HK1210779B

HK1210779B - Inhibitors of bruton's tyrosine kinase

Info

Publication number: HK1210779B
Application number: HK15111588.8A
Authority: HK
Inventors: Niala Bhagirath; Romyr Dominique; Joshua Kennedy-Smith; Francisco Javier Lopez-Tapia; Eric Mertz; Qi Qiao; Sung-Sau So
Original assignee: F. Hoffmann-La Roche Ag
Priority date: 2012-10-26
Filing date: 2013-10-23
Publication date: 2017-12-15

Description

Tyrosine kinase inhibitors

Technical Field

The present invention relates to the use of novel compounds that inhibit Btk and can be used to treat autoimmune and inflammatory diseases caused by aberrant B-cell activation.

Background

Protein kinases constitute one of the largest families of human enzymes and regulate many different signaling processes by adding phosphate groups to proteins (T. Hunter, Cell 198750: 823-829). In particular, tyrosine kinases phosphorylate proteins on the phenol portion of tyrosine residues. The tyrosine kinase family includes members that control cell growth, migration, and differentiation. Abnormal kinase activity has been implicated in a number of human diseases, including cancer, autoimmune disorders and inflammatory diseases. Since protein kinases belong to key regulators of cell signaling, they provide the goal of regulating cellular function with small molecule kinase inhibitors and are therefore good drug design targets. In addition to the treatment of kinase-mediated disease processes, selective and potent inhibitors of kinase activity may also be useful in studying cellular signaling processes and in identifying other therapeutically significant cellular targets.

There is good evidence for the critical role of B cells in the pathogenesis of autoimmune and/or inflammatory diseases. Protein-based therapeutics that deplete B cells, such as Rituxan, are effective against autoantibody-induced inflammatory diseases such as rheumatoid arthritis (Rastetter et al Annu Rev Med 200455: 477). Therefore, inhibitors of protein kinases that play a role in B-cell activation should be useful therapeutic agents for B-cell mediated disease pathologies such as autoantibody production.

Signaling through the B-cell receptor (BCR) controls a range of B-cell responses, including proliferation and differentiation into mature antibody-producing cells. BCR is a key regulatory point for B-cell activity and aberrant signaling can lead to deregulated B-cell proliferation and the formation of pathogenic autoantibodies, which lead to a variety of autoimmune and/or inflammatory diseases. Tyrosine protein kinases (Btk) are non-BCR-related kinases that are proximal and immediately downstream of the membrane of BCR. The lack of Btk has been shown to block BCR signaling, and thus inhibition of Btk can be a useful therapeutic approach to block B cell-mediated disease processes.

Btk is a member of the Tec family of tyrosine kinases and has been shown to be a key regulator of early B-cell formation and mature B-cell activation and survival (Khan et al Immunity 19953: 283; Ellmeier et al J.exp.Med.2000192: 1611). Mutations in Btk in humans lead to the disorder X chromosome-linked gammoproteinemia (XLA) (reviewed in Rosen et al New Eng.J.Med.1995333: 431 and Lindval et al Immunol.Rev.2005203: 200). These patients are immunocompromised and show impaired B cell maturation, reduced immunoglobulin and peripheral B cell levels, reduced T cell-independent immune responses and impaired calcium mobilization following BCR stimulation.

Evidence for a role for Btk in autoimmune disorders and inflammatory diseases has been provided by Btk-deficient mouse models. In a preclinical murine model of Systemic Lupus Erythematosus (SLE), Btk deficient mice show significant improvement in disease progression. Furthermore, Btk-deficient mice are resistant to collagen-induced arthritis (Jansson and HolmdahlClin. exp. Immunol.199394: 459). Dose-dependent efficacy of selective Btk inhibitors in mouse models of arthritis has been demonstrated (z.pan et al, chem.med chem.20072: 58-61).

Btk is also expressed by cells other than B-cells that may be involved in disease processes. For example, Btk is expressed by mast cells and Btk-deficient bone marrow-derived mast cells exhibit impaired antigen-induced degranulation (Iwaki et al j.biol.chem.2005280: 40261). This shows that Btk can be used to treat pathological mast cell responses such as allergy and asthma. Furthermore, monocytes from XLA patients, where Btk activity is absent, show reduced TNF α production following stimulation (Horwood et al J Exp Med 197: 1603, 2003). Thus, TNF α -mediated inflammation can be modulated by small molecule Btk inhibitors. In addition, Btk has been reported to play a role in apoptosis (Islam and Smith immunol. rev.2000178: 49) and therefore Btk inhibitors would be useful for the treatment of certain B-cell lymphomas and leukemias (Feldhahn et al j.exp.med.2005201: 1837).

Summary of The Invention

The present application provides Btk inhibitor compounds of formula I, methods of use thereof, as described herein below:

the present application provides a compound of formula I,

wherein:

a is phenyl or piperidinyl;

R¹each independently of the others being halogen, lower alkyl, CH₂NHC(＝O)R^1’、CH₂N(CH₃)C(＝O)R^1’、CH₂NHC(＝O)CH₂NHR^1’、CH₂R¹' or CH₂NHR^1’；

n is 0, 1 or 2;

R^1’is phenyl, unsaturated or partially unsaturated bicyclic or monocyclic heteroaryl or heterocycloalkyl, optionally substituted with one or more R^1”Substitution;

R^1”each independently is lower alkyl, halogen, cycloalkyl, heterocycloalkyl, lower alkyl heterocycloalkyl, oxo, cyano lower alkyl, hydroxy lower alkyl or lower alkoxy;

R²is H, R³Or R⁴；

R³Is C (═ O) OR^3’、C(＝O)R^3’Or C (═ O) NH (CH)₂)₂R^3’；

R^3’Is H, lower alkyl, heterocycloalkyl, amino or OH;

R⁴is lower alkyl or heteroaryl, optionally substituted by one or moreR is^4’Substitution; and is

R^4’Is hydroxy, amino, OC (═ O) CH₂CH₃Or C (═ O) OH;

or a pharmaceutically acceptable salt thereof.

The present application provides a compound of formula I,

wherein:

a is phenyl or piperidinyl;

n is 0, 1 or 2;

R²is H, R³Or R⁴；

R³Is C (═ O) OR^3’、C(＝O)R^3’Or C (═ O) NH (CH)₂)₂R^3’；

R^3’Is H, lower alkyl, heterocycloalkyl, amino or OH;

R⁴is lowLower alkyl or heteroaryl, optionally substituted by one or more R^4’Substitution; and is

R^4’Is methyl, hydroxy, amino, CH₂-CH₂N(CH₃)₂、OC(＝O)CH₂CH₃、CH₂C(＝O)OH、CH₂CH₂OH or C (═ O) OH;

or a pharmaceutically acceptable salt thereof.

The present application provides methods for treating inflammatory and/or autoimmune disorders comprising administering to a patient in need thereof a therapeutically effective amount of a compound of formula I.

The present application provides pharmaceutical compositions comprising a compound of formula I in combination with at least one pharmaceutically acceptable carrier, excipient or diluent.

Detailed Description

Definition of

As used herein, the phrase "an" or "an" entity refers to one or more of that entity; for example, a compound refers to one or more compounds or at least one compound. Thus, the terms "a" (or "an)", "one or more" and "at least one" may be used interchangeably herein.

The phrase "as defined above" refers to the broadest definition of each group as provided in the summary of the specification or broadest claims. In all other embodiments provided below, substituents that may be present in each embodiment and are not explicitly defined remain with the broadest definition provided in the summary of the invention.

As used in this specification, the terms "comprises" and "comprising," whether in transitional phrases or in the main body of the claims, should be construed to have an open-ended meaning. That is, the term should be construed as synonymous with the phrase "having at least" or "including at least". When used in the context of a method, the term "comprising" means that the method includes at least the recited steps, but may include additional steps. The term "comprising" when used in the context of a compound or composition means that the compound or composition includes at least the recited features or components, but may also include additional features or components.

As used herein, unless specifically indicated otherwise, the use of the word "or" is in the "inclusive" sense of "and/or" and not in the "exclusive" sense of "or/and.

The term "independently" as used herein means that the variables are used in either case regardless of the presence or absence of variables having the same or different definitions within the same compound. Thus, in a compound where R "occurs twice and is defined as" independently carbon or nitrogen ", two R" can be carbon, two R "can be nitrogen, or one R" can be carbon and the other nitrogen.

When any variable occurs more than one time in any moiety or formula depicting or describing compounds employed or claimed in the present invention, its definition on each occurrence is independent of its definition at each other occurrence. Furthermore, combinations of substituents and/or variables are permissible only if such compounds result in stable compounds.

The symbol ". ang" at the end of a bond or "- - - - -" across a bond refers to the point of attachment of a functional group or other chemical moiety, respectively, to the remainder of the molecule of which it is a part. Thus, for example:

MeC(＝O)OR⁴whereinOr

A bond drawn into a ring system (as opposed to being attached at a distinct vertex) means that the bond can be attached to any suitable ring atom.

The term "optional" or "optionally" as used herein means that the subsequently described event or circumstance may, but need not, occur, and that the description includes instances where the event or circumstance occurs and instances where it does not. For example, "optionally substituted" means that the optionally substituted moiety may incorporate a hydrogen atom or a substituent.

The phrase "optional bond" means that the bond may or may not be present, and that the description includes single, double, or triple bonds. If a substituent is designated as "bond" or "absent," the atoms attached to that substituent are directly attached.

The term "about" as used herein means approximately, near. When the term "about" is used in conjunction with a numerical range, it modifies that range by extending the boundaries above and below the numerical values set forth. Generally, the term "about" as used herein changes a numerical value to a variance of 20% above and below the stated value.

Certain compounds of formula I may exhibit tautomerism. Tautomeric compounds may exist as two or more species that can be interconverted. Tautomers of proton transfer result from the migration of a covalently bound hydrogen atom between two atoms. Tautomers generally exist in equilibrium and attempts to isolate individual tautomers generally produce mixtures whose chemical and physical properties are consistent with mixtures of compounds. The position of equilibrium depends on the chemical characteristics within the molecule. For example, in the case of many aliphatic aldehydes and ketones, such as acetaldehyde, the keto form predominates, while in the phenols, the enol form predominates. Common tautomers of proton transfer include keto/enolAmide/imide acidAnd amidinesTautomers. The latter two are particularly common in heteroaryl and heterocyclic rings, and the present invention includes all tautomeric forms of the compounds.

Technical and scientific terms used herein have the meanings commonly understood by one of ordinary skill in the art to which this invention belongs, unless otherwise defined. Reference is made herein to various methods and materials known to those skilled in the art. Standard reference works listing general principles of pharmacology include Goodman and Gilman's The Pharmacological Basis of therapeutics, 10 th edition, McGraw Hill Companies Inc., new York (2001). Any suitable materials and/or methods known to those skilled in the art may be used in the practice of the present invention. However, preferred materials and methods are described. Materials, reagents, etc. referred to in the following description and examples are available from commercial sources unless otherwise indicated.

The definitions described herein may be appended to form chemically relevant combinations such as "heteroalkylaryl," "haloalkylheteroaryl," "arylalkyl heterocyclyl," "alkylcarbonyl," "alkoxyalkyl," and the like. When the term "alkyl" is used as a suffix following another term, as in "phenylalkyl" or "hydroxyalkyl", it is intended to indicate an alkyl group, as defined above, substituted with one to two substituents selected from the group of the additional specified nomenclature. Thus, for example, "phenylalkyl" refers to an alkyl group having 1 to 2 phenyl substituents, and thus includes benzyl, phenylethyl and biphenyl. "alkylaminoalkyl" is an alkyl group having 1 to 2 alkylamino substituents. "hydroxyalkyl" includes 2-hydroxyethyl, 2-hydroxypropyl, 1- (hydroxymethyl) -2-methylpropyl, 2-hydroxybutyl, 2, 3-dihydroxybutyl, 2- (hydroxymethyl), 3-hydroxypropyl and the like. Thus, as used herein, the term "hydroxyalkyl" is used to define a subset of heteroalkyl groups defined below. The term- (ar) alkyl refers to unsubstituted alkyl or aralkyl groups. The term (hetero) aryl or (hetero) aryl refers to aryl or heteroaryl.

The term "spirocycloalkyl" as used herein refers to a spiro cycloalkyl group such as, for example, spiro [3.3] heptane. The term spiroheterocycloalkyl as used herein refers to a spirocyclic heterocycloalkyl group such as, for example, 2, 6-diazaspiro [3.3] heptane.

The term "acyl" as used herein denotes a group of formula-C (═ O) R, wherein R is hydrogen or lower alkyl as defined herein. The term "alkylcarbonyl" as used herein, denotes a group of formula-C (═ O) R, wherein R is alkyl as defined herein. Term C_1-6Acyl means that the group-C (═ O) R contains 1 to 6 carbon atoms. The term "arylcarbonyl" as used herein denotes a group of formula C (═ O) R, wherein R is aryl; the term "benzoyl" as used herein is an "arylcarbonyl" group wherein R is phenyl.

The term "ester" as used herein denotes a group of formula-C (═ O) OR, wherein R is lower alkyl as defined herein.

The term "alkyl" as used herein denotes an unbranched or branched saturated monovalent hydrocarbon residue containing from 1 to 10 carbon atoms. The term "lower alkyl" denotes a straight or branched chain hydrocarbon residue containing 1 to 6 carbon atoms. "C" as used herein_1-10Alkyl "refers to an alkyl group consisting of 1 to 10 carbons. Examples of alkyl groups include, but are not limited to, lower alkyl groups including methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, tert-butyl or pentyl, isopentyl, neopentyl, hexyl, heptyl, and octyl.

When the term "alkyl" is used as a suffix following another term, as in "phenylalkyl" or "hydroxyalkyl", it is intended to indicate an alkyl group, as defined above, substituted with one to two substituents selected from the group of the additional specified nomenclature. Thus, for example, "phenylalkyl" denotes the group R 'R "-, where R' is phenyl and R" is alkylene as defined herein, with the understanding that the attachment point of the phenylalkyl moiety will be on the alkylene. Examples of arylalkyl groups include, but are not limited to, benzyl, phenylethyl, 3-phenylpropyl. The term "arylalkyl" or "aralkyl" is similarly construed, except that R' is aryl. The terms "(hetero) arylalkyl" or "(hetero) aralkyl" are similarly construed, except that R' is optionally aryl or heteroaryl.

The term "haloalkyl" or "halo-lower alkyl" or "lower haloalkyl" refers to a straight or branched chain hydrocarbon residue containing 1 to 6 carbon atoms, wherein one or more carbon atoms are substituted with one or more halogen atoms.

The term "alkylene (or alkenyl)" as used herein denotes a divalent saturated straight-chain hydrocarbon group of 1 to 10 carbon atoms (e.g., (CH)₂)_n) Or a branched saturated divalent hydrocarbon radical of 2 to 10 carbon atoms (e.g. -CHMe-or-CH)₂CH(i-Pr)CH₂-) unless otherwise indicated. Except in the case of methylene, the open valences of the alkylene groups are not attached to the same atom. Examples of alkylene groups include, but are not limited to: methylene, ethylene, propylene, 2-methyl-propylene, 1-dimethyl-ethylene, butylene, 2-ethylbutylene.

The term "alkoxy" as used herein denotes-O-alkyl, wherein alkyl is as defined above, such as methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, pentyloxy, hexyloxy, including isomers thereof. "lower alkoxy" as used herein denotes alkoxy having a "lower alkyl" group as defined above. "C" as used herein_1-10Alkoxy "means an-O-alkyl group wherein alkyl is C_1-10。

The term "PCy₃"refers to a phosphine trisubstituted with three cyclic moieties.

The term "haloalkoxy" or "halo-lower alkoxy" or "lower haloalkoxy" refers to lower alkoxy wherein one or more carbon atoms are substituted with one or more halogen atoms.

The term "hydroxyalkyl" as used herein denotes an alkyl group as defined herein wherein 1 to 3 hydrogen atoms on different carbon atoms are replaced by a hydroxyl group.

The terms "alkylsulfonyl" and "arylsulfonyl" as used herein refer to the formula-S (═ O)₂R, wherein R is independently alkyl or aryl, and alkyl and aryl are as defined herein. The term "heteroalkylsulfonyl" as used herein denotes the formula-S (═ O)₂The group of R, wherein R is "heteroalkyl" as defined herein.

The terms "alkylsulfonylamino" and "arylsulfonylamino" as used herein refer to the formula-NR' S (═ O)₂R, wherein R is independently alkyl or aryl, R' is hydrogen or C_1-3Alkyl, and alkyl and aryl are as defined herein.

The term "cycloalkyl" as used herein refers to a saturated carbocyclic ring containing 3 to 8 carbon atoms, i.e. cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl. "C" as used herein_3-7Cycloalkyl "refers to a cycloalkyl group consisting of 3 to 7 carbons in a carbocyclic ring.

The term carboxy-alkyl as used herein refers to an alkyl moiety in which one hydrogen atom has been replaced by a carboxy group, wherein it is understood that the point of attachment of the heteroalkyl group is through a carbon atom. The term "carboxy (or carboxyl)" means-CO₂And (4) a H part.

The term "heteroaryl" or "heteroaromatic" as used herein refers to a monocyclic or bicyclic group of 5 to 12 ring atoms having at least one aromatic or partially unsaturated ring, each ring containing 4 to 8 atoms, to which one or more N, O or S heteroatoms are attached, the remaining ring atoms being carbon, wherein it is understood that the attachment point of the heteroaryl group will be on the aromatic or partially unsaturated ring. As is well known to those skilled in the art, heteroaryl rings have less aromatic character than their all-carbon counterparts. Thus, for the purposes of the present invention, a heteroaryl group need only have some degree of aromatic character. Examples of heteroaryl moieties include monocyclic aromatic heterocycles having 5 to 6 ring atoms and 1 to 3 heteroatoms, including but not limited to pyridyl, pyrimidinyl, pyrazinyl, oxazinyl, pyrrolyl, pyrazolyl, imidazolyl, oxazolyl, 4, 5-dihydro-oxazolyl, 5, 6-dihydro-4H- [1, 3] oxazolyl, isoxazole, thiazole, isothiazole, triazoline, thiadiazole and oxadiazoline (oxadixoline), which may be optionally substituted with 1 or more, preferably 1 or 2, substituents selected from hydroxy, cyano, alkyl, alkoxy, thio, lower haloalkoxy, alkylthio, halogen, lower haloalkyl, alkylsulfinyl, alkylsulfonyl, halogen, amino, alkylamino, dialkylamino, aminoalkyl, alkylaminoalkyl and dialkylaminoalkyl, amino, alkylamino, dialkylamino, alkylthio, halogen, and pharmaceutically acceptable salts thereof, Nitro, alkoxycarbonyl and carbamoyl radicals, alkylcarbamoyl radicals, dialkylcarbamoyl radicals, arylcarbamoyl radicals, alkylcarbonylamino radicals and arylcarbonylamino radicals. Examples of bicyclic moieties include, but are not limited to, quinolinyl, isoquinolinyl, benzofuranyl, benzothienyl, benzoxazole, benzisoxazole, benzothiazole, naphthyridinyl, 5,6,7, 8-tetrahydro- [1, 6] naphthyridinyl, and benzisothiazole. The bicyclic moiety may be optionally substituted on either ring, but the point of attachment is on the ring containing the heteroatom.

The terms "heterocyclyl", "heterocycloalkyl" or "heterocycle" as used herein denote a monovalent saturated cyclic group consisting of one or more rings, preferably 1 to 2 rings (including spiro ring systems), each ring having 3 to 8 atoms to which one or more ring heteroatoms (selected from N, O or S (O))_0-2) And which may be optionally independently substituted by one or more, preferably 1 or 2, substituents selected from hydroxy, oxo, cyano, lower alkyl, lower alkoxy, lower haloalkoxy, alkylthio, halogen, lower haloalkyl, hydroxyalkyl, nitro, alkoxycarbonyl, amino, alkylamino, alkylsulfonyl, arylsulfonyl, alkylaminosulfonyl, arylaminosulfonylUnless otherwise indicated, alkyl, aryl, alkyl, aryl carbonyl amino and ionic forms thereof. Examples of heterocyclyl groups include, but are not limited to, morpholinyl, piperazinyl, piperidinyl, azetidinyl, pyrrolidinyl, hexahydroazepinylAlkyl, oxetanyl, tetrahydrofuryl, tetrahydrothienyl, oxazolidinyl, thiazolidinyl, isoxazolidinyl, tetrahydropyranyl, thiomorpholinyl, quinuclidinyl and imidazolinyl and ionic forms thereof. Examples may also be bicyclic, such as, for example, 3, 8-diaza-bicyclo [3.2.1]Octane, 2, 5-diaza-bicyclo [2.2.2]Octane or octahydro-pyrazino [2, 1-c][1，4]Oxazines.

Btk inhibitors

The present application provides a compound of formula I,

wherein:

a is phenyl or piperidinyl;

n is 0, 1 or 2;

R^1”each independently is lower alkyl, halogen, cycloalkyl, heterocycleAlkyl, lower alkyl heterocycloalkyl, oxo, cyano lower alkyl, hydroxy lower alkyl or lower alkoxy;

R²is H, R³Or R⁴；

R³Is C (═ O) OR^3’、C(＝O)R^3’Or C (═ O) NH (CH)₂)₂R^3’；

R^3’Is H, lower alkyl, heterocycloalkyl, amino or OH;

R⁴is lower alkyl or heteroaryl, optionally substituted by one or more R^4’Substitution; and is

or a pharmaceutically acceptable salt thereof.

Furthermore, it will be understood that reference to a particular residue A, R as disclosed herein is a reference to¹、R¹`、R¹``、R²、R³、R³`、R⁴And R^4`May be related to another residue A, R disclosed herein¹、R¹`、R¹``、R²、R³、R³`、R⁴And R^4`In combination with any other embodiment of (a).

The application provides compounds of formula I, wherein A is phenyl and R²Is H, and n is 1.

The present application provides compounds of formula I, wherein R¹Is halogen, R²Is H, and n is 1.

The present application provides compounds of formula I, wherein R¹Is a halogen.

The present application provides compounds of formula I, whichIn R²Is H, and n is 2.

The present application provides compounds of formula I, wherein R²Is H, n is 2, and one R¹Is a halogen.

The present application provides compounds of formula I, wherein R²Is H, n is 2, one R¹Is a lower alkyl group.

The present application provides compounds of formula I, wherein R¹Is CH₂NHC(＝O)R^1’。

The present application provides compounds of formula I, wherein R¹Is CH₂NHC(＝O)CH₂NHR^1’。

The present application provides compounds of formula I, wherein R¹Is CH₂NHR^1’。

The present application provides compounds of formula I, wherein R¹Is CH₂NHC(＝O)R^1’，R²Is H, and n is 1.

The present application provides compounds of formula I, wherein R¹Is CH₂NHC(＝O)CH₂NHR^1’，R²Is H, and n is 1.

The present application provides compounds of formula I, wherein R¹Is CH₂NHR^1’，R²Is H, and n is 1.

The application provides compounds of formula I, wherein n is 2, one R¹Is CH₂NHC(＝O)R^1’And R is²Is C (═ O) OR^3’、C(＝O)R^3’Or C (═ O) NH (CH)₂)₂R^3’。

The application provides compounds of formula I, wherein n is 2, one R¹Is CH₂NHC(＝O)R^1’And R is²Is lower alkyl or heteroaryl.

The present application provides compounds of formula I, wherein R^1’Is tert-butyl or halogen.

The present application provides compounds of formula I, wherein R^1’Is tert-butyl or halogen, R¹Is CH₂NHC(＝O)R^1’，R²Is H, and n is 1.

The application provides compounds of formula I, wherein one R¹Is fluorine, and R^1’Is a tert-butyl group.

The application provides compounds of formula I, wherein one R¹Is fluorine, and R^1’Is tert-butyl, n is 2, one R¹Is CH₂NHC(＝O)R^1’And R is²Is C (═ O) OR^3’、C(＝O)R^3’Or C (═ O) NH (CH)₂)₂R^3’。

The application provides compounds of formula I, wherein one R¹Is fluorine, and R^1’Is tert-butyl, n is 2, one R¹Is CH₂NHC(＝O)R^1’And R is²Is lower alkyl or heteroaryl.

The present application provides compounds of formula I, wherein a is piperidinyl.

The present application provides compounds of formula I, wherein a is piperidinyl, and n ═ 1.

The present application provides compounds of formula I, wherein a is piperidinyl, n ═ 1, and R¹Is CH₂NHC(＝O)R^1’。

The present application provides compounds of formula I, wherein a is piperidinyl, n ═ R¹Is CH₂NHC(＝O)R^1’And R is^1`Is phenyl, optionally substituted by one or more R^1``And (4) substitution.

The present application provides compounds of formula I, wherein a is piperidinyl, n ═ R¹Is CH₂NHC(＝O)R^1’And R is^1`Is phenyl, optionally substituted with one or more lower alkyl groups.

The present application provides compounds of formula I, wherein a is piperidinyl, n ═ R¹Is CH₂NHC(＝O)R^1’And R is^1`Is phenyl, optionally substituted with tert-butyl.

The present application provides compounds of formula I, wherein a is phenyl.

The present application provides compounds of formula I, wherein a is phenyl and n ═ 1 or 2.

The present application provides compounds of formula I wherein a is phenyl, n ═ 1 or 2, and one R¹Is CH₂NHC(＝O)R^1’。

The present application provides compounds of formula I wherein a is phenyl, n ═ 2, and one R¹Is CH₂NHC(＝O)R^1’And the other is halogen, and R^1`Is phenyl, unsaturated or partially unsaturated bicyclic or monocyclic heteroaryl or heterocycloalkyl, optionally substituted with one or more R^1``And (4) substitution.

The present application provides compounds of formula I wherein a is phenyl, n ═ 2, and one R¹Is CH₂NHC(＝O)R^1’And the other is F, and R^1`Is phenyl, optionally substituted by one or more R^1``And (4) substitution.

The present application provides compounds of formula I wherein a is phenyl, n ═ 2, and one R¹Is CH₂NHC(＝O)R^1’And the other is halogen, and R^1`Is phenyl, optionally substituted with one or more lower alkyl, halogen, cycloalkyl or heterocycloalkyl.

The present application provides compounds of formula I wherein a is phenyl, n ═ 2, and one R¹Is CH₂NHC(＝O)R^1’And the other is halogen, and R^1`Is phenyl, optionally substituted with one or more lower alkyl or halogen.

The present application provides compounds of formula I wherein a is phenyl, n ═ 2, and one R¹Is CH₂NHC(＝O)R^1’And the other is F, and R^1`Is phenyl, optionally substituted with one or more lower alkyl groups.

The present application provides compounds of formula I wherein a is phenyl, n ═ 2, and one R¹Is CH₂NHC(＝O)R^1’And the other is halogen, and R^1`Is phenyl, optionally substituted with one or more tert-butyl groups.

The present application provides compounds of formula I wherein a is phenyl, n ═ 2, and one R¹Is CH₂NHC(＝O)R^1’And the other is F, and R^1`Is phenyl, optionally substituted with one or more tert-butyl groups.

The present application provides compounds of formula I wherein a is phenyl, n ═ 2, and one R¹Is CH₂NHC(＝O)R^1’And the other is F, R^1`Is phenyl, optionally substituted with one or more tert-butyl groups, and R⁴Is heteroaryl, optionally substituted with one or more R^4`And (4) substitution.

The present application provides compounds of formula I wherein a is phenyl, n ═ 2, and one R¹Is CH₂NHC(＝O)R^1’And the other is F, R^1`Is phenyl, optionally substituted with one or more tert-butyl groups, and R⁴Is heteroaryl, optionally substituted with one or more methyl groups.

The present application provides compounds of formula I wherein a is phenyl, n ═ 2, and one R¹Is CH₂NHC(＝O)R^1’And the other is F, R^1`Is phenyl, optionally substituted with one or more tert-butyl groups, and R⁴Is heteroaryl, optionally substituted with methyl.

The present application provides compounds of formula I wherein a is phenyl, n ═ 2, and one R¹Is CH₂NHC(＝O)R^1’And the other is F, R^1`Is phenyl, optionally substituted by one or more tert-butyl groupsAnd R is⁴Is pyrazolyl optionally substituted by methyl.

The present application provides compounds of formula I, wherein a is phenyl, n ═ 1, R¹Is CH₂NHC(＝O)R^1’And R is^1`Is phenyl, optionally substituted with one or more tert-butyl groups.

The present application provides compounds of formula I, wherein a is phenyl, n ═ 1, R¹Is CH₂NHC(＝O)R^1’，R^1`Is phenyl, optionally substituted with one or more tert-butyl groups, and R⁴Is heteroaryl, optionally substituted with one or more R^4`And (4) substitution.

The present application provides compounds of formula I, wherein a is phenyl, n ═ 1, R¹Is CH₂NHC(＝O)R^1’，R^1`Is phenyl, optionally substituted with one or more tert-butyl groups, and R⁴Is pyrazolyl optionally substituted by one or more R^4`And (4) substitution.

The present application provides compounds of formula I, wherein a is phenyl, n ═ 1, R¹Is CH₂NHC(＝O)R^1’，R^1`Is phenyl, optionally substituted with one or more tert-butyl groups, and R⁴Is pyrazolyl, optionally substituted with one or more methyl groups.

The present application provides compounds of formula I, wherein a is phenyl, n ═ 1, R¹Is CH₂NHC(＝O)R^1’，R^1`Is phenyl, optionally substituted with one or more tert-butyl groups, and R⁴Is pyrazolyl optionally substituted by methyl.

The application provides compounds of formula I, wherein A is phenyl and R²Is H, n ═ 2, one R¹Is CH₂NHC(＝O)R^1’And the other is F, and R^1`Is phenyl, optionally substituted with one or more halogen, lower alkyl or cycloalkyl.

The present application provides a compound of formula I,wherein A is phenyl, R²Is H, n ═ 2, one R¹Is CH₂NHC(＝O)R^1’And the other is F, and R^1`Is phenyl, optionally substituted with one or more Cl, tert-butyl or cyclopropyl.

The present application provides compounds of formula I, wherein a is phenyl, n ═ 1, and R⁴(viii) heteroaryl, optionally substituted with one or more R^4`And (4) substitution.

The present application provides compounds of formula I, wherein a is phenyl, n ═ 2, and R⁴(viii) heteroaryl, optionally substituted with one or more R^4`And (4) substitution.

The present application provides compounds of formula I, wherein a is phenyl, n ═ 1, and R⁴(ix) heteroaryl, optionally substituted with one or more methyl groups.

The present application provides compounds of formula I, wherein a is phenyl, n ═ 2, and R⁴(ix) heteroaryl, optionally substituted with one or more methyl groups.

The application provides compounds of formula I, wherein A is phenyl and R²Is H, n is 2, and R is¹Is CH₂NHC(＝O)R^1’。

The application provides compounds of formula I, wherein A is phenyl and R²Is H, n ═ 2, one R¹Is CH₂NHC(＝O)R^1’And the other is halogen.

The application provides compounds of formula I, wherein A is phenyl and R²Is H, n ═ 2, one R¹Is CH₂NHC(＝O)R^1’And the other is F.

The application provides compounds of formula I, wherein A is phenyl and R²Is H, n ═ 2, one R¹Is CH₂NHC(＝O)R^1’And the other is F, and R^1`Is phenyl.

The present application provides compounds of formula I, wherein A is phenyl,R²is H, n ═ 2, one R¹Is CH₂NHC(＝O)R^1’And the other is F, R^1`Is phenyl, and R^1``Is lower alkyl or cycloalkyl.

The application provides compounds of formula I, wherein A is phenyl and R²Is H, n ═ 2, one R¹Is CH₂NHC(＝O)R^1’And the other is F, R^1`Is phenyl, and R^1``Is tert-butyl or cyclopropyl.

The application provides compounds of formula I, wherein A is phenyl and R²Is H, n ═ 2, one R¹Is CH₂NHC(＝O)R^1’And the other is F, R^1`Is phenyl, and R^1``Is a tert-butyl group.

The application provides compounds of formula I, wherein A is phenyl and R²Is H, n ═ 2, one R¹Is CH₂NHC(＝O)R^1’And the other is F, and R^1`Is an unsaturated or partially unsaturated monocyclic heteroaryl.

The application provides compounds of formula I, wherein A is phenyl and R²Is H, n ═ 2, one R¹Is CH₂NHC(＝O)R^1’And the other is F, and R^1`Is an unsaturated or partially unsaturated bicyclic heteroaryl.

The application provides compounds of formula I, wherein A is phenyl and R²Is H, n ═ 2, one R¹Is CH₂NHC(＝O)R^1’And the other is F, R^1`Is unsaturated or partially unsaturated monocyclic heteroaryl, and R^1``Is a lower alkyl group.

The application provides compounds of formula I, wherein A is phenyl and R²Is H, n ═ 2, one R¹Is CH₂NHC(＝O)R^1’And the other is F, R^1`Is an unsaturated or partially unsaturated bicyclic heteroaryl group, and R^1``Is a lower alkyl group.

This applicationPlease provide compounds of formula I, wherein A is phenyl and R²Is H, n ═ 2, one R¹Is CH₂NHC(＝O)R^1’And the other is F, R^1`Is an unsaturated or partially unsaturated monocyclic heteroaryl group, R^1``Is lower alkyl, and R⁴Is heteroaryl, optionally substituted with one or more R^4`And (4) substitution.

The application provides compounds of formula I, wherein A is phenyl and R²Is H, n ═ 2, one R¹Is CH₂NHC(＝O)R^1’And the other is F, R^1`Is an unsaturated or partially unsaturated bicyclic heteroaryl group, R^1``Is lower alkyl, and R⁴Is heteroaryl, optionally substituted with one or more R^4`And (4) substitution.

The application provides compounds of formula I, wherein A is phenyl and R²Is H, n ═ 2, one R¹Is CH₂NHC(＝O)R^1’And the other is F, R^1`Is an unsaturated or partially unsaturated monocyclic heteroaryl group, R^1``Is lower alkyl, and R⁴Is heteroaryl, optionally substituted with one or more methyl, hydroxy, amino, CH₂-CH₂N(CH₃)₂、OC(＝O)CH₂CH₃、CH₂C(＝O)OH、CH₂CH₂OH or C (═ O) OH.

The application provides compounds of formula I, wherein A is phenyl and R²Is H, n ═ 2, one R¹Is CH₂NHC(＝O)R^1’And the other is F, R^1`Is an unsaturated or partially unsaturated bicyclic heteroaryl group, R^1``Is lower alkyl, and R⁴Is heteroaryl, optionally substituted with one or more methyl groups.

The application provides compounds of formula I, wherein A is phenyl and R²Is H, n ═ 2, one R¹Is CH₂NHC(＝O)R^1’And the other is F, R^1`Is an unsaturated or partially unsaturated bicyclic heteroaryl group, R^1``Is lower alkyl, and R⁴Is heteroaryl optionally substituted with methyl.

The application provides compounds of formula I, wherein A is phenyl and R²Is H, n ═ 2, one R¹Is CH₂NHC(＝O)R^1’And the other is F, R^1`Is an unsaturated or partially unsaturated bicyclic heteroaryl group, R^1``Is lower alkyl, and R⁴Is pyrazolyl optionally substituted by methyl.

The application provides compounds of formula I, wherein A is phenyl and R²Is H, n is 1, and R is¹Is halogen or lower alkyl.

The application provides compounds of formula I, wherein A is phenyl and R²Is H, n is 1, and R is¹Is Cl, F or methyl.

The application provides compounds of formula I, wherein A is phenyl and R²Is H, and n ═ 2.

The application provides compounds of formula I, wherein A is phenyl and R²Is H, n is 2, and one R is¹Is halogen and the other is lower alkyl.

The application provides compounds of formula I, wherein A is phenyl and R²Is H, n is 2, and one R is¹Is Cl or F and the other is methyl.

The application provides compounds of formula I, wherein A is phenyl and R²Is H, n is 2, and two R are¹Are all methyl.

The present application provides a compound of formula I selected from:

4- (4-chloro-phenyl) -7H-pyrrolo [2,3-d ] pyrimidine;

4- (3-chloro-phenyl) -7H-pyrrolo [2,3-d ] pyrimidine;

4- (2-chloro-phenyl) -7H-pyrrolo [2,3-d ] pyrimidine;

4- (3-fluoro-4-methyl-phenyl) -7H-pyrrolo [2,3-d ] pyrimidine;

4- (2, 4-dimethyl-phenyl) -7H-pyrrolo [2,3-d ] pyrimidine;

4- (3, 4-dimethyl-phenyl) -7H-pyrrolo [2,3-d ] pyrimidine;

4-p-tolyl-7H-pyrrolo [2,3-d ] pyrimidine;

4- (3-chloro-4-methyl-phenyl) -7H-pyrrolo [2,3-d ] pyrimidine;

4-tert-butyl-N- [4- (7H-pyrrolo [2,3-d ] pyrimidin-4-yl) -benzyl ] -benzamide;

3-chloro-N- [4- (7H-pyrrolo [2,3-d ] pyrimidin-4-yl) -benzyl ] -benzamide;

2- (3-chloro-phenylamino) -N- [4- (7H-pyrrolo [2,3-d ] pyrimidin-4-yl) -benzyl ] -acetamide;

4-tert-butyl-N- [ 2-fluoro-4- (7H-pyrrolo [2,3-d ] pyrimidin-4-yl) -benzyl ] -benzamide;

4- {4- [ (4-tert-butyl-benzoylamino) -methyl ] -3-fluoro-phenyl } -7H-pyrrolo [2,3-d ] pyrimidine-6-carboxylic acid tert-butyl ester;

4- (4- ((4-tert-butylbenzoylamino) methyl) -3-fluorophenyl) -7H-pyrrolo [2,3-d ] pyrimidine-6-carboxylic acid;

4-tert-butyl-N- (2-fluoro-4- (6- (morpholine-4-carbonyl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl) benzyl) benzamide;

4- {4- [ (4-tert-butyl-benzoylamino) -methyl ] -3-fluoro-phenyl } -7H-pyrrolo [2,3-d ] pyrimidine-6-carboxylic acid dimethylamide;

4- {4- [ (4-tert-butyl-benzoylamino) -methyl ] -3-fluoro-phenyl } -7H-pyrrolo [2,3-d ] pyrimidine-6-carboxylic acid methylamide;

4- {4- [ (4-tert-butyl-benzoylamino) -methyl ] -3-fluoro-phenyl } -7H-pyrrolo [2,3-d ] pyrimidine-6-carboxylic acid (2-hydroxy-ethyl) -amide;

4- {4- [ (4-tert-butyl-benzoylamino) -methyl ] -3-fluoro-phenyl } -7H-pyrrolo [2,3-d ] pyrimidine-6-carboxylic acid (2-dimethylamino-ethyl) -amide;

4-tert-butyl-N- {1- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -piperidin-4-ylmethyl } -benzamide;

4-tert-butyl-N- {4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzyl } -benzamide;

4-cyclopropyl-N- {4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzyl } -benzamide;

4-isopropyl-N- {4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzyl } -benzamide;

n- {4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzyl } -4-oxetan-3-yl-benzamide;

4- (3-methyl-oxetan-3-yl) -N- {4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzyl } -benzamide;

4,5,6, 7-tetrahydro-benzo [ b ] thiophene-2-carboxylic acid 4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzylamide;

4-tert-butyl-N- { 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzyl } -benzamide;

6-tert-butyl-N- { 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzyl } -nicotinamide;

5-methyl-thiophene-2-carboxylic acid 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzylamide;

4-tert-butyl-N- (2-fluoro-4- {6- [1- (2-hydroxy-ethyl) -1H-pyrazol-4-yl ] -7H-pyrrolo [2,3-d ] pyrimidin-4-yl } -benzyl) -benzamide;

4-tert-butyl-N- { 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzyl } -N-methyl-benzamide;

5-methyl-thiophene-2-carboxylic acid { 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzyl } -methyl-amide;

2-tert-butyl-5- { 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzyl } -4, 5-dihydro-thieno [2,3-c ] pyrrol-6-one;

5-tert-butyl-isoxazole-3-carboxylic acid 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzylamide;

n- { 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzyl } -4- (3-methyl-oxetan-3-yl) -benzamide;

4- (cyano-dimethyl-methyl) -N- { 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzyl } -benzamide;

4,5,6, 7-tetrahydro-benzo [ b ] thiophene-2-carboxylic acid 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzylamide;

n- { 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzyl } -4- (1-hydroxy-1-methyl-ethyl) -benzamide;

3-tert-butyl-isoxazole-5-carboxylic acid 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzylamide

3-tert-butoxy-azetidine-1-carboxylic acid 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzylamide;

1, 3-dihydro-isoindole-2-carboxylic acid 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzylamide;

4-tert-butyl-N- (4- {6- [1- (2-dimethylamino-ethyl) -1H-pyrazol-4-yl ] -7H-pyrrolo [2,3-d ] pyrimidin-4-yl } -2-fluoro-benzyl) -benzamide;

3-tert-butoxy-azetidine-1-carboxylic acid 4- {6- [1- (2-dimethylamino-ethyl) -1H-pyrazol-4-yl ] -7H-pyrrolo [2,3-d ] pyrimidin-4-yl } -2-fluoro-benzylamide;

1, 3-dihydro-isoindole-2-carboxylic acid 4- {6- [1- (2-dimethylamino-ethyl) -1H-pyrazol-4-yl ] -7H-pyrrolo [2,3-d ] pyrimidin-4-yl } -2-fluoro-benzylamide;

[4- (4- {4- [ (4-tert-butyl-benzoylamino) -methyl ] -3-fluoro-phenyl } -7H-pyrrolo [2,3-d ] pyrimidin-6-yl) -pyrazol-1-yl ] -acetic acid ethyl ester;

[4- (4- {4- [ (4-tert-butyl-benzoylamino) -methyl ] -3-fluoro-phenyl } -7H-pyrrolo [2,3-d ] pyrimidin-6-yl) -pyrazol-1-yl ] -acetic acid;

n- (2-fluoro-4- (6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl) benzyl) -4,5,6, 7-tetrahydropyrazolo [1,5-a ] pyridine-2-carboxamide;

3-tert-butyl- [1,2,4] oxadiazole-5-carboxylic acid 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzylamide;

{ 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzyl } -carbamic acid tert-butyl ester; and

n- { 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzyl } -benzamide.

The present application provides methods of treating inflammatory and/or autoimmune disorders comprising administering to a patient in need thereof a therapeutically effective amount of a compound of formula I.

The present application provides a method of treating rheumatoid arthritis comprising administering to a patient in need thereof a therapeutically effective amount of a compound of formula I.

The present application provides a method of treating asthma comprising administering to a patient in need thereof a therapeutically effective amount of a compound of formula I.

The present application provides pharmaceutical compositions comprising compounds of formula I.

The present application provides the use of a compound of formula I for the manufacture of a medicament for the treatment of an inflammatory disorder.

The present application provides the use of a compound of formula I for the manufacture of a medicament for the treatment of autoimmune disorders.

The application provides the use of a compound of formula I for the manufacture of a medicament for the treatment of rheumatoid arthritis.

The present application provides the use of a compound of formula I for the manufacture of a medicament for the treatment of asthma.

The present application provides the use of a compound as described above in a medicament for the treatment of an inflammatory and/or autoimmune disorder.

The present application provides the use of a compound as described above in the treatment of rheumatoid arthritis.

The present application provides the use of a compound as described above in the treatment of asthma.

The present application provides compounds as described above for use in the treatment of inflammatory and/or autoimmune disorders.

The present application provides a compound as described above for use in the treatment of rheumatoid arthritis.

The present application provides a compound as described above for use in the treatment of asthma.

The present application provides a compound, method or composition as described above.

The present application provides methods of treating inflammatory and/or autoimmune disorders comprising administering to a patient in need thereof a therapeutically effective amount of a Btk inhibitor compound of formula I'.

The present application provides methods of treating arthritis comprising administering to a patient in need thereof a therapeutically effective amount of a Btk inhibitor compound of formula I'.

The present application provides methods of treating asthma comprising administering to a patient in need thereof a therapeutically effective amount of a Btk inhibitor compound of formula I'.

The present application provides a method of inhibiting B-cell proliferation comprising administering to a patient in need thereof a therapeutically effective amount of a Btk inhibitor compound of formula I'.

The present application provides methods of inhibiting Btk activity comprising administering a Btk inhibitor compound of any one of formula I', wherein the Btk inhibitor compound exhibits an IC of 50 micromolar or less in an in vitro biochemical assay of Btk activity₅₀。

In one variation of the above method, the Btk inhibitor compound exhibits an IC of 100 nanomolar or less in an in vitro biochemical assay of Btk activity₅₀。

In another variation of the above method, the compound exhibits an IC of 10 nanomolar or less in an in vitro biochemical assay of Btk activity₅₀。

The present application provides a method of treating an inflammatory disorder comprising co-administering to a patient in need thereof a therapeutically effective amount of an anti-inflammatory compound with a Btk inhibitor compound of formula I'.

The present application provides a method of treating arthritis comprising co-administering to a patient in need thereof a therapeutically effective amount of an anti-inflammatory compound with a Btk inhibitor compound of formula I'.

The present application provides for the treatment of lymphoma or BCR-ABL1⁺A method of leukemia cells by administering to a patient in need thereof a therapeutically effective amount of a Btk inhibitor compound of formula I'.

The present application provides pharmaceutical compositions comprising a Btk inhibitor compound of formula I' in combination with at least one pharmaceutically acceptable carrier, excipient, or diluent.

The present application provides the use of a compound of formula I' for the manufacture of a medicament for the treatment of an inflammatory disorder.

The present application provides the use of a compound of formula I' for the manufacture of a medicament for autoimmune disorders.

Compounds and preparation

Examples of representative compounds encompassed by the present invention and within the scope of the present invention are provided in the following table. These examples and preparations are provided so that those skilled in the art can more clearly understand and practice the present invention. They should not be considered as limiting the scope of the invention, but merely as being exemplary and representative thereof.

In general, the nomenclature used in this application is based on AUTONOMTM v.4.0, the Beilstein Institute computerized system for generating IUPAC systematic nomenclature. If there is an inconsistency between the structure being described and the name from which the structure was generated, the structure being described is given more weight. Further, if the stereochemistry of a structure or a moiety is not indicated using, for example, bold or dashed lines, the structure or moiety is to be interpreted as encompassing all stereoisomers of the structure or moiety.

Table I describes examples of compounds of formula I:

table I.

General synthetic schemes

The compounds of the present invention may be prepared by methods well known in the art. Suitable methods for synthesizing these compounds are provided in the examples. In general, the compounds of the present invention can be prepared according to one of the following synthetic routes (schemes 1-5). The starting materials are either commercially available or can be synthesized by methods well known to those skilled in the art.

Scheme 1

Compounds of formulae 5 and 6 of interest can be prepared according to scheme 1, where X is fluorine, hydrogen or methyl, and R is as described in the class of formula I above. The derivative 3 is obtained by the Suzuki aryl-aryl coupling reaction of the raw material of commercial 4-chloro-7-tosyl-7H-pyrrolo [2,3-d ] pyrimidine 1 and boric acid ester 2. Suzuki aryl-aryl coupling reaction conditions are reviewed in modern arene Chemistry 2002, 53-106. In carrying out the reaction, any of the conditions conventional in the Suzuki reaction may be used. In general, the Suzuki coupling reaction is carried out in the presence of a transition metal catalyst such as tetrakis (triphenylphosphine) palladium (0), a conventional organic solvent such as dimethoxyethane, and a weak inorganic base such as potassium carbonate. The reaction is carried out at a temperature of from room temperature to about 100 ℃ for from 1 hour to several hours, provided that conventional heating is employed. The reaction can also be carried out by microwave irradiation, which is generally carried out at higher temperatures (e.g. 160 ℃) but for shorter times (5-60 min). During the reaction, loss of tosyl was also observed. The tert-Butoxycarbonyl (BOC) protecting group in derivative 3 is readily removed under acidic conditions such as a mixture of trifluoroacetic acid (TFA) and Dichloromethane (DCM) to yield the free amine derivative 4. The reaction may be carried out at room temperature for a reaction time of 15 minutes to 3 hours. The coupling reaction between 4 and the carboxylic acid derivative can be carried out using standard peptide coupling reagents such as O- (7-azabenzotriazol-1-yl) -N, N' -tetramethyluronium hexafluoro-phosphate (HATU), conventional organic solvents such as N, N-Dimethylformamide (DMF), and bases such as Diisopropylethylamine (DIPEA) to give compounds such as 5. A list of coupling reagents that can also be used for this conversion can be determined in this Review (Chemical Review 2011, 111, 6557). The reaction may be carried out at room temperature for a reaction time of 1 hour to several hours. Alternatively, the free amine 4 can be coupled using 1, 1' -carbonyldiimidazole as a coupling reagent to produce urea derivatives, such as 6. The reaction can be carried out using DMF at a temperature of from room temperature to 90 ℃ for several hours. As is well known to those skilled in the art, other protecting Groups other than tosyl or BOC Groups can be used in this scheme (see Green's protective Groups in Organic Synthesis, Wiley and Sons, 2007, for the main references, p.g.m.wuts and t.w.greene).

Scheme 2

Compounds of formulae 13 and 14 of interest can be prepared according to scheme 2, where X is fluorine, hydrogen or methyl, and R is as described in the class of formula I above. Suzuki aryl-aryl couplings were performed as described in scheme 1. However, using a shorter reaction time (30 minutes) and heating at 160 ℃ under microwave irradiation, the tosyl protecting group can be maintained under those conditions. Bromination on C-2 of the pyrrolopyrimidine skeleton can be carried out using 1, 2-dibromo-tetrachloroethane in the presence of a strong base such as Lithium Diisopropylamide (LDA) to give derivative 9. The reaction may be carried out in an inert solvent such as Tetrahydrofuran (THF) at-78 deg.C for a reaction time of 2 hours to several hours (WO 2004/093812). Suzuki coupling between 9 and 10 can be performed using standard Suzuki conditions. The reaction was carried out at 160 ℃ under microwave irradiation for a longer reaction time (60 minutes), in which case the tosyl protecting group was also removed. The subsequent steps for the preparation of derivatives 13 and 14 are as described above.

Scheme 3

Scheme 3 describes the synthesis of such compounds as 23. Methylation at the nitrogen of carbamate 15 can be performed using a strong base such as sodium hydride (NaH) in the presence of methyl iodide and a polar solvent such as DMF. The reaction is carried out at 4 ℃ to room temperature for a reaction time of 2 hours to several hours. Palladium-catalyzed boration of carbamate 16 can be carried out using bis (pinacolato) diboron 17, a suitable source of palladium catalyst such as 1, 1' -bis (diphenylphosphino) ferrocene-palladium (II) dichloride and potassium acetate (Journal of Organic Chemistry 1995, 60, 7508-7510). The reaction may be carried out in a suitable solvent such as dioxane, DMF or NMP using conventional heating or microwave heating at a temperature of 90 ℃ to 150 ℃ for 1 hour to several hours. 4-chloro-6-iodo-7- (phenylsulfonyl) -7H-pyrrolo [2,3-d ] pyrimidine 19 can be coupled with boronic ester 10 using the Suzuki coupling conditions described above to provide derivative 20. Similarly, coupling of 18 and 20 was performed using the same standard conditions to give 21. The subsequent steps for the preparation of derivative 23 are as described above.

Scheme 4

Compounds of formula 33 of interest may be prepared according to scheme 4, wherein R is as defined for the class of formula I. The coupling partner 27 can be prepared in 2 steps from commercially available starting materials. As described above, standard coupling reagents can be used to form amide bonds in the derivative 26. Similarly, a borate functional group can also be introduced on compound 27 using standard conditions. The synthesis of derivative 30 has been described in the literature (WO 2011/149827). The Suzuki coupling between 27 and 30 can be carried out in a polar solvent such as DME, dioxane or DMF at a temperature of 60 ℃ to 100 ℃ using conventional heating methods for 1 hour to several hours. Microwave heating can considerably reduce the reaction time of Suzuki coupling (Current Organic Chemistry, 2010, 14, 1050-. Generally, only 10-60 minutes are required to complete the reaction. Deprotection and subsequent coupling of the BOC group on compound 31 to form an amide bond at 33 is as described above.

Scheme 5

Compounds of interest, e.g., 38, can be prepared according to scheme 5. The reaction between 19 and 36 can be carried out in a polar protic solvent such as ethanol in the presence of a base such as DIPEA or Triethylamine (TEA). The reaction may be carried out at 80 ℃ for a reaction time of 1 hour to several hours. Subsequent steps leading to the preparation of compound 38 are as described above.

Pharmaceutical compositions and administration

The compounds of the present invention may be formulated in a wide variety of oral administration dosage forms and carriers. Oral administration may be in the form of tablets, coated tablets, dragees, hard and soft gelatin capsules, solutions, emulsions, syrups or suspensions. The compounds of the present invention are effective when administered by other routes of administration, including sustained (intravenous drip) topical parenteral, intramuscular, intravenous, subcutaneous, transdermal (which may contain a penetration enhancer), buccal, nasal, inhalation, and suppository administration, among other routes of administration. The preferred mode of administration is generally oral using a conventional daily dosage regimen which may be adjusted according to the degree of affliction and the patient's response to the active ingredient.

The compound or compounds of the present invention, and their pharmaceutically acceptable salts, together with one or more conventional excipients, carriers or diluents, may be placed in the form of pharmaceutical compositions and unit dosages. The pharmaceutical compositions and unit dosage forms may comprise the usual ingredients in conventional proportions, with or without additional active compounds or principles, and may contain any suitable effective amount of the active ingredient commensurate with the intended daily dosage range of use. The pharmaceutical compositions may be used in solid form such as tablets or filled capsules, semi-solid, powder, sustained release formulations or liquid form such as solutions, suspensions, emulsions, elixirs or filled capsules for oral use; or in the form of suppositories for rectal or vaginal administration; or in the form of a sterile injectable solution for parenteral use. Typical formulations contain from about 5% to about 95% of the active compound or compounds (w/w). The term "formulation" or "dosage form" is intended to include both solid and liquid formulations of the active compound and those skilled in the art will understand that the active ingredient may be present in different formulations depending on the target organ or tissue and depending on the desired dosage and pharmacokinetic parameters.

The term "excipient" as used herein refers to a compound that is generally safe, non-toxic, and biologically or otherwise undesirable for use in preparing a pharmaceutical composition, and includes excipients that are useful for veterinary use as well as human pharmaceutical applications. The compounds of the invention may be administered alone, but will generally be administered in admixture with one or more suitable pharmaceutical excipients, diluents or carriers selected with regard to the intended route of administration and standard pharmaceutical practice.

By "pharmaceutically acceptable" it is meant that it can be used to prepare pharmaceutical compositions that are generally safe, non-toxic, and not biologically or otherwise undesirable, and includes that it is acceptable for veterinary as well as human pharmaceutical use.

The "pharmaceutically acceptable salt" form of the active ingredient may also initially impart desirable pharmacokinetic properties to the active ingredient which are lacking in the non-salt form and may even positively affect the pharmacodynamics of the active ingredient with respect to its in vivo therapeutic activity. The phrase "pharmaceutically acceptable salt" of a compound refers to a salt that is pharmaceutically acceptable and has the desired pharmacological activity of the parent compound. The salt comprises: (1) acid addition salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like; or with organic acids such as acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3- (4-hydroxybenzoyl) benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1, 2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphorsulfonic acid, 4-methylbicyclo [2.2.2] -oct-2-ene-1-carboxylic acid, glucoheptonic acid, 3-phenylpropionic acid, trimethylacetic acid, tert-butylacetic acid, laurylsulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, muconic acid, and the like; or (2) a salt formed when an acid proton present in the parent compound is replaced with a metal ion, such as an alkali metal ion, an alkaline earth ion, or an aluminum ion; or salts coordinated with organic bases such as ethanolamine, diethanolamine, triethanolamine, tromethamine, N-methylglucamine, etc.

Solid form preparations include powders, tablets, pills, capsules, cachets, suppositories, and dispersible granules. A solid carrier can be one or more substances that may also act as diluents, flavoring agents, solubilizers, lubricants, suspending agents, binders, preservatives, tablet disintegrating agents, or an encapsulating material. In powders, the carrier is typically a finely divided solid which is in admixture with the finely divided active ingredient. In tablets, the active ingredient is usually mixed with a carrier having the necessary binding capacity in suitable proportions and compacted in the shape and size desired. Suitable carriers include, but are not limited to, magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, a low melting wax, cocoa butter, and the like. Formulations in solid form may contain, in addition to the active ingredient, colorants, flavors, stabilizers, buffers, artificial and natural sweeteners, dispersants, thickeners, solubilizing agents, and the like.

Liquid formulations are also suitable for oral administration, including liquid formulations including emulsions, syrups, elixirs, aqueous solutions, aqueous suspensions. These include solid form preparations which are intended to be converted, shortly before use, to liquid form preparations. Emulsions may be prepared in the form of solutions, for example in the form of aqueous propylene glycol solutions, or may contain emulsifying agents, for example lecithin, sorbitan monooleate, or acacia. Aqueous solutions may be prepared by dissolving the active ingredient in water and adding suitable colorants, fragrances, stabilizers and thickeners. Aqueous suspensions may be formulated by dispersing the finely divided active ingredient in water together with viscous materials such as natural or synthetic gums, resins, methylcellulose, sodium carboxymethylcellulose and other well-known suspending agents.

The compounds of the invention may be formulated for parenteral administration (e.g., by injection, e.g., bolus injection, or continuous infusion) and may be presented in unit dosage form in ampoules, pre-filled syringes, small volume infusion solutions or in multi-dose containers with an added preservative. The compositions may take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, for example, solutions in aqueous polyethylene glycol. Examples of oily or nonaqueous carriers, diluents, solvents or vehicles include propylene glycol, polyethylene glycol, vegetable oils (e.g., olive oil), and injectable organic esters (e.g., ethyl oleate), and may contain formulatory agents such as preservatives, wetting agents, emulsifying or suspending agents, stabilizing agents and/or dispersing agents. Alternatively, the active ingredient may be presented in powder form, obtained by aseptic isolation of sterile solid or by lyophilization from solution, for constitution with a suitable vehicle, e.g., sterile, pyrogen-free water, before use.

The compounds of the present invention may be formulated for topical administration to the epidermis as an ointment, cream or lotion, or as a transdermal patch. Ointments and creams may, for example, be formulated with an aqueous or oily base with the addition of suitable thickening and/or gelling agents. Lotions may be formulated with an aqueous or oily base and will in general also contain one or more emulsifying agents, stabilizing agents, dispersing agents, suspending agents, thickening agents, or coloring agents. Formulations suitable for topical administration in the mouth include: lozenges comprising the active ingredient in a flavoured base, usually sucrose and acacia or tragacanth; pastilles comprising the active ingredient in an inert base such as gelatin and glycerin or sucrose and acacia; and mouthwashes comprising the active ingredient in a suitable liquid carrier.

The compounds of the present invention may be formulated for administration as suppositories. A low melting wax, such as a mixture of fatty acid glycerides or cocoa butter, is first melted and the active ingredient is dispersed homogeneously, for example by stirring. The molten homogeneous mixture is then poured into a suitably sized mold, allowed to cool and solidify.

The compounds of the present invention may be formulated for vaginal administration. Pessaries, tampons, creams, gels, pastes, foams or sprays containing in addition to the active ingredient such carriers as are known in the art to be suitable.

The compounds of the present invention may be formulated for nasal administration. The solution or suspension is applied directly to the nasal cavity by conventional means, for example with a dropper, pipette or nebulizer. The formulations may be provided in single or multiple dose forms. In the latter case of a dropper or pipette, this can be achieved by the patient administering an appropriate, predetermined volume of solution or suspension. In the case of a nebulizer, this can be achieved by, for example, a metered nebulization spray pump.

The compounds of the invention may be formulated for aerosol administration, particularly to the respiratory tract, and include intranasal administration. The compounds typically have small particle sizes, for example about five (5) microns or less. Such particle sizes may be obtained by means known in the art, for example by micronization. The active ingredient is provided in a pressurized container with a suitable propellant, such as a chlorofluorocarbon (CFC), for example dichlorodifluoromethane, trichlorofluoromethane or dichlorotetrafluoroethane, or carbon dioxide or other suitable gas. The aerosol may also conveniently contain a surfactant such as lecithin. The dosage of the medicament may be controlled by a metering valve. Alternatively, the active ingredient may be provided in the form of a dry powder, for example a powder mix of the compound in a suitable powder base such as lactose, starch derivatives such as hydroxypropylmethyl cellulose and polyvinylpyrrolidone (PVP). The powder carrier will form a gel in the nasal cavity. The powder compositions may be presented in unit dosage form, for example in capsules or cartridges of, for example, gelatin, or in blister packs, from which the powder may be administered by means of an inhaler.

When desired, the formulations can be prepared with an enteric coating suitable for sustained or controlled release administration of the active ingredient. For example, the compounds of the present invention may be formulated in a transdermal or subcutaneous drug delivery device. These delivery systems are advantageous when sustained release of the compound is desired and when patient compliance with a treatment regimen is important. Compounds in transdermal delivery systems are often attached to a skin-adherent solid support. The compound of interest may also be combined with a penetration enhancer such as lauryl nitrogenA ketone (1-dodecylaza-cycloheptan-2-one) combination. The sustained release delivery system is inserted subcutaneously into the subcutaneous layer by surgery or injection. Subcutaneous implants encapsulate compounds in a lipid soluble film such as silicone rubber or a biodegradable polymer such as polylactic acid (polylactic acid).

In Remington: suitable formulations are described in pharmaceutical Science and Practice (Remington: The Science and Practice of Pharmacv1995, edited by e.w. martin, Mack publishing company, 19 th edition, Easton, pa) together with pharmaceutical carriers, diluents and excipients. The skilled formulation scientist is able to modify the formulation within the scope of the present specification to provide a variety of formulations for the specific route of administration without destabilizing the compositions of the present invention or compromising their therapeutic activity.

Modifications that increase the solubility of the compounds of the invention in water or other excipients, for example, can be readily achieved by minor modifications (salt formation, esterification, etc.), which are within the ordinary skill in the art. It is also within the ordinary skill in the art to vary the route of administration and dosage regimen of a particular compound in order to maximize the beneficial effect of the pharmacokinetics of the compounds of the present invention in a patient.

As used herein, the term "therapeutically effective amount" refers to the amount required to alleviate the symptoms of a disease in an individual. The dosage will be adjusted to the individual need in each particular case. The dosage may vary within a wide range depending on a number of factors such as the severity of the disease to be treated, the age and general health of the patient, other drugs to treat the patient, the route and form of administration, and the preference and experience of the practitioner involved. For oral administration, daily dosages of between about 0.01 and about 1000mg/kg body weight per day should be appropriate in the case of monotherapy and/or in the case of combination therapy. Preferred daily dosages are between about 0.1 and about 500mg/kg body weight per day, more preferably between 0.1 and about 100mg/kg body weight per day, and most preferably between 1.0 and about 10mg/kg body weight per day. Thus, for administration to a 70kg human, the dosage range is about 7mg to 0.7 g/day. The daily dose may be administered as a single dose or in divided doses, typically in the range of 1-5 doses per day. In general, treatment is initiated with smaller doses that are less than the optimal dose of the compound. Subsequently, the dose is increased by small increments until the optimum effect is achieved for the individual patient. One of ordinary skill in the art of treating the diseases described herein will be able to determine, without undue experimentation and based on personal knowledge and experience and the disclosure of this application, a therapeutically effective amount of a compound of the present invention for a given disease and patient.

The pharmaceutical preparation is preferably in unit dosage form. In this form, the preparation is subdivided into unit doses containing appropriate quantities of the active ingredient. The unit dosage form may be a packaged preparation, the package containing discrete quantities of preparation, such as tablets, capsules, and powders in vials or ampoules. In addition, the unit dosage form can be a capsule, tablet, cachet, or lozenge itself, or it can be the appropriate number of any of these in packaged form.

Indications and treatment methods

The compounds of formula I inhibit tyrosine kinases (Btk). Upstream kinase activation of Btk results in phospholipase-C γ activation, thereby stimulating proinflammatory mediator release. The compounds of formula I are useful in the treatment of arthritis and other anti-inflammatory and autoimmune diseases. Accordingly, the compounds of formula I are useful for the treatment of arthritis. The compounds of formula I are useful for inhibiting Btk in cells and modulating B-cell development. The invention also encompasses pharmaceutical compositions comprising a compound of formula I in combination with a pharmaceutically acceptable carrier, excipient or diluent.

The compounds described herein are kinase inhibitors, particularly Btk inhibitors. These inhibitors are useful for treating one or more diseases responsive to kinase inhibition, including diseases responsive to Btk inhibition and/or B-cell proliferation, in a mammal. Without wishing to be bound by any particular theory, it is believed that the interaction of the compounds of the present invention with Btk results in the inhibition of Btk activity and thus brings about the pharmaceutical utility of these compounds. Accordingly, the invention includes methods of treating a mammal, e.g., a human, suffering from a disease responsive to inhibition of Btk activity and/or B-cell proliferation comprising administering to a mammal having such a disease an effective amount of at least one chemical entity provided herein. Effective concentrations can be determined experimentally, for example by measuring the blood concentration of the compound, or theoretically, providing a calculated bioavailability. Other kinases that may be affected in addition to Btk include, but are not limited to, other tyrosine kinases and serine/threonine kinases.

Kinases play a significant role in the signaling pathways that control basic cellular processes such as proliferation, differentiation and death (apoptosis). Abnormal kinase activity has been implicated in a variety of diseases including a variety of cancers, autoimmune and/or inflammatory diseases, and acute inflammatory responses. The multifaceted role of kinases in key cellular signaling pathways provides significant opportunities to identify new drugs that target kinases and signaling pathways.

One embodiment includes a method of treating a patient having an autoimmune and/or inflammatory disease or an acute inflammatory response in response to inhibition of Btk activity and/or B cell proliferation.

Autoimmune and/or inflammatory diseases that may be affected using the compounds and compositions according to the invention include, but are not limited to: psoriasis, allergies, crohn's disease, irritable bowel syndrome, sjogren's disease, tissue graft rejection and hyperacute rejection of transplanted organs, asthma, systemic lupus erythematosus (and associated glomerulonephritis), dermatomyositis, multiple sclerosis, scleroderma, vasculitis (ANCA-associated and other vasculitis), autoimmune hemolytic and thrombocytopenic symptoms, goodpasture's syndrome (and associated glomerulonephritis and pulmonary hemorrhage), atherosclerosis, rheumatoid arthritis, chronic Idiopathic Thrombocytopenic Purpura (ITP), addison's disease, parkinson's disease, alzheimer's disease, diabetes, septic shock and myasthenia gravis.

Included herein are methods of treatment wherein at least one chemical entity provided herein is administered in combination with an anti-inflammatory agent. Anti-inflammatory agents include, but are not limited to: NSAIDs, nonspecific and COX-2 specific cyclooxygenase enzyme inhibitors, gold compounds, corticosteroids, methotrexate, tumor necrosis factor receptor (TNF) receptor antagonists, immunosuppressive agents, and methotrexate.

Examples of NSAIDs include, but are not limited to, ibuprofen, flurbiprofen, naproxen and naproxen sodium, diclofenac, a combination of diclofenac sodium and misoprostol, sulindac, oxaprozin, diflunisal, piroxicam, indomethacin, etodolac, fenoprofen calcium, ketoprofen, nabumetone sodium, sulfasalazine, tolmetin sodium, and hydroxychloroquine. Examples of NSAIDs also include COX-2 specific inhibitors such as celecoxib, valdecoxib, lumiracoxib, and/or etoricoxib.

In some embodiments, the anti-inflammatory agent is a salicylate. Salicylates include, but are not limited to, acetylsalicylic acid or aspirin, sodium salicylate, and choline salicylate and magnesium salicylate.

The anti-inflammatory agent may also be a corticosteroid. For example, the corticosteroid can be cortisone, dexamethasone, methylprednisolone, prednisolone sodium phosphate, or prednisone.

In further embodiments, the anti-inflammatory agent is a gold compound such as gold sodium thiomalate or auranofin.

The invention also includes embodiments wherein the anti-inflammatory agent is a metabolic inhibitor such as a dihydrofolate reductase inhibitor, e.g., methotrexate, or a dihydroorotate dehydrogenase inhibitor, e.g., leflunomide.

Other embodiments of the present invention relate to combinations wherein at least one anti-inflammatory compound is an anti-C5 monoclonal antibody (such as eculizumab or peclizumab), a TNF antagonist such as etanercept or infliximab, which is an anti-TNF α monoclonal antibody.

Other embodiments of the invention relate to combinations wherein at least one of the active agents is an immunosuppressant compound such as an immunosuppressant compound selected from the group consisting of methotrexate, leflunomide, cyclosporin, tacrolimus, azathioprine and mycophenolate mofetil.

BTK-expressing B-cells and B-cell precursors have been implicated in the pathology of B-cell malignancies, including but not limited to B-cell lymphomas, lymphomas (including hodgkin and non-hodgkin lymphomas), hairy cell lymphomas, multiple myeloma, chronic and acute myeloid leukemias, and chronic and acute lymphocytic leukemias.

BTK has been shown to be an inhibitor of the Fas/APO-1(CD-95) death-inducing signaling complex (DISC) in B-lineage lymphoid cells. The fate of leukemia/lymphoma cells may lie in the balance between the reverse pro-apoptotic role of caspases activated by DISC and the upstream anti-apoptotic regulatory mechanisms including BTK and/or its substrates (Vassilev et al, j.biol.chem.1998, 274, 1646-1656).

It has also been found that BTK inhibitors can be used as chemosensitizers and thus in combination with other chemotherapeutic agents, in particular agents that induce apoptosis. Examples of other chemotherapeutic agents that may be used in combination with the chemosensitizing BTK inhibitor include topoisomerase I inhibitors (camptothecin or topotecan), topoisomerase II inhibitors (such as daunomycin and etoposide), alkylating agents (such as cyclophosphamide, melphalan and BCNU), tubulin-directed drugs (such as taxol and vinblastine), and biologicals (e.g., antibodies such as anti-CD 20 antibodies, IDEC 8, immunotoxins and cytokines).

Btk activity is also associated with some leukemias that express a bcr-abl fusion gene resulting from translocation of part of chromosomes 9 and 22. This abnormality is commonly observed in chronic myelogenous leukemia. Btk is constitutively phosphorylated by bcr-abl kinase, which triggers a downstream survival signal that prevents apoptosis in bcr-abl cells (N.Feldhahn et al J.exp.Med.2005201 (11): 1837. 1852).

Method of treatment

The present application provides methods of treating inflammation comprising administering to a patient in need thereof a therapeutically effective amount of a compound of formula I.

The present application provides methods of treating inflammatory and/or autoimmune disorders comprising administering to a patient in need thereof a therapeutically effective amount of a Btk inhibitor compound of formula I.

The present application provides methods of treating arthritis comprising administering to a patient in need thereof a therapeutically effective amount of a Btk inhibitor compound of formula I.

The present application provides methods of treating asthma comprising administering to a patient in need thereof a therapeutically effective amount of a Btk inhibitor compound of formula I.

The present application provides a method of inhibiting B-cell proliferation comprising administering to a patient in need thereof a therapeutically effective amount of a Btk inhibitor compound of formula I.

The present application provides methods of inhibiting Btk activity comprising administering a Btk inhibitor compound of any one of formula I, wherein the Btk inhibitor compound exhibits an IC of 50 micromolar or less in an in vitro biochemical assay of Btk activity₅₀。

In another variation of the above method, the Btk inhibitor is a pharmaceutically acceptable salt, solvate, or prodrug thereofThe compounds exhibit an IC of 10 nanomolar or less in an in vitro biochemical assay of Btk activity₅₀。

The present application provides methods of treating an inflammatory disorder comprising co-administering to a patient in need thereof a therapeutically effective amount of an anti-inflammatory compound with a Btk inhibitor compound of formula I.

The present application provides methods of treating arthritis comprising co-administering to a patient in need thereof a therapeutically effective amount of an anti-inflammatory compound with a Btk inhibitor compound of formula I.

The present application provides for the treatment of lymphoma or BCR-ABL1⁺A method of leukemia cells by administering to a patient in need thereof a therapeutically effective amount of a Btk inhibitor compound of formula I.

Examples

General conditions

The compounds of the present invention can be prepared starting from commercially available starting materials by using conventional synthetic techniques and procedures well known to those skilled in the art. The following summary is a reaction scheme suitable for preparing such compounds. Further examples may be determined in particular embodiments.

Preparation examples

Specific abbreviations

Detailed description of the general experiments

Reagents were purchased from Aldrich, Oakwood, Matrix or other suppliers and used without further purification. The reaction using microwave irradiation for heating was carried out using a Personal Chemistry emerys Optimizer System or a CEMDiscovery System. Purification on a multi-milligram to multi-gram scale is carried out by methods known to those skilled in the art, such as silica gel column elution; preparative flash column purification was also performed in some cases by experimental disposable pre-packed multi-gram silica gel column (RediSep) eluted with CombiFlash system. Biotage^TMAnd ISCO^TMAlso a rapid column apparatus which can be used for the purification of intermediates of the invention.

For the purpose of judging the identity and purity of the compound, LC/MS (liquid chromatography/mass spectrometry) spectra were recorded using the following system. For the determination of the mass spectrum, the system consists of a Micromass Platform II spectrophotometer: positive mode ES ionization (mass range: 150-. The simultaneous chromatographic separations were performed using the following HPLC system: ES industries Chromegabond WR C-183u(3.2x 30mm) column; mobile phase A: water (0.02% TFA) and phase B: acetonitrile (0.02% TFA); gradient 10% B-90% B, 3 min; equilibration time 1 minute; the flow rate was 2 mL/min.

Many of the compounds of formula 1 are also purified by reverse phase HPLC using methods well known to those skilled in the art. In some cases, preparative HPLC purification was performed using PE Sciex 150EX Mass Spec, which controls a Gilson 215 harvester connected to a Shimadzu preparative HPLC system and a Leap autoinjector. Compound was collected from the elution stream using LC/MS detection in cation detection: the compound was eluted from the C-18 column (20 mL/min 2.0X 10 cm) using a suitable linear gradient profile over 10 min, solvent (A) 0.05% TFA/H₂O and solvent (B) 0.035% TFA/acetonitrile. For injection on an HPLC system, the crude sample was dissolved in a mixture of methanol, acetonitrile and DMSO.

By passing¹H-NMR, the compound was characterized using a Bruker 400MHz NMR spectrophotometer.

The compounds of the invention can be synthesized according to known techniques. The following examples and references are provided to aid in the understanding of the present invention. However, these examples are not intended to limit the invention, the exact scope of which is set forth in the appended claims. The names of the end products in the examples were generated using Isis AutoNom 2000.

Preparation examples

Example I-1

4- (4-chloro-phenyl) -7H-pyrrolo [2,3-d ] pyrimidine

4-chloro-7H-pyrrolo [2,3-d ] is combined in 2mL of water in a 10mL microwave tube]Pyrimidine (200mg, 1.3mmol, Eq: 1.00), 4-chlorophenylboronic acid (204mg, 1.3mmol, Eq: 1.00) and potassium carbonate (720mg, 5.21mmol, Eq: 4.00) with DME (4.00 ml). Adding Pd (PPh)₃)₄(78mg, 0.068 mmol). The reaction mixture was irradiated in a microwave at 160 ℃ for 60 minutes. The resulting solution was diluted with EtOAc and washed with water. The combined organic phases were dried over anhydrous sodium sulfate and then evaporated. The crude material was dissolved with DCM and filtered. The title compound was obtained as a green solid (90mg, 30% yield). LC/MS: calculated m/z value C₁₂H₈ClN₃([M+H]⁺): 230.6, measurement: 230.1.

example I-2

4- (3-chloro-phenyl) -7H-pyrrolo [2,3-d ] pyrimidine

The title compound was obtained in a similar manner to example 1 using 3-chlorophenylboronic acid.

Example I-3

4- (2-chloro-phenyl) -7H-pyrrolo [2,3-d ] pyrimidine

The title compound was obtained in a similar manner to example 1 using 2-chlorophenylboronic acid.

Example I-4

4- (3-fluoro-4-methyl-phenyl) -7H-pyrrolo [2,3-d ] pyrimidine

The title compound was obtained in a similar manner to example 1 using 3-fluoro-4-methylphenylboronic acid.

Examples I to 5

4- (2, 4-dimethyl-phenyl) -7H-pyrrolo [2,3-d ] pyrimidine

4-chloro-7-tosyl-7H-pyrrolo [2,3-d ] combined in 2mL of water in a 10mL sealable microwave tube]Pyrimidine (210 mg)0.681mmol, Eq: 1.00), 2, 4-dimethylphenylboronic acid (112mg, 0.749mmol, Eq: 1.1) and potassium carbonate (376mg, 2.72mmol, Eq: 4.00) with DME (4 mL). Adding Pd (PPh)₃)₄(79mg, 0.068mmol, Eq: 0.1). The reaction mixture was heated in a microwave at 160 ℃ for 60 minutes. The resulting solution was diluted with EtOAc and washed with brine. The combined organic phases were dried over anhydrous sodium sulfate and the solvent was removed under reduced pressure. The crude material was purified by column chromatography (silica gel, 5-70% ethyl acetate in hexane).

The title compound was obtained as a solid (51mg, 34% yield). LC/MS: calculated m/z value C₁₄H₁₃N₃([M+H]⁺): 224.2, measurement: 224.2.

examples I to 6

4- (3, 4-dimethyl-phenyl) -7H-pyrrolo [2,3-d ] pyrimidine

4-chloro-7-tosyl-7H-pyrrolo [2,3-d ] combined in 2mL of water in a 10mL sealable microwave tube]Pyrimidine (210mg, 0.681mmol, Eq: 1.00), 3, 4-dimethylphenylboronic acid (112mg, 0.749mmol, Eq: 1.1) and potassium carbonate (376mg, 2.72mmol, Eq: 4.00) with DME (4 mL). Adding Pd (PPh)₃)₄(79mg, 0.0681mmol, Eq: 0.1). The reaction mixture was heated in a microwave at 160 ℃ for 60 minutes. The resulting solution was diluted with EtOAc and washed with brine. The combined organic phases were dried over anhydrous sodium sulfate and the solvent was removed under reduced pressure. The crude material was purified by column chromatography (silica gel, 5-70% ethyl acetate in hexane). The title compound was obtained as a solid (17mg, 11% yield). LC/MS: calculated m/z value C₁₄H₁₃N₃([M+H]⁺): 224.2, measurement: 224.2.

examples I to 7

4-p-tolyl-7H-pyrrolo [2,3-d ] pyrimidine

4-chloro-7-tosyl-7H-pyrrolo [2,3-d ] combined in 2mL of water in a 10mL sealable microwave tube]Pyrimidine (210mg, 0.681mmol, Eq: 1.00), p-tolylboronic acid (102mg, 0.749mmol, Eq: 1.1) and potassium carbonate (376mg, 2.72mmol, Eq: 4.00) with DME (4 mL). Adding Pd (PPh)₃)₄(79mg, 0.0681mmol, Eq: 0.1). The reaction mixture was heated in a microwave at 160 ℃ for 60 minutes. The resulting solution was diluted with EtOAc and washed with brine. The combined organic phases were dried over anhydrous sodium sulfate and the solvent was removed under reduced pressure. The crude material was purified by column chromatography (silica gel, 5-70% ethyl acetate in hexane to give the title compound as a solid (60mg, 42% yield.) LC/MS calcd for C by m/z₁₃H₁₁N₃([M+H]⁺): 210.2, measurement: 210.2.

examples I to 8

4- (3-chloro-4-methyl-phenyl) -7H-pyrrolo [2,3-d ] pyrimidine

4-chloro-7-tosyl-7H-pyrrolo [2,3-d ] combined in 2mL of water in a 10mL sealable microwave tube]Pyrimidine (210mg, 0.681mmol, Eq: 1.00), 3-chloro-4-methylphenylboronic acid (128mg, 0.749mmol, Eq: 1.1) and potassium carbonate (376mg, 2.72mmol, Eq: 4.00) with DME (4 mL). Adding Pd (PPh)₃)₄(79mg, 0.068mmol, Eq: 0.1). The reaction mixture was heated in a microwave at 160 ℃ for 60 minutes. The resulting solution was diluted with EtOAcAnd washed with brine. The combined organic phases were dried over anhydrous sodium sulfate and the solvent was removed under reduced pressure. The crude material was triturated with DCM. The solid was filtered. The title compound was obtained as a solid (14mg, 8% yield). LC/MS: calculated m/z value C₁₃H₁₀ClN₃([M+H]⁺): 244.7, measurement: 244.2.

examples I to 9

4-tert-butyl-N- [4- (7H-pyrrolo [2,3-d ] pyrimidin-4-yl) -benzyl ] -benzamide

Step 1: [4- (7H-pyrrolo [2,3-d ] pyrimidin-4-yl) -benzyl ] -carbamic acid tert-butyl ester

4-chloro-7H-pyrrolo [2,3-d ] was combined in a 20mL sealable microwave tube]Pyrimidine (500mg, 3.26mmol, Eq: 1.00), tert-butyl 4- (4, 4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) benzylcarbamate (1.6g, 4.8mmol, Eq: 1.47) and potassium carbonate (1.8g, 13.0mmol, Eq: 4.00) with DME (10mL) and water (5 mL). Adding Pd (PPh)₃)₄(376mg, 0.326mmol, Eq: 0.1), the reaction mixture was heated at 150 ℃ for 60 minutes. The resulting solution was diluted with EtOAc and washed with brine. The combined organic phases were dried over anhydrous sodium sulfate and the solvent was removed under reduced pressure. The crude material was purified by column chromatography (silica gel, 5-70% ethyl acetate in hexane). The title compound was obtained as a solid (545mg, 52% yield). LC/MS: calculated m/z value C₁₈H₂₀N₄O₂([M+H]⁺): 325.3, measurement: 325.2.

step 2: 4-tert-butyl-N- [4- (7H-pyrrolo [2,3-d ] pyrimidin-4-yl) -benzyl ] -benzamide

In a 20mL scintillation vial, 4- (7H-pyrrolo [2,3-d ] is dissolved with 2mL DCM and 2mL TFA]Pyrimidin-4-yl) benzylcarbamic acid tert-butyl ester (200mg, 0.617mmol, Eq: 1.00). The reaction mixture was stirred at room temperature for 30 minutes. The solvent was removed under reduced pressure. The crude material was dissolved in DMF (4 mL). 4-tert-butylbenzoic acid (121mg, 0.678mmol, Eq: 1.1), DIPEA (0.43mL, 2.47mmol, Eq: 4.00), and HATU (258mg, 0.678mmol, Eq: 1.1) were added. The reaction mixture was stirred at room temperature overnight. The reaction mixture was diluted with EtOAc and water and then stirred at rt for 30 min. The resulting solution was washed with brine. The combined organic phases were dried over anhydrous sodium sulfate and the solvent was removed under reduced pressure. The crude material was triturated with DCM and the resulting solid filtered to give the title compound as a solid (65mg, 27% yield). LC/MS: calculated m/z value C₂₄H₂₄N₄O([M+H]⁺): 385.4, measurement: 385.1.

examples I to 10

3-chloro-N- [4- (7H-pyrrolo [2,3-d ] pyrimidin-4-yl) -benzyl ] -benzamide

In a similar manner to example 9, step 2, using 3-chlorobenzoic acid (96.5mg, 0.617mmol, Eq: 1.00), the title compound was obtained as a solid (45mg, 20% yield). LC/MS: calculated m/z value C₂₀H₁₅ClN₄O([M+H]⁺): 363.8, measurement: 363.0.

examples I to 11

2- (3-chloro-phenylamino) -N- [4- (7H-pyrrolo [2,3-d ] pyrimidin-4-yl) -benzyl ] -acetamide

Step 1: (3-chloro-phenylamino) -acetic acid methyl ester

In a 250mL round bottom flask, methyl bromoacetate (1.66g, 1mL, 10.9mmol, Eq: 1.00), 3-chloroaniline (1.66g, 1.4mL, 13.0mmol, Eq: 1.2) and DIPEA (1.9mL, 10.9mmol, Eq: 1.00) were combined with DMF (20mL) to give a light yellow solution. The reaction mixture was heated at 60 ℃ overnight. The reaction mixture was diluted with EtOAc and then washed with brine. The combined organic phases were dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure to give the title compound (2.1g, 97% yield). LC/MS: calculated m/z value C₉H₁₀ClNO₂([M+H]⁺): 200.6, measurement: 200.0.

step 2: (3-chloro-phenylamino) -acetic acid

In a 20mL scintillation vial, in 5mL H₂Combine (3-chloro-phenylamino) -acetic acid methyl ester (500mg, 2.5mmol, Eq: 1.00) and NaOH (500mg, 12.5mmol, Eq: 4.99) with EtOH (8mL) in O to give a light yellow solution. The reaction mixture was heated at 60 ℃ for 4 hours. The reaction mixture was diluted with EtOAc and washed with 10% aqueous HCl. The combined organic phases were dried over anhydrous sodium sulfate and then evaporated under reduced pressure to give the title compound as a brown solid. LC/MS: calculated m/z value C₈H₈ClNO₂([M+H]⁺): 186.0, measurement: 186.0.

and step 3: 2- (3-chloro-phenylamino) -N- [4- (7H-pyrrolo [2,3-d ] pyrimidin-4-yl) -benzyl ] -acetamide

In analogy to example 9, step 2, using 4- (7H-pyrrolo [2, 3-d)]Pyrimidin-4-yl) benzylcarbamic acid tert-butyl ester (345mg, 1.06mmol, Eq: 1.00), 2- (3-chlorophenylamino) acetic acid (217mg, 1.17mmol, Eq: 1.1), DIPEA (740mg, 1mL, 5.73mmol, Eq: 5.38) and HATU (445mg, 1.17mmol, Eq: 1.1) to yield the title compound as a solid (71mg, 17% yield). LC/MS: calculated m/z value C₂₁H₁₈ClN₅O([M+H]⁺): 392.8, measurement: 392.1.

examples I to 12

4-tert-butyl-N- [ 2-fluoro-4- (7H-pyrrolo [2,3-d ] pyrimidin-4-yl) -benzyl ] -benzamide

Step 1: [ 2-fluoro-4- (4, 4,5, 5-tetramethyl- [1, 3, 2] dioxaborolan-2-yl) -benzyl ] -carbamic acid tert-butyl ester

In a pressure tube, combine tert-butyl 4-bromo-2-fluorobenzylcarbamate (5g, 16.4mmol), bis (pinacolato) diboron (6.26g, 24.7mmol) and potassium acetate (4.84g, 49.3mmol) with NMP (75.0mL) to give a light yellow solution. Degassing the reaction mixture in a nitrogen atmosphereFor 10 minutes. Adding [1, 1' -bis (diphenylphosphino) ferrocene]Dichloro-palladium (II) (722mg, 0.986 mmol). The reaction mixture was heated at 100 ℃ for 20 hours. The reaction mixture was quenched with water and extracted with DCM (3 × 100 mL). The combined organic layers were washed with water, brine, and Na₂SO₄Drying, filtering and concentrating. The crude material was purified by flash chromatography (silica gel, 120g, 0% -30% ethyl acetate in hexane). To obtain [ 2-fluoro-4- (4, 4,5, 5-tetramethyl- [1, 3, 2)]Dioxolane-2-yl-benzyl]Tert-butyl carbamate (5.8g, 100%) as yellow oil.

Step 2: [ 2-fluoro-4- (7H-pyrrolo [2,3-d ] pyrimidin-4-yl) -benzyl ] -carbamic acid tert-butyl ester

4-chloro-7-tosyl-7H-pyrrolo [2,3-d ] combined in 2mL of water in a 10mL sealable microwave tube]Pyrimidine (157mg, 0.511mmol, Eq: 1.00), [ 2-fluoro-4- (4, 4,5, 5-tetramethyl- [1, 3, 2-methyl ]]Dioxolane-2-yl-benzyl]Tert-butyl carbamate (197mg, 0.562mmol, Eq: 1.1) and potassium carbonate (282mg, 2.04mmol, Eq: 4.00) with DME (4 mL). Adding Pd (Ph)₃P)₄(59.0mg, 0.051mmol, Eq: 0.1), the reaction mixture was heated at 160 ℃ for 60 minutes. The resulting solution was diluted with EtOAc and washed with brine. The combined organic phases were dried over anhydrous sodium sulfate and the solvent was removed under reduced pressure. The crude material was purified by column chromatography (silica gel, 5-70% ethyl acetate in hexane). The title compound was obtained as a solid (75mg, 43% yield). LC/MS: calculated m/z value C₁₈H₁₉FN₄O₂([M+H]⁺): 343.3, measurement: 343.3.

and step 3: 4-tert-butyl-N- [ 2-fluoro-4- (7H-pyrrolo [2,3-d ] pyrimidin-4-yl) -benzyl ] -benzamide

In a 20mL scintillation vial, [ 2-fluoro-4- (7H-pyrrolo [2,3-d ] is dissolved with 2mL DCM and 2mL TFA]Pyrimidine 4-yl) -benzyl]-carbamic acid tert-butyl ester (70mg, 0.134mmol, Eq: 1.00). The reaction mixture was stirred at room temperature for 30 minutes. The solvent was removed under reduced pressure. The crude material was dissolved in DMF (4 mL). 4-tert-butylbenzoic acid (26mg, 0.148mmol, Eq: 1.1), DIPEA (0.047mL, 0.269mmol, Eq: 4.00) and HATU (56mg, 0.148mmol, Eq: 1.1) were added. The reaction mixture was stirred at room temperature overnight. The reaction mixture was diluted with EtOAc and water and then stirred at rt for 30 min. The resulting solution was washed with brine. The combined organic phases were dried over anhydrous sodium sulfate and the solvent was removed under reduced pressure. The crude material was triturated with DCM and the resulting solid filtered to give the title compound as a solid (32mg, 59% yield). LC/MS: calculated m/z value C₂₄H₂₃FN₄O([M+H]⁺): 403.4, measurement: 403.2.

examples I to 13

4- {4- [ (4-tert-butyl-benzoylamino) -methyl ] -3-fluoro-phenyl } -7H-pyrrolo [2,3-d ] pyrimidine-6-carboxylic acid tert-butyl ester

Step 1: 4-chloro-5-hydroxy-6, 7-dihydro-5H-pyrrolo [2,3-d ] pyrimidine-6-carboxylic acid tert-butyl ester

To a suspension of 4, 6-dichloropyrimidine-5-carbaldehyde (2g, 11.3mmol, Eq: 1.00) in EtOH (50mL) was added tert-butyl 2-aminoacetate (1.48g, 11.3mmol, Eq: 1.00), followed by triethylamine (2.86g, 3.94mL, 28.3mmol, Eq:2.5), stirring at r.t. for 48 h. The solvent was removed under reduced pressure. The crude material was diluted with dichloromethane and washed with water. The combined organic phases were dried over anhydrous sodium sulfate and the solvent was removed under reduced pressure. The crude material was purified by column chromatography (silica gel, 10-90% ethyl acetate in hexanes) to give the title compound (634mg, 21% yield) as a white solid. LC/MS: calculated m/z value C₁₁H₁₄ClN₃O([M+H]⁺): 272.7, measurement: 272.1.

step 2: 4-chloro-7H-pyrrolo [2,3-d ] pyrimidine-6-carboxylic acid tert-butyl ester

To 4-chloro-5-hydroxy-6, 7-dihydro-5H-pyrrolo [2,3-d ] at 0 DEG C]To a solution of tert-butyl pyrimidine-6-carboxylate (634mg, 2.33mmol, Eq: 1.00) in DMF (10mL) was added sodium hydride (93.3mg, 2.33mmol, Eq: 1.00), followed by stirring at r.t. for 1 h. The reaction is stopped with water and then with NH₄Cl and brine. The combined organic layers were dried over anhydrous sodium sulfate, and then the solvent was removed under reduced pressure. The crude material was purified by column chromatography (silica gel, 5-35% ethyl acetate in hexane) to give the title compound (417mg, 70.4% yield) as a white solid. LC/MS: calculated m/z value C₁₁H₁₂ClN₃O([M+H]⁺): 254.6, measurement: 254.1.

and step 3: n- (4-bromo-2-fluoro-benzyl) -4-tert-butyl-benzamide

To a solution of (4-bromo-2-fluorophenyl) methylamine (1.5g, 7.35mmol, Eq: 1.00) cooled to 0 ℃ in DCM (25mL) was added 4-tert-butylbenzoyl chloride (1.45g, 7.35mmol, Eq: 1.00), triethylamine 1.49g, 2.05mL, 14.7mmol, Eq: 2.00) solution in DCM (5 mL). The reaction mixture was warmed to r.t.1hr. By passingThe reaction mixture was purified by column chromatography (silica gel, 5-40% ethyl acetate in hexane) to give the title compound (2.59g, 7.11mmol, 96.7% yield) as a white solid.¹H NMR (400MHz, chloroform-d) ppm 7.74(d, J ═ 8.0Hz, 2H), 7.48(d, J ═ 8.5Hz, 2H), 7.35(t, J ═ 7.5Hz, 1H), 6.48(s, 1H), 4.67(d, J ═ 5.7Hz, 2H), 1.36(s, 9H).

And 4, step 4: 4-tert-butyl-N- [ 2-fluoro-4- (4, 4,5, 5-tetramethyl- [1, 3, 2] dioxaborolan-2-yl) -benzyl ] -benzamide

Under stirring in N₂N- (4-bromo-2-fluorobenzyl) -4-tert-butylbenzamide (600mg, 1.65mmol, Eq: 1.00), 4, 4, 4 ', 4 ', 5, 5, 5 ', 5 ' -octamethyl-2, 2 ' -bis (1, 3, 2-dioxaborolan) (627mg, 2.47mmol, Eq: 1.5), potassium acetate (485mg, 4.94mmol, Eq: 3) and PdCl in an atmosphere₂(dppf)-CH₂Cl₂NMP (12mL) was added to a mixture of (121mg, 165. mu. mol, Eq: 0.1) and heated to 100 ℃ for 16 h. The reaction mixture was diluted with EtOAc, washed with water and brine. The combined organic layers were dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure. The crude material was purified by column chromatography (silica gel, 15-60% ethyl acetate in hexanes) to give the title compound (545mg, 80% yield) as an off-white solid.¹H NMR (400MHz, chloroform-d) ppm 7.74(d, J ═ 8.3Hz, 2H), 7.75(d, J ═ 7.4Hz, 1H), 7.51(d, J ═ 10.4Hz, 1H), 7.47(d, J ═ 8.4Hz, 2H), 7.44(t, J ═ 7.0Hz, 1H), 6.46(s, 1H), 4.74(d, J ═ 5.9Hz, 2H), 1.37(s, 12H), 1.36(s, 9H).

And 5: 4- {4- [ (4-tert-butyl-benzoylamino) -methyl ] -3-fluoro-phenyl } -7H-pyrrolo [2,3-d ] pyrimidine-6-carboxylic acid tert-butyl ester

To 4-chloro-7H-pyrrolo [2,3-d]Tert-butyl pyrimidine-6-carboxylate (100mg, 0.394mmol, Eq: 1.00), 4-tert-butyl-N- (2-fluoro-4- (4, 4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) benzyl) benzamide (162mg, 0.394mmol, Eq: 1.00), tetrakis (triphenylphosphine) palladium (0) (46mg, 0.04mmol, Eq: 0.1) and potassium carbonate (163mg, 1.18mmol, Eq: 3.00) were added DME (1mL) and water (500. mu.L) to a mixture and heated in a microwave at 150 ℃ for 30 min. The reaction mixture was filtered through a pad of celite and diluted with dichloromethane. The solution was purified by column chromatography (silica gel, 15-60% ethyl acetate in hexane, then 0-30% [ 10% methanol/dichloromethane ]]Dichloromethane solution) to yield the title compound (80mg, 40% yield) as an off-white solid.¹H NMR (400MHz, chloroform-d) ppm 10.93(s, 1H), 9.18(s, 1H), 7.94(t, J ═ 9.3Hz, 2H), 7.80(d, J ═ 8.5Hz, 2H), 7.68(t, J ═ 7.8Hz, 1H), 7.50(d, J ═ 8.4Hz, 1H), 7.39(s, 1H), 6.67(t, J ═ 5.9Hz, 1H), 4.83(d, J ═ 5.8Hz, 2H), 1.69(s, 9H), 1.37(s, 9H); LC/MS: calculated m/z value C₂₉H₃₁FN₄O₃([M+H]⁺): 503.5, measurement: 503.3.

examples I to 14

4- {4- [ (4-tert-butyl-benzoylamino) -methyl ] -3-fluoro-phenyl } -7H-pyrrolo [2,3-d ] pyrimidine-6-carboxylic acid

To 4- {4- [ (4-tert-butyl-benzoylamino) -methyl]-3-fluoro-phenyl } -7H-pyrrolo [2,3-d]To a solution of pyrimidine-6-carboxylic acid tert-butyl ester (55mg, 0.109mmol, Eq: 1.00) in dichloromethane (1mL) was added trifluoroacetic acid (843. mu.L, 10.9mmol, Eq: 100) and stirred at r.t. for 2 h. Concentration in vacuo removed solvent (3X) from methanol to give the title compound (43mg, 88%Yield) as a light brown solid.¹H NMR(400MHz，DMSO-d₆) ppm 13.05(s, 1H), 9.10(t, J ═ 5.9Hz, 1H), 8.98(s, 1H), 8.06(d, J ═ 8.2Hz, 1H), 7.97(d, J ═ 11.2Hz, 1H), 7.89(d, J ═ 8.4Hz, 2H), 7.59(t, J ═ 7.6Hz, 1H), 7.54(m, 3H), 4.64(d, J ═ 5.6Hz, 2H), 1.33(s, 9H); LC/MS: calculated m/z value C₂₅H₂₃FN₄O₃([M+H]⁺): 447.4, measurement: 447.2.

examples I to 15

4-tert-butyl-N- (2-fluoro-4- (6- (morpholine-4-carbonyl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl) benzyl) benzamide

To 4- {4- [ (4-tert-butyl-benzoylamino) -methyl]-3-fluoro-phenyl } -7H-pyrrolo [2,3-d]To a solution of pyrimidine-6-carboxylic acid (50mg, 0.112mmol, Eq: 1.00), HBTU (42.5mg, 0.112mmol, Eq: 1.00) and DIPEA (59. mu.l, 0.336mmol, Eq: 3) in DMF (1.5mL) was added morpholine (19.5mg, 0.224mmol, Eq: 2.00), and the mixture was stirred at room temperature for 16 h. The reaction mixture was diluted with ethyl acetate and washed with brine. The combined organic phases were dried over anhydrous sodium sulfate and the solvent was removed under reduced pressure. The crude material was purified by column chromatography (silica gel, 0-5% methanol in dichloromethane, then 50-100% ethyl acetate in hexanes) to give the title compound (26mg, 45% yield) as a white solid.¹H NMR (400MHz, chloroform-d) ppm 8.96(s, 1H), 7.74(m, 2H), 7.71(d, J ═ 8.4Hz, 2H), 7.58(t, J ═ 8.2Hz, 1H), 7.41(d, J ═ 8.4Hz, 1H), 6.91(s, 1H), 4.71(s, 1H), 3.81(br.s, 4H), 3.72(br.s, 4H), 1.27(s, 9H).

Examples I to 16

4- {4- [ (4-tert-butyl-benzoylamino) -methyl ] -3-fluoro-phenyl } -7H-pyrrolo [2,3-d ] pyrimidine-6-carboxylic acid dimethylamide

To 4- {4- [ (4-tert-butyl-benzoylamino) -methyl]-3-fluoro-phenyl } -7H-pyrrolo [2,3-d]To a solution of pyrimidine-6-carboxylic acid (50mg, 0.112mmol, Eq: 1.00), HBTU (42.5mg, 0.112mmol, Eq: 1.00) and DIPEA (59. mu.l, 0.336mmol, Eq: 3.00) in DMF (1mL) was added a THF solution of dimethylamine (112. mu.l, 0.224mmol, Eq: 2.00) and stirred at room temperature for 16 h. The reaction mixture was diluted with ethyl acetate and washed with brine. The combined organic phases were dried over anhydrous sodium sulfate and the solvent was removed under reduced pressure. The crude material was purified by column chromatography (silica gel, gradient 50-100% ethyl acetate in hexanes) to give the title compound (18mg, 34% yield) as a white solid.¹H NMR (400MHz, DMSO-d) ppm 12.78(s, 1H), 9.10(t, J ═ 5.8Hz, 1H), 8.92(s, 1H), 8.04(d, J ═ 8.0Hz, 1H), 7.96(d, J ═ 11.6Hz, 1H), 7.89(d, J ═ 8.6Hz, 2H), 7.56(t, J ═ 7.9Hz, 1H), 7.53(d, J ═ 8.3Hz, 2H), 7.23(s, 1H), 4.62(s, 2H), 3.26(br.s, 3H), 3.06(br.s, 3H), 1.32(s, 9H); LC/MS: calculated m/z value C₂₇H₂₈FN₅O₂([M+H]⁺): 474.5, measurement: 474.3.

examples I to 17

4- {4- [ (4-tert-butyl-benzoylamino) -methyl ] -3-fluoro-phenyl } -7H-pyrrolo [2,3-d ] -6-carboxylic acid methylamide

To 4- {4- [ (4-tert-butyl-benzoylamino) -methyl]-3-fluoro-phenyl } -7H-pyrrolo [2,3-d]Pyrimidine-6-carboxylic acid (50 mg)0.112mmol, Eq: 1.00), HBTU (42.5mg, 0.112mmol, Eq: 1.00) and DIPEA (43mg, 59 μ l, 0.336mmol, Eq: 3.00) solution in DMF (1.00mL) was added methylamine in THF (112 μ l, 0.224mmol, Eq: 2.00), stirred at room temperature for 16 h. The reaction mixture was diluted with ethyl acetate and washed with brine. The combined organic phases were dried over anhydrous sodium sulfate and the solvent was removed under reduced pressure. The crude material was purified by column chromatography (silica gel, 50-100% ethyl acetate in hexanes) to give the title compound (25mg, 48.6% yield) as a white solid.¹H NMR(400MHz，DMSO-d₆) ppm 12.82(s, 1H), 9.11(t, J ═ 5.7Hz, 1H), 8.92(s, 1H), 8.70(d, J ═ 4.8Hz, 1H), 8.01(d, J ═ 7.8Hz, 1H), 7.93(d, J ═ 11.6Hz, 1H), 7.89(d, J ═ 8.7Hz, 2H), 7.61(s, 1H), 7.59(t, J ═ 8.1Hz, 1H), 7.53(d, J ═ 8.4Hz, 2H), 4.64(d, J ═ 5.4Hz, 2H), 2.84(d, J ═ 4.5Hz, 3H), 1.32(s, 9H); LC/MS: calculated m/z value C₂₆H₂₆FN₅O₂([M+H]⁺): 460.5, measurement: 460.3.

examples I to 18

4- {4- [ (4-tert-butyl-benzoylamino) -methyl ] -3-fluoro-phenyl } -7H-pyrrolo [2,3-d ] pyrimidine-6-carboxylic acid (2-hydroxy-ethyl) -amide

To 4- {4- [ (4-tert-butyl-benzoylamino) -methyl]-3-fluoro-phenyl } -7H-pyrrolo [2,3-d]To a solution of pyrimidine-6-carboxylic acid (50mg, 0.112mmol, Eq: 1.00), HBTU (42.5mg, 0.112mmol, Eq: 1.00) and DIPEA (59. mu.l, 0.336mmol, Eq: 3.00) in DMF (1.00mL) was added 2-aminoethanol (14mg, 0.224mmol, Eq: 2.00), and the mixture was stirred at room temperature for 16 h. The reaction mixture was diluted with ethyl acetate and washed with brine. The combined organic phases were dried over anhydrous sodium sulfate and the solvent was removed under reduced pressure. The crude material was purified by column chromatography (silica gel, 50-100% ethyl acetate)Hexane solution of ethyl acetate, then containing NH₄OH in 0-10% methanol in dichloromethane) to give the title compound (16mg, 29% yield) as an off-white solid.¹H NMR(400MHz，DMSO-d₆) ppm 12.80(s, 1H), 9.10(t, J ═ 5.6Hz, 1H), 8.92(s, 1H), 8.73(t, J ═ 6.4Hz, 1H), 8.03(d, J ═ 8.6Hz, 1H), 7.95(d, J ═ 11.0Hz, 1H), 7.89(d, J ═ 8.4Hz, 2H), 7.69(s, 1H), 7.59(t, J ═ 7.5Hz, 1H), 7.53(d, J ═ 8.2Hz, 2H), 4.64(d, J ═ 6.0Hz, 2H), 3.55(t, J ═ 5.2Hz, 2H), 3.38(m, 2H), 1.32(s, 9H); LC/MS: calculated m/z value C₂₇H₂₈FN₅O₃([M+H]⁺): 490.5, measurement: 490.4.

examples I to 19

4- {4- [ (4-tert-butyl-benzoylamino) -methyl ] -3-fluoro-phenyl } -7H-pyrrolo [2,3-d ] pyrimidine-6-carboxylic acid (2-dimethylamino-ethyl) -amide

To the cooled to 0 deg.C 4- {4- [ (4-tert-butyl-benzoylamino) -methyl]-3-fluoro-phenyl } -7H-pyrrolo [2,3-d]Pyrimidine-6-carboxylic acid (50mg, 0.112mmol, Eq: 1.00), N¹，N¹A solution of dimethylethylene-1, 2-diamine (10mg, 0.112mmol, Eq: 1.00) and DIPEA (49. mu.l, 0.280mmol, Eq: 2.5) in DMF (1.00mL) was added 1-propanephosphonic cyclic anhydride (80. mu.l, 0.134mmol, Eq: 1.2) and warmed to r.t.4h. The reaction mixture was diluted with ethyl acetate and washed with brine. The combined organic phases were dried over anhydrous sodium sulfate and the solvent was removed under reduced pressure. Purification of the crude material (containing NH) by column chromatography₄OH in 0-15% methanol in dichloromethane) to give the title compound (25mg, 43% yield) as an off-white solid.¹H NMR(400MHz，METHANOL-d₄)ppm8.93(s，1H)，7.97(d，J＝8.1Hz，1H)，7.91(d，J＝11.0Hz，1H)，7.86(d，J＝8.5Hz, 2H), 7.66(t, J ═ 7.8Hz, 1H), 7.57(s, 1H), 7.56(d, J ═ 8.5Hz, 2H), 4.76(s, 2H), 3.75(t, J ═ 6.1Hz, 2H), 3.22(m, 2H), 2.84(s, 6H), 1.38(s, 9H); LC/MS: calculated m/z value C₂₉H₃₃FN₆O₂([M+H]⁺): 517.6, measurement: 517.4.

examples I to 20

4-tert-butyl-N- {1- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -piperidin-4-ylmethyl } -benzamide

Step 1: 4- [ (4-tert-butyl-benzoylamino) -methyl ] -piperidine-1-carboxylic acid tert-butyl ester

To a solution of tert-butyl 4- (aminomethyl) piperidine-1-carboxylate (1g, 4.67mmol, Eq: 1.00) in DCM (16.7mL) cooled to 0 deg.C was added a solution of 4-tert-butylbenzoyl chloride (918mg, 4.67mmol, Eq: 1.00), triethylamine (361. mu.l, 4.67mmol, Eq: 1.00) in DCM (5 mL). The reaction mixture was warmed to r.t.1h. The reaction mixture was purified by column chromatography (30-70% ethyl acetate in hexane) to give the title compound (1.75g, 4.67mmol, 100% yield) as a colorless viscous oil.¹H NMR (400MHz, chloroform-d) ppm 7.73(d, J ═ 8.5Hz, 2H), 7.48(d, J ═ 8.1Hz, 2H), 6.21(s, 1H), 4.15(d, J ═ 12.9Hz, 2H), 3.39(t, J ═ 6.0Hz, 2H), 2.72(t, J ═ 14.7Hz, 2H), 1.83(m, 1H), 1.76(d, J ═ 14.3Hz, 2H), 1.48(s, 9H), 1.36(s, 9H).

Step 2: 4-tert-butyl-N-piperidin-4-ylmethyl-benzamides

To 4- [ (4-tert-butyl-benzoylamino) -methyl]To a solution of tert-butyl-piperidine-1-carboxylate (1.75g, 4.67mmol, Eq: 1.00) in DCM (35mL) was added trifluoroacetic acid (7.2mL, 93.5mmol, Eq: 20) and stirred at r.t. for 4 h. The solvent was removed under reduced pressure and dried in vacuo to give the title compound (2.92g, 124% yield) as a viscous colorless oil.¹H NMR (400MHz, chloroform-d) ppm 8.72(br.s, 3H), 7.71(d, J ═ 8.5Hz, 2H), 7.49(d, J ═ 8.5Hz, 2H), 6.81(t, J ═ 6.7Hz, 1H), 3.56(d, J ═ 12.8Hz, 2H), 3.46(t, J ═ 6.4Hz, 2H), 3.01(q, J ═ 11.7Hz, 2H), 2.02(d, J ═ 14.3Hz, 2H), 1.69(q, J ═ 14.1Hz, 2H), 1.36(s, 9H).

And step 3: n- [1- (7-benzenesulfonyl-6-iodo-7H-pyrrolo [2,3-d ] pyrimidin-4-yl) -piperidin-4-ylmethyl ] -4-tert-butyl-benzamide

To 4-chloro-6-iodo-7- (phenylsulfonyl) -7H-pyrrolo [2,3-d]To a suspension of pyrimidine (200mg, 0.477mmol, Eq: 1.00) in EtOH (3.00mL) was added 4-tert-butyl-N-piperidin-4-ylmethyl-benzamide (185mg, 477. mu. mol, Eq: 1.00) and triethylamine (332. mu.l, 2.38mmol, Eq: 5.00) and heated to 80 ℃ for 2 h. The reaction mixture was cooled to r.t. Precipitate formed and was filtered off. The filtrate (silica gel, 20-80% ethyl acetate in hexane) was purified by column chromatography to give the title compound (159mg, 51% yield) as a white solid.¹H NMR (400MHz, chloroform-d) ppm8.37(s, 1H), 8.25(d, J ═ 8.21Hz, 2H), 7.72(d, J ═ 8.4Hz, 2H), 7.63(t, J ═ 8.1Hz, 1H), 7.54(t, J ═ 7.6Hz, 2H), 7.47(d, J ═ 8.5Hz, 2H), 6.96(s, 1H), 6.28(s, 1H), 4.61(d, J ═ 12.4Hz, 2H), 3.40(t, J ═ 6.3Hz, 2H), 3.09(t, J ═ 13.3Hz, 2H), 2.03(m, 1H), 1.91(d,J＝13.3Hz，2H)，1.36(s，9H)。

and 4, step 4: n- {1- [ 7-benzenesulfonyl-6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -piperidin-4-ylmethyl } -4-tert-butyl-benzamide

To the N- [1- (7-benzenesulfonyl-6-iodo-7H-pyrrolo [2,3-d ]]Pyrimidin-4-yl) -piperidin-4-ylmethyl]-4-tert-butyl-benzamide (159mg, 0.242mmol, Eq: 1.00), 1-methyl-1H-pyrazol-4-ylboronic acid (36.5mg, 0.290mmol, Eq: 1.2), Pd (PPh)₃)₄To a mixture of (28mg, 0.024mmol, Eq: 0.1) and potassium carbonate (100mg, 0.725mmol, Eq: 3.00) was added DME (1.29 ml)/water (322. mu.l) and heated in a microwave to 150 ℃ for 1 h. The reaction mixture was diluted with DCM and washed with water. The combined organic phases were dried over anhydrous sodium sulfate and the solvent was removed under reduced pressure. The crude material was purified by column chromatography (silica gel, 30-100% ethyl acetate in hexanes) to give the title compound (85mg, 58% yield) as a white solid. LC/MS: calculated m/z value C₃₃H₃₇N₇O₃S([M+H]⁺): 612.7, measurement: 612.4.

and 5: 4-tert-butyl-N- {1- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -piperidin-4-ylmethyl } -benzamide

To the N- {1- [ 7-benzenesulfonyl-6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d]Pyrimidin-4-yl]-piperidin-4-ylmethyl } -4-tert-butyl-benzamide (85mg, 0.139mmol, Eq: 1.00) in THF (926. mu.l)/MeOH (463. mu.l) cesium carbonate (136mg, 0.417mmol, Eq: 3.00) was added and stirred at r.t. for 16 h. The reaction mixture was purified by column chromatography (silica gel, containing NH)₄1-6 of OH% methanol in DCM) to give the title compound (45mg, 68.7% yield) as a white solid.¹H NMR(400MHz，DMSO-d₆) ppm 11.95(s, 1H), 8.45(t, J ═ 5.4Hz, 1H), 8.11(s, 1H), 7.93(s, 1H), 7.80(d, J ═ 7.9Hz, 2H), 7.49(d, J ═ 7.9Hz, 2H), 6.75(s, 1H), 4.71(d, J ═ 13.8Hz, 2H), 3.88(s, 3H), 3.20(t, J ═ 6.2Hz, 2H), 3.06(t, J ═ 12.8Hz, 2H), 1.97(m, 1H), 1.82(d, J ═ 13.3Hz, 2H), 1.31(s, 9H), 1.23(d, J ═ 14.3Hz, 2H); LC/MS: calculated m/z value C₂₇H₃₃N₇O([M+H]⁺): 472.6, measurement: 472.4.

examples I to 21

4-tert-butyl-N- {4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzyl } -benzamide

Step 1: 4- [7- (toluene-4-sulfonyl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzyl } -carbamic acid tert-butyl ester

4-chloro-7-tosyl-7H-pyrrolo [2,3-d ] combined in 5mL of water in a 20mL sealable microwave tube]Pyrimidine (1g, 3.25mmol, Eq: 1.00), 4- ((tert-butoxycarbonylamino) methyl) phenylboronic acid (1.22g, 4.87mmol, Eq: 1.5) and potassium carbonate (1.8g, 13.0mmol, Eq: 4.00) with DME (10 mL). Adding Pd (PPh)₃)₄(375mg, 0.325mmol, Eq: 0.1). The reaction mixture was heated in a microwave at 150 ℃ for 30 minutes. The solution was washed with EtOAc and brine. The combined organic phases were dried over anhydrous sodium sulfate and the solvent was removed under reduced pressure. The crude material was purified by column chromatography (silica gel, 5-70% ethyl acetate)Hexane solution of). The title compound was obtained as a white solid (700mg, 45% yield). LC/MS: calculated m/z value C₂₅H₂₆N₄O₄S([M+H]⁺): 479.5, measurement: 479.3.

step 2: 4- [ 6-bromo-7- (toluene-4-sulfonyl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzyl } -carbamic acid tert-butyl ester

In a 100mL round-bottom flask, 4- [7- (toluene-4-sulfonyl) -7H-pyrrolo [2,3-d ] was placed]Pyrimidin-4-yl]-benzyl } -carbamic acid tert-butyl ester (500mg, 1.04mmol, Eq: 1.00) was dissolved in THF (10mL) and cooled to-78 ℃. LDA 2M solution in heptane/THF/ethylbenzene (1.31mL, 2.61mmol, Eq: 2.5) was added under nitrogen at-78C to give a dark brown solution. The reaction mixture was stirred at-78 ℃ for 1hr 30 min. 1, 2-dibromo-1, 1,2, 2-tetrachloroethane (851mg, 2.61mmol, Eq: 2.5) in 5mL of THF was added and the reaction mixture was stirred at-78C for 2 hours. Water was added. The reaction mixture was diluted with EtOAc and washed with brine. The combined organic phases were dried over anhydrous sodium sulfate and then evaporated. The crude material was purified by column chromatography (silica gel, 5-70% ethyl acetate in hexane). The title compound was obtained as a white solid (450mg, 77% yield). LC/MS: calculated m/z value C₂₅H₂₅BrN₄O₄S([M+H]⁺): 558.4, measurement: 558.8.

and step 3: {4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzyl } -carbamic acid tert-butyl ester

4- [ 6-bromo-7- (toluene-4-sulfonyl) -7H-pyrrolo [2,3-d ] is combined in water (1mL) in a 10mL sealable microwave tube]Pyrimidin-4-yl]-benzyl } -carbamic acid tert-butyl ester (200mg, 0.359mmol, Eq: 1.00), 1-methyl-4- (4, 4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole (192mg, 0.923mmol, Eq: 2.57) and potassium carbonate (198mg, 1.44mmol, Eq: 4.00) with DME (4 mL). Adding Pd (PPh)₃)₄(42mg, 0.036mmol, Eq: 0.1), the reaction mixture was sealed and heated in a microwave at 150 ℃ for 60 minutes. The reaction mixture was diluted with EtOAc and then washed with brine. The combined organic phases were dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure. The crude material was triturated with DCM then filtered to give the title compound as a yellow solid (32mg, 22% yield). LC/MS: calculated m/z value C₂₂H₂₄N₆O₂([M+H]⁺): 405.4, measurement: 405.2.

and 4, step 4: 4-tert-butyl-N- {4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzyl } -benzamide

Combine {4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d in a 20mL scintillation vial]Pyrimidin-4-yl]-benzyl } -carbamic acid tert-butyl ester (30mg, 0.074mmol, Eq: 1.00) with 1mL DCM and 1mL TFA. The solution was stirred at room temperature for 1 hour. The solvent was removed under reduced pressure. The crude material was dissolved in DMF (2 mL). 4-tert-butylbenzoic acid (14.5mg, 0.082mmol, Eq: 1.1), DIPEA (0.052mL, 0.297mmol, Eq: 4.00), and HATU (31.0mg, 0.082mmol, Eq: 1.1) were added. The reaction mixture was stirred overnight. The resulting solution was diluted with 10mL of water and 5mL of EtOAc. The solution was stirred at room temperature for a further 30 minutes. The organic phase was extracted and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure. The resulting solid was triturated with DCM and the resulting solid was filtered. The title compound was obtained as a solid (11mg, 32% yield). LC/MS: calculated m/z value C₂₈H₂₈N₆O([M+H]⁺): 465.5, measurement: 465.2.

examples I to 22

4-cyclopropyl-N- {4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzyl } -benzamide

Following a similar procedure to example 21, step 4, using 4-cyclopropylbenzoic acid (12.0mg, 0.074mmol, Eq: 1.00), the title compound was obtained as a solid (16mg, 48% yield). LC/MS: calculated m/z value C₂₇H₂₄N₆O([M+H]⁺): 449.5, measurement: 449.2.

examples I to 23

4-isopropyl-N- {4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzyl } -benzamide

The title compound was obtained as a solid (5mg, 15% yield) by a method similar to example 21, step 4, using 4-isopropylbenzoic acid (13.4mg, 0.082mmol, Eq: 1.1). LC/MS: calculated m/z value C₂₇H₂₆N₆O([M+H]⁺): 451.5, measurement: 451.3.

examples I to 24

N- {4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzyl } -4-oxetan-3-yl-benzamide

Step 1: 4-Oxetan-3-yl-benzoic acid methyl ester

In a 20mL sealable microwave vial, 4- (methoxycarbonyl) phenylboronic acid (978mg, 5.44mmol, Eq: 2.0), trand-2-aminocyclohexanol hydrochloride (50mg, 0.326mmol, Eq: 0.12) and nickel iodide (102mg, 0.326mmol, Eq: 0.12) were combined with isopropanol (8mL) to give a white suspension. NaHMDS (997mg, 5.44mmol, Eq: 2.0) was added. The reaction mixture was back-filled with argon and stirred for 5 minutes. 3-iodooxetane (0.5g, 2.72mmol, Eq: 1.00) was added. The reaction mixture was sealed and then heated in a microwave at 80 ℃ for 20 minutes. After this time, TLC showed 2 possible product spots, with Rf close to each other. The reaction mixture was diluted with isopropanol and then filtered through filter paper. The solvent was concentrated to give a yellow oil. The product was dissolved in dichloromethane and the solution was concentrated on silica gel. The crude product supported on silica gel was passed through a 40g silica gel column. Flash chromatography was performed (5% ethyl acetate-hexanes to 10% ethyl acetate-hexanes). The title compound was isolated as an oil (118mg, 23% yield). The other by-product isolated corresponded to isopropyl 4-oxetan-3-yl-benzoate (77mg, 13% yield).

Step 2: 4-Oxetan-3-yl-benzoic acid

In a 100mL pear-shaped flask, methyl 4- (oxetan-3-yl) benzoate (118mg, 0.614mmol, Eq: 1.00) and lithium hydroxide monohydrate (40mg, 0.953mmol, Eq: 1.55) were combined with THF (2.5mL) to give a colorless solution. Water (2.5mL) was added. The reaction mixture was stirred at room temperature overnight. In the morning, TLC showed a small amount of starting material remaining. An additional 40mg of LiOH monohydrate was added and the reaction mixture was stirred at room temperature overnight. The solvent was removed under reduced pressure. The aqueous residue was then diluted with 10mL of water, and the reaction mixture was diluted with 20mL of 1: the solution was extracted with 1 hexane-ethyl acetate. The aqueous phase was then acidified with a few drops of 4N aqueous HCl to give a white suspension. The suspension was extracted with ethyl acetate. The organic extract was dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure. The title compound was obtained as a white solid (88mg, 80% yield). The product was used without further purification.

And step 3: n- {4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzyl } -4-oxetan-3-yl-benzamide

Combine {4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d in a 20mL scintillation vial]Pyrimidin-4-yl]-benzyl } -carbamic acid tert-butyl ester (40mg, 0.099mmol, Eq: 1.00) with 3mL DCM and 3mL TFA. The solution was stirred at room temperature for 1 hour. The solvent was removed under reduced pressure. The crude material was dissolved in DMF (4 mL). 4-Oxetan-3-yl-benzoic acid (18mg, 0.099mmol, Eq: 1.00), DIPEA (0.069mL, 0.396mmol, Eq: 4.00) and HATU (38mg, 0.099mmol, Eq: 1.00) were added. The reaction mixture was stirred overnight. The resulting solution was diluted with 10mL of water and 5mL of EtOAc. The solution was stirred at room temperature for a further 30 minutes. The organic phase was extracted and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure. The resulting solid was triturated with DCM and the resulting solid was filtered. The title compound was obtained as a solid (3mg, 7% yield). LC/MS: calculated m/z value C₂₇H₂₄N₆O₂([M+H]⁺): 465.5, measurement: 465.2.

examples I to 25

4- (3-methyl-oxetan-3-yl) -N- {4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzyl } -benzamide

Step 1: 4- (3-methyl-oxetan-3-yl) -benzoic acid

In a 250mL 3-neck flask, 3- (4-bromophenyl) -3-methyloxetane (1.05g, 4.62mmol) was combined with THF (35mL) to give a colorless solution. The solution was cooled to-78 ℃ in a dry ice-acetone bath. To the cold solution was added dropwise a 1.6M solution of nBuLi in hexane (3.32mL, 5.32 mmol). Further dropwise over a period of 10 minutes. The reaction mixture was stirred at-78 ℃ for 1 hour. Thereafter, carbon dioxide gas generated in the flask separately from the dry ice was added to the reaction mixture through the long needle. The reaction mixture quickly turned light yellow. Carbon dioxide was bubbled for an additional 20 minutes at low temperature. Thereafter, the reaction mixture was a white suspension. The reaction mixture was warmed to room temperature and then quenched slowly with water. The organic solvent was evaporated. The resulting mixture was extracted with a 1: 1 solution of ethyl acetate and hexane. The aqueous phase was then brought to an acidic pH by addition of 4N aqueous HCl. The resulting white suspension was vacuum filtered using a buchner funnel. The collected white solid was further dried on a vacuum funnel and then further dried in a vacuum oven to give 4- (3-methyl-oxetan-3-yl) -benzoic acid (456mg, 51%).¹H NMR(300MHz，DMSO-d₆)ppm 12.89(br.s.，1H)，7.93(d，J＝8.48Hz，2H)，7.36(d，J＝8.67Hz，2H)，4.81(d，J＝5.84Hz，2H)，4.56(d，J＝6.03Hz，2H)，1.64(s，3H)。

Step 2: 4- (3-methyl-oxetan-3-yl) -N- {4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzyl } -benzamide

Combine {4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d in a 20mL scintillation vial]Pyrimidin-4-yl]-benzyl } -carbamic acid tert-butyl ester (60mg, 0.148mmol, Eq: 1.00) with 3mL DCM and 3mL TFA. The solution was stirred at room temperature for 1 hour. The solvent was removed under reduced pressure. The crude material was dissolved in DMF (4 mL). 4-Oxetan-3-yl-benzoic acid (31mg, 0.163mmol, Eq: 1.1), DIPEA (0.104mL, 0.593mmol, Eq: 4.00) and HATU (62mg, 0.163mmol, Eq: 1.1) were added. The reaction mixture was stirred overnight. The resulting solution was diluted with 10mL of water and 5mL of EtOAc. The solution was stirred at room temperature for a further 30 minutes. The organic phase was extracted and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure. The resulting solid was triturated with DCM and the resulting solid was filtered. The title compound was obtained as a yellow solid (15mg, 21% yield). LC/MS: calculated m/z value C₂₈H₂₆N₆O₂([M+H]⁺): 479.5, measurement: 479.2.

examples I to 26

4,5,6, 7-tetrahydro-benzo [ b ] thiophene-2-carboxylic acid 4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzylamide

Combine {4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d in a 20mL scintillation vial]Pyrimidin-4-yl]-benzyl } -carbamic acid tert-butyl ester (40mg, 0.099mmol, Eq: 1.00) with 3mL DCM and 3mL TFA. The solution was stirred at room temperature for 1 hour. The solvent was removed under reduced pressure. The crude material was dissolved in DMF (4 mL). Adding 4,5,6, 7-tetrahydro-benzo [ b ]]Thiophene-2-carboxylic acid (20mg, 0.109mmol, Eq: 1.1), DIPEA (0.069mL, 0.396mmol, Eq: 4.00), and HATU (41mg, 0.109mmol, Eq: 1.1). Mixing the reactionThe mixture was stirred overnight. The resulting solution was diluted with 10mL of water and 5mL of EtOAc. The solution was stirred at room temperature for a further 30 minutes. The organic phase was extracted and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure. The crude material was purified by column chromatography (silica gel, 0-10% MeOH in DCM). The title compound was obtained as a solid (17mg, 37% yield). LC/MS: calculated m/z value C₂₆H₂₄N₆OS([M+H]⁺): 469.5, measurement: 469.2.

examples I to 27

4-tert-butyl-N- { 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzyl } -benzamide

Step 1: { 2-fluoro-4- [7- (toluene-4-sulfonyl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzyl } -carbamic acid tert-butyl ester

4-chloro-7-tosyl-7H-pyrrolo [2,3-d ] combined in 5mL of water in a 20mL sealable microwave tube]Pyrimidine (1g, 3.25mmol, Eq: 1.00), [ 2-fluoro-4- (4, 4,5, 5-tetramethyl- [1, 3, 2-d-etramethyl- [1]Dioxolane-2-yl-benzyl]Tert-butyl carbamate (1.14g, 3.25mmol, Eq: 1.00) and potassium carbonate (1.8g, 13.0mmol, Eq: 4.00) with DME (10 mL). Adding Pd (PPh)₃)₄(375mg, 0.325mmol, Eq: 0.1). The reaction mixture was heated in a microwave at 150 ℃ for 30 minutes. The solution was washed with EtOAc and brine. The combined organic phases were dried over anhydrous sodium sulfate and the solvent was removed under reduced pressure. The crude material was purified by column chromatography (silica gel, 5-70% ethyl acetate in hexane). The title compound was obtained as a white solid (950mg, 59% yield). LC/MS: m/z meterCalculation of C₂₅H₂₅FN₄O₄S([M+H]⁺): 497.5, measurement: 497.2.

step 2: {4- [ 6-bromo-7- (toluene-4-sulfonyl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -2-fluoro-benzyl } -carbamic acid tert-butyl ester

In a 100mL round-bottom flask, { 2-fluoro-4- [7- (toluene-4-sulfonyl) -7H-pyrrolo [2,3-d]Pyrimidin-4-yl]-benzyl } -carbamic acid tert-butyl ester (2g, 4.03mmol, Eq: 1.00) was dissolved in THF (40mL) and cooled to-78 ℃. LDA 2M solution in heptane/THF/ethylbenzene (5.03mL, 10.1mmol, Eq: 2.5) was added under nitrogen at-78C to give a dark brown solution. The reaction mixture was stirred at-78 ℃ for 1hr 30 min. 1, 2-dibromo-1, 1,2, 2-tetrachloroethane (3.28g, 10.1mmol, Eq: 2.5) in 10mL of THF was added, and the reaction mixture was stirred at-78C for 2 hours. Brine was added. The reaction mixture was diluted with EtOAc and washed with brine. The combined organic phases were dried over anhydrous sodium sulfate and then evaporated. The crude material was purified by column chromatography (silica gel, 5-70% ethyl acetate in hexane). The title compound was obtained as a light brown solid (1.2g, 52% yield). LC/MS: calculated m/z value C₂₅H₂₄BrFN₄O₄S([M+H]⁺): 576.4, measurement: 577.1.

and step 3: { 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzyl } -carbamic acid tert-butyl ester

Combine {4- [ 6-bromo-7- (toluene-4-sulfonyl) -7H-pyrrolo [2,3-d ] in water (3mL) in a 20mL sealable microwave tube]Pyrimidin-4-yl]-2-fluoro-benzyl } -carbamic acid tert-butyl ester (400mg, 0.695mmol, Eq: 1.00),1-methyl-4- (4, 4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole (226mg, 1.09mmol, Eq: 1.6) and potassium carbonate (384mg, 2.78mmol, Eq: 4.00) with DME (6 mL). Adding Pd (PPh)₃)₄(80mg, 0.069mmol, Eq: 0.1), the reaction mixture was sealed and heated in a microwave at 160 ℃ for 60 minutes. The reaction mixture was diluted with EtOAc and then washed with brine. The combined organic phases were dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure. The crude material was triturated with DCM then filtered to give the title compound as a brown solid (75mg, 26% yield). LC/MS: calculated m/z value C₂₂H₂₃FN₆O₂([M+H]⁺): 423.4, measurement: 423.3.

and 4, step 4: 4-tert-butyl-N- { 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzyl } -benzamide

Combine { 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] in a 20mL scintillation vial]Pyrimidin-4-yl]-benzyl } -carbamic acid tert-butyl ester (40mg, 0.094mmol, Eq: 1.00) with 1mL DCM and 1mL TFA. The solution was stirred at room temperature for 1 hour. The solvent was removed under reduced pressure. The crude material was dissolved in DMF (2 mL). 4-tert-butylbenzoic acid (19mg, 0.104mmol, Eq: 1.1), DIPEA (0.066mL, 0.379mmol, Eq: 4.00) and HATU (40mg, 0.104mmol, Eq: 1.1) were added. The reaction mixture was stirred overnight. The resulting solution was diluted with 10mL of water and 5mL of EtOAc. The solution was stirred at room temperature for a further 30 minutes. The organic phase was extracted and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure. The resulting solid was triturated with DCM and the resulting solid was filtered. The title compound was obtained as a solid (17mg, 37% yield). LC/MS: calculated m/z value C₂₈H₂₇FN₆O([M+H]⁺): 483.5, measurement: 483.2.

examples I to 28

6-tert-butyl-N- { 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzyl } -nicotinamide

Step 1: 6-tert-butyl-nicotinic acid

To a suspension of nicotinic acid (2g, 16mmol, Eq: 1.00) in water was added concentrated sulfuric acid (1mL, 18.8mmol, Eq: 1.2), and the mixture was stirred under nitrogen atmosphere to form a clear solution. Pivalic acid (1.83g, 17.9mmol, Eq: 1.1) was added and stirring continued at ambient temperature for 10 min under an argon atmosphere. Silver nitrate (125mg, 0.736mmol) was added, then ammonium persulfate (295mg, 1.29mmol, Eq: 0.08) was added, the flask was wrapped with aluminum foil to protect from light, and the mixture was heated to 90 ℃ under a nitrogen atmosphere. After 2 hours the reaction mixture was cooled to ambient temperature and kept standing overnight. The reaction mixture was extracted with ethyl acetate, and it was difficult to obtain any of the expected products in large amounts. LC/MS did show the presence of product in the aqueous layer. The aqueous mixture was concentrated in vacuo to give a colorless solid. The solid was triturated with THF, filtered, and the filtrate concentrated in vacuo. The residue was triturated with methanol again, filtered and the filtrate was concentrated in vacuo. The concentrated filtrate was purified by reverse phase chromatography, eluting with a gradient from 10% acetonitrile in water to 100% acetonitrile using an 85g C-18 column. The fractions containing the desired product were combined and concentrated to give a colorless aqueous suspension (-5 mL volume). Water (20mL) was added to form a clear solution and the mixture was lyophilized to give the title compound as a colorless amorphous lyophilized solid (139mg, 5% yield). LC/MS: calculated m/z value C₁₀H₁₄NO₂[(M+H)⁺]: 180.2, 483.5, measurement: 180.1.

step 2: 6-tert-butyl-N- { 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzyl } -nicotinamide

Combine { 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] in a 20mL scintillation vial]Pyrimidin-4-yl]-benzyl } -carbamic acid tert-butyl ester (56mg, 0.133mmol, Eq: 1.00) with 2mL DCM and 2mL TFA. The solution was stirred at room temperature for 30 minutes. The solvent was removed under reduced pressure. The crude material was dissolved in DMF (2 mL). 6-tert-butyl-nicotinic acid (40mg, 0.233mmol, Eq: 1.7), DIPEA (0.093mL, 0.53mmol, Eq: 4.00) and HATU (55mg, 0.146mmol, Eq: 1.1) were added. The reaction mixture was stirred overnight. The resulting solution was diluted with 10mL of water and 5mL of EtOAc. The solution was stirred at room temperature for a further 30 minutes. The organic phase was extracted and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure. The resulting solid was triturated with DCM and filtered. The title compound was obtained as a yellow solid (40mg, 62% yield). LC/MS: calculated m/z value C₂₇H₂₆FN₇O([M+H]⁺): 484.5, measurement: 483.3.

examples I to 29

5-methyl-thiophene-2-carboxylic acid 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzylamide

According to a method similar to example 28, step 2, using 5-methylthiophene-2-carboxylic acid (22mg, 0.156mmol, Eq: 1.1), the title compound was obtained as a solid (42mg, 66% yield). LC/MS: calculated m/z value C₂₃H₁₉FN₆OS([M+H]⁺): 447.5, measured value：447.2。

Examples I to 30

4-tert-butyl-N- (2-fluoro-4- {6- [1- (2-hydroxy-ethyl) -1H-pyrazol-4-yl ] -7H-pyrrolo [2,3-d ] pyrimidin-4-yl } -benzyl) -benzamide

Step 1: n- [4- (6-bromo-7H-pyrrolo [2,3-d ] pyrimidin-4-yl) -2-fluoro-benzyl ] -4-tert-butyl-benzamide

In a 20mL scintillation vial, {4- [ 6-bromo-7- (toluene-4-sulfonyl) -7H-pyrrolo [2,3-d ] was combined]Pyrimidin-4-yl]-2-fluoro-benzyl } -carbamic acid tert-butyl ester (200mg, 0.348mmol, Eq: 1.00) with 1mL DCM and 1mL TFA. The solution was stirred at room temperature for 30 minutes. The solvent was removed under reduced pressure. The crude material was dissolved in DMF (2 mL). 4-tert-butyl-benzoic acid (68mg, 0.382mmol, Eq: 1.1), DIPEA (0.243mL, 1.39mmol, Eq: 4.00) and HATU (145mg, 0.382mmol, Eq: 1.1) were added. The reaction mixture was stirred overnight. The resulting solution was diluted with 10mL of water and 5mL of EtOAc. The solution was stirred at room temperature for a further 30 minutes. The organic phase was extracted and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure. The crude material was stored at room temperature overnight. The resulting solid was triturated with DCM and filtered. The title compound was obtained as a solid (96mg, 57% yield). LC/MS: calculated m/z value C₂₄H₂₂BrFN₄O([M+H]⁺): 482.3, measurement: 483.0. step 2: 4-tert-butyl-N- (2-fluoro-4- {6- [1- (2-hydroxy-ethyl) -1H-pyrazol-4-yl]-7H-pyrrolo [2,3-d]Pyrimidin-4-yl } -benzyl) -benzamides

Combining N- [4- (6-bromo-7H-pyrrolo [2, 3-d) in water (1mL) in a 20mL sealable microwave tube]Pyrimidin-4-yl) -2-fluoro-benzyl]-4-tert-butyl-benzamide (90mg, 0.187mmol, Eq: 1.00), 2- (4- (4, 4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazol-1-yl) ethanol (49.0mg, 0.206mmol, Eq: 1.1) and potassium carbonate (103mg, 0.748mmol, Eq: 4.00) with DME (4 mL). Adding Pd (PPh)₃)₄(22mg, 0.019mmol, Eq: 0.1), the reaction mixture was sealed and heated in a microwave at 150 ℃ for 30 minutes. The reaction mixture was diluted with EtOAc and then washed with brine. The combined organic phases were dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure. The crude material was triturated with DCM then filtered to give the title compound as a brown solid (22mg, 23% yield). LC/MS: calculated m/z value C₂₉H₂₉FN₆O₂([M+H]⁺): 513.5, measurement: 513.2.

examples I to 31

4-tert-butyl-N- { 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzyl } -N-methyl-benzamide

Step 1: (4-bromo-2-fluoro-benzyl) -methyl-carbamic acid tert-butyl ester

A60% oil suspension of tert-butyl 4-bromo-2-fluorobenzylcarbamate (2g, 6.58mmol, Eq: 1.00), methyl iodide (0.7mL, 11.2mmol, Eq: 1.7), and NaH (395mg, 16.5mmol, Eq: 2.5) was combined with DMF (40mL) at 0 deg.C in a 100mL round bottom flask. The reaction mixture was stirred and allowed to warm to room temperature for 3 hours. The reaction was stopped with MeOH. The reaction mixture was diluted with EtOAc and washed with brine. The combined organic phases were dried over anhydrous sodium sulfate and the solvent was removed under reduced pressure. The crude material was purified by column chromatography (silica gel, 5-40% ethyl acetate in hexane). The title compound was obtained as an oil (1.8g, 86% yield).

Step 2: [ 2-fluoro-4- (4, 4,5, 5-tetramethyl- [1, 3, 2] dioxaborolan-2-yl) -benzyl ] -methyl-carbamic acid tert-butyl ester

In a 250mL round bottom flask, (4-bromo-2-fluoro-benzyl) -methyl-carbamic acid tert-butyl ester (1.8g, 5.66mmol, Eq: 1.00), 4, 4, 4 ', 4 ', 5, 5, 5 ', 5 ' -octamethyl-2, 2 ' -bis (1, 3, 2-dioxaborolan) (2.15g, 8.49mmol, Eq: 1.5) and potassium acetate (1.67g, 17.0mmol, Eq: 3.00) were combined with NMP (40 mL). The solution was degassed for 10 minutes under a nitrogen atmosphere. 1, 1' -bis (diphenylphosphino) ferrocene ] -dichloropalladium (II) (414mg, 0.566mmol, Eq: 0.1) was added and the reaction mixture was heated at 100 ℃ for 24 hours. The reaction mixture was cooled and then diluted with water and EtOAc. The organic phases were combined and then dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure and the resulting crude material was purified by column chromatography (silica gel, 5-40% ethyl acetate in hexane). The title compound was obtained as an oil (1.01g, 49% yield).

And step 3: 7-benzenesulfonyl-4-chloro-6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidine

In a 20mL scintillation vial, combine 4-chloro-6-iodo-7- (phenylsulfonyl) -7H-pyrrolo [2,3-d ] in 5mL of water]Pyrimidines(1g, 2.38mmol, Eq: 1.00), potassium carbonate (1.32g, 9.53mmol, Eq: 4.00), and 1-methyl-4- (4, 4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole (744mg, 3.57mmol, Eq: 1.5) with DME (10 mL). Adding Pd (PPh)₃)₄(275mg, 0.238mmol, Eq: 0.1), the reaction mixture was heated at 90 ℃ for 6 hours. The reaction mixture was allowed to stand at room temperature overnight. A precipitate formed and was filtered under vacuum to give the title compound as a pale brown solid (194mg, 22% yield). LC/MS: calculated m/z value C₁₆H₁₂ClN₅O₂S([M+H]⁺): 374.8, measurement: 374.1.

and 4, step 4: { 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzyl } -methyl-carbamic acid tert-butyl ester

In a 20mL scintillation vial, 7-benzenesulfonyl-4-chloro-6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] was combined in 3mL of water]Pyrimidine (150mg, 0.401mmol, Eq: 1.00), [ 2-fluoro-4- (4, 4,5, 5-tetramethyl- [1, 3, 2-)]Dioxolane-2-yl-benzyl]Tert-butyl methyl-carbamate (161mg, 0.441mmol, Eq: 1.1) and potassium carbonate (222mg, 1.61mmol, Eq: 4.00) with DME (6 mL). Adding Pd (PPh)₃)₄(46.4mg, 0.04mmol, Eq: 0.1), the reaction mixture was heated at 160 ℃ for 60 minutes. The reaction mixture was diluted with EtOAc and washed with brine. The combined organic phases were dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure and the crude material was purified by column chromatography (silica gel, 5-70% ethyl acetate in hexane). The title compound was obtained as a solid (74mg, 42% yield). LC/MS: calculated m/z value C₂₃H₂₅FN₆O₂([M+H]⁺): 437.4, measurement: 437.3.

and 5: 4-tert-butyl-N- { 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzyl } -N-methyl-benzamide

Combine { 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] in a 20mL scintillation vial]Pyrimidin-4-yl]-benzyl } -methyl-carbamic acid tert-butyl ester (40mg, 0.092mmol, Eq: 1.00) with DCM (3mL) and TFA (3 mL). The solution was stirred at room temperature for 30 minutes. The solvent was removed under reduced pressure. The crude material was dissolved in DMF (4 mL). 4-tert-butyl-benzoic acid (18mg, 0.101mmol, Eq: 1.1), DIPEA (0.1mL, 0.573mmol, Eq: 6.25) and HATU (38mg, 0.101mmol, Eq: 1.1) were added. The reaction mixture was stirred overnight. The resulting solution was diluted with 10mL of water and 5mL of EtOAc. The solution was stirred at room temperature for a further 30 minutes. The organic phase was extracted and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure. The crude material was purified by column chromatography (silica gel, 0-10% MeOH in DCM). The title compound was obtained as a solid (38mg, 84% yield). LC/MS: calculated m/z value C₂₉H₂₉FN₆O([M+H]⁺): 497.5, measurement: 497.4.

examples I to 32

5-methyl-thiophene-2-carboxylic acid { 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzyl } -methyl-amide

Combine { 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] in a 20mL scintillation vial]Pyrimidin-4-yl]-benzyl } -methyl-carbamic acid tert-butyl ester (30mg, 0.069mmol, Eq: 1.00) with DCM (3mL) and TFA (3 mL). The solution was stirred at room temperature for 30 minutes. The solvent was removed under reduced pressure. The crude material was dissolved in DMF (4 mL). 5-methylthiophene-2-carboxylic acid (11mg, 0.076mmol, Eq: 1.1), DIPEA (0.05mL, 0.275mmol, Eq: 4.0) were added0) And HATU (29mg, 0.076mmol, Eq: 1.1). The reaction mixture was stirred overnight. The resulting solution was diluted with 10mL of water and 5mL of EtOAc. The solution was stirred at room temperature for a further 30 minutes. The organic phase was extracted and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure. The crude material was purified by column chromatography (silica gel, 0-10% MeOH in DCM). The title compound was obtained as a solid (14mg, 44% yield). LC/MS: calculated m/z value C₂₄H₂₁FN₆OS([M+H]⁺): 461.5, measurement: 461.2.

examples I to 33

2-tert-butyl-5- { 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzyl } -4, 5-dihydro-thieno [2,3-c ] pyrrol-6-one

Step 1: 3-methyl-thiophene-2-carboxylic acid methyl ester

In a 1L round bottom flask, combine 3-methylthiophene-2-carboxylic acid (15g, 106mmol) with methanol (211mL) to give a yellow-white suspension. The mixture was cooled to 0 ℃ in an ice-water bath. Concentrated sulfuric acid (6ml, 113mmol) was added dropwise to the cold suspension. The reaction mixture was gradually warmed to room temperature. The reaction mixture was stirred at room temperature for 3 days or more. After this time, TLC showed complete conversion of starting material to less polar product. The reaction mixture was concentrated to remove methanol. The remaining light brown oil was partitioned between ethyl acetate and saturated aqueous sodium bicarbonate. With Na₂SO₄The organic phase was dried, filtered and concentrated to give a brown oil based on¹H NMR integral, which comprises a mixture of the desired methyl ester (84%) and the starting material (16%). Will be coarseThe product was redissolved in ethyl acetate and the solution was washed with 1M aqueous NaOH. With MgSO₄The organic phase was dried, filtered and concentrated to give methyl 3-methyl-thiophene-2-carboxylate (13.6g, 82%) as a light brown oil.¹H NMR (300MHz, chloroform-d) ppm7.39(d, J ═ 5.09Hz, 1H), 6.92(d, J ═ 5.20Hz, 1H), 3.87(s, 3H), 2.57(s, 3H).

Step 2: 5-tert-butyl-3-methyl-thiophene-2-carboxylic acid methyl ester

In a 500mL round-bottom flask, aluminum trichloride (17.3g, 130mmol) was combined with DCM (20mL) to give an off-white suspension. The mixture was back-filled with argon and then cooled to-78 ℃ in a dry ice/acetone bath. A solution of methyl 3-methylthiophene-2-carboxylate (13.5g, 86.4mmol) in 10mL of DCM was added dropwise over 5 minutes. The reaction mixture was stirred at-78 ℃ for 5 minutes. A solution of 2-chloro-2-methylpropane (9.87mL, 90.7mmol) in 10mL DCM was added dropwise over 30min to the cold reaction mixture. The reaction mixture was stirred under a reflux condenser over the course of 1 week using a dry ice/acetone bath, gradually melted, and the reaction flask warmed to room temperature. The reaction mixture was poured into ice water. After melting the ice, the organic phase is separated and then treated with Na₂SO₄And (5) drying. The organic phase was filtered and then concentrated to give a brown oil. The oil was applied directly to a 330 g silica gel column. Flash chromatography (0-5% EtOAc-hexanes) was used to isolate 5-tert-butyl-3-methyl-thiophene-2-carboxylic acid methyl ester (7.05g, 38%) as a yellow oil.¹H NMR (300MHz, chloroform-d) ppm 6.68(s, 1H), 3.84(s, 3H), 2.50(s, 3H), 1.38(s, 9H).

And step 3: 3-Bromomethyl-5-tert-butyl-thiophene-2-carboxylic acid methyl ester

In a 1L pear-shaped flask, methyl 5-tert-butyl-3-methylthiophene-2-carboxylate (6.06g, 28.5mmol), N-bromosuccinimide (6.1g, 34.3mmol), and azobisisobutyronitrile (234mg, 1.43mmol) were combined with carbon tetrachloride (80mL) to give an orange suspension. The mixture was heated at 90 ℃ overnight. In the morning, the reaction mixture was cooled to room temperature and then filtered to remove precipitated solids. The filtrate was concentrated to give a brown oil. The product was directly applied to a 120g silica gel column. Flash chromatography (5% EtOAc-hexanes) gave only partially purified product. The fractions containing pure product were combined to give methyl 3-bromomethyl-5-tert-butyl-thiophene-2-carboxylate (2.65g, 32%) as a yellow oil. The impure fraction from above was applied to a second column, followed by 120g of silica gel and 5% EtOAc-hexane). Another batch of purified methyl 3-bromomethyl-5-tert-butyl-thiophene-2-carboxylate (2.54g, 30%) was obtained.¹H NMR (300MHz, chloroform-d) ppm 6.93(s, 1H), 4.87(s, 2H), 3.88(s, 3H), 1.39(s, 8H).

And 4, step 4: 3- [ (4-bromo-2-fluoro-benzylamino) -methyl ] -5-tert-butyl-thiophene 2-carboxylic acid methyl ester

In a 250mL round bottom flask, combine 4-bromo-2-fluoro-benzylamine (5.34g, 26.2mmol), 3-bromomethyl-5-tert-butyl-thiophene-2-carboxylic acid methyl ester (2.54g, 8.72mmol), and cesium carbonate (3.73g, 11.4mmol) with acetonitrile (50mL) to give a white suspension. The reaction mixture was stirred at room temperature over the weekend. The reaction mixture was filtered, and the filtrate was concentrated with a rotary evaporator. The crude product was directly applied to a 120g silica gel column. Flash chromatography (5-25% EtOAc-hexanes) afforded 3- [ (4-bromo-2-fluoro-benzylamino) -methyl]Methyl-5-tert-butyl-thiophene-2-carboxylate (1.88g, 52%) as a light yellow oil. LC/MS: calculated m/z value C₁₈H₂₂BrFN([M+H]⁺): 414 and 416, measured values: 416.0.

and 5: 3- [ (4-bromo-2-fluoro-benzylamino) -methyl ] -5-tert-butyl-thiophene-2-carboxylic acid

In a 1L pear flask, combine 3- [ (4-bromo-2-fluoro-benzylamino) -methyl]Methyl-5-tert-butyl-thiophene-2-carboxylate (1.85g, 4.47mmol) and lithium hydroxide monohydrate (1.87g, 44.7mmol) with THF (12mL) and water (12mL) gave a colorless suspension. The mixture was stirred at room temperature overnight. In the morning, LCMS showed most starting material and a small amount of product. Methanol (5mL) was added and the reaction mixture was heated at 50 ℃ for 20 hours. The reaction mixture was cooled to room temperature and concentrated to dryness on a rotary evaporator. The resulting yellow-white solid was partially dissolved in water and then 4N aqueous HCl was added until the mixture became a white suspension. The suspension was extracted with ethyl acetate. The organic phase was dried (Na)₂SO₄) Filtered and then concentrated to give 3- [ (4-bromo-2-fluoro-benzylamino) -methyl]-5-tert-butyl-thiophene-2-carboxylic acid (1.77g, 99%) as an off-white foam. LC/MS: calculated m/z value C₁₇H₂₀BrFNO([M+H]⁺): 400 and 402, measurement values: 402.0.

step 6: 5- (4-bromo-2-fluoro-benzyl) -2-tert-butyl-4, 5-dihydro-thieno [2,3-c ] pyrrol-6-one

In a 1L round bottom flask, combine 3- ((4-bromo-2-fluorobenzylamino) methyl) -5-tert-butylthiophene-2-carboxylic acid (1.77g, 4.42mmol) with dichloromethane (80mL) to give a light yellow solution. The reaction flask was back-filled with argon, and thionyl chloride (1.96g, 1.2mL, 16.4mmol) was then added dropwise over 5 minutes. The reaction mixture was stirred at room temperature under an argon atmosphere for 18 hours. Thereafter, LCMS showed a mixture of starting material and product. 1.5mL of thionyl chloride was added to the reaction mixture, and the reaction mixture was stirred at room temperature for another 24 hours. Thereafter, LCMS showsThe reaction was complete. The reaction mixture was concentrated to give a pale yellow oil. The crude product was dissolved in dichloromethane and the resulting solution was concentrated on silica gel. The crude product supported on silica gel was loaded onto a 120g silica gel column. Flash chromatography (5-25% EtOAc-hexanes) afforded 5- (4-bromo-2-fluoro-benzyl) -2-tert-butyl-4, 5-dihydro-thieno [2, 3-c)]Pyrrol-6-one (1.22g, 72%) as a light yellow oil. LC/MS: calculated m/z value C₁₇H₁₈BrFNOS([M+H]⁺): 382 and 384, measured values: 384.0.

and 7: 2-tert-butyl-5- [ 2-fluoro-4- (4, 4,5, 5-tetramethyl- [1, 3, 2] dioxaborolan-2-yl) -benzyl ] -4, 5-dihydro-thieno [2,3-c ] pyrrol-6-one

In a 250mL round bottom flask, bis (pinacolato) diboron (1.15g, 4.53mmol), 5- (4-bromo-2-fluoro-benzyl) -2-tert-butyl-4, 5-dihydro-thieno [2,3-c ] were combined]Pyrrol-6-one (1.07g, 2.8mmol) and potassium acetate (825mg, 8.41mmol) with dioxane (9mL) gave a dark brown suspension. To the mixture was added 1, 1' -bis (diphenylphosphino) ferrocene-palladium (II) dichloride dichloromethane complex (185mg, 227. mu. mol). The reaction mixture was heated at 110 ℃ for 8 hours. After this time, the reaction mixture was cooled to room temperature and the dioxane was evaporated. The crude product was dissolved in dichloromethane and the solution was poured into water (30 mL). The organic phase is separated and then MgSO₄Dried, filtered and concentrated on silica gel. The crude product supported on silica gel was applied to a 120g silica gel column. Flash chromatography (5-25% ethyl acetate in hexane) afforded 2-tert-butyl-5- [ 2-fluoro-4- (4, 4,5, 5-tetramethyl- [1, 3, 2)]Dioxolane-2-yl-benzyl]-4, 5-dihydro-thieno [2,3-c]Pyrrol-6-one (0.72g, 60%) as a white powder. LC/MS: calculated m/z value C₂₃H₃₀BFNO₃S([M+H]⁺): 430, measured value: 430.2.

and 8: 2-tert-butyl-5- { 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzyl } -4, 5-dihydro-thieno [2,3-c ] pyrrol-6-one

Combine 7-benzenesulfonyl-4-chloro-6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] in 2mL of water in a 10mL microwave tube]Pyrimidine (150mg, 0.401mmol, Eq: 1.00), 2-tert-butyl-5- [ 2-fluoro-4- (4, 4,5, 5-tetramethyl- [1, 3, 2)]Dioxolane-2-yl-benzyl]-4, 5-dihydro-thieno [2,3-c]Pyrrol-6-one (190mg, 0.441mmol, Eq: 1.1) and potassium carbonate (222mg, 1.61mmol, Eq: 4.00) with DME (4 mL). Adding Pd (Ph)₃P)₄(46mg, 0.04mmol, Eq: 0.1). The reaction mixture was heated in a microwave at 160 ℃ for 60 minutes. The solution was diluted with EtOAc and then washed with brine. The combined organic phases were dried over anhydrous sodium sulfate and the solvent was removed under reduced pressure. The crude material was purified by column chromatography (silica gel, 0-10% MeOH in DCM). The title compound was obtained as a pale yellow solid (17mg, 9% yield). LC/MS: calculated m/z value C₂₇H₂₅FN₆OS([M+H]⁺): 501.6, measurement: 501.3.

examples I to 34

5-tert-butyl-isoxazole-3-carboxylic acid 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzylamide

Combine { 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] in a 20mL scintillation vial]Pyrimidin-4-yl]-benzyl } -methyl-carbamic acid tert-butyl ester (75mg, 0.178mmol, Eq: 1.00) with DCM (3mL) and TFA (3 mL). The solution was stirred at room temperature for 30 minutes. The solvent was removed under reduced pressure.The crude material was dissolved in DMF (4 mL). 5-tert-butylisoxazole-3-carboxylic acid (18mg, 0.195mmol, Eq: 1.1), DIPEA (0.124mL, 0.710mmol, Eq: 4.00) and HATU (74mg, 0.195mmol, Eq: 1.1) were added. The reaction mixture was stirred overnight. The resulting solution was diluted with 10mL of water and 5mL of EtOAc. The solution was stirred at room temperature for a further 30 minutes. The organic phases were combined and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure. The crude material was purified by column chromatography (silica gel, 0-10% MeOH in DCM). The title compound was obtained as a yellow solid (35mg, 42% yield). LC/MS: calculated m/z value C₂₅H₂₄FN₇O₂([M+H]⁺): 474.5, measurement: 474.3.

examples I to 35

N- { 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzyl } -4- (3-methyl-oxetan-3-yl) -benzamide

Combine { 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] in a 10mL sealable tube]Pyrimidin-4-yl]-benzyl } -methyl-carbamic acid tert-butyl ester (65mg, 0.155mmol, Eq: 1.00) with DCM (3mL) and TFA (3 mL). The solution was stirred at room temperature for 60 minutes. The solvent was removed under reduced pressure and dried in vacuo. The crude material was dissolved in DMF (5 mL). 4- (3-Methylooxetan-3-yl) benzoic acid (59.6mg, 0.31mmol, Eq: 2.00), DIPEA (0.14mL, 0.78mmol, Eq: 5.00) and HATU (118mg, 0.31mmol, Eq: 2.00) were added. The reaction mixture was stirred at room temperature overnight. The resulting solution was diluted with EtOAc, washed with water and brine. The combined organic phases were dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure. The crude material was purified by column chromatography (silica gel, 0-10% MeOH in DCM). The title compound was obtained as a solid (44mg, 57% yield). LC/MS: calculated m/z value C₂₈H₂₅FN₆O₂([M+H]⁺): 497.5, measurement: 497.2.

examples I to 36

4- (cyano-dimethyl-methyl) -N- { 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzyl } -benzamide

According to a method similar to example 35, using 4- (2-cyanoprop-2-yl) benzoic acid (58.7mg, 0.310mmol, Eq: 2.00), the title compound was obtained as a solid (43mg, 53% yield). LC/MS: calculated m/z value C₂₈H₂₄FN₇O([M+H]⁺): 494.5, measurement: 494.2.

examples I to 37

4,5,6, 7-tetrahydro-benzo [ b ] thiophene-2-carboxylic acid 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzylamide

In a similar manner to example 35, 4,5,6, 7-tetrahydrobenzo [ b ] was used]Thiophene-2-carboxylic acid (57mg, 0.31mmol, Eq: 2.00) to give the title compound as a solid (53mg, 63% yield). LC/MS: calculated m/z value C₂₆H₂₃FN₆OS([M+H]⁺): 487.5, measurement: 487.2.

examples I to 38

N- { 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzyl } -4- (1-hydroxy-1-methyl-ethyl) -benzamide

Step 1: 4- (1-hydroxy-1-methyl-ethyl) -benzoic acid

In a 500mL round bottom flask, combine 4-isopropylbenzoic acid (1.0g, 6.09mmol) with 5mL 10% aqueous KOH to give a cloudy suspension. An aqueous solution of KOH (96mL, 19.2mmol) and potassium permanganate (1.92g, 12.2mmol) in 100mL of water were added. The reaction mixture was heated at 70 ℃ for 1 hour. To the reaction mixture was added 5 drops of glycerol. The reaction mixture was cooled to 0 ℃. The solid residue was filtered through a pad of celite. The filtrate was washed 2 times with diethyl ether. The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, concentrated and dried overnight. The product, 4- (1-hydroxy-1-methyl-ethyl) -benzoic acid, was collected as a white solid (870mg, 79%) which was used in the next step without further purification.¹H NMR(400MHz，DMSO-d₆)ppm 12.80(br.s，1H)，7.83-7.89(m，2H)，7.55-7.60(m，2H)，5.15(s，1H)，1.43(s，6H)。

Step 2: n- { 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzyl } -4- (1-hydroxy-1-methyl-ethyl) -benzamide

According to a method similar to example 35, using 4- (2-hydroxyprop-2-yl) benzoic acid (41.4mg, 0.230mmol, Eq: 2.00), the title compound was obtained as a solid (46mg, 82% yield). LC/MS: calculated m/z value C₂₇H₂₅FN₆O₂([M+H]⁺): 485.5, measurement: 485.4.

examples I to 39

Step 1: 3-tert-butyl-isoxazole-5-carboxylic acid methyl ester

To a solution of pivalaldehyde (1.0g, 11.6mmol) in 1: 1 t-butanol/water (40mL) was added hydroxylamine hydrochloride (807mg, 11.6mmol) and sodium hydroxide (464mg, 11.6 mmol). The mixture was stirred at room temperature for 30 minutes, then chloramine-T (5.49g, 23.4mmol) was added in portions over 5 minutes, followed by copper (II) sulfate (327mg, 1.3mmol) and copper powder (73.8mg, 1.16mmol) and methyl propiolate (976mg, 11.6 mmol). The reaction mixture was heated at reflux where it was maintained for 2 h. After this time, the mixture was cooled to room temperature and poured into ice/water (50 g). Ammonium hydroxide (10mL) was added and the solution was extracted with DCM (3 × 200 mL). The organic layers were combined and dried (Na)₂SO₄) Filtering, and concentrating under reduced pressure. The crude material was purified by flash chromatography (silica gel, 40g, 0% to 10% EtOAc in hexanes) to give 3-tert-butyl-isoxazole-5-carboxylic acid methyl ester (427mg, 20%) as a colorless oil. LC/MS: calculated m/z value C₉H₁₃NO₃[(M+H)⁺]184, measurement value: 184.1.

step 2: 3-tert-butyl-isoxazole-5-carboxylic acid

To a solution of methyl 3-tert-butyl-isoxazole-5-carboxylate (425mg, 2.32mmol) in methanol (4mL) was added aqueous 1N NaOH (11.6mL, 11.6 mmol). The reaction mixture was stirred at room temperature for 2 hours, then concentrated and neutralized with 1N hydrochloric acid (10 mL). The mixture was extracted with ethyl acetate, dried over sodium sulfate, filtered, and concentrated to give 3-tert-butyl-isoxazole-5-carboxylic acid (318mg, 81%) as a white semi-solid. LC/MS: calculated m/z value C₈H₁₁NO₃[(M+H)⁺]170, measured value: 170.

and step 3: 3-tert-butyl-isoxazole-5-carboxylic acid 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzylamide

According to a method similar to example 35, using 3-tert-butylisoxazole-5-carboxylic acid (39mg, 0.230mmol, Eq: 2.00), the title compound was obtained as a solid (39mg, 71% yield). LC/MS: calculated m/z value C₂₅H₂₄FN₇O₂([M+H]⁺): 474.5. measurement value: 474.3.

examples I to 40

3-tert-butoxy-azetidine-1-carboxylic acid 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzylamide

Combine { 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] in a 20mL sealable microwave tube]Pyrimidin-4-yl]-benzyl } -methyl-carbamic acid tert-butyl ester (65mg, 0.155mmol, Eq: 1.00)With DCM (3mL) and TFA (3 mL). The solution was stirred at room temperature for 60 minutes. The solvent was removed under reduced pressure and dried in vacuo. The crude material was dissolved in DMF (4 mL). Bis (1H-imidazol-1-yl) methanone (50.3mg, 0.31mmol, Eq: 2.00) and DIPEA (0.14mL, 0.78mmol, Eq: 5.00) were added to the reaction mixture to give a light yellow solution. The reaction was stirred at room temperature for 2 hrs. 3-tert-Butoxyazetidine (40mg, 0.310mmol, Eq: 2.00) was added and stirred at room temperature overnight. The reaction mixture was diluted with EtOAc and washed with brine. The organic phases were combined and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure. The crude material was purified by column chromatography (silica gel, 0-10% MeOH in DCM). The title compound was obtained as a yellow solid (32mg, 43% yield). LC/MS: calculated m/z value C₂₅H₂₈FN₇O₂([M+H]⁺): 478.5, measurement: 478.6.

examples I to 41

1, 3-dihydro-isoindole-2-carboxylic acid 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzylamide

By a method similar to example 40, using isoindoline (37.0mg, 0.31mmol, Eq: 2.00), the title compound was obtained as a solid (43mg, 59% yield). LC/MS: calculated m/z value C₂₆H₂₂FN₇O([M+H]⁺): 468.5, measurement: 468.1.

example 42

4-tert-butyl-N- (4- {6- [1- (2-dimethylamino-ethyl) -1H-pyrazol-4-yl ] -7H-pyrrolo [2,3-d ] pyrimidin-4-yl } -2-fluoro-benzyl) -benzamide

Step 1: n, N-dimethyl-2- (4- (4, 4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazol-1-yl) -ethylamine

In a 250mL round bottom flask, 2-chloro-N, N-dimethylethylamine (998mg, 9.28mmol), 4- (4, 4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole (1.2g, 6.18mmol), and cesium carbonate (4.03g, 12.4mmol) were combined with acetonitrile (20mL) to give a white suspension. The reaction mixture was heated at 100 ℃ overnight. In the morning, the reaction mixture was cooled to room temperature, filtered, and the filtrate was concentrated to give the title compound (1.26g, 77% yield) as a colorless oil. The crude product was used for the subsequent reaction without further purification.

Step 2: (4- {6- [1- (2-dimethylamino-ethyl) -1H-pyrazol-4-yl ] -7H-pyrrolo [2,3-d ] pyrimidin-4-yl } -2-fluorobenzyl) -carbamic acid tert-butyl ester

4- (6-bromo-7-tosyl-7H-pyrrolo [2,3-d ] combined in a 20mL sealable microwave tube]Pyrimidin-4-yl) -2-fluorobenzylcarbamic acid tert-butyl ester (250mg, 0.434mmol, Eq: 1.00), N-dimethyl-2- (4- (4, 4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazol-1-yl) ethylamine (319mg, 1.2mmol, Eq: 2.77) and Pd (PPh)₃)₄(50mg, 0.043mmol, Eq: 0.1) with DME (4ml) gave a light brown suspension. Water (1mL) was added followed by potassium carbonate (240mg, 1.74mmol, Eq: 4.00). The reaction mixture was heated at 150 ℃ for 1hr with a microwave. The reaction mixture was diluted with EtOAc, washed with brine and water. The organic phases are combined withDried over anhydrous sodium sulfate. The solvent was removed under reduced pressure. The crude material was purified by column chromatography (silica gel, 0-10% MeOH in DCM). The title compound was obtained as a yellow solid (62mg, 29% yield). LC/MS: calculated m/z value C₂₅H₃₀FN₇O₂([M+H]⁺): 480.6, measurement: 480.3.

and step 3: (2- {4- [4- (4-aminomethyl-3-fluoro-phenyl) -7H-pyrrolo [2,3-d ] pyrimidin-6-yl ] -pyrazol-1-yl } -ethyl) -dimethyl-amine

In a 100mL round-bottom flask, combine 4- (6- (1- (2- (dimethylamino) ethyl) -1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d]Pyrimidin-4-yl) -2-fluorobenzylcarbamic acid tert-butyl ester (62mg, 0.129mmol, Eq: 1.00) with DCM (8mL) to give a pale yellow suspension. TFA (4mL, 51.9mmol, Eq: 402) was added and stirred at room temperature for 1 hr. The solvent was removed under reduced pressure. The crude material was dried under high vacuum for 2 hrs. The residue was used in the next step without further purification. LC/MS: calculated m/z value C₂₀H₂₂FN₇([M+H]⁺): 380.4, measurement: 380.2.

and 4, step 4: 4-tert-butyl-N- (4- {6- [1- (2-dimethylamino-ethyl) -1H-pyrazol-4-yl ] -7H-pyrrolo [2,3-d ] pyrimidin-4-yl } -2-fluoro-benzyl) -benzamide

(2- {4- [4- (4-aminomethyl-3-fluoro-phenyl) -7H-pyrrolo [2,3-d ] was combined in a 10mL sealable tube]Pyrimidin-6-yl]-pyrazol-1-yl } -ethyl) -dimethyl-amine (48mg, 0.127mmol, Eq: 1.00), 4-tert-butylbenzoic acid (45mg, 0.253mmol, Eq: 2.00) and HATU (96mg, 0.253mmol, Eq: 2.00) with DMF (4mL) to give a yellow solution. The reaction mixture was stirred for 5 minutes, then DIPEA (0.110ml, 0) was added.630mmol, Eq: 5.00), stirred at room temperature overnight. The reaction mixture was diluted with EtOAc and washed with brine. The organic phases were combined and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure. The crude material was purified by column chromatography (silica gel, 0-10% MeOH in DCM). The title compound was obtained as a solid (49mg, 71% yield). LC/MS: calculated m/z value C₃₁H₃₄FN₇O([M+H]⁺): 540.6, measurement: 540.3.

examples I to 43

3-tert-butoxy-azetidine-1-carboxylic acid 4- {6- [1- (2-dimethylamino-ethyl) -1H-pyrazol-4-yl ] -7H-pyrrolo [2,3-d ] pyrimidin-4-yl } -2-fluoro-benzylamide

(2- {4- [4- (4-aminomethyl-3-fluoro-phenyl) -7H-pyrrolo [2,3-d ] was combined in a 20mL sealable microwave tube]Pyrimidin-6-yl]-pyrazol-1-yl } -ethyl) -dimethyl-amine (22mg, 0.056mmol, Eq: 1.00), bis (1H-imidazol-1-yl) methanone (19mg, 0.116mmol, Eq: 2.00) and DIPEA (51 μ l, 0.290mmol, Eq: 5.00) with DMF (2mL) to give a pale yellow solution. The reaction was stirred at room temperature for 2 hrs. 3-tert-Butoxyazetidine (15.0mg, 0.116mmol, Eq: 2.00) was added and stirred at room temperature overnight. The reaction mixture was diluted with EtOAc, washed with brine and water. The organic phases were combined and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure. The crude material was purified by column chromatography (silica gel, 0-10% MeOH in DCM). The title compound was obtained as a yellow solid (18mg, 58% yield). LC/MS: calculated m/z value C₂₈H₃₅FN₈O₂([M+H]⁺): 535.6. measurement value: 535.4.

examples I to 44

1, 3-dihydro-isoindole-2-carboxylic acid 4- {6- [1- (2-dimethylamino-ethyl) -1H-pyrazol-4-yl ] -7H-pyrrolo [2,3-d ] pyrimidin-4-yl } -2-fluoro-benzylamide

By a method similar to example 43, using isoindoline (14mg, 0.116mmol, Eq: 2.00), the title compound was obtained as a yellow solid (20mg, 63% yield). LC/MS: calculated m/z value C₂₉H₂₉FN₈O([M+H]⁺): 525.6, measurement: 525.3.

examples I to 45

[4- (4- {4- [ (4-tert-butyl-benzoylamino) -methyl ] -3-fluoro-phenyl } -7H-pyrrolo [2,3-d ] pyrimidin-6-yl) -pyrazol-1-yl ] -acetic acid ethyl ester

Step 1: (4- {4- [4- (tert-Butoxycarbonylamino-methyl) -3-fluoro-phenyl ] -7H-pyrrolo [2,3-d ] pyrimidin-6-yl } -pyrazol-1-yl) -acetic acid ethyl ester

4- (6-bromo-7-tosyl-7H-pyrrolo [2,3-d ] combined in a 20mL sealable microwave tube]Pyrimidin-4-yl) -2-fluorobenzylcarbamic acid tert-butyl ester (250mg, 0.434mmol, Eq: 1.00), ethyl 2- (4- (4, 4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazol-1-yl) acetate (365mg, 1.3mmol, Eq: 3) and Pd (PPh)₃)₄(50mg, 0.043mmol, Eq: 0.1) with DMF (10mL) gave a light brown suspension. Potassium carbonate (240mg, 1.74mmol, Eq: 4.00) was added. Reacting the mixtureThe mixture was heated at 155 deg.C for 1hr with a microwave. The reaction mixture was diluted with EtOAc, washed with brine and water. The organic phases were combined and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure. The crude material was purified by column chromatography (silica gel, 5-60% EtOAc in hexanes). The title compound was obtained as a solid (72mg, 33% yield). LC/MS: calculated m/z value C₂₅H₂₇FN₆O₄([M+H]⁺): 495.5, measurement: 495.3.

step 2: {4- [4- (4-aminomethyl-3-fluoro-phenyl) -7H-pyrrolo [2,3-d ] pyrimidin-6-yl ] -pyrazol-1-yl } -acetic acid ethyl ester

In a 50mL round bottom flask, (4- {4- [4- (tert-butoxycarbonylamino-methyl) -3-fluoro-phenyl) -was combined]-7H-pyrrolo [2,3-d]Pyrimidin-6-yl } -pyrazol-1-yl) -acetic acid ethyl ester (72mg, 0.146mmol, Eq: 1.00) with DCM (8mL) to give a yellow solution. TFA (4mL, 51.9mmol, Eq: 357) was added and stirred at room temperature for 1 hr. The solvent was removed under reduced pressure and dried under vacuum for 3 hrs. The residue was used in the next step without further purification. LC/MS: calculated m/z value C₂₀H₁₉FN₆O₂([M+H]⁺): 395.4, measurement: 395.2.

and step 3: [4- (4- {4- [ (4-tert-butyl-benzoylamino) -methyl ] -3-fluoro-phenyl } -7H-pyrrolo [2,3-d ] pyrimidin-6-yl) -pyrazol-1-yl ] -acetic acid ethyl ester

In a 50mL round-bottom flask, {4- [4- (4-aminomethyl-3-fluoro-phenyl) -7H-pyrrolo [2,3-d ] is combined]Pyrimidin-6-yl]-pyrazol-1-yl } -acetic acid ethyl ester (52mg, 0.132mmol, Eq: 1.00), 4-tert-butylbenzoic acid (47.0mg, 0.264mmol, Eq: 2.00) and HATU (100mg, 0.264mmol, Eq: 2.00) with DMF (4mL) gave yellowA colored solution. The reaction mixture was stirred for 5 minutes, then DIPEA (115. mu.l, 659. mu. mol, Eq: 5.00) was added and stirred at room temperature overnight. The reaction mixture was diluted with EtOAc, washed with brine and water. The organic phases were combined and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure. The crude material was purified by column chromatography (silica gel, 0-10% MeOH in DCM). The title compound was obtained as a solid (52mg, 69% yield). LC/MS: calculated m/z value C₃₁H₃₁FN₆O₃([M+H]⁺): 555.6, measurement: 555.4.

example 46

[4- (4- {4- [ (4-tert-butyl-benzoylamino) -methyl ] -3-fluoro-phenyl } -7H-pyrrolo [2,3-d ] pyrimidine-6-

In a 50mL round-bottom flask, [4- (4- {4- [ (4-tert-butyl-benzoylamino) -methyl ] -was combined]-3-fluoro-phenyl } -7H-pyrrolo [2,3-d]Pyrimidin-6-yl) -pyrazol-1-yl]Ethyl acetate (47mg, 0.085mmol, Eq: 1.00) with THF (5mL) gave a yellow suspension. NaOH 1M solution (0.135ml, 0.135mmol, Eq: 1.59) was added and stirred at room temperature overnight. The reaction was acidified by addition of 1N HCl. The solvent was removed under reduced pressure. The residue was purified by HPLC to give the title compound as a yellow solid (34mg, 76% yield). LC/MS: calculated m/z value C₂₉H₂₇FN₆O₃([M+H]⁺): 527.6, measurement: 527.3.

example 47

N- (2-fluoro-4- (6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl) benzyl) -4,5,6, 7-tetrahydropyrazolo [1,5-a ] pyridine-2-carboxamide

Step 1: n- (4-bromo-2-fluorobenzyl) -4,5,6, 7-tetrahydropyrazolo [1,5-a ] pyridine-2-carboxamide

(4-bromo-2-fluorophenyl) methylamine (193mg, 945. mu. mol, Eq: 1.00), 4,5,6, 7-tetrahydropyrazolo [1, 5-a) in DMF (3.15ml)]Pyridine-2-carboxylic acid (157mg, 945. mu. mol, Eq: 1.00), HBTU (358mg, 945. mu. mol, Eq: 1.00) and DIPEA (366mg, 495. mu.l, 2.83mmol, Eq: 3) were stirred at r.t. for 16 h. The reaction mixture was diluted with ethyl acetate, washed with water and brine. The combined organic layers were dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure. The resulting crude material was purified by column chromatography (silica gel, 10-65% ethyl acetate in hexane) to give N- (4-bromo-2-fluorobenzyl) -4,5,6, 7-tetrahydropyrazolo [1,5-a ]]Pyridine-2-carboxamide (178mg, 54% yield) as a colorless oil. LC/MS: calculated m/z value C₁₅H₁₅BrFN₃O([M+H]⁺): 353.2, measurement: 354.1. step 2: n- (2-fluoro-4- (4, 4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) benzyl) -4,5,6, 7-tetrahydropyrazolo [1,5-a ]]Pyridine-2-carboxamides

N- (4-bromo-2-fluorobenzyl) -4,5,6, 7-tetrahydropyrazolo [1,5-a ] in NMP (3mL)]Pyridine-2-carboxamide (178mg, 505. mu. mol, Eq: 1.00), 4, 4, 4 ', 4 ', 5, 5, 5 ', 5 ' -octamethyl-2, 2 ' -bis (1, 3, 2-dioxaborolan) (193mg, 758. mu. mol, Eq: 1.5), PdCl₂(dppf)-CH₂Cl₂The adduct (37.0mg, 50.5. mu. mol, Eq: 0.1) and potassium acetate (149mg, 1.52mmol, Eq: 3) were heated to 100 ℃ for 16 h. By usingThe reaction mixture was diluted with ethyl acetate and washed with water and brine. The combined organic layers were dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure. The resulting crude material was purified by column chromatography (silica gel, 30-100% ethyl acetate in hexane) to give N- (2-fluoro-4- (4, 4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) benzyl) -4,5,6, 7-tetrahydropyrazolo [1,5-a ] benzyl ] -4,5,6, 7-tetrahydropyrazolo [1,5-a ] e]Pyridine-2-carboxamide (74mg, 37% yield) as a white solid. LC/MS: calculated m/z value C₂₁H₂₇BFN₃O₃([M+H]⁺): 400.2, measurement: 400.2.

and step 3: n- (2-fluoro-4- (6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl) benzyl) -4,5,6, 7-tetrahydropyrazolo [1,5-a ] pyridine-2-carboxamide

4-chloro-6- (1-methyl-1H-pyrazol-4-yl) -7- (phenylsulfonyl) -7H-pyrrolo [2,3-d ] was reacted in DME (1.48 ml)/water (371. mu.l)]Pyrimidine (69.3mg, 185. mu. mol, Eq: 1.00), N- (2-fluoro-4- (4, 4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) benzyl) -4,5,6, 7-tetrahydropyrazolo [1,5-a ] benzyl]Pyridine-2-carboxamide (74mg, 185. mu. mol, Eq: 1.00), tetrakis (triphenylphosphine) palladium (0) (21.4mg, 18.5. mu. mol, Eq: 0.1) and potassium carbonate (76.8mg, 556. mu. mol, Eq: 3) were heated to 150 ℃ for 45min in a microwave. Purification by column chromatography (silica gel, 0-100% ethyl acetate [ 10% MeOH/ethyl acetate)]Solution) and then purified by HPLC to give N- (2-fluoro-4- (6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2, 3-d)]Pyrimidin-4-yl) benzyl) -4,5,6, 7-tetrahydropyrazolo [1,5-a]Pyridine-2-carboxamide (5.7mg, 7% yield) as a yellow solid.¹H NMR(400MHz，DMSO-d)ppm 12.63(s，1H)，8.79(s，1H)，8.73(t，J＝5.8Hz，1H)，8.31(s，1H)，8.11(s，1H)，8.04(d，J＝7.8Hz，1H)，7.94(d，J＝11.5Hz，1H)，7.52(t，J＝8.2Hz，1H)，7.15(s，1H)，6.43(s，1H)，4.56(d，J＝6.2Hz，2H)，4.15(t，J＝5.3Hz，2H)，3.92(s，3H)，2.80(t, J ═ 5.0Hz, 2H), 2.01(br.s, 2H), 1.82(br.s, 2H); LC/MS: calculated m/z value C₂₅H₂₃FN₈O([M+H]⁺): 471.5, measurement: 471.2.

example 48

Combine { 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] in a 20mL scintillation vial]Pyrimidin-4-yl]-benzyl } -carbamic acid tert-butyl ester (75mg, 0.178mmol, Eq: 1.00) with 2mL DCM and 2mL TFA. The solution was stirred at room temperature for 30 minutes. The solvent was removed under reduced pressure. The crude material was dissolved in DMF (2 mL). 5-tert-butylisoxazole-3-carboxylic acid (33.0mg, 0.195mmol, Eq: 1.1), DIPEA (0.124mL, 0.71mmol, Eq: 4.00) and HATU (74mg, 0.195mmol, Eq: 1.1) were added. The reaction mixture was stirred overnight. The resulting solution was diluted with 10mL of water and 5mL of LEtOAc. The solution was stirred at room temperature for a further 30 minutes. The organic phase was extracted and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure. The resulting solid was purified by column chromatography (silica gel, 0-10% MeOH in DCM). The title compound was obtained as a yellow solid (35mg, 42% yield). LC/MS: calculated m/z value C₂₅H₂₄FN₇O₂([M+H]⁺): 474.5, measurement: 474.3.

example 49

3-tert-butyl- [1,2,4] oxadiazole-5-carboxylic acid 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzylamide

Combine { 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] in a 20mL scintillation vial]Pyrimidin-4-yl]Tert-butyl-benzyl } -carbamate (150mg, 0.355mmol, Eq: 1.00) with 3mL DCM and 3mL TFA. The solution was stirred at room temperature for 30 minutes. The solvent was removed under reduced pressure. The crude material was dissolved in DMF (4 mL). 3-tert-butyl-1, 2, 4-oxadiazole-5-carboxylic acid (66.5mg, 0.391mmol, Eq: 1.1), DIPEA (0.25mL, 1.42mmol, Eq: 4.00) and bromotripyrrolidin-1-ylphosphine were added(Pybrop) (182mg, 0.391mmol, Eq: 1.1). The reaction mixture was stirred overnight. The resulting solution was diluted with 10mL of water and 5mL of EtOAc. The solution was stirred at room temperature for a further 30 minutes. The organic phase was extracted and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure. The resulting solid was subjected to preparative HPLC (10-100% acetonitrile-water). The title compound was obtained as a yellow solid (20mg, 12% yield). LC/MS: calculated m/z value C₂₄H₂₃FN₈O₂([M+H]⁺): 475.5, measurement: 475.2.

example 50

{ 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzyl } -carbamic acid tert-butyl ester

Combine 7-benzenesulfonyl-4-chloro-6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] in 5mL of water in a 20mL sealable microwave tube]Pyrimidine (500mg, 1.34mmol, Eq: 1.00), [ 2-fluoro-4- (4, 4,5, 5-tetramethyl- [1, 3, 2-)]Dioxolane-2-yl-benzyl]Tert-butyl carbamate (541mg, 1.61mmol, Eq: 1.2) and potassium carbonate (739mg, 5.35mmol, Eq:4.00) with DME (10 mL). Adding Pd (PPh)₃)₄(375mg, 0.325mmol, Eq: 0.1). The reaction mixture was heated in a microwave at 160 ℃ for 60 minutes. The solution was washed with EtOAc and brine. The combined organic phases were dried over anhydrous sodium sulfate and the solvent was removed under reduced pressure. The crude material was purified by column chromatography (silica gel, 0-10% MeOH in DCM). The title compound was obtained as a yellow solid (150mg, 27% yield). LC/MS: calculated m/z value C₂₂H₂₃FN₆O₂([M+H]⁺): 423.4, measurement: 423.2.

example 51

N- { 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzyl } -benzamide

Combine { 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] in a 20mL scintillation vial]Pyrimidin-4-yl]-benzyl } -carbamic acid tert-butyl ester (100mg, 0.237mmol, Eq: 1.00) with 2mL DCM and 2mL TFA. The solution was stirred at room temperature for 30 minutes. The solvent was removed under reduced pressure. The crude material was dissolved in DMF (2 mL). Benzoic acid (32mg, 0.26mmol, Eq: 1.1), DIPEA (0.165mL, 0.95mmol, Eq: 4.00), and HATU (99mg, 0.26mmol, Eq: 1.1) were added. The reaction mixture was stirred overnight. The resulting solution was diluted with 10mL of water and 5mL of EtOAc. The solution was stirred at room temperature for a further 30 minutes. The organic phase was extracted and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure. The resulting solid was purified by column chromatography (silica gel, 0-10% MeOH in DCM). The title compound was obtained as a yellow solid (8mg, 8% yield). LC/MS: calculated m/z value C₂₄H₁₉FN₆O([M+H]⁺): 427.5, measurement: 427.1.

biological examples

Tyrosine kinaseEnzyme (Btk) inhibition assay

The assay is carried out by capturing radioactivity by filtration³³P phosphorylation product. Btk, biotinylated SH₂The interaction of the peptide substrate (Src homology) with ATP leads to phosphorylation of the peptide substrate. The biotinylated product is bound streptavidin agarose beads. All bound, radiolabeled products were detected by scintillation counter.

The plates assayed were 96-well polypropylene (Greiner) and 96-well 1.2 μm hydrophilic PVDF filter plates (Millipore). The concentrations reported here are the final assay concentrations: 10-100. mu.M compound, 5-10nM Btk enzyme (His-tagged, full-length), 30. mu.M peptide substrate (biotin-Aca-AAAEEIYGEI-NH) in DMSO (Burdick and Jackson)₂) 100 μ M ATP (Sigma), 8mM imidazole (Sigma, pH7.2), 8mM glycerol-2-phosphate (Sigma), 200 μ M EGTA (Roche Diagnostics), 1mM MnCl₂(Sigma), 20mM MgCl₂(Sigma), 0.1mg/ml BSA (Sigma), 2mM DTT (Sigma), 1. mu. Ci³³P ATP (Amersham), 20% streptavidin agarose beads (Amersham), 50mM EDTA (Gibco), 2M NaCl w/1% phosphoric acid (Gibco), microscint-20 (Perkin Elmer).

IC was calculated from 10 data points per compound using data generated from standard 96-well plate assay templates₅₀And (d) measuring the value. One control compound and seven unknown inhibitors were tested on each plate, twice per plate. Typically, compounds are diluted in half log (half log) starting at 100 μ M and ending at 3 nM. The control compound was staurosporine. The background was counted in the absence of peptide substrate. In the presence of peptide substrate, the total activity was determined. The following protocol was used to determine Btk inhibition.

1) Sample preparation: test compounds were added in assay buffer (imidazole, glycerol-2-phosphate, EGTA, MnCl) at half log increments₂、MgCl₂BSA).

2) Bead preparation

a.) bead rinsing by centrifugation at 500g

b.) reconstitution of the beads with PBS and EDTA to produce a 20% bead slurry

3) The reaction mixture without substrate (assay buffer, DTT, ATP, Bio-Rad antibody, etc.) was preincubated at 30 ℃,³³P ATP) and a mixture with a substrate (assay buffer, DTT, ATP,³³P ATP, peptide substrate) for 15 minutes.

4) To initiate the assay, 10 μ L of Btk in enzyme buffer (imidazole, glycerol-2-phosphate, BSA) and 10 μ L of test compound were preincubated at room temperature for 10 minutes.

5) To Btk and compound, 30 μ L of the reaction mixture containing no or substrate was added.

6) 50 μ L of the total assay mixture was incubated at 30 ℃ for 30 minutes.

7) The reaction was stopped by transferring 40. mu.L of assay to 150. mu.L of bead slurry in the filter plate.

8) After 30 minutes, the filter plate was washed with the following steps:

a.3x250μL NaCl

3 x250. mu.L NaCl containing 1% phosphoric acid

c.1x250μL H₂O

9) The plates were dried at 65 ℃ for 1 hour or at room temperature overnight.

10) Add 50. mu.L microscint-20 and count on scintillation counter³³P cpm。

Percent activity was calculated from raw data expressed as cpm

Percent activity ═ (sample-bgg)/(total activity-bgg) x100

Calculation of IC from percent Activity Using a one-site dose-response sigmoidal model₅₀

y＝A+((B-A)/(1+((x/C)^D))))

Concentration of compound (cmpd), y ═ activity, a ═ min, B ═ max, C ═ IC₅₀D1 (hill slope)

Tyrosine Kinase (BTK) inhibition of TR-FRET (time resolved FRET) assay

This BTK competitive assay uses FRET (R) ((R))the/Fluorescence Resonance energy transfer) technique measures the potency of a compound in the inactivated state of the tyrosine kinase (IC 50). The BTK-Eu complexes were incubated on ice for 1 hour, then 50nM BTK-Bioase^Tm: the starting concentration of 10nM Eu-streptavidin (Perkin-Elmer catalog # AD0062) was used. The assay buffer consisted of 20mM HEPES (pH 7.15), 0.1mM DTT, 10mM MgCl₂0.5mg/ml BSA with 3% kinase stabilizer (Fremont Biosolutions, Catalog # STB-K02). After 1h, the reaction mixture from above was diluted 10-fold with assay buffer to make 5nM BTK: 1nM Eu-streptavidin complex (donor fluorophore). 0.11nM BTK-Eu and 0.11nM kinase tracer 178(Invitrogen, Catalog # PV5593,) was then formulated into 384-well flat bottom plates (Greiner, 784076) with 18. mu.l of BTK-Eu alone as a no-negative control. Compounds tested in the assay were prepared at 10x concentration and serially diluted in DMSO at half-log increments to generate a 10 point curve. To initiate the FRET reaction, compounds made into 10 × stock solutions in DMSO were added to the plates and the plates were incubated at 14 ℃ for 18-24 h.

After incubation, the plates were read with a BMG Pherastar Fluorescent plate reader (or equivalent) and used to determine the emission energy from the europium donor fluorophore (620nm emission) and FRET (665nm emission). The negative control well values were averaged to obtain the minimum average. Positive "no inhibitor" control wells were averaged to obtain the maximum average. The maximum FRET percentage is calculated using the following equation: % max FRET 100x [ (FSR)_{Compound (I)}-FSR_{Minimum mean value})/(FSR_{Maximum mean value}-FSR_{Minimum mean value})]Wherein FSR is FRET signal ratio. The% Max FRET curve was plotted using Activity Base (Excel) and the IC50 (%), Hill slope z, and% CV were determined. Mean IC50 and standard deviation were derived from duplicate curves (singlet inhibition curves from 2 independent dilutions) using Microsoft Excel.

Representative compound data for this assay are shown in table II below.

Table II.

Compound (I)	FRET IC50(μM)	HWB IC50(μM)
			I-1	34.7
I-2	＞100
			I-3	＞100
I-4	16.3
			I-5	33.1
I-6	10.4
			I-7	16.4
I-8	＞100
			I-9	0.481
I-10	5.68
			I-11	12.2

I-12	0.121
			I-13	＞100
I-14	0.129
			I-15	1.53
I-16	0.687
			I-17	0.098	16.2
I-18	0.137
			I-19	0.137
I-20	0.062	3
			I-21	0.007	1.4
I-22	0.008	5.48
			I-23	0.007	1.4
I-24	0.065
			I-25	0.044	28.9
I-26	0.001	0.608
			I-27	0.002	0.812
I-28	0.004	2.54
			I-29	0.003	0.47
I-30	0.0007	1.04
			I-31	0.02	6.05
I-32	0.008
			I-33	0.001	0.129
I-34	0.0002	0.076
			I-35	0.008	5.5
I-36	0.004	6.74
			I-37	0.0003	0.655
I-38	0.003	3.27
			I-39	0.0002	0.028
I-40	0.0006	0.052
			I-41	0.0006	0.154
I-42	0.0005	0.23
			I-43	0.0006	0.276

I-44	0.00052	0.188
			I-45	0.0006	0.222
I-46	0.001	＞50
			I-47	0.0032	0.674
I-48	0.00019	0.076
			I-49	0.00011	0.002
I-50	1.49
			I-51	0.026

Inhibition of B cell activation in whole blood as determined by CD69 expression

The procedure for testing the ability of Btk inhibitors to inhibit B cell receptor mediated activation of B cells in human blood is as follows:

human Whole Blood (HWB) was obtained from healthy volunteers, satisfying the following limits: the preparation is not taken for 24hr, and is not suitable for smokers. Blood was collected by venipuncture into Vacutainer tubes anticoagulated with sodium heparin. Test compounds were diluted 10-fold to the desired initial drug concentration (20x) in PBS, followed by three-fold serial dilutions in 10% DMSO in PBS, resulting in a 9-point dose response curve. 5.5 μ l of each compound dilution was added in duplicate to 2mi 96 well V-bottom plates (Analytical Salesand Services, # 59623-23); to control and non-stimulated wells 5.5 μ l of 10% DMSO in PBS was added. HWB (100. mu.l) was added to each well and plates were mixed at 37C, 5% CO₂100% humidity incubation for 30 minutes. To each well (except for non-irritating wells) sheep F (ab') 2 anti-human IgM (Southern Biotech, #2022-14) (10. mu.l of 500. mu.g/ml solution, 50. mu.g/ml final concentration) was added with stirring and the plates were incubated for an additional 20 hours.

At the end of the 20 hour incubation, the samples were mixed with fluorescent probe-labeled antibodies (15. mu.l PE mouse anti-human CD20, BDPharmingen, #555623, and/or 20. mu.l APC mouse anti-human C)D69, BD Pharmingen #555533) at 37C, 5% CO₂100% humidity incubation for 30 minutes. Including induced control, unstained and single stain for compensation of adjustments and initial voltage settings. The sample was then lysed with 1ml of 1X Pharmingen Lyse Buffer (BD Pharmingen #555899), and the plate was centrifuged at 1800rpm for 5 minutes. The supernatant was removed by aspiration, the remaining pellet was re-lysed with another 1ml of 1 XPharmangen Lyse Buffer, and the plate was centrifuged (spun down) as before. The supernatant was aspirated and the remaining pellet was washed in FACs buffer (PBS + I% FBS). After the final spin, the supernatant was removed and the pellet was resuspended in 180. mu.l of FACs buffer. Samples were transferred to 96-well plates suitable for running on the HTS 96-well system of the BD LSR II flow cytometer.

Data were acquired using excitation and emission wavelengths appropriate to the fluorophore used and percentage positive Cell values were obtained using Cell quest software. The results were initially analyzed using FACS analysis software (Flow Jo). The IC50 of the test compound was defined as the concentration that reduced the percentage of CD69 positive cells by 50%, which CD69 positive cells were also CD20 positive after stimulation with anti-IgM (average of 8 control wells after subtraction of the average of 8 wells without a stimulating background). The IC50 value was calculated using XLfit software version 3, equation 201.

B-inhibition of cell activation-B cell FLIPR assay in Ramos cells

Inhibition of B cell activation by the compounds of the invention was demonstrated by determining the effect of test compounds on anti-IgM stimulated B cell responses.

The B cell FLIPR assay is a cell-based functional method to determine the effect of potential inhibitors on intracellular calcium increase caused by stimulation with anti-IgM antibodies. Ramos cells (human Burkitt's lymphoma cell line. ATCC-No. CRL-1596) were cultured in growth medium (described below). One day prior to assay, Ramos cells were resuspended in fresh growth medium (same as above) and at 0.5x10⁶The concentration of/mL was placed in a tissue culture flask. On the day of assay, cells were counted and 1 ×10⁶The concentration/mL was placed in growth medium supplemented with 1 μ M FLUO-3AM (TefLabs Cat. No. 0116, prepared in anhydrous DMSO and 10% block polyetheracid (Pluronic acid)) in tissue culture flasks and at 37 deg.C (4% CO)₂) Incubate for 1 hour. To remove extracellular dye, cells were harvested by centrifugation (5 min, 1000rpm) and washed at 1 × 10⁶cells/mL were resuspended in FLIPR buffer (described below) followed by 1 × 10⁵Cells/well were distributed in 96-well poly-D-lysine coated black/clear plates (BD catalog No. 356692). Test compounds were added at various concentrations ranging from 100 μ M to 0.03 μ M (7 concentrations, detailed below) and incubated with the cells for 30 minutes at room temperature. Ramos cells Ca were stimulated by addition of 10. mu.g/mL anti-IgM (Southern Biotech, Cat. No. 2020-01)²⁺Signals were conducted and measured on a FLIPR (Molecular Devices, using a CCD camera with an argon laser excited at 480nM to capture images of 96-well plates).

Medium/buffer:

growth medium: RPMI 1640 medium containing L-glutamine (Invitrogen, Cat. No. 61870-010), 10% fetal bovine serum (FBS, Summit Biotechnology Cat. No. FP-100-05); 1mM sodium pyruvate (Invitrogen Cat No. 11360-070).

FLIPR buffer: HBSS (Invitrogen, catalog No. 141175-079), 2mM CaCl₂(Sigma, Cat. No. C-4901), HEPES (Invitrogen, Cat. No. 15630-080), 2.5mM probenecid (Sigma, Cat. No. P-8761), 0.1% BSA (Sigma, Cat. No. A-7906), 11mM glucose (Sigma, Cat. No. G-7528).

Compound dilution details:

to obtain the highest final assay concentration of 100. mu.M, 24. mu.L of a 10mM stock solution of the compound (prepared in DMSO) was added directly to 576. mu.L of FLIPR buffer. Test compounds were diluted in FLIPR buffer (using Biomek2000 auto pipettor) to give the following dilution scheme: solvent, 1.00x10^-4M，1.00x10^-5，3.16x10^-6，1.00x10^-6，3.16x10^-7，1.00x10^-7，3.16x10^-8。

Determination and analysis：

Intracellular increase in calcium was reported using max-min statistics (using Molecular Devices FLIPR control and statistical output software, subtracting the resting baseline from the peak resulting from addition of the stimulating antibody). IC determination using non-linear curve fitting (GraphPad Prism software)₅₀。

Mouse collagen-induced arthritis (mCIA)

On day 0, mice were injected with an emulsion of type II collagen in Complete Freund's Adjuvant (CFA), injected intradermally (i.d.) at several sites at the base of the tail or on the back. Following collagen immunization, animals will develop arthritis between about 21 and 35 days. The onset of arthritis was synchronized (enhanced) by systemic administration of collagen in incomplete Freund's adjuvant (IFA; i.d.) on day 21. Animals were examined daily after day 20 for any onset of mild arthritis (score 1 or 2; described with reference to the scores below), which was an enhanced signal. After boost, mice are scored and candidate therapeutic agents are administered for a predetermined time (typically 2-3 weeks) and dosing frequency, once daily (QD) or twice daily (BID).

Rat collagen-induced arthritis (rCIA)

On day 0, rats were injected with an emulsion of type II bovine collagen in Incomplete Freund's Adjuvant (IFA), injected intradermally (i.d.) at several sites on the back. A booster injection of collagen emulsion was provided at an alternate site on the base or back of the tail on about day 7 (i.d.). Arthritis is usually observed 12-14 days after the initial collagen injection. From day 14 onwards, the progression of arthritis in the animals can be evaluated as described below (evaluation of arthritis). Animals are dosed with the candidate therapeutic agent in a prophylactic manner, starting at the time of the second challenge, and at a predetermined time (typically 2-3 weeks) and dosing frequency, once daily (QD) or twice daily (BID).

Evaluation of arthritis：

In both models, the inflammatory development of the paw and limb joints was quantified using a scoring system comprising the evaluation of 4 paws according to the criteria described below:

and (3) scoring: swelling and/or redness of the paw or toe.

Two or more joints swell.

3 ═ total swelling of the paw, involving more than two joints.

4-severe arthritis of the entire paw and toe.

The evaluation was performed as follows: baseline measurements were taken on day 0, starting again at the first sign or swelling, up to three times per week until the end of the experiment. The arthritis index was obtained for each mouse by summing the four single paw scores, with a maximum score of 16 for each animal.

Rat in vivo asthma model

Male Brown Norway (Brown-Norway) rats were sensitized i.p. once a week for three weeks (days 0, 7 and 14) with 100 μ gOA (ovalbumin) in 0.2ml alum. On day 21 (one week after the last sensitization), rats were dosed q.d. subcutaneously with solvent or compound formulation, with OA aerosol challenge (1% OA for 45 minutes) after 0.5 hours and terminated 4 or 24 hours after challenge. At sacrifice, serum and plasma were collected from all animals for serological studies and PK, respectively. The tracheal tube was inserted and the lungs were lavaged 3X with PBS. BAL fluid was analyzed for total and differential white blood cell counts. The total white blood cell count in cell aliquots (20-100. mu.l) was determined by a Coulter counter. For differential white blood cell counts, 50-200. mu.l of the sample was centrifuged in Cytospin and the slides stained with Diff-Quik. The proportion of monocytes, eosinophils, neutrophils and lymphocytes was counted under a light microscope using standard morphological criteria and expressed as a percentage. Representative inhibitors of Btk show: total leukocyte counts in BAL of OA sensitized and challenged rats were decreased compared to control levels.

The foregoing invention has been described in some detail by way of illustration and example for purposes of illustration and understanding. It will be apparent to those skilled in the art that changes and modifications may be made within the scope of the appended claims. Accordingly, it is to be understood that the above description is intended to be illustrative, and not restrictive. The scope of the invention should, therefore, be determined not with reference to the above description, but instead should be determined with reference to the following appended claims, along with the full scope of equivalents to which such claims are entitled.

All patents, patent applications, and publications cited in this application are herein incorporated by reference in their entirety for all purposes to the same extent as if each patent, patent application, or publication were individually indicated to be so incorporated by reference.

Claims

1. A compound of the formula I,

wherein:

a is phenyl;

R¹each independently is CH₂NHC(＝O)R^1’Or CH₂NHC(＝O)CH₂NHR^1’；

n is 1 or 2;

R^1”each independently is C_1-6Alkyl, halogen, cycloalkyl, heterocycloalkyl, C_1-6Alkyl heterocycloalkyl, oxo, cyano C_1-6Alkyl, hydroxy C_1-6Alkyl or C_1-6An alkoxy group;

R²is H, R³Or R⁴；

R³Is C (═ O) OR^3’、C(＝O)R^3’Or C (═ O) NH (CH)₂)₂R^3’；

R^3’Is H, C_1-6Alkyl or heterocycloalkyl;

R⁴is C_1-6Alkyl or heteroaryl, optionally substituted with one or more R^4’Substitution; and is

or a pharmaceutically acceptable salt thereof.

2. The compound of claim 1, wherein A is phenyl and R is²Is H, and n is 1.

3. The compound of claim 1, wherein R²Is H, and n is 2.

4. A compound according to claim 1 or 3, wherein n is 2, one R¹Is CH₂NHC(＝O)R^1’And R is²Is C (═ O) OR^3’、C(＝O)R^3’Or C (═ O) NH (CH)₂)₂R^3’。

5. A compound according to claim 1 or 3, whereinn is 2, one R¹Is CH₂NHC(＝O)R^1’。

6. The compound of claim 1 wherein n is 2, one R¹Is CH₂NHC(＝O)R^1’And R is²Is C_1-6Alkyl or heteroaryl.

7. A compound selected from:

4-tert-butyl-N- [4- (7H-pyrrolo [2,3-d ] pyrimidin-4-yl) -benzyl ] -benzamide;

3-chloro-N- [4- (7H-pyrrolo [2,3-d ] pyrimidin-4-yl) -benzyl ] -benzamide;

3-tert-butyl-isoxazole-5-carboxylic acid 2-fluoro-4- [6- (1-methyl-1H-pyrazol-4-yl) -7H-pyrrolo [2,3-d ] pyrimidin-4-yl ] -benzylamide;

8. Use of a compound according to any one of claims 1 to 7 in the manufacture of a medicament for the treatment of an inflammatory and/or autoimmune disorder.

9. Use of a compound according to any one of claims 1 to 7 for the manufacture of a medicament for the treatment of rheumatoid arthritis.

10. Use of a compound according to any one of claims 1 to 7 in the manufacture of a medicament for the treatment of asthma.

11. A pharmaceutical composition comprising a compound according to any one of claims 1 to 7 together with at least one pharmaceutically acceptable carrier, excipient or diluent.