RU2830171C1 - Btk inhibitors - Google Patents

Abstract

FIELD: pharmaceutics.

SUBSTANCE: invention relates to compounds of formula (I)

where R¹ is C_1-6alkyl, where R¹ can be substituted with one R⁵; k equals 0; R³ is halogen; n is 0–2; R⁴ is halogen; m is 0–2; A is =N- or =C(R⁶)-; R⁵ is hydroxy; R⁶ is hydrogen; or a pharmaceutically acceptable salt thereof, as well as a pharmaceutical composition and a method of treating diseases or disorders mediated in whole or in part by BTK, and a method of inhibiting BTK.

EFFECT: novel compounds are obtained, which have inhibitory activity on BTK, where BTK is a wild-type BTK or BTK with a C481 mutation.

17 cl, 15 ex

Description

FIELD OF INVENTION

The present application relates to ocean-substituted imidazopyrazine and imidazotriazine Bruton tyrosine kinase (BTK) inhibitors, including mutant BTK, useful in the treatment of BTK-related diseases or disorders. These compounds have potential use in the treatment of immune disorders, cancer, cardiovascular diseases, viral infections, inflammation, metabolic/endocrine disorders, and neurological disorders.

In particular, the application relates to compounds and compositions thereof that inhibit VTK, methods of treating diseases or disorders associated with VTK, and methods of synthesizing these compounds.

PRIOR ART

Bruton tyrosine kinase (BTK), also known as tyrosine-protein kinase BTK, is a member of the Tes family of tyrosine kinases and plays an important role in the regulation of early B cell development and the activation and survival of mature B cells (Hunter, Cell, 87, 50, 823-829). The BTK enzyme is encoded by the BTK gene and has been shown to initiate a number of cellular processes including cell proliferation, survival, differentiation, motility, angiogenesis, cytokine production, and antigen presentation.

In BTK-deficient mouse models, BTK has been shown to play a role in allergic diseases and/or autoimmune diseases and/or inflammatory diseases; and BTK inhibition has potential benefit in the treatment of diseases such as systemic lupus erythematosus (SLE), urticaria/Sjogren's syndrome, rheumatoid arthritis, vasculitis, idiopathic thrombocytopenic purpura (ITP), myasthenia gravis, allergic rhinitis, and asthma.

The role of BTK in apoptosis also suggests that inhibiting BTK activity may be useful in treating cancers such as B-cell lymphoma, leukemia, and other hematological malignancies. Additionally, BTK plays a role in osteoclast function, so inhibiting BTK activity may have potential benefits in treating bone diseases such as osteoporosis.

Approved compounds that inhibit BTK include ibrutinib (B-cell neoplasms such as mantle cell lymphoma, chronic lymphocytic leukemia (CLL), and Waldenstrom's macroglobulinemia); acalabrutinib (mantle cell lymphoma and CLL); and zanubrutinib (mantle cell lymphoma). In addition, several BTK inhibitors are in clinical trials, including evobrutinib (multiple sclerosis); ABBV-105 (systemic lupus erythematosus (SLE)); ONO-4059/GS-4059 (non-Hodgkin lymphoma and CLL); spebrutinib (relapsed or refractory B-cell non-Hodgkin lymphoma, CLL, and Waldenstrom's macroglobulinemia); and HM71224 (autoimmune diseases).

Despite significant therapeutic advances in the treatment of B-cell neoplasms using BTK inhibitors, cases of primary and secondary resistance have emerged with unfavorable outcomes and limited treatment options.

Covalent (irreversible) BTK inhibitors such as ibrutinib and acalabrutinib bind to the C481 site of BTK, rendering it inactive for the kinase. This binding is permanent until the BTK protein is degraded. The advantage of these irreversible inhibitors is that they are potent and usually only effective for a short period of time. However, their clinical utility is limited by off-target toxicity, resulting in high rates of treatment discontinuation and acquired resistance due to BTK C481 mutations that disrupt covalent binding to BTK, reducing the binding affinity of the compounds and decreasing their ability to inhibit BTK enzymatic activity (Leukaemia 2015, Apr; 29(4):895–900). The majority (more than 50%) of CLL patients who progress with covalent BTK inhibitors become resistant to treatment due to the development of the C481S mutation (N. Engl. J. Med. 370; 24, 2014; JAMA Oncol. 2015; 1(1):80–87 and J. Clin. Oncol. 35:1437–1443, 2017).

Primary central nervous system lymphoma (PCNSL) is a disease in which malignant (cancerous) cells originate in the lymphatic tissue of the brain and/or spinal cord, and accounts for approximately 1% of all lymphomas and 2% to 5% of all primary brain tumors. The vast majority (approximately 95%) of PCNSL are diffuse large B-cell lymphomas (DLBCL). Mutations in the CD79B and MYD88 genes are common (approximately 30-80%) in PCNSL (Neuropathol. Appl. Neurobiol. 2016 Apr; 42(3):279-90). Although BTK inhibitors have not been approved for the treatment of DLBCL to date, data suggest that DLBCL with CD79B and MYD88 mutations are more sensitive to BTK inhibition (Nat. Med. 2015 Aug; 21(8):922-6).

Secondary CNS lymphoma (SCNSL) refers to dissemination to the central nervous system of lymphoma arising elsewhere (as opposed to primary CNS lymphoma). It is usually non-Hodgkin lymphoma and may present as an isolated relapse or may be part of a systemic illness at presentation. Unlike primary CNS lymphoma, it more commonly involves the leptomeninges.

PRN2246 (SAR442168), a blood-brain barrier (BBB)-permeable covalent inhibitor of VTK, was well tolerated in a phase I multiple sclerosis (MS) trial. In addition, some trials (Grommes C, et al., Cancer Discov. 2017 Sep;7(9): 1018–1029; Grommes C, et al., Blood. 2019; 133(5):436–445; Lionakis et al., 2017, Cancer Cell 31, 833–843; and Soussain C et al., Eur J Cancer. 2019 Aug; 117:121–130) suggested that high-dose ibrutinib (840 mg) would be effective in CNS lymphomas (both PCNSL and secondary lymphoma of the nervous system (SCLNS)), but to date, no BTK inhibitors targeting the BTK-C481 mutation or with activity in PCNSL have been approved, leaving both unmet medical needs.

WO 2009/143051 discloses certain substituted imidazopyrazines and imidazotriazines, including certain hexane-substituted imidazopyrazines and imidazotriazines, as inhibitors of activated p21cdc42Hs-associated kinase (ASK1). However, the compounds of WO 2009/143051 exhibit high clearance from human hepatocytes, meaning that these compounds potentially fail to provide sufficient sustained drug action even at the maximum absorbable dose (rendering the compounds ineffective); and/or very high doses are required to inhibit the target, resulting in a high maximum drug concentration (potentially resulting in secondary pharmacological (i.e. adverse) effects and toxicity issues).

WO 2017111787A1 discloses a tetrahydropyranyl amino pyrrolopyrimidinone that modulates BTK activity; WO 2018039310A1 discloses amino pyrrolopyrimidinone compounds and methods of using them; WO 2017103611A1 discloses compounds useful as BTK inhibitors; and WO 2011152351 discloses purinone derivatives having selective BTK inhibitory activity. However, none of these compounds have the combination of desirable properties that the compounds of the present invention have.

Here, some novel oceanic substituted imidazopyrazines and imidazotriazines are disclosed that are potent, selective inhibitors of both wild-type and C481-mutated BTK (e.g., C481S, C481Y, C481R, or C481F). These compounds are non-covalent, reversible inhibitors, exhibit low clearance from human hepatocytes, and penetrate the blood-brain barrier (BBB).

SUMMARY OF THE INVENTION

Compounds of formula (I) are disclosed herein:

and their pharmaceutically acceptable salts and their use as VTK inhibitors, in particular in therapy.

DETAILED DESCRIPTION OF THE INVENTION

Many embodiments of the invention are described in detail in the description and will be obvious to those skilled in the art. The invention should not be interpreted as limited to any of the embodiments given, and the claims are embodiments. It should be understood that some features of the present invention, which for clarity are described in the context of separate embodiments, can also be presented in combination in a single embodiment. Conversely, various features of the present invention, which for brevity are described in the context of a single embodiment, can also be presented separately or in any suitable subcombination.

Herein is disclosed a compound of formula (I):

Where:

R ¹ is selected from hydrogen, C _1-6 alkyl, C _1-6 alkoxy, N-C _1-6 alkylamino, N,N-(C _1-6 alkyl) ₂ amino, carbocyclyl and heterocyclyl; wherein R ¹ may be optionally substituted with one or more R ⁵ ;

R ² is selected from halogen, C _1-3 alkyl, C _1-3 alkoxy, carbocyclyl and heterocyclyl; or two R ² , either on the same atom or on adjacent atoms, may, together with the atoms to which they are attached, form a 3-7-membered ring;

k is 0-4;

R ³ is selected from halogen, C _1-3 alkyl and C _1-3 alkoxy;

n is 0-4;

R ⁴ is selected from halogen, C _1-3 alkyl and C _1-3 alkoxy;

m is 0-5;

A represents =N- or =C(R ⁶ )-;

R ⁵ is selected from halogen, hydroxy, C _1-6 alkoxy, amino, N-C _1-6 alkylamino, N,N-(C _1-6 alkyl) ₂ amino, carbocyclyl and heterocyclyl; wherein R ⁵ may be independently optionally substituted with one or more R ⁷ ;

R ⁶ is selected from hydrogen and halogen;

R ⁷ is selected from halogen, hydroxy, amino, C _1-3 alkyl and C _1-3 alkoxy;

or a pharmaceutically acceptable salt thereof.

In one embodiment R ¹ is selected from hydrogen and C _1-6 alkyl; wherein R ¹ may be optionally substituted with one R ⁵ ; wherein R ⁵ is selected from hydroxy, C _1-6 alkoxy, N,N-(C _1-6 alkyl) ₂ amino and heterocyclyl.

In one embodiment R ¹ is selected from hydrogen and C _1-3 alkyl; wherein R ¹ may be optionally substituted with one R ⁵ ; wherein R ⁵ is selected from hydroxy, C _1-3 alkoxy, N,N-(C _1-2 alkyl) ₂ amino and azetidinyl.

In one embodiment R ¹ is selected from hydrogen, methyl, hydroxymethyl, methoxymethyl, N,N-dimethylaminomethyl and azetidin-1-ylmethyl.

In one embodiment, R ¹ is hydroxymethyl.

In one embodiment R ² is selected from halogen or C _1-3 alkoxy.

In one embodiment, R ² is selected from fluoro or methoxy.

In one embodiment, two or more R ² , either on the same atom or on adjacent atoms, may, together with the atoms to which they are attached, form a 3- to 7-membered ring.

In one embodiment, two or more R ² on a single atom may, together with the atom to which they are attached, form a 3- to 7-membered ring.

In one embodiment, two or more R ^2s on adjacent atoms may, together with the atoms to which they are attached, form a 3- to 7-membered ring.

In one embodiment, k is equal to 0.

In one embodiment, k is equal to 1.

In one embodiment, k is equal to 2.

In one embodiment, k is equal to 3.

In one embodiment, k is 4.

In one embodiment, R ³ is halogen.

In one embodiment, R ³ is fluoro.

In one embodiment n is 0-2.

In one embodiment n is 0.

In one embodiment n is equal to 1.

In one embodiment n is equal to 2.

In one embodiment n is equal to 3.

In one embodiment n is 4.

In one embodiment, R ⁴ is halogen.

In one embodiment, R ⁴ is fluoro.

In one embodiment m is 0-2.

In one embodiment m is equal to 0.

In one embodiment m is equal to 1.

In one embodiment m is equal to 2.

In one embodiment m is equal to 3.

In one embodiment m is equal to 4.

In one incarnation m is equal to 5.

In one embodiment, A is =N- or =C(H)-.

In one embodiment, A is =N-.

In one embodiment, A is =C(R ⁶ )-.

In one embodiment, A is =C(H)-.

The compound of formula (I) (where R ¹ is not hydrogen) contains two chiral centers (marked with “*”):

These chiral centers can exist in the "trans" configuration (meaning that the two substituents on the oxane ring are located on opposite sides of the oxane ring); and the "cis" configuration (meaning that the two substituents on the oxane ring are located on the same side of the oxane ring). Structures (IA) and (IB) below represent the cis isomers of compounds of formula (I), structures (IC) and (ID) below represent the trans isomers of compounds of formula (I).

In one aspect of the invention, the compound of formula (I) is a trans compound of formula (I).

In one aspect of the invention, the compound of formula (I) is a cis-compound of formula (I).

In one aspect of the invention, the compound of formula (I) is a compound of formula (IA):

In one aspect of the invention, the compound of formula (I) is a compound of formula (IB):

In one aspect of the invention, the compound of formula (I) is a compound of formula (IC):

In one aspect of the invention, the compound of formula (I) is a compound of formula (ID):

In one aspect of the invention, the compound of formula (I) is selected from

(5-(8-amino-1-(4-phenoxyphenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol;

(5-(8-amino-1-(2-fluoro-4-phenoxyphenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol;

(5-(8-amino-1-(2,3-difluoro-4-phenoxyphenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol;

(5-(8-amino-1-(4-(2,3-difluorophenoxy)phenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol;

(5-(4-amino-5-(2-fluoro-4-phenoxyphenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-yl)methanol;

(5-(4-amino-5-(2-fluoro-4-phenoxyphenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-yl)methanol;

(5-(4-amino-5-(2,3-difluoro-4-phenoxyphenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-yl)methanol;

(5-(4-amino-5-(4-(2,3-Difluorophenoxy)phenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-yl)methanol;

3-(6-((dimethylamino)methyl)tetrahydro-2H-pyran-3-yl)-1-(2-fluoro-4-phenoxyphenyl)imidazo[1,5-a]pyrazin-8-amine;

7-(6-((dimethylamino)methyl)tetrahydro-2H-pyran-3-yl)-5-(2-fluoro-4-phenoxyphenyl)imidazo[5,1-f][1,2,4]triazin-4-amine;

7-(6-(azetidin-1-ylmethyl)tetrahydro-2H-pyran-3-yl)-5-(2-fluoro-4-phenoxyphenyl)imidazo[51,-f][1,2,4]triazin-4-amine;

5-(2-fluoro-4-phenoxyphenyl)-7-(6-(methoxymethyl)tetrahydro-2H-pyran-3-yl)imidazo[5,1-f][1,2,4]triazin-4-amine;

5-(2-fluoro-4-phenoxyphenyl)-7-(tetrahydro-2H-pyran-3-yl)imidazo[5,1-f][1,2,4]triazin-4-amine; and

5-(2-fluoro-4-phenoxyphenyl)-7-(6-methyltetrahydro-2H-pyran-3-yl)imidazo[5,1-f][1,2,4]triazin-4-amine.

In one aspect of the invention, the compound of formula (I) is selected from

(5-(8-amino-1-(4-phenoxyphenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol;

(5-(8-amino-1-(2-fluoro-4-phenoxyphenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol;

(5-(8-amino-1-(2,3-difluoro-4-phenoxyphenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol;

(5-(8-amino-1-(4-(2,3-difluorophenoxy)phenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol;

(5-(4-amino-5-(2-fluoro-4-phenoxyphenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-yl)methanol;

(5-(4-amino-5-(2-fluoro-4-phenoxyphenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-yl)methanol;

(5-(4-amino-5-(2,3-difluoro-4-phenoxyphenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-yl)methanol;

(5-(4-amino-5-(4-(2,3-difluorophenoxy)phenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-yl)methanol;

3-(6-((dimethylamino)methyl)tetrahydro-2H-pyran-3-yl)-1-(2-fluoro-4-phenoxyphenyl)imidazo[1,5-a]pyrazin-8-amine;

7-(6-((dimethylamino)methyl)tetrahydro-2H-pyran-3-yl)-5-(2-fluoro-4-phenoxyphenyl)imidazo[5,1-f][1,2,4]triazin-4-amine;

7-(6-(azetidin-1-ylmethyl)tetrahydro-2H-pyran-3-yl)-5-(2-fluoro-4-phenoxyphenyl)imidazo[5,1-f][1,2,4]triazin-4-amine;

5-(2-fluoro-4-phenoxyphenyl)-7-(6-(methoxymethyl)tetrahydro-2H-pyran-3-yl)imidazo[5,1-f][1,2,4]triazin-4-amine;

5-(2-fluoro-4-phenoxyphenyl)-7-(tetrahydro-2H-pyran-3-yl)imidazo[5,1-f][1,2,4]triazin-4-amine; and

5-(2-fluoro-4-phenoxyphenyl)-7-(6-methyltetrahydro-2H-pyran-3-yl)imidazo[5,1-f][1,2,4]triazin-4-amine;

or a pharmaceutically acceptable salt thereof.

In one aspect of the invention, the compound of formula (I) is selected from

((2R,5R)-5-(8-amino-1-(4-phenoxyphenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol;

((2R,5R)-5-(8-amino-1-(2-fluoro-4-phenoxyphenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol;

((2R,5R)-5-(8-amino-1-(2,3-difluoro-4-phenoxyphenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol;

((2R,5R)-5-(8-amino-1-(4-(2,3-difluorophenoxy)phenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol;

((2R,5R)-5-(4-amino-5-(2-fluoro-4-phenoxyphenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-yl)methanol;

((2R,5R)-5-(4-amino-5-(2-fluoro-4-phenoxyphenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-yl)methanol;

((2R,5R)-5-(4-amino-5-(2,3-difluoro-4-phenoxyphenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-yl)methanol;

((2R,5R)-5-(4-amino-5-(4-(2,3-Difluorophenoxy)phenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-yl)methanol;

3-((3R,6R)-6-((dimethylamino)methyl)tetrahydro-2H-pyran-3-yl)-1-(2-fluoro-4-phenoxyphenyl)imidazo[1,5-a]pyrazin-8-amine;

7-((3R,6R)-6-((dimethylamino)methyl)tetrahydro-2H-pyran-3-yl)-5-(2-fluoro-4-phenoxyphenyl)imidazo[5,1-f][1,2,4]triazin-4-amine;

7-((3R,6R)-6-(azetidin-l-ylmethyl)tetrahydro-2H-pyran-3-yl)-5-(2-fluoro-4-phenoxyphenyl)imidazo[5,1-f],[ 1,2,4]triazin-4-amine;

5-(2-fluoro-4-phenoxyphenyl)-7-((3R,6R)-6-(methoxymethyl)tetrahydro-2H-pyran-3-yl)imidazo[5,1-f][1,2,4]triazin-4-amine;

(R)-5-(2-fluoro-4-phenoxyphenyl)-7-(tetrahydro-2H-pyran-3-yl)imidazo[5,1-f][1,2,4]triazin-4-amine; And

5-(2-fluoro-4-phenoxyphenyl)-7-((3R,6R)-6-methyltetrahydro-2H-pyran-3-yl)imidazo[5,1-f][1,2,4]triazin-4-amine;

or a pharmaceutically acceptable salt thereof.

In one aspect of the invention, the compound of formula (I) is selected from

((2S,5S)-5-(8-amino-1-(4-phenoxyphenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol;

((2S,5S)-5-(8-amino-1-(2-fluoro-4-phenoxyphenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol;

((2S,5S)-5-(8-amino-1-(2,3-difluoro-4-phenoxyphenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol;

((2S,5S)-5-(8-amino-1-(4-(2,3-Difluorophenoxy)phenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol;

((2S,5S)-5-(4-amino-5-(2-fluoro-4-phenoxyphenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-yl)methanol;

((2S,5S)-5-(4-amino-5-(2-fluoro-4-phenoxyphenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-yl)methanol;

((2S,5S)-5-(4-amino-5-(2,3-difluoro-4-phenoxyphenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-yl)methanol;

((2S,5S)-5-(4-amino-5-(4-(2,3-difluorophenoxy)phenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-yl)methanol;

3-((3S,6S)-6-((dimethylamino)methyl)tetrahydro-2H-pyran-3-yl)-1-(2-fluoro-4-phenoxyphenyl)imidazo[1,5-a]pyrazin-8-amine;

7-((3S,6S)-6-((dimethylamino)methyl)tetrahydro-2H-pyran-3-yl)-5-(2-fluoro-4-phenoxyphenyl)imidazo[5,1-f][1,2,4]triazin-4-amine;

7-((3S,6S)-6-(azetidin-1-ylmethyl)tetrahydro-2H-pyran-3-yl)-5-(2-fluoro-4-phenoxyphenyl)imidazo[5,1-f][1,2,4]triazin-4-amine;

5-(2-fluoro-4-phenoxyphenyl)-7-((3S,6S)-6-(methoxymethyl)tetrahydro-2H-pyran-3-yl)imidazo[5,1-f][1,2,4]triazin-4-amine;

(S)-5-(2-fluoro-4-phenoxyphenyl)-7-(tetrahydro-2H-pyran-3-yl)imidazo[5,1-f][1,2,4]triazin-4-amine; and

5-(2-fluoro-4-phenoxyphenyl)-7-((3S,6S)-6-methyltetrahydro-2H-pyran-3-yl)imidazo[5,1-f][1,2,4]triazin- 4-amine;

or a pharmaceutically acceptable salt thereof.

In one aspect of the invention, the compound of formula (I) is selected from ((2S,5R)-5-(8-amino-1-(4-phenoxyphenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol;

((2S,5R)-5-(8-amino-1-(2-fluoro-4-phenoxyphenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol;

((2S,5R)-5-(8-amino-1-(2,3-difluoro-4-phenoxyphenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol;

((2S,5R)-5-(8-amino-1-(4-(2,3-difluorophenoxy)phenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol;

((2S,5R)-5-(4-amino-5-(2-fluoro-4-phenoxyphenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-yl)methanol;

((2S,5R)-5-(4-amino-5-(2-fluoro-4-phenoxyphenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-yl)methanol;

((2S,5R)-5-(4-amino-5-(2,3-difluoro-4-phenoxyphenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-yl)methanol;

((2S,5R)-5-(4-amino-5-(4-(2,3-Difluorophenoxy)phenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-yl)methanol;

3-((3S,6R)-6-((dimethylamino)methyl)tetrahydro-2H-pyran-3-yl)-1-(2-fluoro-4-phenoxyphenyl)imidazo[1,5-a]pyrazin-8-amine;

7-((3S,6R)-6-((dimethylamino)methyl)tetrahydro-2H-pyran-3-yl)-5-(2-fluoro-4-phenoxyphenyl)imidazo[5,1-f][1,2,4]triazin-4-amine;

7-((3S,6R)-6-(azetidin-1-ylmethyl)tetrahydro-2H-pyran-3-yl)-5-(2-fluoro-4-phenoxyphenyl)imidazo[5,1-f],[1,2,4]triazin-4-amine;

5-(2-fluoro-4-phenoxyphenyl)-7-((3S,6R)-6-(methoxymethyl)tetrahydro-2H-pyran-3-yl)imidazo[5,1-f][1,2,4]triazin-4-amine; and

5-(2-fluoro-4-phenoxyphenyl)-7-((3S,6R)-6-methyltetrahydro-2H-pyran-3-yl)imidazo[5,1-f][1,2,4]triazin-4-amine;

or a pharmaceutically acceptable salt thereof.

In one aspect of the invention, the compound of formula (I) is selected from ((2R,5S)-5-(8-amino-1-(4-phenoxyphenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol;

((2R,5S)-5-(8-amino-1-(2-fluoro-4-phenoxyphenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol;

((2R,5S)-5-(8-amino-1-(2,3-difluoro-4-phenoxyphenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol;

((2R,5S)-5-(8-amino-1-(4-(2,3-Difluorophenoxy)phenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol;

((2R,5S)-5-(4-amino-5-(2-fluoro-4-phenoxyphenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-yl)methanol;

((2R,5S)-5-(4-amino-5-(2-fluoro-4-phenoxyphenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-yl)methanol;

((2R,5S)-5-(4-amino-5-(2,3-difluoro-4-phenoxyphenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-yl)methanol;

((2R,5S)-5-(4-amino-5-(4-(2,3-difluorophenoxy)phenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-yl)methanol;

3-((3R,6S)-6-((dimethylamino)methyl)tetrahydro-2H-pyran-3-yl)-1-(2-fluoro-4-phenoxyphenyl)imidazo[1,5-a]pyrazin-8-amine;

7-((3R,6S)-6-((dimethylamino)methyl)tetrahydro-2H-pyran-3-yl)-5-(2-fluoro-4-phenoxyphenyl)imidazo[5,1-f][1,2,4]triazin-4-amine;

7-((3R,6S)-6-(azetidin-1-ylmethyl)tetrahydro-2H-pyran-3-yl)-5-(2-fluoro-4-phenoxyphenyl)imidazo[5,1-f][1,2,4]triazin-4-amine;

5-(2-fluoro-4-phenoxyphenyl)-7-((3R,6S)-6-(methoxymethyl)tetrahydro-2H-pyran-3-yl)imidazo[5,1-f][1,2,4]triazin-4-amine; and

5-(2-fluoro-4-phenoxyphenyl)-7-((3R,6S)-6-methyltetrahydro-2H-pyran-3-yl)imidazo[5,1-f][1,2,4]triazin-4-amine;

or a pharmaceutically acceptable salt thereof.

In one aspect, the invention provides any compound of formula (I) disclosed herein.

In one aspect, the invention provides any compound of formula (I) disclosed herein, or a pharmaceutically acceptable salt thereof.

In one aspect of the invention there is provided a synthetic intermediate used to prepare a compound of formula (I) as disclosed herein.

In one aspect of the invention there is provided a synthetic intermediate used to prepare a compound of formula (I) as disclosed herein, or a pharmaceutically acceptable salt thereof.

Linking substituents are described in various parts of the present invention. In cases where a structure clearly requires a linking group, then the Markush variables listed for that group are intended to be linking groups. For example, if a structure requires a linking group and the Markush group definition lists "alkyl" for that variable, then the "alkyl" is intended to be a linking alkylene group.

As used herein, the term "substituted" when referring to a chemical group means that the chemical group has one or more hydrogen atoms that have been removed and replaced by substituents. As used herein, the term "substituent" has the ordinary meaning known in the art and refers to a chemical moiety that is covalently attached or, where appropriate, fused to the parent group. As used herein, the term "optionally substituted" or "optionally... substituted" means that the chemical group may have no substituents (i.e., be unsubstituted) or may have one or more substituents (i.e., be substituted). It should be understood that substitution at a given atom is limited by valence.

The term "C _ij " as used herein denotes a range of carbon atom numbers, where i and j are integers, and the range of carbon atom numbers includes the finite numbers (i.e. i and j) and every integer between them, and where j is greater than i. For example, C _1-6 denotes a range of one to six carbon atoms, including one carbon atom, two carbon atoms, three carbon atoms, four carbon atoms, five carbon atoms and six carbon atoms. In some embodiments, the term "C _1-6 " denotes from 1 to 6, in particular from 1 to 5, in particular from 1 to 4, in particular from 1 to 3 or in particular from 1 to 2 carbon atoms.

The term "alkyl" as used herein, whether used as part of another term or by itself, refers to a saturated hydrocarbon chain. The hydrocarbon chain referred to above may be linear or branched. The term "C _ij alkyl" refers to an alkyl having from i to j carbon atoms. Examples of C _1-6 alkyl include methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl; higher homologues such as 2-methyl-1-butyl, n-pentyl, 3-pentyl, n-hexyl, 1,2,2-trimethylpropyl and the like, but are not limited thereto. Examples of the "C _1-3 alkyl" group include methyl, ethyl, propyl and isopropyl.

As used herein, the term "halogen" refers to an atom selected from fluorine, chlorine, bromine, and iodine.

As used herein, the term "alkoxy", whether used as part of another term or by itself, refers to a group of the formula -O-alkyl. The term "C _ij alkoxy" means that the alkyl moiety of the alkoxy group has from i to j carbon atoms. Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy (e.g., n-propoxy and isopropoxy), tert-butoxy, and the like. Examples of the "O-valkoxy" group are methoxy, ethoxy, and propoxy. Examples of the "C _1-3 alkoxy" group are methoxy, ethoxy, and propoxy.

Examples of the "N-(C _1-6 alkyl)amino" group are methylamino and ethylamino. Examples of the "N,N-(C _1-6 alkyl) ₂ amino" group are N,N-dimethylamino, N,N-diethylamino, and N-ethyl-N-methylamino.

As used herein, the term "carbocyclyl", whether used as part of another term or by itself, refers to a saturated monocyclic ring in which all ring atoms are carbon and which contains at least three carbon atoms forming a ring. In some embodiments, a carbocyclyl may contain from 3 to 7 ring carbon atoms, or from 3 to 6 ring carbon atoms. In some embodiments, the _-CH2- group of the ring may be replaced by a -C(O)- group of the ring. Examples of carbocyclyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl.

As used herein, the term "heterocyclyl" refers to a monocyclic, saturated carbocyclyl group wherein one or more (e.g., 1, 2, or 3) ring atoms are replaced with heteroatoms that include, but are not limited to, oxygen, sulfur, nitrogen, phosphorus, and the like. In some embodiments, the _-CH2- group of the ring may be replaced with a -C(O)- group of the ring. In some embodiments, the sulfur ring atom may be optionally oxidized to form S-oxides. In some embodiments, the heterocyclyl is attached through carbon. In some embodiments, the heterocyclyl is attached through nitrogen. Examples of heterocyclyl groups include, but are not limited to, azetidinyl, piperidyl, pyrrolidinyl, tetrahydrofuryl, piperidinyl, piperazinyl, morpholinyl, and the like.

In one embodiment, two R ² on the same atom, together with the atom to which they are attached, form a 3-7 membered ring. The resulting "spiro rings" have two rings (one of which is an ocean of formula (I)) connected through a single common atom. The neoxane ring may be a 3-7 membered carbocyclyl ring or a 3-7 membered heterocyclic ring. Examples of two R ² on the same atom, together with the atom to which they are attached, forming a 3-7 membered ring (shown with an ocean of formula (I)), include:

(Where shows attachment to the rest of the molecule).

In one embodiment, two R ² on adjacent atoms, together with the atoms to which they are attached, form a 3- to 7-membered ring. The resulting "fused rings" have two rings (one of which is an ocean of formula (I)) sharing two adjacent atoms. The neoxane ring may be a 3- to 7-membered carbocyclyl ring or a 3- to 7-membered heterocyclic ring. Examples of two R ² on adjacent atoms, together with the atoms to which they are attached, forming a 3- to 7-membered ring (shown with an ocean of formula (I)), include:

(Where shows attachment to the rest of the molecule).

A "compound" according to the present invention is meant to include all stereoisomers, geometric isomers and tautomers of the structures shown unless otherwise specified.

The term "stereoisomer" refers to any of the various stereoisomeric configurations (e.g., enantiomers, diastereomers, and racemates) of an asymmetric compound (e.g., those having one or more asymmetrically substituted carbon atoms, or "asymmetric centers"). Compounds of the present invention that contain asymmetric centers may be isolated in optically active (enantiomers or diastereomers) or optically inactive (racemic) forms. The term "enantiomer" includes pairs of stereoisomers that are non-superimposable mirror images of each other. A 1:1 mixture of a pair of enantiomers is a "racemic mixture." The terms "diastereomers" or "diastereoisomers" include stereoisomers that have at least two asymmetric atoms but are not mirror images of each other. Certain compounds containing one or more asymmetric centers may give rise to enantiomers, diastereomers, or other stereoisomeric forms which may be designated by the absolute configuration as (R)- or (S)- at each asymmetric center according to the R-S Cahn-Ingold-Prelog system. Separated compounds whose absolute configuration is unknown may be designated by using the term "or" at the asymmetric center. Methods for preparing optically active forms from racemic mixtures, such as separation by HPLC (high performance liquid chromatography) or stereoselective synthesis, are known in the art.

The terms "geometric isomers" or "cis and trans isomers" refer to compounds with the same formula but whose functional groups are rotated in a different orientation in three-dimensional space.

The term "tautomers" includes prototropic tautomers, which are isomeric states of protonation of compounds having the same formula and overall charge. Examples of prototropic tautomers include ketone-enol pairs, amide-imidic acid pairs, lactam-lactim pairs, enamine-imine pairs, and ring forms in which a proton can occupy two or more positions of a heterocyclic system, such as, but not limited to, 1H- and 3H-imidazole, 1H-, 2H-, and 4H-1,2,4-triazole, 1H- and 2H-isoindole, and 1H- and 2H-pyrazole. Tautomers may exist in equilibrium or be sterically locked into one form by appropriate substitution. Compounds of the present invention identified by name or structure as one particular tautomeric form are intended to include other tautomeric forms unless otherwise indicated.

It is also intended that a "compound" according to the present invention also includes all isotopes of the atoms in the compounds. Isotopes of an atom include atoms having the same atomic number but different mass numbers. For example, unless otherwise stated, hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine, bromine, or iodine in a "compound" of the present invention are also intended to include, but are not limited to, isotopes such as: ¹ H, ² H, ³ H, ¹¹ C, ¹² C, ¹³ C, ¹⁴ C, ¹⁴ N, ¹⁵ N, ¹⁶ O, ¹⁷ O, ¹⁸ O, ³¹ P, ³² P, ³² S, ³³ S, ³⁴ S, ³⁶ S, ¹⁷ F, ¹⁹ F, ³⁵ Cl, ³⁷ Cl, ⁷⁹ Br, ⁸¹ Br, ¹²⁷ I, and ¹³¹ I. In some embodiments, hydrogen includes protium, deuterium, and tritium. In some embodiments, the term "deuterium-substituted" or "deuterium-substituted" means replacing another isoform of hydrogen (e.g., protium) in a chemical group with deuterium. In some embodiments, carbon includes ^12C and ^13C . In some embodiments, a "compound" of the present invention includes only isotopes of hydrogen in the compound. In some embodiments, a "compound" of the present invention includes only naturally occurring isotopes of atoms.

It is also to be understood that the "compound" of the present invention may exist in solvated as well as unsolvated forms, such as, for example, hydrated forms, solid forms, and the present invention is intended to include all such solvated and unsolvated forms.

It is further understood that the "compound" of the present invention may exist in the form of pharmaceutically acceptable salts.

As used herein, the term "pharmaceutically acceptable" refers to those compounds, substances, compositions, and/or dosage forms that are suitable, within the scope of sound medical judgment, for use in contact with human and animal tissues without undue toxicity, irritation, allergic reactions, or other problems or complications, commensurate with a reasonable benefit/risk ratio. In some embodiments, compounds, substances, compositions, and/or dosage forms that are pharmaceutically acceptable are those that are approved by a regulatory agency (such as the U.S. Food and Drug Administration, the China Food and Drug Administration, or the European Medicines Agency) or listed in a recognized pharmacopeia (such as the United States Pharmacopoeia, the Chinese Pharmacopoeia, or the European Pharmacopoeia) for use in animals, and more particularly in humans.

The term "pharmaceutically acceptable salts" as used herein refers to derivatives of the compounds of the present invention in which the parent compound is modified by converting an existing acidic moiety (e.g., carboxyl, etc.) or a basic moiety (e.g., amine, alkali, etc.) into its salt form. In many cases, the compounds of the present invention are capable of forming acidic and/or basic salts due to the presence of amino and/or carboxyl groups or similar groups. And pharmaceutically acceptable salts are acid and/or base salts that retain the biological effectiveness and properties of the parent compound that are generally not undesirable from a biological or other point of view. Suitable pharmaceutically acceptable salts of the compound of the present invention include, for example, an acid addition salt which can be formed, for example, from an inorganic acid (e.g. hydrochloric, hydrobromic, sulfuric, nitric, phosphoric acid and the like) or an organic acid (e.g. formic, acetic, propionic, glycolic, oxalic, maleic, malonic, succinic, fumaric, tartaric, trimesic, citric, lactic, phenylacetic, benzoic, mandelic, methanesulfonic, naphthalenedisulfonic, ethanesulfonic, toluenesulfonic, trifluoroacetic, salicylic, sulfosalicylic acids and the like).

Suitable pharmaceutically acceptable salts of the compound of the present invention also include, for example, a base addition salt, which can be formed, for example, from inorganic bases (e.g., sodium, potassium, ammonium salts and hydroxides, carbonates, bicarbonates of metals from Groups I to XII of the periodic table, such as calcium, magnesium, iron, silver, zinc, copper and the like) or organic bases (e.g., primary, secondary and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, basic ion exchange resins and the like). Some organic amines include, but are not limited to, isopropylamine, benzathine, cholinate, diethanolamine, diethylamine, lysine, meglumine, piperazine and tromethamine. One skilled in the art will understand that it is also possible to add acids or bases to form acid/base addition salts other than those shown in the examples. Lists of additional suitable salts can be found, for example, in "Remington's Pharmaceutical Sciences", 20th ed., Mack Publishing Company, Easton, Pa., (1985); and in "Handbook of Pharmaceutical Salts: Properties, Selection, and Use", Stahl and Wermuth (Wiley-VCH, Weinheim, Germany, 2002).

We have found that the compounds of the present invention or their pharmaceutically acceptable salts are effective inhibitors of BTK and can be used to obtain an inhibitory effect on BTK in a warm-blooded animal in need of such treatment. Accordingly, the compounds of the present invention are expected to be useful in the treatment of diseases or medical conditions mediated in whole or in part by BTK.

Accordingly, the compounds of the present invention are expected to be useful in the treatment of immune disorders, cancer, cardiovascular diseases, viral infections, metabolic/endocrine disorders and neurological disorders, allergic diseases, autoimmune diseases and inflammatory diseases, including urticaria/Sjogren's syndrome, rheumatoid arthritis, osteoporosis, vasculitis, idiopathic thrombocytopenic purpura (ITP), severe myasthenia, allergic rhinitis, asthma, multiple sclerosis and systemic lupus erythematosus.

Based on their BTK inhibitor properties, the compounds of the invention are expected to have a broad spectrum of anticancer properties against BTK-mediated growth observed in human cancer, including but not limited to B-cell neoplasia. In particular, such compounds of the invention are expected to be useful for the treatment of lymphomas and leukemias. More specifically, such compounds of the invention or pharmaceutically acceptable salts thereof are expected to be useful in the treatment of small lymphocytic lymphoma (SLL), follicular lymphoma, Richter's transformation, mantle cell lymphoma, chronic lymphocytic leukemia (CLL), Waldenstrom's macroglobulinemia, non-Hodgkin's lymphoma, primary central nervous system lymphoma, secondary central nervous system lymphoma or diffuse large B-cell lymphoma. In particular, the compounds of the present invention are useful in the treatment of diffuse large B-cell lymphoma with brain metastasis, primary central nervous system lymphoma or secondary central nervous system lymphoma. In particular, the compounds of the present invention are useful in the treatment of chronic lymphocytic leukemia. In particular, the compounds of the present invention are useful in the second-line therapy of chronic lymphocytic leukemia. In particular, the compounds of the present invention are useful in the first-line therapy of chronic lymphocytic leukemia. In particular, the compounds of the present invention are useful in the treatment of diffuse large B-cell lymphoma. In particular, the compounds of the present invention are useful in the treatment of primary central nervous system lymphoma.

In some embodiments, the compounds or pharmaceutically acceptable salts thereof of the present invention have anticancer activity in early stage actively progressing, metastatic and/or pharmacoresistant cancer. In some embodiments, the cancer is locally advanced cancer. In some embodiments, the cancer is locally advanced and/or metastatic cancer. In some embodiments, the cancer is metastatic cancer. In some embodiments, the cancer is invasive cancer. In some embodiments, the cancer is ibrutinib-resistant cancer.

In one embodiment of the invention, where inhibition of BTK is mentioned, this refers to both wild-type BTK and BTK with a C481 mutation (e.g. a C481S, C481Y, C481R or C481F mutation).

In one embodiment of the invention, where inhibition of BTK is mentioned, it refers to wild-type BTK.

In one embodiment of the invention, where inhibition of BTK is mentioned, this refers to BTK with the C481 mutation.

In one embodiment of the invention, where inhibition of BTK is mentioned, this refers to BTK with the C481S mutation.

In one embodiment of the invention, where inhibition of BTK is mentioned, this refers to BTK with the C481Y mutation.

In one embodiment of the invention, where inhibition of BTK is mentioned, this refers to BTK with the C481R mutation.

In one embodiment of the invention, where inhibition of BTK is mentioned, this refers to BTK with the C481F mutation.

Pharmaceutical composition, dosage and administration

The present invention provides pharmaceutical compositions comprising at least one compound of the present invention or a pharmaceutically acceptable salt thereof. In some embodiments, the pharmaceutical composition comprises more than one compound of the present invention or a pharmaceutically acceptable salt thereof. In some embodiments, the pharmaceutical composition comprises one or more compounds of the present invention or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

Typically, pharmaceutically acceptable carriers are conventional carriers for drugs in the art, which can be prepared by methods well known in the pharmaceutical art. In some embodiments, the compounds of the present invention or their pharmaceutically acceptable salts can be mixed with a pharmaceutically acceptable carrier to prepare a pharmaceutical composition.

The form of the pharmaceutical compositions depends on a number of criteria, including but not limited to the route of administration, the degree of the disease, or the dose to be administered. The pharmaceutical compositions can be prepared as preparations for oral, nasal, rectal, transdermal, intravenous, or intramuscular administration. In accordance with the desired route of administration, the pharmaceutical compositions can be prepared as tablets, capsules, pills, dragees, powder, granules, sachets, wafers, lozenges, suspensions, emulsions, solutions, syrups, aerosols (as a solid or in a liquid medium), spray, ointment, paste, cream, lotion, gel, patch, inhaler, or suppositories.

In some embodiments, the pharmaceutical compositions comprise from about 1 mg to about 500 mg of the compounds of the present invention or pharmaceutically acceptable salts thereof, in particular from 1 mg to about 200 mg. The pharmaceutical composition may also be administered one, two, three or even four times a day. However, the daily dose will necessarily vary depending on the host being treated, the particular route of administration and the severity of the disease being treated. Accordingly, the optimal dosage may be determined by the practitioner treating any particular patient.

The therapeutically effective amount of the compound provided herein or its pharmaceutically acceptable salts will depend on various factors known in the art, such as, for example, body weight, age, medical history, medications used, the health condition of the subject and the possibility of cross-reactions, allergic reactions, sensitivities and side effects, as well as the route of administration and the degree of development of the disease. Dosages can be proportionally reduced or increased by a person skilled in the art (e.g., a physician or veterinarian) in accordance with these and other circumstances or requirements.

In a further aspect of the invention there is provided a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof, as defined herein, in combination with a pharmaceutically acceptable diluent or carrier.

In a further aspect of the invention there is provided a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof as defined herein, in combination with a pharmaceutically acceptable diluent or carrier for use in providing an inhibitory effect against BTK in a warm-blooded animal such as a human.

In a further aspect of the invention there is provided a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof as defined herein, in combination with a pharmaceutically acceptable diluent or carrier for use in providing an anti-cancer effect in a warm-blooded animal such as a human.

In a further aspect of the invention there is provided a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof, as defined herein, in combination with a pharmaceutically acceptable diluent or carrier for use in the treatment of small lymphocytic lymphoma (SLL), follicular lymphoma, Richter's transformation, mantle cell lymphoma, chronic lymphocytic leukemia (CLL), Waldenstrom's macroglobulinemia, non-Hodgkin's lymphoma, primary central nervous system lymphoma, secondary central nervous system lymphoma or diffuse large B-cell lymphoma in a warm-blooded animal such as a human.

In a further aspect of the invention there is provided a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof as defined herein, in combination with a pharmaceutically acceptable diluent or carrier for use in the treatment of diffuse large B-cell lymphoma with brain metastases, primary central nervous system lymphoma or secondary central nervous system lymphoma.

Combinations

In some embodiments, the pharmaceutical compositions comprise one or more compounds of the present invention or a pharmaceutically acceptable salt thereof as a first active ingredient and further comprise a second active ingredient. The second active ingredient may be any antitumor agent known in the art, such as PI3K (phosphatidylinositol 3-kinase) inhibitors, anti-CD20 antibodies, anti-PD-1/L1 antibodies, and other approved drugs or combinations of drugs for the treatment of non-Hodgkin's lymphoma. Typical examples of antineoplastic agents as the second active ingredient include, but are not limited to, idelalisib, duvelisib, obinutuzumab, ofatumumab, rituximab, alemtuzumab, bleomycin, brentuximab, vedotin, carmustine, cyclophosphamide, chlorambucil, dacarbazine, dexamethasone, doxorubicin, lomustine, mechlorethamine, procarbazine, prednisone, bendamustine, venetoclax, prednisone, CVP (combination therapy of C - cyclophosphamide, a chemotherapeutic drug, V - vincristine, a chemotherapeutic drug, and P - prednisolone, a steroid), midostaurin, and vinblastine,

It should be understood that the term "combination" as used herein refers to simultaneous, separate or sequential administration. In one aspect of the present invention, "combination" refers to simultaneous administration. In another aspect of the present invention, "combination" refers to separate administration. In a further aspect of the present invention, "combination" refers to sequential administration. If the administration is sequential or separate, the delay in administration of the second component should not be such that the beneficial effect of the combination is lost.

Therefore, in a further aspect the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, as defined herein, in combination with an antitumor agent selected from one of those listed herein above.

Therefore, in a further aspect the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, as defined herein, in combination with an antitumor agent selected from one of those listed herein above, for use in producing an anticancer effect.

Therefore, in a further aspect the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, as defined herein, in combination with an antineoplastic agent selected from one of those listed herein above, for use in the treatment of small lymphocytic lymphoma (SLL), follicular lymphoma, Richter's transformation, mantle cell lymphoma, chronic lymphocytic leukemia (CLL), Waldenstrom's macroglobulinemia, non-Hodgkin's lymphoma, primary central nervous system lymphoma, secondary central nervous system lymphoma or diffuse large B-cell lymphoma.

Therefore, in a further aspect the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, as defined herein, in combination with an antineoplastic agent selected from one of those listed herein above, for use in the treatment of diffuse large B-cell lymphoma with brain metastases, primary central nervous system lymphoma or secondary central nervous system lymphoma.

According to this aspect of the present invention there is provided a combination suitable for use in the treatment of cancer comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof as defined herein and any of the antitumor agents listed above.

According to a further aspect of the present invention there is provided a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof, as defined herein, in combination with an antitumor agent selected from one of those listed herein above, in combination with a pharmaceutically acceptable diluent or carrier.

According to a further aspect of the present invention there is provided a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof, as defined herein, in combination with an antitumor agent selected from one of those listed herein above, in combination with a pharmaceutically acceptable diluent or carrier for use in obtaining an anticancer effect.

According to a further aspect of the present invention there is provided a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof, as defined herein, in combination with an antineoplastic agent selected from one of those listed herein above, in combination with a pharmaceutically acceptable diluent or carrier for use in the treatment of small lymphocytic lymphoma (SLL), follicular lymphoma, Richter's transformation, mantle cell lymphoma, chronic lymphocytic leukemia (CLL), Waldenstrom's macroglobulinemia, non-Hodgkin's lymphoma, primary central nervous system lymphoma, secondary central nervous system lymphoma or diffuse large B-cell lymphoma.

According to a further aspect of the present invention there is provided a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof, as defined herein, in combination with an antineoplastic agent selected from one of those listed herein above, in combination with a pharmaceutically acceptable diluent or carrier for use in the treatment of diffuse large B-cell lymphoma with brain metastases, primary central nervous system lymphoma or secondary central nervous system lymphoma.

According to a further aspect of the present invention, there is provided a kit comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof, as defined herein, in combination with an antitumor agent selected from one of those listed herein above.

According to a further aspect of the present invention, there is provided a kit comprising:

a) a compound of formula (I) or a pharmaceutically acceptable salt thereof, as defined herein, in a first unit dosage form;

b) an antitumor agent selected from one of those listed above, in a second unit dosage form; and

c) a container intended to contain the said first and second dosage forms.

Pharmacological instruments

In addition to their use in therapeutic medicine, the compounds of formula (I) or their pharmaceutically acceptable salts are also useful as pharmacological tools in the development and standardization of in vitro and in vivo test systems for evaluating the effects of inhibition of VTK in laboratory animals such as cats, dogs, rabbits, monkeys, rats and mice, in the search for new therapeutic agents.

Method of treatment

According to a further aspect of the present invention there is provided a compound of formula (I) or a pharmaceutically acceptable salt thereof, as defined herein, for use in a method of treating a human or animal by therapy.

According to a further feature of this aspect of the invention, there is provided a method for providing an inhibitory effect against VTK in a warm-blooded animal, such as a human, comprising administering to said animal an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof, as defined herein.

According to a further feature of this aspect of the invention, there is provided a method of treating cancer in a warm-blooded animal, such as a human, comprising administering to said animal an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof, as defined herein.

According to a further feature of this aspect of the invention, there is provided a method of treating small lymphocytic lymphoma (SLL), follicular lymphoma, Richter's transformation, mantle cell lymphoma, chronic lymphocytic leukemia (CLL), Waldenstrom's macroglobulinemia, non-Hodgkin's lymphoma, primary central nervous system lymphoma, secondary central nervous system lymphoma or diffuse large B-cell lymphoma in a warm-blooded animal such as a human, comprising administering to said animal an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof, as defined herein.

According to a further feature of this aspect of the invention, there is provided a method for treating diffuse large B-cell lymphoma with brain metastases, primary central nervous system lymphoma or secondary central nervous system lymphoma in a warm-blooded animal, such as a human, comprising administering to said animal an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof, as defined herein.

As used herein, the terms "treating" and "treating" refer to reversing, alleviating, delaying the onset, or inhibiting the progression of a disease or disorder, or one or more symptoms thereof, as described herein. In some embodiments, treatment may be administered after one or more symptoms have developed. In other embodiments, treatment may be administered in the absence of symptoms. For example, treatment may be administered to a susceptible individual before symptoms develop (e.g., based on the pattern of symptom development and/or based on genetic or other susceptibility factors). Treatment may also be continued after symptoms have been eliminated, e.g., to prevent or delay their recurrence.

The present invention also provides a method for screening a patient suitable for treatment with a compound of formula (I) or a pharmaceutically acceptable salt thereof, as defined herein. The method comprises sequencing tumor samples of the patients and detecting the accumulation of BTK or the presence of BTK mutations.

According to a further feature of this aspect of the present invention, there is provided a method of treating cancer in a warm-blooded animal, such as a human, comprising (1) determining whether the warm-blooded animal has a cancer susceptible to inhibition of BTK or not and (2) if so, administering to said animal an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof, as defined herein.

Application of connections

In some embodiments, the present invention provides the use of the compounds, their pharmaceutically acceptable salts, or a pharmaceutical composition of the present invention in the manufacture of medicaments for the treatment of diseases or conditions mediated by or dependent on VTK.

Thus, according to this aspect of the invention there is provided a compound of formula (I) or a pharmaceutically acceptable salt thereof, as defined herein, for use as a medicament.

Thus, according to this aspect of the invention there is provided the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof, as defined herein, as a medicament.

Thus, according to this aspect of the invention there is provided a compound of formula (I) or a pharmaceutically acceptable salt thereof, as defined herein, for use in therapy.

According to a further aspect of the invention there is provided the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof as defined herein in the manufacture of a medicament for providing an inhibitory effect against BTK in a warm-blooded animal such as man.

According to this aspect of the invention there is provided the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof as defined herein in the manufacture of a medicament for providing an anti-cancer effect in a warm-blooded animal such as man.

According to a further feature of the invention there is provided the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof as defined herein in the manufacture of a medicament for the treatment of small cell lymphocytic lymphoma (SCLL), follicular lymphoma, Richter's transformation, mantle cell lymphoma, chronic lymphocytic leukemia (CLL), Waldenstrom's macroglobulinemia, non-Hodgkin's lymphoma, primary central nervous system lymphoma, secondary central nervous system lymphoma or diffuse large B-cell lymphoma

According to a further feature of the invention, there is provided the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof, as defined herein, in the manufacture of a medicament for the treatment of diffuse large B-cell lymphoma with brain metastases, primary central nervous system lymphoma or secondary central nervous system lymphoma.

According to a further aspect of the invention there is provided the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof as defined herein for use in providing an inhibitory effect against BTK in a warm-blooded animal such as a human.

According to this aspect of the invention there is provided the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof as defined herein for use in providing an anti-cancer effect in a warm-blooded animal such as a human.

According to a further feature of the invention there is provided a compound of formula (I) or a pharmaceutically acceptable salt thereof as defined herein for use in the treatment of small lymphocytic lymphoma (SLL), follicular lymphoma, Richter's transformation, mantle cell lymphoma, chronic lymphocytic leukemia (CLL), Waldenstrom's macroglobulinemia, non-Hodgkin's lymphoma, primary central nervous system lymphoma, secondary central nervous system lymphoma or diffuse large B-cell lymphoma

According to a further feature of the invention, there is provided a compound of formula (I) or a pharmaceutically acceptable salt thereof, as defined herein, for use in the treatment of diffuse large B-cell lymphoma with brain metastases, primary central nervous system lymphoma or secondary central nervous system lymphoma.

The above features of pharmaceutical compositions, methods, uses and preparation of medicaments also apply to alternative and preferred embodiments of the compounds of the present invention described herein.

EXAMPLES

General experiments

Abbreviations

The synthesis of the compounds provided herein, including pharmaceutically acceptable salts thereof, is illustrated in the synthetic schemes in the examples. The compounds provided herein can be prepared using any known methods of organic synthesis and can be synthesized according to any of numerous possible synthetic routes, and thus these schemes are illustrative only and are not intended to limit other possible methods that can be used to prepare the compounds provided herein. In addition, the steps in the methods are intended for better illustration and can be modified as appropriate. Embodiments of the compounds in the examples were synthesized for research purposes and possibly for submission to regulatory agencies.

The reactions for preparing the compounds of the present invention can be carried out in suitable solvents, which can be readily selected by one skilled in the art of organic synthesis. Suitable solvents can be substantially inert with respect to the starting materials (reactants), intermediates or products at the temperatures at which the reactions are carried out, for example at temperatures that can range from the freezing point of the solvent to the boiling point of the solvent. The reaction can be carried out in a single solvent or in a mixture of more than one solvent. Depending on the particular step of the reaction, the skilled artisan can select suitable solvents for a particular step of the reaction.

The preparation of the compounds of the present invention may involve the protection and deprotection of various chemical groups. The need for protection and deprotection and the selection of appropriate protecting groups can be readily determined by one skilled in the art. The chemistry of protecting groups can be found, for example, in T. W. Greene and P. G. M. Wuts, Protective Groups in Organic Synthesis, 3rd Ed., Wiley & Sons, Inc., New York (1999), which is incorporated herein by reference in its entirety.

The progress of the reactions can be monitored by any suitable method known in the art. For example, product formation can be monitored by spectroscopic methods such as nuclear magnetic resonance spectroscopy (e.g. ¹ H or ¹³ C), infrared spectroscopy, spectrophotometry (e.g. UV and visible), mass spectrometry, or by chromatographic methods such as high performance liquid chromatography (HPLC), liquid chromatography-mass spectroscopy (LC-MS), or thin layer chromatography (TLC). The compounds can be purified by those skilled in the art by a variety of methods, including high performance liquid chromatography (HPLC) ("Preparative LC-MS Purification: Improved Compound Specific Method Optimization") Karl F. Blom, Brian Glass, Richard Sparks, Andrew P. Combs J. Combi. Chem. 2004, 6(6), 874-883, which is incorporated herein by reference in its entirety), supercritical fluid chromatography (SFC) and normal phase chromatography on silica.

The structures of the compounds in the examples were characterized by nuclear magnetic resonance (NMR) and/or liquid chromatography-mass spectrometry (LC-MS). NMR chemical shift (δ) is given in units of 10 ^-6 (ppm). ¹ H-NMR spectra were recorded in dimethyl sulfoxide-d6 (DMSO-d6) solvent, or CDCl ₃ , or CD ₃ OD, or D ₂ O, or acetone-d6, or CD ₃ CN (from Aldrich or Cambridge Isotope Lab., Inc.) on Bruker AVANCE NMR spectrometers (400 MHz) using ICON-NMR (controlled by TopSpin software) or Varian 400MR NMR or Varian VNMR400 NMR spectrometers (400 MHz) (controlled by VnmrJ software) with tetramethylspondyl as an internal standard.

The MS measurement is carried out using a Shimadzu 2010 mass spectrometer, or an Agilent 6110A MSD, or a 1969A time-of-flight mass spectrometer using electrospray, chemical ionization and electron impact ionization methods using a variety of instruments. The detailed methods used in this invention include:

LC-MS method A: 10-80AB_7min_220&254_Shimadzu.lcm

Mobile phase: 1.5 mL/4 L TFA in water (solvent A) and 0.75 mL/4 L TFA in acetonitrile (solvent B), using a gradient elution of 10%-80% (solvent B) over 6 min and holding at 80% for 0.5 min at a flow rate of 0.8 mL/min;

Column: Xtimate C18 2.1×30 mm, 3 µm;

Wavelength: UV 220nm, 254nm;

Column temperature: 50°C;

MS ionization: ESI (electrospray ionization).

LC-MS method B: 10-80AB_4min_220&254_Shimadzu.lcm

Mobile phase: 1.5 mL/4 L TFA in water (solvent A) and 0.75 mL/4 L TFA in acetonitrile (solvent B), using a gradient elution of 10%-80% (solvent B) over 3 min and holding at 80% for 0.5 min at a flow rate of 0.8 mL/min;

Column: Xtimate C18 2.1×30 mm, 3 µm;

Wavelength: UV 220nm, 254nm;

Column temperature: 50°C;

MS ionization: ERI.

LC-MS method C: 10-80CD_7min_220&254_Agilent.lcm

Mobile phase: 0.2 ml/1 l _NH3⋅H2O in water (solvent A) _and acetonitrile (solvent B), using a gradient elution of 10%-80% (solvent B) over 6 min and holding at 80% for 0.5 min at a flow rate of 0.8 ml/min;

Column: Xbrige Shield RP-18, 5 µm, 2.1×50 mm;

Wavelength: UV 220 nm and 254 nm;

Column temperature: 30°C;

MS ionization: ERI.

The measurement by high performance liquid chromatography (HPLC) method is carried out on Shimadzu LC-20A or Shimadzu LC-2010HT series, or Agilent 1200 LC, or Agilent 1100 series systems using Ultimate XB-C18 column (3.0×50 mm, 3 μm or 3.0×150 mm, 3 μm), or Xbridge shieldRP18 column (5 μm, 50 mm×2.1 mm), or Xtimate C18 column (3 μm, 2.1×30 mm), or MERCK RP18 2.5-2 mm, etc. The detailed methods used in the present invention include:

HPLC method A: 10-80AB_8min.met

Mobile phase: 2.75 mL/4 L TFA in water (solvent A) and 2.5 mL/4 L TFA in acetonitrile (solvent B), using a gradient elution of 10%-80% (solvent B) over 6 min and holding at 80% for 2 min at a flow rate of 1.2 mL/min;

Column: Ultimate C18 3.0×50 mm, 3 µm;

Wavelength: UV 220nm, 215nm, 254nm;

Column temperature: 40°C.

HPLC method B: 10-80CD_8min.met

Mobile phase: 2.0 mL/4 L _NH3⋅H2O in water (solvent A) _and acetonitrile (solvent B), using a gradient elution of 10%-80% (solvent B) over 4 min and holding at 80% for 2 min at a flow rate of 1.2 mL/min;

Column: Xbrige Shield RP-18, 2.1×50 mm, 5 µm;

Wavelength: UV 220nm, 215nm, 254nm;

Column temperature: 40°C.

The measurement by supercritical fluid chromatography (SFC) method is carried out on Agilent 1260 series, or Waters UPCC series, or Shimadzu LC-20AB series instruments using ChiralPak AD-3 column (3 μm, 150×4.6 mm), or Chiralcel OJ-3 column (3 μm, 150×4.6 mm), or Chiralpak IG-3 column (3 μm, 50 mm×4.6 mm), etc. The detailed methods used in the present invention include:

SFC method A: mobile phase: A: _CO2 , B: ethanol (0.05% DEA); gradient: 5% to 40% B in 5.5 min and hold at 40% for 3 min, then 5% B for 1.5 min; flow rate: 2.5 ml/min;

Column: ChiralPak AD-3 150×4.6 mm ID. (internal diameter), 3 µm;

Column temperature: 40°C;

Back pressure: 100 bar (10 MPa).

SFC method B: mobile phase: A: _CO2 , B: methanol (0.05% DEA); gradient: 5% to 40% B in 5 min and 40% to 5% B in 0.5 min, hold at 5% B for 1.5 min; flow rate: 2.5 ml/min

Column: Chiralcel OJ-3 150×4.6 mm I.D., 3 µm;

Column temperature: 35°C;

ABPR (Adjustable Back Pressure): 1500 psi (10.34 MPa).

SFC method C: mobile phase: A: _CO2 , B: methanol (0.05% DEA), isocratic elution: 40% B, flow rate: 4 ml/min;

Column: Chiralpak IG-3 50 mm×4.6 mm I.D., 3 µm;

Column temperature: 35°C;

ABPR: 1500 psi (10.34 MPa).

Thin layer chromatography is carried out using Yantai Huanghai HSGF254 silica gel or Anhui Liang Chen Gui Yuan plates. Silica gel plates used for thin layer chromatography (TLC) have a layer thickness of 0.15 mm to about 0.2 mm. Silica gel plates used for separation and purification of products by TLC have a layer thickness of 0.4 mm to about 0.5 mm.

The chromatographic column for purification uses silica gel (100~200, 200~300 or 300~400 mesh, manufactured by Yantai Huanghai Co. or Anhui Liang Chen Gui Yuan Co., etc.) or flash column (40-60 μm silica-CS flash column or 20-35 μm reversed-phase C18 column, manufactured by Agela Technologies, etc.) or silica-CS flash column (40-60 μm) or C18 column (20-40 μm) from Agela Technologies in Teledyne ISCO combination flash system or Biotage flash system as the carrier. The size of the columns is adjusted according to the number of compounds.

The known starting materials of the present invention can be synthesized using or according to methods known in the art, or can be purchased from Alfa Aesar, TCI, Sigma-Aldrich, Bepharm, and Scochem (or PharmaBlock, Bide, Amatek, Stru Chem, Firster Pharmaceutical, Titan (Adamas), etc.).

Unless otherwise stated, all reactions are carried out under an argon or nitrogen atmosphere. An argon or nitrogen atmosphere means that the reaction flask is connected to a cylinder of argon or nitrogen with a volume of about 1 liter. Hydrogenation is usually carried out under pressure. Unless otherwise stated, the reaction temperature in the examples corresponds to the ambient temperature, which is 10 ° C ~ 30 ° C.

The progress of the reactions is monitored by TLC and/or LC-MS. Eluent systems used for the reactions include dichloromethane-methanol and petroleum ether-ethyl acetate. The volume ratios of the solvents are adjusted according to the different polarities of the compounds.

The column chromatography elution system used for compound purification and the TLC eluent system include dichloromethane-methanol system and petroleum ether-ethyl acetate system. The volume ratios of solvents are adjusted according to the different polarity of the compounds. A small amount of alkaline or acidic agents (0.1%~1%) such as formic acid or acetic acid or TFA or ammonia can be added for adjustment.

Compounds of the invention

Methods of synthesis

The compounds of the invention can be prepared according to the following two synthetic methods.

Method A

Compound 2A: Ethyl 5-(((3-chloropyrazin-2-yl)methyl)carbamoyl)tetrahydro-2H-pyran-2-carboxylate

To a mixture of 6-(ethoxycarbonyl)tetrahydro-2H-pyran-3-carboxylic acid (see WO 2019001420A1) (5.90 g, 29.17 mmol) and compound 1A (5.25 g, 29.17 mmol) in dichloromethane (150 mL) were added HATU (16.64 g, 43.76 mmol) and DIEA (11.31 g, 87.51 mmol). The mixture was stirred at room temperature (18-22 °C) for 12 h. The reaction mixture was concentrated and diluted with water (100 mL), extracted with ethyl acetate (200 mL×3). The combined organic layers were washed with brine (200 mL×4), dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was purified by silica gel column chromatography (2.5-3.5% methanol in dichloromethane) to afford compound 2A (9.0 g; yield 94.0%) as a yellow oil.

LC-MS: t _R (retention time) is 0.689 min in the chromatographic method 5-95AB_1.5min_220&254_Shimadzu.lcm (Agilent Pursuit 5 C18 20×2.0 mm), MS (ESI) m/z is 328.2 [M+H] ⁺ .

Compound 3A: ethyl 5-(8-chloroimidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-carboxylate

To a mixture of compound 2A (9.0 g, 27.46 mmol) and DMF (600 µL) in MeCN (150 mL) was added _POCl3 (21.05 g, 137.30 mmol). The mixture was stirred at 70 °C for 1 h. The reaction mixture was concentrated and washed with water (50 mL) and saturated _NaHCO3 (50 mL), extracted with ethyl acetate (100 mL×3). The combined organic layers were washed with brine (200 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was purified by silica gel column chromatography (52~62% ethyl acetate in petroleum ether) to give compound 3A (3.5 g, 41.2% yield) as a yellow oil.

LC-MS: t _R is 0.742 min in the chromatographic method 5-95AB_1.5min_220&254_Shimadzu.lcm (Agilent Pursuit 5 C18 20×2.0 mm), MS (ESI) m/z is 310.2 [M+H] ⁺ .

¹ H NMR (400 MHz, CDCl ₃ ): δ 7.80 (d, J=0.4 Hz, 1H), 7.70-7.64 (m, 1H), 7.37 (d, J=4.8 Hz, 0.3H), 7.35 (d, J=4.8 Hz, 0.7H), 4.44 (t, J=4.4 Hz, 0.7H), 4.30-4.25 (m, 2H), 4.23-4.14 (m, 1H), 4.00 (dd, J=3.2, 12.0 Hz, 1H), 3.79 (t, J=11, 2 Hz, 0.4H), 3.42-3.19 (m, 1H), 2.47-2.37 (m, 0.8N), 2.29-2.06 (m, ZN), 1.91-1.77 (m, 0.4N), 1.35-1.31 (m, 3N).

Compound 4A: (5-(8-chloroimidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol

To a mixture of LiAlH ₄ (860 mg, 22.60 mmol) in THF (20 mL) was added compound 3A (3.5 g, 11.30 mmol) in THF (20 mL) at 0 °C. The mixture was stirred at 0 °C for 1 h. The reaction mixture was quenched with water (860 μL), 15% NaOH (860 μL), and water (2580 μL). The mixture was dried over anhydrous sodium sulfate and stirred at room temperature for 0.5 h, filtered, and concentrated to give compound 4A (2.7 g, 89.4% yield) as a yellow solid.

LC-MS: t _R is 0.635 min in the chromatographic method 5-95AB_1.5min_220&254_Shimadzu.lcm (Agilent Pursuit 5 C18 20×2.0 mm), MS (ESI) m/z is 267.8 [M+H] ⁺ .

Compound 5A: (5-(1-bromo-8-chloroimidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol

To a mixture of compound 4A (2.7 g, 10.11 mmol) in MeCN (100 mL) was added NBS (2.34 g, 13.14 mmol). The mixture was stirred at room temperature (16-20 °C) for 1 h. The reaction mixture was concentrated to give the crude product, which was purified by silica gel column chromatography (2.2% methanol in dichloromethane) to give compound 5A (2.5 g, 71.63% yield) as a yellow solid.

LC-MS: t _R is 0.717 min in the chromatographic method 5-95AB_1.5min_220&254_Shimadzu.lcm (Agilent Pursuit 5 C18 20×2.0 mm), MS (ESI) m/z is 347.8 [M+H] ⁺ .

Compound 6A: (5-(8-amino-1-bromimidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol

To a mixture of compound 5A (2.5 g; _7.21 mmol) in IPA (15 mL) was added _NH3⋅H2O (15 mL). The mixture was stirred at 100°C for 12 h in a sealed 100 mL tube. The reaction mixture was concentrated to give compound 6A (2.4 g; 98.0% purity) as a yellow oil.

LC-MS: t _R is 0.436 min in the chromatographic method 10-80AB_3min_220&254_Shimadzu.lcm (Xtimate C18 2.1×30 mm), MS (ESI) m/z is 327.1 [M+H] ⁺ .

Compound 7A: (5-(8-amino-1-(2-fluoro-4-phenoxyphenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol

To a mixture of compound 6A (500 mg, 1.31 mmol, purity 97.99%) and (2-fluoro-4- _{phenoxyphenyl} )boronic acid (450 mg, 1.95 mmol) in 1,4-dioxane (15 mL)/ _H2O (5 mL) was added Pd(dppf) _Cl2⋅CH2Cl2 (32 mg, 0.04 mmol) and _K2CO3 (375 mg, 2.62 mmol) under nitrogen atmosphere. The mixture was stirred at 100 °C for 12 h under nitrogen atmosphere. The reaction _mixture was diluted with water (20 mL), extracted with ethyl acetate (50 mL×3). The combined organic layers were washed with brine ( ₁₀₀ mL), dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. The crude product was purified by silica gel column chromatography (2.2% methanol in dichloromethane) to give compound 7A (cis/trans racemate mixture) (150 mg; yield 26.4%) as a yellow solid.

LC-MS: t _R is 0.723 min in the chromatographic method 5-95AB_1.5min_220&254_Shimadzu.lcm (Agilent Pursuit 5 C18 20×2.0 mm), MS (ESI) m/z is 435.1 [M+H] ⁺ .

(2-fluoro-4-phenoxyphenyl)6oronic acid:

To a mixture of compound 1a (10 g, 52.35 mmol) in _CH2Cl2 (480 mL ₎ was added PhB(OH)2 (12.8 g, 104.71 mmol), Cu(OAc) ₂ (9.5 g, 52.35 mmol), _NEt3 (21 mL, 151.05 mmol) and 4Å MS (molecular sieve) (5 g) at room temperature (25-30 °C). The mixture was stirred at room temperature (25-30 °C) for 16 h under air atmosphere. The mixture was filtered through a pad of celite. The filtrate was concentrated in vacuo to give the crude material, which was purified by column chromatography on silica gel (petroleum ether) to give compound 2a (11.3 g, yield 80.8%) as a colorless oil.

¹ H NMR (400 MHz, CDCl ₃ ): δ 7.47 (t, J=8.4 Hz, 1H), 7.42-7.36 (m, 2H), 7.19 (t, J=8.4 Hz, 1H), 7.05 (d, J=8.0 Hz, 2H), 6.78 (dd, J=10.0, 2.8 Hz, 1H), 6.71 (dd, J=8.8, 2.0 Hz, 1H).

To a solution of compound 2a (11.3 g, 42.30 mmol) in THF (150 mL) at -65 °C was added n-BuLi (19 mL, 46.53 mmol, 2.5 N in n-hexane) at -65 °C. The mixture was stirred at -65 °C for 0.5 h. Then B(Oi-Pr) ₃ (9.5 g, 50.76 mmol) was added at -65 °C. The mixture was stirred at -65 °C for 2 h. The mixture was quenched with saturated ammonium chloride solution (50 mL), extracted with ethyl acetate (50 mL×3), washed with brine (100 mL×2). The combined organic layer was dried over anhydrous sodium sulfate, filtered and concentrated to give a crude material which was triturated with petroleum ether (100 mL), filtered and the filter cake was concentrated to give (2-fluoro-4-phenoxyphenyl)boronic acid (4.3 g) as a yellow oil. The filtrate was concentrated and purified by silica gel column chromatography (0~20% ethyl acetate in petroleum ether) to give 2-fluoro-4-phenoxyphenyl)boronic acid (3 g) as a yellow oil.

¹ H NMR (400 MHz, DMSO-d6): δ 8.10 (s, 2H), 7.58 (t, J=8.0 Hz, 1H), 7.47-7.41 (m, 2H), 7.22 (t, J=7.6 Hz, 1H), 7.09 (d, J=7.6 Hz, 2H), 6.78-6.68 (m, 2H).

Trans-isomer 1, compound 2: trans-(5-(8-amino-1-(2-fluoro-4-phenoxyphenyl)-imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol

Trans-isomer 2, compound 3: trans-(5-(8-amino-1-(2-fluoro-4-phenoxyphenyl)-imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol

A mixture of compound 7A (150 mg; 0.35 mmol) _{was purified by chiral SFC (column: DAICEL CHIRALPAK AD-H) (250 mm×30 mm, 10 μm); conditions: 0.1% NH3⋅H2O in} EtOH, 40%, flow rate 80 mL/min). Fractions containing the desired compounds were concentrated, diluted with _H2O (10 mL) and _CH3CN (10 mL), lyophilized to afford compound 2 (34.8 mg; yield 23.2%) as a white solid and compound 3 (29.4 mg; 19.6%) as a white solid. The two cis-enantiomers were not collected.

Spectra of compound 2:

LC-MS: t _R is 2.037 min in the chromatographic method 10-80AB_7min_220&254_Shimadzu.lcm (Xtimate C18 2.1×30 mm), MS (ESI) m/z is 435.3 [M+H] ⁺ .

HPLC: t _R is 3.03 min in 10-80AB_8min.met (HPLC-BJ Ultimate 3.0×50 mm 3μm).

SFC: t _R is 6.119 min, optical purity: 96.3%. Method details: Column: ChiralPak AD-3 150×4.6 mm ID, 3 μm; Mobile phase: A: CO ₂ , B: ethanol (0.05% DEA); Gradient: 5% to 40% B over 5.5 min and hold at 40% for 3 min, then 5% B for 1.5 min; Flow rate: 2.5 mL/min; Column temperature: 40°C; Back pressure: 100 bar (10 MPa).

¹ H NMR (400 MHz, CD ₃ OD) δ 7.63 (d, J=5.2 Hz, 1H), 7.54-7.36 (m, 3H), 7.27-7.18 (m, 1H), 7.14 (dd, J= 8.8, 0.8 Hz, 2H), 7.02 (d, J=4.8 Hz, 1H), 6.98-6.82 (m, 2H), 4.23-4.07 (m, 1H), 3.75 (t, J=11.2 Hz, 1H), 3.62-3.51 (m, 3H), 3.46-3.36 ( m, 1H), 2.25-2.15 (m, 1H), 2.13-1.96 (m, 1H), 1.79 (d, J=13.6 Hz, 1H), 1.66-1.49 (m, 1H).

¹⁹ F HMP (CD ₃ OD) δ -112.240.

Spectra of compound 3:

LC-MS: t _R is 2.030 min in the chromatographic method 10-80AB_7min_220&254_ Shimadzu.lcm (Xtimate C18 2.1×30 mm), MS (ESI) m/z is 435.3 [M+H] ⁺ .

HPLC: t _R is 3.02 min in 10-80AB_8min.met (HPLC-BJ Ultimate 3.0×50 mm 3 µm).

SFC: t _R is 7.334, optical purity: 92.7%. Method details: Column: ChiralPak AD-3 150×4.6 mm ID, 3 μm; Mobile phase: A: _CO2 , B: ethanol (0.05% DEA); Gradient: 5% to 40% B over 5.5 min and hold at 40% for 3 min, then 5% B for 1.5 min; Flow rate: 2.5 mL/min; Column temperature: 40°C; Back pressure: 100 bar (10 MPa).

¹ H NMR (400 MHz, CD ₃ OD) δ 7.65 (d, J=5.2 Hz, 1H), 7.52-7.37 (m, 3H), 7.29-7.21 (m, 1H), 7.20-7.11 (m, 2H), 7.04 (d,.7=4.8 Hz, 1H), 6.99-6.84 (m, 2H), 4.24-4.13 (m, 1H), 3.77 (t, J=11.2 Hz, 1H), 3.62-3.51 (m, 3H), 3.48-3.77 (m, 1H), 2.31-2.17 (m, 1H), 2.15-2.01 (m, 1H), 1.91-1.74 (m, 1H), 1.69-1.51 (m, 1H).

¹⁹ F HMP (400 MHz, CD ₃ OD) δ -112.258.

The following compounds were synthesized according to method A:

METHOD B

Compound 2B: Ethyl 5-(((3-amino-5-hydroxy-1,2,4-triazin-6-yl)methyl)carbamoyl)-tetrahydro-2H-pyran-2-carboxylate

To a mixture of 5-(2,5-dioxopyrrolidin-1-yl)-2-ethyl tetrahydro-2H-pyran-2,5-dicarboxylate (20 g, crude; 60.88 mmol) in acetonitrile (30 mL) was added compound 1B (13 g, 60.88 mmol) and triethylamine (25 mL, 182.6 mmol). The mixture was stirred at 50 °C for 16 h. The mixture was concentrated to give compound 2B (35 g, crude) as a brown solid.

LC-MS: t _R is 0.582 min in the chromatographic method 5-95AB_1.5min_220&254_Shimadzu.lcm (Merck RP18 25-3 mm), MS (ESI) m/z is 325.9 [M+H] ⁺ .

5-(2,5-dioxopyrrolidin-1-yl)-2-ethyl tetrahydro-2H-pyrap-2,5-dicarboxylate:

To a solution of 6-(ethoxycarbonyl)tetrahydro-2H-pyran-3-carboxylic acid (1.4 g, 6.92 mmol) in _CH2Cl2 (30 _mL ) were added 1-hydroxypyrrolidin-2,5-dione (877 mg, 7.61 mmol) and EDCI (1.6 g, 8.30 mmol). The mixture was stirred at 22-26 °C for 1.5 h. The mixture was diluted with dichloromethane (30 mL), washed with brine (50 mL×3). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated to give 5-(2,5-dioxopyrrolidin-1-yl)-2-ethyl tetrahydro-2H-pyran-2,5-dicarboxylate as a colorless oil, which was used directly.

Compound 3B: Ethyl 5-(2-amino-4-hydroxyimidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-carboxylate

To a solution of compound 2B (35 g, crude; 60.88 mmol) in acetonitrile (300 mL) was added phosphoric acid chloride (23 mL, 243.52 mmol). The mixture was then stirred at 70 °C for 16 h. The mixture was concentrated, the residue was diluted with dichloromethane (150 mL), poured into cooled saturated sodium bicarbonate solution (300 mL) until pH 8, extracted with dichloromethane/methanol (10:1; 200 mL×4) followed by extraction with IPA/ _CHCl3 (150 mL×4). The combined organic layer was dried over anhydrous sodium sulfate, filtered and concentrated to give compound 3B (45 g, crude) as a black oil.

LC-MS: t _R is 0.13 min in the chromatographic method 5-95AB_1.5min_220&254_Shimadzu.lcm (Merck RP18 25-3 mm), MS (ESI) m/z is 308.0 [M+H] ⁺ .

Compound 4B: Ethyl 5-(2-amino-5-bromo-4-hydroxyimidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-carboxylate

To a solution of compound 3B (45 g, crude; 60.88 mmol) in N,N-dimethylformamide (200 mL) was added NBS (11.8 g, 66.96 mmol). The mixture was stirred at room temperature (16-21 °C) for 0.5 h. The mixture was poured into water (300 mL), extracted with ethyl acetate (300 mL×4), washed with brine (500 mL×4). The combined organic layer was dried over anhydrous sodium sulfate, filtered and concentrated to give compound 4B (19 g, crude) as a black oil.

LC-MS: t _R is 0.602 min in the chromatographic method 5-95AB_lmin_220&254_Agilent chromatography (Agilent Poroshell 120 EC-C18 2.7 μm 3.0×30 mm), MS (ESI) m/z is 388.0 [M+H+2] ⁺ (Bromo isotope).

Compound 5B: Ethyl 5-(5-6ro-4-hydroxyimidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-carboxylate

To a solution of compound 4B (19 g, crude; 49.19 mmol) in tetrahydrofuran (300 mL) was added tert-butyl nitrite (12 mL; 98.39 mmol) at 0-5 °C, and the mixture was stirred at room temperature (16-21 °C) for 16 h. The mixture was combined with another portion (7.2 g of crude compound 4B) and concentrated to give a crude material, which was purified by silica gel column chromatography (0-60% ethyl acetate in petroleum ether) to give compound 5B (14.3 g; 44% yield over 5 steps) as a yellow solid.

LC-MS: t _R is 0.753 min in the chromatographic method 5-95AB_1.5min_220&254_Shimadzu.lcm (Agilent Pursuit 5 C18 20x×2.0 mm), MS (ESI) m/z is 370.9 [M+H] ⁺ .

¹ H NMR (400 MHz, CD ₃ OD): δ 7.74 (s, 0.3H), 7.72 (s, 0.5H), 4.37 (t, J=4.8 Hz, 0.7H), 4.29-4.16 (m, 3H), 4.12 (dd, J=11.6, 2.0 Hz, 0.3H), 3.92 (dd, J=11.6, 4.0 Hz, 0.7H), 3.70 (t, J=11.2 Hz, 0.4H), 3.57-3.45 (m, 1H), 2.35-1.92 (m, 4H), 1.33-1.27 (m, 3H).

Compound 6B: Ethyl 5-(4-amino-5-bromimidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-carboxylate

To a solution of 1,2,4-triazole (28 g; 404.1 mmol) in acetonitrile (200 ml) was added POCl ₃ (12.5 ml; 134.7 mol) at a temperature below 10 °C followed by the addition of trimethylamine (56 ml; 404.1 mmol). The mixture was stirred for 20 min at 10 °C, compound 5B (5 g; 13.47 mmol) was added and the reaction mixture was stirred for 1.5 h at 90 °C. The mixture was cooled to 10 °C, ammonia (30 ml; 28%) was added maintaining the temperature below 20 °C and stirred for 0.5 h at 10 °C. Another batch was prepared on the same scale. The mixtures were combined and diluted with water (200 mL) and extracted with ethyl acetate (500 mL×3), washed with brine (500 mL×3), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo, then purified by silica gel column chromatography (2-4% methanol in dichloromethane) to give compound 6B (9 g; 90% yield) as a yellow solid.

LC-MS: t _R is 0.614 min in the chromatographic method \5-95AB_lmin_220&254_Agilent (Agilent Poroshell 120 EC-C18 2.7 μm 3.0×30 mm), MS (ESI) m/z is 372.0 [M+H+2] ⁺ (Bromo isotope).

¹ H NMR (400 MHz, CD ₃ OD) _: δ 7.81 (s, 0.3H), 7.79 (s, 0.6H), 4.36 (t, J=5.2 Hz, 0.6H), 4.29-4.19 (m, 3H), 4.12 (dd, J=11.6, 2.0 Hz, 0.3H), 3.93 (dd, J=11.6, 4.0 Hz, 0.7H), 3.71 (t, J=11.2 Hz, 0.3H), 3.63-3.48 (m, 1H), 2.35-2.20 (m, 1H), 2.17-1.94 (m, 2.7H), 1.79-1.68 (m, 0.3H), 1.34-1.26 (m, 3H).

Compound 7B: (5-(4-amino-5-bromimidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-yl)methanol

To a solution of compound 6B (6.1 g, 16.48 mmol) in tetrahydrofuran (120 mL) cooled at 0-5 °C was added LiAlH ₄ (1.20 g, 32.95 mmol), maintaining the temperature below 10 °C. The mixture was stirred at 0-10 °C for 1.5 h. To the mixture was added 25 g Na ₂ SO ₄ ⋅10H ₂ O at 0-5 °C, stirred for 2 h, and filtered. The filter cake was suspended twice in dichloromethane/methanol (100 mL of a 10:1 mixture) and filtered. The combined filtrate was concentrated in vacuo and the residue was purified by silica column chromatography (0~10% methanol in dichloromethane) to afford compound 7B (3.9 g; 72% yield) as a white solid and 0.6 g of the debromination by-product as a yellow solid.

LC-MS: t _R is 1.958 and 2.016 min in the chromatographic method \0-60AB_7min_220&254_Shimadzu.lcm (Xtimate C18 2.1×30 mm, 3 μm), MS (ESI) m/z is 328.1 [M+H] ⁺ .

¹ H NMR (400 MHz, methanol-d ₄ ) _: δ 7.80 (s, 0.3H), 7.78 (s, 0.6H), 4.47 (dt, J=10.0, 2.0 Hz, 0.6H), 4.20-4.13 (m, 0.3H), 3.84 (dd, J=11.6, 3.2 Hz, 0.6H), 3.64 (t, J=10.8 Hz, 0.4H), 3.60-3.38 (m, 4H), 2.45-2.36 (m, 0.6H), 2.22-2.13 (m, 0.3H), 2.09-1.96 (m, 1H), 1.93-1.73 (m, 1H), 1.62-1.45 (m, 1H).

Compound 8B: (5-(4-amino-5-(2-fluoro-4-phenoxyphenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-yl)methanol

Compound 7B (106 mg; 0.457 mmol), Pd( _dppf ) _Cl2⋅CH2Cl2 (25 mg; 0.0304 mmol) and potassium carbonate (84 _mg ; 0.608 mmol) were placed in a reaction tube and purged with nitrogen 3 times, and (2-fluoro-4-phenoxyphenyl)boronic acid (100 mg; 0.304 mmol) in 1,4-dioxane (3 mL) and water (1 mL) were added. The resulting mixture was stirred at 100°C under nitrogen for 2 h. The mixture was concentrated to give a crude material, which was purified by silica gel column chromatography (0-100% ethyl acetate in petroleum ether) to give compound 8B (80 mg, crude; mixture of trans/cis racemates) as a yellow oil.

LC-MS: t _R is 2.274 min and 2.340 min in the chromatographic method 10-80AB_7min_220&254_Shimadzu.lcm (Xtimate C18 2.1×30 mm), MS (ESI) m/z is 436.2 [M+H] ⁺ .

SFC: t _R is 4.171 min, 4.286 min, 4.454 min and 5.581 min. Method: Column: Chiralcel OJ-3 150×4.6 mm ID, 3 μm; Mobile phase: A: CO ₂ , B: methanol; Gradient: 5% to 40% B in 5 min and 40% to 5% B in 0.5 min, hold at 5% B for 1.5 min; Flow rate: 2.5 mL/min, Column temperature: 35°C; ABPR: 1500 psi (10.34 MPa).

Cis-isomer 1, compound 8: cis-(5-(4-amino-5-(2-fluoro-4-phenoxyphenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-yl)methanol

Cis-isomer 2, compound 9: cis-(5-(4-amino-5-(2-fluoro-4-phenoxyphenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-yl)methanol

Trans isomer 1, compound 10: trans-(5-(4-amino-5-(2-fluoro-4-phenoxyphenyl)-imidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-yl)methanol

Trans isomer 2, compound 11: trans-(5-(4-amino-5-(2-fluoro-4-phenoxyphenyl)-imidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-yl)methanol

Compound 8B (80 mg) was further separated by preparative SFC (column: DAICEL CHIRALCEL OJ-H (250 mm×30 mm, 5 μm); conditions: 30% MeOH (0.1% NH3⋅H2O) in _CO2 ; flow rate: 60 mL/min) to give 6.8 mg of crude compound 11, which was further purified by preparative HPLC (column: Welch Xtimate C18 100×40 mm, 3 μm, conditions: 25-55% (A: water (0.225% EA), B: _CH3CN ), flow rate: 25 mL/min) to give compound 11 (1.7 mg; yield 1% over 2 steps) as a white solid. After SFC, the mixture of the other three peaks (40 mg) was further purified by chiral SFC (column: DAICEL CHIRALPAK IG (250 mm×30 mm, 10 μm); conditions: 55% MeOH ( _{0.1% NH3⋅H2O )} in _CO2 ; flow rate: 80 mL/min) to afford compound 8 (8.6 mg; yield 6.4% over 2 steps) as a white solid, compound 9 (6.7 mg; yield 5% over 2 steps) as a white solid, and compound 10 (5.4 mg; yield 4% over 2 steps) as a white solid.

Spectra of compound 8:

LC-MS: t _R is 2.356 min in the chromatographic method 10-80AB_7min_220&254_Shimadzu.lcm (Xtimate C18 2.1×30 mm), MS (ESI) m/z is 436.2 [M+H] ⁺ .

HPLC: t _R is 3.00 min in the chromatographic method 10-80CD_8min.met. (XBridge Shield RP 18 2.1×50 mm, 5 µm).

¹ H NMR (400 MHz, CD ₃ OD): δ 7.84 (s, m), 7.54 (t, J=8.4 Hz, 1H), 7.47-7.41 (m, 2H), 7.26-7.20 (m, 1H ), 7.17-7.12 (m, 2H), 6.94 (dd, J=8.0, 2.0 Hz, 1H), 6.88 (dd, J=10.8, 2.4 Hz, 1H), 4.56-4.51 (m, 1H), 3.89 (dd, J=11.6 , 3.6 Hz, 1H), 3.65-3.48 (m, 4H), 2.51-2.42 (m, 1H), 2.12-2.02 (m, 1H), 1.99-1.85 (m, 1H), 1.64-1.55 (m, 1H).

SFC: t _R is 4.059 min, optical purity is 99.94%.

Method: Column: Chiralcel OJ-3 150×4.6 mm ID, 3 μm; Mobile phase: A: _CO2 , B: methanol (0.05% DEA); Gradient: 5% to 40% B in 5 min and 40% to 5% B in 0.5 min, hold at 5% B for 1.5 min; Flow rate: 2.5 mL/min; Column temperature: 35°C; ABPR: 1500 psi (10.34 MPa).

Spectra of compound 9:

LC-MS: t _R is 2.340 min in the chromatographic method 10-80AB_7min_220&254_Shimadzu.lcm (Xtimate C18 2.1×30 mm), MS (ESI) m/z is 436.3 [M+H] ⁺ .

HPLC: t _R is 2.99 min in the chromatographic method 10-80CD_8min.met. (XBridge Shield RP 18 2.1×50 mm; 5 µm).

¹ H NMR (400 MHz, CD ₃ OD): δ 7.84 (s, m), 7.53 (t, J=8.4 Hz, 1H), 7.47-7.41 (m, 2H), 7.25-7.19 (m, 1H ), 7.17-7.12 (m, 2H), 6.94 (dd, J=8.4, 2.0 Hz, 1H), 6.87 (dd, J=11.2, 2.4 Hz, 1H), 4.56-4.50 (m, 1H), 3.88 (dd, J=12.0 , 3.6 Hz, 1H), 3.65-3.48 (m, 4H), 2.50-2.41 (m, 1H), 2.12-2.02 (m, 1H), 1.97-1.86 (m, 1H), 1.64-1.55 (m, 1H).

SFC: t _R is 4.161 min, optical purity is 100%.

Method: Column: Chiralcel OJ-3 150×4.6 mm ID, 3 μm; Mobile phase: A: _CO2 , B: methanol (0.05% DEA); Gradient: 5% to 40% B in 5 min and 40% to 5% B in 0.5 min, hold at 5% B for 1.5 min; Flow rate: 2.5 mL/min; Column temperature: 35°C; ABPR: 1500 psi (10.34 MPa).

Spectra of compound 10:

LC-MS: t _R is 2.410 min in the chromatographic method 10-80AB_7min_220&254_Shimadzu.lcm (Xtimate C18 2.1×30 mm), MS (ESI) m/z is 436.3 [M+H] ⁺ .

HPLC: t _R is 2.98 min in the chromatographic method 10-80CD_8min.met. (XBridge Shield RP 18 2.1×50 mm, 5 µm).

^1H NMR (400 MHz, CD ₃ OD): δ 7.86 (s, 1H), 7.50 (t, J=8.4 Hz, 1H), 7.47-7.41 (m, 2H), 7.26-7.20 (m, 1H ), 7.17-7.12 (m, 2H), 6.94 (dd, J=8.4, 2.4 Hz, 1H), 6.88 (dd, J=10.8, 2.4 Hz, 1H), 4.24-4.18 (m, 1H), 3.75 (t, J=11.2 Hz, 1H), 3.68-3.48 (m, 4H), 2.27-2.19 (m, 1H), 2.16-2.04 (m, 1H), 1.84-1.76 (m, 1H), 1.59-1.47 (m, 1H).

SFC: t _R is 4.405 min, optical purity is 99.83%.

Method: Column: Chiralcel OJ-3 150×4.6 mm ID, 3 μm; Mobile phase: A: _CO2 , B: methanol (0.05% DEA); Gradient: 5% to 40% B in 5 min and 40% to 5% B in 0.5 min, hold at 5% B for 1.5 min; Flow rate: 2.5 mL/min; Column temperature: 35°C; ABPR: 1500 psi (10.34 MPa).

Spectra of compound 11:

LC-MS: t _R is 2.365 min in the chromatographic method 10-80AB_7min_220&254_Shimadzu.lcm (Xtimate C18 2.1×30 mm), MS (ESI) m/z is 436.2 [M+H] ⁺ .

HPLC: t _R is 3.05 min in the chromatographic method 10-80CD_8min.met. (XBridge Shield RP 18 2.1×50 mm, 5 µm).

^1H NMR (400 MHz, CD ₃ OD): δ 7.86 (s, 1H), 7.50 (t, J=8.4 Hz, 1H), 7.47-7.41 (m, 2H), 7.26-7.21 (m, 1H ), 7.17-7.12 (m, 2H), 6.94 (dd, J=8.4, 2.4 Hz, 1H), 6.88 (dd, J=10.8, 2.4 Hz, 1H), 4.24-4.18 (m, 1H), 3.75 (t, J=10.8 Hz, 1H), 3.68-3.47 (m, 4H), 2.26-2.18 (m, 1H), 2.16-2.04 (m, 1H), 1.84-1.76 (m, 1H), 1.59-1.47 (m, 1H).

SFC: t _R is 5.802 min, optical purity is 100%.

Method: Column: Chiralcel OJ-3 150×4.6 mm ID, 3 μm; Mobile phase: A: _CO2 , B: methanol (0.05% DEA); Gradient: 5% to 40% B over 5 min and 40% to 5% B over 0.5 min, hold at 5% B for 1.5 min; Flow rate: 2.5 mL/min; Column temperature: 35°C; ABPR: 1500 psi (10.34 MPa). The following compounds were synthesized according to Method B:

Biological data

WT BTK and C481S HTRF BTK kinase assay

Recombinant wild-type BTK (WT BTK) was purchased from Thermo fisher. Recombinant BTK(C481S) was purchased from SignalChem. The inhibitory activity of the compounds against BTK and BTK(C481S) was assessed using a time-resolved homogeneous fluorescence assay.

Briefly, recombinant kinases were pre-incubated in the presence or absence of compound at room temperature for 30 min. The reaction was initiated by adding ATP and a peptide substrate that could be phosphorylated by the kinases during the reaction. After incubation for 120 min, the reaction was stopped by adding a detection reagent mixture containing EDTA. Fluorescence was measured at 615 nm and 665 nm, respectively, with an excitation wavelength of 320 nm. The calculated ratio of 665 nm/615 nm signals is proportional to the kinase activity. The concentration of compound providing 50% inhibition of the corresponding kinase (IC ₅₀ ) was calculated using a four-parameter logistic model fit with XL fitting.

ELISA (enzyme-linked immunosorbent assay) of r-BTK cell line TMD8

TMD-8 cells were seeded at a density of 30,000 cells/well in 96-well plates with RPMI1640 medium supplemented with 1.5% fetal bovine serum. Test compounds were added to the cells and the cells were incubated for 0.5 h at 37°C, 5% CO ₂ . Pervanadate solution was then added to the cells to a final concentration of 100 μM and the cells were incubated for another 1 h at 37°C, 5% CO ₂ . Following compound treatment, cells were lysed and the p-BTK signal was detected using a protocol specifically designed for the PathScan® Phospho-Btk Sandwich ELISA (Tyr223) (#23843). The plate was read on a Multiscan Spectrum reader set to 450 nm. These data were processed in GraphPad Prism.

HEK293-BTK(WT) and HEK293-BTK(C481S)

Full-length BTK cDNAs containing the C481S mutation were generated by mutagenesis using the QuikChange II XL Site-Directed Mutagenesis Kit. The mutant BTK cDNAs were confirmed by sequencing. Then, BTK(WT), BTK-C481S cDNAs were cloned into the PLVX-Puro lentiviral vector. Lentiviruses were packaged into 293T cells by transfection with lentiviral vectors and a packaging mixture. BTK(WT), BTK-C481S lentiviruses were transfected into HEK293 cells. Transfected cells were selected in 2 μg/ml puromycin. Stable polyclonal cell lines were confirmed by WB and used for further study. HEK293-BTK(WT), HEK293-BTK(C481S) cells were cultured in DMEM (Gibco; 12430) with 10% FBS (fetal bovine serum) (Gibco; 10099), with 1 μg/ml puromycin. All cells were maintained in a humidity-controlled incubator at 37°C with 5% CO ₂ .

HEK293-BTK(WT) and HEK293-BTK(C481S) cells were seeded at a density of 5,000 cells/well in 96-well plates with RPMI1640 medium supplemented with 1.5% fetal bovine serum. Test compounds were added to the cells and the cells were incubated for 1.5 h at 37°C, 5% CO ₂ . Following compound treatment, cells were lysed and the p-BTK signal was detected according to the exact assay of the PathScan® Phospho-Btk (Tyr223) Sandwich ELISA Kit (#23843). The plate was read on a Multiscan Spectrum reader set to 450 nm. Data were analyzed using GraphPad Prism software.

TMD-8 Anti-Proliferation Assay

TMD-8 cells were prepared in RPMI1640 medium containing 10% fetal bovine serum and seeded in 384-well plates at a density of 1500 cells per well. Cells were incubated overnight at 37°C, 5% CO ₂ . After incubation, compounds were added to the assay plates at different concentrations and the cells were incubated for another 72 hours at 37°C, 5% CO ₂ . After incubation for 72 hours, 15 μl of CellTiter 96® AQueous One Solution Reagent was added to each well and the plates were incubated at room temperature for 30 minutes. Cell viability was determined using the CellTiter-Glo kit (Promega, USA). The CellTiter-Glo assay was performed according to the manufacturer's instructions and luminescence was determined in a multimode plate reader (Envision, PerkinElmer, USA). Data were processed in XLfit software.

In vitro rat/human hepatocyte clearance assay

Male rat hepatocytes and human hepatocytes of both sexes were obtained from commercial suppliers (e.g., BioreclamationlVT) and stored at -150°C until use. 10 mM stock solutions of test compounds were prepared in DMSO. Thawing medium and incubation medium with supplements (serum-free) were placed in a water bath at 37°C for at least 15 min before use. Stock solutions were diluted to 100 μM by combining 198 μL of acetonitrile and 2 μL of 10 mM stock solution.

Vials containing cryopreserved hepatocytes were removed from storage, ensuring that the vials remained at cryogenic temperatures. The vials were thawed in a 37°C water bath with gentle shaking. The vials were kept in the water bath until all ice crystals had dissolved and disappeared. The vials were sprayed with 70% ethanol before being transferred to a biological safety cabinet. The contents were then poured into a 50-mL conical tube containing thawing medium. The vials were centrifuged at 100 g for 10 min at room temperature. The thawing medium was aspirated, and the hepatocytes were resuspended in serum-free incubation medium to yield approximately 1.5 × 10 ⁶ cells/mL.

Cell viability and density were quantified using trypan blue exclusion and cells were then diluted with serum-free incubation medium to a working cell density of 1 x 10 ⁶ viable cells/mL. A subset of hepatocytes at 1 x 10 ⁶ viable cells/mL were boiled for 10 min before being added to the plate as a negative control to eliminate enzymatic activity so that little or no substrate conversion was observed. Inactivated hepatocytes were used to prepare negative samples, which were used to eliminate the confounding factor of chemical instability itself.

Aliquots of 247.5 μl hepatocytes were dispensed into each well of a 96-well uncoated plate. The plate was placed in an orbital shaker incubator for approximately 10 min. Aliquots of 2.5 μl of 100 μM test compounds were added to the appropriate wells of the uncoated 96-well plate to initiate the reaction. This assay was performed in duplicate. The plate was incubated in an orbital shaker incubator for the calculated time points. 20 μl of the contents were transferred and mixed with 6 volumes (120 μl) of cold acetonitrile with internal standard to stop the reaction at 5, 15, 30, 45, 60, 80, and 100 min. Samples were centrifuged for 20 min at 4000 g, and 100 µL aliquots of supernatants were used for LC-MS/MS analysis to measure test compounds.

In vitro hepatocyte clearance was estimated by determining the half-life (T _1/2 ) of the compounds from their initial concentrations. The ratios of the peak areas of each compound (test or control) to the IS (internal standard) were calculated. A curve of the dependence of Ln (% of control) on the incubation time (min) was plotted and the slope of the linear fitting line was calculated. The drug elimination rate constant k (min ^-1 ), T _1/2 (min) and internal clearance CL _int (μl/min/10 ⁶ cells) in vitro were calculated according to the following equations:

k= -slope coefficient

T _1/2 = 0.693/k

CL _int =k/C _hep

where C _hep (cells × µL ^-1 ) is the concentration of cells in the incubation system.

Methodology for determining Log D

10 µl of working solution from each cassette is placed in order into the corresponding position of the 96-well rack (Log D plate). 500 µl of saturated octanol are added to each well of the above Log D plate without a lid, followed by 500 µl of saturated phosphate buffer. Seal with a hermetically sealed PTFE/SIL lid for a 96-well plate.

The Log D plate is transferred to an Eppendorf Thermomixer Comfort plate shaker and shaken at 25°C, 2000 rpm for 2 hours.

The samples are centrifuged at 4000 rpm at 25°C for 30 minutes to separate the phases. A pipette and syringe are used to pipette approximately 100 µl of the octanol and buffer phases into a new 96-well plate, respectively.

5 µl of the octanol sample was transferred to a new 96-well plate, followed by the addition of 495 µl of a mixture of _H2O and acetonitrile containing the internal standard (1:1) as 100-fold diluted octanol samples. Mixing was carried out using a vortex mixer for 5 min at 1000 rpm.

50 µl of the 100-fold diluted samples were transferred to a new 96-well plate, followed by the addition of 450 µl of a mixture of _H2O and acetonitrile containing the internal standard (1:1) in the form of 1000-fold diluted octanol samples. The mixture was mixed using a vortex mixer for 5 min at 1000 rpm.

Octanol samples diluted 1000 times are successively diluted 10,000, 100,000, and 1,000,000 times with a mixture of _H2O and acetonitrile containing an internal standard (1:1).

50 µl of the buffer samples were transferred to a new 96-well plate, followed by the addition of 450 µl of a mixture of _H2O and acetonitrile containing the internal standard (1:1) as 10-fold dilutions of the buffer samples. Mixing was carried out using a vortex mixer for 5 minutes at 1000 rpm.

Tenfold diluted buffer samples were successively diluted 100, 1000 and 10000 times with a mixture of _H2O and acetonitrile containing an internal standard (1:1). Samples were assessed by LC/MS/MS analysis. All compounds were tested in single replicates.

All calculations are performed using Microsoft Excel. The concentrations of the test compound in the octanol/buffer solution are estimated by LC/MS/MS analysis. The Log D value of the test compound is calculated as follows:

DF stands for dilution factor.

Area Ratio Oct means the area ratio of octanol samples.

Area Ratio Buf means the ratio of the areas of the buffer samples.

A method for measuring protein binding in human plasma using equilibrium dialysis

Add 597 µl of control plasma to each well of a new plastic plate or separate plastic tube with 3 µl of working solution from each cassette and vortex at 1000 rpm for 5 minutes. The final volume percentage of organic solvent is 0.5% and the final concentration of test compound is 5 µM. Immediately transfer 50 µl of the spiked plasma suspension to a 96-well plate to act as a T=0 control. Process samples in the same manner as post-incubation samples. Place any remaining spiked plasma in the incubator for the duration of the assay.

Place the inserts open-end up into the wells of the main plate. Add 500 µl of phosphate buffer (pH 7.4) to the buffer chamber marked with a white ring. Add 300 µl of the spiked plasma sample to the sample chamber marked with a red ring. Cover the apparatus with a gas-permeable lid and incubate for 18 h at 37°C at 300 rpm with 5% CO ₂ on an orbital shaker in a CO ₂ incubator. At the end of the incubation, remove the lid and pipette 50 µl of the dialyzed samples from both the buffer and plasma chambers into a separate 96-well assay plate, respectively.

At the same time, the remaining spiked plasma sample in a plastic plate or a separate plastic tube is incubated for 18 hours at 37°C with 5% CO ₂ in a CO ₂ incubator. At time T, equal to 18 hours, 50 µl of the original spiked plasma suspension is transferred to a 96-well plate for analysis.

Add 50 µl human plasma to sample buffer and an equal volume of PBS (phosphate buffered saline) to the pooled plasma samples. Vortex the plate at 1000 rpm for 2 minutes and add 400 µl acetonitrile containing the appropriate internal standard (IS) to precipitate the protein and release the compound. Vortex at 1000 rpm for 10 minutes. Centrifuge for 30 minutes at 4000 rpm. Transfer 250 µl of the supernatant to new 96-well plates and centrifuge again (4000 rpm, 30 minutes). Then transfer 100 µl of the supernatant to new 96-well plates for analysis. Add 100 µl of distilled water to each sample and vortex for 5 min at 1000 rpm for LC-MS/MS analysis. All compounds were tested in single replicates at a concentration of 5 µM in human plasma.

All calculations are performed using Microsoft Excel

The percentage of unbound, the percentage of bound, and the recovery of the test compound are calculated as follows:

Results

Rapid oral absorption (SOA) assay in rats

The acute oral absorption (SOA) model is an in vivo screening model to detect compound penetration into the brain. To assess the ability of a compound to penetrate the blood-brain barrier in rats, the total brain/plasma ratio ( _Cr , _brain ) was measured using AUC _brain /AUC _plasma . The CSF (cerebrospinal fluid)/plasma ratio ( _Cr , _CSF ) was determined using either AUC _CSF /AUC _plasma or the mean CSF/ _plasma ratio at available time points after oral administration, respectively. Free fractions in the biological matrix were determined using in vitro binding assays in plasma and brain in a separate study. (1) _Cr,uu,brain and _Cr , _uu,CSF were calculated using the following equations:

Claims (63)

1. Compound of formula (I) where R ¹ is C _1-6 alkyl, where R ¹ may be substituted with one R ⁵ ; k is equal to 0; R ³ is halogen; n is 0-2; R ⁴ is a halogen; m is 0-2; A represents =N- or =C(R ⁶ )-; R ⁵ is hydroxy; R ⁶ is hydrogen; or a pharmaceutically acceptable salt thereof. 2. A compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, wherein R ¹ is methyl or hydroxymethyl. 3. A compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1 or 2, wherein R ³ is fluorine. 4. A compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 3, wherein R ⁴ is fluorine. 5. A compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 4, wherein A is =N-. 6. A compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5, selected from: (5-(8-amino-1-(4-phenoxyphenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol; (5-(8-amino-1-(2-fluoro-4-phenoxyphenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol; (5-(8-amino-1-(2,3-difluoro-4-phenoxyphenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol; (5-(8-amino-1-(4-(2,3-difluorophenoxy)phenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol; (5-(4-amino-5-(2-fluoro-4-phenoxyphenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-yl)methanol; (5-(4-amino-5-(2,3-difluoro-4-phenoxyphenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-yl)methanol; and (5-(4-amino-5-(4-(2,3-difluorophenoxy)phenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-yl)methanol. 7. A compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5, selected from: ((2R,5R)-5-(8-amino-1-(4-phenoxyphenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol; ((2R,5R)-5-(8-amino-1-(2-fluoro-4-phenoxyphenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol; ((2R,5R)-5-(8-amino-1-(2,3-difluoro-4-phenoxyphenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol; ((2R,5R)-5-(8-amino-1-(4-(2,3-difluorophenoxy)phenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol; ((2R,5R)-5-(4-amino-5-(2-fluoro-4-phenoxyphenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-yl)methanol; ((2R,5R)-5-(4-amino-5-(2,3-difluoro-4-phenoxyphenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-yl)methanol; ((2R,5R)-5-(4-amino-5-(4-(2,3-difluorophenoxy)phenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-yl)methanol; ((2S,5S)-5-(8-amino-1-(4-phenoxyphenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol; ((2S,5S)-5-(8-amino-1-(2-fluoro-4-phenoxyphenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol; ((2S,5S)-5-(8-amino-1-(2,3-difluoro-4-phenoxyphenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol; ((2S,5S)-5-(8-amino-1-(4-(2,3-difluorophenoxy)phenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol; ((2S,5S)-5-(4-amino-5-(2-fluoro-4-phenoxyphenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-yl)methanol; ((2S,5S)-5-(4-amino-5-(2,3-difluoro-4-phenoxyphenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-yl)methanol; ((2S,5S)-5-(4-amino-5-(4-(2,3-difluorophenoxy)phenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-yl)methanol; ((2S,5R)-5-(8-amino-1-(4-phenoxyphenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol; ((2S,5R)-5-(8-amino-1-(2-fluoro-4-phenoxyphenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol; ((2S,5R)-5-(8-amino-1-(2,3-difluoro-4-phenoxyphenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol; ((2S,5R)-5-(8-amino-1-(4-(2,3-difluorophenoxy)phenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol; ((2S,5R)-5-(4-amino-5-(2-fluoro-4-phenoxyphenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-yl)methanol; ((2S,5R)-5-(4-amino-5-(2,3-difluoro-4-phenoxyphenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-yl)methanol; ((2S,5R)-5-(4-amino-5-(4-(2,3-difluorophenoxy)phenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-yl)methanol; ((2R,5S)-5-(8-amino-1-(4-phenoxyphenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol; ((2R,5S)-5-(8-amino-1-(2-fluoro-4-phenoxyphenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol; ((2R,5S)-5-(8-amino-1-(2,3-difluoro-4-phenoxyphenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol; ((2R,5S)-5-(8-amino-1-(4-(2,3-difluorophenoxy)phenyl)imidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methanol; ((2R,5S)-5-(4-amino-5-(2-fluoro-4-phenoxyphenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-yl)methanol; ((2R,5S)-5-(4-amino-5-(2,3-difluoro-4-phenoxyphenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-yl)methanol; and ((2R,5S)-5-(4-amino-5-(4-(2,3-difluorophenoxy)phenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)tetrahydro-2H-pyran-2-yl)methanol. 8. A pharmaceutical composition for inhibiting Bruton's tyrosine kinase (BTK), comprising an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 7 in combination with a pharmaceutically acceptable diluent or carrier. 9. A compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 7 for use as a medicament for inhibiting BTK. 10. A method for treating diseases or disorders mediated in whole or in part by BTK in a warm-blooded animal, comprising administering to said animal an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 7, wherein said disease or disorder is selected from immune disorders, cancer, cardiovascular diseases, viral infections, metabolic/endocrine disorders, neurological disorders, allergic diseases, autoimmune diseases and inflammatory diseases. 11. The method of claim 10, wherein the cancer is selected from small lymphocytic lymphoma (SLL), follicular lymphoma, Richter's transformation, mantle cell lymphoma, chronic lymphocytic leukemia (CLL), Waldenstrom macroglobulinemia, non-Hodgkin's lymphoma, primary central nervous system lymphoma, secondary central nervous system lymphoma, or diffuse large B-cell lymphoma. 12. The method of claim 10, wherein the disease or disorder is selected from urticaria/Sjogren's syndrome, rheumatoid arthritis, osteoporosis, vasculitis, idiopathic thrombocytopenic purpura (ITP), severe myasthenia, allergic rhinitis, asthma, multiple sclerosis, and systemic lupus erythematosus. 13. The method according to claim 10, wherein the warm-blooded animal is a human. 14. The method of claim 12, wherein the disease or disorder is multiple sclerosis. 15. A method for inhibiting BTK comprising administering to an animal an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1-7. 16. The method of claim 15, wherein the BTK is wild-type BTK or BTK with the C481 mutation. 17. The method according to claim 16, wherein BTK with mutation C481 is selected from the group consisting of BTK with mutation C481S, BTK with mutation C481Y, BTK with mutation C481R and BTK with mutation C481F.