herein, the following terms have the meanings given below.

[0033] In this disclosure, terms such as "comprises," "comprising," "contains," and "has" can have the meanings ascribed to them in U.S. patent law and can mean "includes," "including," etc.; "consisting essentially of" or "consists essentially" likewise have the meanings ascribed to them in U.S. patent law, and the term is open-ended (thus allowing for the presence of more than what is recited, so long as the basic or novel characteristics of what is recited are not changed by the presence of more than what is recited), but excluding prior art embodiments.

[0034] Unless explicitly stated or obvious from the context, as used herein, the term "or" shall be understood to be inclusive. Unless explicitly stated or obvious from the context, as used herein, the terms "a," "an," and "the" shall be understood to be singular or plural.

[0035] The term "acyl" is well recognized in the art and refers to a group represented by the general formula hydrocarbyl C(O)- (preferably alkyl C(O)-).

[0036] The term "amido" is well recognized in the art and refers to an amino group substituted with an acyl group and can be represented, for example, by the formula hydrocarbyl C(O)NH-.

[0037] The term "acyloxy" is well recognized in the art and refers to a group represented by the general formula hydrocarbyl C(O)O- (preferably alkyl C(O)O-).

[0038] The term "alkoxy" refers to an alkyl group having an oxygen attached thereto (preferably a lower alkyl group). Representative alkoxy groups include methoxy, ethoxy, propoxy, tert-butoxy, and the like.

[0039] The term "alkoxyalkyl" refers to an alkyl group substituted with an alkoxy group and can be represented by the general formula alkyl-O-alkyl.

[0040] As used herein, the term "alkenyl" refers to an aliphatic hydrocarbon group containing at least one double bond and is intended to include both "unsubstituted alkenyl" and "substituted alkenyl", where the latter refers to an alkenyl moiety having substituents in place of the hydrogen on one or more carbons of the alkenyl group. Such substituents can be present on one or more carbons that are or are not included in one or more double bonds. In addition, such substituents include all those that are contemplated for alkyl groups as discussed below, except where stability is prohibited. For example, an alkenyl group is expected to be substituted with one or more alkyl, carbocyclic, aryl, heterocyclic, or heteroaryl groups.

[0041] An "alkyl" group or "alkane" is a fully saturated straight-chain or branched-chain non-aromatic hydrocarbon. Typically, unless otherwise specified, a straight-chain or branched-chain alkyl group has from 1 to about 20 carbon atoms, preferably from 1 to about 10. Examples of straight-chain and branched-chain alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, pentyl, hexyl, pentyl, and octyl. C 1 -C 6 Straight-chain or branched-chain alkyl groups are also referred to as "lower alkyl" groups.

[0042] In addition, as used throughout the specification, the examples, and the claims, the term "alkyl" (or "lower alkyl") is intended to include both "unsubstituted alkyl" and "substituted alkyl", where the latter refers to an alkyl moiety having substituents replacing hydrogen on one or more carbons of the hydrocarbon backbone. Unless otherwise specified, such substituents can include, for example, halogen elements, hydroxyl groups, carbonyl groups (such as carboxyl, alkoxycarbonyl, formyl, or acyl), thiocarbonyl groups (such as thioesters, thioacetates, or thiocarboxylates), alkoxy groups, phosphoryl groups, phosphates, phosphonates, hypophosphites, amino groups, amido groups, amidines, imines, cyano groups, nitro groups, azide groups, mercapto groups, alkylthio groups, sulfates, sulfonates, sulfamoyl groups, sulfinylamino groups, sulfonyl groups, heterocyclic groups, aralkyl groups, or aromatic or heteroaromatic moieties. Those skilled in the art will understand that the substituted portions on the hydrocarbon chain can themselves be substituted (if appropriate). For example, the substituents of a substituted alkyl can include amino, azide, imino, amido, phosphoryl (including phosphonates and hypophosphites), sulfonyl (including sulfates, sulfinylamino, sulfamoyl, and sulfonates) and silyl groups, as well as substituted and unsubstituted forms of ethers, alkylthio, carbonyl (including ketones, aldehydes, carboxylates, and esters), -CF 3 and the substituted and unsubstituted forms of -CN, etc. Exemplary substituted alkyls are described below. The cycloalkyl can be further substituted with alkyl, alkenyl, alkoxy, alkylthio, aminoalkyl, carbonyl-substituted alkyl, -CF 3 -CN, etc.

[0043] When used in conjunction with chemical moieties such as acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy, the term "C x-y " means a group containing from x to y carbons in the chain. For example, the term "C x-y alkyl" refers to a substituted or unsubstituted saturated hydrocarbon group (including straight-chain alkyl groups and branched-chain alkyl groups containing from x to y carbons in the chain, and the straight-chain alkyl groups and branched-chain alkyl groups include haloalkyl groups such as trifluoromethyl and 2,2,2-trifluoroethyl, etc.). When the group is in a terminal position, C 0 alkyl represents hydrogen, and when the group is internal, C 0 alkyl represents a bond. The terms "C 2-y alkenyl" and "C 2-y alkynyl" refer to substituted or unsubstituted unsaturated aliphatic hydrocarbon groups that are similar to the alkyls described above in length and possible substituents, but contain at least one double bond or triple bond, respectively.

[0044] As used herein, the term "alkylamino" refers to an amino group substituted with at least one alkyl group.

[0045] As used herein, the term "alkylthio" refers to a thiol group substituted with an alkyl group and can be represented by the general formula alkylS-.

[0046] As used herein, the term "alkynyl" refers to an aliphatic hydrocarbon group containing at least one triple bond and is intended to include both "unsubstituted alkynyl" and "substituted alkynyl", where the latter refers to an alkynyl moiety having substituents replacing the hydrogens on one or more carbons of the alkynyl group. Such substituents can be present on one or more carbons that are or are not included in one or more triple bonds. In addition, such substituents include all those contemplated for alkyl groups as described above, except where stability is prohibited. For example, it is contemplated that the alkynyl group is substituted with one or more alkyl, carbocyclic, aryl, heterocyclic, or heteroaryl groups.

[0047] As used herein, the term "amide" refers to the group

[0048]

[0049] where each R 10 independently represents hydrogen or a hydrocarbon group, or two Rs 10 together with the N atom to which the two Rs 10 are attached form a heterocycle having from 4 to 8 atoms in the ring structure.

[0050] The terms "amine" and "amino" are well recognized in the art and refer to both unsubstituted and substituted amines and their salts, for example, a moiety represented by the following formula:

[0051]

[0052] where each R 10 independently represents hydrogen or a hydrocarbon group, or two Rs 10 together with the N atom to which the two Rs 10 are attached form a heterocycle having from 4 to 8 atoms in the ring structure. As used herein, the term "aminoalkyl" refers to an alkyl group substituted with an amino group.

[0053] As used herein, the term "aralkyl" refers to an alkyl group substituted with an aryl group.

[0054] As used herein, the term "aryl" includes substituted or unsubstituted monocyclic aromatic groups, wherein each atom of the ring is carbon. Preferably, the ring is a 5- to 7-membered ring, more preferably a 6-membered ring. The term "aryl" also includes polycyclic ring systems having two or more cyclic rings, wherein two or more carbons are common to two adjacent rings, wherein at least one of the rings is aromatic; for example, the other cyclic rings can be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclic. Aryl groups include benzene, naphthalene, phenanthrene, phenol, aniline, and the like.

[0055] The term "carbamate" is well recognized in the art and refers to the following groups:

[0056]

[0057] wherein R 9 and R 10 independently represent hydrogen or a hydrocarbyl group (such as an alkyl group), or R 9 and R 10 together with one or more intervening atoms form a heterocycle having from 4 to 8 atoms in the ring structure.

[0058] As used herein, the terms "carbocyclic" and "carbocyclic-" refer to saturated or unsaturated rings, wherein each atom of the ring is carbon. The term carbocyclic includes both aromatic carbocyclic and non-aromatic carbocyclic rings. Non-aromatic carbocyclic rings include both cycloalkane rings (wherein all carbon atoms are saturated) and cycloalkene rings (which contain at least one double bond).

[0059] The term "carbocyclic" includes 5-7 membered monocyclics and 8-12 membered bicyclics. Each ring of the bicyclic carbocyclic can be selected from saturated rings, unsaturated rings, and aromatic rings. Carbocyclic includes bicyclic molecules wherein one, two, three, or more atoms are shared between the two rings. The term "fused carbocyclic" refers to a bicyclic carbocyclic wherein each of the rings shares two adjacent atoms with the other ring. Each ring in the fused carbocyclic can be selected from saturated rings, unsaturated rings, and aromatic rings. In an exemplary embodiment, an aromatic ring (e.g., phenyl) can be fused to a saturated or unsaturated ring (e.g., cyclohexane, cyclopentane, or cyclohexene). Any combination of saturated bicyclics, unsaturated bicyclics, and aromatic bicyclics is included in the definition of carbocyclic where valence allows. Exemplary "carbocyclics" include cyclopentane, cyclohexane, bicyclo[2.2.1]heptane, 1,5-cyclooctadiene, 1,2,3,4-tetrahydronaphthalene, bicyclo[4.2.0]oct-3-ene, naphthalene, and adamantane. Exemplary fused carbocyclics include decahydronaphthalene, naphthalene, 1,2,3,4-tetrahydronaphthalene, bicyclo[4.2.0]octane, 4,5,6,7-tetrahydro-1H-indene, and bicyclo[4.1.0]hept-3-ene. A "carbocyclic" can be substituted at any one or more positions capable of bearing a hydrogen atom.

[0060] The "cycloalkyl" group is a fully saturated cycloalkane. "Cycloalkyl" includes monocyclic and bicyclic rings. Typically, unless otherwise defined, a monocyclic cycloalkyl group has from 3 to about 10 carbon atoms, more typically 3 to 8 carbon atoms. The second ring of a bicyclic cycloalkyl can be selected from a saturated ring, an unsaturated ring, and an aromatic ring. Cycloalkyl includes bicyclic molecules in which one, two, three, or more atoms are shared between the two rings. The term "fused cycloalkyl" refers to a bicyclic cycloalkyl in which each of the rings shares two adjacent atoms with the other ring. The second ring of a fused bicyclic cycloalkyl can be selected from a saturated ring, an unsaturated ring, and an aromatic ring. The "cycloalkenyl" group is a cycloalkane containing one or more double bonds.

[0061] As used herein, the term "carbocyclic alkyl" refers to an alkyl group substituted with a carbocyclic group.

[0062] The term "carbonate" is well recognized in the art and refers to the group -OCO 2 -R 10 where R 10 represents a hydrocarbyl group.

[0063] As used herein, the term "carboxy" refers to the group represented by the formula -CO 2 H.

[0064] As used herein, the term "ester" refers to the group -C(O)OR 10 where R 10 represents a hydrocarbyl group.

[0065] As used herein, the term "ether" refers to a hydrocarbyl group linked to another hydrocarbyl group through oxygen. Accordingly, an ether substituent of a hydrocarbyl group can be hydrocarbyl-O-. An ether can be symmetric or asymmetric. Examples of ethers include, but are not limited to, heterocyclic-O-heterocyclic and aryl-O-heterocyclic. Ethers include the "alkoxyalkyl" group, which can be represented by the general formula alkyl-O-alkyl.

[0066] As used herein, the terms "halo" and "halogen" mean the halogen group and include chloro, fluoro, bromo, and iodo.

[0067] As used herein, the terms "hetaralkyl" and "heteroaralkyl" refer to an alkyl group substituted with a heteroaryl group.

[0068] As used herein, the term "heteroalkyl" refers to a saturated or unsaturated chain of carbon atoms and at least one heteroatom, where no two heteroatoms are adjacent.

[0069] The terms "heteroaryl" and "hetaryl" include substituted or unsubstituted aromatic monocyclic structures (preferably 5- to 7-membered rings, more preferably 5- to 6-membered rings), the ring structure of which contains at least one heteroatom (preferably one to four heteroatoms, more preferably one or two heteroatoms). The terms "heteroaryl" and "hetaryl" also include polycyclic ring systems having two or more cyclic rings, wherein two or more carbons are common to two adjacent rings, and wherein at least one of the rings is heteroaromatic. For example, the other cyclic rings can be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclic. Heteroaryl groups include, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrazine, pyridazine, and pyrimidine, etc.

[0070] As used herein, the term "heteroatom" means an atom of any element other than carbon or hydrogen. Preferred heteroatoms are nitrogen, oxygen, and sulfur.

[0071] The terms "heterocyclic group", "heterocycle", and "heterocyclic" refer to substituted or unsubstituted non-aromatic ring structures (preferably 3- to 10-membered rings, more preferably 3- to 7-membered rings), the ring structure of which contains at least one heteroatom, preferably one to four heteroatoms, more preferably one or two heteroatoms. The terms "heterocyclic group" and "heterocyclic" also include polycyclic ring systems having two or more cyclic rings, wherein two or more carbons are common to two adjacent rings, and wherein at least one of the rings is heterocyclic. For example, the other cyclic rings can be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclic. Heterocyclic groups include, for example, piperidine, piperazine, pyrrolidine, morpholine, lactone, lactam, etc.

[0072] As used herein, the term "heterocyclic alkyl" refers to an alkyl group substituted with a heterocyclic group.

[0073] As used herein, the term "hydrocarbyl" refers to a group that is bonded through a carbon atom that does not have an =O or =S substituent, and that generally has at least one carbon-hydrogen bond and a predominantly carbon backbone, but that may optionally contain heteroatoms. Thus, groups such as methyl, ethoxyethyl, 2-pyridyl, and trifluoromethyl are considered to be hydrocarbyl groups for the purposes of this application, but substituents such as acetyl (which has an =O substituent on the connecting carbon) and ethoxy (which is connected through oxygen, rather than carbon) are not hydrocarbyl groups for the purposes of this application. Hydrocarbyl groups include, but are not limited to, aryl, heteroaryl, carbocyclic, heterocyclic, alkyl, alkenyl, alkynyl, and combinations thereof.

[0074] As used herein, the term "hydroxyalkyl" refers to an alkyl group substituted with a hydroxy group.

[0075] When used in combination with a chemical moiety (e.g., acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy), the term "lower" means a group containing ten or fewer non-hydrogen atoms (preferably six or fewer) in the substituent. For example, "lower alkyl" refers to an alkyl group containing ten or fewer carbon atoms (preferably six or fewer). In certain embodiments, the acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy substituents as defined herein are lower acyl, lower acyloxy, lower alkyl, lower alkenyl, lower alkynyl, or lower alkoxy, whether they appear alone or in combination with other substituents, as in the description of hydroxyalkyl and aralkyl (in the case of aralkyl, for example, when counting the carbon atoms in the alkyl substituent, the atoms within the aryl group are not counted).

[0076] The terms "polycyclic group", "polycycle", and "polycyclic" refer to two or more rings (e.g., cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclic), where two or more atoms are common to two adjacent rings, e.g., the rings are "fused rings". Each of the rings in the polycycle can be substituted or unsubstituted. In certain embodiments, each ring in the polycycle contains from 3 to 10 atoms, preferably from 5 to 7 atoms, in the ring.

[0077] The term "silyl" refers to a silicon moiety having three hydrocarbon moieties attached thereto.

[0078] The term "substituted" refers to a moiety having a substituent that replaces a hydrogen on one or more carbons of the backbone. It will be understood that "substitution" or "substituted with" encompasses the implicit proviso that such substitution is in accordance with the allowed valences of the atoms being substituted and the substituent and that the substitution results in a stable compound (e.g., the compound does not spontaneously undergo transformations such as by rearrangement, cyclization, elimination, etc.). As used herein, the term "substituted" is intended to include all allowed substituents of an organic compound. In a broader aspect, allowed substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of an organic compound. For suitable organic compounds, the allowed substituents can be one or more and can be the same or different. For the purposes of the present invention, a heteroatom, such as nitrogen, can have a hydrogen substituent and / or any allowed substituent of the organic compounds described herein that satisfies the valence of the heteroatom. Substituents can include any of the substituents described herein, e.g., a halogen, hydroxyl, carbonyl (such as carboxyl, alkoxycarbonyl, formyl, or acyl), thiocarbonyl (such as thioester, thioacetate, or thiocarboxylate), alkoxy, phosphoryl, phosphate, phosphonate, phosphinate, amino, amido, amidine, imine, cyano, nitro, azide, mercapto, alkylthio, sulfate, sulfonate, sulfamoyl, sulfinylamino, sulfonyl, heterocyclic group, aralkyl, or an aromatic moiety or heteroaromatic moiety. Those skilled in the art will understand that a substituent can itself be substituted (if appropriate). Unless expressly stated as "unsubstituted", a reference herein to a chemical moiety is understood to include substituted variants. For example, a reference to an "aryl" group or moiety implicitly includes both substituted and unsubstituted variants.

[0079] The term "sulfate" is well recognized in the art and refers to the group -OSO 3 H or a pharmaceutically acceptable salt of the group -OSO 3 H.

[0080] The term "sulfonamide" is well recognized in the art and refers to a group represented by the general formula

[0081]

[0082] wherein R 9 and R 10 independently represent hydrogen or a hydrocarbon group (such as an alkyl group), or R 9 and R 10 together with one or more intervening atoms form a heterocycle having from 4 to 8 atoms in the ring structure.

[0083] The term "sulfoxide" is well recognized in the art and refers to the group -S(O)-R 10 wherein R10 represents a hydrocarbon group.

[0084] The term "sulfonate" is well - recognized in the art and refers to the group SO 3 H or a pharmaceutically acceptable salt of the group SO 3 H.

[0085] The term "sulfone" is well - recognized in the art and refers to the group - S(O) 2 -R 10 wherein R 10 represents a hydrocarbon group.

[0086] As used herein, the term "thioalkyl" refers to an alkyl group substituted with a thiol group.

[0087] As used herein, the term "thioester" refers to the group - C(O)SR 10 or - SC(O)R 10 wherein R 10 represents a hydrocarbon group.

[0088] As used herein, the term "thioether" is equivalent to an ether in which sulfur replaces oxygen.

[0089] The term "urea" is well - recognized in the art and can be represented by the following general formula

[0090]

[0091] wherein R 9 and R 10 independently represent hydrogen or a hydrocarbon group (such as an alkyl group), or any occurrence of R 9 together with R 10 and one or more intervening atoms forms a heterocyclic ring having from 4 to 8 atoms in the ring structure.

[0092] The term "protecting group" refers to a group of atoms that, when attached to a reactive functional group in a molecule, masks, reduces, or hinders the reactivity of the functional group. Typically, during synthesis, protecting groups can be selectively removed as needed. Examples of protecting groups can be found in Greene and Wuts, Protective Groups in Organic Chemistry, 3 rdfound in Ed., 1999, John Wiley & Sons, NY and Harrison et al., Compendium of Synthetic Organic Methods, Vols. 1 - 8, 1971 - 1996, John Wiley & Sons, NY. Representative nitrogen protecting groups include, but are not limited to, formyl, acetyl, trifluoroacetyl, benzyl, benzyloxycarbonyl (“CBZ”), tert - butyloxycarbonyl (“Boc”), trimethylsilyl (“TMS”), 2 - trimethylsilylethanesulfonyl (“TES”), triphenylmethyl and substituted triphenylmethyl groups, allyloxycarbonyl, 9 - fluorenylmethoxycarbonyl (“FMOC”), nitroveratryloxycarbonyl (“NVOC”), etc. Representative hydrocarbyl protecting groups include, but are not limited to, those in which the hydrocarbyl group is acylated (esterified) or alkylated, such as benzyl and triphenylmethyl ethers, as well as alkyl ethers, tetrahydropyranyl ethers, trialkylsilyl ethers (e.g., TMS or TIPS groups), ethylene glycol ethers, such as ethylene glycol derivatives, propylene glycol derivatives, and allyl ethers.

[0093] In certain embodiments, the compounds of the invention can be racemic. In certain embodiments, the compounds of the invention can be enriched in one enantiomer. For example, the compounds of the invention can have an ee of greater than about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, or even about 95% or greater. In certain embodiments, the compounds of the invention can have more than one stereocenter. In certain such embodiments, the compounds of the invention can be enriched in one or more diastereomers. For example, the compounds of the invention can have a de of greater than about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, or even about 95% or greater.

[0094] In certain embodiments, a therapeutic formulation can be enriched to predominantly provide one enantiomer of a compound (e.g., a compound of formula (1)). An enantiomerically enriched mixture can include, for example, at least about 60 mol percent of one enantiomer, or more preferably at least about 75, about 90, about 95, or even about 99 mol percent. In certain embodiments, a compound enriched in one enantiomer is substantially free of the other enantiomer, where substantially free means, for example, in a composition or mixture of compounds, the substance in question is present in less than about 10%, or less than about 5%, or less than about 4%, or less than about 3%, or less than about 2%, or less than about 1% compared to the amount of the other enantiomer. For example, if a composition or mixture of compounds contains about 98 grams of a first enantiomer and about 2 grams of a second enantiomer, it is said to contain about 98 mol percent of the first enantiomer and only about 2% of the second enantiomer.

[0095] In certain embodiments, a therapeutic formulation can be enriched to predominantly provide one diastereomer of a compound (e.g., a compound of formula (1)). A diastereomerically enriched mixture can include, for example, at least about 60 mol percent of one diastereomer, or more preferably at least about 75, about 90, about 95, or even about 99 mol percent of one diastereomer.

[0096] The term "subject" (to which administration is contemplated) includes (but is not limited to) humans (i.e., males or females of any age group, e.g., pediatric subjects (e.g., infants, children, adolescents) or adult subjects (e.g., young adults, middle-aged adults or elderly adults)) and / or other primates (e.g., cynomolgus monkeys, rhesus monkeys); mammals (including commercially relevant mammals such as cows, pigs, horses, sheep, goats, cats, and / or dogs); and / or birds (including commercially relevant birds such as chickens, ducks, geese, quails, and / or turkeys). Preferred subjects are humans.

[0097] As used herein, a therapeutic agent that "prevents" a disorder or condition refers to a compound that, in a statistical sample, reduces the occurrence of the disorder or condition in a treated sample relative to an untreated control sample, or delays the onset of one or more symptoms of the disorder or condition or reduces the severity of one or more symptoms of the disorder or condition relative to an untreated control sample.

[0098] The term "treatment" encompasses prophylactic treatment of a disease and / or therapeutic treatment. The term "prophylactic or therapeutic" treatment is well recognized in the art and encompasses administering to a subject one or more of the disclosed compositions. If administered prior to the clinical manifestation of a deleterious condition (e.g., a disease or other deleterious state of the subject), the treatment is prophylactic (i.e., it protects the subject from developing the deleterious condition), while if it is administered after the manifestation of the deleterious condition, the treatment is therapeutic (i.e., it is intended to reduce, alleviate or stabilize an existing deleterious condition or its side effects).

[0099] The term "prodrug" is intended to encompass compounds (e.g., compounds of formula (I)) that are converted into a therapeutic active agent of the present invention under physiological conditions. Common methods for making prodrugs involve including one or more selected moieties that are hydrolyzed under physiological conditions to expose the desired molecule. In other embodiments, the prodrug is converted by the enzymatic activity of the subject. For example, esters or carbonates (e.g., esters or carbonates of alcohols or carboxylic acids) are preferred prodrugs of the present invention. In certain embodiments, some or all of the compounds of formula I in the above dosage forms can be replaced with the corresponding suitable prodrugs, e.g., where a hydroxyl group in the parent compound is present as an ester or carbonate or carboxylic acid.

[0100] As used herein, the term "effective amount" refers to an amount sufficient to achieve the desired biological effect. As used herein, "therapeutically effective amount" refers to an amount sufficient to achieve the desired therapeutic effect. By way of example, a therapeutically effective amount may refer to an amount sufficient to improve at least one sign or symptom of cancer.

[0101] "Response" to a treatment method may particularly include reduction or improvement of negative symptoms, slowing of the progression of a disease or its symptoms, increase in beneficial symptoms or clinical outcomes, reduction of side effects, disease stabilization, partial or complete cure of the disease.

[0102] Overview

[0103] In certain embodiments, there is provided herein a compound of formula (I):

[0104]

[0105] wherein

[0106] each Q, Y and Z is independently selected from N and C, and X is N or C-R a ;

[0107] provided that at least one of Q, X, Y and Z is N, and

[0108] each dashed bond is independently a single bond or a double bond such that the bicyclic ring formed thereby is a heteroaryl;

[0109] R 1 is selected from alkyl, alkenyl, alkynyl, amino, acylamino, cycloalkyl, cycloalkylalkyl, heterocyclic group, heterocyclic group alkyl, aryl, aralkyl, heteroaryl and heteroarylalkyl;

[0110] or

[0111] a) W is N, R 2 does not exist, and A 1 and A 2 are each CH 2 ;

[0112] b) W is C, R 2 is selected from H, halogen, CN, alkyl, alkoxy, hydroxyl, acyloxy and amino, and A 1 and A 2 are each independently selected from CH 2 and O; or

[0113] c) W is C, and R 1 and R 2 together form =CH 2 -acylamino, cycloalkyl and heterocyclic group, and A 1 and A 2 are each independently selected from CH 2 and O;

[0114] R a is selected from H, halogen, CN and alkyl;

[0115] R 9 is selected from H, halogen, alkyl, alkoxy, hydroxyl, acyloxy and amino;

[0116] R 10 is H or alkoxy;

[0117] R 11 is aryloxy, heteroaryloxy, aralkyl, alkoxycarbonyl, ureido or -C(O)-aryl; and

[0118] R 12 is selected from H, halogen, CN, alkyl, alkoxy, hydroxyl and acyloxy;

[0119] provided that the compound of formula (I) is not

[0120] and if W is C, R 2 、R 9 and R 12 are each hydrogen, and R 11 is unsubstituted phenoxy, then

[0121] R 1 is not -NHCH2 OH, -NHCH 2 COOH, -NHCH 2 CONH 2 , -NHCH 2 CH 2 NH 2 , -NH(CH 2 ) 2 N(Me) 2 , -NHCH 2 -pyridyl, -NHCO-pyridyl, -C(O)OEt, -NH-tetrahydropyran, -NHCH 2 pyrrolidinyl, -NH(CH 2 ) 3 imidazolyl, -NH-pyrrolidinoneyl,

[0122] wherein R 13 is selected from H, ethyl, isopropyl, tert-butyl, -CH 2 OCH 3 , -CH 2 CH 2 OH and -CH 2 CH 2 OCH 3 ; and if W is N, R 9 and R 12 are each hydrogen, and R 11 is unsubstituted phenoxy, then R 1 is not piperidinyl.

[0123] In certain embodiments, the compound has the following structure: wherein R 2 is H.

[0124] In other embodiments, the compound has the following structure: wherein R 2 is H.

[0125] In other embodiments, the compound has the following structure: wherein R 2 is H.

[0126] In other embodiments, the compound has the following structure: wherein R 2 is H.

[0127] In some embodiments, at least two of Q, X, Y, and Z are N. In some such embodiments, at least three of Q, X, Y, or Z are N. In some such embodiments, Q, X, Y, and Z are all N.

[0128] In some embodiments, at least Q is N.

[0129] In some embodiments, at least X is N. In some such embodiments, at least X and Z are N.

[0130] In some embodiments, at least Y is N. In some such embodiments, at least X and Y are N.

[0131] In some embodiments, at least Z is N.

[0132] In some embodiments, R 1 is V is N or CH; T is N or CH; R 3 is an alkyl; and R 4 is H or an alkyl.

[0133] In some embodiments, R 1 is V is N or CH; T is N or CH; R 3 is an alkyl; and R 4 is H or an alkyl.

[0134] In some embodiments, Q is C, X is C-R a , R a is H, Y is N and Z is C; Q is C, X is C-R a , R a is a halogen, Y is N and Z is C; Q is C, X is N, Y is C and Z is N; Q is C, X is N, Y is N and Z is C; Q is N, X is C-R a , R a is H, Y is C and Z is C; Q is C, X is C-R a , R a is H, Y is C and Z is N; or Q is C, X is C-R a , R a is H, Y is N and Z is C.

[0135] In some embodiments, Q is C, X is N, Y is C and Z is N; Q is C, X is C-R a , R a is H, Y is N and Z is C; or Q is C, X is N, Y is N and Z is C.

[0136] In some embodiments, A 1 and A 2 are both CH 2 . In other embodiments, A 1 and A 2 are both O.

[0137] In some embodiments, W is N and R 2 is absent. In other embodiments, W is C.

[0138] In some embodiments, R 2 is selected from H, hydroxy, and amino.

[0139] In some embodiments, R 1 is selected from alkyl, amino, cycloalkyl, heterocyclic, heterocyclic alkyl, and heteroaryl. In certain preferred embodiments, R 1 is a 6-membered heterocyclic group, preferably containing 2 nitrogen ring atoms. In particularly preferred embodiments, R 1 is a 1,4-piperazinyl ring.

[0140] In certain such embodiments, R 1 is selected from wherein V is N or CH; T is N and R 4 ' is present or T is C; J is selected from sulfonyl, C(H)sulfinylamino, and >P(=O)-alkyl; R 3 is selected from H, halogen, CN, alkyl, alkoxy, hydroxy, acyloxy, amino, acylamino, cycloalkyl, heterocyclic, carboxyl, and alkoxycarbonyl; R 3 ' is H or alkyl, or R 3 and R 3 ' together with the carbon atom to which it is attached form a cycloalkyl; R 4 is selected from H, halogen, CN, alkyl, alkoxy, acyloxy, amino, and acylamino; R 4 ' is H or alkyl; R 5 is H or alkyl; R 6 and R 6 ' are each H; R 6 and R 6 ' together with the carbon atom to which it is attached form a carbonyl (oxo); R 7 is H or alkyl; and R 8 is H, hydroxy, acyloxy, or amino.

[0141] In some such embodiments, R 1 is In some such embodiments, V is N. In other such embodiments, V is CH.

[0142] In some such embodiments, T is N. In other such embodiments, T is C.

[0143] In certain embodiments, W is C, R 2 is H, V is CH, and T is N. In other embodiments, W is C, R 2is H, V is N and T is C. In other embodiments, W is C, R 2 is H, V is N, and T is N. In still other embodiments, W is N, R 2 is absent, V is CH, and T is N.

[0144] In some embodiments, R 3 is selected from H, alkyl, amino, cycloalkyl, heterocyclic, carboxyl, and alkoxycarbonyl. In certain such embodiments, R 3 is H. In other such embodiments, R 3 is alkyl. In some such embodiments, R 3 is methyl.

[0145] In some embodiments, each R 4 is independently selected from H, alkyl, amino, and amido. In certain such embodiments, R 4 is H. In other such embodiments, R 4 is alkyl. In some such embodiments, R 4 is methyl.

[0146] In some embodiments, R 5 is H.

[0147] In certain embodiments, R 9 is selected from halogen, alkyl, alkoxy, hydroxy, acyloxy, and amino, preferably alkyl or halogen, most preferably halogen. In certain most preferred embodiments, R 9 is F.

[0148] In some embodiments, R 12 is H or halogen.

[0149] In some embodiments, R 11 is selected from amido, wherein B is selected from -O-, CH 2 , CHOH, NH, N(C 1-6 alkyl), and carbonyl; K 1 and K 2 are independently selected from N and CR 13 ; D is selected from -CH 2 -C(O)-, -NH-C(O)-, N(C 1-6 alkyl)-C(O), and -CH 2 S(O) 2 -; R 13 is selected from H, halogen, CN, alkyl, alkoxy, hydroxy, acyloxy, alkoxycarbonyl, amino, amido, and alkoxycarbonyl; R 14Selected from H, halogen, CN, alkyl, alkoxy, hydroxy, acyloxy, alkoxycarbonyl, amino, acylamino and alkoxycarbonyl; R 15 Selected from H, halogen, CN, alkyl, alkoxy, hydroxy, acyloxy, alkoxycarbonyl, amino, acylamino and alkoxycarbonyl; and R 16 Selected from H, halogen, CN, alkyl, alkoxy, hydroxy, acyloxy, amino, acylamino and alkoxycarbonyl.

[0150] In some embodiments, B is -O-.

[0151] In some embodiments, K 1 is CR 13 . In some such embodiments, R 13 is selected from H, halogen, CN and alkyl. In certain such embodiments, R 13 is H or F.

[0152] In some embodiments, R 14 is H or alkyl.

[0153] In some embodiments, R 15 is H or halogen.

[0154] In some embodiments, D is -NHC(O)-.

[0155] In some embodiments, R 16 is H or alkyl.

[0156] In some embodiments, R 11 is wherein B is O; K 1 and K 2 are each CH; R 14 is H; and R 15 is H. In other embodiments, R 11 is wherein K 1 and K 2 are each N; and R 14 and R 15 are each H. In other embodiments, R 11 is wherein D is NH; and R 16 is alkyl.

[0157] In certain preferred embodiments, the compounds of formula (I) are selected from

[0158]

[0159] In some such preferred embodiments, the compounds of formula (I) are selected from

[0160]

[0161] For example, the compound of formula (I) can be Alternatively, the compound of formula (I) can be Alternatively, the compound of formula (I) can be

[0162] Similarly, the compound of formula (I) can be

[0163] In certain embodiments, provided herein is a method for co-inhibiting HCK and BCL-2 in cells, which comprises contacting the cells with a compound of formula (I) and a BCL-2 inhibitor. In certain preferred embodiments, the compound is an HCK inhibitor. In some such embodiments, the HCK inhibitor is a dual HCK / FLT3-ITD inhibitor. In certain embodiments, the method further comprises contacting the cells with an FLT3-ITD inhibitor.

[0164] In certain embodiments, provided herein is a method for killing cells having an FLT3-ITD mutation, which comprises contacting the cells with a compound of formula (I) and a BCL-2 inhibitor. In certain preferred embodiments, the compound is an HCK inhibitor. In some such embodiments, the HCK inhibitor is a dual HCK / FLT3-ITD inhibitor. In certain embodiments, the method further comprises contacting the cells with an FLT3-ITD inhibitor.

[0165] In certain embodiments, provided herein is a method for treating acute myeloid leukemia, which comprises co-administering to a subject a compound of formula (I) and a BCL-2 inhibitor. In some such embodiments, the subject has FLT3-ITD+ acute myeloid leukemia. In certain such embodiments, the subject has malignant hematopoiesis and / or non-malignant multi-lineage hematopoiesis, characterized by having one or more mutations in a gene selected from the following: DNMT3A, IDH2, IDH1, NPM1, TET2, CEBPA, ASXL1, EZH2, SETBP1, SMC3, KIT, NRAS, and WT1. In some such embodiments, the compound of formula (I) is an HCK inhibitor. In certain such embodiments, the method further comprises co-administering an FLT3-ITD inhibitor. In other such embodiments, the HCK inhibitor is a dual HCK / FLT3-ITD inhibitor.

[0166] In certain embodiments, the HCK inhibitor, the FLT3-ITD inhibitor, and the BCL-2 inhibitor are administered simultaneously or sequentially in separate unit dosage forms. In other embodiments, the method comprises administering a single unit dosage form that comprises an HCK inhibitor, a BCL-2 inhibitor, and a pharmaceutically acceptable carrier, adjuvant, or vehicle. In some such embodiments, the single unit dosage form further comprises an FLT3-ITD inhibitor, or wherein the HCK inhibitor is a dual HCK / FLT3-ITD inhibitor.

[0167] In certain embodiments, the FLT3-ITD inhibitor is selected from AC220, sorafenib, PKC412, CEP-701, UNC2025, MLN518, KW-2449, and AMG-925, sunitinib, SU5614, AC2206, crenolanib, and PLX3397. In some such embodiments, the BCL-2 inhibitor is selected from AT-101, TW-37, TM-1206, gossypolic acid, gossypolonic acid, apogossypol, apogossypolone, A385358, ABT-737, ABT-263, ABT-199, WEHI-539, BXI-61, BXI-72, obatoclax, JY-1-106, and SAHB peptide. In some such embodiments, the BCL-2 inhibitor is selected from gossypol, obatoclax, ABT-737, ABT-199, and ABT-263. In some such embodiments, the BCL-2 inhibitor is ABT-199. In some such embodiments, the HCK inhibitor is (1) and the BCL-2 inhibitor is ABT-199. In other such embodiments, the HCK inhibitor is (2) and the BCL-2 inhibitor is ABT-199. In other such embodiments, the HCK inhibitor is (3) and the BCL-2 inhibitor is ABT-199. In other such embodiments, the HCK inhibitor is (4) and the BCL-2 inhibitor is ABT-199. In other such embodiments, the FLT3-ITD inhibitor is AC220 and the BCL-2 inhibitor is ABT-199. In other embodiments, the FLT3-ITD inhibitor is SU5614 and the BCL-2 inhibitor is ABT-737. In some such embodiments, the HCK inhibitor and / or the FLT3-ITD inhibitor and / or the BCL-2 inhibitor are each present in a pharmaceutically acceptable salt form. In some such embodiments, the HCK inhibitor and / or the FLT3-ITD inhibitor and / or the BCL-2 inhibitor are each present in a pharmaceutically acceptable composition form.

[0168] Compositions and salts

[0169] In some embodiments, the disclosed compounds may be in the form of a pharmaceutically acceptable composition. Disclosed herein are pharmaceutical compositions comprising an FLT3-ITD inhibitor and / or an HCK inhibitor and / or a BCL-2 inhibitor as described herein and a pharmaceutically acceptable carrier.

[0170] The term "pharmaceutically acceptable carrier, adjuvant or vehicle" refers to a non-toxic carrier, adjuvant or vehicle that does not destroy the pharmacological activity of the compounds formulated therewith. Pharmaceutically acceptable carriers, adjuvants or vehicles useful in the compositions of the present invention include, but are not limited to, ion exchangers; alumina; aluminum stearate; lecithin, serum proteins such as human serum albumin; buffering substances such as phosphates; glycine; sorbic acid; potassium sorbate; mixtures of partial glycerides of saturated vegetable fatty acids; water; salts or electrolytes such as protamine sulfate; disodium hydrogen phosphate; potassium hydrogen phosphate; sodium chloride; zinc salts; colloidal silica; magnesium trisilicate; polyvinylpyrrolidone; cellulose-based substances; polyethylene glycol; sodium carboxymethylcellulose; polyacrylates; waxes; polyethylene-polypropylene oxide-block polymers; polyethylene glycol and lanolin. Other pharmaceutically acceptable carriers, adjuvants or vehicles include water, physiological saline and dimethyl sulfoxide as well as other hydrophobic or hydrophilic solvents.

[0171] Examples of suitable aqueous and non-aqueous carriers that may be used in pharmaceutical compositions include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol and the like) and suitable mixtures thereof, vegetable oils (such as olive oil) and injectable organic esters (such as ethyl oleate). Appropriate fluidity may be maintained, for example, by the use of coating materials such as lecithin, by maintaining the desired particle size in the case of dispersions and by the use of surfactants.

[0172] Such compositions may also contain adjuvants such as preservatives, wetting agents, emulsifying agents, dispersing agents, lubricants and / or antioxidants. Prevention of the action of microorganisms on the compounds described herein may be ensured by including various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid and the like. It may also be necessary to include isotonic agents such as sugars, sodium chloride and the like in the composition.

[0173] Methods of preparing such formulations or compositions include the step of combining the compounds described herein with a carrier and optionally one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing into association the compounds as disclosed herein with a liquid carrier or a finely divided solid carrier or both and, if necessary, then shaping the product.

[0174] Formulations for such pharmaceutical compositions are well known in the art. See, for example, Anderson, Philip O.; Knoben, James E.; Troutman, William G., eds., Handbook of Clinical Drug Data, 10th Edition, McGraw-Hill, 2002; Pratt and Taylor, eds., Principles of Drug Action, 3rd Edition, Churchill Livingston, New York, 1990; Katzung, ed., Basic and Clinical Pharmacology, 9th Edition, McGraw Hill, 2003; Goodman and Gilman, eds., The Pharmacological Basis of Therapeutics, 10th Edition, McGraw Hill, 2001; Remington’s Pharmaceutical Sciences, 20th Edition, Lippincott Williams & Wilkins., 2000; Martindale, The Extra Pharmacopoeia, 32nd Edition (The Pharmaceutical Press, London, 1999); each of which is incorporated herein by reference in its entirety. The use of excipients is contemplated within the scope of the present invention, unless any well-known excipient medium is incompatible with the compounds provided herein, such as producing any undesirable biological effects or otherwise interacting adversely with any other component of the pharmaceutically acceptable composition.

[0175] The present disclosure provides pharmaceutically acceptable salts, which refer to those salts that are suitable, within the scope of reasonable medical judgment, for contact with the tissues of a subject without excessive toxicity, irritation, allergic response, and similar reactions and that satisfy a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, Berge et al. described pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences (1977) 66:1-19. Pharmaceutically acceptable salts of the compounds provided herein include those derived from suitable inorganic acids, organic acids, and bases. Examples of pharmaceutically acceptable non-toxic acid addition salts are salts formed by reacting an amino group with an inorganic acid such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid, or with an organic acid such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid, or salts formed by using other methods used in the art such as ion exchange. Other pharmaceutically acceptable salts include adipates, alginates, ascorbates, aspartates, benzenesulfonates (benzenesulfonate / besylate), benzoates, bisulfates, borates, butyrates, camphorates, camphorsulfonates, citrates, cyclopentanepropionates, digluconates, dodecyl sulfates, ethanesulfonates, formates, fumarates, glucoheptanoates, glycerophosphates, glucuronates, hemisulfates, heptanoates, hexanoates, hydroiodides, 2-hydroxyethanesulfonates, lactates, lactobionates, laurates, lauryl sulfates, malates, maleates, malonates, methanesulfonates, 2-naphthalenesulfonates, nicotinates, nitrates, oleates, oxalates, palmitates, pamoates, pectates, persulfates, 3-phenylpropionates, phosphates, picrates, pivalates, propionates, stearates, succinates, sulfates, tartrates, thiocyanates, tosylates, undecanoates, valerates, and the like. In some embodiments, the organic acids from which the salts can be derived include, for example, acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, lactic acid, trifluoroacetic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, and the like.

[0176] The salts can be prepared in situ during the isolation and purification of the disclosed compounds, or separately, for example, by reacting the free base or free acid of the compound with a suitable base or acid, respectively. Pharmaceutically acceptable salts derived from suitable bases include alkali metals, alkaline earth metals, ammonium, and N + (C 1-4 alkyl) 4Salts. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, iron, zinc, copper, manganese, aluminum, and similar metals. Where appropriate, other pharmaceutically acceptable salts include non-toxic ammonium, quaternary ammonium, and amine cations formed with counterions such as halide ions, hydroxide, carboxylate, sulfate, phosphate, nitrate, lower alkyl sulfonate, and aryl sulfonate. Organic bases from which salts can be derived include, for example, primary, secondary, and tertiary amines, substituted amines (including naturally occurring substituted amines), cyclic amines, basic ion exchange resins, and their analogs, such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, and ethanolamine. In some embodiments, the pharmaceutically acceptable base addition salts are selected from ammonium salts, potassium salts, sodium salts, calcium salts, and magnesium salts.

[0177] Pharmaceutically acceptable acid addition salts can also exist in the form of various solvates with solvents such as water, methanol, ethanol, dimethylformamide, and their analogs. Mixtures of such solvates can also be prepared. The source of such solvates can come from the solvents of crystallization, which are inherent in the solvents used for preparation or crystallization or added to such solvents.

[0178] The pharmaceutical composition (formulation) can be administered to a subject by any of a variety of routes of administration, including, for example: orally (e.g., as a large dose liquid medicine, lozenge, capsule (including dispersed capsule and gelatin capsule), bolus, powder, granule, paste coated on the tongue) in the form of an aqueous solution or non-aqueous solution or suspension; absorption through the oral mucosa (e.g., sublingually); subcutaneously; transdermally (e.g., in the form of a patch applied to the skin); and topically (e.g., in the form of a cream, ointment, or spray applied to the skin). The compound can also be formulated for inhalation. In certain embodiments, the compound can simply be dissolved or suspended in sterile water. Details of the appropriate route of administration and the compositions suitable for it can be found, for example, in U.S. Patent Nos. 6,110,973, 5,763,493, 5,731,000, 5,541,231, 5,427,798, 5,358,970, and 4,172,896, and the patents cited therein.

[0179] The formulation can suitably be presented in unit dosage form and can be prepared by any method well known in the pharmaceutical arts. The amount of the active ingredient that can be combined with the carrier material to produce a single dosage form will vary depending on the host being treated and the particular mode of administration. The amount of the active ingredient that can be combined with the carrier material to produce a single dosage form will generally be the amount of the compound that produces a therapeutic effect. Generally, this amount will be in the range of about 1% to about 99%, preferably about 5% to about 70%, and most preferably about 10% to about 30% of the active ingredient, by weight.

[0180] The methods for preparing such preparations or compositions include the step of bringing the active compound (such as the compounds of the present invention) into association with a carrier and, optionally, one or more accessory ingredients. Generally, the preparations are prepared by uniformly and intimately bringing the compounds of the present invention into association with a liquid carrier or a finely divided solid carrier or both and, if necessary, then shaping the product.

[0181] Preparations of the present invention suitable for oral administration may be in the form of capsules (including dispersed capsules and gelatin capsules), cachets, pills, troches, lozenges (using a flavoring base, usually sucrose and acacia or tragacanth), lyophilizates, powders, granules, or in the form of solutions or suspensions in aqueous or non-aqueous liquids, or in the form of water-in-oil or oil-in-water liquid emulsions, or in the form of elixirs or syrups, or in the form of tablets (using an inert base, such as gelatin and glycerin, or sucrose and acacia), and / or in the form of mouthwashes and the like, each containing a predetermined amount of a compound of the present invention as an active ingredient. The compositions or compounds may also be administered in the form of boluses, pastilles, or poultices.

[0182] For preparing solid dosage forms for oral administration (capsules (including dispersed capsules and gelatin capsules), troches, pills, dragees, powders, granules, and the like), the active ingredient is mixed with one or more pharmaceutically acceptable carriers, such as sodium citrate or calcium phosphate, and / or any of the following: (1) fillers or extenders, such as starch, lactose, sucrose, glucose, mannitol, and / or silicic acid; (2) binders, such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and / or acacia; (3) humectants, such as glycerol; (4) disintegrants, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate; (5) solution retarders, such as paraffin; (6) absorption accelerators, such as quaternary ammonium compounds; (7) wetting agents, such as, for example, cetyl alcohol and glycerol monostearate; (8) adsorbents, such as kaolin and bentonite; (9) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof; (10) complexing agents, such as modified and unmodified cyclodextrins; and (11) coloring agents. In the case of capsules (including dispersed capsules and gelatin capsules), troches, and pills, the pharmaceutical compositions may also contain buffering agents. Solid compositions of a similar type may also be used as fillers in soft and hard gelatin capsules using excipients such as lactose (lactose / milk sugar) and high molecular weight polyethylene glycols and the like.

[0183] Tablets can be manufactured by compressing or molding, optionally with one or more accessory ingredients. Compressed tablets can be prepared using binders (such as gelatin or hydroxypropylmethylcellulose), lubricants, inert diluents, preservatives, disintegrants (such as sodium starch glycolate or croscarmellose sodium), surfactants or dispersing agents. Molded tablets can be manufactured by molding a mixture of the powdered compound moistened with an inert liquid diluent in a suitable machine.

[0184] Tablets and other solid dosage forms of the pharmaceutical composition, such as dragees, capsules (including dispersed capsules and gelatin capsules), pills and granules, can be prepared, optionally with scoring or using coatings and shells, such as enteric coatings and other coatings well-known in pharmaceutical compounding techniques. They can also be formulated using, for example, different proportions of hydroxypropylmethylcellulose (to provide the desired release profile), other polymeric matrices, liposomes and / or microspheres to provide slow or controlled release of the active ingredient therein. They can be sterilized by, for example, filtration through a bacteria-retaining filter or by incorporating a sterilizing agent in the form of a sterile solid composition that is soluble in sterile water or some other sterile injectable medium just before use. Such compositions can also optionally contain emulsifying agents and can be compositions that release the active ingredient solely or preferably in a delayed manner in a particular part of the gastrointestinal tract. Examples of embedding compositions that can be used include polymeric substances and waxes. The active ingredient can also be in microencapsulated form, if appropriate, together with one or more of the above excipients.

[0185] Liquid dosage forms suitable for oral administration include pharmaceutically acceptable emulsions, lyophilized products for reconstitution, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active ingredient, liquid dosage forms can contain inert diluents commonly used in the art, such as (for example) water or other solvents; cyclodextrins and their derivatives; solubilizing and emulsifying agents, such as ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butanediol, oils (specifically cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil and sesame oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol and fatty acid esters of sorbitan and mixtures thereof.

[0186] In addition to the inert diluent, oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening agents, flavoring agents, coloring agents, aromatic agents and preservatives.

[0187] In addition to the active compound, suspensions can also contain suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum hydroxide, bentonite, agar-agar and tragacanth and mixtures thereof.

[0188] Dosage forms for topical or percutaneous administration include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches, and inhalants. The active compounds can be mixed under sterile conditions with a pharmaceutically acceptable carrier and any preservatives, buffers, or propellants as may be required.

[0189] In addition to the active compound, ointments, pastes, creams, and gels may also contain excipients such as animal and vegetable fats, oils, waxes, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycols, polysiloxanes, bentonite, silicic acid, talc, and zinc oxide, or mixtures thereof.

[0190] In addition to the active compound, powders and sprays may also contain excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicate, and polyamide powder, or mixtures of such substances. Sprays may additionally contain conventional propellants such as chlorofluorocarbons and volatile unsubstituted hydrocarbons such as butane and propane.

[0191] Transdermal patches have the additional advantage of providing controlled delivery of the compounds of the present invention to the body. Such dosage forms can be manufactured by dissolving or dispersing the active compound in a suitable medium. Absorption enhancers can also be used to increase the amount of the compound permeating through the skin. The rate of such flux can be controlled by providing a rate-controlling membrane or by dispersing the compound in a polymer matrix or gel.

[0192] Examples of suitable aqueous and non-aqueous carriers that can be used in the pharmaceutical compositions of the present invention include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, and the like), and suitable mixtures thereof, vegetable oils (such as olive oil), and injectable organic esters (such as ethyl oleate). Appropriate fluidity can be maintained, for example, by using coating materials such as lecithin, by maintaining the desired particle size in the case of dispersions, and by using surfactants.

[0193] Such compositions may also contain adjuvants such as preservatives, wetting agents, emulsifying agents, and dispersing agents. Microbial action can be prevented by including various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, and the like. It may also be necessary to include isotonic agents such as sugars, sodium chloride, and the like in the compositions. In addition, extended absorption of injectable pharmaceutical forms can be achieved by including agents that delay absorption such as aluminum monostearate and gelatin.

[0194] In some cases, to prolong the action of a drug, it is necessary to slow down the absorption of the drug from subcutaneous or intramuscular injection. This can be achieved by using a liquid suspension of a crystalline or amorphous material with poor water solubility. The absorption rate of the drug depends on its dissolution rate, which in turn depends on the crystal size and crystalline form. Alternatively, delayed absorption of parenterally administered drug forms is achieved by dissolving or suspending the drug in an oily vehicle.

[0195] Injectable depot forms are made by forming a microencapsulation matrix of the compounds of the invention in a biodegradable polymer such as poly(lactide-co-glycolide). The rate of drug release can be controlled depending on the drug-to-polymer ratio and the nature of the particular polymer used. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Injectable depot formulations are also prepared by coating the drug in liposomes or microemulsions that are compatible with body tissues.

[0196] For use in the methods of the invention, the active compounds can be administered per se or in the form of pharmaceutical compositions containing, for example, from 0.1% to 99.5% (more preferably from 0.5% to 90%) of the active ingredient and a pharmaceutically acceptable carrier.

[0197] Introduction methods can also be provided by refillable or biodegradable devices. In recent years, various slow-release polymeric devices for the controlled delivery of drugs, including protein biopharmaceuticals, have been developed and tested in vivo. A variety of biocompatible polymers, including hydrogels, comprising biodegradable and non-biodegradable polymers can be used to form implants that continuously release the compound at a specific target site.

[0198] The actual dosage level of the active ingredient in the pharmaceutical compositions can vary so as to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration without being toxic to the patient.

[0199] The selected dosage level will depend on a variety of factors including the activity of the particular compound or combination of compounds or their esters, salts or amides used, the route of administration, the time of administration, the rate of excretion of the particular compound used, the duration of the treatment, other drugs, compounds and / or materials used in combination with the particular compound, the age, sex, weight, condition, general health and prior medical history of the patient being treated, and like factors well known in the medical arts.

[0200] A physician or veterinarian of ordinary skill in the art can readily determine and prescribe a therapeutically effective amount of the desired pharmaceutical composition. For example, the physician or veterinarian can begin administration of the pharmaceutical composition or compound at a level lower than that required to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved. "Therapeutically effective amount" means a concentration of the compound sufficient to elicit the desired therapeutic effect. It is generally understood that the effective amount of a compound will vary according to the weight, sex, age, and medical history of the subject. Other factors that can affect the effective amount can include (but are not limited to) the severity of the patient's condition, the disorder being treated, the stability of the compound, and another type of therapeutic agent to be co-administered with the compounds of the present invention as needed. A larger total dose can be delivered by administering the agent multiple times. Methods for determining efficacy and dosage are known to those of skill in the art (Isselbacher et al. (1996) Harrison’s Principles of Internal Medicine 13th ed., 1814-1882, incorporated herein by reference).

[0201] Generally, the suitable daily dose of the active compound used in the compositions and methods of the present invention should be the amount of the compound that is the lowest dose effective to produce a therapeutic effect. Such effective doses generally will depend on the factors described above.

[0202] When needed, the effective daily dose of the active compound can be administered in unit dosage form as one, two, three, four, five, six, or more sub-doses administered at appropriate time intervals throughout the day. In certain embodiments of the present invention, the active compound can be administered two or three times a day. In a preferred embodiment, the active compound will be administered once a day.

[0203] The patients to be treated are any animals in need, generally including primates (specifically humans); and other mammals such as horses, cattle, pigs, sheep, cats, and dogs; poultry; and pets.

[0204] In certain embodiments, the compounds of the present invention can be used alone or co-administered with another type of therapeutic agent. As used herein, the phrase "co-administered" refers to any form of administration of two or more different therapeutic compounds such that the second compound is administered while a previously administered therapeutic compound is still effective in the body (e.g., both compounds are effective simultaneously in the patient, which can include a synergistic effect of the two compounds). For example, the different therapeutic compounds can be administered simultaneously or sequentially in the same formulation or in separate formulations. In certain embodiments, the different therapeutic compounds can be administered to each other within one hour, 12 hours, 24 hours, 36 hours, 48 hours, 72 hours, or one week. Thus, a subject receiving such treatment can benefit from the combined action of the different therapeutic compounds.

[0205] In certain embodiments, co - administration of the compounds of the present invention with one or more other therapeutic agents (e.g., one or more other chemotherapeutic agents) provides improved efficacy relative to the separate administration of the compounds of the present invention or the one or more other therapeutic agents. In certain such embodiments, co - administration provides an additive effect, where an additive effect refers to the sum of the individual effects of separately administering the compounds of the present invention and the one or more other therapeutic agents.

[0206] In certain preferred embodiments, the compounds of the present invention are co - administered with a steroid anti - inflammatory drug, such as prednisolone, dexamethasone, prednisone, methylprednisolone, beclomethasone, betamethasone, fluticasone, or hydrocortisone. In certain preferred embodiments, the compounds of the present invention are co - administered with dexamethasone. Without wishing to be bound by any particular theory, it is believed that the steroid anti - inflammatory drugs produce a synergistic effect when used in combination with the compounds of the present invention, as described in U.S. Patent Application Publication No. 2018 / 0078567, which is hereby incorporated by reference in its entirety. The dosage forms of the compounds of the present invention and the steroid anti - inflammatory drugs can be appropriately modified by those skilled in the art. In certain preferred embodiments, the compounds of the present invention are co - administered with a cell cycle inhibitor. Cell cycle inhibitors suitable for use with the compounds of the present invention include cell cycle inhibitors known in the art, including (but not limited to) those described by Mills et al., “Recent Advances of Cell - Cycle Inhibitor Therapies for Pediatric Cancer”, Cancer Research (2017) doi:10.158 / 0008 - 5472.CAN - 17 - 2066. Exemplary cell cycle inhibitors suitable for use with the compounds of the present invention include (but not limited to) palbociclib, ribociclib, abemaciclib, flavopiridol, AT9283, Alisertib, and MK - 1775. The dosage forms of the compounds of the present invention and the cell cycle inhibitors can be suitably prepared by those skilled in the art.

[0207] In certain preferred embodiments, the compounds of the present invention are administered in combination with one or more other anti-apoptotic agents, such as Mcl-1 inhibitors and inhibitor of apoptosis protein (IAP) antagonists. Suitable Mcl-1 inhibitors include, but are not limited to, the Mcl-1 inhibitors described in Chen et al., "Mcl-1 Inhibitors: A Patent Review", Expert Opinion on Therapeutic Patents (2016) doi:10.1080 / 13543776.2017.1249848. Such compounds include, but are not limited to

[0208]

[0209]

[0210] Suitable IAP antagonists include, but are not limited to, the IAP antagonists described in Hird et al., "Small Molecule Inhibitor of Apoptosis Proteins Antagonists: a Patent Review", Expert Opinion on Therapeutic Patents (2015) doi:10.1517 / 13543776.2015.1041922, which is incorporated herein by reference in its entirety. Such compounds include, but are not limited to

[0211]

[0212]

[0213]

[0214]

[0215]

[0216] AT-406 / Debio1143, GDC-0917 / CUDC-427, LCL161 and TL-32711.

[0217] Wetting agents, emulsifying agents and lubricants (such as sodium lauryl sulfate and magnesium stearate) as well as coloring agents, release agents, coating agents, sweetening agents, flavoring agents and fragrances, preservatives and antioxidants may also be present in the composition.

[0218] Examples of pharmaceutically acceptable antioxidants include: (1) water-soluble antioxidants such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite and the like; (2) oil-soluble antioxidants such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, alpha-tocopherol and the like; and (3) metal chelating agents such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid and the like.

[0219] The compound represented by formula (I), its salt or its prodrug can inhibit the growth of leukemia stem cells or kill leukemia stem cells. Therefore, such a compound, its salt or its prodrug can be used in a pharmaceutical composition for treating acute myeloid leukemia or for inhibiting the recurrence of acute myeloid leukemia.

[0220] Leukemia stem cells refer to cells that meet at least one of the following requirements:

[0221] 1: It is a cell that can selectively and exclusively manifest leukemia in vivo;

[0222] 2: It is a cell that can generate a leukemia non-stem cell part that cannot spontaneously manifest leukemia;

[0223] 3: It is a cell that can be implanted in vivo; and / or

[0224] 4: It is a cell that can self-renew.

[0225] In certain embodiments, leukemia stem cells exhibit the characteristics of CD34 + CD38 - as surface antigens. In this specification, leukemia stem cells obtained from a patient with acute myeloid leukemia are also referred to as "human AML CD34 + CD38 - cells".

[0226] When a cell can self-renew, such a cell can divide into two cells; that is, one stem cell and more differentiated progenitor cells. The concept of leukemia stem cells has been established and is commonly recognized in this technology (D. Bonnet, J. E. Dick, Nat. Med., 3, 730 (1997); T. Lapidot et al., Nature, 367, 645 (1994)).

[0227] In this specification, all types of leukemia stem cells are within the scope of "leukemia stem cells". The term "leukemia stem cell" preferably refers to a stem cell with a high expression level of the HCK gene, and more preferably refers to a stem cell of acute myeloid leukemia cells.

[0228] The target leukemia stem cells in the present invention generally originate from mammals. Examples of mammals include: laboratory animals such as mice, rats, hamsters, guinea pigs and other rodents, and rabbits; domestic animals such as pigs, cows, goats, horses, sheep and minks; companion animals such as dogs and cats; and primates such as humans, monkeys, cynomolgus monkeys, rhesus monkeys, marmosets, orangutans and chimpanzees. The target leukemia stem cells in the present invention preferably originate from primates (such as humans) or rodents (such as mice).

[0229] The pharmaceutical composition for killing leukemia stem cells and the pharmaceutical composition for treating acute myeloid leukemia or inhibiting the recurrence of acute myeloid leukemia according to the present invention (hereinafter "the pharmaceutical composition of the present invention") have the effect of killing leukemia stem cells. The effect of killing leukemia stem cells can be confirmed according to the effect of inhibiting the growth of leukemia stem cells in the whole cell population.

[0230] Since leukemia stem cells are regarded as the cause of recurrent leukemia, the use of the pharmaceutical composition of the present invention can inhibit and / or prevent the recurrence of leukemia. Specifically, the pharmaceutical composition of the present invention can also be used to contain leukemia (preferably used as an agent for inhibiting the recurrence of leukemia). Leukemia recurrence is a situation where leukemia cells grow again and leukemia symptoms reappear or worsen after the patient has achieved partial or complete remission of leukemia symptoms by treatment. By administering the pharmaceutical composition of the present invention to a mammal at risk of developing leukemia (or experiencing a recurrence of leukemia), the development of leukemia (or the recurrence of leukemia) can be inhibited and / or prevented.

[0231] Acute myeloid leukemia can be completely remitted by using known chemotherapeutic agents for cancer, such as alkylating agents (e.g., cyclophosphamide and ifosfamide), antimetabolites (e.g., cytarabine, 5-fluorouracil and methotrexate), antitumor antibiotics (e.g., adriamycin and mitomycin), anticancer agents of plant origin (e.g., vinblastine, vincristine, vindesine and Taxol), cisplatin, carboplatin or etoposide. However, AML usually recurs after complete remission has been achieved, and recurrent AML leads to death in many patients.

[0232] The recurrence of acute myeloid leukemia can be prevented by administering the pharmaceutical composition of the present invention to patients in remission of acute myeloid leukemia.

[0233] The compound represented by formula (I) inhibits HCK and / or FLT3 and can retain inhibitor activity in cells with FLT3 / ITD mutations. Therefore, a pharmaceutical composition containing the compound, its salt or its prodrug of the present invention may be suitable for killing leukemia stem cells. The pharmaceutical composition containing the compound, its salt or its prodrug of the present invention can also kill leukemia stem cells with FLT3 / ITD mutations. Therefore, such a pharmaceutical composition can treat acute myeloid leukemia, including acute myeloid leukemia caused by leukemia stem cells with one or more FLT3 / ITD mutations.

[0234] In addition to the compound, its salt or its prodrug represented by formula (I), the pharmaceutical composition of the present invention may further contain an agent (e.g.) that exerts an inhibitory effect on FLT3 to enhance the efficacy against leukemia stem cells (including leukemia stem cells with Flt3 / ITD mutations).

[0235] Examples of the agent having an inhibitory effect on FLT3 used in the present invention include (but are not limited to) Crenolanib, lestaurtinib (CEP-701 / KT5555), PKC412 (CGP41251), tandutinib (MLN518 / CT53518), sunitinib (SU11248), sorafenib (BA43-9006), linifanib (ABT-869), dovitinib (CHIR-258 / TKI-258), KW-2449, quizartinib (AC220), dovitinib dilactate, cabozantinib (XL-184), R406, TG101209, amuvatinib, and ENMD-2076.

[0236] The ITD mutation in FLT3 can be detected and evaluated by detecting the difference from the normal type at the mutation site as needed, by PCR, electrophoresis, sequencing, detection with an antibody (e.g., Western blotting or ELISA), or other means. It can be evaluated that the pharmacological efficacy of a subject in whom an ITD mutation in FLT3 is detected in leukemia stem cells is enhanced compared to a subject in whom normal FLT3 is detected in leukemia stem cells. When the pharmacological efficacy is evaluated as enhanced, the pharmaceutical composition of the present invention is administered to the subject. Therefore, the treatment, inhibition, and / or prevention of the recurrence of acute myeloid leukemia can be achieved.

[0237] Application

[0238] The compositions of the present invention can be administered orally, parenterally (including subcutaneously, intramuscularly, intravenously and intradermally), by inhalation spray, topically, rectally, nasally, buccally, vaginally or via an implantable reservoir. In some embodiments, the provided compounds or compositions can be administered intravenously and / or intraperitoneally. In certain preferred embodiments, the disclosed methods include orally or parenterally administering any two or all three of an FLT3-ITD inhibitor, an HCK inhibitor, and a BCL-2 inhibitor.

[0239] As used herein, the term "parenteral" includes subcutaneous, intravenous, intramuscular, intraocular, intravitreal, intra-articular, intrasynovial, intrasternal, intrathecal, intrahepatic, intraperitoneal, intralesional and intracranial injection or infusion techniques. Preferably, the compositions are administered orally, subcutaneously, intraperitoneally or intravenously. The sterile injectable form of the compositions of the present invention can be an aqueous or oily suspension. Such suspensions can be formulated using suitable dispersing or wetting agents and suspending agents according to techniques known in the art. The sterile injectable preparation can also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, such as in the form of a solution in 1,3-butanediol. Among the acceptable solvents and diluents that can be employed are water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium.

[0240] The pharmaceutically acceptable compositions of the present invention can be administered orally in any acceptable oral dosage form, including (but not limited to) capsules, lozenges, aqueous suspensions or solutions. In the case of lozenges for oral use, the carriers commonly used include lactose and corn starch. Lubricants, such as magnesium stearate, are also usually added. For oral administration in the form of capsules, suitable diluents include lactose and anhydrous corn starch. When an aqueous suspension is required for oral use, the active ingredient is combined with emulsifying and suspending agents. If desired, certain sweetening, flavoring or coloring agents can also be added. In some embodiments, the provided oral formulations are formulated for immediate release or sustained / delayed release. In some embodiments, the compositions are suitable for buccal or sublingual administration, including lozenges, troches and tablets. The compounds disclosed herein can also be in microencapsulated form.

[0241] The amount of the compounds of the invention that can be combined with a carrier material to produce a composition in a single dosage form will vary depending on the subject being treated and the particular mode of administration. In certain embodiments, the provided compositions should be formulated such that a dose between about 0.01 and about 100 mg / kg body weight / day of the compound can be administered to a subject receiving such a composition. In other embodiments, the dose is about 0.5 to about 100 mg / kg body weight every about 4 to 120 hours or between about 1 mg and about 1000 mg / dose, or according to the requirements of the particular drug. Generally, the pharmaceutical compositions of the invention should be administered about 1 to about 6 times per day.

[0242] In some embodiments, the compound is formulated for oral administration at a dose of from approximately 5 mg / kg to approximately 10 mg / kg, preferably at a dose of approximately 7.5 mg / kg.

[0243] It should also be understood that the specific dosage and treatment regimen for any particular subject will depend on a variety of factors, including the activity of the specific compound used, age, body weight, general health, sex, diet, time of administration, rate of excretion, drug combination, and the judgment of the treating physician, as well as the severity of the particular disease being treated. The amount of the compounds of the invention in the composition will also depend on the particular compound in the composition.

[0244] When the condition of the subject improves, a maintenance dose of the compounds, compositions, or combinations of the invention may be administered if needed. Subsequently, the dosage or frequency or both may be reduced according to the symptoms to a level that maintains the improvement of the condition when the symptoms have been alleviated to the desired level. However, the subject may require long-term intermittent treatment upon recurrence of any disease symptoms.

[0245] Incorporation of References

[0246] All publications, patents, and patent applications mentioned in this specification are incorporated herein by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference. In case of conflict, the present application (including any definitions herein) shall govern. Examples

[0247] Using no more than routine experimentation, those skilled in the art will recognize or be able to ascertain many equivalents to the specific procedures, embodiments, techniques, and examples described herein. Such equivalents are considered to be within the scope of the invention and are covered by the patent claims appended hereto. For example, it should be understood that modifying reaction conditions with alternatives recognized in the art and using no more than routine experimentation are within the scope of this application.

[0248] The following examples are presented to provide a complete disclosure and description of how to make and use the method of the present invention to those of ordinary skill in the art, and are not intended to limit the scope which the inventors regard as their invention.

[0249] Unless otherwise mentioned, the starting materials used in the experiments described herein were obtained from commercial sources or known procedures and used without further modification.

[0250] General method

[0251] Compound

[0252] General method

[0253] Unless otherwise indicated, all solvents and reagents were obtained from commercial sources and used without further purification. NMR spectra were obtained on a Bruker Neo 400M spectrometer operating at 400 MHz. Chemical shifts were reported in parts per million (δ) relative to the tetramethylsilane resonance in the specified solvent. LC-MS was obtained using an Agilent 1260-6125B single quadrupole mass spectrometer with a Welch Biomate column (C18, 2.7um, 4.6×50mm) eluted with a mixture of solvent A (ACN containing 0.05% FA) and B (water containing 0.05% FA) using gradient elution. Detection was performed by DAD (254 nm and 210 nm). Ionization was performed by ESI. Spectra were analyzed using Chemstation software. Analytical HPLC was performed on a Waters ARC system under acidic conditions on a YMC Pack Pro column (C18 S-3um, 12nm, 150×2.0mm) eluted with a mixture of solvent A (ACN containing 0.05% FA) and B (water containing 0.05% FA); or under basic conditions on an Agilent Poroshell HPH C18 column (2.7um, 2.1×150mm) eluted with a mixture of solvent C (water containing 0.1% NH 4 OH) and D (ACN containing 0.1% NH 4 OH) using gradient elution. Detection was performed by DAD (254 nm and 210 nm). Preparative HPLC was performed on a Waters AutoP system combined with a single quadrupole mass spectrometer using a Welch C18 column (5um, 25×150mm) eluted with a mixture of solvent A and B. Flash chromatography was performed on a Biotage Isolera Prime system using a Welch WelFlash flash column (40-63um) eluted with a solvent mixture as specified in the experimental procedure.

[0254] 4-(4-Amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexanone

[0255]

[0256] 4-Chloro-5-iodo-7-(1,4-dioxaspiro[4.5]dec-8-yl)-7H-pyrrolo[2,3-d]pyrimidine

[0257] DEAD (170 mL, 1.07 mol) was added dropwise to a stirred mixture of Ph 3 P (285.1 g, 1.07 mol) and THF (3.12 L) under ice-bath cooling. After the reaction, the mixture was warmed to room temperature. A solution of 4-dichloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidine (100 g, 0.36 mol) and 1,4-dioxaspiro[4.5]dec-8-ol (170 g, 1.07 mol) in THF (1.2 L) was added dropwise over a 60-minute period. The reaction mixture was stirred at room temperature for 3 h. TLC (EA:PE = 1:4) showed complete depletion of the starting materials. The solvent was evaporated in vacuo. EA (1.78 L) was added to the residue and the solid was collected by filtration. The solid was washed with EA (300 mL) and dried under reduced pressure to give the title compound as a white solid (139.8 g, 92.7% yield). LCMS: calculated exact mass = 419.0, experimental value [M+H] + (ESI) = 420.0; 1 H NMR (DMSO-d 6 ) δ ppm: 8.64 (s, 1H), 8.12 (s, 1H), 4.70 - 4.80 (m, 1H), 3.88 - 3.95 (m, 4H), 2.08 - 2.17 (m, 2H), 1.88 - 1.96 (m, 2H), 1.70 - 1.84 (m, 4H).

[0258] 4-Dichloro-5-(4-phenoxyphenyl)-7-(1,4-dioxaspiro[4.5]dec-8-yl)-7H-pyrrolo[2,3-d]pyrimidine

[0259] 4-Dichloro-5-iodo-7-(1,4-dioxaspiro[4.5]dec-8-yl)-7H-pyrrolo[2,3-d]pyrimidine (60 g, 143 mmol), (4-phenoxyphenyl)boronic acid (60 g, 286 mmol), Pd(dppf)Cl 2 (10 g, 14.3 mmol) and Na 2 CO 3(45 g, 429 mmol) in dioxane-H 2 O (10:1) (2.5 L) was heated to 80 °C and stirred for 3 h. TLC (PE:EA = 4:1) showed complete depletion of the starting material. The solvent was evaporated in vacuo. The residue was extracted with DCM (500 mL × 2). The combined organic layers were washed with water, dried over Na 2 SO 4 and concentrated in vacuo. The crude product was purified by flash column chromatography (EA:PE = 1:4) to give the title compound as a brown solid (42.0 g, 66% yield). LCMS: calculated exact mass = 461.1, found [M+H] + (ESI) = 462.2; 1 H NMR (DMSO-d 6 ) δ ppm: 8.66 (s, 1H), 7.95 (s, 1H), 7.54 - 7.56 (m, 2H), 7.41 - 7.45 (m, 2H), 7.03 - 7.21 (m, 5H), 4.78 - 4.87 (m, 1H), 3.89 - 3.96 (m, 4H), 2.12 - 2.26 (m, 2H), 1.94 - 2.02 (m, 2H), 1.74 - 1.88 (m, 4H).

[0260] 5-(4-Phenoxyphenyl)-7-(1,4-dioxaspiro[4.5]dec-8-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0261] A reaction mixture of 4-dichloro-5-(4-phenoxyphenyl)-7-(1,4-dioxaspiro[4.5]dec-8-yl)-7H-pyrrolo[2,3-d]pyrimidine (20.3 g, 43.9 mmol), NH 4 OH (200 mL) and dioxane (200 mL) was stirred in a high-pressure reactor at 120 °C for 48 h. The solid was filtered and dried to afford the title compound as a white solid (10 g, 51.5% yield). LCMS: calculated exact mass = 442.2, found [M+H] + (ESI) = 443.2.

[0262] 4-(4-Amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexanone

[0263] A reaction mixture of 5-(4-phenoxyphenyl)-7-(1,4-dioxaspiro[4.5]dec-8-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (10 g, 22.6 mmol), HCl (6 N, 160 mL), acetone (160 mL), and THF (36 mL) was stirred at 40 °C for 2 h. It was cooled in an ice bath, and NaOH (1 M, 980 mL) was added to the mixture until pH 8. The solid was collected by filtration and dried under reduced pressure to give the title compound as a white solid (7.6 g, 84.4% yield). LCMS: calculated exact mass = 398.2, experimental value [M+H] + (ESI) = 399.2; 1 H NMR (DMSO-d 6 ) δ ppm: 8.17 (s, 1H), 7.40 - 7.47 (m, 5H), 7.08 - 7.17 (m, 5H), 6.12 (br.s., 2H), 5.18 (s, 1H), 2.73 - 2.78 (m, 2H), 2.28 - 2.40 (m, 4H), 2.15 - 2.25 (m, 2H).

[0264] 5-Iodo-7-(1,4-dioxaspiro[4.5]dec-8-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0265]

[0266] 5-Iodo-7-(1,4-dioxaspiro[4.5]dec-8-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0267] A mixture of 4-chloro-5-iodo-7-(1,4-dioxaspiro[4.5]dec-8-yl)-7H-pyrrolo[2,3-d]pyrimidine (13 g, 31 mmol), NH 4 OH (200 mL), and dioxane (200 mL) was stirred in an autoclave at 120 °C for 48 h. TLC (MeOH:DCM = 1:20) showed complete depletion of the starting material. The title compound as a white solid was collected by filtration followed by drying under reduced pressure (11.6 g, 93% yield). LCMS: calculated exact mass = 400.0, experimental value [M+H] + (ESI+) = 401.0.

[0268] 5-Iodo-7-(1,4-dioxaspiro[4.5]dec-8-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0269] A mixture of 5-iodo-7-(1,4-dioxaspiro[4.5]dec-8-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (11.6 g, 29 mmol), 6 M HCl (160 mL), acetone (160 mL), and THF (38 mL) was stirred at 40 °C for 2 h. TLC (EA:PE = 1:3) showed complete depletion of the starting material. NaOH solution (980 mL, 1 mol / L) was added to the mixture until pH 8.0 (under ice-cooling). The title compound as a white solid was collected by filtration and then dried under reduced pressure (7.6 g, 73% yield). LCMS: calculated exact mass = 356.0, experimental value [M+H] + (ESI) = 357.0; 1 H NMR (DMSO-d 6 ) δ ppm: 8.12 (s, 1H), 7.60 (s, 1H), 6.65 (br.s., 2H), 5.10 (s, 1H), 2.68 - 2.74 (m, 2H), 2.23 - 2.30 (m, 4H), 2.09 - 2.14 (m, 2H).

[0270] 4-(4-Amino-5-(2-fluoro-4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexanone

[0271]

[0272] 4-(4-Amino-5-(2-fluoro-4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexanone

[0273] A solution of 4-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexanone (300 mg, 0.84 mmol), 2-(2-fluoro-4-phenoxyphenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (344.0 mg, 1.10 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (123.3 mg, 0.17 mmol), and sodium carbonate (178.6 mg, 1.65 mmol) in dioxane-water (11 mL, 10:1) was stirred at 90 °C under nitrogen for 12 h. After cooling to room temperature, the mixture was filtered through Celite and purified by concentration via flash chromatography (DCM:MeOH = 20:1) to afford a brown oil (400 mg, 114.6% yield, ~80% purity). LCMS: calculated exact mass = 416.2; experimental value [M+H]+ (ESI) = 417.0.

[0274] 4-(4-Amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-1-yl)cyclohexanone

[0275]

[0276] 3-Iodo-1H-pyrazolo[3,4-d]pyrimidin-4-amine

[0277] NIS (1.99 kg, 8.88 mol) was added in three portions to a mixture of 3-iodo-1H-pyrazolo[3,4-d]pyrimidin-4-amine (1.0 kg, 7.4 mol) in DMF (6 L). The reaction mixture was stirred overnight at 80 °C. The reaction mixture was cooled to room temperature and precipitated. The solid was filtered and washed with EtOAc (12 L). The solid was filtered and dried in vacuo to give the product (1.58 kg) as a white solid. LCMS: calculated exact mass = 260.9; experimental value [M+H] + (ESI) = 262.2; 1 HNMR(DMSO-d 6 ) δ ppm: 8.18 (s, 1H).

[0278] 3-Iodo-1-(1,4-dioxaspiro[4.5]dec-8-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine

[0279] To a suspension of 3-iodo-1H-pyrazolo[3,4-d]pyrimidin-4-amine (400 g, 1.53 mol) in THF (5.5 L) cooled in an ice bath was added 1,4-dioxaspiro[4.5]dec-8-ol (480 g, 3.06 mol), PPh 3 (600 g, 2.3 mol), followed by dropwise addition of DEAD (400 g, 3 mol) over a period of 4 h. The reaction mixture was stirred at room temperature for 1 h and monitored by LCMS until complete conversion of the starting material. The reaction mixture was concentrated. The crude product was washed with THF (400 mL) and then with EtOAc (4 L) to give the product (380 g, 62% yield). LCMS: calculated exact mass = 401.0; experimental value [M+H] + (ESI) = 401.9; 1 H NMR (400 MHz, DMSO-d 6 ) δ ppm: 8.19 (s, 1H) 4.64 - 4.79 (m, 1H) 3.85 - 3.96 (m, 4H) 2.09 - 2.23 (m, 2H) 1.66 - 1.90 (m, 6H).

[0280] 4-(4-Amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-1-yl)cyclohexanone

[0281] To a suspension of 3-iodo-1-(1,4-dioxaspiro[4.5]decan-8-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (660 g, 1.64 mol) in THF (1.2 L) and acetone (6 L) was added 6N HCl (6 L, 36 mol). The reaction mixture was heated at 60 °C for 1 hour. It was neutralized with 7.5N aqueous NaOH. After filtration, the solid product (350 g) was collected and used without further purification. LCMS: calculated exact mass = 357.0; experimental value [M+H] + (ESI) = 357.8; 1 H NMR (400 MHz, DMSO-d 6 ) δ ppm: 8.23 (s, 1H) 5.10 - 5.23 (m, 1H) 2.61 - 2.76 (m, 2H) 2.25 - 2.40 (m, 5H) 2.14 - 2.23 (m, 2H).

[0282] 4-(4-Amino-3-(2-fluoro-4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)cyclohexan-1-one

[0283]

[0284] 4-(4-Amino-3-(2-fluoro-4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)cyclohexan-1-one

[0285] To a 50 mL round-bottom flask were added 4-(4-amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-1-yl)cyclohexanone (3 g, 8.4 mmol), 2-(2-fluoro-4-phenoxyphenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (3.16 g, 10.08 mmol), Pd(dppf)Cl 2 (614 mg, 0.84 mmol), Na 2 CO 3 (3.6 g, 33.5 mmol), 30 mL of dioxane-water (9 - 1, 20 mL). The reaction mixture was stirred overnight at 85 °C under N 2 atmosphere. The reaction mixture was cooled to room temperature, filtered and concentrated. The crude product was purified by flash column chromatography (MeOH / DCM, 0% - 5%) to give the product as a red solid (2.5 g, 71.3% yield), which was used without further purification. LC-MS: calculated exact mass: 417.16; experimental value [M+H] + (ESI) = 418.15.

[0286] 5-Iodo-7-((cis)-4-(4-methylpiperazin-1-yl)cyclohexyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0287] and

[0288] 5-Iodo-7-((trans)-4-(4-methylpiperazin-1-yl)cyclohexyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0289]

[0290] A mixture of 5-iodo-7-(1,4-dioxaspiro[4.5]dec-8-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (6.3 g, 17.7 mmol), 1-methylpiperazine (11.71 g, 106.2 mmol), and AcOH (0.63 mL, 10.62 mmol) in DCM (50 mL) was stirred at room temperature for 16 h. NaCNBH 3 (2.21 g, 35.4 mmol) was added to the mixture. The mixture was then stirred at 50 °C for 4 h. TLC (MeOH:DCM = 1:15) showed complete depletion of the starting material. The solvent was evaporated in vacuo. The residue was extracted with DCM (250 mL × 2). The combined organic layers were washed with water, dried over anhydrous Na 2 SO 4 and concentrated in vacuo. The residue was purified by flash column chromatography (DCM:MeOH = 10:1) to give 5-iodo-7-((trans)-4-(4-methylpiperazin-1-yl)cyclohexyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (4.3 g, 55.3% yield) and the cis isomer. Analytical data for the trans isomer: LCMS: calculated exact mass = 440.1, experimental value [M+H] + (ESI) = 441.1; 1 HNMR (DMSO-d 6 ) δ ppm: 8.08 (s, 1H), 7.53 (s, 1H), 6.58 (br.s., 2H), 4.47 (s, 1H), 2.35 (br.s., 4H), 2.16 (s, 3H), 1.81 - 1.90 (m, 6H), 1.38 - 1.44 (m, 2H).

[0291] 5-Iodo-7-((cis)-4-(4-methylpiperazin-1-yl)cyclohexyl)-7H-pyrazolo[2,3-d]pyrimidin-4-amine

[0292] and

[0293] 5-Iodo-7-((trans)-4-(4-methylpiperazin-1-yl)cyclohexyl)-7H-pyrazolo[2,3-d]pyrimidin-4-amine

[0294]

[0295] 5-Iodo-7-((cis)-4-(4-methylpiperazin-1-yl)cyclohexyl)-7H-pyrazolo[2,3-d]pyrimidin-4-amine

[0296] A solution of 4-(4-amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-1-yl)cyclohexanone (700 mg, 1.96 mmol), 1-methylpiperazine (0.65 mL, 11.76 mmol), acetic acid (0.1 mL, 1.96 mmol) and molecular sieves (2.0 g) in 1,2-dichloroethane (40 mL) was stirred at room temperature for 1 hour. Subsequently, sodium triacetoxyborohydride (1.2 g, 11.76 mmol) was added and stirring was continued at room temperature for 16 hours. Water (150 mL) and dichloromethane (150 mL) were added to the reaction mixture, a saturated aqueous solution of sodium bicarbonate (120 mL) was further added thereto and the mixture was partitioned. The organic layer was washed with brine (120 mL), dried over sodium sulfate and concentrated by flash chromatography (DCM:MeOH = 10:1) to purification to obtain 5-iodo-7-((trans)-4-(4-methylpiperazin-1-yl)cyclohexyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (530 mg, 61.6% yield) as a white solid and the corresponding cis isomer. Analytical data for the trans isomer: LCMS: calculated exact mass = 440.1; experimental value [M+H] + (ESI) = 442.1.

[0297] (S)-tert-Butyl 4-((trans)-4-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexyl)-2-methylpiperazine-1-carboxylate

[0298] and

[0299] (S)-tert-Butyl 4-((cis)-4-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexyl)-2-methylpiperazine-1-carboxylate

[0300]

[0301] At room temperature in N 2Under an atmosphere, NaBH 3 CN (1 g, 16.8 mmol) was added to a solution of 4-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexanone (1 g, 2.8 mmol), (S)-tert-butyl 2-methylpiperazine-1-carboxylate (3.36 g, 16.8 mmol), and AcOH (0.1 mL) in DCM (70 mL). Subsequently, the reaction mixture was stirred at 40 °C for 16 h. The reaction mixture was extracted with DCM (150 mL) and water. The combined organic layers were washed with an aqueous solution of NH 4 Cl (150 mL), dried over anhydrous Na 2 SO 4 and concentrated in vacuo. The residue was purified by flash column chromatography (DCM:MeOH = 20:1) to afford a mixture of the two title compounds (2 g), which was used without further purification. LCMS: calculated exact mass = 540.17, found [M+H] + (ESI) = 541.1.

[0302] 5-Iodo-7-((trans)-4-((S)-3-methylpiperazin-1-yl)cyclohexyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0303]

[0304] TFA (10 mL) was added dropwise to a solution of (S)-tert-butyl 4-((trans)-4-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexyl)-2-methylpiperazine-1-carboxylate (1.0 g, 1.85 mmol) in DCM (20 mL). The reaction mixture was stirred at room temperature for 0.5 h. The mixture was evaporated and purified by flash column chromatography (MeOH / DCM, 1% to 10% gradient) to afford the title compound as a pale solid (690 mg, 85% yield).

[0305] (S)-tert-Butyl 4-((trans)-4-(4-amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-1-yl)cyclohexyl)-2-methylpiperazine-1-carboxylate

[0306] and

[0307] (S)-tert-Butyl 4-((cis)-4-(4-amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-1-yl)cyclohexyl)-2-methylpiperazine-1-carboxylate

[0308]

[0309] A mixture of 4-(4-amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-1-yl)cyclohexanone (2.5 g, 7 mmol) and tert-butyl (S)-2-methylpiperazine-1-carboxylate (4.2 g, 21 mmol) in DCE (50 mL) was stirred at 55 °C under N 2 atmosphere for 90 min. After cooling it to room temperature, NaBH(OAc) 3 (2076 mg, 9.8 mmol) was added portionwise, and the mixture was stirred at 55 °C under N 2 atmosphere for 90 min, then cooled to room temperature and stirred overnight at room temperature. The reaction was monitored by TLC and LCMS until the starting materials were completely exhausted. Then it was filtered and concentrated to give the crude product. The crude product was purified by flash column chromatography (0 - 30% MeOH / DCM) to obtain the title compound as a yellow solid (1.4 g, 36% yield). LCMS: calculated exact mass = 541.17; experimental value [M+H] + (ESI) = 541.82.

[0310] 5-Iodo-7-((trans)-2-(1-methylpiperidin-4-yl)-1,3-dioxolan-5-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine and 5-iodo-7-((cis)-2-(1-methylpiperidin-4-yl)-1,3-dioxolan-5-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0311]

[0312] 2-Oxopropane-1,3-diyl dibenzoate

[0313] To a stirred solution of 1,3-dihydroxypropan-2-one (30 g, 333 mmol), DMAP (2.1 g, 16.65 mmol) in pyridine (133 mL) cooled in an ice bath, benzoyl chloride (85 mL, 732 mmol) was added dropwise, and then pyridine (120 mL) was added. After the addition, the mixture was warmed to room temperature and stirred overnight. The mixture was poured into HCl (6 M, 580 mL) at 0 °C. The mixture was filtered and washed with water (150 mL × 2). The residue was suspended in MeOH (400 mL), filtered to obtain the title compound as a white solid (64.6 g, 65% yield).

[0314] 2-Hydroxypropane-1,3-diyl dibenzoate

[0315] To a stirred solution of 2-oxopropane-1,3-diyl dibenzoate (15 g, 50 mmol) in THF (150 mL) cooled in an ice bath, NaBH4 (2.3 g, 60 mmol). After addition, the mixture was stirred at 0 °C for 30 minutes. The reaction mixture was quenched with H 4 Cl solution and evaporated. The residue was diluted with EA (200 mL), washed with water (80 mL) and brine (80 mL), dried and concentrated in vacuo and purified by flash column chromatography (EA / PE, 0 to 20% gradient) to afford the title compound as a colorless oil (9.02 g, 60% yield).

[0316] 2-(4-Chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)propane-1,3-diyl dibenzoate

[0317] To a solution of PPh 3 (15.5 g, 59.04 mmol) in THF (100 mL) cooled in an ice bath was added DEAD (10.3 g, 59.04 mmol). The mixture was stirred at room temperature. To the mixture was added a solution of 2-hydroxypropane-1,3-diyl dibenzoate (7.7 g, 25.6 mmol) and 4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidine (5.5 g, 19.68 mmol) in THF (60 mL). The reaction mixture was stirred at room temperature for 2 h. The mixture was evaporated. The residue was diluted with EA (30 mL), filtered and the filtrate was concentrated in vacuo and purified by flash column chromatography (EA / PE, 0 to 40% gradient) to afford the desired product as a white solid (7.1 g, 64% yield). LC-MS: Calculated exact mass: 561.0 Experimental value: [M+H] + (ESI) = 561.9; 1 H NMR (400 MHz, chloroform-d) δ ppm: 8.61 (s, 1H), 7.90 (m, 4H), 7.52 - 7.66 (m, 3H), 7.37 - 7.48 (m, 4H), 5.66 (m, 1H), 4.77 - 4.99 (m, 4H).

[0318] 2-(4-Amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)propane-1,3-diol

[0319] A solution of 2-(4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)propane-1,3-diyl dibenzoate (4.0 g, 7.12 mmol) in dioxane (50 mL) and concentrated aqueous ammonia solution (50 mL) were heated in a pressure vessel at 120 °C for 18 h. The mixture was cooled to room temperature and the solvent was distilled off in vacuo to give a yellow solid. The yellow solid was suspended in water (15 mL), filtered and washed with water (5 mL) to give a white solid. The white solid was suspended in MeOH (15 mL), filtered and washed with MeOH (5 mL) to give a white solid (2.0 g, 80% yield). LC-MS: Calculated exact mass: 334.0 Experimental value: [M+H] + (ESI) = 334.5.

[0320] 5-Iodo-7-((trans)-2-(piperidin-4-yl)-1,3-dioxan-5-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0321] and

[0322] 5-Iodo-7-((cis)-2-(piperidin-4-yl)-1,3-dioxan-5-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0323] A solution of 2-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)propane-1,3-diol (1.9 g, 5.69 mmol), tert-butyl 4-formylpiperidine-1-carboxylate (15.8 g, 73.93 mmol), TsOH-H 2 O (14.1 g, 73.93 mmol) and Na 2 SO 4 (40.4 g, 284.34 mmol) in toluene (250 mL) was stirred at 110 °C for 18 h. After cooling to room temperature, Na 2 CO 3 solution was added and stirred for 15 min. The mixture was concentrated. The solid was suspended in DCM / MeOH (v / v = 1:10, 400 mL), filtered, evaporated and purified by flash column chromatography (MeOH / DCM, 0 to 10% gradient) to give a yellow solid (880 mg, 36% yield). LC-MS: Calculated exact mass: 429.1 Experimental value: [M+H] + (ESI) = 429.6.

[0324] 5-Iodo-7-((trans)-2-(1-methylpiperidin-4-yl)-1,3-dioxan-5-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0325] and

[0326] 5-Iodo-7-((cis)-2-(1-methylpiperidin-4-yl)-1,3-dioxan-5-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0327] A solution of 5-iodo-7-((2r,5r)-2-(piperidin-4-yl)-1,3-dioxan-5-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (880 mg, 2.05 mmol), formaldehyde (369 mg, 12.3 mmol), and acetic acid (0.5 mL) in DCM / MeOH (v / v = 1:2, 45 mL) was stirred at room temperature for 1 hour. NaBH(OAc) 3 (3.9 g, 18.45 mmol) was added to the mixture. After stirring at room temperature for 16 hours, water (100 mL) and DCM (400 mL) were added to the mixture followed by saturated NaHCO 3 solution (200 mL). The organic layer was washed with brine (80 mL), dried over Na 2 SO 4 and evaporated and purified by flash column chromatography (MeOH / DCM, 0 to 10% gradient) to give a mixture of the title compound as a yellow solid (460 mg, 51% yield). LC-MS: calculated exact mass: 443.1 experimental value: [M+H] + (ESI) = 443.6; 1 H NMR (400 MHz, chloroform-d) δ ppm: 8.22 (s, 1H), 7.79 (s, 1H), 5.70 (br.s., 2H), 4.72 - 4.81 (m, 1H), 4.51 (m, 1H), 4.15 - 4.27 (m, 4H), 3.11 (d, J = 10.4 Hz, 2H), 2.40 - 2.52 (m, 4H), 2.10 - 2.25 (m, 2H), 1.86 - 1.96 (m, 2H), 1.55 - 1.81 (m, 2H).

[0328] Example 1

[0329] 7-((trans)-4-(4-methylpiperazin-1-yl)cyclohexyl)-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0330] and

[0331] Example 2

[0332] 7-((cis)-4-(4-Methylpiperazin-1-yl)cyclohexyl)-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0333]

[0334] A solution of 4-(4-amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexanone (500 mg, 1.25 mmol), 1-methylpiperazine (0.4 mL, 3.76 mmol), acetic acid (0.2 mL, 1.88 mmol) and molecular sieve (1.0 g) in 1,2-dichloroethane (20 mL) was stirred at room temperature for 5 h. Subsequently, sodium triacetoxyborohydride (398 mg, 1.88 mmol) was added and stirring was continued at room temperature for 16 h. Water (100 mL) and dichloromethane (100 mL) were added to the reaction mixture, saturated aqueous sodium bicarbonate solution (100 mL) was further added thereto and the mixture was partitioned. The organic layer was washed with brine (60 mL) and dried over sodium sulfate, and concentrated and purified by flash chromatography (DCM:MeOH = 10:1) to obtain 7-((trans)-4-(4-methylpiperazin-1-yl)cyclohexyl)-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (100 mg, 16.5% yield) as a yellow solid. LCMS: calculated exact mass = 482.3; experimental value [M+H] + (ESI) = 483.3; 1 1H NMR (400 MHz, chloroform-d) δ ppm: 8.34 (s, 1H), 7.36 - 7.48 (m, 4H), 7.15 - 7.20 (m, 1H), 7.07 - 7.13 (m, 4H), 7.02 (s, 1H), 5.18 (br.s., 2H), 4.70 (tt, J = 12.1, 3.8 Hz, 1H), 2.76 (br.s., 4H), 2.63 (br.s., 2H), 2.49 - 2.60 (m, 2H), 2.36 - 2.45 (m, 3H), 2.21 - 2.33 (m, 2H), 2.13 (d, J = 12.6 Hz, 2H), 1.76 - 1.92 (m, 2H), 1.57 - 1.69 (m, 2H).

[0335] Data of 7-((cis)-4-(4-methylpiperazin-1-yl)cyclohexyl)-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (80 mg, 13.2%) LCMS: calculated exact mass = 482.3; experimental value [M+H] + (ESI) = 483.3; 11H NMR (400 MHz, chloroform-d) δ ppm: 8.29 - 8.40 (m, 1H), 7.45 - 7.54 (m, 2H), 7.36 - 7.45 (m, 2H), 7.17 (t, J = 7.4 Hz, 1H), 7.07 - 7.15 (m, 5H), 5.13 (s, 2H), 4.80 - 4.90 (m, 1H), 2.58 - 2.73 (m, 7H), 2.30 - 2.41 (m, 4H), 2.16 - 2.28 (m, 5H), 1.86 (dd, J = 8.6, 3.8 Hz, 2H), 1.68 (t, J = 13.7 Hz, 2H).

[0336] Using a similar procedure, the following compounds can be obtained:

[0337]

[0338]

[0339]

[0340]

[0341]

[0342]

[0343]

[0344]

[0345]

[0346]

[0347]

[0348]

[0349]

[0350]

[0351]

[0352]

[0353]

[0354]

[0355]

[0356]

[0357]

[0358]

[0359]

[0360]

[0361]

[0362]

[0363]

[0364]

[0365]

[0366] Example 86

[0367] 6-Bromo-7-((trans)-4-(4-methylpiperazin-1-yl)cyclohexyl)-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0368]

[0369] To a solution of 7-((trans)-4-(4-methylpiperazin-1-yl)cyclohexyl)-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (500 mg, 10.4 mmol) in dichloromethane (15 mL) was added NBS (368.8 mg, 2.07 mmol) portionwise at room temperature. Subsequently, the mixture was stirred for an additional 16 h. The mixture was concentrated and purified by flash chromatography (DCM:MeOH = 15:1) to afford a yellow solid (280 mg, 48.3% yield). LCMS: calculated exact mass = 560.2; found [M+H]+ (ESI) = 561.8; 11H NMR (400 MHz, chloroform-d) δ ppm: 8.23 (br.s., 1H), 7.33 - 7.45 (m, 4H), 7.16 - 7.23 (m, 1H), 7.11 (dd, J = 7.8, 5.3 Hz, 4H), 5.48 (br.s., 2H), 4.61 (br.s., 1H), 3.12 - 3.35 (m, 6H), 2.81 - 3.08 (m, 2H), 2.75 - 2.80 (m, 2H), 2.66 (br.s., 3H), 2.24 (br.s., 2H), 2.03 (d, J = 11.3 Hz, 2H), 1.69 - 1.77 (m, 2H).

[0370] Example 87

[0371] 7 - ((trans)-4-(4-Methylpiperazin-1-yl)cyclohexyl)-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-ol

[0372] and

[0373] Example 88

[0374] 7 - ((cis)-4-(4-Methylpiperazin-1-yl)cyclohexyl)-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-ol

[0375]

[0376] Under a N 2 atmosphere, a solution of 4-chloro-5-(4-phenoxyphenyl)-7-(1,4-dioxaspiro[4.5]dec-8-yl)-7H-pyrrolo[2,3-d]pyrimidine (885 mg, 1.92 mmol) in MeOH (20 mL) at room temperature, KOH (215 mg, 3.83 mmol) was added and the mixture was stirred at 80 °C for 4 h. TLC (PE:EA = 4:1) showed complete depletion of the starting material. The reaction mixture was cooled to room temperature, poured into ice water (50 mL) and extracted with EA (50 mL × 2). The organic layer was washed with brine, dried over anhydrous Na 2 SO 4 and concentrated in vacuo to give the crude product as a yellow solid (623 mg, 71% yield), which was used in the next step without further purification.

[0377] LCMS: Calculated exact mass = 457.20, experimental value [M+H] + (ESI) = 458.2

[0378] 4-(4-Hydroxy-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexanone

[0379] A solution of 4-methoxy-5-(4-phenoxyphenyl)-7-(1,4-dioxaspiro[4.5]dec-8-yl)-7H-pyrrolo[2,3-d]pyrimidine (623 mg, 1.36 mmol) in THF (3 mL), acetone (15 mL) and 6 M HCl (11 mL) was stirred at 80 °C for 4 h. TLC (EA:PE = 1:2) showed complete depletion of the starting material. The mixture was cooled in an ice bath. 1 M NaOH (80 mL, aqueous solution) was added until pH 8. The light brown solid product (465 mg, 85.5% yield) was collected by filtration and used in the next step without further purification. LCMS: calculated exact mass = 399.16, experimental value [M+H] + (ESI) = 400.1.

[0380] 7-((cis)-4-(4-Methylpiperazin-1-yl)cyclohexyl)-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-ol

[0381] and

[0382] 7-((trans)-4-(4-Methylpiperazin-1-yl)cyclohexyl)-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-ol

[0383] At room temperature, to a solution of compound 4-(4-hydroxy-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexanone (100 mg, 0.25 mmol), 1-methylpiperazine (200 mg, 2.00 mmol) and ZnCl 2 (102 mg, 0.75 mmol in MeOH (20 mL) was added NaBH 3 CN (47 mg, 0.75 mmol) and AcOH (0.1 mL) was added under N 2 atmosphere. Subsequently, the reaction mixture was stirred at 40 °C for 16 h. After concentration, the residue was extracted with DCM (150 mL×2) and water. The combined organic layers were washed with an aqueous solution of NH 4 Cl (200 mL), dried over anhydrous Na 2 SO 4Dry and concentrate in vacuo. Purify the residue by Prep-TLC and subsequently by Prep-HPLC to afford 7-(cis)-4-(4-methylpiperazin-1-yl)cyclohexyl)-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-ol as a white solid (10 mg, 8.3% yield). LCMS: calculated exact mass = 483.26, found [M+H] + (ESI)=484.2; 1 HNMR(400MHz,DMSO)δ11.99(s,1H),10.40(s,1H),7.94(dd,J = 17.1,5.5Hz,3H),7.41(dd,J = 14.4,5.9Hz,3H),7.14(t,J = 7.4Hz,1H),7.02(dd,J = 7.9,6.6Hz,4H),4.64(s,1H),3.05(s,2H),2.65(s,3H),2.27(d,J = 17.6Hz,2H),2.03(dd,J = 23.5,9.1Hz,4H),1.71(s,2H),1.59(s,2H),1.30 - 1.19(m,5H). Data for 7-(trans)-4-(4-methylpiperazin-1-yl)cyclohexyl)-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-ol: 1 H NMR(400MHz,DMSO)δ8.92(s,1H),7.95(s,1H),7.42 - 7.32(m,7H),7.18 - 7.07(m,2H),7.05 - 6.93(m,7H),6.85(d,J = 8.1Hz,4H),4.83(s,1H),3.54 - 3.45(m,2H),2.79(s,3H),2.12(d,J = 23.4Hz,7H),1.69(s,2H),1.23(s,5H).

[0384] Example 89

[0385] 1-(4-((trans)-4-(4-amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexyl)piperazin-1-yl)-2-methylpropan-2-ol

[0386]

[0387] 5-(4-phenoxyphenyl)-7-((trans)-4-(piperazin-1-yl)cyclohexyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0388] To a solution of tert-butyl 4-((trans)-4-(4-amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexyl)piperazine-1-carboxylate (170 mg, 0.3 mmol) in DCM (4 mL) was added dropwise TFA (2 mL). The reaction mixture was stirred at room temperature for 1 h. The reaction mixture was concentrated to afford the product (50 mg, 35% yield). LCMS: calculated exact mass = 568.3; experimental value [M+H] + (ESI) = 569.2.

[0389] 1-(4-((trans)-4-(4-amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexyl)piperazin-1-yl)-2-methylpropan-2-ol

[0390] A reaction mixture of 5-(4-phenoxyphenyl)-7-((trans)-4-(piperazin-1-yl)cyclohexyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (100 mg, 0.21 mmol), 2,2-dimethyloxirane (30 mg, 0.43 mmol) and Cs 2 CO 3 (208 mg, 0.64 mmol) in DMF (8 mL) was stirred at room temperature for 2 h. NaBH 3 CN (47 mg, 0.75 mol) was added. It was stirred at 100 °C overnight. After cooling to room temperature, DCM (20 mL) was added and the mixture was washed with water (20 mL×2). The organic layer was collected and concentrated. The residue was purified by Prep-TLC (DCM:MeOH = 10:1) to afford the desired product (6 mg, 5% yield). LCMS: calculated exact mass = 540.2; experimental value [M+H] + (ESI) = 540.3; 1 H NMR (400 MHz, DMSO-d 6 ) δ ppm: 10.46 (br.s., 1H) 8.14 (s, 1H) 7.38 - 7.50 (m, 6H) 7.16 (t, J = 7.38 Hz, 2H) 7.10 (dd, J = 8.07, 6.07 Hz, 6H) 6.15 (s, 1H) 4.58 - 4.61 (m, 1H) 4.21 - 4.26 (m, 1H) 3.01 - 3.09 (m, 4H) 2.57 - 2.71 (m, 2H) 2.20 - 2.36 (m, 4H) 1.88 - 2.08 (m, 4H) 1.75 - 1.82 (m, 2H) 1.10 (s, 6H).

[0391] Example 90

[0392] 1-((trans)-4-(4-Amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexyl)piperazin-2-one

[0393] and

[0394] Example 91

[0395] 1-((cis)-4-(4-Amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexyl)piperazin-2-one

[0396] tert-Butyl (tert-butoxycarbonyl)(7-(4-oxocyclohexyl)-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)carbamate

[0397] To a solution of 4-(4-amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexan-1-one (2 g, 5.01 mmol) in THF (10 mL) was added Et 3 N (1.5 g, 15.06 mmol) and (Boc) 2 O (2.2 g, 10 mmol). The mixture was stirred overnight at room temperature. DMAP (122 mg, 1 mmol), Et 3 N (1.5 g, 15.06 mmol) and (Boc) 2 O (2.2 g, 10 mmol) were added and the mixture was refluxed for 3 h. The reaction was monitored by TLC and LCMS until the starting material was completely consumed. It was then concentrated and washed with saturated NaHCO 3 and extracted with DCM. The organic layer was concentrated to give the crude product. The crude product was purified by flash column chromatography (0 - 0.5% MeOH / DCM) to give the product as a yellow solid (1.5 g, 50% yield). LCMS: calculated exact mass = 598.69; experimental value [M+H] + (ESI) = 599.11.

[0398] tert-Butyl (tert-butoxycarbonyl)(7-(4-((2-((tert-butoxycarbonyl)amino)ethyl)amino)cyclohexyl)-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)carbamate

[0399] A solution of tert-butyl (tert-butoxycarbonyl)(7-(4-oxocyclohexyl)-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)carbamate (1250 mg, 2.088 mmol) and tert-butyl (2-aminoethyl)carbamate (2007 mg, 12.52 mmol) in DCE (15 mL) was stirred at 50 °C under N 2 atmosphere for 2 h. After cooling it to room temperature, NaBH(OAc) 3 (2655 mg, 12.527 mmol) was added portionwise, and the mixture was stirred at room temperature under N 2 atmosphere overnight. The reaction was monitored by TLC and LCMS until the starting materials were completely exhausted. Then it was filtered and concentrated, and the crude product was purified by flash column chromatography (2-3% MeOH / DCM) to obtain the product as a white solid (1 g, 64.5% yield). LCMS: calculated exact mass = 742.41; experimental value [M+H] + (ESI) = 742.92.

[0400] tert-Butyl (tert-butoxycarbonyl)(7-(4-(N-(2-((tert-butoxycarbonyl)amino)ethyl)-2-chloroacetamido)cyclohexyl)-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)carbamate

[0401] At 0 °C, Et 3 N (430 mg, 2.05 mmol) and 2-chloroacetyl chloride (98.82 mg, 0.875 mmol) in DCM (0.5 mL) were added dropwise to a solution of tert-butyl (tert-butoxycarbonyl)(7-(4-((2-((tert-butoxycarbonyl)amino)ethyl)amino)cyclohexyl)-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)carbamate (500 mg, 0.67 mmol) in DCM (5 ml). The mixture was warmed to room temperature for 2 h. The reaction was monitored by LCMS until the starting materials were completely exhausted. The reaction mixture was quenched with ice / water and extracted with DCM (20 mL×2). Then it was concentrated to obtain the crude product. The crude product was purified by flash column chromatography (1-1.5% MeOH / DCM) to obtain the product as a colorless solid (175 mg, 31.7% yield). LCMS: calculated exact mass = 818.38; experimental value [M+H] + (ESI) = 818.9.

[0402] tert-Butyl (tert-butyl) 4-(4-(4-((tert-butoxycarbonyl)amino)-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexyl)-3-oxopiperazine-1-carboxylate

[0403] To a solution of tert-butyl (7-(4-(N-(2-((tert-butoxycarbonyl)amino)ethyl)-2-chloroacetamido)cyclohexyl)-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)carbamate (175 mg, 0.214 mmol) in DMF (2 mL) at 0 °C was added (33 mg, 60% in oil, 0.854 mmol), and then the temperature was raised to room temperature for 2 h. The reaction was monitored by TLC and LCMS until the starting material was completely consumed. The reaction mixture was quenched with ice / water and extracted with DCM (50 mL × 5), and the organic layer was concentrated to give the product as a yellow solid (150 mg, 89.5% yield). LCMS: calculated exact mass = 782.4; experimental value [M+H] + (ESI) = 682.62.

[0404] 1-((trans)-4-(4-Amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexyl)piperazin-2-one and 1-((cis)-4-(4-amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexyl)piperazin-2-one

[0405] To a solution of tert-butyl (tert-butyl) 4-(4-(4-((tert-butoxycarbonyl)amino)-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexyl)-3-oxopiperazine-1-carboxylate (150 mg, 0.19 mmol) in DCM (2 ml) was added TFA (2 mL). The mixture was stirred at room temperature for 3 h. The reaction was monitored by TLC and LCMS until the starting material was completely consumed. The mixture was concentrated and 20 mL of 7.0 N NH 3 / MeOH was added, and the mixture was stirred at room temperature for 15 min. The mixture was concentrated and prep-TLC was performed to give 1-((trans)-4-(4-amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexyl)piperazin-2-one as a yellow solid (45 mg, 48.6% yield). LCMS: calculated exact mass = 482.24; experimental value [M+H] + (ESI) = 482.8; 1 H NMR (DMSO-d 6) δ ppm: 8.14 (s, 1H), 7.38 - 7.52 (m, 5H), 7.16 (t, J = 7.3 Hz, 1H), 7.06 - 7.12 (m, 4H), 6.03 - 6.24 (m, 1H), 4.62 (t, J = 11.7 Hz, 1H), 4.41 - 4.51 (m, 1H), 3.25 (s, 2H), 3.21 (t, J = 5.0 Hz, 2H), 2.88 (t, J = 5.2 Hz, 2H), 2.05 (d, J = 9.5 Hz, 2H), 1.99 (br.s., 2H), 1.72 - 1.86 (m, 2H), 1.66 (d, J = 10.7 Hz, 2H).

[0406] Data of 1 - ((cis) - 4 - (4 - amino - 5 - (4 - phenoxyphenyl) - 7H - pyrrolo[2,3 - d]pyrimidin - 7 - yl)cyclohexyl)piperazin - 2 - one (300 mg, 48.6% yield). LCMS: Calculated exact mass = 482.24; experimental value [M + H] + (ESI) = 482.79; 1 H NMR (DMSO - d 6 ) δ ppm: 8.14 (s, 1H), 7.48 - 7.56 (m, 3H), 7.38 - 7.46 (m, 2H), 7.06 - 7.20 (m, 5H), 6.12 (br.s., 1H), 4.80 (d, J = 3.7 Hz, 1H), 4.30 - 4.40 (m, 1H), 3.23 - 3.28 (m, 4H), 2.89 (t, J = 5.2 Hz, 2H), 2.37 (d, J = 11.0 Hz, 2H), 1.95 - 2.04 (m, 2H), 1.80 - 1.91 (m, 2H), 1.57 (d, J = 9.2 Hz, 2H).

[0407] Example 92

[0408] 7 - (3 - amino - 1,5 - dioxaspiro[5.5]undecan - 9 - yl) - 5 - (4 - phenoxyphenyl) - 7H - pyrrolo[2,3 - d]pyrimidin - 4 - amine

[0409]

[0410] (9 - (4 - amino - 5 - (4 - phenoxyphenyl) - 7H - pyrrolo[2,3 - d]pyrimidin - 7 - yl) - 1,5 - dioxaspiro[5.5]undecan - 3 - yl)carbamic acid tert - butyl ester

[0411] To a solution of 4-(4-amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexanone (400 mg, 1 mmol) in 10 mL of toluene was added tert-butyl (1,3-dihydroxypropan-2-yl)carbamate (191 mg, 1 mmol), TsOH (12 mg, 0.06 mmol), and molecular sieves. The reaction mixture was heated to 150 °C and reacted under microwave irradiation for 2 hours. The solvent was removed under reduced pressure. The crude product was purified by flash column chromatography (EA 100%) to give the product as a white solid (160 mg, 28% yield). LC-MS: calculated exact mass = 571.3, experimental value [M+H] + (ESI) = 572.12.

[0412] 7-(3-Amino-1,5-dioxaspiro[5.5]undecan-9-yl)-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0413] To a solution of tert-butyl (9-(4-amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-1,5-dioxaspiro[5.5]undecan-3-yl)carbamate (260 mg, 0.45 mmol) in 20 mL of DCM at -78 °C was added 2,6-dimethylpyridine (72 mg, 0.675 mmol) and TMSOTf (130 mg, 0.585 mmol). The dry ice / acetone bath was replaced with an ice water bath, and the reaction mixture was stirred for 2 hours, followed by the addition of NaHCO 3 (20 mL aqueous solution) and extracted with EA (20 mL × 3). The organic layers were combined and dried over Na 2 SO 4 After concentration, the crude product was purified by Prep-TLC (DCM:CH 3 OH = 10:1) to give the product as a white solid (19 mg, 8.6% yield). LC-MS: calculated exact mass = 471.2, experimental value [M+H] + (ESI) = 472.11; 1 H NMR (DMSO-d 6)δ ppm: 8.14 (s, 1H), 7.48 (d, J = 8.7 Hz, 2H), 7.39 - 7.44 (m, 2H), 7.36 (s, 1H), 7.16 (t, J = 7.3 Hz, 1H), 7.06 - 7.11 (m, 4H), 4.60 - 4.73 (m, 1H), 3.97 (dd, J = 11.7, 3.4 Hz, 2H), 3.59 - 3.68 (m, 2H), 2.96 (br.s., 1H), 2.45 (d, J = 13.5 Hz, 1H), 2.37 (d, J = 13.5 Hz, 1H), 1.92 - 2.03 (m, 3H), 1.86 (d, J = 9.8 Hz, 2H), 1.54 (ddt, J = 17.3, 13.6, 3.7 Hz, 2H).

[0414] Example 93

[0415] 8-(4-Amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-3-(pyridin-4-yl)-1,3-diazaspiro[4.5]decane-2,4-dione

[0416]

[0417] 8-(4-Amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-1,3-diazaspiro[4.5]decane-2,4-dione

[0418] To a solution of 4-(4-amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexan-1-one (5 g, 12.6 mmol) in EtOH (50 mL) and water (50 mL) was added NaCN (926 mg, 18.9 mmol) and (NH 4 ) 2 CO 3 (3.63 g, 37.8 mmol). The mixture was stirred at 80 °C overnight. Then it was cooled to room temperature and concentrated to remove EtOH, filtered and washed with water (10 mL), and the solid was dried to give the product (4.5 g, 76% yield). LCMS: Calculated exact mass = 468.2; Experimental value [M+H] + (ESI) = 469.2.

[0419] 8-(4-Amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-3-(pyridin-4-yl)-1,3-diazaspiro[4.5]decane-2,4-dione

[0420] To a solution of 8-(4-amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-1,3-diazaspiro[4.5]decane-2,4-dione (230 mg, 0.5 mmol) and pyridin-4-ylboronic acid (95 mg, 0.75 mmol) in 15 mL of DCE was added Cu(OAc) 2 (97 mg, 0.5 mmol) and Et 3 N (0.15 mL, 1 mmol). The reaction mixture was stirred overnight at room temperature under an O 2 atmosphere. The reaction mixture was filtered through celite and the solid was washed with 10 mL of MeOH. The filtrate was concentrated and purified by Prep-TLC (DCM:CH 3 OH = 10:1) to give 70 mg of the crude product, which was further purified by Prep-HPLC to give the product as a white solid (11 mg, 4% yield). LC-MS: calculated exact mass = 545.2, experimental value [M+H] + (ESI) = 545.8; 1 H NMR (methanol-d 4 ) δ ppm: 8.78 (br.s., 2H), 8.48 (d, J = 5.8 Hz, 2H), 8.27 (s, 1H), 7.53 (s, 1H), 7.42 (d, J = 8.5 Hz, 2H), 7.28 - 7.35 (m, 2H), 6.96 - 7.12 (m, 6H), 1.97 - 2.25 (m, 9H), 1.93 (d, J = 6.1 Hz, 1H).

[0421] Example 94

[0422] 2-(4-(4-Amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexylidene)acetamide

[0423] and

[0424] Example 95

[0425] Methyl 2-(1-amino-4-(4-amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexyl)acetate

[0426] and

[0427] Example 96

[0428] 2-(1-Amino-4-(4-amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexyl)acetamide

[0429]

[0430] Ethyl 2-(4-(4-amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexylidene)acetate

[0431] 4-(4-Amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexanone (1.5 g, 3.76 mmol) and ethyl 2-(triphenylphosphoranylidene)acetate (1.3 g, 3.76 mmol) were reacted in toluene (100 mL). The reaction mixture was stirred overnight at 110 °C. The reaction mixture was cooled and concentrated. The residue was purified by flash column chromatography (DCM:EtOAc = 1:1) to give the product (900 mg, 51% yield).

[0432] LCMS: Calculated exact mass = 468.2; experimental value [M+H] + (ESI) = 468.6.

[0433] 2-(4-(4-Amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexylidene)acetamide

[0434] and

[0435] Methyl 2-(1-amino-4-(4-amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexyl)acetate

[0436] and

[0437] 2-(1-Amino-4-(4-amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexyl)acetamide

[0438] A solution of ethyl 2-(4-(4-amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexylidene)acetate (350 mg, 0.75 mmol) in NH 3 -MeOH (14 mL) was reacted in a microwave reactor at 100 °C for 48 h. After cooling to room temperature, the reaction mixture was concentrated. The residue was purified by column chromatography (DCM:MeOH = 10:1) to give three products: 2-(4-(4-amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexylidene)acetamide (2 mg, 0.6% yield). LCMS: Calculated exact mass = 439.2; experimental value [M+H] + (ESI) = 439.6; 1HNMR(400MHz,DMSO-d 6 )δppm:8.13(s,1H)7.47(d,J=8.55Hz,2H)7.38-7.45(m,3H)7.32(br.s.,1H)7.16(t,J=7.48Hz,1H)7.05-7.13(m,4H)6.85(br.s.,1H)5.50-5.60(m,1H)4.83(br.s.,1H)2.80(s,2H)2.45(br.s.,1H)2.33(br.s.,1H)2.24(br.s.,1H)2.15(d,J=14.65Hz,2H)1.89-2.06(m,2H)1.81(br.s.,1H).

[0439] 2-(1-Amino-4-(4-amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexyl)acetate (2 mg, 0.6% yield). LCMS: calculated exact mass = 471.2; experimental value [M+H] + (ESI)=471.6; 1 H NMR(400MHz,DMSO-d 6 )δppm:8.13(s,1H)7.67(s,1H)7.48(d,J=8.54Hz,2H)7.42(t,J=7.93Hz,2H)7.16(t,J=7.48Hz,1H)7.08(d,J=7.93Hz,2H)7.11(d,J=8.54Hz,2H)6.14(br.s.,1H)4.61(br.s.,1H)3.68(s,3H)3.01(s,2H)1.95-2.14(m,4H)1.72-1.92(m,4H).

[0440] 2-(1-Amino-4-(4-amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexyl)acetamide (2 mg, 6% yield). LCMS: calculated exact mass = 456.2; experimental value [M+H] + (ESI)=456.8; 1 H NMR(400MHz,DMSO-d 6)δ ppm: 8.13 (s, 1H), 7.95 (s, 2H), 7.99 (s, 1H), 7.56 - 7.63 (m, 1H), 7.48 (d, J = 8.24 Hz, 3H), 7.42 (t, J = 7.78 Hz, 2H), 7.17 (d, J = 7.32 Hz, 1H), 7.03 - 7.15 (m, 4H), 6.16 (br.s., 1H), 4.62 (br.s., 1H), 2.89 (s, 1H), 2.70 - 2.80 (m, 2H), 1.75 - 2.05 (m, 7H).

[0441] Example 97

[0442] 2-(4-(4-Amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexylidene)-N-methylacetamide

[0443] and

[0444] Example 98

[0445] 2-(4-(4-Amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-1-(methylamino)cyclohexyl)-N-methylacetamide

[0446]

[0447] A solution of ethyl 2-(4-(4-amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexylidene)acetate (40 mg, 0.085 mmol) and methylamine (40% in water, 0.5 mL) in MeOH (2 mL) was reacted in a microwave reactor at 100 °C for 2 hours. After cooling to room temperature, the reaction mixture was concentrated. The residue was purified by column chromatography (DCM:MeOH = 10:1) to give two products: 2-(4-(4-amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexylidene)-N-methylacetamide (3 mg, 7.8% yield). LCMS: calculated exact mass = 453.2; experimental value [M+H] + (ESI) = 453.8; 1 H NMR (400 MHz, DMSO-d 6)δ ppm: 8.13 (s, 1H) 7.79 (d, J = 4.27 Hz, 1H) 7.44 - 7.50 (m, 3H) 7.37 - 7.44 (m, 3H) 7.16 (t, J = 7.17 Hz, 2H) 7.09 (dd, J = 8.09, 4.43 Hz, 5H) 7.01 (br.s., 1H) 5.55 (br.s., 1H) 4.81 (br.s., 1H) 2.81 (s, 2H) 2.56 - 2.61 (m, 3H) 2.44 (br.s., 2H) 2.22 (br.s., 1H) 2.04 - 2.18 (m, 2H) 1.89 - 2.04 (m, 3H); and 2-(4-(4-amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-1-(methylamino)cyclohexyl)-N-methylacetamide (3 mg, 7.2% yield). LCMS: calculated exact mass = 484.2; experimental value [M + H] + (ESI) = 484.8; 1 H NMR (400 MHz, DMSO-d 6 )δ ppm: 8.50 (s, 1H) 8.14 (s, 1H) 7.55 (s, 1H) 7.49 (d, J = 8.54 Hz, 2H) 7.42 (t, J = 7.63 Hz, 2H) 7.03 - 7.21 (m, 5H) 4.63 (br.s., 1H) 2.85 (br.s., 2H) 2.68 (d, J = 4.27 Hz, 3H) 2.60 (s, 3H) 1.90 (s, 2H) 1.94 (s, 3H) 1.81 (br.s., 2H) 1.75 (s, 1H).

[0448] Example 99

[0449] Ethyl 6-(4-amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)spiro[2.5]octane-1-carboxylate

[0450]

[0451] Ethyl 6-(4-amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)spiro[2.5]octane-1-carboxylate

[0452] When adding ethyl 2-(4-(4-amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexylidene)acetate (100 mg, 0.21 mmol), a solution of trimethylsulfoxonium iodide (92 mg, 0.42 mmol) and t-BuOK (47 mg, 0.42 mmol) in DMSO (2 mL) was stirred at room temperature for 3 hours. The reaction mixture was stirred overnight. Subsequently, it was diluted with EtOAc, washed with water and brine, dried and concentrated. The residue was purified by Prep-TLC (DCM:MeOH = 20:1) to give the product (10 mg, 4.9% yield). LCMS: calculated exact mass = 482.2; experimental value [M+H] + (ESI) = 483.1; 1 H NMR (400 MHz, CDCl 3 -d 6 ) δ ppm: 8.30 (s, 1H) 7.40 - 7.46 (m, 4H) 2.15 - 2.19 (m, 3H) 1.95 - 1.99 (m, 3H) 1.57 - 1.60 (m, 2H) 1.33 (s, 3H) 1.10 - 1.31 (m, 3H).

[0453] Example 100

[0454] 4-(4-Amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-1-((4-methylpiperazin-1-yl)methyl)cyclohexanol

[0455]

[0456] 5-(4-Phenoxyphenyl)-7-(1-oxaspiro[2.5]octan-6-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0457] When adding ethyl 2-(4-(4-amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexylidene)acetate (200 mg, 0.50 mmol), a solution of trimethylsulfoxonium iodide (220 mg, 1.00 mmol) and t-BuOK (110 mg, 1.00 mmol) in DMSO (10 mL) was stirred at room temperature for 0.5 hour. The reaction mixture was stirred overnight. The reaction was quenched with ice water, extracted with DCM, dried and concentrated to give the crude product (250 mg), which was used without further purification. LCMS: calculated exact mass = 412.1; experimental value [M+H]+ (ESI) = 412.8.

[0458] 4-(4-Amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-1-((4-methylpiperazin-1-yl)methyl)cyclohexanol

[0459] A solution of 5-(4-phenoxyphenyl)-7-(1-oxaspiro[2.5]octan-6-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (50 mg, 0.12 mmol) and 1-methylpiperazine (50 mg, 0.50 mmol) in MeOH (10 mL) was heated at 70 °C for 5 h. After cooling to room temperature, the reaction mixture was concentrated. The residue was purified by flash column chromatography (DCM:MeOH = 10:1) to give the product (20 mg, 39% yield over two steps). LCMS: calculated exact mass = 512.2; experimental value [M+H] + (ESI) = 512.8; 1 H NMR (400 MHz, CDCl 3 -d 6 ) δ ppm: 8.13 (s, 1H) 7.48 (d, J = 8.55 Hz, 2H) 7.39 - 7.45 (m, 2H) 7.37 (s, 1H) 7.13 - 7.19 (m, 1H) 7.09 (d, J = 8.55 Hz, 4H) 6.12 (br.s., 1H) 4.53 (t, J = 12.21 Hz, 1H) 4.11 (s, 1H) 2.54 (br.s., 3H) 2.35 (br.s., 4H) 2.26 (s, 3H) 2.08 - 2.20 (m, 5H) 1.63 - 1.79 (m, 4H) 1.49 - 1.63 (m, 2H).

[0460] Example 101

[0461] 1-((1H-Imidazol-1-yl)methyl)-4-(4-amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexanol

[0462]

[0463] 1-((1H-Imidazol-1-yl)methyl)-4-(4-amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexanol

[0464] A solution of 5-(4-phenoxyphenyl)-7-(1-oxaspiro[2.5]octan-6-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (40 mg, 0.10 mmol), 1H-imidazole (27 mg, 0.40 mmol) and K 2 CO 3A solution of 2 SO 4 (50 mg, 0.40 mmol) in DMF (5 mL) was heated at 100 °C for 18 h. After cooling to room temperature, the reaction mixture was concentrated. The residue was dissolved in DCM, washed with water, dried over Na + (ESI)=480.8; 1 H NMR (400 MHz, CDCl 3 -d 6 ) δ ppm: 8.05 (s, 1H) 7.60 (s, 1H) 7.33 - 7.43 (m, 4H) 7.32 (s, 1H) 7.08 - 7.16 (m, 2H) 7.01 (d, J = 7.63 Hz, 2H) 7.04 (d, J = 8.54 Hz, 3H) 6.89 (s, 1H) 4.5 (s, 3H) 3.94 (s, 2H) 1.87 - 2.07 (m, 3H) 1.73 (d, J = 11.29 Hz, 2H) 1.60 (t, J = 12.05 Hz, 2H) 1.50 (d, J = 12.51 Hz, 3H).

[0465] Example 102

[0466] 1 - ((1H - Tetrazol - 1 - yl)methyl)-4-(4 - amino - 5-(4 - phenoxyphenyl)-7H - pyrrolo[2,3 - d]pyrimidin - 7 - yl)cyclohexanol

[0467]

[0468] 1 - ((1H - Tetrazol - 1 - yl)methyl)-4-(4 - amino - 5-(4 - phenoxyphenyl)-7H - pyrrolo[2,3 - d]pyrimidin - 7 - yl)cyclohexanol

[0469] A solution of 5-(4 - phenoxyphenyl)-7-(1 - oxaspiro[2.5]octan - 6 - yl)-7H - pyrrolo[2,3 - d]pyrimidin - 4 - amine (50 mg, 0.12 mmol), 1H - tetrazole (34 mg, 0.48 mmol) and K 2 CO 3 (66 mg, 0.48 mmol) in DMF (5 mL) was heated at 100 °C for 4 h. After cooling to room temperature, the reaction mixture was concentrated. The residue was dissolved in DCM, washed with water, dried over Na 2 SO 4Dried, concentrated and purified by Prep-TLC (DCM:MeOH = 10:1) to give the product (3 mg, 5% yield). LCMS: calculated exact mass = 482.2; experimental value [M+H] + (ESI) = 482.8; 1 H NMR (400 MHz, methanol-d 4 ) δ ppm: 9.07 (s, 1H) 8.05 (s, 1H) 7.37 (d, J = 8.85 Hz, 2H) 7.28 (t, J = 7.93 Hz, 2H) 7.17 (s, 1H) 7.04 (t, J = 7.48 Hz, 1H) 6.96 (d, J = 7.93 Hz, 2H) 6.99 (d, J = 8.85 Hz, 2H) 4.58 (t, J = 12.66 Hz, 1H) 4.45 (s, 2H) 1.99 - 2.14 (m, 3H) 1.92 (s, 1H) 1.84 (s, 2H) 1.67 - 1.83 (m, 4H) 1.58 (d, J = 13.12 Hz, 3H).

[0470] Example 103

[0471] 3-(2-Fluoro-4-phenoxyphenyl)-1-((trans)-4-(4-methylpiperazin-1-yl)cyclohexyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine

[0472]

[0473] 3-(2-Fluoro-4-phenoxyphenyl)-1-((trans)-4-(4-methylpiperazin-1-yl)cyclohexyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine

[0474] To a 500 mL round-bottom flask was added 3-iodo-1-((trans)-4-(4-methylpiperazin-1-yl)cyclohexyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (25 g, 56.6 mmol), 2-(2-fluoro-4-phenoxyphenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (21.35 g, 67.9 mmol), Pd(dppf)Cl 2 (4.14 g, 5.6 mmol), Na 2 CO 3(24 g, 226.5 mmol) dioxane - water (9 - 1, 300 mL). The reactants were stirred overnight at 85 °C. It was cooled to room temperature, filtered and concentrated, and the crude product was purified by flash column chromatography (MeOH / DCM, 0 to 15% gradient) to obtain the product (15 g) as a white solid. The product was further purified by Prep - HPLC (acetonitrile - water gradient, containing 0.1% TFA). After lyophilization, the product was obtained as a white powder (10.3 g, 41.2% yield). LC - MS: Calculated exact mass: 501.27; Experimental value [M + H] + (ESI) = 502.32; 1 1H NMR (400 MHz, DMSO - d6) δ ppm: 8.32 (s, 1H), 7.54 (t, J = 8.6 Hz, 1H), 7.45 - 7.51 (m, 2H), 7.22 - 7.29 (m, 1H), 7.15 - 7.22 (m, 2H), 7.03 (dd, J = 11.3, 2.4 Hz, 1H), 6.96 (dd, J = 8.5, 2.4 Hz, 1H), 4.75 (dd, J = 9.8, 4.8 Hz, 1H), 3.51 (br.s., 4H), 3.18 (br.s., 5H), 2.82 (s, 3H), 1.98 - 2.24 (m, 6H), 1.69 (d, J = 6.6 Hz, 2H).

[0475] Using a similar procedure, the following compounds can be prepared:

[0476]

[0477]

[0478]

[0479]

[0480]

[0481]

[0482]

[0483]

[0484]

[0485]

[0486]

[0487] 4-Chloro-5-(3-methoxy-4-phenoxyphenyl)-7-(1,4-dioxaspiro[4.5]dec-8-yl)-7H-pyrrolo[2,3-d]pyrimidine

[0488] A mixture of 4-chloro-5-iodo-7-(1,4-dioxaspiro[4.5]dec-8-yl)-7H-pyrrolo[2,3-d]pyrimidine (800 mg, 1.9 mmol), 2-(3-methoxy-4-phenoxyphenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (1.3 g, 3.98 mmol), Pd(dppf)Cl 2 (140 mg, 0.19 mmol), and Na 2 CO 3 (600 mg, 5.7 mmol) in dioxane-H 2 O (50 mL - 5 mL) was heated at 80 °C for 3 h under an inert atmosphere. After cooling to room temperature, the reaction mixture was concentrated and purified by flash column chromatography (EA / PE, 0 to 33% gradient) to give the product as a brown solid (350 mg, 37% yield).

[0489] 5-(3-Methoxy-4-phenoxyphenyl)-7-(1,4-dioxaspiro[4.5]dec-8-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0490] A mixture of 4-chloro-5-(3-methoxy-4-phenoxyphenyl)-7-(1,4-dioxaspiro[4.5]dec-8-yl)-7H-pyrrolo[2,3-d]pyrimidine (150 mg, 0.30 mmol) and NH 3 -H 2 O (5 mL) in dioxane (5 mL) was heated at 120 °C for 16 h under microwave irradiation. After cooling to room temperature, the mixture was concentrated to give the title compound (170 mg, 100% yield). LC-MS: Calculated exact mass: 472.5; Observed value [M + H] + (ESI) = 473.1.

[0491] 4-(4-Amino-5-(3-methoxy-4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexanone

[0492] To a suspension of 5-(3-methoxy-4-phenoxyphenyl)-7-(1,4-dioxaspiro[4.5]dec-8-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (340 mg, 0.6 mmol) in acetone (20 mL) and THF (6 mL) was added 6N HCl (3 mL, 18 mmol). The reaction mixture was heated at 40 °C for 3 h. It was neutralized with 1N NaOH solution. Extracted with DCM, dried over sodium sulfate, evaporated and purified by column chromatography (EA) to obtain the desired product as a pale yellow solid (200 mg, 78% yield).

[0493] (S)-tert-Butyl 4-((trans)-4-(4-amino-5-(3-methoxy-4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexyl)-2-methylpiperazine-1-carboxylate

[0494] To a reaction mixture of 4-(4-amino-5-(3-methoxy-4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexanone (40 mg, 0.09 mmol), (S)-tert-butyl 2-methylpiperazine-1-carboxylate (37 mg, 0.18 mmol) and Ti(OiPr) 4 (6 drops) in DCM (10 mL) was stirred at room temperature for 30 min. NaBH(OAc) 3 (76 mg, 0.36 mmol) was added. The reaction mixture was stirred at room temperature overnight, then quenched with MeOH. Subsequently, aqueous NaHCO 3 solution and DCM were added. After filtration, the filtrate was extracted with DCM. The organic layer was collected and concentrated. The residue was purified by Prep-TLC (DCM:MeOH = 20:1) to give the product (30 mg, 54% yield). LCMS: Calculated exact mass = 612.3; experimental value [M+H] + (ESI) = 613.2.

[0495] Example 134

[0496] 5-(3-Methoxy-4-phenoxyphenyl)-7-((trans)-4-((S)-3-methylpiperazin-1-yl)cyclohexyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0497]

[0498] To a solution of (S)-tert-butyl 4-((trans)-4-(4-amino-5-(3-methoxy-4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexyl)-2-methylpiperazine-1-carboxylate (30 mg, 0.05 mmol) in DCM (10 mL) was added dropwise TFA (1 mL). The reaction mixture was stirred at room temperature for 3 h, then concentrated and the residue was purified by Prep-HPLC to afford the product as a white solid (18 mg, 70% yield). LCMS: calculated exact mass = 512.2; experimental value [M+H] + (ESI) = 513.2; 1 H NMR (600 MHz, DMSO-d 6 ) δ ppm 8.14 (s, 1H) 7.46 - 7.52 (m, 1H) 7.33 (t, J = 7.78 Hz, 2H) 7.19 - 7.23 (m, 1H) 7.10 (d, J = 8.24 Hz, 1H) 7.01 - 7.08 (m, 2H) 6.92 (d, J = 8.47 Hz, 2H) 6.22 (br.s., 1H) 4.58 (t, J = 11.79 Hz, 1H) 3.80 (s, 3H) 3.20 (d, J = 11.22 Hz, 1H) 3.12 (br.s., 1H) 2.85 - 3.00 (m, 3H) 239 (br.s., 1H) 2.25 (t, J = 10.99 Hz, 1H) 1.98 - 2.05 (m, 2H) 1.87 - 1.98 (m, 4H) 1.44 - 1.58 (m, 2H) 1.13 - 1.21 (m, 3H).

[0499] Example 135

[0500] (2S)-tert-butyl 4-(3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)cyclopentyl)-2-methylpiperazine-1-carboxylate

[0501]

[0502] 3-(4-amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-1-yl)cyclopentanol

[0503] To a 250 mL round-bottom flask, add 3-iodo-1H-pyrazolo[3,4-d]pyrimidin-4-amine (2 g, 7.64 mmol), cyclopentane-1,3-diol (3.9 g, 38.2 mmol), triphenylphosphine (6.0 g, 22.9 mmol), and THF (50 mL). Stir the mixture and add DEAD (3.98 g, 22.9 mmol) dropwise to the mixture under an ice bath. Stir the resulting solution at room temperature overnight. Filter the mixture and concentrate the filtrate under vacuum. Load the residue onto a silica gel column and elute with DCM-MeOH (100:1 - 10:1) to obtain the product as a brown solid (400 mg, 24% yield).

[0504] 3-(4-Amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-1-yl)cyclopentanone

[0505] To a 100 mL round-bottom flask, add 3-(4-amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-1-yl)cyclopentanol (316 mg, 0.92 mmol), PCC (236 mg, 1.1 mmol), and DCM (10 mL). Stir the mixture overnight. Filter the mixture and concentrate the filtrate under vacuum. Load the residue onto a silica gel column and elute with DCM-MeOH (100:1 - 20:1) to obtain the product as a white solid (100 mg, 31.8% yield).

[0506] 3-(4-Amino-3-(2-fluoro-4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)cyclopentanone

[0507] To a 100 mL round-bottom flask, add 3-(4-amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-1-yl)cyclopentanone (100 mg, 0.291 mmol), 3-(4-amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-1-yl)cyclopentanone (137 mg, 0.437 mmol), sodium carbonate (92.5 mg, 0.87 mmol), Pd(dppf)Cl 2 (42.6 mg, 0.058 mmol, 0.2 equiv) and dioxane-H 2 O (10 mL - 1 mL). Stir the resulting mixture at 85 °C overnight. Filter the mixture and concentrate the filtrate. Load the residue onto a silica gel column and elute with DCM / MeOH (50:1 - 20:1) to obtain the product (52 mg, 44% yield).

[0508] (2S)-tert-Butyl 4-(3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)cyclopentyl)-2-methylpiperazine-1-carboxylate

[0509] To a 20 mL round-bottom flask was added 3-(4-amino-3-(2-fluoro-4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)cyclopentanone (52 mg, 0.129 mmol), 1-methylpiperazine (25.8 mg, 0.258 mmol), and DCE (3 mL). The mixture was stirred overnight at room temperature. Sodium triacetoxyborohydride (41 mg, 0.193 mmol) was added to the mixture, and then the mixture was stirred at RT for 3 h. Subsequently, the mixture was filtered through celite and concentrated. The residue was loaded onto a silica column and eluted with DCM-MeOH (50:1 - 10:1), and then purified by prep-HPLC to give the product as a white solid (3.0 mg, 4.7% yield). LCMS: calculated exact mass = 487.25; experimental value [M+H] + (ESI) = 488.20; 1 H NMR(CDCl 3 ) δ ppm: 8.32 (s, 1H), 7.56 (t, J = 8.5 Hz, 1H), 7.44 (t, J = 8.0 Hz, 2H), 7.23 (s, 1H), 7.12 (d, J = 7.7 Hz, 2H), 6.96 (dd, J = 8.5, 2.2 Hz, 1H), 6.86 (dd, J = 11.4, 2.2 Hz, 1H), 5.29 - 5.43 (m, 1H), 3.13 - 3.39 (m, 8H), 2.75 (s, 3H), 2.08 - 2.56 (m, 7H).

[0510]

[0511] tert-Butyl 7-((cis)-4-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexyl)-2,7-diazaspiro[4.4]nonane-2-carboxylate

[0512] and

[0513] tert-Butyl 7-((trans)-4-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexyl)-2,7-diazaspiro[4.4]nonane-2-carboxylate

[0514] A reaction mixture of 4-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexanone (1.0 g, 2.81 mmol), tert-butyl 2,7-diazaspiro[4.4]nonane-2-carboxylate (1.9 g, 8.43 mmol), and acetic acid (101 mg, 1.68 mmol) in THF (50 mL) was stirred at room temperature for 2 h. NaBH3 CN (529 mg, 8.43 mmol). It was stirred at room temperature for 3 hours when quenched with MeOH (10 mL). Subsequently, an aqueous solution of NaHCO 3 and DCM were added, and it was filtered. The filtrate was extracted with DCM. The organic layer was collected and concentrated. The residue was purified by column chromatography on silica gel (DCM:MeOH = 10:1) to obtain tert-butyl 7-((trans)-4-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexyl)-2,7-diazaspiro[4.4]nonane-2-carboxylate (300 mg, 18% yield). LCMS: calculated exact mass = 566.2; experimental value [M+H] + (ESI) = 567.0 and tert-butyl 7-((cis)-4-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexyl)-2,7-diazaspiro[4.4]nonane-2-carboxylate (330 mg, 21% yield). LCMS: calculated exact mass = 566.2; experimental value [M+H] + (ESI) = 567.0.

[0515] Example 136

[0516] 7-((trans)-4-(2,7-Diazaspiro[4.4]non-2-yl)cyclohexyl)-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0517]

[0518] 7-((trans)-4-(4-Amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexyl)-2,7-diazaspiro[4.4]nonane-2-carboxylate

[0519] tert-Butyl 7-((trans)-4-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexyl)-2,7-diazaspiro[4.4]nonane-2-carboxylate (40 mg, 0.071 mmol), (4-phenoxyphenyl) acid (18 mg, 0.085 mmol), Pd(dppf)Cl 2 (15 mg, 0.013 mol) and Na 2 CO 3 (15 mg, 0.14 mmol) in dioxane (6 mL) and H 2The mixture in O (0.6 mL) was heated at 85 °C for 2 hours under an argon atmosphere. After cooling to room temperature, the reaction mixture was concentrated and extracted with DCM (500 mL × 4). The organic layer was concentrated and purified by Prep-TLC (DCM:MeOH = 10:1) to give the product as a pale yellow solid (10 mg, 23% yield). LCMS: calculated exact mass = 608.4; experimental value [M+H] + (ESI) = 609.1.

[0520] 7-((trans)-4-(2,7-Diazaspiro[4.4]non-2-yl)cyclohexyl)-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0521] To a solution of tert-butyl 7-((trans)-4-(4-amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexyl)-2,7-diazaspiro[4.4]nonane-2-carboxylate (20.0 mg, 0.03 mmol) in DCM (2 mL) was added dropwise TFA (1 mL). The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated and purified by Prep-HPLC to give the product as a white solid (3 mg, 17% yield). LCMS: calculated exact mass = 508.3; experimental value [M+H] + (ESI) = 509.1; 1 H NMR (400 MHz, DMSO-d 6 ) δ ppm: 9.06 (br.s., 2H) 8.25 (br.s., 1H) 7.52 (br.s., 1H) 7.39 - 7.49 (m, 5H) 7.23 (s, 2H) 7.14 - 7.21 (m, 2H) 7.07 - 7.14 (m, 6H) 6.98 (s, 1H) 5.33 (t, J = 4.63 Hz, 1H) 4.58 - 4.63 (m, 1H) 3.57 - 3.62 (m, 6H) 2.77 (d, J = 4.84 Hz, 1H) 2.63 - 2.69 (m, 1H) 2.31 - 2.37 (m, 1H) 2.26 (br.s., 1H) 2.18 (br.s., 2H) 1.89 - 2.15 (m, 13H) 1.77 (br.s., 3H) 1.41 - 1.51 (m, 2H).

[0522] Example 137

[0523] 7-((cis)-4-(2,7-Diazaspiro[4.4]non-2-yl)cyclohexyl)-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0524]

[0525] tert-Butyl 7-((cis)-4-(4-amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexyl)-2,7-diazaspiro[4.4]nonane-2-carboxylate

[0526] A mixture of tert-butyl 7-((cis)-4-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexyl)-2,7-diazaspiro[4.4]nonane-2-carboxylate (450 mg, 0.80 mmol), (4-phenoxyphenyl)boronic acid (204 mg, 0.95 mmol), Pd(dppf)Cl 2 (116 mg, 0.16 mmol), and Na 2 CO 3 (169 mg, 0.59 mmol) in dioxane (30 mL) and H 2 O (3 mL) was heated at 85 °C for 2.5 h under an argon atmosphere. After cooling to room temperature, the reaction mixture was concentrated and purified by column chromatography (DCM:MeOH = 20:1) to give the product (20 mg, 4% yield). LCMS: calculated exact mass = 608.4; experimental value [M+H] + (ESI) = 609.1.

[0527] 7-((cis)-4-(2,7-Diazaspiro[4.4]non-2-yl)cyclohexyl)-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0528] To a solution of tert-butyl 7-((cis)-4-(4-amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexyl)-2,7-diazaspiro[4.4]nonane-2-carboxylate (20 mg, 0.033 mmol) in DCM (2 mL) was added dropwise TFA (1 mL). The reaction mixture was stirred at room temperature for 0.5 h. The reaction mixture was concentrated and purified by Prep-HPLC to give the product as a white solid (2 mg, 12% yield). LCMS: calculated exact mass = 508.3; experimental value [M+H] + (ESI) = 509.1; 1 H NMR (400 MHz, DMSO-d 6)δ 10.06 (broad singlet, 2H), 8.92 (broad singlet, 2H), 8.31 (broad singlet, 1H), 7.52 (broad singlet, 1H), 7.39 - 7.50 (multiplet, 3H), 7.18 (triplet, J = 7.3 Hz, 1H), 7.04 - 7.16 (multiplet, 3H), 4.84 (broad singlet, 1H), 3.79 (broad singlet, 2H), 3.29 (broad singlet, 5H), 2.34 (broad singlet, 2H), 2.25 (broad singlet, 2H), 2.05 (doublet, J = 8.1 Hz, 4H), 1.94 (broad singlet, 3H).

[0529]

[0530] Ethyl 1-(1,4-dioxaspiro[4.5]dec-8-yl)-1H-pyrazole-4-carboxylate

[0531] To a suspension of ethyl 1H-pyrazole-4-carboxylate (1 g, 7.14 mmol) in THF (10 mL) cooled in an ice bath was added 1,4-dioxaspiro[4.5]decan-8-one (2.26 g, 14.3 mmol), PPh 3 (3.75 g, 14.3 mmol), and then DEAD (2.49 g, 14.3 mmol) was added dropwise over a period of 20 minutes. The reaction mixture was stirred overnight at room temperature. The mixture was filtered. The filtrate was concentrated in vacuo. The residue was purified by column chromatography (EA / PE, 0 to 20% gradient) to obtain the desired product as a colorless oil (1.0 g, 50% yield).

[0532] Ethyl 1-(4-oxocyclohexyl)-1H-pyrazole-4-carboxylate

[0533] To a suspension of ethyl 1-(1,4-dioxaspiro[4.5]dec-8-yl)-1H-pyrazole-4-carboxylate (1 g, 3.57 mmol) in acetone (15 mL) was added 6N HCl (15 mL, 9 mmol). The reaction mixture was heated at 50 °C for 1 hour. It was neutralized with 1N aqueous NaOH. Extracted with EA (100 mL × 2), washed with brine, dried over sodium sulfate, evaporated and purified by column chromatography (EA / PE, 0 to 30% gradient) to obtain the desired product as a colorless oil (470 mg, 56% yield).

[0534] Ethyl 1-((trans)-4-hydroxycyclohexyl)-1H-pyrazole-4-carboxylate

[0535] and

[0536] Ethyl 1-((cis)-4-hydroxycyclohexyl)-1H-pyrazole-4-carboxylate

[0537] To a stirred mixture of ethyl 1-(4-oxocyclohexyl)-1H-pyrazole-4-carboxylate (470 mg, 1.99 mmol) in MeOH (15 mL) was added NaBH 4 (91 mg, 2.39 mmol). After stirring for 1 h, the mixture was concentrated in vacuo. The residue was diluted with water (20 mL) and extracted with DCM (20 mL). The organic layer was concentrated in vacuo to afford a mixture of ethyl 1-((trans)-4-hydroxycyclohexyl)-1H-pyrazole-4-carboxylate and ethyl 1-((cis)-4-hydroxycyclohexyl)-1H-pyrazole-4-carboxylate as a colorless oil (400 mg, 84% yield).

[0538] Ethyl 1-((trans)-4-(4-amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-1-yl)cyclohexyl)-1H-pyrazole-4-carboxylate

[0539] and

[0540] Ethyl 1-((cis)-4-(4-amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-1-yl)cyclohexyl)-1H-pyrazole-4-carboxylate

[0541] To a suspension of 3-iodo-1H-pyrazolo[3,4-d]pyrimidin-4-amine (240 mg, 0.924 mmol) in THF (5 mL) cooled in an ice bath was added a mixture of ethyl 1-((trans)-4-hydroxycyclohexyl)-1H-pyrazole-4-carboxylate and ethyl 1-((cis)-4-hydroxycyclohexyl)-1H-pyrazole-4-carboxylate (220 mg, 0.924 mmol), PPh 3 (363 mg, 1.38 mmol), followed by dropwise addition of DEAD (241 mg, 1.38 mmol) over a 5 min period. The reaction mixture was stirred overnight at room temperature. The mixture was filtered. The filtrate was concentrated in vacuo. The residue was purified by column chromatography (MeOH / DCM, 0 to 10% gradient) to afford the crude product (540 mg). The crude product was purified by Prep-HPLC to give a mixture of ethyl 1-((trans)-4-(4-amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-1-yl)cyclohexyl)-1H-pyrazole-4-carboxylate and ethyl 1-((cis)-4-(4-amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-1-yl)cyclohexyl)-1H-pyrazole-4-carboxylate as a white solid (160 mg, 36% yield).

[0542] Ethyl 1-((trans)-4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)cyclohexyl)-1H-pyrazole-4-carboxylate

[0543] and

[0544] Ethyl 1 - ((cis)-4-(4 - amino - 3-(4 - phenoxyphenyl)-1H - pyrazolo[3,4 - d]pyrimidin - 1 - yl)cyclohexyl)-1H - pyrazole - 4 - carboxylate

[0545] Ethyl 1 - ((trans)-4-(4 - amino - 3 - iodo - 1H - pyrazolo[3,4 - d]pyrimidin - 1 - yl)cyclohexyl)-1H - pyrazole - 4 - carboxylate, which is a white solid, is combined with ethyl 1 - ((cis)-4-(4 - amino - 3 - iodo - 1H - pyrazolo[3,4 - d]pyrimidin - 1 - yl)cyclohexyl)-1H - pyrazole - 4 - carboxylate (50 mg, 0.104 mmol), (4 - phenoxyphenyl)boronic acid (37 mg, 0.125 mmol), Pd(dppf)Cl 2 (15 mg, 0.0208 mmol), and Na 2 CO 3 (33 mg, 0.312 mmol) in a mixture of dioxane - H 2 O (3 mL - 0.3 mL) is heated overnight at 85 °C under an inert atmosphere. After cooling to room temperature, the reaction mixture is filtered and the filtrate is concentrated in vacuo and purified by Prep - HPLC to obtain ethyl 1 - ((trans)-4-(4 - amino - 3-(4 - phenoxyphenyl)-1H - pyrazolo[3,4 - d]pyrimidin - 1 - yl)cyclohexyl)-1H - pyrazole - 4 - carboxylate (20 mg, 37% yield) as a white solid. LC - MS: Calculated exact mass: 523.2 Experimental value: [M + H] + (ESI)=524.1 and ethyl 1 - ((cis)-4-(4 - amino - 3-(4 - phenoxyphenyl)-1H - pyrazolo[3,4 - d]pyrimidin - 1 - yl)cyclohexyl)-1H - pyrazole - 4 - carboxylate (6 mg, 11% yield) as a white solid. LC - MS: Calculated exact mass: 523.2 Experimental value: [M + H] + (ESI)=524.1.

[0546] Example 138

[0547] (1 - ((trans)-4-(4 - amino - 3-(4 - phenoxyphenyl)-1H - pyrazolo[3,4 - d]pyrimidin - 1 - yl)cyclohexyl)-1H - pyrazol - 4 - yl)methanol

[0548]

[0549] To a stirred mixture of ethyl 1-((trans)-4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)cyclohexyl)-1H-pyrazole-4-carboxylate (15 mg, 0.0287 mmol) in THF (2 mL) cooled in an ice bath was added LAH (2 mg, 0.0574 mmol). The mixture was purged with N 2 After stirring for 2 h, LAH (10 mg, 0.263 mmol) was added to the mixture. After complete disappearance of the starting material as monitored by TLC, the mixture was quenched with H 2 O (2 mL) and extracted with DCM (5 mL). The organic layer was concentrated in vacuo and purified by Prep-HPLC to give the title compound as a white solid (3 mg, 21.7% yield) LC-MS: calculated exact mass: 481.2; experimental value [M+H] + (ESI) = 482.1; 1 H NMR (400 MHz, CDCl 3 ) δ 11.55 (s, 1H), 8.25 (s, 1H), 7.58 (d, J = 10.9 Hz, 3H), 7.42 (t, J = 7.8 Hz, 2H), 7.22 (t, J = 7.4 Hz, 1H), 7.16 (d, J = 7.9 Hz, 2H), 7.10 (d, J = 7.9 Hz, 2H), 6.26 (s, 1H), 5.05 (s, 1H), 4.63 (s, 2H), 4.47 (s, 1H), 2.61 (s, 2H), 2.44 (s, 2H), 2.14 (dd, J = 20.5, 9.7 Hz, 4H), 1.26 (s, 1H).

[0550] Example 139

[0551] N-(4-(4-Amino-7-(4-(5-amino-1,3-dioxolan-2-yl)cyclohexyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)phenyl)-3-phenylpropanamide

[0552]

[0553] 5-Iodo-7-(4-(methoxymethylene)cyclohexyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0554] At 0 °C for 30 min, potassium tert-butoxide (1.26 g, 11.2 mmol) in THF (10 mL) was added dropwise to a solution of (methoxymethyl)triphenylphosphonium chloride (3451 mg, 11.231 mmol) in THF (15 mL). 4-(4-Amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexan-1-one (2.0 g, 5.61 mmol) was added to the mixture and then stirred overnight at room temperature. The reaction was monitored by TLC and LCMS until the starting material was completely exhausted. It was then filtered. The filtrate was concentrated to obtain the crude product. The crude product was purified by flash column chromatography (0 - 3% MeOH / DCM) to obtain the product as a yellow solid (1.9 g, 88% yield). LCMS: Calculated exact mass = 384.04; experimental value [M+H] + (ESI) = 385.61; 1 H NMR (DMSO-d 6 ) δ ppm: 8.08 (s, 1H), 7.54 (s, 1H), 6.59 (br.s., 1H), 5.95 (s, 1H), 4.56 - 4.67 (m, 1H), 3.51 (s, 3H), 2.78 (d, J = 11.3 Hz, 1H), 2.14 - 2.24 (m, 1H), 2.00 - 2.11 (m, 1H), 1.89 (t, J = 12.1 Hz, 2H), 1.68 - 1.79 (m, 3H).

[0555] 4-(4-Amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexane-1-carbaldehyde

[0556] At room temperature, 6N HCl (20 mL, 120 mmol) was added to a solution of 5-iodo-7-(4-(methoxymethylene)cyclohexyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (1900 mg, 4.945 mmol) in THF (15 mL). The mixture was stirred at room temperature for 3 hours. It was then filtered, the filtrate was adjusted to pH = 10, and then extracted with EtOAc (30 mL × 3). The organic layer was washed with brine and dried over anhydrous Na 2 SO 4 and concentrated to obtain the crude product as a yellow solid (1.15 g, 62.8% yield). LCMS: Calculated exact mass = 307.03; experimental value [M+H] + (ESI) = 307.6; 1 H NMR (DMSO-d 6)δ ppm: 9.62 (s, 1H), 8.44 (s, 1H), 7.94 (s, 1H), 4.57 (br.s., 1H), 2.37 (br.s., 1H), 2.08 (d, J = 12.5 Hz, 2H), 1.89 - 2.01 (m, 4H), 1.41 (dd, J = 12.7, 4.1 Hz, 2H). tert-Butyl (2-(4-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexyl)-1,3-dioxolan-5-yl)carbamate

[0557] A solution of 4-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexane-1-carbaldehyde (1.1 g, 2.97 mmol), tert-butyl (1,3-dihydroxypropan-2-yl)carbamate (5.68 g, 29.7 mmol), p-toluenesulfonic acid monohydrate (5.65 g, 29.7 mmol), and Na 2 CO 3 (8.43 mg, 59.4 mmol) in chloroform (50 mL) was refluxed overnight. It was then cooled to room temperature and filtered, and the solid was washed with DCM (10 mL × 3). The filtrate was concentrated, and the crude product was purified by flash column chromatography (0.5 - 2.5% MeOH / DCM) to afford the product as a white solid (350 mg, 21.7% yield). LCMS: calculated exact mass = 543.13; experimental value [M + H] + (ESI) = 543.7.

[0558] (2-(4-(4-Amino-5-(4-(3-phenylpropanamido)phenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexyl)-1,3-dioxolan-5-yl)tert-butylcarbamate

[0559] To a 50 mL round-bottom flask was added tert-butyl (2-((trans)-4-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexyl)-1,3-dioxolan-5-yl)carbamate (150 mg, 0.27 mmol), 3-phenyl-N-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)propanamide (193 mg, 0.55 mmol), Pd(dppf)Cl 2 (101 mg, 0.13 mmol), and Na 2 CO 3(143 mg, 0.82 mmol), 1,4-dioxane containing 10% water (5 mL). The mixture was stirred overnight at 80 °C. The reaction was monitored by TLC and LCMS until the starting material was completely exhausted. The mixture was cooled to room temperature, filtered and the filtrate was concentrated. The crude product was purified by flash column chromatography (MeOH / DCM, 5 - 25%) to obtain the crude product as a yellow solid (150 mg, 80% yield). LCMS: Calculated exact mass = 640.34; experimental value [M+H] + (ESI) = 640.81.

[0560] N-(4-(4-Amino-7-(4-(5-amino-1,3-dioxan-2-yl)cyclohexyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)phenyl)-3-phenylpropanamide

[0561] To a solution of tert-butyl (2-((trans)-4-(4-amino-5-(4-(3-phenylpropanamido)phenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexyl)-1,3-dioxan-5-yl)carbamate (150 mg, 0.22 mmol) in DCM (5 mL) was added TFA (2 mL), and the mixture was stirred at room temperature for 40 min. The reaction was monitored by TLC and LCMS until the starting material was completely exhausted. Subsequently, it was concentrated, and the crude product was purified by flash column chromatography (MeOH / DCM, 1 to 5%) and Prep-HPLC (acetonitrile / water, 0.1% TFA), and freeze-dried to obtain the title compound as a white solid (55 mg, 43% yield). LCMS: Calculated exact mass = 540.28; experimental value [M+H] + (ESI) = 540.79; 1 H NMR (DMSO-d 6 ) δ ppm: 10.10 (s, 1H), 8.43 (s, 1H), 8.19 (br.s., 3H), 7.70 - 7.77 (m, 3H), 7.37 - 7.45 (m, 2H), 7.23 - 7.31 (m, 4H), 7.20 (d, J = 6.7 Hz, 1H), 4.53 - 4.67 (m, 1H), 4.46 (d, J = 5.8 Hz, 1H), 3.94 - 4.05 (m, 3H), 3.29 (br.s., 1H), 2.93 (t, J = 7.5 Hz, 2H), 2.67 (t, J = 7.6 Hz, 2H), 1.97 (d, J = 7.6 Hz, 4H), 1.80 - 1.93 (m, 2H), 1.67 (br.s., 1H), 1.30 (d, J = 11.0 Hz, 2H).

[0562] Benzyl 9-(4-amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-3-azaspiro[5.5]undecane-3-carboxylate

[0563]

[0564] Benzyl 9-oxo-3-azaspiro[5.5]undecane-3-carboxylate

[0565] A solution of 3-azaspiro[5.5]undecan-9-one hydrochloride (260 mg, 1.28 mmol) and triethylamine (0.4 mL, 3.19 mmol) in THF (40 mL) was stirred at room temperature for 10 minutes. Subsequently, the mixture was cooled to 0 °C and benzyl chloroformate (0.4 mL, 2.55 mmol) was added dropwise. After the addition, the mixture was stirred at room temperature for 12 hours. The reaction mixture was diluted with ethyl acetate (120 mL), washed with water (50 mL), dried over sodium sulfate and concentrated. The crude product was purified by flash chromatography (PE:EA = 3:1) to obtain the product as a yellow solid (350 mg, 91.1% yield). LCMS: calculated exact mass = 301.2; experimental value [M+H] + (ESI) = 302.1.

[0566] Benzyl 9-hydroxy-3-azaspiro[5.5]undecane-3-carboxylate

[0567] A stirred solution of benzyl 9-oxo-3-azaspiro[5.5]undecane-3-carboxylate (350 mg, 1.16 mmol) maintained at 0 °C in methanol (15 mL) was treated with sodium borohydride (66.3 mg, 1.74 mmol). After 10 minutes, the reaction mixture was warmed to room temperature and stirred at this temperature for an additional 1.5 hours. Subsequently, the solvent was removed under reduced pressure and the resulting residue was partitioned between water (50 mL) and dichloromethane (50 mL). The separated aqueous phase was extracted with dichloromethane (50 mL) and the combined organic fractions were then dried over sodium sulfate, filtered and concentrated under reduced pressure to give the product as a colorless oil (400 mg, quantitative). LCMS: calculated exact mass = 303.2; experimental value [M+H]+(ESI) = 304.1; 1 1H NMR (400 MHz, chloroform-d) δ ppm: 7.30 - 7.45 (m, 5H), 5.14 (s, 2H), 3.63 - 3.76 (m, 1H), 3.43 - 3.50 (m, 4H), 1.74 - 1.83 (m, 2H), 1.65 - 1.73 (m, 2H), 1.40 - 1.60 (m, 4H), 1.35 - 1.40 (m, 2H), 1.17 - 1.31 (m, 2H).

[0568] Benzyl 9-(4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-3-azaspiro[5.5]undecane-3-carboxylate

[0569] Diethyl azodicarboxylate (0.5 mL, 3.45 mmol) was added dropwise to a solution of triphenylphosphine (904.9 mg, 3.45 mmol) in tetrahydrofuran (15 mL) under ice-cooling. Subsequently, the mixture was brought to room temperature, and a solution of 4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidine (338.5 mg, 1.21 mmol) and benzyl 9-hydroxy-3-azaspiro[5.5]undecane-3-carboxylate (350 mg, 1.21 mmol) in tetrahydrofuran (10 mL) was added dropwise. After completion of the addition, the mixture was stirred at room temperature for 2 hours. The solvent was evaporated in an evaporator. The residue was purified by flash chromatography (PE:EA = 3:1) to obtain the product as a yellow solid (260 mg, 39.9% yield). LCMS: calculated exact mass = 564.1; experimental value [M+H]+ (ESI) = 564.9.

[0570] Benzyl 9-(4-chloro-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-3-azaspiro[5.5]undecane-3-carboxylate

[0571] A solution of benzyl 9-(4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-3-azaspiro[5.5]undecane-3-carboxylate (240 mg, 0.43 mmol), (4-phenoxyphenyl)boronic acid (182.8 mg, 0.85 mmol), Pd(dppf)Cl 2 (30.3 mg, 0.04 mmol) and sodium carbonate (135.8 mg, 1.28 mmol) in dioxane-water (27.5 mL, 10:1) was stirred at 80 °C under a nitrogen atmosphere for 4 hours. After cooling to room temperature, the reaction mixture was diluted with ethyl acetate (120 mL), washed with water (200 mL) and brine (50 mL), dried over sodium sulfate, and concentrated. The crude product was purified by flash column chromatography (PE:EA = 3:1) to obtain the product as a yellow solid (220 mg, 85.3% yield). LCMS: calculated exact mass = 606.2; experimental value [M+H]+ (ESI) = 607.2.

[0572] Benzyl 9-(4-amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-3-azaspiro[5.5]undecane-3-carboxylate

[0573] A mixture of benzyl 9-(4-chloro-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-3-azaspiro[5.5]undecane-3-carboxylate (100 mg, 0.16 mmol), 1,4-dioxane (1.5 mL), and concentrated aqueous ammonia solution (1.5 mL) was reacted in a microwave reactor at 120 °C for 9 h. The mixture was cooled to room temperature, and the solvent was distilled off under reduced pressure and the residue was purified by Prep-TLC (DCM:MeOH = 15:1) to afford the title compound as a yellow solid (70 mg, 72.8% yield).

[0574] LCMS: Calculated exact mass = 587.3; found [M+H]+ (ESI) = 588.2.

[0575] Example 140

[0576] 4-(9-(4-Amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-3-azaspiro[5.5]undecan-3-yl)butan-1-ol

[0577]

[0578] 4-(9-(4-Amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-3-azaspiro[5.5]undecan-3-yl)butan-1-ol

[0579] A mixture of benzyl 9-(4-amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-3-azaspiro[5.5]undecane-3-carboxylate (70 mg, 0.12 mmol) and Pd / C (70 mg, 1.0 equiv w / w) in tetrahydrofuran (10 mL) was stirred at room temperature under a hydrogen atmosphere for 12 h. Subsequently, the solution was filtered and concentrated and purified by Prep-TLC (DCM:MeOH = 15:1) to afford the product as a yellow solid (23 mg, 36.7% yield). LCMS: Calculated exact mass = 525.3; found [M+H]+ (ESI) = 526.2; 11H NMR (400 MHz, chloroform-d) δ ppm: 8.30 (s, 1H), 7.33 - 7.53 (m, 4H), 6.94 - 7.24 (m, 6H), 5.25 (br.s., 2H), 4.66 (t, J = 11.6 Hz, 1H), 3.64 (t, J = 5.2 Hz, 2H), 2.78 (br.s., 4H), 2.69 (t, J = 6.0 Hz, 2H), 1.77 - 2.05 (m, 10H), 1.61 - 1.76 (m, 4H), 1.44 (t, J = 12.1 Hz, 2H).

[0580] Example 141

[0581] 5-(4-Phenoxyphenyl)-7-(3-azaspiro[5.5]undec-9-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0582]

[0583] 5-(4-Phenoxyphenyl)-7-(3-azaspiro[5.5]undec-9-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0584] A solution of benzyl 9-(4-amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-3-azaspiro[5.5]undecane-3-carboxylate (50 mg, 0.09 mmol) in dichloromethane (10 mL) was cooled to 0 °C. Subsequently, boron tribromide (1.0 mL, 1.0 M in dichloromethane) was added dropwise. After the addition, the mixture was stirred at 0 °C for an additional 0.5 h. The reaction was quenched with methanol (2 mL) and concentrated to afford the product as a yellow solid (36 mg, 93.2% yield) by Prep-TLC (DCM:MeOH: 15:1). LCMS: calculated exact mass = 453.3; found [M+H]+ (ESI) = 454.2; 1 1H NMR (400 MHz, DMSO-d 6 ) δ ppm: 8.47 (br.s., 2H), 8.14 (s, 1H), 7.56 (s, 1H), 7.37 - 7.51 (m, 4H), 7.05 - 7.22 (m, 5H), 6.14 (br.s., 2H), 4.51 - 4.66 (m, 1H), 3.08 (br.s., 4H), 1.96 - 2.14 (m, 2H), 1.82 - 1.93 (m, 4H), 1.75 (d, J = 9.9 Hz, 2H), 1.54 (br.s., 2H), 1.32 - 1.46 (m, 2H).

[0585] Example 142

[0586] 7-(3-Methyl-3-azaspiro[5.5]undecan-9-yl)-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0587]

[0588] 7-(3-Methyl-3-azaspiro[5.5]undecan-9-yl)-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0589] A solution of 5-(4-phenoxyphenyl)-7-(3-azaspiro[5.5]undecan-9-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (20 mg, 0.04 mmol), formaldehyde (7.9 mg, 0.26 mmol), acetic acid (2.4 mg, 0.04 mmol) and molecular sieve (300 mg) in dichloroethane / methanol (5 mL / 2 mL) was stirred at room temperature for 1 hour. Subsequently, sodium triacetoxyborohydride (55.1 mg, 0.26 mmol) was added and stirring was continued at room temperature for 16 hours. Water (50 mL) and dichloromethane (50 mL) were added to the reaction mixture, saturated aqueous solution of sodium bicarbonate (25 mL) was further added thereto and the mixture was partitioned. The organic layer was washed with brine (25 mL), dried over sodium sulfate and concentrated to be purified by Prep-TLC (DCM:MeOH = 15:1) to obtain the title compound as a white solid (9 mg, 43.7% yield). LCMS: calculated exact mass = 467.3; experimental value [M+H]+ (ESI) = 468.2; 1 H NMR (400 MHz, DMSO-d 6 ) δ ppm: 8.13 (s, 1H), 7.56 - 7.61 (m, 1H), 7.37 - 7.52 (m, 4H), 7.04 - 7.22 (m, 5H), 6.13 (br.s., 1H), 4.59 (t, J = 12.1 Hz, 1H), 2.95 - 3.20 (m, 4H), 2.71 (s, 3H), 2.01 (d, J = 12.2 Hz, 2H), 1.75 (d, J = 11.0 Hz, 2H).

[0590] Example 143

[0591] 2-(4-(4-Amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexyl)-4-methylmorpholin-3-one

[0592] and

[0593] Example 144

[0594] 7-(4-(4-Methylmorpholin-2-yl)cyclohexyl)-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0595]

[0596] 2-(8-Hydroxy-1,4-dioxaspiro[4.5]decan-8-yl)-4-methylmorpholin-3-one

[0597] At -78 °C under N 2 atmosphere, LDA (2 mmol / l, 19.6 mmol, 9.8 M) was added to a solution of 4-methylmorpholin-3-one (1.88 g, 16.3 mmol) in THF (20 mL). The reaction mixture was then stirred at -78 °C for 60 minutes. Thereafter, 1,4-dioxaspiro[4.5]decan-8-one (92.5 g, 16.3 mmol) in THF (10 mL) was added dropwise at -78 °C. The reaction mixture was slowly warmed to room temperature and stirred at 20 °C for 2 hours. By LC-MS and TLC detection, the starting materials were exhausted. It was quenched with saturated NH 4 Cl solution (20 mL) and extracted with EA (30 mL × 3). The organic layer was washed with brine (40 mL), dried over Na 2 SO 4 and concentrated in vacuo. The crude product was purified by flash column chromatography eluting with DCM-MeOH (200 / 1 - 50 / 1) to give the desired product as a yellow solid (2.8 g, 64%, yield). LC-MS: calculated exact mass = 271.14; experimental value [M+H] + (ESI) = 272.1; 1 H NMR (400 MHz, chloroform-d) δ ppm: 4.06 (dd, J = 4.16, 11.69 Hz, 1H), 3.95 - 4.01 (m, 4H), 3.93 (s, 1H), 3.75 - 3.84 (m, 1H), 3.65 (dt, J = 4.43, 11.75 Hz, 1H), 3.13 (dd, J = 2.82, 11.95 Hz, 1H), 1.97 - 2.20 (m, 3H), 1.88 (dt, J = 4.03, 13.43 Hz, 1H), 1.54 - 1.65 (m, 3H), 1.42 - 1.50 (m, 1H).

[0598] 4-Methyl-2-(4-oxocyclohexylidene)morpholin-3-one

[0599] At 0 °C under N 2In an atmosphere, POCl was added to a solution of 2-(8-hydroxy-1,4-dioxaspiro[4.5]dec-8-yl)-4-methylmorpholin-3-one (2.3 g, 8.5 mmol) and TEA (17.2 g, 170 mmol) in DCM (30 mL). 3 (13.0 g, 85 mmol). Subsequently, the reaction mixture was stirred at 20 °C for 6 hours. The reaction was monitored by LC-MS and TLC until the starting materials were exhausted. The reaction mixture was quenched with ice water (20 mL) and extracted with DCM (30 mL × 3). The organic layer was washed with brine (50 mL), dried over Na 2 SO 4 and concentrated in vacuo. The crude product was purified by flash column chromatography eluting with PE-EA (5 / 1 - 1 / 1) to afford the product as a colorless oil (1.0 g, 57%, yield). LC-MS: calculated exact mass = 209.11; experimental value [M+H] + (ESI) = 210.1; 1 1H NMR (400 MHz, chloroform-d) δ ppm: 4.08 (t, J = 5.04 Hz, 2H), 3.50 (t, J = 5.04 Hz, 2H), 3.32 (t, J = 6.87 Hz, 2H), 3.00 - 3.09 (m, 3H), 2.72 (t, J = 6.87 Hz, 2H), 2.37 - 2.49 (m, 4H).

[0600] 4-Methyl-2-(4-oxocyclohexyl)morpholin-3-one

[0601] To a solution of 4-methyl-2-(4-oxocyclohexylidene)morpholin-3-one (84 mg, 0.3 mmol) in THF (15 mL) was added Pd / C (100 mg, 10%). Subsequently, the reaction mixture was hydrogenated at 20 °C under 30 psi for 4 hours. By LC-MS and TLC detection, the starting materials were exhausted. It was filtered and concentrated in vacuo to afford the desired product as a pale yellow oil (1.0 g, 98% yield). LC-MS: calculated exact mass = 211.12; experimental value [M+H] + (ESI) = 212.1; 1HNMR (400 MHz, chloroform-d) δ ppm: 4.11 (d, J = 1.88 Hz, 1H), 4.02 (dd, J = 3.76, 11.82 Hz, 1H), 3.78 (dt, J = 3.09, 11.62 Hz, 1H), 3.58 - 3.71 (m, 1H), 3.13 (dd, J = 2.55, 11.95 Hz, 1H), 2.99 - 3.05 (m, 3H), 2.57 - 2.70 (m, 1H), 2.31 - 2.48 (m, 4H), 1.73 - 2.05 (m, 4H).

[0602] 2-(4-Hydroxycyclohexyl)-4-methylmorpholin-3-one

[0603] At 0 °C under N 2 atmosphere, NaBH 4 (380 mg, 10 mmol) was added to a solution of compound 4-methyl-2-(4-oxocyclohexyl)morpholin-3-one (0.5 g, 2.5 mmol) in THF (20 mL). Subsequently, the reaction mixture was stirred at 20 °C for 2 h. The reaction was monitored by LC-MS and TLC until the starting material was exhausted. It was quenched with saturated NH 4 Cl solution (20 mL) and extracted with DCM (30 mL × 5). The organic layer was washed with brine (20 mL), dried over Na 2 SO 4 and concentrated. The crude product was purified by flash column chromatography eluting with PE-EA (10 / 1 - 1 / 5) to give the product as a colorless oil (126 mg, 24% yield). LC-MS: calculated exact mass = 213.14; experimental value [M + H] + (ESI) = 214.1; 1 H NMR (400 MHz, chloroform-d) δ ppm: 3.97 - 4.02 (m, 2H), 3.76 (dt, J = 2.55, 11.48 Hz, 1H), 3.45 - 3.67 (m, 2H), 3.10 (d, J = 11.28 Hz, 1H), 3.00 (s, 3H), 1.93 - 2.18 (m, 3H), 1.69 - 1.76 (m, 1H), 1.22 - 1.57 (m, 5H).

[0604] 2-(4-(4-Chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexyl)-4-methylmorpholin-3-one

[0605] At 0 °C under N 2Under an atmosphere, DIAD (182 mg, 0.9 mmol) was added dropwise to a solution of 2-(4-hydroxycyclohexyl)-4-methylmorpholin-3-one (84 mg, 0.3 mmol), 4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidine (63 mg, 0.3 mmol), and PPh 3 (236 mg, 0.9 mmol) in THF (12 mL). Subsequently, the reaction mixture was stirred at 20 °C for 16 h. Monitoring by LC-MS and TLC indicated near exhaustion of the starting materials. The reaction mixture was concentrated and purified by Prep-TLC eluting with PE-EA (3:1) to afford the desired product as a yellow solid (120 mg, 80% yield). LC-MS: calculated exact mass = 474.03; found [M+H] + (ESI) = 475.0.

[0606] 2-(4-(4-chloro-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexyl)-4-methylmorpholin-3-one

[0607] To a suspension of compound 2-(4-(4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexyl)-4-methylmorpholin-3-one (150 mg, 0.3 mmol) and (4-phenoxyphenyl)boronic acid (67 mg, 0.3 mmol), Na 2 CO 3 (102 mg, 0.9 mmol), and Pd(dppf)Cl 2 (22 mg, 0.03 mmol) in THF and water (15 mL - 3 mL) was added N 2 2 and the mixture was stirred at 65 °C for 4 h. Monitoring by LC-MS and TLC indicated near exhaustion of the starting materials. The reaction mixture was concentrated and purified by flash column chromatography eluting with DCM-MeOH (100 / 1 - 20 / 1) to afford the desired product as a yellow solid (130 mg, 84% yield).

[0608] LC-MS: calculated exact mass = 516.19; found [M+H] + (ESI) = 517.2; 11H NMR (400 MHz, chloroform-d) δ ppm: 8.63 (s, 1H), 7.63 - 7.71 (m, 3H), 7.52 - 7.59 (m, 2H), 7.43 - 7.51 (m, 6H), 7.33 - 7.41 (m, 3H), 6.95 - 7.18 (m, 6H), 4.94 - 5.04 (m, 1H), 4.17 (d, J = 4.03 Hz, 1H), 4.04 (td, J = 2.08, 9.81 Hz, 1H), 3.77 (dt, J = 3.22, 11.15 Hz, 1H), 3.61 (dt, J = 4.30, 11.28 Hz, 1H), 3.16 (d, J = 11.82 Hz, 1H), 3.00 (s, 3H), 2.45 (d, J = 4.57 Hz, 1H), 2.18 - 2.35 (m, 2H), 1.91 - 2.09 (m, 3H), 1.77 - 1.90 (m, 2H).

[0609] 2-(4-(4-Amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexyl)-4-methylmorpholin-3-one

[0610] To a solution of the compound 2-(4-(4-chloro-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexyl)-4-methylmorpholin-3-one (130 mg, 0.25 mmol) in 1,4-dioxane (5 mL) in a sealed tube was added NH 3 .H 2 O (8 mL). The reaction mixture was then stirred at 120 °C for 48 h. By LC-MS and TLC detection, the starting material was nearly exhausted. Extracted with EA (30 mL × 3), washed with brine (40 mL), dried over Na 2 SO 4 , concentrated in vacuo and purified by pre-TLC eluting with DCM / MeOH (20 / 1) to give the desired product as a yellow solid (100 mg, 80% yield). LC-MS: calculated exact mass = 497.24; experimental value [M + H] + (ESI) = 498.2; 1 1H NMR (400 MHz, methanol-d 4)δ ppm: 8.29 (s, 1H), 7.57 (s, 1H), 7.47 (d, J = 8.60 Hz, 2H), 7.32 - 7.40 (m, 2H), 7.02 - 7.16 (m, 5H), 4.21 (d, J = 4.57 Hz, 1H), 3.98 - 4.10 (m, 1H), 3.70 - 3.84 (m, 1H), 3.52 - 3.66 (m, 1H), 3.29 - 3.31 (m, 1H), 3.22 (d, J = 12.09 Hz, 1H), 2.95 (s, 3H), 2.21 - 2.43 (m, 3H), 1.74 - 2.08 (m, 5H), 1.49 - 1.65 (m, 1H).

[0611] 7-(4-(4-Methylmorpholin-2-yl)cyclohexyl)-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0612] At 0 °C under N 2 atmosphere, LAH (2 mL) was added to a solution of 2-(4-(4-amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclohexyl)-4-methylmorpholin-3-one (50 mg, 0.1 mmol) in THF (5 mL). The reaction mixture was then stirred at 20 °C for 2 h. Monitored by LC-MS and TLC, the starting material was exhausted. The reaction mixture was quenched successively with saturated water (0.076 mL), NaOH (0.076 mL, 15%), and water (0.228 mL). Subsequently, Na 2 SO 4 was added and the mixture was stirred for 30 min. The mixture was filtered through diatomaceous earth and washed with MeOH (20 mL). The filtrate was concentrated and purified by Prep-HPLC to give the desired product as a white solid (10 mg, 10% yield). LC-MS: calculated exact mass = 483.26; experimental value [M + H] + (ESI) = 484.3; 1 H NMR (600 MHz, methanol-d 4)δ ppm: 8.30 (s, 1H), 7.59 (s, 1H), 7.49 (d, J = 8.59 Hz, 2H), 7.39 (t, J = 8.01 Hz, 2H), 7.16 (t, J = 7.43 Hz, 1H), 7.12 (d, J = 8.59 Hz, 2H), 7.07 (d, J = 7.76 Hz, 2H), 4.15 (d, J = 11.06 Hz, 1H), 4.02 - 4.10 (m, 1H), 3.87 (t, J = 12.39 Hz, 1H), 3.70 (d, J = 12.06 Hz, 1H), 3.45 (d, J = 12.22 Hz, 1H), 3.32 - 3.33 (m, 1H), 3.09 (t, J = 10.82 Hz, 1H), 2.94 (s, 3H), 2.87 (t, J = 11.48 Hz, 1H), 2.08 - 2.25 (m, 3H), 1.93 (dt, J = 4.38, 8.30 Hz, 2H), 1.80 - 1.87 (m, 3H), 1.65 - 1.76 (m, 1H).

[0613]

[0614] 2-Hydroxypropane-1,3-diyl dibenzoate

[0615] At 0 °C, NaBH 4 (352 mg, 9.6 mmol) was added to a solution of 2-oxopropane-1,3-diyl dibenzoate (2.5 g, 8.4 mmol) in THF (50 mL), and the mixture was stirred at 0 °C for 10 minutes. The reaction was monitored by TLC until the starting material was exhausted. It was quenched with NH 4 Cl solution and extracted with EA. The organic layer was washed with brine (40 mL), dried over Na 2 SO 4 and concentrated. The crude product was purified by flash column chromatography (PE:EA = 10:1 - 3:1) to obtain the desired product as a colorless oil (1.8 g, 75.6% yield). 1 1H NMR (400 MHz, DMSO-d 6 ) δ ppm: 8.00 (d, J = 7.25 Hz, 4H), 7.61 - 7.73 (m, 2H), 7.45 - 7.56 (m, 4H), 5.57 (d, J = 5.64 Hz, 1H), 4.32 - 4.44 (m, 4H), 4.15 - 4.25 (m, 1H).

[0616] 2-(4-Chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)propane-1,3-diyl dibenzoate

[0617] At 0 °C in N2 Under an atmosphere, DEAD (2.61 g, 15 mmol) was added dropwise to a solution of 2-hydroxypropane-1,3-diyl dibenzoate (1.4 g, 5 mmol), 4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidine (1.8 g, 6 mmol), and PPh 3 (3.93 g, 15 mmol) in THF (40 mL). Subsequently, the reaction mixture was stirred at 20 °C for 18 h. The reaction mixture was concentrated and the crude product was purified by Prep-TLC eluting with PE / EA (100 / 1 - 3 / 1) to give the desired product as a white solid (2 g, 71% yield). LC-MS: Calculated exact mass = 561.00; Experimental value [M+H] + (ESI) = 562.0. 2-(4-chloro-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)propane-1,3-diyl dibenzoate

[0618] 2-(4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)propane-1,3-diyl dibenzoate (2.2 g, 3.9 mmol)), (4-phenoxyphenyl)boronic acid (1.0 g, 4.7 mmol), Na 2 CO 3 (2.48 g, 23.4 mmol), and Pd(dppf)Cl 2 (571 mg, 0.78 mmol) in a suspension in THF and water (30 mL - 6 mL) were stirred at 65 °C under N 2 atmosphere for 3 h. The reaction mixture was concentrated and the crude product was purified by flash column chromatography (PE:EA = 20:1 - 3:1) to give the desired product as a yellow solid (2.0 g, 85% yield). LC-MS: Calculated exact mass = 603.16; Experimental value [M+H] + (ESI) = 604.2.

[0619] 2-(4-amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)propane-1,3-diol

[0620] In a sealed tube, NH 3 .H 2 O (30 mL) was added to a solution of 2-(4-chloro-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)propane-1,3-diyl dibenzoate (2 g, 3.3 mmol) in 1,4-dioxane (30 mL). Subsequently, the reaction mixture was stirred at 120 °C for 60 h. The reaction mixture was extracted with EA (30 mL×3). The organic layer was washed with brine (40 mL), over Na2 SO 4 Dry and concentrated. The crude product was purified by Prep-TLC (DCM:MeOH = 100:1 - 10:1) to give the desired product as a yellow solid (750 mg, 60% yield).

[0621] LC-MS: Calculated exact mass = 376.15; Experimental value [M+H] + (ESI) = 377.2; 1 H NMR (400 MHz, DMSO-d 6 ) δ ppm: 8.13 (s, 1H), 7.47 (d, J = 8.53 Hz, 2H), 7.41 (t, J = 7.91 Hz, 2H), 7.32 (s, 1H), 7.16 (t, J = 7.40 Hz, 1H), 7.07 - 7.13 (m, 4H), 5.93 (br.s., 2H), 4.69 - 4.90 (m, 3H), 3.84 (br.s., 4H).

[0622] 5-(4-Phenoxyphenyl)-7-((2r,5r)-2-(piperidin-4-yl)-1,3-dioxolan-5-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine and 5-(4-phenoxyphenyl)-7-((2s,5s)-2-(piperidin-4-yl)-1,3-dioxolan-5-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0623] A suspension of 2-(4-amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)propane-1,3-diol (50 mg, 0.13 mmol), tert-butyl 4-formylpiperidine-1-carboxylate (360 mg, 1.69 mmol), TSOH.H 2 O (321 mg, 1.69 mmol) and Na 2 SO 4 (1.85 g, 13 mmol) in CHCl 3 (15 mL) was stirred under reflux in an N 2 atmosphere for 48 h. The reaction mixture was quenched with Na 2 CO 3 solution (40 mL) and extracted with EA (40 mL × 3). The organic layer was washed with brine (40 mL), dried over Na 2 SO 4Dry and concentrated. The crude product was purified by Prep-TLC (PE:EA = 10:1) and Prep-HPLC to obtain 5-(4-phenoxyphenyl)-7-((2r,5r)-2-(piperidin-4-yl)-1,3-dioxan-5-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (10 mg, 16% yield), LC-MS: calculated exact mass = 471.23; experimental value [M+H] + (ESI)=472.1; 1 H NMR(600MHz, methanol-d 4 )δppm 8.26(s,1H),7.84(s,1H),7.50(d,J = 8.47Hz,2H),7.40(t,J = 7.90Hz,2H),7.17(t,J = 7.44Hz,1H),7.13(d,J = 8.47Hz,2H),7.08(d,J = 8.01Hz,2H),4.83 - 4.84(m,1H),4.74(d,J = 4.58Hz,1H),4.38 - 4.46(m,2H),4.31(d,J = 12.59Hz,2H),3.41(d,J = 12.36Hz,2H),2.98(t,J = 12.02Hz,2H),2.01 - 2.08(m,3H),1.57 - 1.72(m,2H); and 5-(4-phenoxyphenyl)-7-((2s,5s)-2-(piperidin-4-yl)-1,3-dioxan-5-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (5 mg, 8% yield), LC-MS: calculated exact mass = 603.16; experimental value [M+H] + (ESI)=472.1; 1 H NMR(600MHz, methanol-d 4) δ ppm 8.34 (s, 1H), 7.55 (s, 1H), 7.48 (d, J = 8.47 Hz, 2H), 7.39 (t, J = 7.90 Hz, 2H), 7.16 (t, J = 7.32 Hz, 1H), 7.12 (d, J = 8.47 Hz, 2H), 7.07 (d, J = 8.01 Hz, 2H), 5.08 (td, J = 5.52, 10.70 Hz, 1H), 4.65 (d, J = 4.35 Hz, 1H), 4.30 - 4.35 (m, 2H), 4.22 - 4.29 (m, 2H), 3.43 (d, J = 12.59 Hz, 2H), 2.96 - 3.06 (m, 2H), 1.95 - 2.12 (m, 3H), 1.62 - 1.74 (m, 2H); and tert-butyl 4-(5-(4-amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-1,3-dioxolan-2-yl)piperidine-1-carboxylate.

[0624] Example 145

[0625] 7-((2r,5r)-2-(1-methylpiperidin-4-yl)-1,3-dioxolan-5-yl)-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0626] and

[0627] Example 146

[0628] 7-((2s,5s)-2-(1-methylpiperidin-4-yl)-1,3-dioxolan-5-yl)-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0629]

[0630] At 0 °C under N 2 atmosphere, LAH (2 mL, 2 mmol) was added to a solution of tert-butyl 4-(5-(4-amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-1,3-dioxolan-2-yl)piperidine-1-carboxylate (130 mg, 0.28 mmol) in THF (5 mL). The reaction mixture was then stirred at 20 °C for 16 h. The reaction was quenched by the sequential addition of saturated aqueous (0.076 mL), NaOH (0.076 mL, 15%), and water (0.228 mL). Subsequently, Na 2 SO 4And stir the mixture for 30 min. Filter it through diatomaceous earth and wash with MeOH. Concentrate the filtrate. Purify the crude product by Prep-HPLC to obtain 7-((2r,5r)-2-(1-methylpiperidin-4-yl)-1,3-dioxan-5-yl)-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (15 mg, 11% yield), LC-MS: calculated exact mass = 471.23; experimental value [M+H] + (ESI) = 486.2; 1 H NMR(600MHz, methanol-d 4 )ppm: 8.33(s, 1H), 7.92(s, 1H), 7.50(d, J = 8.47Hz, 2H), 7.40(t, J = 7.90Hz, 2H), 7.17(t, J = 7.44Hz, 1H), 7.14(d, J = 8.47Hz, 2H), 7.09(d, J = 8.24Hz, 2H), 4.92(br.s., 1H), 4.75(d, J = 4.58Hz, 1H), 4.42(d, J = 12.36Hz, 2H), 4.31(d, J = 12.36Hz, 2H), 3.53(d, J = 12.36Hz, 2H), 2.97(t, J = 12.13Hz, 2H), 2.83(s, 3H), 2.08(d, J = 13.96Hz, 2H), 1.98(ddd, J = 3.89, 8.35, 16.14Hz, 1H), 1.65 - 1.75(m, 2H); and 7-((2s,5s)-2-(1-methylpiperidin-4-yl)-1,3-dioxan-5-yl)-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (10 mg, 7%), LC-MS: calculated exact mass = 471.23; experimental value [M+H] + (ESI) = 486.2; 1 H NMR(600MHz, methanol-d 4)ppm: 8.34 (s, 1H), 7.56 (s, 1H), 7.48 (d, J = 8.47 Hz, 2H), 7.39 (t, J = 7.78 Hz, 2H), 7.17 (t, J = 7.44 Hz, 1H), 7.12 (d, J = 8.47 Hz, 2H), 7.07 (d, J = 8.24 Hz, 2H), 5.05 - 5.14 (m, 1H), 4.66 (d, J = 4.35 Hz, 1H), 4.30 - 4.35 (m, 2H), 4.23 - 4.29 (m, 2H), 3.55 (d, J = 12.36 Hz, 2H), 3.00 (t, J = 12.13 Hz, 2H), 2.86 (s, 3H), 2.09 (d, J = 14.42 Hz, 2H), 1.97 (tdd, J = 3.98, 8.13, 12.07 Hz, 1H), 1.66 - 1.77 (m, 2H).

[0631] 5-Iodo-7-(piperidin-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0632]

[0633] tert-Butyl 4-(4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)piperidine-1-carboxylate

[0634] To a suspension of 4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidine (50 g, 179 mmol) in THF (900 mL) cooled in an ice bath was added tert-butyl 4-hydroxypiperidine-1-carboxylate (54.1 g, 269 mmol), PPh 3 (78 g, 448 mmol), and then DEAD (117.4 g, 448 mmol) was added dropwise over a period of 60 minutes. The reaction mixture was stirred overnight at room temperature. The mixture was evaporated. The residue was diluted with EA (900 mL). The solid thus obtained was collected by filtration. Dried under reduced pressure to give the title compound (35 g, 42% yield).

[0635] tert-Butyl 4-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)piperidine-1-carboxylate

[0636] tert-Butyl 4-(4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)piperidine-1-carboxylate (25 g, 54.1 mmol) was placed in dioxane (200 mL) NH 3 -H 2The mixture in O (200 mL) was reacted in a sealed container at 120 °C for 8 hours. After cooling to room temperature, the reaction mixture was concentrated to give the product as a pale yellow solid (20 g, 84% yield). LCMS: calculated exact mass = 443.1; experimental value [M+H] + (ESI) = 443.6.

[0637] 5-Iodo-7-(piperidin-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0638] To a solution of tert-butyl 4-(4-amino-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)piperidine-1-carboxylate (15 g, 33.9 mmol) in DCM (80 mL) was added dropwise TFA (50 mL). The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated, and 1 M sodium hydroxide was added to adjust to pH 8. The solid formed was filtered to give the product as a white solid (12 g, 100% yield). LCMS: calculated exact mass = 343.0; experimental value [M+H] + (ESI) = 343.6.

[0639] 3-Iodo-1-(piperidin-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine

[0640] Method-1

[0641]

[0642] tert-Butyl 4-(4-amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidine-1-carboxylate

[0643] To an ice-bath cooled suspension of 3-iodo-1H-pyrazolo[3,4-d]pyrimidin-4-amine (20.0 g, 76.6 mmol) in THF (600 mL) was added tert-butyl 4-hydroxypiperidine-1-carboxylate (38.5 g, 191.6 mmol), PPh 3 (50.2 mg, 191.6 mmol), and then DEAD (33.4 g, 191.6 mmol) was added dropwise over a period of 1 hour. The reaction was stirred at room temperature for 3 hours and monitored by LCMS until the starting material was completely converted. The reaction mixture was concentrated. The crude product was passed through THF (100 mL), and then washed and filtered through EtOAc (250 mL) to give the product (11.1 g, 32% yield). LCMS: calculated exact mass = 444.1; experimental value [M+H] + (ESI) = 445.0.

[0644] 3-Iodo-1-(piperidin-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine

[0645] To a solution of tert-butyl 4-(4-amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidine-1-carboxylate (11.1 g, 25.0 mmol) in DCM (100 mL) was added dropwise TFA (25 mL). The reaction mixture was stirred at room temperature for 3 h. The reaction mixture was concentrated and purified by column chromatography on silica (DCM:MeOH = 10:1) to afford the product (7.5 g, 87% yield). LCMS: calculated exact mass = 344.0; experimental value [M+H] + (ESI) = 344.9.

[0646] Method-2

[0647]

[0648] tert-Butyl 4-(4-chloro-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidine-1-carboxylate

[0649] Diethyl azodicarboxylate (16.8 mL, 106.97 mmol) was added dropwise to a solution of triphenylphosphine (28.1 g, 106.97 mmol) in THF (200 mL) under ice-cooling. Subsequently, the mixture was allowed to reach room temperature, and a solution of 4-chloro-3-iodo-1H-pyrazolo[3,4-d]pyrimidine (10 g, 35.66 mmol) and tert-butyl 4-hydroxypiperidine-1-carboxylate (9.3 g, 46.35 mmol) in THF / DMSO (150 mL / 50 mL) was added dropwise thereto. After completion of the dropwise addition, the mixture was stirred at room temperature for 2 h. The mixture was diluted with ethyl acetate (200 mL), washed with water (350 mL) and brine (300 mL), dried over sodium sulfate and concentrated to be purified by flash chromatography (PE:EA = 3:1) to afford tert-butyl 4-(4-chloro-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidine-1-carboxylate as a pink solid (2.4 g, 14% yield). LCMS: calculated exact mass = 463.0; experimental value [M+H] + (ESI) = 463.6.

[0650] tert-Butyl 4-(4-amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidine-1-carboxylate

[0651] A mixture of tert-butyl 4-(4-chloro-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidine-1-carboxylate (2.4 g, 5.18 mmol), 1,4-dioxane (35 mL) and concentrated aqueous ammonia solution (35 mL) was heated in a pressure vessel at 120 °C for 16 h. After cooling to room temperature, the mixture was concentrated in vacuo. The residue was suspended in water (20 mL), filtered and washed with water (10 mL) to afford a white solid (2.0 g, 87% yield). LCMS: calculated exact mass = 444.1; found [M+H]+ (ESI) = 444.6.

[0652] 3-Iodo-1-(piperidin-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine

[0653] A solution of tert-butyl 4-(4-amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-1-yl)piperidine-1-carboxylate (2.0 g, 4.50 mmol) in TFA-DCM (4 mL - 4 mL) was stirred at room temperature for 0.5 h. The mixture was concentrated to give the crude product, which was used in the next step without further purification. LCMS: calculated exact mass = 344.0; found [M+H] + (ESI) = 344.6.

[0654] 5-(4-Phenoxyphenyl)-7-(piperidin-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0655]

[0656] tert-Butyl 4-(4-chloro-5-iodo-7H-pyrrolo-[2,3-d]pyridin-7-yl)piperidine-1-carboxylate

[0657] To a suspension of 4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidine (10.0 g, 35.8 mmol) in THF (250 mL) cooled in an ice bath was added tert-butyl 4-hydroxypiperidine-1-carboxylate (11 g, 53.7 mmol), PPh 3 (21 mg, 82.3 mmol), followed by dropwise addition of DEAD (14.3 g, 82.3 mmol) over a period of 1 h. The reaction mixture was stirred at room temperature for 3 h and monitored by LCMS until complete conversion of the starting material. The reaction mixture was concentrated. The crude product was washed with THF (100 mL) and then with EtOAc (250 mL) to afford the product (9.3 g, 37% yield). LCMS: calculated exact mass = 462.0; found [M+H] + (ESI) = 463.1.

[0658] tert-Butyl 4-(4-chloro-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)piperidine-1-carboxylate

[0659] A mixture of tert-butyl 4-(4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidin-7-yl)piperidine-1-carboxylate (8.9 g, 19.26 mmol), (4-phenoxyphenyl)boronic acid (8.2 g, 38.52 mmol), Pd(dppf)Cl 2 (1.4 g, 1.93 mmol), and Na 2 CO 3 (4.1 g, 38.52 mmol) in dioxane (150 mL) and H 2 O (15 mL) was heated at 85 °C under an argon atmosphere for 3 hours. After cooling to room temperature, the reaction mixture was concentrated and extracted with DCM (500 mL × 4). The organic layer was concentrated and purified by flash column chromatography (PE:EA = 6:1) to give the product as a pale yellow solid (6.2 g, 65% yield). LCMS: Calculated exact mass = 504.2; experimental value [M+H] + (ESI) = 505.0.

[0660] tert-Butyl 4-(4-amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)piperidine-1-carboxylate

[0661] A mixture of tert-butyl 4-(4-chloro-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)piperidine-1-carboxylate (500 mg, 0.99 mmol) in dioxane (2.5 mL) and NH 4 OH (2.5 mL) was reacted in a microwave reactor at 120 °C for 8 hours. After cooling to room temperature, the reaction mixture was concentrated to give the product as a pale yellow solid (450 mg, 93% yield). LCMS: Calculated exact mass = 485.2; experimental value [M+H] + (ESI) = 486.0.

[0662] 5-(4-Phenoxyphenyl)-7-(piperidin-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0663] To a solution of tert-butyl 4-(4-amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)piperidine-1-carboxylate (960 mg, 1.98 mmol) in DCM (8 mL) was added dropwise TFA (8 mL). The reaction mixture was stirred at room temperature for 2 h. The reaction mixture was concentrated and 2 M sodium hydroxide was added thereto to adjust the pH to 8 and the mixture was extracted with DCM (15 mL × 2). The organic layer was concentrated to give the product as a white solid (890 mg). LCMS: calculated exact mass = 385.3; experimental value [M+H] + (ESI) = 386.3; 1 H NMR (400 MHz, DMSO-d 6 ) δ 8.95 (d, J = 10.7 Hz, 1H), 8.68 (d, J = 9.8 Hz, 1H), 8.50 (s, 1H), 7.65 (s, 1H), 7.51 (d, J = 8.5 Hz, 2H), 7.41 - 7.47 (m, 2H), 7.07 - 7.22 (m, 5H), 4.92 - 5.06 (m, 2H), 3.48 (d, J = 12.5 Hz, 2H), 2.23 - 2.36 (m, 2H), 2.11 - 2.23 (m, 2H), 2.02 - 2.11 (m, 3H).

[0664] Example 147

[0665] 3-(2-Fluoro-4-phenoxyphenyl)-1-(1'-methyl-[1,4'-bipiperidin]-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine

[0666]

[0667] 3-(2-Fluoro-4-phenoxyphenyl)-1-(piperidin-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine

[0668] 3-Iodo-1-(piperidin-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (500 mg, 1.453 mmol), 2-(2-fluoro-4-phenoxyphenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (639 mg, 2.034 mmol), Pd(dppf)Cl 2 (212 mg, 0.291 mmol) and Na 2 CO 3 (461 mg, 4.358 mmol) in dioxane-H 2The mixture in O (8 mL - 1 mL) was heated overnight at 85 °C under an inert atmosphere. After cooling to room temperature, the reaction mixture was concentrated and purified by flash column chromatography (MeOH / DCM, 0 to 10% gradient) to afford the product as a semi-solid (300 mg, 51% yield).

[0669] 3-(2-Fluoro-4-phenoxyphenyl)-1-(1'-methyl-[1,4'-bipiperidin]-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine

[0670] To a mixture of 3-(2-fluoro-4-phenoxyphenyl)-1-(piperidin-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (150 mg, 0.37 mmol) and 1-methylpiperidin-4-one (126 mg, 1.11 mmol) in DCE (6 mL) was added 5 g of molecular sieve and 10 mL of AcOH. The mixture was stirred at 50 °C under N 2 atmosphere for 50 min. After cooling it to room temperature, NaBH(OAc) 3 (157 mg, 0.74 mmol) was added portionwise and the mixture was stirred overnight at room temperature. The reaction was monitored by TLC and LCMS until the starting materials were completely consumed. It was then filtered and concentrated, and Prep-TLC (15% MeOH / DCM) and Prep-HPLC (acetonitrile / water, 0.1% TFA) gave the desired product (10 mg, 5.4% yield). LCMS: calculated exact mass = 501.27; experimental value [M+H] + (ESI) = 502.41; 1 H NMR (DMSO-d 6 ) δ ppm: 10.01 (br.s., 1H), 8.29 - 8.35 (m, 1H), 7.44 - 7.58 (m, 3H), 7.25 (t, J = 7.4 Hz, 1H), 7.19 (d, J = 8.0 Hz, 2H), 7.04 (dd, J = 11.3, 2.1 Hz, 1H), 6.96 (dd, J = 8.4, 1.9 Hz, 1H), 5.13 (br.s., 1H), 4.39 (br.s., 3H), 3.59 - 3.67 (m, 4H), 3.34 (br.s., 2H), 3.02 (br.s., 2H), 2.79 (br.s., 3H), 2.17 - 2.39 (m, 4H), 1.94 (d, J = 10.0 Hz, 2H).

[0671] Example 148

[0672] 1-(4-(4-Amino-1-(1'-methyl-[1,4'-dipiperidin]-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-3-(5-(tert-butyl)isoxazol-3-yl)urea

[0673]

[0674] 3-Iodo-1-(1'-methyl-[1,4'-dipiperidin]-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine

[0675] A solution of 3-iodo-1-(piperidin-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (500 mg, 1.45 mmol) and 1-methylpiperidin-4-one (493.2 mg, 4.36 mmol) in tetrahydrofuran (60 mL) was stirred at room temperature for 1 hour. Subsequently, sodium triacetoxyborohydride (923.8 mg, 4.36 mmol) was added and stirring was continued for 16 hours. The mixture was concentrated and diluted with DCM-MeOH (250 mL, 10:1). The solution was filtered and concentrated to be purified by flash chromatography (DCM-MeOH-NH 3 (7.0 N in methanol): 100:10:1) to afford the product as a brown solid (400 mg, 62% yield). LCMS: calculated exact mass = 441.1; experimental value [M+H] + (ESI) = 441.7.

[0676] 1-(4-(4-Amino-1-(1'-methyl-[1,4'-dipiperidin]-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-3-(5-(tert-butyl)isoxazol-3-yl)urea

[0677] A solution of 3-iodo-1-(1'-methyl-[1,4'-dipiperidin]-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (200 mg, 0.45 mmol), (4-(3-(5-(tert-butyl)isoxazol-3-yl)ureido)phenyl)boronic acid (206 mg, 0.68 mmol), Pd(dppf)Cl 2 (66 mg, 0.09 mmol) and sodium carbonate (96 mg, 0.91 mmol) in 1,4-dioxane (8 mL) and water (0.8 mL) was reacted in a microwave reactor at 80 °C under a nitrogen atmosphere for 1 hour. After cooling to room temperature, the mixture was concentrated and purified by flash chromatography (DCM:MeOH:NH 3(7.0N in methanol) = 100:10:1) was purified to obtain a yellow solid (250 mg, 80% purity), which was further purified by Prep-HPLC to give the title compound as a white solid (105 mg, 40% yield). LCMS: calculated exact mass = 572.3; experimental value [M+H] + (ESI) = 572.8; 1 HNMR (400 MHz, DMSO-d 6 ) δ ppm: 9.71 (s, 2H), 9.17 (s, 1H), 8.30 (s, 1H), 7.66 (d, J = 8.5 Hz, 1H), 7.60 (d, J = 8.5 Hz, 2H), 6.52 (s, 1H), 5.11 - 5.25 (m, 1H), 3.35 - 3.72 (m, 9H), 3.02 (d, J = 8.9 Hz, 2H), 2.80 (s, 3H), 2.22 - 2.33 (m, 4H), 1.88 - 2.15 (m, 2H), 1.30 (s, 9H).

[0678] Using a similar procedure, the following compounds were prepared:

[0679]

[0680]

[0681]

[0682]

[0683]

[0684] Example 161

[0685] 2-(4-(4-Amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)piperidin-1-yl)propane-1,3-diol

[0686] 2-(4-(4-Amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)piperidin-1-yl)propane-1,3-diyl dibenzoate

[0687] A solution of 5-(4-phenoxyphenyl)-7-(piperidin-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (800 mg, 2.07 mmol) and 2-oxopropane-1,3-diyl dibenzoate (1.23 g, 4.15 mmol) in DCE (8 mL) was heated at 50 °C under N 2Stir for 2 h under the atmosphere. After cooling it to room temperature, add NaBH(OAc) portionwise 3 (1.3 g, 6.22 mmol), and stir the mixture overnight at room temperature under N 2 atmosphere. Monitor the reaction by TLC and LCMS until the starting materials are completely consumed. Then filter it, add saturated NaHCO 3 to the filtrate, and extract with DCM (30 mL × 3). Dry the organic layer over anhydrous Na 2 SO 4 . Concentrate to obtain the crude product. Purify the crude product by flash column chromatography (2 - 5% MeOH / DCM) to obtain the product as a white solid (150 mg, 10.8%). LCMS: calculated exact mass = 667.28; experimental value [M + H] + (ESI) = 667.71.

[0688] 2-(4-(4-Amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)piperidin-1-yl)propane-1,3-diol

[0689] Dissolve 2-(4-(4-Amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)piperidin-1-yl)propane-1,3-diyl dibenzoate (150 mg, 0.22 mmol) in a solution of 7.0 N NH 3 / MeOH (10 mL) and stir at 30 °C for 3 days. Monitor the reaction by TLC and LCMS until the starting materials are completely consumed. Then concentrate it and purify by flash column chromatography (5 - 10% MeOH / DCM) to obtain the product as a white solid (75 mg, 72.5% yield). LCMS: calculated exact mass = 459.23; experimental value [M + H] + (ESI) = 459.8; 1 H NMR (DMSO-d 6 ) δ ppm: 8.13 (s, 1H), 7.38 - 7.49 (m, 5H), 7.13 - 7.19 (m, 1H), 7.06 - 7.12 (m, 4H), 6.12 (br.s., 1H), 4.55 (t, J = 12.1 Hz, 1H), 4.29 (br.s., 2H), 3.43 - 3.57 (m, 4H), 2.97 (d, J = 10.7 Hz, 2H), 2.67 (t, J = 11.4 Hz, 2H), 2.55 - 2.62 (m, 1H), 1.98 - 2.06 (m, 2H), 1.87 (d, J = 10.7 Hz, 2H).

[0690] Example 162

[0691] 7-(1-(2-(Aminomethyl)-1,3-dioxolan-5-yl)piperidin-4-yl)-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0692]

[0693] 2-((5-(4-(4-Amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)piperidin-1-yl)-1,3-dioxolan-2-yl)methyl)isoindoline-1,3-dione

[0694] To a solution of 2-(4-(4-Amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)piperidin-1-yl)propane-1,3-diol (100 mg, 0.218 mmol) and 2-(1,3-dioxoisoindolin-2-yl)acetaldehyde (411 mg, 2.17 mmol) in chloroform (5 mL) was added anhydrous Na 2 SO 4 (927 mg, 6.53 mmol) and p-toluenesulfonic acid monohydrate (414 mg, 2.17 mmol). The mixture was refluxed overnight. It was then cooled to room temperature and saturated NaHCO 3 was added, and the mixture was extracted with DCM (25 mL × 3). The organic layer was concentrated to give a crude product. The crude product was purified by flash column chromatography (3 - 7% MeOH / DCM) to afford the product as a white solid (30 mg, 21.8% yield). LCMS: calculated exact mass = 510.31; experimental value [M + H] + (ESI) = 511.3.

[0695] 7-(1-(2-(Aminomethyl)-1,3-dioxolan-5-yl)piperidin-4-yl)-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0696] 2-((5-(4-(4-Amino-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)piperidin-1-yl)-1,3-dioxolan-2-yl)methyl)isoindoline-1,3-dione (30 mg, 0.048 mmol) was dissolved in 7.0 N NH 3The solution in / MeOH (20 mL) was stirred overnight at 30 °C. The reaction was monitored by TLC and LCMS until the starting material was completely consumed. It was then concentrated and purified by prep-TLC (MeOH:DCM = 40:3) to afford the product as a white solid (7 mg, 29% yield). LCMS: calculated exact mass = 500.25; experimental value [M+H] + (ESI) = 500.81; 1 H NMR (DMSO-d 6 ) δ ppm: 8.14 (s, 1H), 7.40 - 7.52 (m, 5H), 7.16 (t, J = 7.3 Hz, 1H), 7.10 (t, J = 6.9 Hz, 4H), 6.14 (br.s., 1H), 4.76 (d, J = 5.5 Hz, 1H), 4.59 (br.s., 1H), 4.13 (br.s., 1H), 3.70 - 3.81 (m, 1H), 3.58 (d, J = 6.7 Hz, 2H), 3.49 (d, J = 11.9 Hz, 1H), 3.17 (s, 3H), 3.04 (d, J = 10.4 Hz, 1H), 2.89 - 2.98 (m, 2H), 2.57 - 2.73 (m, 1H), 1.88 - 2.10 (m, 4H), 1.75 (s, 1H).

[0697] 4-(4-Amino-5-(2-fluoro-4-phenoxyphenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)cyclohexanone

[0698]

[0699] Ethyl 2-(1,4-dioxaspiro[4.5]dec-8-yl)-1H-imidazole-5-carboxylate

[0700] To a solution of NH 2 OH-HCl (447 g, 6.42 mol) in water (700 mL) was added portionwise NaHCO 3(787 g, 9.41 mol). Subsequently, 1,4-dioxaspiro[4.5]decane-8-carbonitrile (716 g, 4.28 mol) and EtOH (7 L) were added. The mixture was stirred at room temperature for 1 hour, and then heated to 80 °C for 22 hours. Subsequently, it was cooled to room temperature and ethyl propiolate (660 g, 6.73 mol) was added. The reaction mixture was heated to 80 °C for 7.5 hours. After cooling it to room temperature, the reaction mixture was filtered and the filtrate was concentrated and extracted with EtOAc. The organic layer was washed with brine and concentrated to obtain an orange solid (1.4 kg). Diphenyl ether (7 kg) was added and the reaction mixture was heated to 200 °C for 2 hours. The mixture was cooled to room temperature and PE (21 L) was added. After filtration, a black crude product (2 kg) was obtained, and the black crude product was purified by flash column chromatography (EtOAc 100%) to obtain the product as a yellow solid (850 g, 62% yield). 1 H NMR (400 MHz, CDCl 3 ) δ ppm: 7.63 (s, 1H), 4.34 (q, J = 7.1 Hz, 2H), 3.90 - 4.02 (m, 5H), 2.89 (br.s., 1H), 2.09 (dd, J = 13.2, 3.2 Hz, 2H), 1.86 (d, J = 12.3 Hz, 3H), 1.63 - 1.71 (m, 2H), 1.36 (t, J = 7.1 Hz, 3H).

[0701] 7-(1,4-Dioxaspiro[4.5]dec-8-yl)imidazo[5,1-f][1,2,4]triazin-4(3H)-one

[0702] To a solution of ethyl 2-(1,4-dioxaspiro[4.5]dec-8-yl)-1H-imidazole-5-carboxylate (312 g, 1.11 mol) in DMF (5 L) cooled in an ice bath was added dropwise LiHMDS (1.0 M, 1.11 L, 1.11 mol). The mixture was stirred for 2 hours, and then aminophenyldiphenylphosphinate (260 g, 1.11 mol) was added. The reaction mixture was stirred at room temperature overnight. DCM was added and the mixture was filtered. The filtrate was partially concentrated to obtain a DMF solution. Formamidine acetate (1.16 kg, 11.1 mol) was added and ethanol (5 L). Under N 2 atmosphere, the reaction mixture was heated to 85 °C for 2 days. The reaction mixture was heated to 100 °C for 3.5 hours. The mixture was concentrated to obtain 1 kg of a crude product, and the crude product was purified by flash column chromatography (MeOH / DCM, 0 to 10% gradient) to obtain a crude product as a light yellow wax (255 g), which was used without further purification. LC-MS: Calculated exact mass: 276.12; Experimental value [M + H] +(ESI) = 277.1。

[0703] 5-Bromo-7-(1,4-dioxaspiro[4.5]decan-8-yl)imidazo[5,1-f][1,2,4]triazin-4(3H)-one

[0704] To a solution of 7-(1,4-dioxaspiro[4.5]decan-8-yl)imidazo[5,1-f][1,2,4]triazin-4(3H)-one (100 g, 0.362 mol) in DMF (1.25 L) cooled in an ice bath was added portionwise NBS (77.3 g, 0.434 mol). The reaction mixture was stirred overnight at room temperature. The reaction was monitored by TLC and LCMS until the starting material was completely consumed. The reaction was quenched by adding 1 L of water. The solid was filtered and washed with water (200 mL × 3). The white solid was dried under reduced pressure to give a white powder product (98 g, 76.2% yield). LC-MS: Calculated exact mass: 354.03; Experimental value [M+H] + (ESI) = 355.13; 1 H NMR (400 MHz, DMSO-d 6 ) δ ppm: 11.85 (br.s., 1H), 7.90 (s, 1H), 3.83 - 3.92 (m, 4H), 3.17 (ddd, J = 10.8, 6.6, 4.3 Hz, 1H), 1.81 - 1.90 (m, 3H), 1.73 - 1.81 (m, 3H), 1.56 - 1.65 (m, 2H).

[0705] 5-Bromo-7-(1,4-dioxaspiro[4.5]decan-8-yl)imidazo[5,1-f][1,2,4]triazin-4-amine

[0706] To a suspension of 1H-1,2,4-triazole (97 g, 1.4 mol) in ACN (1.5 L) cooled in an ice bath was added dropwise POCl 3 (64 g, 0.42 mol), followed by addition of Et 3 N (170 g, 1.68 mol) and 5-bromo-7-(1,4-dioxaspiro[4.5]decan-8-yl)imidazo[5,1-f][1,2,4]triazin-4(3H)-one (50 g, 0.14 mol). The reaction mixture was stirred at room temperature for 6 hours. The solvent was removed under reduced pressure. Added NH 3(7.0 N, 1 L), heat the reactants to 80 °C for 1 hour, and then overnight at room temperature. Filter it and suspend the residue in water and aqueous NaOH solution. After filtration, a light yellow solid product (141 g, 82% yield) is obtained. LC-MS: calculated exact mass: 353.05; experimental value [M+H] + (ESI) = 354.2.

[0707] 5-(2-Fluoro-4-phenoxyphenyl)-7-(1,4-dioxaspiro[4.5]dec-8-yl)imidazo[5,1-f][1,2,4]triazin-4-amine

[0708] A mixture of 5-bromo-7-(1,4-dioxaspiro[4.5]dec-8-yl)imidazo[5,1-f][1,2,4]triazin-4-amine (40 g, 0.11 mol), 2-(2-fluoro-4-phenoxyphenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (48 g, 0.127 mol), Pd(dppf)Cl 2 (8 g, 0.011 mmol) 、 Na 2 CO 3 (35 g, 0.33 mol), and 700 mL of 1,4-dioxane (containing 10% water) is heated to 100 °C overnight. Cool the mixture to room temperature, filter and concentrate, extract the residue with DCM and H 2 O (1 L), dry and concentrate. Purify the crude product by flash column chromatography (MeOH / DCM, 0 to 2% gradient) to obtain a yellow solid product (42 g, 81% yield).

[0709] LC-MS: calculated exact mass: 461.19; experimental value [M+H] + (ESI) = 462.1.

[0710] 4-(4-Amino-5-(2-fluoro-4-phenoxyphenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)cyclohexanone

[0711] To a solution of 5-(2-fluoro-4-phenoxyphenyl)-7-(1,4-dioxaspiro[4.5]dec-8-yl)imidazo[5,1-f][1,2,4]triazin-4-amine (42 g, 0.091 mol) in acetone (400 mL) and THF (800 mL) was added 6N HCl (400 mL). The reaction mixture was stirred at 40 °C for 5 h. The mixture was cooled to room temperature and neutralized with aqueous NaOH. After concentration, the residue was extracted with DCM (300 mL × 3). The organic layer was washed with water, dried and concentrated. The crude product was purified by flash column chromatography to give the yellow solid product (14 g, 36.8% yield). LC-MS: Calculated exact mass: 417.16; Experimental value [M+H] + (ESI) = 418.31.

[0712] Example 163

[0713] 5-(2-Fluoro-4-phenoxyphenyl)-7-((trans)-4-(4-methylpiperazin-1-yl)cyclohexyl)imidazo[5,1-f][1,2,4]triazin-4-amine and

[0714] Example 164

[0715] 5-(2-Fluoro-4-phenoxyphenyl)-7-((cis)-4-(4-methylpiperazin-1-yl)cyclohexyl)imidazo[5,1-f][1,2,4]triazin-4-amine

[0716]

[0717] To a round bottom flask was added 4-(4-amino-5-(2-fluoro-4-phenoxyphenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)cyclohexanone (150 mg, 0.36 mmol), 1-methylpiperazine (180 mg, 1.8 mmol), DCM (30 mL) and molecular sieve. The mixture was stirred at room temperature for 15 min and NaBH(OAc) 3 (156 mg, 0.72 mol) was added portionwise. The reaction mixture was stirred at room temperature for 3 h. NaBH(AcO) 3 (156 mg, 0.72 mol) was added portionwise again. The reaction mixture was stirred at room temperature for 3.5 h. The reaction mixture was treated with saturated NaHCO 3Quench with aqueous solution. Then filter it and extract with DCM. Concentrate the organic layer and purify the crude product by flash column chromatography (MeOH / DCM, 0 to 10%) to obtain 5-(2-fluoro-4-phenoxyphenyl)-7-((trans)-4-(4-methylpiperazin-1-yl)cyclohexyl)imidazo[5,1-f][1,2,4]triazin-4-amine as a pale yellow solid (10 mg, 5.5% yield). LC-MS: Calculated exact mass: 501.27; Experimental value [M+H] + (ESI)=502.2; 1 H NMR(DMSO-d 6 )δ ppm: 7.89(s, 1H), 7.41 - 7.53(m, 3H), 7.15 - 7.25(m, 3H), 6.97(dd, J = 11.1, 2.4 Hz, 1H), 6.91(dd, J = 8.5, 2.4 Hz, 1H), 3.12 - 3.20(m, 1H), 2.63 - 2.78(m, 2H), 2.60(br.s., 2H), 2.53 - 2.57(m, 1H), 2.21 - 2.44(m, 5H), 1.94 - 2.16(m, 4H), 1.91(br.s., 2H), 1.59 - 1.74(m, 2H), 1.42(br.s., 2H), 1.34(d, J = 7.3 Hz, 1H); and 5-(2-fluoro-4-phenoxyphenyl)-7-((cis)-4-(4-methylpiperazin-1-yl)cyclohexyl)imidazo[5,1-f][1,2,4]triazin-4-amine (40 mg, 22% yield); 1 H NMR(400 MHz, DMSO)δ 9.79(s, 1H), 8.16(s, 1H), 7.89(s, 1H), 7.54 - 7.43(m, 3H), 7.23(t, J = 7.4 Hz, 1H), 7.20 - 7.15(m, 2H), 6.96(ddd, J = 10.8, 9.8, 2.3 Hz, 2H), 3.42(s, 1H), 2.99(s, 4H), 2.67(s, 3H), 2.27(d, J = 47.5 Hz, 4H), 2.13 - 1.93(m, 4H), 1.72(s, 2H), 1.59(s, 2H).

[0718] Example 165

[0719] 5-(2-fluoro-4-phenoxyphenyl)-7-((trans)-4-((S)-3-methylpiperazin-1-yl)cyclohexyl)imidazo[5,1-f][1,2,4]triazin-4-amine

[0720] and

[0721] Example 166

[0722] 5-(2-Fluoro-4-phenoxyphenyl)-7-((cis)-4-((S)-3-methylpiperazin-1-yl)cyclohexyl)imidazo[5,1-f][1,2,4]triazin-4-amine

[0723]

[0724] (S)-tert-Butyl 4-((trans)-4-(4-amino-5-(2-fluoro-4-phenoxyphenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)cyclohexyl)-2-methylpiperazine-1-carboxylate

[0725] and

[0726] (S)-tert-Butyl 4-((cis)-4-(4-amino-5-(2-fluoro-4-phenoxyphenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)cyclohexyl)-2-methylpiperazine-1-carboxylate

[0727] To a mixture of 4-(4-amino-5-(2-fluoro-4-phenoxyphenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)cyclohexanone (250 mg, 0.60 mmol) and (S)-tert-butyl 2-methylpiperazine-1-carboxylate (250 mg, 1.2 mmol) in DCM (30 mL) was added 4A molecular sieves. The mixture was stirred at room temperature for 2 hours, and then NaBH(OAc) 3 (31.67 g, 0.50 mol) was added portionwise. The reaction mixture was stirred at room temperature overnight. The reaction mixture was quenched with saturated NaHCO 3 aqueous solution. It was then filtered and extracted with DCM. The organic layer was collected and concentrated. The crude product was purified by flash column chromatography (MeOH / DCM, 0 to 5% gradient) to afford a mixture of two products as white solids (300 mg, 83% yield).

[0728] LC-MS: Calculated exact mass: 601.32; Experimental value [M+H] + (ESI) = 602.1.

[0729] 5-(2-Fluoro-4-phenoxyphenyl)-7-((trans)-4-((S)-3-methylpiperazin-1-yl)cyclohexyl)imidazo[5,1-f][1,2,4]triazin-4-amine

[0730] and

[0731] 5-(2-Fluoro-4-phenoxyphenyl)-7-((cis)-4-((S)-3-methylpiperazin-1-yl)cyclohexyl)imidazo[5,1-f][1,2,4]triazin-4-amine

[0732] To a solution of tert-butyl (S)-4-(4-(4-amino-5-(2-fluoro-4-phenoxyphenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)cyclohexyl)-2-methylpiperazine-1-carboxylate (300 mg, 0.50 mmol) in DCM (10 mL) was added TFA (2 mL). The mixture was stirred at room temperature for 3 h and then concentrated. Saturated NaHCO 3 aqueous solution and DCM were added and the mixture was extracted. The organic layer was collected and dried and concentrated. The crude product was purified by flash column chromatography (MeOH / DCM, 0 to 10% gradient) to afford 5-(2-fluoro-4-phenoxyphenyl)-7-((trans)-4-((S)-3-methylpiperazin-1-yl)cyclohexyl)imidazo[5,1-f][1,2,4]triazin-4-amine (25 mg, 10% yield). LC-MS: calculated exact mass: 501.27; experimental value [M+H] + (ESI)=502.1; 1 H NMR (400 MHz, DMSO-d 6 ) δ ppm: 8.31 (br.s., 1H), 7.89 (s, 1H), 7.42 - 7.52 (m, 3H), 7.15 - 7.26 (m, 3H), 6.98 (dd, J = 11.1, 2.4 Hz, 1H), 6.92 (dd, J = 8.5, 2.3 Hz, 1H), 3.18 (d, J = 12.5 Hz, 3H), 2.95 (br.s., 3H), 2.20 - 2.35 (m, 2H), 2.08 (d, J = 12.9 Hz, 2H), 1.89 (d, J = 9.6 Hz, 2H), 1.61 - 1.75 (m, 2H), 1.43 (d, J = 12.0 Hz, 2H), 1.19 (d, J = 6.5 Hz, 3H); and 5-(2-fluoro-4-phenoxyphenyl)-7-((cis)-4-((S)-3-methylpiperazin-1-yl)cyclohexyl)imidazo[5,1-f][1,2,4]triazin-4-amine (150 mg, 60% yield): 11H NMR (400 MHz, DMSO) δ 8.74 (s, 1H), 8.26 (s, 1H), 7.89 (s, 1H), 7.54 - 7.42 (m, 3H), 7.23 (t, J = 7.4 Hz, 1H), 7.20 - 7.15 (m, 2H), 6.96 (ddd, J = 10.9, 9.8, 2.4 Hz, 2H), 5.76 (s, 1H), 4.09 (s, 1H), 3.44 (s, 1H), 3.17 (s, 3H), 3.07 - 2.90 (m, 3H), 2.25 (s, 1H), 2.05 (d, J = 11.4 Hz, 2H), 1.92 (s, 2H), 1.72 (s, 2H), 1.59 (s, 2H), 1.18 (d, J = 6.5 Hz, 3H).

[0733] 4-(4-Amino-5-(4-phenoxyphenyl)pyrrolo[2,1-f][1,2,4]triazin-7-yl)cyclohexan-1-one

[0734]

[0735] 7-(1,4-Dioxaspiro[4.5]dec-7-en-8-yl)pyrrolo[2,1-f][1,2,4]triazin-4-amine

[0736] A mixture of 7-bromopyrrolo[2,1-f][1,2,4]triazin-4-amine (0.6 g, 2.82 mmol), 4,4,5,5-tetramethyl-2-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)-1,3,2-dioxaborolane (0.9 g, 3.38 mmol), Pd(dppf)Cl 2 (0.2 g, 0.28 mmol) and Na 2 CO 3 (0.9 g, 8.46 mmol) in 1,4-dioxane (50 mL) containing 10% water was stirred at 100 °C overnight. The mixture was cooled to room temperature, filtered and the filtrate was concentrated. The crude product was purified by flash column chromatography (PE:EtOAc = 1:1) to afford the product as a yellow solid (400 mg, 52% yield). LCMS: Calculated exact mass = 272.1; Experimental value [M+H] + (ESI) = 273.0.

[0737] 7-(1,4-Dioxaspiro[4.5]dec-8-yl)pyrrolo[2,1-f][1,2,4]triazin-4-amine

[0738] To a mixture of 7-(1,4-dioxaspiro[4.5]decan-7-en-8-yl)pyrrolo[2,1-f][1,2,4]triazin-4-amine (400 mg, 1.47 mmol) in MeOH / EtOAc (100 mL) was added Pd (80 mg, 10% on activated carbon). The mixture was stirred overnight at room temperature under H 2 atmosphere. Subsequently, it was filtered and the filtrate was concentrated to afford the product as a yellow solid (290 mg, 72% yield). LCMS: calculated exact mass = 274.1; experimental value [M+H] + (ESI) = 275.2.

[0739] 5-Bromo-7-(1,4-dioxaspiro[4.5]decan-8-yl)pyrrolo[2,1-f][1,2,4]triazin-4-amine

[0740] To a solution of 7-(1,4-dioxaspiro[4.5]decan-8-yl)pyrrolo[2,1-f][1,2,4]triazin-4-amine (290 mg, 1.06 mmol) in DCM (100 mL) was added NBS (226 mg, 1.27 mmol). The mixture was stirred at room temperature for 2.5 h. The mixture was quenched with saturated NaHCO 3 aqueous solution, extracted with DCM, dried and concentrated. The residue was purified by flash column chromatography (PE:EtOAc = 2:1) to afford the product as a pale yellow solid (250 mg, 67%). LCMS: calculated exact mass = 352.0; experimental value [M+H] + (ESI) = 353.1.

[0741] 5-(4-Phenoxyphenyl)-7-(1,4-dioxaspiro[4.5]decan-8-yl)pyrrolo[2,1-f][1,2,4]triazin-4-amine

[0742] 5-Bromo-7-(1,4-dioxaspiro[4.5]decan-8-yl)pyrrolo[2,1-f][1,2,4]triazin-4-amine (230 mg, 0.65 mmol), (4-phenoxyphenyl)boronic acid (418 mg, 1.95 mmol), Pd(dppf)Cl 2 (95 mg, 0.13 mmol) and Na 2 CO 3(276 mg, 2.6 mmol) in a mixture of 1,4-dioxane (20 mL) containing 10% water was stirred overnight at 100 °C. The mixture was cooled to room temperature, filtered and the filtrate was concentrated. The crude product was purified by flash column chromatography (PE:EtOAc = 1:1) to obtain the product as a yellow solid (125 mg, 42% yield). LCMS: calculated exact mass = 442.2; experimental value [M+H] + (ESI) = 443.1.

[0743] 4-(4-Amino-5-(4-phenoxyphenyl)pyrrolo[2,1-f][1,2,4]triazin-7-yl)cyclohexan-1-one

[0744] A mixture of 5-(4-phenoxyphenyl)-7-(1,4-dioxaspiro[4.5]decan-8-yl)pyrrolo[2,1-f][1,2,4]triazin-4-amine (120 mg, 0.27 mmol) in THF (2 mL), acetone (5 mL) and 6.0 N HCl (1 mL) was stirred at room temperature for 4 h. Subsequently, it was quenched with aqueous NaOH and extracted with DCM, dried and concentrated to obtain the crude product as a pale yellow solid (95 mg, 88% yield). LCMS: calculated exact mass = 398.2; experimental value [M+H] + (ESI) = 399.1.

[0745] 4-(4-Amino-5-bromopyrrolo[2,1-f][1,2,4]triazin-7-yl)cyclohexanone

[0746]

[0747] 7-(1,4-Dioxaspiro[4.5]decan-7-en-8-yl)pyrrolo[2,1-f][1,2,4]triazin-4-amine

[0748] 7-Bromopyrrolo[2,1-f][1,2,4]triazin-4-amine (1.6 g, 7.5 mmol), 4,4,5,5-tetramethyl-2-(1,4-dioxaspiro[4.5]decan-7-en-8-yl)-1,3,2-dioxaborolane (2.4 g, 9.0 mmol), Pd(dppf)Cl 2 (550 mg, 0.75 mmol) and Na 2 CO 3 (2.4 g, 22.5 mmol) in dioxane-H 2The mixture in O (50 mL - 5 mL) was heated overnight at 100 °C under an Ar atmosphere. After cooling to room temperature, the reaction mixture was concentrated and extracted with DCM (200 mL × 4). The organic layer was concentrated. The crude product was purified by flash column chromatography (PE:EA = 1:1) to give the product (900 mg, 36.7% yield). LCMS: Calculated exact mass = 272.1; experimental value [M+H] + (ESI) = 272.8.

[0749] 7-(1,4-Dioxaspiro[4.5]dec-8-yl)pyrrolo[2,1-f][1,2,4]triazin-4-amine

[0750] A mixture of 7-(1,4-Dioxaspiro[4.5]dec-7-en-8-yl)pyrrolo[2,1-f][1,2,4]triazin-4-amine (900 mg, 3.3 mmol) and Pd / C (180 mg) in MeOH-EtOAc (200 mL) was hydrogenated overnight under a balloon. The reaction mixture was filtered through Celite and washed with (DCM / MeOH = 10 / 1). The filtrate was concentrated to give the product as a pale yellow solid (900 mg, 99% yield). LCMS: Calculated exact mass = 274.1; experimental value [M+H] + (ESI) = 274.8.

[0751] 5-Bromo-7-(1,4-dioxaspiro[4.5]dec-8-yl)pyrrolo[2,1-f][1,2,4]triazin-4-amine

[0752] To a mixture of 7-(1,4-Dioxaspiro[4.5]dec-8-yl)pyrrolo[2,1-f][1,2,4]triazin-4-amine (900 mg, 3.3 mmol) in DCM (200 mL) was added NBS (700 mg, 3.9 mmol). The mixture was stirred at room temperature for 2 h. The reaction mixture was quenched with NaHCO 3 and extracted with DCM, dried and concentrated to give the crude product (1.15 g), which was used without further purification. LCMS: Calculated exact mass = 452.1; experimental value [M+H] + (ESI) = 452.8.

[0753] 4-(4-Amino-5-bromopyrrolo[2,1-f][1,2,4]triazin-7-yl)cyclohexanone

[0754] To a suspension of 5-bromo-7-(1,4-dioxaspiro[4.5]dec-8-yl)pyrrolo[2,1-f][1,2,4]triazin-4-amine (1.1 g, 3.1 mmol) in THF (25 mL) and acetone (25 mL) was added 6N HCl (10 mL). The reaction mixture was heated at 50 °C for 2 h. It was neutralized with an aqueous NaHCO 3 solution, extracted with DCM, dried and concentrated. The residue was purified by flash column chromatography (PE:EA = 1:1) to give the product (600 mg, 62.6% yield). LCMS: calculated exact mass = 308.0; experimental value [M+H] + (ESI) = 308.8.

[0755]

[0756] 5-bromo-7-((trans)-4-(4-methylpiperazin-1-yl)cyclohexyl)pyrrolo[2,1-f][1,2,4]triazin-4-amine

[0757] and

[0758] 5-bromo-7-((cis)-4-(4-methylpiperazin-1-yl)cyclohexyl)pyrrolo[2,1-f][1,2,4]triazin-4-amine

[0759] A mixture of 4-(4-amino-5-bromopyrrolo[2,1-f][1,2,4]triazin-7-yl)cyclohexanone (200 mg, 0.67 mmol), 1-methylpiperazine (134 mg, 1.34 mmol), and titanium(IV) isopropoxide (758 mg, 2.67 mmol) in DCM (30 mL) was stirred at room temperature. After stirring for 2 h, sodium triacetoxyborohydride (560 mg, 2.67 mmol) was added. It was stirred at room temperature for 16 h, then diluted with DCM. Subsequently, an aqueous NaHCO 3 solution was added. The mixture was filtered. The organic layer was dried and concentrated in vacuo. The residue was purified by flash column chromatography (DCM:MeOH = 10:1) to give 5-bromo-7-((cis)-4-(4-methylpiperazin-1-yl)cyclohexyl)pyrrolo[2,1-f][1,2,4]triazin-4-amine as a white solid (80 mg, 30% yield) and 5-bromo-7-((trans)-4-(4-methylpiperazin-1-yl)cyclohexyl)pyrrolo[2,1-f][1,2,4]triazin-4-amine as a pale yellow solid (60 mg, 22% yield).

[0760] 4-(4-amino-5-(4-phenoxyphenyl)pyrrolo[2,1-f][1,2,4]triazin-7-yl)cyclohexanone

[0761]

[0762] 4-(4-Amino-5-(4-phenoxyphenyl)pyrrolo[2,1-f][1,2,4]triazin-7-yl)cyclohexanone

[0763] A mixture of 4-(4-amino-5-bromopyrrolo[2,1-f][1,2,4]triazin-7-yl)cyclohexanone (60 mg, 0.19 mmol), (4-phenoxyphenyl)boronic acid (82 g, 0.38 mmol), Pd(dppf)Cl 2 (14 mg, 0.019 mmol), and Na 2 CO 3 (41 mg, 0.38 mmol) in dioxane-H 2 O (10 mL - 1 mL) was heated at 85 °C for 2 h under an Ar atmosphere. After cooling to room temperature, the reaction mixture was concentrated and extracted with DCM (50 mL × 4). The organic layer was concentrated and purified by flash column chromatography (PE:EA = 6:1) to give the product as a pale yellow solid (60 mg, 79% yield). LCMS: Calculated exact mass = 398.1; experimental value [M+H] + (ESI) = 398.9.

[0764] Example 167

[0765] 7-((cis)-4-(4-Methylpiperazin-1-yl)cyclohexyl)-5-(4-phenoxyphenyl)pyrrolo[2,1-f][1,2,4]triazin-4-amine and

[0766] Example 168

[0767] 7-((trans)-4-(4-Methylpiperazin-1-yl)cyclohexyl)-5-(4-phenoxyphenyl)pyrrolo[2,1-f][1,2,4]triazin-4-amine

[0768]

[0769] A mixture of 4-(4-amino-5-(4-phenoxyphenyl)pyrrolo[2,1-f][1,2,4]triazin-7-yl)cyclohex-1-one (90 mg, 0.22 mmol) and 1-methylpiperazine (110 mg, 1.1 mmol) in 1,2-dichloroethane (5 mL) containing AcOH (2 drops) was stirred at room temperature under an N 2 atmosphere for 1 h. Then NaBH(OAc) 3 (370 mg, 1.76 mmol) was added portionwise. At room temperature under an N 2Stir the mixture for 3 hours under the atmosphere. Monitor the reaction by TLC and LCMS until the starting materials are completely depleted. Then filter and concentrate it, and purify the crude product by flash (1 - 10% MeOH / DCM) to obtain 7-((cis)-4-(4-methylpiperazin-1-yl)cyclohexyl)-5-(4-phenoxyphenyl)pyrrolo[2,1-f][1,2,4]triazin-4-amine as a colorless solid (28 mg, 26% yield). LCMS: calculated exact mass = 442.2; experimental value [M+H] + (ESI) = 443.1; 1 H NMR (DMSO-d6) δ 7.89 (s, 1H), 7.38 - 7.51 (m, 4H), 7.17 (t, J = 7.3 Hz, 1H), 7.10 (dd, J = 7.9, 3.7 Hz, 4H), 6.54 (br.s., 1H), 2.67 (br.s., 3H), 2.32 (br.s., 5H), 1.88 (br.s., 5H), 1.75 (br.s., 3H), 1.58 (br.s., 2H); and 7-((trans)-4-(4-methylpiperazin-1-yl)cyclohexyl)-5-(4-phenoxyphenyl)pyrrolo[2,1-f][1,2,4]triazin-4-amine as a colorless solid (25 mg, 23% yield). LCMS: calculated exact mass = 442.2; experimental value [M+H] + (ESI) = 443.1; 1 H NMR (DMSO-d6) δ 7.89 (s, 1H), 7.39 - 7.47 (m, 4H), 7.17 (t, J = 7.3 Hz, 1H), 7.09 (t, J = 8.4 Hz, 4H), 6.50 (s, 1H), 3.04 (d, J = 7.9 Hz, 1H), 2.58 (br.s., 3H), 2.33 (br.s., 4H), 2.18 - 2.28 (m, 4H), 2.13 (d, J = 11.6 Hz, 3H), 1.92 (br.s., 2H), 1.37 - 1.56 (m, 4H).

[0770] 7-((trans)-4-((S)-3-methylpiperazin-1-yl)cyclohexyl)-5-(4-phenoxyphenyl)pyrrolo[2,1-f][1,2,4]triazin-4-amine

[0771]

[0772] (S)-tert-butyl 4-((trans)-4-(4-amino-5-(4-phenoxyphenyl)pyrrolo[2,1-f][1,2,4]triazin-7-yl)cyclohexyl)-2-methylpiperazine-1-carboxylate

[0773] 4-(4-Amino-5-(4-phenoxyphenyl)pyrrolo[2,1-f][1,2,4]triazin-7-yl)cyclohexanone (50 mg, 0.12 mmol), (S)-tert-butyl 2-methylpiperazine-1-carboxylate (50 mg, 0.24 mmol), and Ti(OiPr) 4 (142 mg, 0.48 mmol) in DCM (10 mL) was stirred at room temperature for 2 h. NaBH(OAc) 3 (100 mg, 0.48 mol) was added. It was stirred overnight at room temperature. Quenched with MeOH. Subsequently, aqueous NaHCO 3 solution and DCM were added, and it was filtered. The filtrate was extracted with DCM. The organic layer was collected and concentrated. The residue was purified by flash column chromatography (DCM:MeOH = 10:1) to give (S)-tert-butyl 4-((trans)-4-(4-amino-5-(4-phenoxyphenyl)pyrrolo[2,1-f][1,2,4]triazin-7-yl)cyclohexyl)-2-methylpiperazine-1-carboxylate (25 mg, 36% yield). LCMS: calculated exact mass = 582.3; experimental value [M+H] + (ESI) = 583.1; and (S)-tert-butyl 4-((cis)-4-(4-amino-5-(4-phenoxyphenyl)pyrrolo[2,1-f][1,2,4]triazin-7-yl)cyclohexyl)-2-methylpiperazine-1-carboxylate (21 mg, 30% yield) as a white solid. LCMS: calculated exact mass = 582.3; experimental value [M+H] + (ESI) = 583.1.

[0774] Example 169

[0775] 7-((trans)-4-((S)-3-Methylpiperazin-1-yl)cyclohexyl)-5-(4-phenoxyphenyl)pyrrolo[2,1-f][1,2,4]triazin-4-amine

[0776]

[0777] 7-((trans)-4-((S)-3-Methylpiperazin-1-yl)cyclohexyl)-5-(4-phenoxyphenyl)pyrrolo[2,1-f][1,2,4]triazin-4-amine

[0778] To a solution of tert-butyl (S)-4-((trans)-4-(4-amino-5-(4-phenoxyphenyl)pyrrolo[2,1-f][1,2,4]triazin-7-yl)cyclohexyl)-2-methylpiperazine-1-carboxylate (25 mg, 0.04 mmol) in DCM (10 mL) was added dropwise TFA (0.5 mL). The reaction mixture was stirred at room temperature for 3 h, then concentrated and the residue was purified by Prep-HPLC to give the product as a white solid (18 mg, 93% yield). LCMS: calculated exact mass = 482.3; found [M+H] + (ESI) = 483.1; 1 H NMR (400 MHz, methanol-d 4 ) δ ppm 7.94 (s, 1H) 7.41 (d, J = 8.54 Hz, 2H) 7.31 (t, J = 7.93 Hz, 2H) 7.08 (t, J = 7.32 Hz, 1H) 7.00 (d, J = 7.93 Hz, 2H) 7.04 (d, J = 8.55 Hz, 2H) 6.68 (s, 1H) 3.55 - 3.68 (m, 4H) 3.29 - 3.41 (m, 1H) 2.99 (t, J = 12.05 Hz, 1H) 2.12 - 2.29 (m, 4H) 1.58 - 1.76 (m, 4H) 1.30 - 1.36 (m, 3H).

[0779] Example 170

[0780] 7-((cis)-4-((S)-3-methylpiperazin-1-yl)cyclohexyl)-5-(4-phenoxyphenyl)pyrrolo[2,1-f][1,2,4]triazin-4-amine

[0781]

[0782] 7-((cis)-4-((S)-3-methylpiperazin-1-yl)cyclohexyl)-5-(4-phenoxyphenyl)pyrrolo[2,1-f][1,2,4]triazin-4-amine

[0783] To a solution of tert-butyl (S)-4-((cis)-4-(4-amino-5-(4-phenoxyphenyl)pyrrolo[2,1-f][1,2,4]triazin-7-yl)cyclohexyl)-2-methylpiperazine-1-carboxylate (21 mg, 0.04 mmol) in DCM (10 mL) was added dropwise TFA (0.5 mL). The reaction mixture was stirred at room temperature for 4 h. The mixture was evaporated and dissolved in DCM, and washed with NaHCO 3Solution and water washings. The organic layer was dried and concentrated in vacuo to afford the title compound as a pale yellow solid (16 mg, 82% yield). LC-MS: calculated exact mass: 482.3; found [M+H] + (ESI) = 483.7; 1 HNMR (400 MHz, MeOD) δ 8.03 (s, 1H), 7.54 (d, J = 8.7 Hz, 2H), 7.46 - 7.40 (m, 2H), 7.20 (t, J = 7.4 Hz, 1H), 7.13 (dd, J = 15.3, 8.1 Hz, 4H), 6.86 (s, 1H), 3.52 (d, J = 14.0 Hz, 6H), 2.83 (s, 1H), 2.74 - 2.62 (m, 1H), 2.48 (s, 1H), 2.18 (s, 2H), 1.95 (d, J = 19.6 Hz, 7H), 1.37 (d, J = 6.5 Hz, 3H).

[0784] Example 171

[0785] 1-(4-(4-Amino-7-((trans)-4-(4-methylpiperazin-1-yl)cyclohexyl)pyrrolo[2,1-f][1,2,4]triazin-5-yl)phenyl)-3-(5-(tert-butyl)furan-3-yl)urea

[0786] and

[0787] Example 172

[0788] 1-(4-(4-Amino-7-((cis)-4-(4-methylpiperazin-1-yl)cyclohexyl)pyrrolo[2,1-f][1,2,4]triazin-5-yl)phenyl)-3-(5-(tert-butyl)furan-3-yl)urea

[0789]

[0790] A mixture of 5-bromo-7-((trans)-4-(4-methylpiperazin-1-yl)cyclohexyl)pyrrolo[2,1-f][1,2,4]triazin-4-amine and 5-bromo-7-((cis)-4-(4-methylpiperazin-1-yl)cyclohexyl)pyrrolo[2,1-f][1,2,4]triazin-4-amine (80 mg, 0.2 mmol), (4-(3-(5-(tert-butyl)furan-3-yl)ureido)phenyl)boronic acid (74 mg, 0.24 mmol), sodium carbonate (64 mg, 0.6 mmol) and Pd(dppf)Cl 2 (30 mg, 0.04 mmol) in dioxane - H 2A mixture of O (10 mL - 1 mL) was heated at 100 °C for 3 hours under an inert atmosphere. The reactants were concentrated in vacuo. The residue was dissolved in DCM, washed with water, dried, evaporated and purified by Prep-TLC (DCM:MeOH = 10:1) to give 1-(4-(4-amino-7-((trans)-4-(4-methylpiperazin-1-yl)cyclohexyl)pyrrolo[2,1-f][1,2,4]triazin-5-yl)phenyl)-3-(5-(tert-butyl)furan-3-yl)urea as a white solid (5 mg, 4% yield). LC-MS: Calculated exact mass: 571.3; Experimental value [M+H] + (ESI) = 572.3; 1 H NMR (400 MHz, MeOD) δ 7.82 (s, 1H), 7.61 (d, J = 8.5 Hz, 2H), 7.45 (d, J = 8.5 Hz, 2H), 6.52 (s, 1H), 6.42 (s, 1H), 3.20 (s, 2H), 2.78 (d, J = 65.6 Hz, 10H), 2.42 (s, 3H), 2.28 (d, J = 10.8 Hz, 3H), 2.14 (d, J = 10.2 Hz, 2H), 1.68 - 1.50 (m, 5H), 1.38 (s, 10H); and 1-(4-(4-amino-7-((cis)-4-(4-methylpiperazin-1-yl)cyclohexyl)pyrrolo[2,1-f][1,2,4]triazin-5-yl)phenyl)-3-(5-(tert-butyl)furan-3-yl)urea as a white solid (21 mg, 18% yield). LC-MS: Calculated exact mass: 571.3; Experimental value [M+H] + (ESI) = 572.3; 1 H NMR (400 MHz, MeOD) δ 7.70 (s, 1H), 7.50 (d, J = 8.5 Hz, 2H), 7.34 (d, J = 8.5 Hz, 2H), 6.52 (s, 1H), 6.30 (s, 1H), 3.37 (s, 1H), 2.72 (s, 8H), 2.40 (s, 4H), 2.07 (s, 2H), 1.82 - 1.67 (m, 6H), 1.26 (s, 9H).

[0791] 8-Chloro-3-((trans)-4-(4-methylpiperazin-1-yl)cyclohexyl)imidazo[1,5-a]pyrazine

[0792] and

[0793] 8-Chloro-3-((cis)-4-(4-methylpiperazin-1-yl)cyclohexyl)imidazo[1,5-a]pyrazine

[0794]

[0795] N-((3-chloropyrazin-2-yl)methyl)-4-oxocyclohexanecarboxamide

[0796] To a solution of 4-oxocyclohexanecarboxylic acid (0.7 g, 3.9 mmol) and TEA (1.1 mL, 7.8 mmol) in DCM (10 mL) was added HATU (1.77 g, 4.67 mmol) and (3-chloropyrazin-2-yl)methylamine (1.77 g, 4.67 mmol). The reaction mixture was stirred at room temperature for 18 h, then quenched with water (20 mL) and extracted with DCM (20 mL×3). The organic layer was collected, washed with brine (20 mL) and dried over Na 2 SO 4 After concentration, the crude product was purified by flash column chromatography (EA 100%) to give the desired product as a yellow solid (680 mg, 65% yield). LCMS: calculated exact mass = 267.08; experimental value [M+H] + (ESI) = 268.0; 1 H NMR (400 MHz, DMSO-d 6 ) δ ppm: 8.63 (d, J = 2.42 Hz, 1H), 8.50 (t, J = 5.24 Hz, 1H), 8.44 (d, J = 2.42 Hz, 1H), 4.53 (d, J = 5.37 Hz, 2H), 2.72 (tt, J = 3.59, 10.64 Hz, 1H), 2.34 - 2.45 (m, 2H), 2.23 - 2.33 (m, 2H), 2.05 (dd, J = 3.76, 13.16 Hz, 2H), 1.73 - 1.88 (m, 2H).

[0797] N-((3-chloropyrazin-2-yl)methyl)-4-(4-methylpiperazin-1-yl)cyclohexanecarboxamide

[0798] At 0 °C under N 2 atmosphere, to a solution of N-((3-chloropyrazin-2-yl)methyl)-4-oxocyclohexanecarboxamide (2.2 g, 8.1 mmol), 1-methylpiperazine (1.2 g, 10.6 mmol) in DCM (30 mL) was added NaBH 3 CN (1023 mg, 16.2 mmol) and AcOH (292 mg, 4.87 mmol). The reaction mixture was stirred at 20 °C for 3 h. The reaction was quenched with NaHCO 3 solution (20 mL) and extracted with DCM (20 mL×3). The organic layer was washed with brine (20 mL) and dried over Na 2 SO 4Drying. After concentration, the crude product was purified by flash column chromatography (DCM:MeOH = 100:1 to 5:1) to give the desired product as a yellow solid (1.4 g, 60% yield). LCMS: Calculated exact mass = 351.18; Experimental value [M+H] + (ESI) = 352.0.

[0799] 8-Chloro-3-(4-(4-methylpiperazin-1-yl)cyclohexyl)imidazo[1,5-a]pyrazine

[0800] At 0 °C under N 2 atmosphere, DMI (1.37 g, 12 mmol) and POCl 3 (2.5 g, 16 mmol) were added dropwise to a solution of N-((3-chloropyrazin-2-yl)methyl)-4-(4-methylpiperazin-1-yl)cyclohexanecarboxamide (1.4 g, 4 mmol) in ACN (30 mL). The reaction mixture was stirred at reflux for 3 h. It was carefully quenched with NH 3 .H 2 O (20 mL) and ice and extracted with DCM (20 mL × 3). The organic layer was washed with brine (20 mL) and dried over Na 2 SO 4 4. After concentration, the crude product was purified by flash column chromatography (DCM:MeOH = 100:1 - 10:1) to give 8-chloro-3-((trans)-4-(4-methylpiperazin-1-yl)cyclohexyl)imidazo[1,5-a]pyrazine (240 mg, 18% yield). LCMS: Calculated exact mass = 333.17; Experimental value [M+H] + (ESI) = 334.1 and 8-chloro-3-((cis)-4-(4-methylpiperazin-1-yl)cyclohexyl)imidazo[1,5-a]pyrazine (340 mg, 25% yield). LCMS: Calculated exact mass = 333.17; Experimental value [M+H] + (ESI) = 334.1.

[0801] Example 173

[0802] 3-((trans)-4-(4-Methylpiperazin-1-yl)cyclohexyl)-1-(4-phenoxyphenyl)imidazo[1,5-a]pyrazin-8-amine

[0803]

[0804] 1-Bromo-8-chloro-3-((trans)-4-(4-methylpiperazin-1-yl)cyclohexyl)imidazo[1,5-a]pyrazine

[0805] At 20 °C under N 2 To a solution of 8-chloro-3-((trans)-4-(4-methylpiperazin-1-yl)cyclohexyl)imidazo[1,5-a]pyrazine (240 mg, 0.72 mmol) in DCM (10 mL) was added NBS (192 mg, 1.08 mmol) under N atmosphere. The reaction mixture was then stirred at reflux for 3 h. By LC-MS and TLC detection, the starting material was exhausted. The reaction mixture was quenched with NaHCO 3 (20 mL) and extracted with DCM (20 mL × 3). The organic layer was washed with brine (20 mL), dried over Na 2 SO 4 and concentrated in vacuo to give the desired product as a yellow solid (300 mg, 98% yield), which was used in the next step without further purification. LCMS: calculated exact mass = 411.08; found [M+H] + (ESI) = 412.1.

[0806] 1-Bromo-3-((trans)-4-(4-methylpiperazin-1-yl)cyclohexyl)imidazo[1,5-a]pyrazin-8-amine

[0807] To a solution of 1-bromo-8-chloro-3-((trans)-4-(4-methylpiperazin-1-yl)cyclohexyl)imidazo[1,5-a]pyrazine (300 mg, 0.70 mmol) in 1,4-dioxane (10 mL) in a sealed tube was added NH 3 .H 2 O (10 mL). The reaction mixture was then stirred at 120 °C for 48 h. By LC-MS and TLC detection, the starting material was nearly exhausted. It was extracted with EA (30 mL × 3). The organic layer was washed with brine (40 mL), dried over Na 2 SO 4 and concentrated in vacuo and purified by silica gel chromatography (DCM:MeOH = 100:1 - 10:1) to give the desired product as a yellow solid (100 mg, 34% yield).

[0808] 3-((trans)-4-(4-methylpiperazin-1-yl)cyclohexyl)-1-(4-phenoxyphenyl)imidazo[1,5-a]pyrazin-8-amine

[0809] 1-Bromo-3-((trans)-4-(4-methylpiperazin-1-yl)cyclohexyl)imidazo[1,5-a]pyrazin-8-amine (100 mg, 0.25 mmol) and 4-phenoxyphenylboronic acid (65 mg, 0.30 mmol), Na 2 CO 3(81 mg, 0.76 mmol) and Pd(dppf)Cl 2 (37 mg, 0.05 mmol) in a suspension of THF (15 mL) and water (3 mL) was stirred at 80 °C under N 2 atmosphere for 3 h. Detection by LC-MS and TLC showed that the starting materials were exhausted. The reaction mixture was concentrated in vacuo and purified by flash column chromatography (PE:EA = 10:1 - 3:1) and Prep-HPLC to give the desired product as a white solid (26 mg, 22% yield). LCMS: calculated exact mass = 482.28; experimental value [M+H] + (ESI) = 483.3; 1 H NMR (600 MHz, methanol-d 4 ) δ ppm: 7.74 (d, J = 5.72 Hz, 1H), 7.62 (d, J = 8.47 Hz, 2H), 7.41 (t, J = 7.78 Hz, 2H), 7.19 (t, J = 7.32 Hz, 1H), 7.15 (d, J = 8.47 Hz, 2H), 7.10 (d, J = 8.24 Hz, 2H), 6.99 (d, J = 5.49 Hz, 1H), 3.35 (s, 1H), 3.32 (br.s., 4H), 3.18 (t, J = 11.79 Hz, 1H), 2.71 (br.s., 3H), 2.15 (d, J = 9.84 Hz, 4H), 1.85 (q, J = 12.05 Hz, 2H), 1.58 - 1.70 (m, 2H), 1.26 - 1.37 (m, 2H).

[0810] Example 174

[0811] 3-((cis)-4-(4-Methylpiperazin-1-yl)cyclohexyl)-1-(4-phenoxyphenyl)imidazo[1,5-a]pyrazin-8-amine

[0812]

[0813] 1-Bromo-8-chloro-3-((cis)-4-(4-methylpiperazin-1-yl)cyclohexyl)imidazo[1,5-a]pyrazine

[0814] At 20 °C under N 2 atmosphere, NBS (128 mg, 0.72 mmol) was added to a solution of 8-chloro-3-((cis)-4-(4-methylpiperazin-1-yl)cyclohexyl)imidazo[1,5-a]pyrazine (200 mg, 0.6 mmol) in DCM (10 mL). Subsequently, the reaction mixture was stirred under reflux for 3 h. Detection by LC-MS and TLC showed that the starting materials were exhausted. The reaction mixture was passed through NaHCO3 The solution (20 mL) was quenched, extracted with DCM (20 mL × 3), washed with brine (20 mL), and dried over Na 2 SO 4 and concentrated in vacuo to give the desired product as a yellow solid (240 mg, 98%, yield), which was used in the next step without further purification. LCMS: Calculated exact mass = 411.08; experimental value [M+H] + (ESI) = 412.1.

[0815] 1-Bromo-3-((cis)-4-(4-methylpiperazin-1-yl)cyclohexyl)imidazo[1,5-a]pyrazin-8-amine

[0816] To a solution of 1-bromo-8-chloro-3-((cis)-4-(4-methylpiperazin-1-yl)cyclohexyl)imidazo[1,5-a]pyrazine (190 mg, 0.46 mmol) in 1,4-dioxane (10 mL) in a sealed tube was added NH 3 .H 2 O (10 mL). The reaction mixture was then stirred at 120 °C for 48 h. By LC-MS and TLC monitoring, the starting material was nearly exhausted. It was extracted with EA (30 mL × 3). The organic layer was washed with brine (40 mL), dried over Na 2 SO 4 and concentrated in vacuo. The crude product was purified by flash column chromatography (DCM:MeOH = 100:1 - 10:1) to give the desired product as a yellow solid (160 mg, 80% yield).

[0817] 3-((cis)-4-(4-methylpiperazin-1-yl)cyclohexyl)-1-(4-phenoxyphenyl)imidazo[1,5-a]pyrazin-8-amine

[0818] A suspension of 1-bromo-3-((cis)-4-(4-methylpiperazin-1-yl)cyclohexyl)imidazo[1,5-a]pyrazin-8-amine (160 mg, 0.4 mmol), 4-phenoxyphenylboronic acid (100 mg, 0.48 mmol), Na 2 CO 3 (128 mg, 1.2 mmol), and Pd(dppf)Cl 2 (60 mg, 0.08 mmol) in THF (15 mL) and water (3 mL) was heated at 80 °C under N 2Stir for 3 hours under the atmosphere. Detection by LC-MS and TLC showed that the starting materials were exhausted. The reaction mixture was concentrated in vacuo and purified by flash column chromatography eluting with (PE:EA = 10:1 - 3:1) and Prep-HPLC to afford the desired product as a white solid (40 mg, 40% yield). LCMS: calculated exact mass = 482.28; experimental value [M+H] + (ESI)=483.3; 1 H NMR (600 MHz, methanol-d 4 ) δ ppm: 7.76 (d, J = 5.95 Hz, 1H), 7.69 (d, J = 8.47 Hz, 2H), 7.42 (t, J = 7.90 Hz, 2H), 7.20 (t, J = 7.32 Hz, 1H), 7.15 (d, J = 8.47 Hz, 2H), 7.11 (d, J = 8.01 Hz, 2H), 6.97 (d, J = 5.95 Hz, 1H), 3.31 - 3.60 (m, 9H), 3.21 (br.s., 1H), 2.90 (s, 3H), 2.17 - 2.29 (m, 4H), 1.88 - 2.06 (m, 4H).

[0819] Example 175

[0820] 1-(4-(8-Amino-3-((trans)-4-(4-methylpiperazin-1-yl)cyclohexyl)imidazo[1,5-a]pyrazin-1-yl)phenyl)-3-(5-(tert-butyl)isoxazol-3-yl)urea

[0821]

[0822] A mixture of 5-bromo-7-((trans)-4-(4-methylpiperazin-1-yl)cyclohexyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (60 mg, 0.153 mmol), (4-(3-(5-(tert-butyl)isoxazol-3-yl)ureido)phenyl)boronic acid (70 mg, 0.23 mmol), Pd(dppf)Cl 2 (11 mg, 0.015 mmol) and Na 2 CO 3 (49 mg, 0.459 mmol) in dioxane-H 2 O (10:1, 22 mL) was heated to 80 °C. The reaction mixture was stirred for 2 hours. TLC (MeOH:DCM = 1:10) showed complete exhaustion of the starting materials. The solvent was evaporated in vacuo. The residue was extracted with DCM (100 mL). The combined organic layers were washed with 1 M NaOH (200 mL), dried over anhydrous Na 2 SO 4Dry and concentrate in vacuo. Purify the crude product by Prep-HPLC to give the product as a white solid (16 mg, 18% yield). LCMS: calculated exact mass = 574.3, experimental value [M+H] + (ESI) = 575.3; 1 HNMR(DMSO-d 6 ) δ ppm: 9.82 (s, 1H), 9.36 (s, 1H), 7.95 (s, 1H), 7.56 - 7.67 (m, 4H), 7.10 - 7.12 (m, 1H), 6.52 (s, 1H), 3.05 - 3.25 (m, 5H), 1.50 - 2.25 (m, 8H), 1.30 (s, 12H).

[0823] Example 176

[0824] 7 - ((cis)-4-(4-Methylpiperazin-1-yl)cyclohexyl)-5-(4-phenoxyphenyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine

[0825] and

[0826] Example 177

[0827] 7 - ((trans)-4-(4-Methylpiperazin-1-yl)cyclohexyl)-5-(4-phenoxyphenyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine

[0828]

[0829] 4-Methoxy-5-(4-phenoxyphenyl)-5H-pyrrolo[3,2-d]pyrimidine

[0830] A mixture of 4-methoxy-5H-pyrrolo[3,2-d]pyrimidine (650 mg, 4.36 mmol), 1-bromo-4-phenoxybenzene (1.3 g, 5.22 mmol), CuI (1.66 g, 8.72 mmol), (1R,2R)-N1,N2-dimethylcyclohexane-1,2-diamine (1.24 g, 8.72 mmol), K 2 CO 3 (1.8 g, 13.08 mmol) in DMSO (50 mL) was heated at 140 °C for 3 h under an Ar atmosphere. Cool to room temperature and filter, quench the filtrate with water and extract with EtOAc, dry and concentrate. Purify the residue by column chromatography (PE:EA = 3:1) to obtain the product (750 mg, 53% yield). LCMS: calculated exact mass = 317.1; experimental value [M+H] + (ESI) = 318.0.

[0831] 7-Bromo-4-methoxy-5-(4-phenoxyphenyl)-5H-pyrrolo[3,2-d]pyrimidine

[0832] To a mixture of 4-methoxy-5-(4-phenoxyphenyl)-5H-pyrrolo[3,2-d]pyrimidine (1 g, 3.2 mmol) in DCM (50 mL) was added NBS (670 mg, 3.8 mmol). The mixture was stirred at room temperature for 2 h. Quenched with saturated NaHCO 3 , extracted with DCM, dried and concentrated, and the residue was purified by column chromatography (PE:EA = 2:1) to afford the product as a yellow solid (1 g, 79% yield). LCMS: calculated exact mass = 395.0; experimental value [M+H] + (ESI) = 395.9.

[0833] 7-Bromo-5-(4-phenoxyphenyl)-5H-pyrrolo[3,2-d]pyrimidin-4-ol

[0834] A mixture of 7-bromo-4-methoxy-5-(4-phenoxyphenyl)-5H-pyrrolo[3,2-d]pyrimidine (1 g, 2.5 mmol) in MeOH (20 mL) and 6N HCl (20 mL) was refluxed overnight. Then it was cooled to room temperature and concentrated. The residue was neutralized with saturated NaHCO 3 and filtered. The filtered cake was dried to give the obtained product as a light yellow solid (850 mg, 88% yield). LCMS: calculated exact mass = 381.0; experimental value [M+H] + (ESI) = 382.2.

[0835] 7-Bromo-4-chloro-5-(4-phenoxyphenyl)-5H-pyrrolo[3,2-d]pyrimidine

[0836] A suspension of 7-bromo-5-(4-phenoxyphenyl)-5H-pyrrolo[3,2-d]pyrimidin-4-ol (850 mg, 2.2 mmol) in POCl 3 was stirred at 100 °C for 1.5 h. Then it was cooled to room temperature and concentrated, and the residue was quenched with saturated NaHCO 3 , extracted with DCM, dried and concentrated to afford the product as a light yellow solid (900 mg, 100% yield). LCMS: calculated exact mass = 399.0; experimental value [M+H] + (ESI) = 399.9.

[0837] 7-Bromo-5-(4-phenoxyphenyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine

[0838] To a suspension of 7-bromo-4-chloro-5-(4-phenoxyphenyl)-5H-pyrrolo[3,2-d]pyrimidine (900 mg, 2.24 mmol) in 1,4-dioxane (20 mL) was added NH 3 .H 2 O (20 mL), and the mixture was stirred overnight in a sealed tube at 130 °C. It was then cooled to room temperature and concentrated. The suspension was filtered to afford the product (900 mg, 100% yield). LCMS: calculated exact mass = 380.0; experimental value [M+H] + (ESI) = 380.8.

[0839] 5-(4-Phenoxyphenyl)-7-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine

[0840] To a mixture of 7-bromo-5-(4-phenoxyphenyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine (750 mg, 1.97 mmol), 4,4,5,5-tetramethyl-2-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)-1,3,2-dioxaborolane (2.6 g, 9.84 mol), Pd(dppf)Cl 2 (288 mg, 0.39 mmol) and Na 2 CO 3 (1.04 g, 9.84 mmol) in dioxane-H 2 O (40 mL - 4 mL) was heated at 100 °C under an Ar atmosphere for 4 h. After cooling to room temperature, the reaction mixture was concentrated and extracted with DCM. The organic layer was concentrated and purified by flash column chromatography (EtOAc, 100%) to give the product (50 mg, 6% yield). LCMS: calculated exact mass = 440.2; experimental value [M+H] + (ESI) = 440.9.

[0841] 5-(4-Phenoxyphenyl)-7-(1,4-dioxaspiro[4.5]dec-8-yl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine

[0842] To a solution of 5-(4-phenoxyphenyl)-7-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine (40 mg, 0.092 mmol) in EtOAc / MeOH (50 mL) was added Pd / C (20 mg, 10% in activated carbon). At room temperature in H 2Stir the mixture overnight under the atmosphere. Subsequently, filter and concentrate the filtrate to obtain the product (40 mg, 100% yield). LCMS: Calculated exact mass = 442.2; experimental value [M+H] + (ESI) = 442.8.

[0843] 4-(4-Amino-5-(4-phenoxyphenyl)-5H-pyrrolo[3,2-d]pyrimidin-7-yl)cyclohexan-1-one

[0844] To a mixture of 5-(4-phenoxyphenyl)-7-(1,4-dioxaspiro[4.5]dec-8-yl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine (50 mg, 0.11 mmol) in acetone (5 mL) was added 6N HCl in THF (2 mL). The mixture was stirred at 40 °C for 3 hours. Subsequently, it was cooled to room temperature, the mixture was neutralized with aqueous NaOH, extracted with DCM, dried and concentrated. The residue was purified by flash column chromatography (EtOAc, 100%) to give the product (15 mg, 30% yield). LCMS: Calculated exact mass = 398.17; experimental value [M+H] + (ESI) = 398.9.

[0845] 7-((cis)-4-(4-Methylpiperazin-1-yl)cyclohexyl)-5-(4-phenoxyphenyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine

[0846] and

[0847] 7-((trans)-4-(4-Methylpiperazin-1-yl)cyclohexyl)-5-(4-phenoxyphenyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine

[0848] To a mixture of 4-(4-amino-5-(4-phenoxyphenyl)-5H-pyrrolo[3,2-d]pyrimidin-7-yl)cyclohexan-1-one (15 mg, 0.038 mmol), 1-methylpiperazine (19 mg, 0.19 mmol) in DCE (5 mL) was added AcOH (1 drop). The reactants were stirred at room temperature for 2 hours. NaBH(OAc) 3 (64 mg, 0.304 mmol) was added portionwise. The reactants were stirred at room temperature overnight. The reactants were treated with saturated NaHCO 3Quench, extract with DCM, dry and concentrate. Purify the residue via Prep-TLC (DCM:MeOH = 10:1) to obtain the product 7-((trans)-4-(4-methylpiperazin-1-yl)cyclohexyl)-5-(4-phenoxyphenyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine (3 mg, 16% yield). LCMS: calculated exact mass = 482.3; experimental value [M+H] + (ESI) = 483.3; 1 H NMR (chloroform-d) δ 8.40 (s, 1H), 7.41 - 7.44 (m, 2H), 7.35 (d, J = 8.5 Hz, 2H), 7.19 - 7.25 (m, 1H), 7.11 (dd, J = 7.9, 6.1 Hz, 5H), 4.90 (br.s., 2H), 3.03 (br.s., 9H), 2.53 (br.s., 3H), 2.30 (d, J = 16.2 Hz, 4H), 1.60 - 1.71 (m, 6H); and 7-((cis)-4-(4-methylpiperazin-1-yl)cyclohexyl)-5-(4-phenoxyphenyl)-5H-pyrrolo[3,2-d]pyrimidin-4-amine (2 mg, 11% yield). LCMS: calculated exact mass = 482.3; experimental value [M+H] + (ESI) = 483.3; 1 H NMR (chloroform-d) δ 8.40 (s, 1H), 7.35 - 7.52 (m, 5H), 7.17 - 7.23 (m, 1H), 7.11 (t, J = 9.3 Hz, 4H), 4.82 - 5.01 (m, 2H), 3.00 (br.s., 6H), 2.54 (br.s., 3H), 2.03 (br.s., 2H), 1.87 (br.s., 2H), 1.77 (d, J = 10.7 Hz, 2H), 1.63 (d, J = 7.6 Hz, 2H).

[0849] HCK kinase assay: The HCK kinase reaction mixture (10 μL) containing 4 nM N-terminal GST-tagged HCK (75 - 526) purified from insect expression system, 5 μM control AMC substrate, 2 μM Src family kinase R110 substrate and in kinase reaction buffer (40 mM Tris-HCl pH 7.5, 20 mM MgCl 2 2, 0.1 mg / mL BSA, 1 mM MnCl 2, 50 μM ATP and test compounds starting at 1 μM and serially diluted 1:3 in (0.1 mM sodium vanadate) were incubated in a 384-well plate (Corning, catalog number 4514) at room temperature (22 - 25 °C) for 60 minutes. Subsequently, follow the procedure of the ProFluor Src Family Kinase Assay (Promega, catalog number V1271). Add 5 μL of protease solution to the reaction and incubate the mixture at room temperature (22 - 25 °C) for 60 minutes, followed by 5 μL of stabilizer solution. Read the fluorescence signal on an Envision multi-label reader (Perkin Elmer). Subsequently, read R110 at an excitation wavelength of 485 nm and an emission wavelength of 530 nm. Read the AMC signal at an excitation wavelength of 355 nm and an emission wavelength of 460 nm.

[0850] Description of HCK inhibitory activity

[0851]

[0852]

[0853] Table of HCK inhibitory activity

[0854]

[0855]

[0856]

[0857]

[0858] Evaluation of in vitro metabolic stability

[0859] Dog liver microsomes (0.5 mg / mL) were purchased from Corning. Stock solutions were prepared at 10 mM in DMSO for testing compounds. Aliquots of the stock solutions were diluted to 0.5 mM with acetonitrile and then further diluted to 1.5 μM after addition of liver microsomes / buffer. Aliquots of 30 μL of the 1.5 μM solution were mixed with 15 μL of 6 mM NADPH and the final concentration of NADPH was 2 mM, which had been pre-warmed to 37 °C. The final concentrations of the test compound and ketanserin were 1 μM. The plate was kept in a 37 °C water bath during the experiment duration. At each time point (0, 5, 15, 30, 45 minutes), 135 μL of acetonitrile was added to the corresponding wells. After quenching with acetonitrile at the final time point, the assay plate (IKA, MTS2 / 4) was shaken for 10 min (600 rpm / min) and then centrifuged at 5,594 g for 15 min (Thermo Multifuge × 3R). Aliquots of the supernatant were diluted 1:1 to distilled water and analyzed by LC-MS / MS. The peak area response ratio (PARR) of the compound to the internal standard at 5, 15, 30, 45 minutes was compared to the PARR at time 0 to determine the percentage of the test compound remaining at each time point. The half-life was calculated using Excel software and fitted with a monoexponential decay equation.

[0860] Evaluate in vivo pharmacokinetics

[0861] A total of 6 male non-juvenile Beagle dogs (Beijing Marshall Biotechnology Co., Ltd.) weighing approximately 8.00 - 10.00 kg were dosed at 1 mg / kg or 3 mg / kg by intravenous injection (iv) or oral gavage (po). The vehicles for iv and po were 100% normal saline and 100% (0.5% MC in water), respectively. IV administration was performed via the cephalic vein injection, and PO administration was performed via gavage. The animals were manually restrained, and approximately 0.5 mL of blood was collected from the cephalic vein / time point into pre-cooled K 2 EDTA tubes. The blood samples were placed on wet ice and centrifuged at 4 °C (2000 g, 5 min) to obtain plasma within 15 minutes of sample collection. The plasma samples were analyzed by UPLC / MS-MS. PK parameters were estimated using WinNonlin 6.4 by a non-compartmental model.

[0862] Evaluate the inhibitory activity of the following compounds against HCK and their circulating half-life (t 1 / 2 ) in dog LM. The compounds were synthesized as described above.

[0863]

[0864]

[0865]

[0866]

[0867] As shown in the above data table, the t of the compounds of Examples 127, 103, 165, 163, 19 and 21 1 / 2 is higher than that of the compound of Example 1. Although the half-life of the compound of Example 1 is 11 min, moving the piperazinyl-methyl one position in the compound of Example 19 unexpectedly makes the t 1 / 2 increase to 84 min. Replacing the pyrrole ring of Example 19 with a pyrazole ring in the compound of Example 21 further makes the t 1 / 2 increase to 128 min. Alternatively, replacing the pyrrole ring of the compound of Example 1 with a pyrazole ring and adding an ortho-fluoro substituent in the compound of Example 103 makes the t 1 / 2 increase to 58 min. Moving the piperazinyl-methyl of the compound of Example 103 one position in the compound of Example 127 further makes the t 1 / 2 increase to 118 min. Replacing the pyrazole ring of the compound of Example 127 with an imidazole in the compound of Example 165 gives the highest observed t 1 / 2 , 167 min. Moving the piperazinyl-methyl of the compound of Example 165 back to the terminal piperazine nitrogen in the compound of Example 163 makes the t 1 / 2 slightly decrease to 108 min. The longer t of Examples 103, 127 and 163 1 / 2 is confirmed by lower plasma clearance and higher in vivo bioavailability.

[0868] In addition, the compounds studied also show an unexpected effect of substituents on potency. For example, the compounds of Examples 127 and 21 differ only in the fluorine group, but the compound of Example 127 unexpectedly shows a potency nearly an order of magnitude higher. As another example, the compounds of Examples 127 and 17 differ only in the stereochemistry of the methyl group, but the compound of Example 127 shows a potency nearly three times greater.

Claims

1. A compound of formula (I): Wherein each dashed bond is independently a single bond or a double bond such that the bicyclic formed thereby is a heteroaryl; X is N; Q is C; Y is C and Z is N, or Y is N and Z is C; R 1 For W is C, R 2 is H, V is N, and T is N; R 3 selected from H and C 1 -C 6 a straight-chain or branched-chain alkyl group; R 3 ’ is H or C 1 -C 6 a straight-chain or branched-chain alkyl group; R 4 selected from H and C 1 -C 6 a straight-chain or branched-chain alkyl group; R 4 ’ is H or C 1 -C 6 a straight-chain or branched-chain alkyl group; R 5 is H; A 1 and A 2 are both CH 2 ; R 9 is F; R 10 is H; R 11 For B is O; K 1 and K 2 each is CH; R 14 is H; R 15 is H; and R 12 is H.

2. The compound according to claim 1, wherein R 1 is V is N; T is N; R 3 is C 1 -C 6 a straight-chain or branched-chain alkyl group; and R 4 is H or C 1 -C 6 a straight-chain or branched-chain alkyl group.

3. The compound according to claim 1, wherein R 1 is V is N; T is N; R 3 is C 1 -C 6 a straight-chain or branched-chain alkyl group; and R 4 is H or C 1 -C 6 a straight-chain or branched-chain alkyl group.

4. The compound according to claim 1, wherein R 3 is C 1 -C 6 a straight-chain or branched-chain alkyl group.

5. The compound according to claim 1, wherein the compound is selected from: or a pharmaceutically acceptable salt thereof.

6. Use of the compound of formula (I) according to claim 1 in the manufacture of a medicament for the treatment of acute myeloid leukemia in a subject in combination with a BCL-2 inhibitor.

7. The use according to claim 6, wherein the subject has FLT3-ITD+ acute myeloid leukemia.

8. The use according to claim 6, wherein the compound of formula (I) is an HCK inhibitor.

9. The use according to claim 6, wherein the medicament is for co-administration with an FLT3-ITD inhibitor.

10. The use according to claim 8, wherein the HCK inhibitor is a dual HCK / FLT3-ITD inhibitor.

11. The use according to claim 9, wherein the FLT3-ITD inhibitor is selected from AC220, sorafenib, PKC412, CEP-701, UNC2025, MLN518, KW-2449, and AMG-925, sunitinib, SU5614, AC2206, nilotinib, and PLX3397.

12. The use according to claim 8, wherein the BCL-2 inhibitor is selected from AT-101, TW-37, TM-1206, gossypol acetic acid, gossypol alcohol acid, apogossypol, apogossypolone, A385358, ABT-737, ABT-263, ABT-199, WEHI-539, BXI-61, BXI-72, obacalap, JY-1-106, and SAHB peptide.

13. The use according to claim 12, wherein the BCL-2 inhibitor is ABT-199.

14. The application according to claim 12, wherein the HCK inhibitor is and the BCL-2 inhibitor is ABT-199.

15. The application according to claim 12, wherein the HCK inhibitor is and the BCL-2 inhibitor is ABT-199.

16. The application according to claim 12, wherein the HCK inhibitor is and the BCL-2 inhibitor is ABT-199.

17. The application according to claim 12, wherein the HCK inhibitor is and the BCL-2 inhibitor is ABT-199.

18. The use according to claim 6, wherein the medicament is for co-administration with an anti-apoptotic agent.

19. The use according to claim 18, wherein the anti-apoptotic agent is retinoic acid, GGTI 298 TFA salt, BTSA1, AT406, SM-406, elisidepsin, STA-4783, galeterone, STA-9090, NQDI-1, or zoledronic acid.

20. The use according to claim 6, wherein the medicament is for co-administration with an Mcl-1 inhibitor.

21. The application according to claim 20, wherein the Mcl-1 inhibitor is 22. The use according to claim 6, wherein the medicament is for co-administration with an IAP antagonist.

23. The use according to claim 22, wherein the IAP antagonist is AT-406 / Debio 1143, GDC-0917 / CUDC-427, LCL161 or TL-32711.

24. The use according to claim 6, wherein the medicament is for co-administration with a cell cycle inhibitor.

25. The use according to claim 24, wherein the cell cycle inhibitor is selected from palbociclib, ribociclib, abemaciclib, flavopiridol, AT9283, alisertib, and MK-1775.

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