WO2012035055A1 - Novel compounds - Google Patents

Novel compounds Download PDF

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Publication number
WO2012035055A1
WO2012035055A1 PCT/EP2011/065921 EP2011065921W WO2012035055A1 WO 2012035055 A1 WO2012035055 A1 WO 2012035055A1 EP 2011065921 W EP2011065921 W EP 2011065921W WO 2012035055 A1 WO2012035055 A1 WO 2012035055A1
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Prior art keywords
pyridin
amino
thiazolo
methyl
pyridinediamine
Prior art date
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PCT/EP2011/065921
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French (fr)
Inventor
Nicholas Paul Barton
Sebastien Andre Campos
Robin Arthur Carr
John David Harling
Ian Edward David Smith
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Glaxo Group Limited
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Publication of WO2012035055A1 publication Critical patent/WO2012035055A1/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D513/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for in groups C07D463/00, C07D477/00 or C07D499/00 - C07D507/00
    • C07D513/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for in groups C07D463/00, C07D477/00 or C07D499/00 - C07D507/00 in which the condensed system contains two hetero rings
    • C07D513/04—Ortho-condensed systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/445—Non condensed piperidines, e.g. piperocaine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/496—Non-condensed piperazines containing further heterocyclic rings, e.g. rifampin, thiothixene or sparfloxacin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/506—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim not condensed and containing further heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
    • A61K31/5375—1,4-Oxazines, e.g. morpholine
    • A61K31/5377—1,4-Oxazines, e.g. morpholine not condensed and containing further heterocyclic rings, e.g. timolol
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
    • C07D417/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing three or more hetero rings

Definitions

  • the present invention is directed to certain novel compounds which are inhibitors of kinase activity, processes for their preparation, pharmaceutical compositions comprising the compounds, and the use of the compounds or the compositions in the treatment of various disorders. More specifically, the compounds of the invention are inhibitors of the activity or function of Itk (interleukin-2 inducible tyrosine kinase).
  • Itk interleukin-2 inducible tyrosine kinase
  • Compounds which are inhibitors of the activity or function of Itk may be useful in the treatment of disorders such as respiratory diseases including asthma, chronic obstructive pulmonary disease (COPD) and bronchitis; allergic diseases including allergic rhinitis and atopic dermatitis; autoimmune diseases including rheumatoid arthritis, multiple sclerosis, psoriasis, type I diabetes, T cell mediated hypersensitivities, Guiliain-Barre Syndrome and Hashimoto's thyroiditis; transplant rejection; graft versus host disease; inflammatory disorders including conjunctivitis, contact dermatitis, inflammatory bowel disease and chronic inflammation; HIV; aplastic anemia; and pain including inflammatory pain.
  • respiratory diseases including asthma, chronic obstructive pulmonary disease (COPD) and bronchitis
  • allergic diseases including allergic rhinitis and atopic dermatitis
  • autoimmune diseases including rheumatoid arthritis, multiple sclerosis, psoriasis,
  • lnterleukin-2 inducible tyrosine kinase is a non-receptor tyrosine kinase of the Tec family, which is also known as Tsk or Emt.
  • Other members of the Tec fami!iy include: Tec, Btk, Txk and Bmx.
  • the Tec family kinases are predominantly expressed in haematopoietic cells, however Bmx and Tec have a wider expression profile.
  • the Tec famiiy kinases share a common domain structure: an amino-terminal pleckstrin homology (PH) domain (absent in Txk), a tec homology domain (containing one or two proline rich regions), followed by Src homology SH3 and SH2 domains, and a carboxy-terminal kinase domain.
  • the PH domain binds to Ptdln(3,4,5)P3, and is responsible for locating the Tec kinase to the plasma membrane, whilst the PRR, SH3 and SH2 domains are involved in protein-interactions important in formation of the signalling complex.
  • Itk expression is restricted to T cells, NK and mast cells. Itk is the predominant Tec family kinase in naive T cells, which also express Txk and Tec. Upon activation via the T cell receptor or interleukin-2 (IL-2), the expression of Itk increases. There is some evidence that Itk is preferentially expressed in Th2 over Th1 ceils, in contrast to Txk which is present at higher levels in Th1 cells (1 ). Itk plays a key role in T cell receptor signalling. Itk is recruited to the plasma membrane through interaction with Ptdlns(3,4,5)P3, which is generated by P!3kinase. Itk forms a complex with several signalling and scaffold proteins including SLP76 and LAT.
  • Itk is transphorphorylated by Lck. Activated Itk phosphoryiates PLCy, leading to the generation of lns(1 ,4,5)P3 (required for calcium flux within the cells) and diacylglycerol (activates members of the protein kinase C family and RAS guanyl-releasing protein. This results in the activation of mitogen-activated protein kinases (including JNK and ERK) and other effectors that regulate gene transcription, leading to the secretion of cytokines (reviewed in ref 2). In addition to the role of itk in PLCy activation and Ca 2+ mobilisation, Itk may also contribute to TCR-induced actin reorganisation, and formation of the immune synapse.
  • actin cytoskeleton may not require kinase activity (3), suggesting the importance of Itk as a scaffold protein.
  • itk may also be activated via the chemokine receptor CXCR4 (4) in T cells, and via the FceRI in mast cells (5).
  • CsA Cyclosporin A
  • CsA inhibited the late phase but not the early phase response (9), suggesting that effects on mast cells are unlikely to play a key role in the beneficial effect seen of CsA.
  • daclizumab an antibody against the anti-IL-2Ra chain (CD25) of activated lymphocytes improved pulmonary function and asthma control in patients with moderate to severe chronic asthma (10), supporting anti-T cell therapy for asthma.
  • Inhibition of Itk represents a potential novel therapy for asthma, by inhibiting T cell cytokine release.
  • the key role for Itk in T cell receptor signalling has been demonstrated using Itk-/- mice and siRNA.
  • In vitro activation of CD4+ cells from Itk knockout mice show reduced levels of Th2 (11) or both Th1 and Th2 (12) cytokines compared to wild type.
  • Naive T cells from Itk knockout mice can differentiate normally into either Th1 or Th2 cells if cultured in vitro under appropriate cytokine conditions, suggesting that Itk is not required for Th2 cell differentiation (12).
  • Itk may be beneficial in a variety of T-ce!l mediated diseases.
  • Itk may play a role in other allergic diseases such as allergic rhinitis and atopic dermatitis.
  • Single nucleotide polymorphisms in Itk have been associated with atopy (18) and seasonal ailergic rhinitis (19).
  • itk mRNA levels in the peripheral blood T cells of atopic dermatitis patient is elevated in T cells from affected patients, compared to healthy controls (20).
  • Such disorders include respiratory diseases including asthma, chronic obstructive pulmonary disease (COPD) and bronchitis; allergic diseases including allergic rhinitis and atopic dermatitis; autoimmune diseases including rheumatoid arthritis, multiple sclerosis, psoriasis, type I diabetes, T cell mediated hypersensitivities, Guil!ain-Barre Syndrome and Hashimoto's thyroiditis; transplant rejection; graft versus host disease; inflammatory disorders including conjunctivitis, contact dermatitis, inflammatory bowel disease and chronic inflammation; HIV; aplastic anemia; and pain including inflammatory pain.
  • respiratory diseases including asthma, chronic obstructive pulmonary disease (COPD) and bronchitis
  • allergic diseases including allergic rhinitis and atopic dermatitis
  • autoimmune diseases including rheumatoid arthritis, multiple sclerosis, psoriasis, type I diabetes, T cell mediated hypersensitivities, Guil!a
  • compounds of the invention may show selectivity for itk over other kinases.
  • compounds of the invention may be potent inhibitors of Itk activity.
  • compounds of the invention may be irreversible inhibitors of Itk activity.
  • compounds of the invention may show a fast rate of irreversible inhibition of Itk activity.
  • the invention is directed to certain novel compounds. Specifically, the invention is directed to compounds of formula (i):
  • R 1 to R 3 , X and Y are as defined below, and salts thereof.
  • the compounds are inhibitors of kinase activity, in particular Itk activity, and may have the potential for irreversible inhibition. Compounds which are Itk inhibitors may be useful in the treatment of disorders associated with inappropriate Itk activity, such as asthma. Accordingly, the invention is further directed to pharmaceutical compositions comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof. The invention is still further directed to methods of inhibiting Itk activity and treatment of disorders associated therewith using a compound of formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising a compound of formula (I) or a pharmaceuticaliy acceptable salt thereof. The invention is yet further directed towards processes for the preparation of the compounds of the invention.
  • Figure 1 shows an X-ray powder diffraction (XRPD) pattern for A/-[(3S)-1 -acryloyl-3- pyrrolidinyl]-W-[1 ,3]thiazolo[5,4-b]pyridin-2-yl-4-( ⁇ [(1 S)-1 ,2,2- trimethylpropyl]amino ⁇ methyi)-2,6-pyridinediamine difumarate monohydrate.
  • XRPD X-ray powder diffraction
  • the invention is directed to compounds of formula (I)
  • R 1 is hydrogen, -CH 2 OR 4 , -CH 2 NR 5 R 6 , -CH 2 -pheny[ wherein ihe phenyl is optionally substituted by one or two substituents independently selected from C h alky!, C 1-6 alkoxy, nitrile and halo, -CH 2 -6-membered heterocyclyl wherein the 6-membered heterocyclyl optionally contains an oxygen atom and is optionally substituted by one or two substituents independently selected from Ci assign 6 aSkyl and halo, or -CH 2 -5-membered heteroaryl wherein the 5-membered heteroaryl contains one or two nitrogen atoms and is optionally substituted by C 1-6 alkyl;
  • R 2 is -(NH) 2 COCHCH 2> -NR 7 R 8 , -OR 9 or phenyl wherein the phenyl is optionally substituted by -NR 10 R 1 1 ;
  • R 3 is hydrogen, halo, C-
  • R 6 and R 6 are each independently hydrogen or C ⁇ alkyl, or
  • R 5 and R 6 together with the nitrogen atom to which they are attached, are linked to form a
  • 5- or 6-membered heterocyclyl wherein the 6-membered heterocyclyl optionally contains an oxygen atom or a further nitrogen atom and the 5- or 6-membered heterocyclyl is optionally substituted by one or two substituents independently selected from C 1-6 alky! and halo;
  • R 7 is hydrogen or methyl
  • R 8 is -COCHCH 2l - ⁇ CH 2 ) n NR 12 R 13 , C 3 . 6 cycloalkyl substituted by -OS0 2 CH 3 or -NR 14 R 1S , or 4-, 5- or 6-membered heterocyclyl wherein the 4-, 5- or 6-membered heterocyclyl contains a nitrogen atom and is substituted by -R 16 , or
  • R 7 and R 8 together with the nitrogen atom to which they are attached, are linked to form a
  • 6- membered heterocyclyl optionally containing a further nitrogen atom wherein the 6- membered heterocyclyl is substituted by -R 17 or -NR 18 R 19 ;
  • R 9 is 5- or 6-membered heterocyclyl wherein the 5- or 6-membered heterocyclyl contains a nitrogen atom and is substituted by -R 20 ;
  • R 1 1 , R 13 , R 15 , R 6 , R 17 , R 19 and R 20 are each independently -COCR 25 CR 21 R 26 , -COCH 2 halo or -COCH 2 OR 22 ;
  • R 2 is hydrogen, methyl, -CF 3 or -CH 2 NR 23 R 24 ;
  • R 22 is phenyl optionally substituted by halo;
  • R 4 , R 10 , R 2 , R 14 , R 18 , R 23 and R 24 are each independently hydrogen or C h alky!;
  • R 25 is hydrogen, methyl or -CH 2 NR 27 R 28 ;
  • R 26 is hydrogen or methyl; and R , together with the nitrogen atom to which they are attached, are linked to form a 6-membered heterocyclyl optional!y containing an oxygen atom; X and Y are each independently -N- or -CH-; and n is 2, 3 or 4; and salts thereof (hereinafter “compounds of the invention").
  • R 2 is -(NH) 2 COCHCH 2
  • R 8 is - COCHCH 2l
  • R 11 , R 13 , R 5 , R 16 , R 7 , R 19 and R 20 are each independently - COCR 25 CR 21 R 26 , contain carbon-carbon double bonds and may be depicted as shown below wherein "-" denotes the point of attachment:
  • R is hydrogen, ⁇ CH 2 OR 4 , -CH 2 NR 5 R S , -CH 2 -phenyl wherein the phenyl is optionally substituted by one or two substituents independently selected from 6 alkyl, C 6 a!koxy, nitrilo and halo, or -CH 2 -5-membered heteroaryl wherein the 5- membered heteroaryl contains one or two nitrogen atoms and is optionally substituted by C 1-6 alkyl.
  • R 1 is hydrogen, -CH 2 OR 4 , -CH 2 NR 5 R 6 or -CH 2 -5- membered heteroaryl wherein the 5-membered heteroaryl contains one or two nitrogen atoms and is optionally substituted by C h alky!.
  • R 1 is hydrogen, - CH 2 OR 4 , -CH 2 -6-membered heterocyclyl wherein the 6-membered heterocyclyl optionally contains an oxygen atom and is optionally substituted by one or two substituents independently selected from C h alky! and halo, -CH 2 -5-membered heteroaryi wherein the heteroaryi contains one or two nitrogen atoms and is optionally substituted by C h alky!.
  • R 1 is -Ch ⁇ OR 4 , or -CH 2 -6-membered heterocyciyl wherein the 6- membered heterocyciyl optionally contains an oxygen atom and is optionally substituted by one or two substituents independently selected from C 1-6 alkyl and halo.
  • R 1 is -CH 2 OR 4 , in particular wherein R 4 is methyl, !n another embodiment, R 1 is -CH 2 -6-membered heterocyciyl wherein the 6-membered heterocyciyl optionally contains an oxygen atom and is optionally substituted by one or two substituents independently selected from C 1-6 alky! and halo, in particular N-morpho!inyl.
  • R 1 is -CH 2 NR 5 R 6 .
  • R 2 is -NR 7 R 8 , -OR 9 or phenyl wherein the phenyl is optionally substituted by -NR 10 R 11 .
  • R 2 is -NR 7 R 8 .
  • R 2 is -NR 7 R 8 wherein R 7 is hydrogen. In one embodiment, R 2 is -NR 7 R 8 wherein R 8 is -(CH 2 ) n NR 12 R 13 .
  • R 2 is -NR 7 R 8 wherein R 8 is C 3 . 6 cycloalkyl substituted by -NR 14 R 15 , in particular C 6 cycloalkyl substituted by -NR 14 R 15 . In one embodiment, R 2 is -NR 7 R 8 wherein R 8 is 5- or 6-membered heterocyciyl wherein the 5- or 6-membered heterocyciyl contains a nitrogen atom and is substituted by -R 16 .
  • R 2 is -NR 7 R 8 wherein R 8 is 5-membered heterocyciyl wherein the 5- contains a nitrogen atom and is substituted by -R 16 .
  • R 12 or R 14 are each independently hydrogen.
  • R 3 is hydrogen or halo, for example bromo. In another embodiment, R 3 is hydrogen. In a further embodiment, R 3 is C 1-6 alkoxy, for example methoxy. in one embodiment, R 4 is C h alky!, for example methyl.
  • R s and R 6 are each independently hydrogen or C h alky!.
  • R 5 is hydrogen and R e is C 1-6 alkyi, for example 3,3-dimethylbutan-2-yl- (1 ,2,2-trimethylpropyl-).
  • R 5 and R 6 are each independently C-,. 6 alkyl, for example methyl.
  • R 5 and R 6 together with the nitrogen atom to which they are attached, are linked to form a 5- or 6-membered heterocyciyi wherein the 6-membered heterocyciyi optionally contains an oxygen atom or a further nitrogen atom and the 5- or 6- membered heterocyciyi is optionally substituted by one or two substituents independently selected from C -6 alkyl and halo.
  • R 5 and R 6 together with the nitrogen atom to which they are attached, are linked to form a 5- or 6-membered heterocyciyi wherein the 6-membered heterocyciyi optionally contains an oxygen atom or a further nitrogen atom and the 5- or 6-membered heterocyciyi is optionally substituted by one or two substituents independently selected from d-ealkyl.
  • R 5 and R 6 together with the nitrogen atom to which they are attached, are linked to form a 6- membered heterocyciyi wherein the 6-membered heterocyciyi optionally contains an oxygen atom and is optionally substituted by one or two substituents independently selected from C ⁇ alkyl and halo.
  • R 5 and R 6 together with the nitrogen atom to which they are attached, are linked to form a 6-membered heterocyciyi wherein the 6-membered heterocyciyi contains an oxygen atom and is substituted by two substituents independentiy selected from C h alky!, for example methyi.
  • R 7 is hydrogen or methyl
  • R 8 is -COCHCH 2 , -(CH 2 ) n NR 12 R 13 , C 3 . 6 cycloalkyi substituted by -OS0 2 CH 3 or -NR 1 "R 15 , or 5- or 6-membered heterocyciyi wherein the 5- or 6-membered heterocyciyi contains a nitrogen atom and is substituted by -R 16 , or R 7 and R 8 , together with the nitrogen atom to which they are attached, are linked to form a 6-membered heterocyclyl optionally containing a further nitrogen atom wherein the 6- membered heterocyclyl is substituted by -R 7 or -NR 18 R 19 .
  • R 7 is hydrogen.
  • R 8 is 5- or 6-membered heterocyclyl wherein the 5- or 6-membered heterocyclyl contains a nitrogen atom and is substituted by -R 16 . In a further embodiment, R 8 is 5-membered heterocyclyl wherein the 5-membered heterocyclyl contains a nitrogen atom and is substituted by -R 16 .
  • R 9 is 6-membered heterocyclyl wherein the 6-membered heterocyclyl contains a nitrogen atom and is substituted by -R 20 .
  • R 10 is hydrogen.
  • R 1 ⁇ R 13 , R 15 , R 16 , R 17 , R 19 and R 20 are each independently - COCHCHR 21 , -COCH 2 halo or -COCH 2 OR 22 . In one embodiment, R 11 -COCHCHR 21 .
  • R 12 is hydrogen. In a further embodiment, R 12 is Ci. 6 alkyl, for example methyl. In one embodiment, R 3 is -COCHCHR 21 . in one embodiment, R 4 is hydrogen.
  • R 15 is -COCHCHR 21 .
  • R 16 is -COCHCHR 21 , -COCH 2 halo or -COCH 2 OR 2Z . In a further embodiment, R 16 is -COCHCHR 21 .
  • R 7 is -COCHCHR 1 in one embodiment, R 1s is hydrogen. In one embodiment, R 19 is -COCHCHR 21 .
  • R 20 is -COCHCHR 21 .
  • R 21 is hydrogen, -CF 3 or -CH 2 NR 23 R 24 .
  • R 21 is hydrogen.
  • R 22 is phenyl optionally substituted by fluoro.
  • R 23 is C 1-6 aikyl, for example methyl.
  • R 24 is C ⁇ alkyl, for example methyl.
  • R 25 is hydrogen.
  • R 26 is hydrogen
  • R 27 and R 28 together with the nitrogen atom to which they are attached, are linked to form morpho!inyl.
  • X is -N-. In a further embodiment, X is -CH-.
  • Y is -N-. In a further embodiment, Y is -CH-. In one embodiment, n is 2.
  • R 1 is hydrogen, -CH 2 OR 4 , -CH 2 NR 5 R 6 , -CH 2 -phenyl wherein the phenyl is optionally substituted by one or two substituents independently selected from C h alky! , C ⁇ alkoxy, nitrilo and halo, -CH r 6-membered heterocyciyi wherein the 6-membered heterocydyi optionally contains an oxygen atom and is optionally substituted by one or two substituents independently selected from C ⁇ alkyl and halo, or -CH 2 -5-membered heteroary! wherein the 5-membered heteroaryl contains one or two nitrogen atoms and is optionally substituted by Chalky!;
  • R 2 is -(NH) 2 COCHCH 2 , -NR 7 R 8 , -OR 9 or phenyl wherein the phenyl is optionally substituted by -NR 10 R 11 ;
  • R 3 is hydrogen, halo, Chalky!, C -6 alkoxy;
  • R 5 and R 6 are each independently hydrogen or C 1-6 aikyl, or
  • R 5 and R 6 together with the nitrogen atom to which they are attached, are linked to form a 5- or 6-membered heterocydyi wherein the 6-membered heterocyciyi optionally contains an oxygen atom or a further nitrogen atom and the 5- or 6-membered heterocydyi is optionally substituted by one or two substituents independently selected from C h alky! and halo;
  • R 7 is hydrogen or methyl
  • R 8 is -COCHCH 2] ⁇ (CH 2 ) n NR 1 R 13 , C 3-6 cycloalkyi substituted by -OS0 2 CH 3 or -NR 1 R 5 , or
  • R 7 and R 8 together with the nitrogen atom to which they are attached, are linked to form a
  • 6- membered heterocydyi optionally containing a further nitrogen atom wherein the 6- membered heterocyciyi is substituted by -R 17 or -NR 18 R 19 ;
  • R 9 is 5- or 6-membered heterocyciyi wherein the 5- or 6-membered heterocyciyi contains a nitrogen atom and is substituted by -R 20 ;
  • R 11 , R 13 , R 5 , R 16 , R 17 , R 19 and R 20 are each independently -COCHCHR 21 , -COCH 2 haio or - COCH 2 OR 22 ;
  • R 21 is hydrogen, -CF 3 or -CH 2 NR 23 R 24 ;
  • R 22 is phenyl optionally substituted by halo;
  • R 4 , R 10 , R 12 , R 14 , R 8 , R 23 and R 24 are each independently hydrogen or C h alky!;
  • X and Y are each independently -N- or -CH-; and n is 2, 3 or 4; and salts thereof.
  • the invention is directed to compounds of formula (IB)
  • R 1 is hydrogen, -CH 2 OR 4 , -CH 2 NR 5 R 6 , -CH 2 -phenyl wherein the phenyl is optionally substituted by one or two substituents independently selected from Ci -6 alkyl, C 1-6 alkoxy, nstrilo and halo, -CH 2 -6-membered heterocyclyl wherein the 6-membered heterocyclyl optionally contains an oxygen atom and is optionally substituted by one or two substituents independently selected from C 1-6 alkyl and halo, or -CH 2 -5-membered heteroaryl wherein the 5-membered heteroaryl contains one or two nitrogen atoms and is optionally substituted by C 1-6 alkyl;
  • R 2 is -NR 7 R 8 , -OR 9 or phenyl wherein the phenyl is optionally substituted by -NR 10 R 1 1 ;
  • R 3 is hydrogen, halo, C 1-6 alkyl, C ⁇ alkoxy;
  • R 5 and R 6 together with the nitrogen atom to which they are attached, are linked to form a 6-membered heterocycly! wherein the 6-membered heterocyclyl optionally contains an oxygen atom and is optionaliy substituted by one or two substituents independently selected from C ⁇ alkyl and halo;
  • R 7 is hydrogen or methyl
  • R 8 is -(CH 2 ) n NR 12 R 13 , C 3 . 6 cycloalkyl substituted by -OS0 2 CH 3 or -NR 14 R 15 , or 5- or 6- membered heterocyciyi wherein the 5- or 6-membered heterocyciyi contains a nitrogen atom and is substituted by -R 16 , or
  • R 7 and R 8 together with the nitrogen atom to which they are attached, are linked to form a 6-membered heterocyciyi optionally containing a further nitrogen atom wherein the 6- membered heterocyciyi is substituted by -R 17 or -NR 18 R 19 ;
  • R 9 is 5- or 6-membered heterocycly! wherein the 5- or 6-membered heterocyciyi contains a nitrogen atom and is substituted by -R 20 ;
  • R 1 , R 13 , R 15 , R 16 , R 17 , R 19 and R 20 are each independently -COCHCHR 21 , -COCH 2 halo or - COCH 2 OR 22 ;
  • R 21 is hydrogen, -CF 3 or -CH 2 NR 23 R 24 ;
  • R 22 is phenyl optionally substituted by halo
  • R 4 , R 1Q , R 12 , R 14 , R 18 , R 23 and R 24 are each independently hydrogen or C ⁇ alkyl; X and Y are each independently -N- or -CH-; and n is 2, 3 or 4; and salts thereof.
  • the invention is directed to compounds of formula (IC)
  • R is -CH 2 NR 5 R 6 ;
  • R 2 is -NR 7 R 8 ;
  • R 3 is hydrogen
  • R 5 and R 6 are each independently hydrogen or C 1-6 alkyl, or
  • R 5 and R 6 together with the nitrogen atom to which they are attached, are linked to form a 6-membered heterocyclyl wherein the 6-membered heterocyclyl contains an oxygen atom and is substituted by two substituents independently selected from C 1-e alkyl;
  • R 7 is hydrogen
  • R 8 is 5-membered heterocyclyl wherein the 5-membered heterocyclyl contains a nitrogen atom and is substituted by -R 6 ;
  • R 16 is -COCHCHR 21 ;
  • R 21 is hydrogen;
  • X is -N-; and
  • Y is -CH-; and salts thereof.
  • R 1 is -CH 2 NR 5 R 6 ;
  • R 2 is -NR 7 R e ;
  • R 3 is hydrogen;
  • R 5 and R 6 are each independently hydrogen or C 1-6 alkyl;
  • R 7 is hydrogen;
  • R 8 is 5-membered heterocyciyi wherein the 5-membered heterocyciyi contains a nitrogen atom and is substituted by -R 16 ;
  • R 6 is -COCHCHR 21 ;
  • R 21 is hydrogen;
  • X is -N-
  • Y is -CH-; and salts thereof.
  • Compounds of the invention include the compounds of Examples 1 to 102 and salts thereof.
  • the compound of the invention is selected from the group consisting of:
  • the compound of the invention is selected from the group consisting of:
  • the compound of the invention is selected from the group consisting of:
  • the compound of the invention is selected from the group consisting of:
  • the compound of the invention is selected from:
  • the compound of the invention is:
  • the compound of the invention is:
  • the compound of the invention is:
  • the compound of the invention is:
  • the compound of formula (I) or a salt thereof is selected from the group consisting of:
  • AlkyI refers to a saturated hydrocarbon chain having the specified number of member atoms.
  • C 1-6 alkyl refers to an alky! group having from 1 to 6 member atoms, for example 1 to 4 member atoms.
  • AlkyI groups may be optionally substituted with one or more substituents as defined herein.
  • AlkyI groups may be straight or branched. Representative branched alkyl groups have one, two, or three branches.
  • AlkyI includes methyl, ethyl, propyl (n-propyl and isopropyl), butyl (n-butyl, isobutyl, and t-butyl), pentyl (n-pentyl, isopentyl, and neopentyl), and hexyl.
  • Alkyl groups may also be part of other groups, for example alkoxy.
  • Alkynyl refers to a hydrocarbon chain having the specified number of member atoms and at least one triple bond.
  • C 2 . 6 alkynyl refers to an alkynyl group having from 2 to 6 member atoms, for example 2 to 4 member atoms.
  • Alkynyl groups may be straight or branched.
  • Alkynyl includes ethynyl, 1-propynyl, 1 -butynyl, 2-butynyl, 1- pentynyi, 2-pentynyl, 3-pentynyl, 1-hexynyl, 2-hexynyi and 3-hexynyl.
  • Cycloalkyl refers to a saturated hydrocarbon ring having the specified number of member atoms. Cycloalkyl groups are monocyclic ring systems. For example, C 3 . ecycloalkyl refers to a cycloalkyl group having from 3 to 6 member atoms. Cycloalkyl groups may be optionally substituted with one or more substituents as defined herein. Cycloalkyl includes cyc!opropyl, cyc!obutyl, cyclopentyi, and cyc!ohexyi. In one embodiment, the cycloalkyl groups herein are cyclohexyl.
  • Enantiomerically enriched refers to products whose enantiomeric excess is greater than zero, For example, enantiomerically enriched refers to products whose enantiomeric excess is greater than 50% ee, greater than 75% ee, and greater than 90% ee.
  • Enantiomeric excess or "ee” is the excess of one enantiomer over the other expressed as a percentage. As a result, since both enantiomers are present in equal amounts in a racemic mixture, the enantiomeric excess is zero (0% ee). However, if one enantiomer was enriched such that it constitutes 95% of the product, then the enantiomeric excess would be 90% ee (the amount of the enriched enantiomer, 95%, minus the amount of the other enantiomer, 5%). “Enantiomerically pure” refers to products whose enantiomeric excess is 99% ee or greater. “Half-life” (or “half-lives”) refers to the time required for half of a quantity of a substance to be converted to another chemically distinct species in vitro or in vivo.
  • Halo refers to the halogen radical fluoro, chloro, bromo, or iodo.
  • Heteroaryl refers to an aromatic ring containing from 1 to 3 heteroatoms as member atoms in the ring or rings. Heteroaryl groups containing more than one heteroatom may contain different heteroatoms. Heteroaryl groups may be optionally substituted with one or more substituents if so defined herein.
  • the heteroaryl groups herein are monocyclic ring systems or are fused bicyclic ring systems. Monocyclic heteroaryl rings have 5 or 6 member atoms. Bicyclic heteroaryl rings have 9 or 10 member atoms.
  • Monocyclic heteroaryl includes pyrrolyl, furany!, pyrazolyl, imidazo!yl, oxazolyl, isoxazolyl, oxadiazoiyl, triazolyl, pyridinyl, pyrimidinyl, pyridazinyl and pyrazinyi.
  • Bicyclic heteroaryl includes indolyl, isoindolyl, indolizinyl, indazolyl, benzimidazoly!, pyrrolopyridinyl, quinoiinyl, isoquinoiinyl, quinoxalinyl, quinazolinyl and naphthridinyi.
  • the heteroaryl groups herein are monocyclic ring systems having 5 member atoms, for example pyrrolyl, pyrazolyl or imidazolyl.
  • Heteroatom refers to a nitrogen, sulphur, or oxygen atom.
  • Heterocyclyl refers to a saturated or unsaturated ring containing 1 or 2 heteroatoms as member atoms in the ring. However, heterocyclyl rings are not aromatic. In certain embodiments, heterocyclyl is saturated. In other embodiments, heterocyclyl is unsaturated but not aromatic. Heterocyclyl groups containing more than one heteroatom may contain different heteroatoms.
  • the heterocyclyl groups herein are monocyclic ring systems having 4, 5 or 6 member atoms. Heterocyclyl groups may be optionally substituted with one or more substituents as defined herein.
  • Heterocyclyl includes azetidinyl, pyrrolidinyl, pyrazo!idinyl, imidazolinyl, oxazoiidiny!, isoxazolidinyl, piperidinyl, piperazinyl and morphoiinyl.
  • the heterocyclyl groups herein are monocyclic ring systems having 5 or 6 member atoms, for example pyrrolidinyl, piperidinyl, piperazinyl, tetrahydro-1 ⁇ 2H)-pyridazinyl and morphoiinyl.
  • Member atoms refers to the atom or atoms that form a chain or ring. Where more than one member atom is present in a chain and within a ring, each member atom is covalently bound to an adjacent member atom in the chain or ring. Atoms that make up a substituent group on a chain or ring are not member atoms in the chain or ring. "Optionally substituted” indicates that a group, such as heteroaryl, may be unsubstituted or substituted with one or more substituents as defined herein. "Substituted” in reference to a group indicates that a hydrogen atom attached to a member atom within a group is replaced.
  • substituted includes the implicit provision that such substitution be in accordance with the permitted valence of the substituted atom and the substituent and that the substitution results in a stable compound (i.e. one that does not spontaneously undergo transformation such as by rearrangement, cyclization, or elimination).
  • a single atom may be substituted with more than one substituent as long as such substitution is in accordance with the permitted valence of the atom.
  • Suitable substituents are defined herein for each substituted or optionally substituted group.
  • “Pharmaceutically acceptable” refers to those compounds, materials, compositions, and dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, or other problem or complication, commensurate with a reasonable benefit/risk ratio.
  • the compounds of the invention may exist in solid or liquid form. In the solid state, the compounds of the invention may exist in crystalline or noncrystalline form, or as a mixture thereof.
  • pharmaceutically acceptable solvates may be formed wherein solvent molecules are incorporated into the crystalline lattice during crystallization. Solvates may involve nonaqueous solvents such as ethanol, isopropanoi, DMSO, acetic acid, ethano!amine, and EtOAc, or they may involve water as the solvent that is incorporated into the crystalline lattice.
  • Solvates wherein water is the solvent that is incorporated into the crystalline lattice are typically referred to as "hydrates.” Hydrates include stoichiometric hydrates as well as compositions containing variable amounts of water. The invention includes ail such solvates.
  • the compound of the invention is /V-[ ⁇ 3S)-1-acryloyl-3-pyrrolidinyl]-/V-[1 ,3]thiazolo[5,4-&]pyridin-2-yl-4-( ⁇ [(1 S)- 1 ,2,2-trimethyipropyl]amino ⁇ methy[)-2,6-pyridinediamine difumarate monohydrate.
  • polymorphs may exhibit poiymorphism (i.e. the capacity to occur in different crystalline structures). These different crystalline forms are typically known as "polymorphs".
  • the invention includes all such polymorphs. Polymorphs have the same chemical composition but differ in packing, geometrical arrangement, and other descriptive properties of the crystalline solid state. Polymorphs, therefore, may have different physical properties such as shape, density, hardness, deformabi!ity, stability, and dissolution properties. Polymorphs typically exhibit different melting points, IR spectra, and X-ray powder diffraction patterns, which may be used for identification.
  • polymorphs may be produced, for example, by changing or adjusting the reaction conditions or reagents, used in making the compound. For example, changes in temperature, pressure, or solvent may result in polymorphs. In addition, one polymorph may spontaneously convert to another polymorph under certain conditions.
  • the present invention provides W-[(3S)-1-acryloyl-3-pyrrolidinyl]-W- [1 ,3]thiazoio[5,4-b]pyridin-2-yl-4-( ⁇ [(1 S)-1 ,2,2-trimethylpropyi]amino ⁇ methyl ⁇ -2,6- pyridinediamine or a pharmaceutically acceptable salt thereof in crystalline form.
  • the present invention provides A/-[(3S)-1-acryloyl-3-pyrrolidinyl]- /V-[1 ,3]thiazolo[5,4-/b]pyridin-2-yl-4-( ⁇ [(1 S)-1 ,2,2-trimethyipropyl]amino ⁇ methyi)-2,6- pyridinediamine difumarate monohydrate in crystalline form.
  • the present invention provides a polymorph of /v/-[(3S)-1 ⁇ acryloyl- 3-pyrro!idinyl]- V-[ ,3]thiazolo[5,4-6]pyridin-2-y!-4-( ⁇ [(1 S)-1 ,2,2- trimethylpropyl]amino ⁇ methyl)-2,6-pyridinediamine difumarate monohydrate characterised in that it provides an XRPD pattern comprising peaks (°2 ⁇ ) at about 4.5, about 7.5 and/or about 8.7.
  • the present invention provides a polymorph of W-[(3S)-1-acryloyl- 3-pyrrolidinyl]-W-[1 ,3]thiazolo[5,4-i ) ]pyridm-2-yi-4-( ⁇ t ⁇ 1 S)-1 ,2,2- trimethylpropyl]amino ⁇ methyi)-2,6-pyridinediamine difumarate monohydrate characterised in that it provides an XRPD pattern comprising peaks substantially as set out in Table 1.
  • the present invention provides a polymorph of /V-[(3S)-1 -acryioyi- S-pyrrolidinylj-W-tL ⁇ thiazolors ⁇ -jbjpyhdin ⁇ -yl ⁇ - ⁇ I S)-! ⁇ - trimethy!propyi3amino ⁇ methyi)-2,6-pyridinediarriine difumarate monohydrate characterised in that it provides an XRPD pattern substantialiy in accordance with Figure 1.
  • the invention encompasses polymorphs isolated in pure form or when admixed with other materia!s, for example other polymorphs, or any other materia!.
  • a polymorph in isolated or pure form refers to a sample in which the polymorph is present in an amount of >75%, particularly >90%, more particularly >95% and even more particularly >99% relative to other materials which may be present in the sample.
  • the invention also includes isotopically-labelled compounds, which are identical to the compounds of formula (I) and salts thereof, but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number most commonly found in nature.
  • isotopes that can be incorporated into the compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen and fluorine, such as 2 H, 3 H, 11 C, 4 C and 18 F.
  • the compounds according to formuia (!) may contain one or more asymmetric center (also referred to as a chirai center) and may, therefore, exist as individual enantiomers, diastereomers, or other stereoisomer ⁇ forms, or as mixtures thereof.
  • Chirai centers such as chirai carbon atoms, may also be present in a substituent such as an aikyl group.
  • the stereochemistry of a chira! center present in formula (I), or in any chemical structure illustrated herein, is not specified the structure is intended to encompass any stereoisomer and all mixtures thereof.
  • compounds according to formula (I) containing one or more chirai center may be used as racemic mixtures, enantiomerically enriched mixtures, or as enantiomerically pure individual stereoisomers.
  • the invention provides W-II -acryloyl-S-p rrolidinyll-W-tl ⁇ lthiazoloIS ⁇ - b]pyridin-2-yi-4-( ⁇ [1 ,2,2-trimethylpropyl]amino ⁇ methy!)-2,6-pyridinediamine in all stereoisomeric forms, and mixtures thereof.
  • Individual stereoisomers of a compound according to formula (I) which contain one or more asymmetric center may be resolved by methods known to those skilled in the art. For example, such resolution may be carried out (1 ) by formation of diastereoisomeric salts, complexes or other derivatives; (2) by selective reaction with a stereoisomer- specific reagent, for example by enzymatic oxidation or reduction; or (3) by gas-iiquid or liquid chromatography in a chiral enviornment, for example, on a chiral support such as silica with a bound chiral ligand or in the presence of a chiral solvent.
  • stereoisomers may be synthesized by asymmetric synthesis using optically active reagents, substrates, catalysts or solvents, or by converting one enantiomer to the other by asymmetric transformation.
  • the compounds according to formula (I) may also contain centers of geometric asymmetry. Where the stereochemistry of a center of geometric asymmetry present in formula (I), or in any chemical structure illustrated herein, is not specified, the structure is intended to encompass the trans geometric isomer, the cis geometric isomer, and all mixtures thereof. Likewise, al! tautomeric forms are a!so included in formula (I) whether such tautomers exist in equilibrium or predominately in one form.
  • references herein to compounds of formula (I) and salts thereof covers the compounds of formula (I) as free acids or free bases, or as salts thereof, for example as pharmaceutically acceptable salts thereof.
  • the invention is directed to compounds of formula (!) as the free acid or free base.
  • the invention is directed to compounds of formula (I) and salts thereof.
  • the invention is directed to compounds of formula (I) and pharmaceutically acceptable salts thereof.
  • pharmaceuticaliy acceptable salts of the compounds according to formula (I) may be prepared. Indeed, in certain embodiments of the invention, pharmaceutically acceptable salts of the compounds according to formula (I) may be preferred over the respective free base or free acid because such salts impart greater stability or solubility to the molecule thereby facilitating formulation into a dosage form. Accordingly, the invention is further directed to compounds of formula (I) and pharmaceutically acceptable salts thereof.
  • pharmaceutically acceptable salts refers to salts that retain the desired biological activity of the subject compound and exhibit minimal undesired toxicological effects.
  • salts and solvates having non-pharmaceutically acceptable counter-ions or associated solvents are within the scope of the present invention, for example, for use as intermediates in the preparation of other compounds of formula (I) and their pharmaceutically acceptable salts.
  • one embodiment of the invention embraces compounds of formula (I) and salts thereof.
  • compounds according to formula (I) may contain an acidic functional group.
  • Suitable pharmaceuticaiiy-acceptable salts include salts of such acidic functional groups.
  • Representative salts include pharmaceutically acceptable metal salts such as sodium, potassium, lithium, calcium, magnesium, aluminum, and zinc salts; carbonates and bicarbonates of a pharmaceutically acceptable metal cation such as sodium, potassium, lithium, calcium, magnesium, aluminum, and zinc; pharmaceuticaily acceptable organic primary, secondary, and tertiary amines including aliphatic amines, aromatic amines, aliphatic diamines, and hydroxy alkylamines such as methylamine, ethylamine, 2-hydroxyethylamine, diethylamine, TEA, ethyienediamine, ethanolamine, diethanolamine, and cyclohexylamine.
  • compounds according to formula (I) may contain a basic functional group and are therefore capable of forming pharmaceutically acceptable acid addition salts by treatment with a suitable acid.
  • suitable acids include pharmaceutically acceptable inorganic acids and pharmaceutically acceptable organic acids.
  • Representative pharmaceutically acceptable acid addition salts include hydrochloride, hydrobromide, nitrate, methyinitrate, sulfate, bisulfate, sulfamate, phosphate, acetate, hydroxyacetate, phenylacetate, propionate, butyrate, isobutyrate, valerate, maleate, hydroxymaleate, acrylate, fumarate, maiate, tartrate, citrate, salicylate, p- aminosalicyclate, glycollate, lactate, heptanoate, phthalate, oxalate, succinate, benzoate, o-acetoxybenzoate, ch!orobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzo
  • the invention provides a compound which is A/-[(3S)-1-acryloyl-3- pyrrolEdiny]]-/V-[1 ,3]thiazolo[5,4-f)]pyridin-2-yl-4-( ⁇ [(1 S)-1 ,2,2- trimethy!propyl]amino ⁇ methyl)-2,6-pyridinediamine or a pharmaceutically acceptable salt thereof.
  • the invention provides a pharmaceutically acceptable salt of N- [(3S)-1 -acryjoyl-3-pyrrolidinyl3-/V-[1 ,3]thiazolo[5,4-/b3pyridin-2-yl-4-( ⁇ [(1 S)-1 ,2,2 ⁇ trimethylpropyl]amino ⁇ methyl)-2,6-pyridinediamine,
  • the invention provides salts of compounds of formula (!) formed by treatment with fumaric acid, in another embodiment, the invention provides difumarate salts of compounds of formula (I).
  • the invention provides ⁇ /- [ ⁇ 3S)-1-acryloyl-3-pyrrolidinyl]-/v 1 ⁇ [1 ,3]thiazolo[5,4-/b]pyridin-2-y!-4- ⁇ [(1 S)-1 ,2,2- trimethylpropy!]amino ⁇ methyl)-2,6-pyrtdinediamine difumarate.
  • the compounds of the invention may be made by a variety of methods, including standard chemistry. Any previously defined variable will continue to have the previously defined meaning unless otherwise indicated. Illustrative general synthetic methods are set out in the following schemes, and can be readily adapted to prepare other compounds of the invention. Specific compounds of the invention are prepared in the Examples section.
  • R 1 is -CH 2 OCH 3
  • R 2 is -NHR 29 NHR 30
  • R 3 is H
  • Y is N
  • R 29 is (CH 2 ) n or C 3 . 6 cycloalkyi
  • R 30 is -COCHCHR 21 , -COCH 2 halo or - COCH 2 OR 22
  • X is N and P is H or an amine protecting group such as Boc).
  • R 1 is H, 2-methyi-1 H-imidazoiyl or N- morpholinylCH 2
  • R 2 is -NHR 29 NHR 30
  • R 3 is H
  • Y is CH or N
  • R 29 is (CH 2 ) n or C 3 .
  • R 30 is -COCHCHR 21 , -COCH 2 halo or -COCH 2 OR 22
  • X is N and P is H or an amine protecting group such as Boc).
  • scheme 2 can be adapted to provide other compounds of formula (I), by replacing the ⁇ , ⁇ -diamine in the penultimate step with for instance (i) an a-hydroxy-to-(N-protected) amino compound (to give a compound of formula i wherein R 2 is OR 9 , or (ii) a phenyl boronic acid optionally substituted by -NR 10 R 11 , in the presence of a palladium catalyst, to give a compound of formula (I) wherein R 2 is phenyl optionally substituted by -NR 10 R 11 , or (iii) an ⁇ , ⁇ -diamine in which either one or both N are incorporated into a 6-membered heterocyclic ring.
  • Scheme 3
  • Compounds of formula (I) wherein R 3 is chloro may be reduced to produce compounds of formula (I) wherein R 3 is hydrogen.
  • Suitable reduction conditions include treatment with ammonium formate under microwave irradiation, in the presence of a suitable catalyst such as palladium on activated carbon, in a suitable solvent such as methanol and at a suitable temperature such as about 130 °C.
  • Compounds of formula (I) wherein R 2 , R 3 and X are as defined above and R 1 is - CH 2 NR 6 R 7 may be prepared from compounds of formula (I) wherein R 1 is -CH 2 OCH 3 by conversion of the methoxy group to the corresponding alcohol by treatment with, for example, boron tribromide in the presence of a suitable solvent such as dichloromethane and at a suitable temperature such as ambient temperature, followed by conversion of the alcohol to the corresponding aldehyde by treatment with, for example, manganese dioxide in the presence of a suitable solvent such as tetrahydrofuran and at a suitable temperature such as about 70°C, followed by reaction with an amine of formula R 6 R 7 NH 2 in the presence of sodium triacetoxyborohydride, a suitable solvent such as dichloromethane and at a suitable temperature such as ambient temperature.
  • Compounds of formula (I) wherein R 2 , R 3 and X are as defined above and R 1 is -CH 2 NH 2 may be prepared from the abovementioned aldehyde by conversion to the corresponding oxime by treatment with, for example, hydroxylamine hydrochloride and sodium acetate in the presence of a suitable sovent such as aqueous ethanol and at a suitable temperature such as ambient temperature, followed by treatment with zinc powder in glacial acetic acid at a suitable temperature such as ambient temperature.
  • a suitable sovent such as aqueous ethanol
  • a suitable temperature such as ambient temperature
  • the invention provides a process for preparing a compound of formula (I) comprising:
  • R 1 , R 3 , X and Y are as defined above and R 2 is NR 7 R 8 wherein R 7 is as defined above and R 8 is -(CH 2 ) n NR 12 R 13 , C 3 . 6 cycloalkyl substituted by -NR 4 R 15 , or 5- or 6- membered heterocyclyl wherein the 5- or 6-membered heterocyclyl contains a nitrogen atom and is substituted by -R 6 , or R 7 and R 8 , together with the nitrogen atom to which they are attached, are linked to form a 6-membered heterocyciyi optionally containing a further nitrogen atom wherein the 6- membered heterocyciyi is substituted by -NR 18 R 19 ;
  • R ⁇ R 3 , X and Y are as defined above and
  • Y 1 is:
  • R 88 is 5- or 6-membered heterocyciyi which contains a nitrogen atom, a 6-membered heterocyciyi containing, and linked through nitrogen, and optionally containing a further nitrogen atom, wherein the 6-membered heterocyciyi is substituted by -NHR 18 ;
  • R 99 is 5- or 6-membered heterocyciyi wherein the 5- or 6-membered heterocyciyi contains a nitrogen atom, or
  • R a is R 11 , R 13 , R 15 , R 16 , R 19 or R 20 are as defined above;
  • a suitable solvent such as /V-methy!-2-pyrrolidone
  • a base such as a tertiary amine such ty/V-diisopropyiethy!amine
  • a suitable temperature such as ambient temperature, or
  • R a is R 11 , R 3 , R 15 , R 16 , R 19 or R 20 are as defined above,
  • the compounds of the invention are inhibitors of kinase activity, in particular Itk activity.
  • Compounds which are Itk inhibitors may be useful in the treatment of disorders wherein the underlying pathology is (at least in part) attributable to inappropriate Itk activity, such as asthma.
  • "Inappropriate Itk activity” refers to any Itk activity that deviates from the normal Itk activity expected in a particular patient. Inappropriate Itk may take the form of, for instance, an abnormal increase in activity, or an aberration in the timing and or control of Itk activity. Such inappropriate activity may result then, for example, from overexpression or mutation of the protein kinase leading to inappropriate or uncontrolled activation. Accordingly, in another aspect the invention is directed to methods of treating such disorders.
  • Such disorders include respiratory diseases including asthma, chronic obstructive pulmonary disease (COPD) and bronchitis; allergic diseases including allergic rhinitis and atopic dermatitis; autoimmune diseases including rheumatoid arthritis, multiple sclerosis, psoriasis, type I diabetes, T cell mediated hypersensitivities, Guillain-Barre Syndrome and Hashimoto's thyroiditis; transplant rejection; graft versus host disease; inflammatory disorders including conjunctivitis, contact dermatitis, inflammatory bowel disease and chronic inflammation; HIV; aplastic anemia; and pain including inflammatory pain.
  • respiratory diseases including asthma, chronic obstructive pulmonary disease (COPD) and bronchitis
  • allergic diseases including allergic rhinitis and atopic dermatitis
  • autoimmune diseases including rheumatoid arthritis, multiple sclerosis, psoriasis, type I diabetes, T cell mediated hypersensitivities, Guillain-
  • the disorder mediated by inappropriate Itk activity is a respiratory disease; an allergic disease; an autoimmune disease; transplant rejection; graft versus host disease; an inflammatory disorder; HIV; aplastic anemia; or pain.
  • the disorder mediated by inappropriate Itk activity is asthma, chronic obstructive pulmonary disease (COPD), bronchitis, allergic rhinitis, atopic dermatitis, rheumatoid arthritis, multiple sclerosis, psoriasis, type i diabetes, T cell mediated hypersensitivity, Guillain-Barre Syndrome, Hashimoto's thyroiditis, transplant rejection, graft versus host disease, conjunctivitis, contact dermatitis, inflammatory bowel disease, chronic inflammation, HIV, aplastic anemia, or inflammatory pain.
  • COPD chronic obstructive pulmonary disease
  • the disorder mediated by inappropriate Itk activity is asthma.
  • the methods of treatment of the invention comprise administering a safe and effective amount of a compound of formula (I) or a pharmaceuticaliy acceptable sa!t thereof to a patient in need thereof.
  • Individual embodiments of the invention include methods of treating any one of the above-mentioned disorders by administering a safe and effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof to a patient in need thereof.
  • treat in reference to a disorder means: (1 ) to ameliorate or prevent the disorder or one or more of the biological manifestations of the disorder, (2) to interfere with (a) one or more points in the biological cascade that leads to or is responsible for the disorder or (b) one or more of the biological manifestations of the disorder, (3) to alleviate one or more of the symptoms or effects associated with the disorder, or (4) to slow the progression of the disorder or one or more of the biological manifestations of the disorder.
  • prevention of a disorder includes prevention of the disorder.
  • prevention is not an absolute term. In medicine, “prevention” is understood to refer to the prophylactic administration of a drug to substantially diminish the likelihood or severity of a disorder or biological manifestation thereof, or to delay the onset of such disorder or biological manifestation thereof.
  • safe and effective amount in reference to a compound of formula (I) or a pharmaceutically acceptable salt thereof or other pharmaceutica!ly-active agent means an amount of the compound sufficient to treat the patient's condition but low enough to avoid serious side effects (at a reasonable benefit/risk ratio) within the scope of sound medical judgment.
  • a safe and effective amount of a compound will vary with the particular compound chosen (e.g.
  • patient refers to a human (including adults and children) or other animal, in one embodiment, “patient” refers to a human.
  • the compounds of formula (I) or pharmaceutically acceptable salts thereof may be administered by any suitable route of administration, including both systemic administration and topical administration.
  • Systemic administration includes oral administration, parenteral administration, transdermal administration and recta! administration.
  • Parenteral administration refers to routes of administration other than enteral or transdermal, and is typically by injection or infusion.
  • Parenteral administration includes intravenous, intramuscular, and subcutaneous injection or infusion.
  • Topical administration includes application to the skin as well as intraocular, otic, intravaginal, inhaled and intranasal administration.
  • Inhalation refers to administration into the patient's lungs whether inhaled through the mouth or through the nasal passages.
  • the compounds of formula (I) or pharmaceutically acceptable salts thereof may be administered orally.
  • the compounds of formula (I) or pharmaceutically acceptable salts thereof may be administered topically.
  • the compounds of formula (! or pharmaceutically acceptable salts thereof may be administered by inhalation.
  • the compounds of formula (I) or pharmaceutically acceptable salts thereof may be administered intranasally.
  • the compounds of formula (I) or pharmaceutically acceptable salts thereof may be administered once or according to a dosing regimen wherein a number of doses are administered at varying intervals of time for a given period of time. For example, doses may be administered one, two, three, or four times per day. In one embodiment, a dose is administered once per day. in a further embodiment, a dose is administered twice per day. Doses may be administered until the desired therapeutic effect is achieved or indefinitely to maintain the desired therapeutic effect. Suitable dosing regimens for a compound of formula (I) or a pharmaceutically acceptable salt thereof depend on the pharmacokinetic properties of that compound, such as absorption, distribution, and half- life, which can be determined by the skilled artisan.
  • suitable dosing regimens including the duration such regimens are administered, for a compound of formula (I) or a pharmaceutically acceptable salt thereof depend on the disorder being treated, the severity of the disorder being treated, the age and physical condition of the patient being treated, the medical history of the patient to be treated, the nature of concurrent therapy, the desired therapeutic effect, and like factors within the knowledge and expertise of the skilled artisan. It will be further understood by such skilled artisans that suitable dosing regimens may require adjustment given an individual patient's response to the dosing regimen or over time as individual patient needs change. Typical daily dosages may vary depending upon the particular route of administration chosen.
  • Typical daily dosages for oral administration range from 0.001 mg to 50mg per kg of total body weight, for example from 1 mg to 10mg per kg of total body weight.
  • daily dosages for oral administration may be from Q.5mg to 2g per patient, such as 10mg to 1g per patient.
  • a prodrug of a compound of formula (I) is a functional derivative of the compound which, upon administration to a patient, eventually liberates the compound of formula (I) in vivo.
  • Administration of a compound of formula (I) as a prodrug may enable the skilled artisan to do one or more of the following: (a) modify the onset of the activity of the compound in vivo; (b) modify the duration of action of the compound in vivo; (c) modify the transportation or distribution of the compound in vivo; (d) modify the solubility of the compound in vivo; and (e) overcome a side effect or other difficulty encountered with the compound.
  • Typical functional derivatives used to prepare prodrugs include modifications of the compound that are chemically or enzymaticaliy cleavable in vivo. Such modifications, which include the preparation of phosphates, amides, esters, thioesters, carbonates, and carbamates, are well known to those skilled in the art.
  • the invention thus provides a method of treating a disorder mediated by inappropriate itk activity comprising administering a safe and effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof to a patient in need thereof.
  • the invention provides a method of treating a disorder mediated by inappropriate Itk activity comprising administering a safe and effective amount of N-[(ZS)- 1 -acryloyl-S-pyrro!idinylj-W-tl ,33thiazolo[5,4-0]pyridin-2-yi-4-( ⁇ [(1 S)-1 ,2,2- trimethylpropyi]amino ⁇ methyl)-2,6-pyridinediamine or a pharmaceuticaily acceptable salt thereof to a patient in need thereof.
  • the invention provides a method of treating a disorder mediated by inappropriate Itk activity comprising administering a safe and effective amount of /V-[(3S)-1-acryloyl-3-pyrrolidinyl3-W- [1 ,3]thiazolo[5,4-6]pyridin-2-yl-4-( ⁇ [ ⁇ 1 S)- ,2,2-trimethylpropyl]amino ⁇ methyl)-2,6- pyridinediamine to a patient in need thereof.
  • the invention provides a method of treating a disorder mediated by inappropriate Itk activity comprising administering a safe and effective amount of A/-[(3S)-1-acryloyl-3 ⁇ pyrrolidinyl3-/V- [1 ,3jthiazolo[5,4-0]pyridin-2-yl-4-( ⁇ [(1 S)-1 ,2,2-trimethy!propyl]amino ⁇ methyl)-2,6- pyridinediamine difumarate to a patient in need thereof.
  • the disorder mediated by inappropriate Itk activity is selected from the group consisting of respiratory diseases (including asthma, chronic obstructive pulmonary disease (COPD) and bronchitis); allergic diseases (including allergic rhinitis and atopic dermatitis); autoimmune diseases (including rheumatoid arthritis, multiple sclerosis, psoriasis, type I diabetes, T cell mediated hypersensitivities, Guillain-Barre Syndrome and Hashimoto's thyroiditis); transplant rejection; graft versus host disease; inflammatory disorders (including conjunctivitis, contact dermatitis, inflammatory bowel disease and chronic inflammation); HIV; aplastic anemia; and pain including inflammatory pain.
  • respiratory diseases including asthma, chronic obstructive pulmonary disease (COPD) and bronchitis
  • allergic diseases including allergic rhinitis and atopic dermatitis
  • autoimmune diseases including rheumatoid arthritis, multiple sclerosis, psoriasis, type I diabetes
  • the disorder mediated by inappropriate Itk activity is a respiratory disease.
  • the disorder mediated by inappropriate Itk activity is asthma.
  • the invention provides a method of treating a respiratory disease comprising administering a safe and effective amount of /V-[(3S)-1-acryloyl-3-pyrrolidinylj- W-[1 ,3]thiazolo[5,4-0]pyridin-2-yl-4-( ⁇ [(1 S)-1 ,2,2-trimethylpropyl]amino ⁇ methyl)-2,6- pyridinediamine or a pharmaceutically acceptable salt thereof to a patient in need thereof.
  • the invention provides a method of treating a respiratory disease comprising administering a safe and effective amount of A/-[(3S)-1-acryloy!-3-pyrrolidinyl]- ,3]thiazolo[5,4-/f)jpyridin-2-y!-4-( ⁇ [(1 S)-1 ,2,2-trimethylpropyl]amino ⁇ methyl)-2,6- pyridinediamine to a patient in need thereof.
  • the invention provides a method of treating a respiratory disease comprising administering a safe and effective amount of W-[(3S)-1 -acryloy!-3-pyrrolidinyl]-W-[1 ,3]thiazolo[5,4-b]pyridin-2-yl-4- ( ⁇ [(1 S)-1 ,2,2-trimethylpropyl]amino ⁇ methyl)-2,6-pyridtnediamine difumarate to a patient in need thereof.
  • the invention provides a method of treating asthma comprising administering a safe and effective amount of W-[(3S)-1-acry!oyl-3-pyrroiidinyl]-/v ⁇ [1 ,3]thiazolo[5,4-/bjpyridin-2-yl-4-( ⁇ [(1 S)-1 ,2,2-trimethyipropyl]amino ⁇ methyl)-2,6- pyridinediamine or a pharmaceutically acceptable salt thereof to a patient in need thereof.
  • the invention provides a method of treating asthma comprising administering a safe and effective amount of W-[ ⁇ 3S)-1-acryloyl-3-pyrrolidinyl]-/V- [1 ,3]thiazo!o[5,4-0]pyridin-2-yl-4-( ⁇ [(1 S)-1 ,2,2-trimethylpropy!]amino ⁇ methyi)-2,6- pyridinediamine to a patient in need thereof.
  • the invention provides a method of treating asthma comprising administering a safe and effective amount of W-[(3S)-1 -acryloyl-3-pyrrolidinyl]-A "-[1 ,3]thiazolo[5,4- >]pyridin-2-yl ⁇ 4-( ⁇ [(1 S)- l ⁇ -tnmethylpropyljaminoJmethy ⁇ -pyridinediarnine difumarate to a patient in need thereof.
  • the invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in medical therapy.
  • the invention provides V-f(3S)-1 -acryloyl-3-pyrrolidinyl)-/V-[1 ,3]thia2oSot5,4- 6]pyridin-2-y!-4-( ⁇ [(1 S)-1 ,2,2-trimethylpropyl]amino ⁇ methyl)-2,6-pyridinediamine or a pharmaceutically acceptable salt thereof for use in medical therapy
  • the invention provides A/-[(3S)-1-acryloyl-3-pyrrolidinyl]-A/ , -[1 ,3]thiazolo[5,4- 0]pyridin-2-yl-4- ⁇ [(1 S)-1 ,2,2-trimethylpropyi]amino ⁇ methyl)-2,6-pyhdinediamine for use in medical therapy, in a further embodiment, the invention provides A/-[[(3S)-1 -acryloyl
  • the invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in the treatment of a disorder mediated by inappropriate Itk activity.
  • the invention provides W-[(3S)-1- acryloy!-3-pyrroiidinyl]-W-[1 ,3]th!azolo[5,4-0]pyridin-2-y!-4-( ⁇ [ ⁇ 1 S)-1 ,2,2- trimethylpropyl]amino ⁇ methyl)-2,6-pyridinediamine or a pharmaceutically acceptable salt thereof for use in the treatment of a disorder mediated by inappropriate Itk activity.
  • the invention provides W-[(3S)-1-acryloyi-3-pyrrolidiny!]-A/'- [1 ,3]thiazo!o[5,4-jb]pyridin-2-yl-4-( ⁇ [ ⁇ 1 S)-1 ,2,2-trimethylpropyS]amino ⁇ methy!-2,6- pyridinediamine for use in the treatment of a disorder mediated by inappropriate Itk activity.
  • the invention provides A/-[(3S)-1-acryloyl-3-pyrrolidinyl]- ,33thiazolo[5,4- )]pyridin-2-yl-4-( ⁇ [ ⁇ 1 S)-1 ,2,2-trimethy!propyl]amino ⁇ methyl)-2,6- pyridinediamine difumarate for use in the treatment of a disorder mediated by inappropriate Itk activity.
  • the invention provides the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for use in the treatment of a disorder mediated by inappropriate Itk activity.
  • the invention provides the use of A/-[(3S)-1-acryloyl-3-pyrrolidinyl3-/V-[1 ,3]thiazolo[5 > 4- jt>jpyridin-2-yl-4-( ⁇ [(1 S)-1 ,2,2-tnmethylpropyl]amino ⁇ methyl)-2,6-pyhdinediamine or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for use in the treatment of a disorder mediated by inappropriate itk activity.
  • the invention provides the use of A/-[(3S)-1 -acryloyl-3-pyrrolidinyl]-A '-[1 ,3]thiazolo[5,4- jb]pyridin-2-yl-4-( ⁇ [(1 S)-1 ⁇ -tnmethylpropyljaminojmethyl ⁇ .e-pyridinediamine in the manufacture of a medicament for use in the treatment of a disorder mediated by inappropriate Itk activity, in a further embodiment, the invention provides the use of N- [(3S)-1-acry!oyl-3-pyrrolidinyl]-A/ , -[1 ,3]thiazolo[5,4-j ]pyridin-2-yl-4-( ⁇ [(1 S)-1 ,2,2- trimethyipropy!]amino ⁇ methyl)-2,6-pyridinediamine difumarate in the manufacture of a medicament for use in the treatment of a disorder mediated by inappropriate Itk
  • the invention provides a pharmaceutical composition for the treatment or prophylaxis of a disorder mediated by inappropriate Itk activity comprising a compound of formula (I) or a pharmaceuticaliy acceptable salt thereof.
  • the invention provides a pharmaceutical composition for the treatment or prophylaxis of a disorder mediated by inappropriate Itk activity comprising W-[(3S)-1- acryloyl-3-pyrroiidinyl]-/V-[1 ,3]thiazolo[5,4-0]pyridin-2-y!-4-( ⁇ [(1 S)-1 ,2,2- trimethylpropyl]amino ⁇ methyi)-2,6-pyridinediamine or a pharmaceutically acceptable salt thereof.
  • the invention provides a pharmaceutical composition for the treatment or prophylaxis of a disorder mediated by inappropriate Itk activity comprising W-[(3S)-1-acryloyl-3-pyrrolidinyi]-W-[1 ,3]thtazolo[5,4-i)]pyridin-2-yi-4- ⁇ [(1 S)- ,2,2- trimethy!propyl]amino ⁇ methyl)-2,6-pyridinediamine.
  • the invention provides a pharmaceutical composition for the treatment or prophylaxis of a disorder mediated by inappropriate Itk activity comprising W-[(3S)-1 -acryloyl-3-pyrrolidinyl]- v 1 - [1 ,3]thiazolo[5,4-0]pyridin-2-yl-4-( ⁇ [(1 S)-1 ,2,2-trimethylpropyl]amino ⁇ methyl)-2,6- pyridinediamine difumarate.
  • the compounds of formula (I) and pharmaceutically acceptable salts thereof will normally, but not necessarily, be formulated into pharmaceutical compositions prior to administration to a patient.
  • the invention is directed to pharmaceutical compositions comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable excipients.
  • the invention provides a pharmaceutical composition
  • a pharmaceutical composition comprising N- [(SSH-acryioy!-S-pyrrolidiny!l-W-EI ,3]thiazolo[5,4-6]pyridin-2-yl-4-( ⁇ [(1 S)-1 ,2,2- thmethylpropyi3amino ⁇ methyl)-2,6-pyridinediamine or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable excipients.
  • the invention provides a pharmaceutical composition
  • a pharmaceutical composition comprising W-[(3S)-1-acryloyl-3-pyrrolidinyl]-W-[1 ,33thiazolo[5,4-/3]pyridin-2-yl-4-( ⁇ [(1 S)-1 ,2,2- trimethylpropyl]amino ⁇ methyi)-2,6-pyridinediamine and one or more pharmaceutically acceptable excipients.
  • the invention provides a pharmaceutical composition
  • a pharmaceutical composition comprising W-[ ⁇ 3S)-1-acryloyl-3-pyrrolidinyl]-/V-[1 ,3]thiazolo[5,4-6]pyridin-2-yl-4-( ⁇ [(1 S)-1 ,2,2- trimethylpropyl]amino ⁇ methyl)-2,6-pyridinediamine difumarate and one or more pharmaceutically acceptable excipients.
  • the invention is directed to pharmaceutical compositions for the treatment or prophylaxis of a disorder mediated by inappropriate Itk activity comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof.
  • the pharmaceutical compositions of the invention may be prepared and packaged in bulk form wherein a safe and effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof can be extracted and then given to the patient such as with powders or syrups.
  • the pharmaceutical compositions of the invention may be prepared and packaged in unit dosage form wherein each physically discrete unit contains a compound of formula (I) or a pharmaceutically acceptable salt thereof.
  • the pharmaceutical compositions of the invention typically may contain, for example, from 0.5mg to 1 g, or from 1 mg to 700mg, or from 5mg to 100mg of a compound of formula (I) or a pharmaceutically acceptable salt thereof.
  • compositions of the invention typically contain one compound of formula (I) or a pharmaceutically acceptable salt thereof.
  • pharmaceutically acceptable excipient means a pharmaceutically acceptable material, composition or vehicle involved in giving form or consistency to the pharmaceutical composition.
  • Each excipient must be compatible with the other ingredients of the pharmaceutical composition when commingled such that interactions which would substantially reduce the efficacy of the compound of formula (I) or a pharmaceutically acceptable salt thereof when administered to a patient and interactions which would result in pharmaceutical compositions that are not pharmaceutically acceptable are avoided.
  • each excipient must of course be pharmaceutically acceptable eg of sufficiently high purity.
  • dosage forms include those adapted for (1 ) oral administration such as tablets, capsules, caplets, piils, troches, powders, syrups, elixers, suspensions, solutions, emulsions, sachets, and cachets; (2) parenteral administration such as sterile solutions, suspensions, and powders for reconstitution; (3) transdermal administration such as transdermal patches; (4) rectal administration such as suppositories; (5) inhalation such as aerosols, solutions, and dry powders; and (6) topical administration such as creams, ointments, lotions, solutions, pastes, sprays, foams, and gels.
  • Suitable pharmaceutically acceptable excipients will vary depending upon the particular dosage form chosen.
  • suitable pharmaceutically acceptable excipients may be chosen for a particular function that they may serve in the composition.
  • certain pharmaceutically acceptable excipients may be chosen for their ability to facilitate the production of uniform dosage forms.
  • Certain pharmaceutically acceptable excipients may be chosen for their ability to facilitate the production of stable dosage forms.
  • Certain pharmaceutically acceptable excipients may be chosen for their ability to facilitate the carrying or transporting of the compound or compounds of formula (I) or pharmaceutically acceptable salts thereof once administered to the patient from one organ, or portion of the body, to another organ, or portion of the body.
  • Certain pharmaceutically acceptable excipients may be chosen for their ability to enhance patient compliance.
  • Suitable pharmaceuticaliy-acceptabie excipients include the following types of excipients: Diluents, fillers, binders, disintegrants, lubricants, glidants, granulating agents, coating agents, wetting agents, solvents, co-solvents, suspending agents, emulsifiers, sweetners, flavoring agents, flavor masking agents, coloring agents, anticaking agents, hemectants, chelating agents, plasticizers, viscosity increasing agents, antioxidants, preservatives, stabilizers, surfactants, and buffering agents.
  • compositions of the invention are prepared using techniques and methods known to those skilled in the art. Some of the methods commonly used in the art are described in Remington's Pharmaceutical Sciences (Mack Publishing Company). Accordingly, in another aspect the invention is directed to process for the preparation of a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof and one or more pharmaceutically-acceptable excipients which comprises mixing the ingredients.
  • a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof may be prepared by, for example, admixture at ambient temperature and atmospheric pressure.
  • the compounds of formula (! or pharmaceutically acceptable salts thereof will be formulated for oral administration.
  • the compounds of formula (I) or pharmaceutically acceptable salts thereof will be formulated for inhaled administration.
  • the compounds of formula (I) or pharmaceutically acceptable salts thereof will be formulated for intranasal administration.
  • the invention is directed to a solid oral dosage form such as a tablet or capsule comprising a safe and effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof and a diluent or filler.
  • Suitable diluents and fillers include lactose, sucrose, dextrose, mannitol, sorbitol, starch (e.g. corn starch, potato starch, and pre-gelatinized starch), cellulose and its derivatives (e.g. microcrystalline cellulose), calcium sulfate, and dibasic calcium phosphate.
  • the oral solid dosage form may further comprise a binder. Suitable binders include starch (e.g.
  • the oral solid dosage form may further comprise a disintegrant. Suitable disintegrants include crospovidone, sodium starch glycolate, croscarmelose, alginic acid, and sodium carboxymethyl cellulose.
  • the oral solid dosage form may further comprise a lubricant. Suitable lubricants include stearic acid, magnesuim stearate, calcium stearate, and talc.
  • dosage unit formulations for oral administration can be microencapsulated.
  • the composition can also be prepared to prolong or sustain the release as for example by coating or embedding particulate material in polymers, wax or the like.
  • the compounds of formula (I) or pharmaceutically acceptable salts thereof may also be coupled with soluble polymers as targetabie drug carriers.
  • soluble polymers can include polyvinylpyrrolidone, pyran copolymer, polyhydroxypropylmethacrylamide -phenol, po!yhydroxyethy!aspartamidephenol, or poiyethyleneoxidepolylysine substituted with palmitoyl residues.
  • the compounds of formula (I) or pharmaceutically acceptable salts thereof may be coupled to a class of biodegradable polymers useful in achieving controlled release of a drug, for example, polylactic acid, polepsilon caproiactone, polyhydroxy butyric acid, polyorthoesters, polyacetals, polydihydropyrans, poiycyanoacry!ates and cross-linked or amphipathic block copolymers of hydrogels.
  • a class of biodegradable polymers useful in achieving controlled release of a drug, for example, polylactic acid, polepsilon caproiactone, polyhydroxy butyric acid, polyorthoesters, polyacetals, polydihydropyrans, poiycyanoacry!ates and cross-linked or amphipathic block copolymers of hydrogels.
  • the invention is directed to a liquid oral dosage form.
  • Oral liquids such as solution, syrups and elixirs can be prepared in dosage unit form so that a given quantity contains a predetermined amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof.
  • Syrups can be prepared by dissolving the compound of formula (I) or a pharmaceutically acceptable salt thereof in a suitably flavored aqueous solution, while elixirs are prepared through the use of a non-toxic alcoholic vehicle.
  • Suspensions can be formulated by dispersing the compound of formula (I) or a pharmaceutically acceptable salt thereof in a non-toxic vehicle.
  • So!ubilizers and emulsifiers such as ethoxyiated isostearyl alcohols and polyoxy ethylene sorbitol ethers, preservatives, flavor additive such as peppermint oil or natural sweeteners or saccharin or other artificial sweeteners, and the like can also be added.
  • the invention is directed to a dosage form adapted for administration to a patient by inhalation.
  • a dosage form adapted for administration to a patient by inhalation.
  • dry powder compositions for delivery to the lung by inhalation typically comprise a compound of formula (i) or a pharmaceutically acceptable salt thereof as a finely divided powder together with one or more pharmaceuticaily-acceptable excipients as finely divided powders.
  • Pharmaceuticaily-acceptable excipients particularly suited for use in dry powders are known to those skilled in the art and include lactose, starch, mannitol, and mono-, di-, and polysaccharides.
  • the finely divided powder may be prepared by, for example, micronisation and milling.
  • the size-reduced (eg micronised) compound can be defined by a D 50 value of about 1 to about 10 microns (for example as measured using laser diffraction).
  • the dry powder may be administered to the patient via a reservoir dry powder inhaler (RDPI) having a reservoir suitable for storing multiple (un-metered doses) of medicament in dry powder form.
  • RDPIs typically include a means for metering each medicament dose from the reservoir to a delivery position.
  • the metering means may comprise a metering cup, which is movable from a first position where the cup may be filled with medicament from the reservoir to a second position where the metered medicament dose is made available to the patient for inhalation.
  • the dry powder may be presented in capsules (e.g. gelatin or plastic), cartridges, or blister packs for use in a multi-dose dry powder inhaler (MDPl).
  • MDPl multi-dose dry powder inhaler
  • MDPIs are inhalers wherein the medicament is comprised within a multi-dose pack containing (or otherwise carrying) multiple defined doses (or parts thereof) of medicament.
  • the dry powder When the dry powder is presented as a blister pack, it comprises multiple blisters for containment of the medicament in dry powder form.
  • the blisters are typically arranged in regular fashion for ease of release of the medicament therefrom.
  • the blisters may be arranged in a generally circular fashion on a disc-form blister pack, or the blisters may be elongate in form, for example comprising a strip or a tape.
  • Each capsule, cartridge, or blister may, for example, contain between 20 ⁇ g-10mg of the compound of formula (I) or a pharmaceutically acceptable salt thereof.
  • Aerosols may be formed by suspending or dissolving a compound of formula (I) or a pharmaceutically acceptable salt thereof in a liquified propellant.
  • Suitable propellants include halocarbons, hydrocarbons, and other liquified gases.
  • propellants include: trichlorofluoromethane (propellant 1 1), dichlorofluoromethane (propellant 12), dichlorotetrafluoroethane (propellant 114), tetraf!uoroethane (HFA-134a), 1 ,1 - dif!uoroethane (HFA-152a), difluoromethane (HFA-32), pentafluoroethane (HFA-12), heptaf!uoropropane (HFA-227a), perfluoropropane, perfluorobutane, perfluoropentane, butane, isobutane, and pentane. Aerosols comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof will typically be administered to a patient via a metered dose inhaler (MD!). Such devices are known to those skilled in the art.
  • MD metered dose inhaler
  • the aerosol may contain additional pharmaceutical!y-acceptabie excipients typically used with MDIs such as surfactants, lubricants, cosclerosiss and other excipients to improve the physical stability of the formulation, to improve valve performance, to improve solubility, or to improve taste.
  • additional pharmaceutical!y-acceptabie excipients typically used with MDIs such as surfactants, lubricants, cosclerosiss and other excipients to improve the physical stability of the formulation, to improve valve performance, to improve solubility, or to improve taste.
  • a pharmaceutical aerosol formulation comprising a compound of formula (i) or a pharmaceutically acceptable salt thereof and a fluorocarbon or hydrogen-containing ch!orofluorocarbon as propellant, optionally in combination with a surfactant and/or a cosolvent,
  • a pharmaceutical aerosol formulation wherein the propellant is selected from 1 ,1 ,1 ,2-tetrafluoroethane, 1 ,1 , 1 ,2,3,3,3-heptafluoro-n-propane and mixtures thereof.
  • compositions of the invention may be buffered by the addition of suitable buffering agents.
  • Capsules and cartridges for use in an inhaler or insufflator may be formulated containing a powder mix for inhalation of a compound of formula (I) or a pharmaceutically acceptable salt thereof and a suitable powder base such as lactose or starch.
  • a powder mix for inhalation of a compound of formula (I) or a pharmaceutically acceptable salt thereof and a suitable powder base such as lactose or starch.
  • Each capsule or cartridge may generally contain from 20 ⁇ 9 to 10mg of the compound of formula (I) or pharmaceutically acceptable salt thereof.
  • the compound of formula (I) or pharmaceutically acceptable salt thereof may be presented without excipients such as lactose.
  • the proportion of the active compound of formula (I) or pharmaceutically acceptable salt thereof in the local compositions according to the invention depends on the precise type of formulation to be prepared but will generally be within the range of from 0.001 to 10% by weight. Generally, for most types of preparations, the proportion used will be within the range of from 0.005 to 1 %, for example from 0.01 to 0.5%. However, in powders for inhalation or insufflation the proportion used will normally be within the range of from 0.1 to 5%.
  • Aerosol formulations are preferably arranged so that each metered dose or "puff of aerosol contains from 20 g to 10mg, preferably from 20 ⁇ g to 2000 ⁇ g > more preferably from about 20 ⁇ 9 to 500 ⁇ g of a compound of formula (I).
  • Administration may be once daily or several times daily, for example 2, 3, 4 or 8 times, giving for example 1 , 2 or 3 doses each time.
  • the overall daily dose with an aerosol will be within the range from 100 ⁇ g to 10mg, preferably from 200 ⁇ g to 2000 ⁇ g.
  • the overall daily dose and the metered dose delivered by capsules and cartridges in an inhaler or insufflator will generally be double that delivered with aerosol formulations.
  • the particle size of the particulate (e.g., mtcronised) drug should be such as to permit inhalation of substantially all the drug into the lungs upon administration of the aerosol formulation and will thus be less than 100 microns, desirably less than 20 microns, and in particular in the range of from 1 to 10 microns, such as from 1 to 5 microns, more preferably from 2 to 3 microns.
  • the formulations of the invention may be prepared by dispersal or dissolution of the medicament and a compound of formula (I) or a pharmaceutically acceptable salt thereof in the selected prope!lant in an appropriate container, for example, with the aid of sonication or a high-shear mixer.
  • the process is desirably carried out under controlled humidity conditions.
  • the chemical and physical stability and the pharmaceutical acceptability of the aerosol formulations according to the invention may be determined by techniques well known to those skilled in the art.
  • the chemical stability of the components may be determined by HPLC assay, for example, after prolonged storage of the product.
  • Physical stability data may be gained from other conventional analytical techniques such as, for example, by leak testing, by valve delivery assay (average shot weights per actuation), by dose reproducibility assay (active ingredient per actuation) and spray distribution analysis.
  • the stability of the suspension aerosol formulations according to the invention may be measured by conventional techniques, for example, by measuring flocculation size distribution using a back light scattering instrument or by measuring particle size distribution by cascade impaction or by the "twin impinger” analytical process.
  • twin impinger assay means "Determination of the deposition of the emitted dose in pressurised inhalations using apparatus A” as defined in British Pharmacopaeia 1988, pages A204-207, Appendix XVII C.
  • Such techniques enable the "respirable fraction" of the aerosol formulations to be calculated.
  • MDI means a unit comprising a can, a secured cap covering the can and a formulation metering valve situated in the cap.
  • MDI system includes a suitable channelling device. Suitable channelling devices comprise for example, a valve actuator and a cylindrical or cone-like passage through which medicament may be delivered from the filled canister via the metering valve to the nose or mouth of a patient such as a mouthpiece actuator.
  • MDI canisters generally comprise a container capable of withstanding the vapour pressure of the propellant used such as a plastic or plastic-coated glass bottle or preferably a metal can, for example, aluminium or an alloy thereof which may optionally be anodised, lacquer-coated and/or plastic-coated (for example incorporated herein by reference WO96/32099 wherein part or all of the internal surfaces are coated with one or more fluorocarbon polymers optionally in combination with one or more non-fluorocarbon polymers), which container is closed with a metering valve.
  • the cap may be secured onto the can via ultrasonic welding, screw fitting or crimping.
  • MDIs taught herein may be prepared by methods of the art (e.g.
  • the canister is fitted with a cap assembly, wherein a drug-metering valve is situated in the cap, and said cap is crimped in place.
  • the metallic internal surface of the can is coated with a fluoropolymer, more preferably blended with a non-fluoropolymer.
  • the metailic internal surface of the can is coated with a polymer blend of polytetrafluoroethyiene (PTFE) and polyethersulfone (PES).
  • the whole of the metallic internal surface of the can is coated with a polymer blend of polytetrafluoroethyiene (PTFE) and polyethersulfone (PES).
  • PTFE polytetrafluoroethyiene
  • PES polyethersulfone
  • the metering valves are designed to deliver a metered amount of the formu!ation per actuation and incorporate a gasket to prevent leakage of propellant through the valve.
  • the gasket may comprise any suitable elastomeric materia!
  • valves are commercially available from manufacturers well known in the aerosol industry, for example, from Valois, France (e.g. DF10, DF30, DF60), Bespak pic, UK (e.g. BK300, BK357) and 3M-
  • the DIs may also be used in conjunction with other structures such as, without limitation, overwrap packages for storing and containing the MD!s, including those described in U.S. Patent Nos. 6,1 19,853; 6, 179,1 18; 6,315,112; 6,352,152; 6,390,291 ; and 6,679,374, as well as dose counter units such as, but not limited to, those described in U.S. Patent Nos. 6,360,739 and 6,431 ,168.
  • a metering valve is crimped onto an aluminium can to form an empty canister.
  • the particulate medicament is added to a charge vessel and liquefied propellant together with the optional excipients is pressure filled through the charge vessel into a manufacturing vessel.
  • the drug suspension is mixed before recirculation to a filling machine and an aliquot of the drug suspension is then filled through the metering valve into the canister.
  • a metering valve is crimped onto an aluminium can to form an empty canister.
  • the liquefied propellant together with the optional excipients and the dissolved medicament is pressure filled through the charge vessel into a manufacturing vessel.
  • an aliquot of the liquefied formulation is added to an open canister under conditions which are sufficiently cold to ensure the formulation does not vaporise, and then a metering valve crimped onto the canister.
  • each filled canister is check- weighed, coded with a batch number and packed into a tray for storage before release testing.
  • Suspensions and solutions comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof may also be administered to a patient via a nebulizer.
  • the solvent or suspension agent utilized for nebulization may be any pharmaceutically-acceptable liquid such as water, aqueous saline, alcohols or glycols, e.g., ethanol, isopropylalcohol, glycerol, propylene glycol, polyethylene glycol, etc. or mixtures thereof.
  • Saline solutions utilize salts which display little or no pharmacological activity after administration.
  • organic salts such as alkali metal or ammonium halogen salts, e.g., sodium chloride, potassium chloride or organic salts, such as potassium, sodium and ammonium salts or organic acids, e.g., ascorbic acid, citric acid, acetic acid, tartaric acid, etc. may be used for this purpose.
  • alkali metal or ammonium halogen salts e.g., sodium chloride, potassium chloride or organic salts, such as potassium, sodium and ammonium salts or organic acids, e.g., ascorbic acid, citric acid, acetic acid, tartaric acid, etc.
  • organic acids e.g., ascorbic acid, citric acid, acetic acid, tartaric acid, etc.
  • the compound of formula (I) or pharmaceutically acceptable salt thereof may be stabilized by the addition of an inorganic acid, e.g., hydrochloric acid, nitric acid, sulphuric acid and/or phosphoric acid; an organic acid, e.g., ascorbic acid, citric acid, acetic acid, and tartaric acid, etc., a complexing agent such as EDTA or citric acid and salts thereof; or an antioxidant such as antioxidant such as vitamin E or ascorbic acid. These may be used alone or together to stabilize the compound of formula (I) or pharmaceutically acceptable salt thereof.
  • an inorganic acid e.g., hydrochloric acid, nitric acid, sulphuric acid and/or phosphoric acid
  • an organic acid e.g., ascorbic acid, citric acid, acetic acid, and tartaric acid, etc.
  • a complexing agent such as EDTA or citric acid and salts thereof
  • an antioxidant such as antioxidant such as vitamin E or as
  • Preservatives may be added such as benzalkonium chloride or benzoic acid and salts thereof.
  • Surfactant may be added particularly to improve the physical stability of suspensions. These include lecithin, disodium dioctylsulphosuccinate, oleic acid and sorbitan esters.
  • the invention is directed to a dosage form adapted for intranasal administration.
  • Formulations for administration to the nose may include pressurised aerosol formulations and aqueous formulations administered to the nose by pressurised pump. Formulations which are non-pressurised and adapted to be administered topically to the nasal cavity are of particular interest. Suitable formulations contain water as the diluent or carrier for this purpose. Aqueous formulations for administration to the lung or nose may be provided with conventional excipients such as buffering agents, tonicity modifying agents and the like. Aqueous formulations may also be administered to the nose by nebulisation.
  • the compounds of formula (I) or pharmaceuticaily acceptable salts thereof may be formulated as a fluid formulation for delivery from a fluid dispenser, for example a fluid dispenser having a dispensing nozzie or dispensing orifice through which a metered dose of the fluid formulation is dispensed upon the application of a user-applied force to a pump mechanism of the fluid dispenser.
  • a fluid dispenser for example a fluid dispenser having a dispensing nozzie or dispensing orifice through which a metered dose of the fluid formulation is dispensed upon the application of a user-applied force to a pump mechanism of the fluid dispenser.
  • Such fluid dispensers are generally provided with a reservoir of multiple metered doses of the fluid formulation, the doses being dispensable upon sequential pump actuations.
  • the dispensing nozzle or orifice may be configured for insertion into the nostrils of the user for spray dispensing of the fluid formulation into the nasal cavity.
  • a fluid dispenser of the aforementioned type is described and illustrated in WO05/044354, the entire content of which is hereby incorporated herein by reference.
  • the dispenser has a housing which houses a fluid discharge device having a compression pump mounted on a container for containing a fluid formulation.
  • the housing has at least one finger-operable side lever which is movable inwardly with respect to the housing to cam the container upwardly in the housing to cause the pump to compress and pump a metered dose of the formulation out of a pump stem through a nasal nozzle of the housing.
  • the fluid dispenser is of the general type illustrated in Figures 30-40 of WO05/044354.
  • compositions adapted for intranasal administration wherein the carrier is a solid include a coarse powder having a particle size for example in the range 20 to 500 microns which is administered by rapid inhalation through the nasai passage from a container of the powder held close up to the nose.
  • suitable compositions wherein the carrier is a liquid, for administration as a nasal spray or as nasal drops include aqueous or oil solutions of the compound of formula (I) or a pharmaceutically acceptable salt thereof.
  • compositions adapted for transdermal administration may be presented as discrete patches intended to remain in intimate contact with the epidermis of the patient for a prolonged period of time.
  • the active ingredient may be delivered from the patch by iontophoresis as generally described in Pharmaceutical Research, 3(6), 318 (1986).
  • compositions adapted for topical administration may be formulated as ointments, creams, suspensions, lotions, powders, solutions, pastes, gels, sprays, aerosols or oils.
  • Ointments, creams and gels may, for example, be formulated with an aqueous or oily base with the addition of suitable thickening and/or gelling agent and/or solvents.
  • bases may thus, for example, include water and/or an oil such as liquid paraffin or a vegetable oil such as arachis oil or castor oil, or a solvent such as polyethylene glycol.
  • Thickening agents and gelling agents which may be used according to the nature of the base include soft paraffin, aluminium stearate, cetosteary!
  • Lotions may be formulated with an aqueous or oily base and will in general also contain one or more emulsifying agents, stabilising agents, dispersing agents, suspending agents or thickening agents.
  • Powders for externa! application may be formed with the aid of any suitable powder base, for example, talc, lactose or starch. Drops may be formulated with an aqueous or nonaqueous base also comprising one or more dispersing agents, solubilising agents, suspending agents or preservatives.
  • Topical preparations may be administered by one or more applications per day to the affected area; over skin areas occlusive dressings may advantageously be used. Continuous or prolonged delivery may be achieved by an adhesive reservoir system.
  • compositions may be applied as a topical ointment or cream.
  • the compound of formula (I) or a pharmaceutically acceptable salt thereof may be employed with either a paraffinic or a water-miscible ointment base.
  • the compound of formula (i) or pharmaceutically acceptable salt thereof may be formulated in a cream with an oil-in-water cream base or a water-in-oil base.
  • compositions adapted for parenteral administration include aqueous and non-aqueous sterile injection solutions which may contain anti-oxidants, buffers, bacteriostats and solutes which render the formulation isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions which may include suspending agents and thickening agents.
  • the compositions may be presented in unit- dose or multi-dose containers, for example sealed ampoules and vials, and may be stored in a freeze-dried (lyophilized) condition requiring only the addition of the sterile liquid carrier, for example water for injections, immediately prior to use.
  • Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules and tablets.
  • the compound and pharmaceutical formulations according to the invention may be used in combination with or include one or more other therapeutic agents, for example selected from anti-inflammatory agents, anticholinergic agents ⁇ particularly an bA M 2 /Mi receptor antagonist), 3 2 -adrenoreceptor agonists, antiinfective agents, such as antibiotics or antivirals, or antihistamines.
  • other therapeutic agents for example selected from anti-inflammatory agents, anticholinergic agents ⁇ particularly an bA M 2 /Mi receptor antagonist), 3 2 -adrenoreceptor agonists, antiinfective agents, such as antibiotics or antivirals, or antihistamines.
  • the invention thus provides, in a further aspect, a combination comprising a compound of formula (! or a pharmaceutically acceptable salt thereof together with one or more other therapeutically active agents, for example selected from an anti-inflammatory agent, such as a corticosteroid or an NSAID, an anticholinergic agent, a 2 -adrenoreceptor agonist, an antiinfective agent, such as an antibiotic or an antiviral, or an antihistamine.
  • an anti-inflammatory agent such as a corticosteroid or an NSAID
  • an anticholinergic agent such as a corticosteroid or an NSAID
  • an anticholinergic agent such as a corticosteroid or an NSAID
  • an anticholinergic agent such as a corticosteroid or an NSAID
  • an anticholinergic agent such as a corticosteroid or an NSAID
  • an anticholinergic agent such as a corticosteroid or an NSAID
  • One embodiment of the invention encompasses combinations comprising one or two other therapeutic agents.
  • the other therapeutic ingredient(s) may be used in the form of salts, for example as alkali metal or amine salts or as acid addition salts, or prodrugs, or as esters, for example lower alky! esters, or as solvates, for example hydrates to optimise the activity and/or stability and/or physical characteristics, such as solubility, of the therapeutic ingredient. It will be clear also that, where appropriate, the therapeutic ingredients may be used in optically pure form.
  • the invention encompasses a combination comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof together with a p 2 -adrenoreceptor agonist.
  • 2 -adrenoreceptor agonists examples include salmeterol (which may be a racemate or a single enantiomer such as the R-enantiomer), salbutamol (which may be a racemate or a single enantiomer such as the R-enantiomer), formoterol (which may be a racemate or a single duastereomer such as the R.
  • R-diastereomer salmefamol, fenoterol carmotero!, etanterol, naminterol, clenbuterol, pirbuterol, flerbuterol, reproteroi, bambuterol, indacaterol, terbutaline and salts thereof, for example the xinafoate (1 -hydroxy-2- naphthalenecarboxy!ate) salt of salmeterol, the sulfate salt or free base of salbutamol or the fumarate salt of formoterol.
  • long-acting p 2 -adrenoreceptor agonists for example, compounds which provide effective bronchodilation for about 12 hrs or longer, are preferred.
  • P 2 -adrenoreceptor agonists include those described in WO 02/066422, WO
  • Examples of ⁇ 2 ⁇ re no receptor agonists include:
  • the 3 2 -adrenoreceptor agonist may be in the form of a salt formed with a pharmaceutically acceptable acid selected from sulphuric, hydrochloric, fumaric, hydroxynaphthoic (for example 1- or 3-hydroxy-2-naphthoic), cinnamic, substituted cinnamic, triphenylacetic, sulphamic, sulphanilic, naphthaleneacrylic, benzoic, 4-methoxybenzoic, 2- or 4-hydroxybenzoic, 4-chlorobenzoic and 4-phenylbenzo ' ic acid.
  • a pharmaceutically acceptable acid selected from sulphuric, hydrochloric, fumaric, hydroxynaphthoic (for example 1- or 3-hydroxy-2-naphthoic), cinnamic, substituted cinnamic, triphenylacetic, sulphamic, sulphanilic, naphthaleneacrylic, benzoic
  • Suitable anti-inflammatory agents include corticosteroids.
  • Suitable corticosteroids which may be used in combination with the compounds of formula (I) or pharmaceutically acceptable salts thereof are those oral and inhaled corticosteroids and their pro-drugs which have anti-inflammatory activity. Examples include methyl prednisolone, prednisolone, dexamethasone, fluticasone propionate, 6a,9co-difluoro-11 p-hydroxy-16a- methyl-17a-[(4-methyl-1 ,3-thiazole-5-carbonyl)oxy]-3-oxo-androsta-1 ,4-diene-17 ⁇ - carbothioic acid S-fluoromethy!
  • ester (fluticasone furoate), 6a,9a-difluoro-1 1 -hydroxy-16a-methyl-3-oxo-17a-propionyloxy- androsta-1 ,4-diene-17p-carbothioic acid S-(2-oxo-tetrahydro-furan-3S-yl) ester, 6 ⁇ ,9 ⁇ - difluoro-11 -hydroxy-16a-methyl-3-oxo-17a-(2,2,3,3- tetramethycyciopropylcarbonyl)oxy- androsta-1 ,4-diene-17p-carbothioic acid S-cyanomethyl ester and 6a,9a-difluoro-1 i p- hydroxy-16a-methyl-17a-(1-methycyc!opropylcarbonyl)oxy-3-oxo-androsta-1 ,4-diene-17p- carbothioic acid S-fiuoromethyl ester, beclo
  • Preferred corticosteroids include fluticasone propionate, 6a,9a-difluoro-1 I p-hydroxy-iea-methyl- ⁇ a-t -methyl-l ⁇ -thiazoie-S-carbony oxyj-S-oxo-andro ⁇
  • the corticosteroid is 6 ⁇ ,9 ⁇ - difluoro-17cc-[(2-furanylcarbonyi)oxy]-1 1 p-hydroxy-16 -methyl-3-oxo-androsta-1 ,4-diene- 17p-carbothioic acid S-fiuoromethyl ester.
  • corticosteroids may include those described in WO2002/088167, WO2002/100879, WO2002/12265, WO2002/12266, WO2005/005451 , WO2005/005452, WO2006/072599 and WO2006/072600.
  • Non-steroidal compounds having glucocorticoid agonism that may possess selectivity for transrepression over transactivation and that may be useful in combination therapy include those covered in the following patents: WO03/082827, W098/54159, WO04/005229, WO04/009017, WO04/018429, WO03/104195, WO03/082787, WO03/082280, WO03/059899, WO03/101932, WO02/02565, WO01/16128, WO00/66590, WO03/086294, WO04/026248, WO03/061651 and WO03/08277. Further non-steroidal compounds are covered in: WO2006/000401 , WO2006/000398 and WO2006/015870.
  • anti-inf!ammatory agents examples include non-steroidal anti-inflammatory drugs (NSAID's).
  • NSAID's non-steroidal anti-inflammatory drugs
  • NSAID's examples include sodium cromoglycate, nedocromil sodium, phosphodiesterase (PDE) inhibitors (for example, theophylline, PDE4 inhibitors or mixed PDE3/PDE4 inhibitors), leukotriene antagonists, inhibitors of !eukotriene synthesis (for example montelukast), iNOS inhibitors, tryptase and elastase inhibitors, beta-2 integrin antagonists and adenosine receptor agonists or antagonists (e.g.
  • adenosine 2a agonists adenosine 2a agonists
  • cytokine antagonists for example chemokine antagonists, such as a CCR3 antagonist
  • inhibitors of cytokine synthesis or 5-lipoxygenase inhibitors.
  • An iNOS (inducible nitric oxide synthase inhibitor) is preferably for oral administration.
  • iNOS inhibitors include those disclosed in WO93/ 3055, WO98/30537, WO02/50021 , W095/34534 and W099/62875.
  • CCR3 inhibitors include those disclosed in WO02/26722.
  • the invention provides the use of the compounds of formula (I) in combination with a phosphodiesterase 4 (PDE4) inhibitor, especially in the case of a formulation adapted for inhalation.
  • PDE4-specific inhibitor useful in this aspect of the invention may be any compound that is known to inhibit the PDE4 enzyme or which is discovered to act as a PDE4 inhibitor, and which are only PDE4 inhibitors, not compounds which inhibit other members of the PDE family, such as PDE3 and PDE5, as well as PDE4.
  • Compounds include c/s-4-cyano-4-(3-cyclopentyloxy-4-methoxyphenyl)cyclohexan-1 - carboxylic acid, 2-carbomethoxy-4-cyano-4-(3-cyclopropylmethoxy-4- difluoromethoxyphenyl)cyclohexan-1-one and c/ ' s-[4-cyano-4-(3-cyclopropylmethoxy-4- difluoromethoxyphenyl)cyclohexan-1 -of].
  • anticholinergic agents are those compounds that act as antagonists at the muscarinic receptors, in particular those compounds which are antagonists of the or M 3 receptors, dual antagonists of the ⁇ ,/ ⁇ or M 2 /M 3 , receptors or pan-antagonists of the i/M 2 / 3 receptors.
  • exemplary compounds for administration via inhalation include ipratropium (for example, as the bromide, CAS 22254-24-6, sold under the name Atrovent), oxitropium (for example, as the bromide, CAS 30286-75-0) and tiotropium (for example, as the bromide, CAS 136310-93-5, sold under the name Spiriva).
  • revatropate for example, as the hydrobromide, CAS 262586-79-8) and LAS- 34273 which is disclosed in WO01/041 18.
  • Exemplary compounds for oral administration include pirenzepine (CAS 28797-61-7), darifenacin (CAS 133099-04-4, or CAS 133099- 07-7 for the hydrobromide sold under the name Enablex), oxybutynin (CAS 5633-20-5, sold under the name Ditropan), terodiline (CAS 15793-40-5), tolterodine (CAS 124937-51- 5, or CAS 124937-52-6 for the tartrate, sold under the name Detrol), otiionium (for example, as the bromide, CAS 26095-59-0, sold under the name Spasmomen), trospium chioride (CAS 10405-02-4) and solifenacin (CAS 242478-37-1 , or CAS 242478-38-2 for the succinate
  • anticholinergic agents include compounds which are disclosed in US patent application 60/511009 including, for example:
  • the invention provides a combination comprising a compound of formula (i) or a pharmaceutically acceptable salt thereof together with an H1 antagonist
  • H1 antagonists include, without limitation, amelexanox, astemizole, azatadine, azelastine, acrivastine, brompheniramine, cetirizine, levocetirizine, efietirizine, chlorpheniramine, clemastine, cyc!izine, carebastine, cyproheptadine, carbinoxamine, descarboethoxyloratadine, doxylamine, dimethindene, ebastine, epinastine, efietirizine,
  • the invention provides a combination comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof together with an H3 antagonist (and/or inverse agonist).
  • H3 antagonists include, for example, those compounds disclosed in WO2004/035556 and in WO2006/045416.
  • Other histamine receptor antagonists which may be used in combination with the compounds of the present invention include antagonists (and/or inverse agonists) of the H4 receptor, for example, the compounds disclosed in Jablonowski et a/., J. Med. Chem. 46:3957-3960 (2003).
  • the invention thus provides, in a further aspect, a combination comprising a compound of formula (!) or a pharmaceutically acceptable salt thereof together with a PDE4 inhibitor.
  • the invention thus provides, in a further aspect, a combination comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof together with a 2 -adrenoreceptor agonist.
  • the invention thus provides, in a further aspect, a combination comprising a compound of formula (!) or a pharmaceutically acceptable salt thereof together with a corticosteroid.
  • the invention thus provides, in a further aspect, a combination comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof together with a non-steroidal GR agonist.
  • the invention thus provides, in a further aspect, a combination comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof together with an anticholinergic.
  • the invention thus provides, in a further aspect, a combination comprising a compound of formula (!) or a pharmaceutically acceptable salt thereof together with an antihistamine.
  • the invention thus provides, in a further aspect, a combination comprising a compound of formula (I) or a pharmaceutically acceptable sa!t thereof together with a PDE4 inhibitor and a agonist.
  • the invention thus provides, in a further aspect, a combination comprising a compound of formula (! or a pharmaceutically acceptable salt thereof together with an anticholinergic and a PDE-4 inhibitor.
  • the combinations referred to above may conveniently be presented for use in the form of a pharmaceutical composition and thus pharmaceutical compositions comprising a combination as defined above together with a pharmaceutically acceptable diluent or carrier represent a further aspect of the invention.
  • the individual compounds of such combinations may be administered either sequentially or simultaneously in separate or combined pharmaceutical formulations.
  • the individual compounds will be administered simultaneously in a combined pharmaceutical formulation.
  • Appropriate doses of known therapeutic agents will readily be appreciated by those skilled in the art.
  • the invention thus provides, in a further aspect, a pharmaceutical composition
  • a pharmaceutical composition comprising a combination of a compound of formula (I) or a pharmaceutically acceptable salt thereof together with another therapeutically active agent.
  • the invention thus provides, in a further aspect, a pharmaceutical composition
  • a pharmaceutical composition comprising a combination of a compound of formula (I) or a pharmaceutically acceptable salt thereof together with a PDE4 inhibitor.
  • the invention thus provides, in a further aspect, a pharmaceutical composition comprising a combination of a compound of formula (I) or a pharmaceutically acceptable salt thereof together with a p 2 -adrenoreceptor agonist.
  • a pharmaceutical composition comprising a combination of a compound of formula (I) or a pharmaceutically acceptable salt thereof together with a corticosteroid.
  • the invention thus provides, in a further aspect, a pharmaceutical composition
  • a pharmaceutical composition comprising a combination of a compound of formula (I) or a pharmaceutically acceptable salt thereof together with a non-steroidal GR agonist.
  • the invention thus provides, in a further aspect, a pharmaceutical composition comprising a combination of a compound of formula (I) or a pharmaceutically acceptable salt thereof together with an anticholinergic.
  • a pharmaceutical composition comprising a combination of a compound of formula (I) or a pharmaceuticaily acceptable salt thereof together with an antihistamine.
  • a pharmaceutical composition comprising a combination of a compound of formula (i) or a pharmaceutically acceptable salt thereof together with a PDE4 inhibitor and a p 2 -adrenoreceptor agonist.
  • the invention thus provides, in a further aspect, a pharmaceutical composition
  • a pharmaceutical composition comprising a combination of a compound of formula (f) or a pharmaceuticaily acceptable salt thereof together with an anticholinergic and a PDE4 inhibitor.
  • A 0.1 % v/v solution of formic acid in water.
  • the UV detection was an averaged signal from wavelength of 210nm to 350nm and mass spectra were recorded on a mass spectrometer using alternate-scan positive and negative mode electrospray ionization.
  • A 10 mM ammonium bicarbonate in water adjusted to pH 10 with ammonia solution.
  • B acetonitrile.
  • the UV detection was an averaged signal from wavelength of 210nm to 350nm and mass spectra were recorded on a mass spectrometer using alternate-scan positive and negative mode electrospray ionization.
  • the UV detection was an averaged signal from wavelength of 210nm to 350nm and mass spectra were recorded on a mass spectrometer using alternate-scan positive and negative mode electrospray ionization.
  • A 10 mM ammonium bicarbonate in water adjusted to pH 10 with ammonia solution.
  • B acetonitrile.
  • the UV detection was an averaged signal from wavelength of 210nm to 350nm and mass spectra were recorded on a mass spectrometer using alternate-scan positive and negative mode electrospray ionization.
  • the UV detection was an averaged signal from waveiength of 210nm to 350nm and spectra were recorded on a mass spectrometer using alternate-scan positive and negative mode electrospray ionization.
  • A 0.1 % v/v solution of formic acid in water.
  • HPLC analysis was conducted on a Sunfire C18 column (150mm x 30mm interna! diameter, 5pm packing diameter) at ambient temperature.
  • A 0.1 % v/v solution of trifluoroacetic acid in water.
  • HPLC analysis was conducted on an XBridge C18 column (150mm x 30mm internal diameter, 5pm packing diameter) at ambient temperature.
  • A 10 mM ammonium bicarbonate in water adjusted to pH 10 with ammonia solution.
  • B acetonitrile.
  • the gradient employed was dependent upon the retention time of the particular compound undergoing purification as recorded in the analytical LCMS, and was as follows: For compounds with an analytical LCMS retention time below 0.6 minutes (LCMS method A) or below 1.5 minutes (LCMS method B) the following gradient was used: Time Flow Rate
  • the UV detection was an averaged signa! from wavelength of 210nm to 350nm and mass spectra were recorded on a mass spectrometer using alternate-scan positive and negative mode electrospray ionization.
  • a microwave vial was charged with A/-[6-[(methyloxy)methyl]-2-(methylsulfonyl)-4- pyrimidinyl][1 ,3]thiazolo[5,4-0]pyridin-2-amine (450 mg, 1.281 mmol) and 1 ,1 - dimethylethyl (2-aminoethyl)carbamate (308 mg, 1.921 mmol) in isopropanol (10 mL). The via! was sealed and was heated in a Biotage Initiator microwave system at 150 °C for 30 minutes. The residue was then evaporated to dryness.
  • a microwave vial was charged with 6-bromo-A/-(2-chioro-4-pyrimidinyl)-1 ,3-benzothiazol- 2-amine (281 mg, 0.823 mmol), 1 ,2-ethanediamine (0.165 mL, 2.468 mmol) and isopropanol (4 mL).
  • the microwave vial was sealed and was heated in a Biotage Initiator microwave to 150 °C for 45 minutes. After cooling, the solid was filtered under reduced pressure to give a yellow solid. The solid was transferred into a microwave vial. 1 ,2- Ethanediamine (0.165 mL, 2.468 mmol) and isopropanol (4 mL) were added.
  • a microwave vial was charged with A/-[6-[(methyloxy)methyl]-2-(methylsulfonyl)-4- pyrimidinyl][1 ,3]thia2olo[5,4-0]pyridin-2-amine (500 mg, 1.423 mmol), 1 ,1-dimethylethyl 4- amtno-1-piperidinecarboxylate (427 mg, 2.134 mmol) and isopropanol (5 mL).
  • the vial was sealed and was heated in a Biotage Initiator microwave at 150 °C for 45 minutes. After cooling, the reaction was filtered under reduced pressure to give a yellow solid.
  • a microwave vial was charged with /V-[6-[(methyioxy)methyl]-2-(methylsulfonyl)-4- pyrimidinyi][1 ,3]thiazolo[5,4-b]pyridin-2-amine (250 mg, 0.71 1 mmol), trans-4- aminocyclohexanol (246 mg, 2.134 mmol) and isopropanol (5 mL).
  • the vial was sealed and was heated in a Biotage Initiator microwave to 150 °C for 45 minutes. After cooling, the reaction was filtered under reduced pressure and the solid was washed with isopropanol. The filtrate was evaporated to dryness.
  • a microwave via! was charged with W-[6-[(methyloxy)methyl]-2-(methyisulfonyl)-4- pyrimidinyl][1 ,3]thiazolo[5,4-j ]pyhdin-2-amine (150 mg, 0.427 mmol), 1 , 1 -dimethylethyl (4- aminobutyl)carbamate (0.245 mL, 1.281 mmol) and isopropanol (4 mL).
  • the vial was sealed and was heated in a Biotage Initiator microwave at 150 °C for 45 minutes. After cooling, the reaction mixture was filtered under reduced pressure and the solid was washed with isopropanol.
  • a microwave vial was charged with A/-[6-[(methyloxy)methyl3-2-(methylsulfonyl)-4- pyrimidinyl3[1 ,3]thiazolo[5,4-d]pyridin-2-amine (150 mg, 0.427 mmol), 1 , 1 -dimethylethyi (3- aminopropyi)carbamate (0,224 mL, 1.281 mmol) and isopropanol (4 mL).
  • the vial was sealed and was heated in a Biotage Initiator microwave to 150 °C for 45 minutes. After cooling, the reaction was filtered under vacuum and the solid was washed with isopropanol to give the title compound (86 mg).
  • a microwave via! was charged with /V-[6-[(methyloxy)methyl]-2-(methylsulfonyl)-4- pyrimidinyl3[1 ,3]thiazolo[5,4-0]pyridin-2-amine (200 mg, 0.569 mmol), rrans-1 ,2- cyclohexanediamine (97 mg, 0.854 mmol) and isopropanol (4 mL).
  • the via! was sealed and was heated in a Biotage Initiator microwave at 150 °C for 30 minutes. The mixture was evaporated to dryness to give a yeliow solid.
  • a microwave vial was charged with A/-[6-[(methyloxy)methyi]-2-(methylsulfony!)-4- pyrimidinyl][1 ,3]thiazolo[5,4-j ]pyridin-2-amine (200 mg, 0.569 mmol), 1 ,3- cyclohexanediamine (97 mg, 0.854 mmol) and isopropanol (4 mL).
  • the vial was sealed and was heated in a Biotage Initiator microwave at 150 °C for 30 minutes. After cooling the reaction was filtered under reduced pressure and the solid was washed with isopropanol. The filtrate was evaporated to dryness to give a yeliow solid.
  • a microwave vial was charged with A/-[6-[(methyloxy)methyl]-2-(methylsu!fonyl)-4- pyrimidinyl][1 ,3]thiazolo[5,4-6]pyridin-2-amine (200 mg, 0.569 mmol), 1 , 1 -dimethylethyl (3S)-3-amino-1-pyrrolidinecarboxylate (159 mg, 0.854 mmol) and isopropanol (4 mL). The vial was sealed and was heated in a Biotage Initiator microwave at 150 °C for 30 minutes.
  • a microwave vial was charged with A/-[6-[(methyloxy)methyl]-2-(methylsulfonyi)-4- pyrimidinyl][1 ,3]thiazolo[5,4-i)]pyridin-2-amine (200 mg, 0.569 mmo!), 1 ,1 -dimethylethyl 1 - piperazinecarboxy!ate (159 mg, 0.854 mmol) and isopropanoi (4 mL).
  • the via! was sealed and was heated in a Biotage Initiator microwave at 150 °C for 30 minutes.
  • a microwave vial was charged with A/-[6-[(methyloxy)methyl]-2-(methylsulfonyl)-4- pyrimtdinyljt Sjthiazo!oiS ⁇ - ⁇ pyridin ⁇ -amine (200 mg, 0.569 mmoi), c/s-1 ,2- cyclohexanediamine (97 mg, 0.854 mmol) and isopropano! (4 mL).
  • the vial was sealed and was heated in a Biotage Initiator microwave at 150 °C for 30 minutes. The mixture was evaporated to dryness to give a ye!low solid.
  • a microwave vial was charged with /V-[6-[(methyloxy)methyl]-2-(methy!sulfonyl)-4- pyrimidinyl][1 ,3]thiazolo[5,4-0]pyridin-2-amine (200 mg, 0.569 mmol), trans- " ,A- cyclohexanediamine (65 mg, 0.569 mmol) and isopropano! (4 mL).
  • the vial was sealed and was heated in a Biotage initiator microwave at 50 °C for 30 minutes. After cooling, the reaction was filtered under reduced pressure and the solid was washed with isopropanol. The filtrate was evaporated to dryness to give a yellow solid.
  • a microwave vial was charged with A/-[6-[(methyloxy)methy!]-2 ⁇ (methylsulfonyi)-4- pyrimidinyl][1 ,3]thiazolo[5,4-i>3pyridin-2-amine (200 mg, 0.569 mmol), 1 , 1-dimethylethyl (3R)-3-amino-1-pyrrolidinecarboxylate (159 mg, 0.854 mmol) and isopropanol (4 mL). The vial was sealed and was heated in a Biotage Initiator microwave at 150 °C for 30 minutes. A further portion of 1 ,1-dimethylethy!
  • a microwave vial was charged with A/-[6-[(methyioxy)methyl]-2-(methylsulfony!)-4- pyrimidinyl][1 ,3]thiazolo[5,4-t>]pyridin-2-amine (200 mg, 0.569 mmol), 1 ,1 -dimethylethyl 4- hydroxy-1-piperidinecarboxylate (229 mg, 1.138 mmol) and sodium hydride (60 % w/w in mineral oil) (68.3 mg, 1.707 mmol) in tetrahydrofuran (3 ml). The vial was sealed and was heated in a Biotage Initiator microwave at 130 °C for 30 minutes.

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Abstract

The invention is directed to certain novel compounds. Specifically, the invention directed to compounds of formula (I) and salts thereof. The compounds of the invention are inhibitors of kinase activity, in particular Itk activity.

Description

NOVEL COMPOUNDS
FIELD OF THE INVENTION
The present invention is directed to certain novel compounds which are inhibitors of kinase activity, processes for their preparation, pharmaceutical compositions comprising the compounds, and the use of the compounds or the compositions in the treatment of various disorders. More specifically, the compounds of the invention are inhibitors of the activity or function of Itk (interleukin-2 inducible tyrosine kinase). Compounds which are inhibitors of the activity or function of Itk may be useful in the treatment of disorders such as respiratory diseases including asthma, chronic obstructive pulmonary disease (COPD) and bronchitis; allergic diseases including allergic rhinitis and atopic dermatitis; autoimmune diseases including rheumatoid arthritis, multiple sclerosis, psoriasis, type I diabetes, T cell mediated hypersensitivities, Guiliain-Barre Syndrome and Hashimoto's thyroiditis; transplant rejection; graft versus host disease; inflammatory disorders including conjunctivitis, contact dermatitis, inflammatory bowel disease and chronic inflammation; HIV; aplastic anemia; and pain including inflammatory pain.
BACKGROUND OF THE INVENTION
lnterleukin-2 inducible tyrosine kinase (Itk) is a non-receptor tyrosine kinase of the Tec family, which is also known as Tsk or Emt. Other members of the Tec fami!iy include: Tec, Btk, Txk and Bmx. The Tec family kinases are predominantly expressed in haematopoietic cells, however Bmx and Tec have a wider expression profile. The Tec famiiy kinases share a common domain structure: an amino-terminal pleckstrin homology (PH) domain (absent in Txk), a tec homology domain (containing one or two proline rich regions), followed by Src homology SH3 and SH2 domains, and a carboxy-terminal kinase domain. The PH domain binds to Ptdln(3,4,5)P3, and is responsible for locating the Tec kinase to the plasma membrane, whilst the PRR, SH3 and SH2 domains are involved in protein-interactions important in formation of the signalling complex.
Itk expression is restricted to T cells, NK and mast cells. Itk is the predominant Tec family kinase in naive T cells, which also express Txk and Tec. Upon activation via the T cell receptor or interleukin-2 (IL-2), the expression of Itk increases. There is some evidence that Itk is preferentially expressed in Th2 over Th1 ceils, in contrast to Txk which is present at higher levels in Th1 cells (1 ). Itk plays a key role in T cell receptor signalling. Itk is recruited to the plasma membrane through interaction with Ptdlns(3,4,5)P3, which is generated by P!3kinase. Itk forms a complex with several signalling and scaffold proteins including SLP76 and LAT. Itk is transphorphorylated by Lck. Activated Itk phosphoryiates PLCy, leading to the generation of lns(1 ,4,5)P3 (required for calcium flux within the cells) and diacylglycerol (activates members of the protein kinase C family and RAS guanyl-releasing protein. This results in the activation of mitogen-activated protein kinases (including JNK and ERK) and other effectors that regulate gene transcription, leading to the secretion of cytokines (reviewed in ref 2). In addition to the role of itk in PLCy activation and Ca2+ mobilisation, Itk may also contribute to TCR-induced actin reorganisation, and formation of the immune synapse. However, regulation of the actin cytoskeleton may not require kinase activity (3), suggesting the importance of Itk as a scaffold protein. In addition to the T cell receptor, itk may also be activated via the chemokine receptor CXCR4 (4) in T cells, and via the FceRI in mast cells (5).
There is considerable evidence suggesting that T cells play a key role in the pathogenesis of asthma. The inhibition of T cell cytokines will dampen down the inflammatory cascade involved in the asthmatic response. Cyclosporin A (CsA), which is thought to exert its major effect via inhibition of T cell cytokine release, has shown significant improvement in lung function in two trials with severe asthmatics (6,7). There is also evidence that CsA is steroid sparing and may lead to fewer exacerbations (7). A further trial reported some benefit of CsA but was non-significant (8). CsA does have actions on other cell types (e.g. mast cells) in addition to T cells. However, following allergen challenge in allergic asthmatics, CsA inhibited the late phase but not the early phase response (9), suggesting that effects on mast cells are unlikely to play a key role in the beneficial effect seen of CsA. Furthermore, daclizumab, an antibody against the anti-IL-2Ra chain (CD25) of activated lymphocytes improved pulmonary function and asthma control in patients with moderate to severe chronic asthma (10), supporting anti-T cell therapy for asthma.
Inhibition of Itk represents a potential novel therapy for asthma, by inhibiting T cell cytokine release. The key role for Itk in T cell receptor signalling has been demonstrated using Itk-/- mice and siRNA. In vitro activation of CD4+ cells from Itk knockout mice show reduced levels of Th2 (11) or both Th1 and Th2 (12) cytokines compared to wild type. Naive T cells from Itk knockout mice can differentiate normally into either Th1 or Th2 cells if cultured in vitro under appropriate cytokine conditions, suggesting that Itk is not required for Th2 cell differentiation (12). Studies differ in the reported effect of Itk knockout on cytokine release upon re-stimulation, showing either a selective reduction in Th2 or reduction in both Th1 and Th2 cytokines (12, 13). Itk siRNA inhibits cytokine release (Th1 and Th2) from human peripheral blood T cells following activation either with anti- CD3/CD28 or in response to recall antigen in vitro.
Itk-/- mice show reduced lung Th2 cytokine production, cell influx, mast ceil degranulation and airway hyperreactivity to methacholine in murine Ova challenge models (14, 15,16). In addition to these knockout studies there is also evidence that an itk inhibitor is effective at reducing cellular influx in an ova murine model of allergic asthma (17). These studies, together with the in vitro profile of Itk inhibitors in human T cells, suggests that Itk is a potential novel target for asthma therapy.
Inhibition of Itk may be beneficial in a variety of T-ce!l mediated diseases. In addition to asthma, Itk may play a role in other allergic diseases such as allergic rhinitis and atopic dermatitis. Single nucleotide polymorphisms in Itk have been associated with atopy (18) and seasonal ailergic rhinitis (19). itk mRNA levels in the peripheral blood T cells of atopic dermatitis patient is elevated in T cells from affected patients, compared to healthy controls (20).
References
1. Miller ef a/. Immunity (2004) 21 , 67-80
2. Schwartzberg et al. Nature Reviews Immunol (2005) 5, 284-295
3. Grasis et al. J Immunol (2003) 170, 3971-3976
4. Fischer et al. J Biol Chem (2004) 279, (28), 29816-29820
5. Kawakami et al. J Immunol (1995) 155, 3556-3562
6. Alexander ef al. The Lancet (1992) 339, 324-328
7. Lock et al. Am J Respir Crit Care Med (1996) 153, 509-514
8. Nizankowska et al. Eur Respir J (1995) 8, 1091 -1099
9. Sihra ef a/.Thorax (1997) 52, 447-452
10. Busse et al. Am J Respir Crit Care med (2008) 178, 1002-1008
11. Fowell et al. Immunity (1999) 11 , 399-409
12. Schaeffer ef al. Nature Immunol (2001 ) 2, (12) 1183-1 188
13. Au-Yeung et al. J Immunol (2006) 176, 3895-3899
14. Ferrara ef al. J Allergy Clin Immunol (2006) 117, 780-786
15. Forsseil ef al. Am J Respir Cell Mol Biol (2005) 32, 511 -520
16. Mueller and August J Immunol (2003) 170, 5056-5063
17. Lin et al. Biochemistry (2004) 43, (34) 1 1056-1 1062
18. Graves ef al. J Allergy Clin Immunol (2005) 116, 650-656 19. Benson ef al. Ailergy (2009) 64, 1286-1291 (DO! :10.1 1 1398- 9995.2009.01991.x)
20. Matsumoto ef al. Int Arch A!lergy Immunol (2002) 129, 327-340 Attempts have been made to prepare compounds which inhibit Itk activity and a number of such compounds have been disclosed in the art. However, in view of the number of pathological responses which are mediated by Itk, there remains a continuing need for inhibitors of Itk which can be used in the treatment of a variety of conditions. The present inventors have discovered novel compounds which are inhibitors of kinase activity, in particular Itk activity, and which may have the potential for irreversible inhibition. Compounds which are Itk inhibitors may be useful in the treatment of disorders associated with inappropriate kinase activity, in particular inappropriate Itk activity, for example in the treatment and prevention of disorders mediated by itk mechanisms. Such disorders include respiratory diseases including asthma, chronic obstructive pulmonary disease (COPD) and bronchitis; allergic diseases including allergic rhinitis and atopic dermatitis; autoimmune diseases including rheumatoid arthritis, multiple sclerosis, psoriasis, type I diabetes, T cell mediated hypersensitivities, Guil!ain-Barre Syndrome and Hashimoto's thyroiditis; transplant rejection; graft versus host disease; inflammatory disorders including conjunctivitis, contact dermatitis, inflammatory bowel disease and chronic inflammation; HIV; aplastic anemia; and pain including inflammatory pain.
In one embodiment, compounds of the invention may show selectivity for itk over other kinases.
In another embodiment, compounds of the invention may be potent inhibitors of Itk activity.
In another embodiment, compounds of the invention may be irreversible inhibitors of Itk activity.
In a further embodiment, compounds of the invention may show a fast rate of irreversible inhibition of Itk activity.
SUMMARY OF THE INVENTION
The invention is directed to certain novel compounds. Specifically, the invention is directed to compounds of formula (i):
Figure imgf000006_0001
(I)
wherein R1 to R3, X and Y are as defined below, and salts thereof.
The compounds are inhibitors of kinase activity, in particular Itk activity, and may have the potential for irreversible inhibition. Compounds which are Itk inhibitors may be useful in the treatment of disorders associated with inappropriate Itk activity, such as asthma. Accordingly, the invention is further directed to pharmaceutical compositions comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof. The invention is still further directed to methods of inhibiting Itk activity and treatment of disorders associated therewith using a compound of formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising a compound of formula (I) or a pharmaceuticaliy acceptable salt thereof. The invention is yet further directed towards processes for the preparation of the compounds of the invention.
BRIEF DESCRIPTION OF THE FIGURES
Figure 1 shows an X-ray powder diffraction (XRPD) pattern for A/-[(3S)-1 -acryloyl-3- pyrrolidinyl]-W-[1 ,3]thiazolo[5,4-b]pyridin-2-yl-4-({[(1 S)-1 ,2,2- trimethylpropyl]amino}methyi)-2,6-pyridinediamine difumarate monohydrate.
DETAILED DESCRIPTION OF THE INVENTION
in one embodiment, the invention is directed to compounds of formula (I)
Figure imgf000006_0002
(I)
wherein R1 is hydrogen, -CH2OR4, -CH2NR5R6, -CH2-pheny[ wherein ihe phenyl is optionally substituted by one or two substituents independently selected from Chalky!, C1-6alkoxy, nitrile and halo, -CH2-6-membered heterocyclyl wherein the 6-membered heterocyclyl optionally contains an oxygen atom and is optionally substituted by one or two substituents independently selected from Ci„6aSkyl and halo, or -CH2-5-membered heteroaryl wherein the 5-membered heteroaryl contains one or two nitrogen atoms and is optionally substituted by C1-6alkyl;
R2 is -(NH)2COCHCH2> -NR7R8, -OR9 or phenyl wherein the phenyl is optionally substituted by -NR10R1 1 ;
R3 is hydrogen, halo, C-|.6alkyl or C^alkoxy;
R6 and R6 are each independently hydrogen or C^alkyl, or
R5 and R6, together with the nitrogen atom to which they are attached, are linked to form a
5- or 6-membered heterocyclyl wherein the 6-membered heterocyclyl optionally contains an oxygen atom or a further nitrogen atom and the 5- or 6-membered heterocyclyl is optionally substituted by one or two substituents independently selected from C1-6alky! and halo;
R7 is hydrogen or methyl,
R8 is -COCHCH2l -{CH2)nNR12R13, C3.6cycloalkyl substituted by -OS02CH3 or -NR14R1S, or 4-, 5- or 6-membered heterocyclyl wherein the 4-, 5- or 6-membered heterocyclyl contains a nitrogen atom and is substituted by -R16, or
R7 and R8, together with the nitrogen atom to which they are attached, are linked to form a
6- membered heterocyclyl optionally containing a further nitrogen atom wherein the 6- membered heterocyclyl is substituted by -R17 or -NR18R19;
R9 is 5- or 6-membered heterocyclyl wherein the 5- or 6-membered heterocyclyl contains a nitrogen atom and is substituted by -R20;
R1 1 , R13, R15, R 6, R17, R19 and R20 are each independently -COCR25CR21 R26, -COCH2halo or -COCH2OR22; R2 is hydrogen, methyl, -CF3 or -CH2NR23R24;
R22 is phenyl optionally substituted by halo; R4, R10, R 2, R14, R18, R23 and R24 are each independently hydrogen or Chalky!; R25 is hydrogen, methyl or -CH2NR27R28;
R26 is hydrogen or methyl; and R , together with the nitrogen atom to which they are attached, are linked to form a 6-membered heterocyclyl optional!y containing an oxygen atom; X and Y are each independently -N- or -CH-; and n is 2, 3 or 4; and salts thereof (hereinafter "compounds of the invention").
As the skilled person will appreciate, groups such as R2 is -(NH)2COCHCH2, R8 is - COCHCH2l and R11, R13, R 5, R16, R 7, R19 and R20 are each independently - COCR25CR21R26, contain carbon-carbon double bonds and may be depicted as shown below wherein "-" denotes the point of attachment:
Figure imgf000008_0001
In one embodiment, R is hydrogen, ~CH2OR4, -CH2NR5RS, -CH2-phenyl wherein the phenyl is optionally substituted by one or two substituents independently selected from 6alkyl, C 6a!koxy, nitrilo and halo, or -CH2-5-membered heteroaryl wherein the 5- membered heteroaryl contains one or two nitrogen atoms and is optionally substituted by C1-6alkyl. In another embodiment, R1 is hydrogen, -CH2OR4, -CH2NR5R6 or -CH2-5- membered heteroaryl wherein the 5-membered heteroaryl contains one or two nitrogen atoms and is optionally substituted by Chalky!. In another embodiment, R1 is hydrogen, - CH2OR4, -CH2-6-membered heterocyclyl wherein the 6-membered heterocyclyl optionally contains an oxygen atom and is optionally substituted by one or two substituents independently selected from Chalky! and halo, -CH2-5-membered heteroaryi wherein the heteroaryi contains one or two nitrogen atoms and is optionally substituted by Chalky!. In another embodiment, R1 is -Ch^OR4, or -CH2-6-membered heterocyciyl wherein the 6- membered heterocyciyl optionally contains an oxygen atom and is optionally substituted by one or two substituents independently selected from C1-6alkyl and halo. In another embodiment, R1 is -CH2OR4, in particular wherein R4 is methyl, !n another embodiment, R1 is -CH2-6-membered heterocyciyl wherein the 6-membered heterocyciyl optionally contains an oxygen atom and is optionally substituted by one or two substituents independently selected from C1-6alky! and halo, in particular N-morpho!inyl. In a further embodiment, R1 is -CH2NR5R6.
In one embodiment, R2 is -NR7R8, -OR9 or phenyl wherein the phenyl is optionally substituted by -NR10R11.
In one embodiment, R2 is -NR7R8.
In one embodiment, R2 is -NR7R8 wherein R7 is hydrogen. In one embodiment, R2 is -NR7R8 wherein R8 is -(CH2)nNR12R13.
In one embodiment, R2 is -NR7R8 wherein R8 is C3.6cycloalkyl substituted by -NR14R15, in particular C6cycloalkyl substituted by -NR14R15. In one embodiment, R2 is -NR7R8 wherein R8 is 5- or 6-membered heterocyciyl wherein the 5- or 6-membered heterocyciyl contains a nitrogen atom and is substituted by -R16.
In one embodiment, R2 is -NR7R8 wherein R8 is 5-membered heterocyciyl wherein the 5- contains a nitrogen atom and is substituted by -R16.
In one embodiment, R12 or R14 are each independently hydrogen.
In one embodiment, R13, R15, or R16 are each independently -COCHCHR21 wherein R21 is hydrogen, -CF3 or -CH2N(CH3)2, in particular -COCHCH2 (R21 = hydrogen).
In another embodiment, R2 is -NR7R8 wherein R7 and R8, together with the nitrogen atom to which they are attached, are linked to form a 6-membered heterocyciyl optionally containing a further nitrogen atom wherein the 6-membered heterocyciyi is substituted by -R 7 or -NR18R19; and wherein R16 is hydrogen and R17 or R19 are each independently - COCHCHR21 wherein R21 is hydrogen, -CF3 or -CH2N(CH3)2, in particular -COCHCH2 (R21 = hydrogen).
In one embodiment, R3 is hydrogen or halo, for example bromo. In another embodiment, R3 is hydrogen. In a further embodiment, R3 is C1-6alkoxy, for example methoxy. in one embodiment, R4 is Chalky!, for example methyl.
In one embodiment, Rs and R6 are each independently hydrogen or Chalky!. In another embodiment, R5 is hydrogen and Re is C1-6alkyi, for example 3,3-dimethylbutan-2-yl- (1 ,2,2-trimethylpropyl-). In a further embodiment, R5 and R6 are each independently C-,. 6alkyl, for example methyl.
In one embodiment, R5 and R6, together with the nitrogen atom to which they are attached, are linked to form a 5- or 6-membered heterocyciyi wherein the 6-membered heterocyciyi optionally contains an oxygen atom or a further nitrogen atom and the 5- or 6- membered heterocyciyi is optionally substituted by one or two substituents independently selected from C -6alkyl and halo. In another embodiment, R5 and R6, together with the nitrogen atom to which they are attached, are linked to form a 5- or 6-membered heterocyciyi wherein the 6-membered heterocyciyi optionally contains an oxygen atom or a further nitrogen atom and the 5- or 6-membered heterocyciyi is optionally substituted by one or two substituents independently selected from d-ealkyl. in another embodiment, R5 and R6, together with the nitrogen atom to which they are attached, are linked to form a 6- membered heterocyciyi wherein the 6-membered heterocyciyi optionally contains an oxygen atom and is optionally substituted by one or two substituents independently selected from C^alkyl and halo. In a further embodiment, R5 and R6, together with the nitrogen atom to which they are attached, are linked to form a 6-membered heterocyciyi wherein the 6-membered heterocyciyi contains an oxygen atom and is substituted by two substituents independentiy selected from Chalky!, for example methyi.
In one embodiment, R7 is hydrogen or methyl,
R8 is -COCHCH2, -(CH2)nNR12R13, C3.6cycloalkyi substituted by -OS02CH3 or -NR1"R15, or 5- or 6-membered heterocyciyi wherein the 5- or 6-membered heterocyciyi contains a nitrogen atom and is substituted by -R16, or R7 and R8, together with the nitrogen atom to which they are attached, are linked to form a 6-membered heterocyclyl optionally containing a further nitrogen atom wherein the 6- membered heterocyclyl is substituted by -R 7 or -NR18R19. In one embodiment, R7 is hydrogen.
In one embodiment, R8 is 5- or 6-membered heterocyclyl wherein the 5- or 6-membered heterocyclyl contains a nitrogen atom and is substituted by -R16. In a further embodiment, R8 is 5-membered heterocyclyl wherein the 5-membered heterocyclyl contains a nitrogen atom and is substituted by -R16.
In one embodiment, R9 is 6-membered heterocyclyl wherein the 6-membered heterocyclyl contains a nitrogen atom and is substituted by -R20. In one embodiment, R10 is hydrogen.
In one embodiment, R1\ R13, R15, R16, R17, R19 and R20 are each independently - COCHCHR21, -COCH2halo or -COCH2OR22. In one embodiment, R11 -COCHCHR21.
In one embodiment, R12 is hydrogen. In a further embodiment, R12 is Ci.6alkyl, for example methyl. In one embodiment, R 3 is -COCHCHR21. in one embodiment, R 4 is hydrogen.
In one embodiment, R15 is -COCHCHR21.
In one embodiment, R16 is -COCHCHR21, -COCH2halo or -COCH2OR2Z. In a further embodiment, R16 is -COCHCHR21.
In one embodiment, R 7 is -COCHCHR1 in one embodiment, R1s is hydrogen. In one embodiment, R19 is -COCHCHR21.
In one embodiment, R20 is -COCHCHR21. In one embodiment, R21 is hydrogen, -CF3 or -CH2NR23R24. In a further embodiment, R21 is hydrogen. in one embodiment, R22 is phenyl optionally substituted by fluoro. in one embodiment, R23 is C1-6aikyl, for example methyl. In one embodiment, R24 is C^alkyl, for example methyl. In one embodiment, R25 is hydrogen.
In one embodiment, R26 is hydrogen.
In one embodiment, R27 and R28, together with the nitrogen atom to which they are attached, are linked to form morpho!inyl.
In one embodiment, X is -N-. In a further embodiment, X is -CH-.
In one embodiment, Y is -N-. In a further embodiment, Y is -CH-. In one embodiment, n is 2.
It is to be understood that the present invention covers all combinations of substituent groups described hereinabove. In another embodiment, the invention is directed to compounds of formula (IA)
Figure imgf000012_0001
(IA)
wherein
R1 is hydrogen, -CH2OR4, -CH2NR5R6, -CH2-phenyl wherein the phenyl is optionally substituted by one or two substituents independently selected from Chalky! , C^alkoxy, nitrilo and halo, -CHr6-membered heterocyciyi wherein the 6-membered heterocydyi optionally contains an oxygen atom and is optionally substituted by one or two substituents independently selected from C^alkyl and halo, or -CH2-5-membered heteroary! wherein the 5-membered heteroaryl contains one or two nitrogen atoms and is optionally substituted by Chalky!;
R2 is -(NH)2COCHCH2, -NR7R8, -OR9 or phenyl wherein the phenyl is optionally substituted by -NR10R11; R3 is hydrogen, halo, Chalky!, C -6alkoxy;
R5 and R6 are each independently hydrogen or C1-6aikyl, or
R5 and R6, together with the nitrogen atom to which they are attached, are linked to form a 5- or 6-membered heterocydyi wherein the 6-membered heterocyciyi optionally contains an oxygen atom or a further nitrogen atom and the 5- or 6-membered heterocydyi is optionally substituted by one or two substituents independently selected from Chalky! and halo;
R7 is hydrogen or methyl,
R8 is -COCHCH2] ~(CH2)nNR1 R13, C3-6cycloalkyi substituted by -OS02CH3 or -NR1 R 5, or
5- or 6-membered heterocydyi wherein the 5- or 6-membered heterocydyi contains a nitrogen atom and is substituted by -R16, or
R7 and R8, together with the nitrogen atom to which they are attached, are linked to form a
6- membered heterocydyi optionally containing a further nitrogen atom wherein the 6- membered heterocyciyi is substituted by -R17 or -NR18R19;
R9 is 5- or 6-membered heterocyciyi wherein the 5- or 6-membered heterocyciyi contains a nitrogen atom and is substituted by -R20; R11, R13, R 5, R16, R17, R19 and R20 are each independently -COCHCHR21, -COCH2haio or - COCH2OR22; R21 is hydrogen, -CF3 or -CH2NR23R24; R22 is phenyl optionally substituted by halo; R4, R10, R12, R14, R 8, R23 and R24 are each independently hydrogen or Chalky!; X and Y are each independently -N- or -CH-; and n is 2, 3 or 4; and salts thereof.
In another embodiment, the invention is directed to compounds of formula (IB)
Figure imgf000014_0001
(IB)
R1 is hydrogen, -CH2OR4, -CH2NR5R6, -CH2-phenyl wherein the phenyl is optionally substituted by one or two substituents independently selected from Ci-6alkyl, C1-6alkoxy, nstrilo and halo, -CH2-6-membered heterocyclyl wherein the 6-membered heterocyclyl optionally contains an oxygen atom and is optionally substituted by one or two substituents independently selected from C1-6alkyl and halo, or -CH2-5-membered heteroaryl wherein the 5-membered heteroaryl contains one or two nitrogen atoms and is optionally substituted by C1-6alkyl;
R2 is -NR7R8, -OR9 or phenyl wherein the phenyl is optionally substituted by -NR10R1 1 ; R3 is hydrogen, halo, C1-6alkyl, C^alkoxy;
R5 and R6, together with the nitrogen atom to which they are attached, are linked to form a 6-membered heterocycly! wherein the 6-membered heterocyclyl optionally contains an oxygen atom and is optionaliy substituted by one or two substituents independently selected from C^alkyl and halo;
R7 is hydrogen or methyl,
R8 is -(CH2)nNR12R13, C3.6cycloalkyl substituted by -OS02CH3 or -NR14R15, or 5- or 6- membered heterocyciyi wherein the 5- or 6-membered heterocyciyi contains a nitrogen atom and is substituted by -R16, or
R7 and R8, together with the nitrogen atom to which they are attached, are linked to form a 6-membered heterocyciyi optionally containing a further nitrogen atom wherein the 6- membered heterocyciyi is substituted by -R17 or -NR18R19;
R9 is 5- or 6-membered heterocycly! wherein the 5- or 6-membered heterocyciyi contains a nitrogen atom and is substituted by -R20;
R 1, R13, R15, R16, R17, R19 and R20 are each independently -COCHCHR21, -COCH2halo or - COCH2OR22;
R21 is hydrogen, -CF3 or -CH2NR23R24;
R22 is phenyl optionally substituted by halo;
R4, R1Q, R12, R14, R18, R23 and R24 are each independently hydrogen or C^alkyl; X and Y are each independently -N- or -CH-; and n is 2, 3 or 4; and salts thereof.
In another embodiment, the invention is directed to compounds of formula (IC)
Figure imgf000015_0001
(IC)
wherein R is -CH2NR5R6;
R2 is -NR7R8;
R3 is hydrogen;
R5 and R6 are each independently hydrogen or C1-6alkyl, or
R5 and R6, together with the nitrogen atom to which they are attached, are linked to form a 6-membered heterocyclyl wherein the 6-membered heterocyclyl contains an oxygen atom and is substituted by two substituents independently selected from C1-ealkyl;
R7 is hydrogen;
R8 is 5-membered heterocyclyl wherein the 5-membered heterocyclyl contains a nitrogen atom and is substituted by -R 6;
R16 is -COCHCHR21; R21 is hydrogen; X is -N-; and Y is -CH-; and salts thereof.
!n a further embodiment, the invention is directed to compounds of formula (ID)
Figure imgf000016_0001
(ID)
wherein R1 is -CH2NR5R6; R2 is -NR7Re; R3 is hydrogen;
R5 and R6 are each independently hydrogen or C1-6alkyl; R7 is hydrogen; R8 is 5-membered heterocyciyi wherein the 5-membered heterocyciyi contains a nitrogen atom and is substituted by -R16;
R 6 is -COCHCHR21; R21 is hydrogen;
X is -N-; and
Y is -CH-; and salts thereof.
Compounds of the invention include the compounds of Examples 1 to 102 and salts thereof.
In one embodiment, the compound of the invention is selected from the group consisting of:
W-(2-{[4-[(methyloxy)methyi]-6-([1 ,3]thiazolo[5i4-0]pyridin-2-ylamino)-2- pyrimidinyl]amino}ethyl)-2-propenamide;
A/-[2-{{4-[(6-bromo-1 ,3-benzothiazol-2-yl)amino]-2-pyrimidinyl}amino)ethyl]-2- propenamide; ira 7s-4-{[4-(4-morpholinylmethyl)-6-([1 ,3]thiazolo[5,4-63pyridin-2-ylamino)-2- pyrimidiny!]amino}cyc!ohexyl methanesulfonate;
W2-(1 -acryloyl-3-piperidinyl)-6-[(methyloxy^
pyrimidinediamine;
A/ -(1-acryloyl-4-piperidinyl)-6-[(methyloxy)^
pyrimidinediamine;
/rans^-i^-EimethyloxyJmethylj-e-ill .SlthiazoloES^-^p ridin^-ylamino)^- pyrimidinyi]amino}cyciohexyl methanesulfonate;
A/-(4-{[4-[(methyloxy)rnethyl]-6-([1 ,3]thiazo!o[5J4-03pyridin-2-ylamino)-2- pyrimidinyl3amino}butyi)-2-propenamide;
A/-(3-{[4-E(methyloxy)methyl)-6-([1 ,3]th!azolo[5,4-63pyridin--2-ylarnino)-2- pyrimidinyl]amino}propy!)-2-propenamide;
A -(irai7S-2-{[4-[(methyloxy)methyl3-6-([1 ,3]thiazolo[5,4-03pyridin-2-ylamino)-2- pyrimidinyl]amino}cyclohexyl)-2-propenamide;
A/-(c/s-3-{[4-[(methyioxy)methyl]-6-([1 ,3]thiazoio[5l4-/>]pyridin-2-ylamino)-2- pyrimidinyl]amino}cyclohexyl)-2-propenamide;
/V-(irai?s-3-{[4-[(methyloxy)methyl3-6-([1 ,33thiazolot5,4-i)3pyridin-2-ylamtno)-2- pyrimidinyl]amino}cyclohexyi)-2-propenamide;
N (3S)-1-acry[oyl-3-pyrroiidinyl3-6-[(methyloxy^
2,4-pyrimidinedsamine;
A/-{2-(4-acryloyl-1 -piperazinyl)-6-E(rnethyloxy)methy]]-4-pyrimidinyl}[1 ,3]thiazolo[5,4- i>]pyridin-2-amine;
A/.(c s-2-{[4-[(methyloxy)methyl]-6-([1 ,3]thiazoio[5,4-i)]pyridin-2-ylamino)-2- pyrimidinyi]amino}cyclohexyl)-2-propenamide;
A/-(4-{[4-[(methyioxy)methyl]-6-([1 ,3]thiazolo[5,4-/b3pyndin-2-ylamino)-2- pyrimidiny!]amino}cyclohexy!)-2-propenamide;
A -(/ra/7S-4-{[4-[(methyioxy)methyl]-6-([1 ,33thiazolo[5,4-b]pyridin-2-ylamino)-2- pyrimidiny!]amino}cyclohexyl)-2-propenamide;
W -[(3R)-1 -acryloyl-3-pyrrolidinyl]-6-[(met^
2,4-pyrimidinediamine;
W-{2-[(1-acryloyl-4-piperidinyi)oxy]-6-[(methyto^
£>]pyridin-2-amine;
(2E)-4-(dimethy!amino)-A/-(2-{[4-[(methyioxy)methyl]-6-([1 ,33thiazolo[5,4-i)]pyndin-2- ylamino)-2-pyrimidinyl3amino}eihy!)-2-butenamide;
A/^1-[(2-r)-4-(dimethylamino)-2-butenoyi]-4-piperidinyl}-6-E(methyloxy)methyl]-W4- [ 133thiazolo[5,4-6]pyridin-2-yi-2J4-pyrimidinediamSne; W2-{1-[{2£)-4-(dimeihy!amino)-2-butenoyi]-3-piperidiny!}-6-[(methyloxy)m [I .SjthiazolotS^-bjpyridin^-yl^^-pyrimidinediamine;
/V-(2-{f4-(4-morphoiinylmeihyi)-6-([ i3]thiazo!o[5,4-0]pyridin-2-ylarriino)-2- pyrimidinyl]amino}ethyl)-2-propenamide;
W-ifrans^-^^-morpholinyimeihylJ-e-^l ^lthiazoiotS^-^pyridin^-ylamino)^- pyritnidinyl]amino}cyclohexyi)-2-propenamide;
N2-(1-acryloyl-3-piperidinyl)-6-(4-morpho^
pyrimidinediamine;
/V-(ira 7s-4-{[4-(4-morpholiny!methyl)-6-([1 l33thiazoio[5,4-jbjpyridin-2-ylamino)-2- pyrimidinyl]amino}cyclohexyl)-2-propenamide;
A/2-[(3R)-1-acryloyl-3-pyrrolidiny!]-6-(4-^^
yl-2,4-pyrimidinediarriine;
W-(c/s-4-{[4-(4-morpholinylmeth^^
pyrimldiny!]amino}cyclohexyl)-2-propenamide;
/V2-(1-acryloyl-4-piperidinyl)-6-(4-morphoiin^^
pyrimidinediamine;
N2-[(3S)-1-acryloyl-3-pyrrolidiny!]-6-^^
yi-2,4-pyrimidinediamine;
A/-(c s-2-{[4-(4-morphoiinylmethyl)-6-([1 ,3]thiazolo[5l4-0]pyridin-2-yiamin
pyrimidiny[]amino}cyclohexyl)-2-propenamide;
A/-(1-{6-t(6-bromo-1 ,3-benzothiazol-2-yl)amino]-4-[(2-methyl-1 H-imidazol-1-yl)meihyl]-2- pyridinyl}-3-piperidiny!)-2-propenamide;
Ay-[2-(4-acryloyl-1-piperaziny!)-6-(4-morphol^
/b]pyridin-2-amine;
W-[2-[(1-acryloyl-4-piperidinyl)oxy]-6-(4-mo
6]pyridin-2-amine;
/V-(c/s-3-{[4-(4-morpholinylmeihyl)-6-([1 ,3]t iazolo[5l4-0]pyridin-2-ylamino)-2- pyrimidinyl]amino}cyclohexyl)-2-propenamide;
W-(ira ?s-3-{t4-(4-morpholinylmethyl)-6-([1 I3]thiazo!o[5^-^]pyridin-2-ylamino)-2- pyrimidinyl]amino}cyclohexyl)-2-propenamide;
A/2-[1 -{chloroacetyl)-3-piperidinyl]-6-(4-m
yl-2,4-pyrimidinediamine;
/V-[2-(2-acry(oyitetrahydro-1 (2H)-pyridazinyl)-6-(4-morpholinylmeihyl)-4- pyrimidinyi][1 ,3]thiazo!o[5,4-0]pyndin-2-amine;
A 2-(1 -{[(4-fiuorophenyl)oxy3acetyl}-3-piperidinyl)-6-(4-morpho!inylmethyi)-A/''- [1 ,3]thiazolo[5,4-0]pyridin-2-yi-2,4-pyrimidinediamine; 2-chloro-/V-(2-{[4-(4-morphoiiny[methyl)-6-([1 >33thiazolo[5,4-b]pyridin-2-ylamino)-2- pyrimidinyl]amino}ethyl)acetamide;
{2£)-4,4^-trifiuoro-/V 2-{[4-(4-morpholinylmeihy!)-6-([1 ,3]thiazo!o[5l4-b]pyridin-2- ylamino)-2-pyrimidinyl]amino}ethyi)-2-butenamide;
2-[(4-ffuorophenyi)oxy]-A/-(2-{[4-(4-morphol!ny!methyl)-6-([1 ,3]thiazoio[5,4-^3pyndin-2- ylatnino)-2-pyrimidinyl]amino}ethyl)acetamide;
W-{1-f4-(4-morphoiinylmethyl)-6-([1 >3]thiazoio[5I4-0]pyridin-2-ylamino)-2-pyridi^ piperidinyi}-2-propenamide;
/V-(c/s-3-{[4-(4-morpholinyimethyl)-6^
pyridinyl]amino}cyc!ohexy!)-2-propenamide;
W-(ira/7S-3-{[4-{4-morpholinylmethyl)-6-([1 ,3]thiazo[o[5^-^]pyridin-2-yiamino)-2- pyridinyl]amino}cyc!ohexy!)-2-propenamide;
A/-(2-{[4-(4-morpholinylmethyl)-6-(E1 ,3]thiazoio[514-a]pyridin-2-ylamino)-2- pyridinyl]amino}ethyi)-2-propenamide;
W-c/s-2-{[4-(4-morpholiny[methyl)-6-([1 ,3]thiazolo[5,4-i)]pyridin-2-y!amino)-2- pyridinyl]amino}cyciohexyl)-2-propenamide;
A/-(ira/?s-2-{[4-(4-morpholinyimethyl)-6-([1 l33thiazo[o[5,4-jb]pyndin-2-ylamino)-2- pyridinyi]amino}cyclohexyi)-2-propenamide;
6-(4-morpho!inylmethyl)-W4-[113]ihtazolo[5,4-6]pyridin-2-yS-W2-{1 -[(2£)-4,4,^ butenoyi]-3-piperidinyl}-2,4-pyrimidinediamine;
W-me†hy^-(2-{[4-(4-morpholinylmethyl^
pyrimidinyl]amino}ethyi)-2-propenamide;
A/-{3-[4-{4-morphoiinylmethyl)-6-(t1 ,3]thiazo!o[5,4-J ]pyridin-2-ylamino)-2- pyrimidinyl]phenyl}-2-propenamide;
W-[4-(4-morpholinyimethyl)-6-([1 ,3]thiaz
propenamide;
W 4-(4-morphoiinylmethyt)-6-([1 ,33thiazoio[5,4-/b3pyridin-2-ylamino)-2(1 H)- pyrimidinylidene]-2-propenohydrazide;
/V2-[(3S)-1 -acryloyi-3-pyrrolidiny!]-A/"-(6-bromo-1 ,3-benzoihiazol-2-yl)-2,4- pyrtmidinediamine;
/V-(2-{[4-[(6-bromo-1 ,3-benzothiazol-2-yl)amino]-6-(4-morpholinylmethyl)-2- pyrimidinyl]amino}ethyl)-2-propenamide;
A/2-[(3S)-1 -acryloyf-3-pyrrolidSny[3- \/^(6-bromo-1 ,3-benzothiazol-2-yl)-6-(4- morpholinylmethyl)-2,4-pyrimidinediamine;
A/-(1 -acryloyl-4-piperidinyl)-4-(4-morpholinyimethyl)-W 1 ,33thiazolo[5l4-b]pyri pyridinediamine; N-[(3S)-1-acryloyl-3-pyrrolidinyl]-^
y!-2,6-pyridinediamine;
W-{1-acryloyi-3-piperidinyi)-4-(4-morphoiinylmethyl)-W 1 ,3]thiazolo
pyridinediamine;
A/ -(1 -acry!oyl-4-piperidinyl)-6-{[c/s-2,6^
[1 ,3]thiazolo[5,4-ti]pyridin-2-yi-2,4-pyriniidinediamine;
/V2-[(3S)-1-acryloy!-3-pyrrolidinyl]-6-^
[1 ,3]thiazolo[5,4-/)]pyndin-2-yt-2,4-pyrirnidinediamine;
A/-(2-{[4-{[c/s-2,6-dimethyi-4-morpholinyl]methyi}-6-{[1 ,3]thiazolo[5,4-0]pyridin-2-yiamino)- 2-pyrimidinyl3amino}ethyl)-2-propenamide;
A/-(1 -acryloyl-4-piperidinyl)-4-{[c/s-2,6^
jb]pyridin-2-yl-2,6-pyridinediarriirie;
W-[(3S)-1 -acryloyl-3-pyrrolidinyl]-4-{[c^
[l ^^hiazoioCS^-^pyridin^-yl^^-pyridinediainine;
/V-(1-acryloyl-3-piperidiny!)-4-{[c/s-2,6-^^
b]pyridin-2-yl-2,6-pyridinediamine;
A/-[<3S)-1-acryloyl-3-piperidinyl]-4-{[c/^
[1 ,3]thiazoiot5,4-0]pyridin-2-y[-216-pyridinediamine;
/V-[{3R)-1-acryloyl-3-piperidinyl]-4-{[c/s-2,6-dimeihy!-4-morpholinyi]meihyi}- ^
[1 ^lthiazo!oCS^-^pyridin^-yl^^-pyridinediamine;
W-{1 -[(2£)-4-(dimethylamino)-2-bute^
morpholinyiJmethylJ-A/'-II .SlthiazolotS^-^pyridfn^-yl^^-pyridinediamine;
/V-{(3S)-1-[(2£)-4-(dimethylamino)-2-butenoyl]-3-pyrrol!dinyl}-4-{tc s-2,6-dimeth morpholinytjmethylJ-A/'-II .SlthiazofofS^-^lpyridin^-yl^.e-pyridinediamine;
W-{1-[(2E)-4-(dimethylamino)-2-butenoy^
morpholinyl]methyi}- \/'-[1 ,3]thiazoto[5,4-&]pyridin-2-y[-2,6-pyr!dinediamine;
(2E)-4-(dimethy!amino)-A/-(2-{[4-(4-morp ^
ylamino)-2-pyrimidiny!]amino}ethyl)-2-butenamide;
A/2-{1 -[(2£)-4-(dimethy[amino)-2-buten^^
[1 ,3]thiazolo[5,4-/)3pyridin-2-yl-2,4-pyrimidinediamine;
W-{(3S)-1-[(2E)-4-(dimethylamino)-2-butenoyl]-3-pyrrolidfnyl}-4-(4-morpholinylmeih
[1 ,3]thiazolo[5,4-6]pyridin-2-yl-2,6-pyridinediamine;
W-{1-[(2£)-4-(dimethy!amino)-2-butenoyl]-4-piperidinyi}-4-(4-morpholinylmethyl)^
[1 ,33thiazo!o[514-0]pyrfdin-2-yi-216-pyridinediamine;
/V-{1 -[(2E)-4-(dimethy!amino)-24 rt^
[1 ,3]thiazolo[5,4-iE)]pyridin-2-yl-2,6-pyridinediamine; A -(1-acryloyl-4-piperidinyl)-4~(1-piperidinylrnethyl)-A/,-[1 ,3]thiazolo[5:4~0]pyridin-2-yi-2,6- pyridinediamine;
Λ/-(1 -acryloyl-4-piperidinyt)-A/'-[1 ,3]thiazolo[5l4-0]pyridin-2-yi-4-({[(1 S)-1 ,2,2- trimethy!propyS]amino}methyi)-2,6-pyridinediamine;
A -(1 -acryloyl-4-piperidinyi)-A , 1 ,3]thiazolo[5,4-0]pyridin-2-y!-4-({[(1 R)-1 l2J2- irimeihylpropy!]amino}meihy!)-2,6-pyridinediamine;
A/-(1 -acryioyl-4-piperidinyi)-4-[(2-methyl-1-pyrro[idinyl)methyl]-/\/'-[1 ,3]thiazolo[5,4- jb]pyridin-2-yl-2,6-pyridinediamine;
W-(1-acryloyl-4-p!peridinyl)-4-[(2,2-dimethyl-4-morpholinyl)methyl3-A/'-t1 ,3]thiazolo[5,4- kjpyridin-2-yl-2,6-pyridinediamirie;
A/-[(3S)-1 ~acryloyl-3-pyrrolidinyl]-4-(1-pipe^
2,6-pyridinediamine;
W-[{3S)-1-acryioyl-3-pyrrolidinyi]-4-t(2,2-dimeihyl-4-morphol!nyl)methyl]-A/'- [1 ,3]thsazolo[5,4-0]pyridin-2-yl-2,6-pyridinediamine;
W-[(3S)-1-acryloyl-3-pyrrolidinyl]-4-[(4-m^
j ]pyndin-2-y!-2,6-pyridinediamine;
^-[(SSJ-l-acryloyi-S-pyrrolidinyn-Ar-Il .^thiazoloiS^-blpyridin^-yl^ I R)-! ^^- trimethylpropyl3amino}meihy!)-2,6-pyridinediamine;
W-[(3S)-1 -acryloy!-S-pyrrolidinyll-W-tl ,3]thiazo!o[5,4-t>3pyridin-2-yl-4-({[(1 S)-1 ,2,2- trimethylpropyl]amino}methyl)-2,6-pyridinediamine;
W-(1 -acryloyl-4^iperidinyl)-4-[(dimethyiamino)methyl]- \/l-[1 ,3]thiazolo[5,4-/b]pyridin-2-yl-
2,6-pyridinediamine;
W-t(3S)-1-acryloyl-3-piperidinylH-(4-m
2,6-pyridinediamine;
V-[(3R)-1-acryloyl-3-piperidiny!]-4-(4-morphol^
2,6-pyridinediamine;
4-{[c/s-2,6-dimethyl-4-morpholinyl]methyl}-/V-[(3S)-1 -(3-methyl-2-butenoyl)-3-pyrroiidin
Af-[1 ,3]thiazolo[5,4-b]pyridin-2-yl-2,6-pyridinediamine;
W-{(3S)-1-[(2£)-2-butenoyl]-3-pyrrolrdinyl}-4-{[c/s-2,6-dimethyl-4-morp
[1 ,3]thiazo!o[5,4-i ]pyridin-2-yl-216-pyridinediamine;
4-{[c/ 2,6-dimeihyl-4-morpholinyl]methyl}-A/-{(3S)-1 -[(2E)-2-methyl-2-butenoyl]-3- pyrrolidinyi}-A/l-[1 ,3]ihiazolo[5,4-/)3pyridin-2-yl-2,6-pyridinediamine;
4-{[c s-216-dimethy!-4-morphoiinyl]methyl}- \/-t(3S)-1 -(2-methylacryloyl)-3-pyrrolid
[1 ,3]thiazolo[5>4-/b]pyridin-2-yl-2,6-pyridinediamine;
A/-{(3S)-1-[(2E)-4-(dimethylamino)-2-butenoy^
morpholinyl3meihyl}- /,-f1 l3]thiazo!o[5,4-£)]pyridin-2-yi-2,6-pyridinediamine;
4-{[c/s-2,6-dimethyl-4-morpholinyl]methyl}-A/-{(3S)-1 -[2-(4-morpholiny!methyl)acryloyl]-3- pyrroiidiny^-W-CI .SlthiazolotS^-bJpyridin^-yl-Z.e- yndinediarnine;
v"-(1 -acryloyl-3-azetidiny!)-4-(4-morpholinylm^
pyridinediamine;
A/-[(3S)-1-aciy!oyl-3-pyrrolidinyl]-4-[(dimethylam
2-yl-2,6-pyridinediamine;
W-[(3 ?)-1 -acrytoyl-3-pyrrolidinyl]-yV-I1 ,3]thiazoloI5,4-/)]pyridin-2-yl-4-({t<1 S)-1 ,2,2- trimethylpropyl]amino}methyl)-2,6-pyridinediarnine;
W2-[(3S)-1-aci7loyl-3-pyrrolidinyl]-^^
pyrimidinediamine;
A/-[(3S)-1-acry[oyl-3-pyrrolidinyl]-/\T-1 ,3-benzoihiazol-2-yl-4-(4-morphoiinylmeih pyridinediamine;
A/-[(3S)-1-acryloyl-3-pyrrolidinyl]-A/'-[6-(methyloxy)-1 ,3-benzothiazol-2-yl]-4-(4- morpholinylmethyl)-2,6-pyridinediamine;
^-[(SSJ-l-acryloyl-S-pyrroiidinyll-W-l ^-benzothiazol^-yl^-^KI SJ-l ^^- trimethylpropyl]amino}methyl)-2,6-pyridinediamine;
W-[(3S)-1 -acryloyl-3-pyrroiidinyl]-A/'-[6-(methyloxy)-1 ,3-benzothiazol-2-y!]-4-({[(1 S)-1 ,2,2- trimethylpropyl]amino}methyl)-2!6-pyridinediamine; and
W-[(3R)-1-acryloyl-3-pyrroiidinyi]-W-[1 ,3]thiazolo[5,4-0]pyridin-2-y!-4-({[(1 ,2,2- trimethylpropyl]amino}methyl)-2,6-pyridinediamine; and
salts thereof.
In another embodiment, the compound of the invention is selected from the group consisting of:
W-(2-{[4-[(methyloxy)methyl]-6-([1 ,3]thiazolo[5,4-f>]pyridin-2-ylamino)-2- pyrimidinyl]amino}ethyl)-2-propenamide;
A/-[2-({4-[(6-bromo-1 ,3-benzothiazol-2-yl)amino]-2-pyrimidiny[}amino)ethyl]-2- propenamide;
irans-4-{[4-(4-morpholinylmethyl)-6-([1 ,3]thiazolo[5,4-i)]pyndin-2-ylamino)-2- pyrimidinyl]amino}cyclohexyl methanesulfonate;
A/^il-acryloyl-S-pipendiny -e-timethylox Jmethy -W^EtSlthiazoloIS^-^pyridin^-yl^^- pyrimidinediamine;
A/^fl-acr loy!^-piperidin lJ-e-fimethyloxyJmethyil-A/^fl ^jthiazoloIS^-ijjpyridin^-yl^^- pyrimidinediamine;
irans-4-{E4-[{methyloxy)methyl]-6-([1 l33thiazoio[5,4-i>]pyridin-2-ylamino)-2- pyrimidinyl3amino}cyc!ohexyl methanesulfonate;
/V-(4-{E4-[(methyloxy)methyl]-6-(E1 l3]thiazolo[5l4-b]pyridjn-2-ylamino)-2- pyrimidinyl]amino}butyi)-2-propenamide; W-(3-{[4 (methyloxy)methyl]-6-([1 I3]thiazolo[5l4-b]pyridin-2-ylamino)-2- pyrimidinyi]amtno}propy!)-2-propenamide;
A/-(frans-2-{[4-[(methyloxy)methyl]-6-(i1 ,3]thiazolof5,4-0]pyndin-2-ylamino)-2- pyrimidinyl3amino}cyclohexyl)-2-propenamide;
W-(c/s-3-{[4 (methyloxy)methyl]-6-([1 ,3]thiazoio[5,4-b3pyridin-2-ylamino)-2^ pyrimidinyl]amino}cyclohexyl)-2-propenamide;
A -{irans-3-{[4-[{methyloxy)methyl]-6-([1 ,3]thiazolot5,4-0]pyridin-2-ylamino)-2- pyrimidinyl]amino}cyclohexyl)-2-propenarnide;
A/2-[(3S)-1-acry!oyl-3-pyrrolidinyl]-6-[(m^
2,4-pyrimidinediamine;
/V-{2-(4-acryioyl-1-piperazinyl)-6-[(methyloxy
)]pyridin-2-amine;
/V-(c s-2-{[4-[(methy!oxy)methyl]-6-([1 ,3]thiazolo[5,4-b3pyridin-2-ylamino)^ pynmidinyi]atnino}cyclohexyl)-2-propenatnide;
W-(4-{[4-[(methyloxy)methyl]-6-([1 ,3]thiazo!o[5,4-0]pyridin-2-ylamino)-2- pyrimidiny!]amino}cyclohexyl)-2-propenamide;
W-iirans^-i^- methylox Jmethy -e-it Slthiazolo^^-j J yridin^-ylamino)^- pyrimidinyl]amino}cyclohexyl)-2-propenamide;
W2-[(3R)-1-acryloyl-3-pyrrolidinyl3-6-[^
2,4-pyrimidinediamine;
W-{2-[(1-acryloyl-4-piperidinyl)oxy]-6-[(methyto
ij]pyridin-2-amine;
(2£)-4-(dimethylamino)-W-(2 [4-[(methyloxy)methyl]-6-([1 ,3]thiazolo[5,4-/}3pyndin-2- ylamino)-2-pyrimidinyl]amino}ethyl)-2-butenamide;
W^1-[(2E)-4-(dimethyiamino)-2-buteno^
[1 ,3]thiazolo[5,4-jb]pyridin-2-y!-2,4-pyrimidinediamine;
A/2-{1-[(2£)-4-(dimethylamino)-2-butenoyl]^
[1 ,3]thiazolo[5,4-j[)]pyridin-2-yl-2,4-pynrriidinediamine;
A -(2-{[4-(4-morpholiny!methyj)-6-([1 ,3]thtazolo[5,4-0]pyridin-2-ylamino)-2- pyrimidinyl3amino}ethyl)-2-propenainide;
A/-(ira/7s-2-{[4-(4-morpho]inyimethyl)-6-{[1 ,3]thiazolo[5,4-6]pyridin-2-ylamino pyrimidinyl]amino}cyc[ohexyi)-2-propenamtde;
A/2-(1 -acryloyi-3-piperidinyl)-6-(4-morpholiny^^
pyrimidinediamine;
Λ/~(ίΓans-4-{[4-(4-moφho!inylmethyl)-6-([1 3]ίhtazolot5,4-ib]pyridin-2-yjamin pyrimidinyl3amino}cyclohexyl)-2-propenamide; A/2-[(3R)-1-acryloyl-3-pyrrolidin^
yl-2,4-pyrimidinediamine;
A/-(c/s~4-{[4-(4-morpho!inylmethy!)-6-([1 !3]t^^
pyrimidinyl]amino}cyciohexyl)-2-propenamide;
A/2-(1-acryloyl-4-piperidinyl)-6-(4-morpholm^
pyrimidinediamine;
/V2-[(3S)-1 -acryloyl-3-pyrrolidinyl]-6-(4-m^
yl-2,4-pyrimidinediamine;
W-{c/'s-2-{[4-(4-morphoiinylmethyl)-6-([1 [3]thiazolo[5,4-63pyridin-2-ylamino)-2^ pyrimidiny!]amino}cyclohexyl)-2~propenamide;
A -(1-{6-t(6-bromo-1 ,3-benzothiazol-2-yl)amino]-4-[(2-methyi-1 H-imidazo!-1 -y!)methyl]-2- pyridinyl}-3-piperidiny[)-2-propenamide;
W-[2-(4-acryloyl-1-piperazinyi)-6-(4-morphoiinyimethyl)-4-pyrimidinyl][1 ,3]thiazolot5,4- b]pyridin-2-amine;
A/-[2-[(1-acryloyl-4-pipendinyl)oxy]-6-(4-morpho!inylmethyl)-4-pyrimidiny[][1 l3]thiazolo[5 i>]pyridin-2-amine;
W-(c/s-3-{[4-(4-morpholinylmethy!)-6-([1 ,3]
pyrimidiny[]amino}cyclohexyl)-2-propenamide;
A/-(ira/7s-3-{[4-(4-morpholinylmethyl)-6-([113]thiazolo[5,4-^]pyridin-2-ylamin
pyrimidinyl]amino}cyclohexyl)-2-propenamide;
A/2-[1-(chloroacetyl)-3-piperidiny!]-6^
yl^^-pyrimidinediamine;
W-[2-(2-acryioyitetrahydro-1 (2H)-pyridazinyl)-6-(4-morpholinylmethyl)-4- pyrimidiny^l ^lthiazoioiS^-feJpyridin^-amine;
/V2-(1-{[(4-fluorophenyl)oxy]acetyl}-3-piperidinyi)-6-(4-morpholinylmethyl)-/V'i- tl .SjthiazolofS^-^pyridin^-yl^^-pyrimidinediamine;
2-chloro-W-(2-{[4-(4-morpholinylmeihyl)-6-([1 ,3]thiazolo[5^-6]pyridin-2-ylamino)-2- pyrimidjnyl]amino}ethyl)acetamide;
(2£)-4^^-trifiuoro-A/-(2-{[4-(4-morpholinylmethyl)-6-([1 ,33thiazolo[5l4- )]pyndin-2- ylamino)-2-pyrimidinyi]amino}eihyl)-2-buienamide;
2^{44luorophenyi)oxy]-W-(2-{[4-(4-morpholinylmethyl)-6-([1 3thiazolo[5,4-i)]pyndin-2- ylamino)-2-pyrimidinyi]amino}ethy!)acetamide;
W-{1 -[4-(4-morpholinyimethyl)-6-([1 ,3]thiazojo[5,4-03pyridin-2-y!amino)-2-pyridin piperidinyl}-2-propenamide;
W-(c/s-3-{[4-(4-morphoiinylmethyl)-6-^^
pyridinyl]amino}cyclohexyl)-2-propenamide; A/-(fra/?s-3-{[4-(4-morpholinylmethyl)-6-([1 ,3]thiazolo[5,4-^3pyridin-2-y!ami pyridinyl]amino}cyclohexyt)-2-propenamide;
/-(2-{[4-(4-morpholinylmethyl)-6-([1 ,3]thiazo[o[5,4-0]pyndin-2-ylamino)-2- pyridinyl]amino}ethyl)-2-propenamide;
A/-c/s-2-{[4-(4-morpholinylmethyl)-6-([1 ,3]thiazolo[5,4-0]pyrSdin-2-ylamino)-2- pyridinyl]amino}cyclohexyl)-2-propenamide;
A/-{/ra^s-2-{[4-(4-morphoiinylmethyi)-6-([1 ,3]thiazo[o[5^-0]pyridin-2-ylamino)-2- pyridinyl]amino}cyclohexyl)-2-prop6namide;
6-(4-morpholinylmethyl)-W*-[1 ,3]thiazolo[5,4-6]pyridin-2-yl-W2-{1-[(2£)-4,4,4 butenoyl]-3-piperidinyl}-2,4-pyrimidinediamine;
A/-methyl-W-(2-{[4-{4-morpholinylm^
pyrimidinyi]amino}ethyi)-2-propenamide;
A/-{3-[4-(4-morpho[inylmethyl)-6-(E1 ,3]thiazolo[5l4-6]pyridin-2-ylamino)-2- pyrimidiny!]pheny!}-2-propenamide;
W-[4-{4-morpholinylrnethyl)-6-([1 ,3]thiazoio[5,4-03pyridin-2-ylamino)-2-pyridinyl]-2- propenamide;
AT-[4-(4-morpholinylmethyl)-6-^
pyrirriidinylidene]-2-propenohydrazide;
A/ -[(3S)-1 -acryioyl-3-pyrro!idinyl]-Ar'-(6-bromO"1 ,3-benzothiazol-2-yl)-2,4- pyrimidinediamine;
W-(2-{[4-[(6-bromo-1 J3-benzothiazol-2-yl)amino]-6-(4-morpholinylmethyl)-2- pyrimidinyl]amino}ethyl)-2-propenamide;
/V2-t(3S)-1-acryloyl-3-pyrro!idinyl]- \r'-(6-bromo-1 [3-benzothiazol-2-yl)-6-(4- morpholinylmethyl)-2^pyrimidinediamine;
A/-(1 -acryloyl-4~piperidinyl)-4-(4-morpholinylm^
pyridinediamine;
A/-[(3$)-1 -acryloyl-3-pyrrolidinyl]-4-(4-mor^
yl-2,6-pyridtnediamine;
W-(1-acryloyl-3-piperidinyl)-4-(4-morpholinylm
pyridinediamine;
A/2-(1-acryloyl-4-pipe diny!)-6-{[c/'s-2,6^
[1 ,3]thiazolo[5,4-jfj]pyridin-2-yl-2,4-pyrimidinediamine;
A/2-[(3S)-1 -acryioyl-3-pyrrolidinyl]-6-{[c/s-2,6-dimethyl"4-morpho[iny!]meth
[1 ,33thiazolo[5,4-i)]pyridin-2-y[-2,4-pynmidinediamine;
A/-(2-{[4-{[c/s-2,6-dimethyl-4-morphoiinyi]methyl}-6-([1 >3]thiazoio[5l4-b]pyrid
2-pyrimidiny[3amino}ethyl)-2-propenamide; W-(1 -aci7loyl-4-piperidinyl)-4-{[c/s-2,6-dimethyl-4-morpho!inyl]m
6jpyridin-2-yl-2,6-pyridinediamine;
W-[{3S)-1 -acryloyl-3-pyrrolidinyl]-4-{[c^
t1 ,3]thiazolo[5,4-0]pyridin-2-yl-2,6-pyridinediamine;
A/-(1 -acry[oyi-3-piperidinyl)-4-{[c s-2,6-dimet^^
6]pyridin-2-yS-2,6-pyridinediamine;
Λ/-[(3S)-1-acryloyl-3-p!peπdίnyl]-4-{[c/s--2l6-dimeίhyl-4-morphoίinyl3methyί}-Λ/,- [1 ,3]thiazolot5,4-0]pyridin-2-yl-2,6-pyridinediamine;
Λ/-[(3R)-1-acΓy[oyl-3-p!peΓidinyl]-4 [c's-2,6-dimeίhyl-4-morpho!inyl3methyl}-Λ/,- [1 ,3]thiazolo[5,4-Jb]pyndin-2-yl-2,6-pyridinediamine;
W-{1-[(2£)-4-(dimethylamino)-2-butenoyl]-4-piperidinyl}-4-{[c s-2,6-dimeth
morphoiinyl]methyl}-A^[1 ,33thta^^
W-{(3S)-1 -[(2£)-4-(dimethylamino)-2-buienoyl3-3-pyrrolidinyi}-4-{[c/'s-2I6-di
morpholinyllmethy^-W-i SJthiazolofS^-^Jpyridin^-yl^.e^yridinediamine;
Λ/-{1 -[(2E)-4-(dimethylamino)-2-buienoyl]^
morpho!inyljmethyl^/V-II ^Jthiazo!oES^-bjpyridin^-yl^^-pyridtnediamine;
(2E)-4-(dimethylamino)-W-(2-{[4-(4-mo
ylamino)-2-pyrimidinyl]amino}ethyl)-2-butenamide;
A/2-{1 -[(2E)-4-(dimethy!amino)-2-but^^
[1 ,3]ihiazolo[5,4-ii]pyridin-2-yl-2,4-pynmidinediamine;
A/-{(3S)-1 -[(2£)-4-(dimethySamino)-2-butenoyl]-3^yrroiidinyl}-4-(4-morpholinylm
[1 ,3]ihiazolo[5,4-/)]pyridin-2-y!-2,6-pyridinediamine;
A/-{1 -[(2E)-4-(dimethylamino)-2-butenoyl]-4-piperidinyl}-4-(4-morpholinylme
[1 ,3]ihtazolo[5,4-/b]pyridin-2-yi-2,6-pyridinediamine;
W-{1 -E{2E)-4-(dimethyiamino)-2-butenoyl]-3-piperidiny[}-4-(4-morphoiinyS
[1 ,3]thiazolo[5,4-jb]pyridin-2-yl-2,6-pyridinediamtne;
Λ/-{1 -acryloyl-4-piperidinyl)-4-(1 -piperidinylmethyi)-/V-[1 ,33thiazolo[5,4-0]pyridin-2-yl-2,6- pyridinediamine;
W-(1 -acryloyl-4-piperidinyl)-W-[1 ,3]thiazoloE5,4-/5]pyridin-2-y!- -({[(1 S)-1 ,2,2- trimethylpropyl]amino}methyl)-2,6-pyridinediamine;
W-(1-acryloyl-4-piperidinyl)-^[1 ,3]tN^^
trimeihyipropyl]amino}methyl)-2,6-pyridinediamine;
A/-(1-acryloyl-4-piperidinyi)-4-[(2-methyf-1 -pyrrolidinyl)methylJ-A/'-[1 ,3]t iazolo[5,4- 0]pyridin-2-yl-2,6-pyridinediamine;
/V-(1 -acryloyi-4-piperidinyl)-4-[(2,2-dim^
£]pyridin-2-yl-2,6-pyridinediamine; A -E<3S)-1-aci^ioyl-3-pyrroifdinyI]-4-(1 -piperidinylmethyl)- '-I1.SJthiazolotS^-j Jp ridin^-yl- 2,6-pyridinediamine;
Λ/-E{3S)-1-acryloyl-3-pyΓΓOlidiny!]-4-[(2l2-dimeth ]-4-morpho[inyl)rnethy[3-Λ/,- [I .SJthiazolotS^-fclpyridin^-yl^.e-pyridinediamine;
/V-[(3S)-1-acryioyl-3-pyrrolidinyl]-4-[(4-methy
b3pyridin-2-yi-2,6-pyridinediamine;
W-[(3S)-1-acryioyl-3-pyrrolidinyl]-AH1 ,3^
trimethylpropyi]amino}methyl)-2,6-pyridinediamine;
^-[(SSJ-l-acryioyl-S-pyrrolidinyll-W-I SlihiazoioiS^-^pyridin^-yl^-a I SJ-l ^^- trimethylpropyi]amino}methyl)-2,6-pyridinediamine;
A/-(1 -acryloyl-4-piperidinyl)-4-[(dimethy!amino)methy^
2,6-pyridinediamine;
A/-[(3S)-1-acryioyl-3-piperidiny!]-4-(4-morpholinylmei^^^
2,6-pyridinediamine; and
N-[(3R)-1-acryloy!-3-piperidiny!]-4-(4-mo
2,6-pyridinediamine; and
salts thereof.
In another embodiment, the compound of the invention is selected from the group consisting of:
/-(2-{[4-[{methyloxy)methyl]-6-([1 ,33thiazoio[5,4-b]pyridin-2-yiamino)-2- pyrimidinyljamino}ethyl)-2-propenamide;
A/-[2-({4-[(6-bromo-1 ,3-benzothiazol-2-yl)amino]-2-pyrimidinyl}amino)ethyl]-2- propenamide;
rrans-4-{[4-(4-morpholinylmethyi)-6-([1 ,3]thiazo!o[5l4-i)lpyridin-2-ylamino)-2- pyrimidinyljaminojcyclohexy! methanesulfonate;
W^il -acryloyl-S-piperidiny -e-iimethyloxyJmethyll-W^CI .SJthiazolotS^-i lpyndin^-yl^^- pyrimidinediamine;
^-(l -acryloyl^-pipendiny -G-IfmethyloxyJmethylJ-A ^-tl .SJthiazolotS^-jblpyridin^-yl^^- pyrimidinediamine;
ira/is^-t^-ffmethyloxyjmethylj-e^fl ^jthiazolots^-jbjpyridin^-ylamino)^- pyrimidinyl]amino}cyclohexyl methanesulfonate;
A/-(4-{[4-[(methyloxy)methyl]-6-([1 ,3]thiazoio[5l4-j ]pyridin-2-ylamino)-2- pyrimidinyl]amino}butyl)-2-propenamide;
W-(3-{[4-[(methyloxy)methyl]-6-([1 ,33thiazoio[5,4-b]pyndin-2-ylamino)-2- pyrimidinyl]amino}propyl)-2-propenamide; W-(ira 7s-2-{[4-[(meihyloxy)methyl3-6-([1 ,3]thiazoio[5,4-b]pyridin-2-ylaiTiino)-2- pyrimidinyl]amino}cyclohexyl)-2-propenamide;
W-(c/s-3-{[4-[(methyloxy)methyl]-6-(E1 ,3]thiazolo[5,4-/33pyridin-2-yla
pyrimidinyl]amino}cyclohexyl)-2-propenamide;
W-(ira ?s-3-{[4-[(methyloxy)methy[3-6-([113]thiazolo[5l4-0]pyridin-2-ylannino)-2- pyrimidinyl]amino}cyclohexyl)-2-propenamide;
A/2-[(3S)-1-acryloyl-3-pyrrolidinyl]-6-[(m^
2,4-pyrimidinediamine;
A/-{2-(4-acryloyl-1 -piperazinyl)-6-[(methyloxy)methyl]-4-pyrimidiny!}[1 ,33th!azolo[5,4- i>]pyridin-2-amine;
/V~(c/s-2-{[4-[(methy!oxy)met^
pyrimidinyl3amino}cyclohexyl)-2-propenamide;
A/-(4-{[4-[(methyloxy)methyl]-6-([1 ,3]thiazoio[5,4-b3pyridin-2-ylamino)-2- pyrimidiny1]amino}cyclohexyl)-2-propenamide;
A/-(irans-4-{[4-[(methyloxy)methy[j-6-([1 ,3]thiazolo[5,4-j ]pyridin-2-ylamino)-2- pyrimidinyi]amino}cyclohexyl)-2-propenamide;
W (3R)-1-acryioyl-3-pyrroiidinyl]-6-[(m^
2,4-pyrimidinediamine;
W-{2-[(1-acryloyi-4-piperidinyl)oxy]-6-[(methyloxy)methyl]-4-pynmidinyl}[1 ,3]thiazolo[5,4^ /b3pyridin-2-amine;
(2E)-4-(dimethylamino)-A/-(2-{[4-[(methyloxy)methyl]-6-([1 ,3]thiazolo[5,4-b]pyridin-2- ylamino)-2-pyrimidinyl]amino}ethyl)-2-butenamide;
W^1-[(2£)-4-(dimethy]amino)-2-butenoy^^
[1 ,3]thiazoioE5,4-j ]pyridin-2-yl-2,4-pynmidinediamine;
W2-{1-[(2£)-4-(dimethy!amino)-2-buten^
[1 ,3]thiazofo[5,4-£)]pyndin-2-yl-2,4-pynmidinediamine;
A/-(2-{[4-(4-morpholinylmethyl)-6-([1 ,3]thiazolo[5[4- ]pyndin-2-ylamino)-2- pyrimidinyi]amino}ethyl)-2-propenamide;
A/-(ira/7s-2-{[4-(4-morpholinylmeihyl)-6-([1 ,3]thiazo!o[5,4-d]pyridin-2-ylarriino)-2- pyrSmidinyl3amino}cyclohexyl)-2-propenamide;
/V2-(1-acryloyl-3-piperidinyl)-6-(4-morpholm^
pyrimidinediamine;
W-(irans-4-{[4-(4-morpholinylmethyl)-6-{[1 ,3]ihiazo[o[5,4-/b]pyridin-2-ylamin
py midinyl]amino}cyciohexyl)-2-propenamide;
W (3R)-1-acryloyl-3-pyrrolidiny!]-6-(4-mor^
yi-2,4-pyrimidinediamine; W-(c/s-4-{[4-(4-morpho!inylmethyl)-6-([1 ,33thiazo!o[5,4-b]pyridin-2-ylamin
pyrimidinyl]amino}cyc!ohexyl)-2-propenamide;
N2-(1-acryloyj-4-piperidinyl)-6-(4-morpholiny!methyi)-W4-[1 ,3]ihiazolo[5,4-b]pyri pyrimidinediamine;
A/2-[{3S)-1 -acryloyl-3^yrrolidinyl]-6-(4-morpho^
yl-2,4-pyrimidinediamine;
N-(c s-2-{[4-{4-morphoiinylmethyl)-6-([1 ,3]thiazolo[5,4- 5]pyridin-2-ylami
pyrimidinyl]amino}cyc!ohexyi)-2-propenamide;
W-(1-{6-[(6-bromo^3-benzothiazoi-2-yl)amino]^
pyridinyt}-3-piperidinyl)-2-propenamide;
A/-[2-(4-acry!oyl-1-piperazinyl)-6-(4-morpho!inylmethy!)-4-pyrimidinyl][113]thiazojo[5 6]pyridin-2-amine;
W-[2-[(1-acryloyl-4-piperidinyl)oxy]-6-(4-morpholinyimethyi)-4-pyrimidinyl][1 ,3]thi b]pyridin-2-amine;
A/-(c/≤-3-{[4-(4-morpholiny[methyl)-6-([1 ,3]thiazolo[5,4-b]pyridin-2-yjamino)-2- pyrimidiny!]amino}cyciohexyl)-2-propenannide;
W-(fra/?s-3-{[4-(4-morpholinyjmethyl)^
pyrimidiny!]amino}cyclohexyl)-2~propenamide;
A/2-[1 -(ch!oroacety!)-3-piperidinyl]-6-(4-^
yl^^-pyrimidinediamine;
W-[2-(2-acryloyltetrahydro-1 (2H)-pyridazinyl)-6-(4-rriorphoiinyinnethyl)-4- pyrimidiny!][1 ,3]thiazolo[5,4-/t)]pyridin-2-amine;
W2-(1-{[(4-fiuorophenyl)oxy]acetyl}-3-piperidinyf)-6-(4-morphoiinylmethyl)-/\Ti- [1 ,3]ihiazolo[5,4-ij]pyridin-2-yl-2,4-pyrimidinediamine;
2-chloro-A/-(2-{[4-(4-morpholinylmethy^
pyrimidiny!]amino}et yl)acetamide;
(2E)-4,4^-trifluoro-W-(2-{[4-(4-morpholinylmethyl)-6-([1 l3]thiazolo[5,4-03pyridin-2- yjamino)-2-pyrimidinyl]amino}ethyl)-2-butenamide;
2-[(4-fiuorophenyl)oxy]-A/-(2-{[4-{4-morpholinylmethyl)-6-([1 ,3]ihiazolo[514-/b]pyridin-2- y!amino)-2-pyrimidlnyl]amino}ethyl)acetamide;
A/-{1-[4-(4-morpholsnylmethyl)-6-([1 ,3)tM^
piperidinyl}-2-propenamide;
A/-(c/s-3-{[4-(4-morpholinylmethyi)-6-([1 ,3]thiazolo[5,4-/)]pyridin-2-ylamino)-2- pyridinyl]amino}cyclohexyl)-2-propenamide;
A/-(irans-3-{[4-(4-morpholinylmethyl)-6-([1 ^jthiazolotS^-jbJpyridin^-yiamino)^- pyridinyi]amino}cyciohexyl)-2-propenamide; A -(2-{[4-(4-morphoiinylmethyl)-6-{[1 ,3]thiazolo[5<4-6]pyridin-2-ylamino)-2- pyridinyl]amino}ethyl)-2-propenamide;
W-c/s-2-{[4-(4-morpholinylmethyl)-6-([1 ^^
pyridinyl]amino}cyclohexy!)-2-propenamide;
A/-(fra/is-2-{[4-(4-morpholinylmethyl)-6-([1 ,3]thiazojo[5,4-0]pyridin-2-ylam
pyridinyi]amino}cyclohexy!)-2-propenamide;
6-(4-morpho[inylmethyl)-Ar4l .3]t^
butenoyf]-3-piperidinyl}-2,4-pyrimidinedtamine;
A/-methyl-W-(2-{[4-(4-morpho1inyfmeth^
pynmidinyi]amino}eihyl)-2-propenamide; and
AZ-iS-^^-morpholinyimethy -e-iCl ^lthiazolotS^-^pyridin^-ylamino)^- pyrimidiny!]phenyl}-2-propenamide; and
salts thereof. In another embodiment, the compound of the invention is selected from the group consisting of:
A -c/s-2-{[4-(4-morpholinylmethyl)-6-([1 ,3]thiazolo[5,4-b]pyridin-2-ylamino)-2- pyridinyl]amino}cyclohexyl)-2-propenamide;
A/2-[(3S)-1 -acryloyl-3-pyrrolidinyl]-6-(4-m^
y!-2,4-pyrimidinediamine;
W2-(1-acry!oyl-4-piperidinyl)-6-(4-morph
pyrimidinediamine;
W-(c/s-4-{[4-(4-morphoiinylmethyl)-6-([1 ,3]thiazo[o[5,4-i ]pyridin-2-yiamino)-2-- pyrimidinyl]amino}cyclohexyl)-2-propenamide;
Λ/2-(1 -acry!oyl-3-piperidinyl)-6-(4-mo hol!nylmethyl)-Ay4-t1 ,33thiazolot5,4- )]pyridin-2-yl-2,4- pyrimidinediamine; and
A/-(c s-3-{[4-(4-morpholinylmethy!)-6-([1 ,3]thiazolo[5!4-j ]pyridin-2-ylamino)-2- pyridinyi]amino}cyclohexyl)-2-propenamide; and
salts thereof.
In another embodiment, the compound of the invention is selected from:
A/-[(3S)-1-acryloyl-3-pyrrolidinyl]-4-{[c/s-2!6-dimethyl-4-morpholinyl]methyl}-A - [1 ,3]thiazolo[5,4-0]pyridin-2-yl-2,6-pyridinediamine; and
A/-[(3S)-1-acryloyl-3-pyrrolidinyl]-/V-[1 ,3]thiazoio[5,4-i>]pyndin-2-yl-4-({[(1 S)-1 ,2,2- trimethylpropyl]amino}methyS)-2,6-pyridinediamine; and
salts thereof. In another embodiment, the compound of the invention is:
Λ/-[(3S)-1-acr jo l·3-p rroiidinyl]-4-{[c s-2,6-dimethyl-4-morphol!nyl]methyl}-Λ/,- [1 ,3]thiazolo[5,4-^yridin-2-yl-2,6-pyridinediamine; or
a salt thereof.
In another embodiment, the compound of the invention is:
V-[(3S)-1-acry!oyi-3-pyrro!idinyl]-W-[1 ,3]thiazolo[5,4-6]pyridin-2-yl-4-({[(1 S)-1 ,2,2- trimethyipropyl]amino}methyl)-2,6-pyridinediamine; or
a salt thereof.
In another embodiment, the compound of the invention is:
/V-[1-acryloyl-3-pyrrolidinyl]-/V,-[1 l3]thiazo!o[5,4-03pyridin-2-yi-4-({[1 ,2,2- trimethylpropyi]amino}methyl)-2,6-pyridinediamine; or
a salt thereof.
In another embodiment, the compound of the invention is:
A/-t(3S)-1-acryloyl-3-pyrrolidinyl3-/V-[1 ,3]thiazolo[5,4-0)pyridin-2-yl-4-({[(1 S)-1 ,2,2- trimethylpropyi]amino}methyl)-2,6-pyridinediamine. In a further embodiment, the compound of the invention is:
A/-[(3S)-1-acrySoyi-3-pyrrolidinyi]-A/,-[1 ,3]thiazoio[5,4-63pyridin-2-yl-4-({[(1 S)-1 ,2,2- trimethylpropyl]amino}methyl)-2,6-pyridinediamine difumarate.
In one embodiment, the compound of formula (I) or a salt thereof is selected from the group consisting of:
/^-[(SSJ-l-acrySoyl-S-pyrroSidinyll^-itc/s^.e-dimethyl^-morpholinyljmethylJ-W-tS- (methyloxy)[1 ,3]thiazolo[5,4- ]pyridin-2-yl]-2,6-pyridinediamine;
A/-E(3S)-1-acryloyl-3-pyrrolidinyi]-A/,-i5-(methyloxy)[1 ,3]thiazoSo[5,4-to]pyridin-2-yl]-4-({[(1 S)- 1 ,2,2-trimethylpropyl]amino}methyl)-2,6-pyridinediamine;
W-ttasj-l-acryloyl-S-pyrrolidinyH-tidimethylaminoJmethyll-W-fS- (methyloxy)[1 ,3]thiazolo[5,4-0]pyridin-2-yl]-2,6-pyridinediamine;
V-[(3S)-1-acryloyl-3-pyrrolidinyi]-4-[(dimethylamino)methyi]- v1-[6-(methyloxy)-1 ,3- benzothiazol-2-yl]-2,6-pyridinediamine; and
A/-[(3S)-1-acryloyS-3-pyrroiidinyl3-A/'-1 ,3-benzothiazol-2-yl-4-[(dimethylamino)methyl]-2,6- pyridinediamine; and
salts thereof. Terms and Definitions
"AlkyI" refers to a saturated hydrocarbon chain having the specified number of member atoms. C1-6alkyl refers to an alky! group having from 1 to 6 member atoms, for example 1 to 4 member atoms. AlkyI groups may be optionally substituted with one or more substituents as defined herein. AlkyI groups may be straight or branched. Representative branched alkyl groups have one, two, or three branches. AlkyI includes methyl, ethyl, propyl (n-propyl and isopropyl), butyl (n-butyl, isobutyl, and t-butyl), pentyl (n-pentyl, isopentyl, and neopentyl), and hexyl. Alkyl groups may also be part of other groups, for example alkoxy.
"Alkynyl" refers to a hydrocarbon chain having the specified number of member atoms and at least one triple bond. For example, C2.6alkynyl refers to an alkynyl group having from 2 to 6 member atoms, for example 2 to 4 member atoms. Alkynyl groups may be straight or branched. Alkynyl includes ethynyl, 1-propynyl, 1 -butynyl, 2-butynyl, 1- pentynyi, 2-pentynyl, 3-pentynyl, 1-hexynyl, 2-hexynyi and 3-hexynyl.
"Cycloalkyl" refers to a saturated hydrocarbon ring having the specified number of member atoms. Cycloalkyl groups are monocyclic ring systems. For example, C3. ecycloalkyl refers to a cycloalkyl group having from 3 to 6 member atoms. Cycloalkyl groups may be optionally substituted with one or more substituents as defined herein. Cycloalkyl includes cyc!opropyl, cyc!obutyl, cyclopentyi, and cyc!ohexyi. In one embodiment, the cycloalkyl groups herein are cyclohexyl.
"Enantiomerically enriched" refers to products whose enantiomeric excess is greater than zero, For example, enantiomerically enriched refers to products whose enantiomeric excess is greater than 50% ee, greater than 75% ee, and greater than 90% ee.
"Enantiomeric excess" or "ee" is the excess of one enantiomer over the other expressed as a percentage. As a result, since both enantiomers are present in equal amounts in a racemic mixture, the enantiomeric excess is zero (0% ee). However, if one enantiomer was enriched such that it constitutes 95% of the product, then the enantiomeric excess would be 90% ee (the amount of the enriched enantiomer, 95%, minus the amount of the other enantiomer, 5%). "Enantiomerically pure" refers to products whose enantiomeric excess is 99% ee or greater. "Half-life" (or "half-lives") refers to the time required for half of a quantity of a substance to be converted to another chemically distinct species in vitro or in vivo.
"Halo" refers to the halogen radical fluoro, chloro, bromo, or iodo.
"Heteroaryl", unless otherwise defined, refers to an aromatic ring containing from 1 to 3 heteroatoms as member atoms in the ring or rings. Heteroaryl groups containing more than one heteroatom may contain different heteroatoms. Heteroaryl groups may be optionally substituted with one or more substituents if so defined herein. The heteroaryl groups herein are monocyclic ring systems or are fused bicyclic ring systems. Monocyclic heteroaryl rings have 5 or 6 member atoms. Bicyclic heteroaryl rings have 9 or 10 member atoms. Monocyclic heteroaryl includes pyrrolyl, furany!, pyrazolyl, imidazo!yl, oxazolyl, isoxazolyl, oxadiazoiyl, triazolyl, pyridinyl, pyrimidinyl, pyridazinyl and pyrazinyi. Bicyclic heteroaryl includes indolyl, isoindolyl, indolizinyl, indazolyl, benzimidazoly!, pyrrolopyridinyl, quinoiinyl, isoquinoiinyl, quinoxalinyl, quinazolinyl and naphthridinyi. In one embodiment, the heteroaryl groups herein are monocyclic ring systems having 5 member atoms, for example pyrrolyl, pyrazolyl or imidazolyl.
"Heteroatom" refers to a nitrogen, sulphur, or oxygen atom.
"Heterocyclyl", unless otherwise defined, refers to a saturated or unsaturated ring containing 1 or 2 heteroatoms as member atoms in the ring. However, heterocyclyl rings are not aromatic. In certain embodiments, heterocyclyl is saturated. In other embodiments, heterocyclyl is unsaturated but not aromatic. Heterocyclyl groups containing more than one heteroatom may contain different heteroatoms. The heterocyclyl groups herein are monocyclic ring systems having 4, 5 or 6 member atoms. Heterocyclyl groups may be optionally substituted with one or more substituents as defined herein. Heterocyclyl includes azetidinyl, pyrrolidinyl, pyrazo!idinyl, imidazolinyl, oxazoiidiny!, isoxazolidinyl, piperidinyl, piperazinyl and morphoiinyl. In one embodiment, the heterocyclyl groups herein are monocyclic ring systems having 5 or 6 member atoms, for example pyrrolidinyl, piperidinyl, piperazinyl, tetrahydro-1 {2H)-pyridazinyl and morphoiinyl.
"Member atoms" refers to the atom or atoms that form a chain or ring. Where more than one member atom is present in a chain and within a ring, each member atom is covalently bound to an adjacent member atom in the chain or ring. Atoms that make up a substituent group on a chain or ring are not member atoms in the chain or ring. "Optionally substituted" indicates that a group, such as heteroaryl, may be unsubstituted or substituted with one or more substituents as defined herein. "Substituted" in reference to a group indicates that a hydrogen atom attached to a member atom within a group is replaced. It should be understood that the term "substituted" includes the implicit provision that such substitution be in accordance with the permitted valence of the substituted atom and the substituent and that the substitution results in a stable compound (i.e. one that does not spontaneously undergo transformation such as by rearrangement, cyclization, or elimination). In certain embodiments, a single atom may be substituted with more than one substituent as long as such substitution is in accordance with the permitted valence of the atom. Suitable substituents are defined herein for each substituted or optionally substituted group. "Pharmaceutically acceptable" refers to those compounds, materials, compositions, and dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, or other problem or complication, commensurate with a reasonable benefit/risk ratio.
As used herein the symbols and conventions used in these processes, schemes and examples are consistent with those used in the contemporary scientific literature, for example, the Journal of the American Chemical Society or the Journal of Biological Chemistry. Unless otherwise noted, all starting materials were obtained from commercial suppliers and used without further purification. Specifically, the following abbreviations may be used in the examples and throughout the specification:
DCM dichloromethane
DMF dimethylformamide
DMSO dimethylsulphoxide
Et ethyl
HATU O-CZ-azabenzotriazol-l-y -W. A/'.A/'-tetramethyiuronium
hexafiuorophosphate
HPLC high performance liquid chromatography
I PA /so-propanoi
LCMS liquid chromatography-mass spectrometry
DAP mass-directed autopreparative HPLC Me methyl
min minutes
mg milligrams
mL mi!li!itres
mM mi!limolar
mmol mi!limo!es
m/z mass/charge ratio
N P W-methyl-2-pyrro!idone
NMR nuclear magnetic resonance
Pr n-propyl
Rt retention time
TFA trifluoroacetic acid
THF tetrahydrofuran
UPLC ultra performance liquid chromatography
UV ultraviolet
All references to brine are to a saturated aqueous solution of NaCI.
Included within the scope of the "compounds of the invention" are al! solvates (including hydrates), complexes, polymorphs, prodrugs, radioiabel!ed derivatives, stereoisomers and optical isomers of the compounds of formula (I) and salts thereof.
The compounds of the invention may exist in solid or liquid form. In the solid state, the compounds of the invention may exist in crystalline or noncrystalline form, or as a mixture thereof. For compounds of the invention that are in crystalline form, the skilled artisan will appreciate that pharmaceutically acceptable solvates may be formed wherein solvent molecules are incorporated into the crystalline lattice during crystallization. Solvates may involve nonaqueous solvents such as ethanol, isopropanoi, DMSO, acetic acid, ethano!amine, and EtOAc, or they may involve water as the solvent that is incorporated into the crystalline lattice. Solvates wherein water is the solvent that is incorporated into the crystalline lattice are typically referred to as "hydrates." Hydrates include stoichiometric hydrates as well as compositions containing variable amounts of water. The invention includes ail such solvates. In one embodiment, the compound of the invention is /V-[{3S)-1-acryloyl-3-pyrrolidinyl]-/V-[1 ,3]thiazolo[5,4-&]pyridin-2-yl-4-({[(1 S)- 1 ,2,2-trimethyipropyl]amino}methy[)-2,6-pyridinediamine difumarate monohydrate. The skilied artisan will further appreciate that certain compounds of the invention that exist in crystalline form, including the various solvates thereof, may exhibit poiymorphism (i.e. the capacity to occur in different crystalline structures). These different crystalline forms are typically known as "polymorphs". The invention includes all such polymorphs. Polymorphs have the same chemical composition but differ in packing, geometrical arrangement, and other descriptive properties of the crystalline solid state. Polymorphs, therefore, may have different physical properties such as shape, density, hardness, deformabi!ity, stability, and dissolution properties. Polymorphs typically exhibit different melting points, IR spectra, and X-ray powder diffraction patterns, which may be used for identification. The skilled artisan will appreciate that different polymorphs may be produced, for example, by changing or adjusting the reaction conditions or reagents, used in making the compound. For example, changes in temperature, pressure, or solvent may result in polymorphs. In addition, one polymorph may spontaneously convert to another polymorph under certain conditions.
In one embodiment, the present invention provides W-[(3S)-1-acryloyl-3-pyrrolidinyl]-W- [1 ,3]thiazoio[5,4-b]pyridin-2-yl-4-({[(1 S)-1 ,2,2-trimethylpropyi]amino}methyl}-2,6- pyridinediamine or a pharmaceutically acceptable salt thereof in crystalline form. In another embodiment, the present invention provides A/-[(3S)-1-acryloyl-3-pyrrolidinyl]- /V-[1 ,3]thiazolo[5,4-/b]pyridin-2-yl-4-({[(1 S)-1 ,2,2-trimethyipropyl]amino}methyi)-2,6- pyridinediamine difumarate monohydrate in crystalline form.
In another embodiment, the present invention provides a polymorph of /v/-[(3S)-1~acryloyl- 3-pyrro!idinyl]- V-[ ,3]thiazolo[5,4-6]pyridin-2-y!-4-({[(1 S)-1 ,2,2- trimethylpropyl]amino}methyl)-2,6-pyridinediamine difumarate monohydrate characterised in that it provides an XRPD pattern comprising peaks (°2Θ) at about 4.5, about 7.5 and/or about 8.7. In another embodiment, the present invention provides a polymorph of W-[(3S)-1-acryloyl- 3-pyrrolidinyl]-W-[1 ,3]thiazolo[5,4-i)]pyridm-2-yi-4-({t{1 S)-1 ,2,2- trimethylpropyl]amino}methyi)-2,6-pyridinediamine difumarate monohydrate characterised in that it provides an XRPD pattern comprising peaks substantially as set out in Table 1. In a further embodiment, the present invention provides a polymorph of /V-[(3S)-1 -acryioyi- S-pyrrolidinylj-W-tL^thiazolors^-jbjpyhdin^-yl^-^I S)-! ^^- trimethy!propyi3amino}methyi)-2,6-pyridinediarriine difumarate monohydrate characterised in that it provides an XRPD pattern substantialiy in accordance with Figure 1.
When it is indicated herein that there is a peak in an XRPD pattern at a given vaiue, it is typically meant that the peak is within ± 0,2 of the vaiue quoted, for example within ± 0.1 of the value quoted.
The invention encompasses polymorphs isolated in pure form or when admixed with other materia!s, for example other polymorphs, or any other materia!.
Thus, in one aspect there is provided a polymorph in isolated or pure form, "isolated" or "pure" form refers to a sample in which the polymorph is present in an amount of >75%, particularly >90%, more particularly >95% and even more particularly >99% relative to other materials which may be present in the sample.
The invention also includes isotopically-labelled compounds, which are identical to the compounds of formula (I) and salts thereof, but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number most commonly found in nature. Examples of isotopes that can be incorporated into the compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen and fluorine, such as 2H, 3H, 11C, 4C and 18F.
The compounds according to formuia (!) may contain one or more asymmetric center (also referred to as a chirai center) and may, therefore, exist as individual enantiomers, diastereomers, or other stereoisomer^ forms, or as mixtures thereof. Chirai centers, such as chirai carbon atoms, may also be present in a substituent such as an aikyl group. Where the stereochemistry of a chira! center present in formula (I), or in any chemical structure illustrated herein, is not specified the structure is intended to encompass any stereoisomer and all mixtures thereof. Thus, compounds according to formula (I) containing one or more chirai center may be used as racemic mixtures, enantiomerically enriched mixtures, or as enantiomerically pure individual stereoisomers. In one embodiment, the invention provides W-II -acryloyl-S-p rrolidinyll-W-tl ^lthiazoloIS^- b]pyridin-2-yi-4-({[1 ,2,2-trimethylpropyl]amino}methy!)-2,6-pyridinediamine in all stereoisomeric forms, and mixtures thereof.
Individual stereoisomers of a compound according to formula (I) which contain one or more asymmetric center may be resolved by methods known to those skilled in the art. For example, such resolution may be carried out (1 ) by formation of diastereoisomeric salts, complexes or other derivatives; (2) by selective reaction with a stereoisomer- specific reagent, for example by enzymatic oxidation or reduction; or (3) by gas-iiquid or liquid chromatography in a chiral enviornment, for example, on a chiral support such as silica with a bound chiral ligand or in the presence of a chiral solvent. The skilled artisan will appreciate that where the desired stereoisomer is converted into another chemical entity by one of the separation procedures described above, a further step is required to liberate the desired form. Alternatively, specific stereoisomers may be synthesized by asymmetric synthesis using optically active reagents, substrates, catalysts or solvents, or by converting one enantiomer to the other by asymmetric transformation.
The compounds according to formula (I) may also contain centers of geometric asymmetry. Where the stereochemistry of a center of geometric asymmetry present in formula (I), or in any chemical structure illustrated herein, is not specified, the structure is intended to encompass the trans geometric isomer, the cis geometric isomer, and all mixtures thereof. Likewise, al! tautomeric forms are a!so included in formula (I) whether such tautomers exist in equilibrium or predominately in one form.
It is to be understood that the references herein to compounds of formula (I) and salts thereof covers the compounds of formula (I) as free acids or free bases, or as salts thereof, for example as pharmaceutically acceptable salts thereof. Thus, in one embodiment, the invention is directed to compounds of formula (!) as the free acid or free base. In another embodiment, the invention is directed to compounds of formula (I) and salts thereof. In a further embodiment, the invention is directed to compounds of formula (I) and pharmaceutically acceptable salts thereof.
The skilled artisan will appreciate that pharmaceuticaliy acceptable salts of the compounds according to formula (I) may be prepared. Indeed, in certain embodiments of the invention, pharmaceutically acceptable salts of the compounds according to formula (I) may be preferred over the respective free base or free acid because such salts impart greater stability or solubility to the molecule thereby facilitating formulation into a dosage form. Accordingly, the invention is further directed to compounds of formula (I) and pharmaceutically acceptable salts thereof. As used herein, the term "pharmaceutically acceptable salts" refers to salts that retain the desired biological activity of the subject compound and exhibit minimal undesired toxicological effects. These pharmaceuticaliy acceptable salts may be prepared in situ during the final isolation and purification of the compound, or by separately reacting the purified compound in its free acid or free base form with a suitable base or acid, respectively. Salts and solvates having non-pharmaceutically acceptable counter-ions or associated solvents are within the scope of the present invention, for example, for use as intermediates in the preparation of other compounds of formula (I) and their pharmaceutically acceptable salts. Thus one embodiment of the invention embraces compounds of formula (I) and salts thereof.
In certain embodiments, compounds according to formula (I) may contain an acidic functional group. Suitable pharmaceuticaiiy-acceptable salts include salts of such acidic functional groups. Representative salts include pharmaceutically acceptable metal salts such as sodium, potassium, lithium, calcium, magnesium, aluminum, and zinc salts; carbonates and bicarbonates of a pharmaceutically acceptable metal cation such as sodium, potassium, lithium, calcium, magnesium, aluminum, and zinc; pharmaceuticaily acceptable organic primary, secondary, and tertiary amines including aliphatic amines, aromatic amines, aliphatic diamines, and hydroxy alkylamines such as methylamine, ethylamine, 2-hydroxyethylamine, diethylamine, TEA, ethyienediamine, ethanolamine, diethanolamine, and cyclohexylamine. in certain embodiments, compounds according to formula (I) may contain a basic functional group and are therefore capable of forming pharmaceutically acceptable acid addition salts by treatment with a suitable acid. Suitable acids include pharmaceutically acceptable inorganic acids and pharmaceutically acceptable organic acids. Representative pharmaceutically acceptable acid addition salts include hydrochloride, hydrobromide, nitrate, methyinitrate, sulfate, bisulfate, sulfamate, phosphate, acetate, hydroxyacetate, phenylacetate, propionate, butyrate, isobutyrate, valerate, maleate, hydroxymaleate, acrylate, fumarate, maiate, tartrate, citrate, salicylate, p- aminosalicyclate, glycollate, lactate, heptanoate, phthalate, oxalate, succinate, benzoate, o-acetoxybenzoate, ch!orobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, naphthoate, hydroxynaphthoate, mandelate, tannate, formate, stearate, ascorbate, palmitate, oleate, pyruvate, pamoate, malonate, laurate, glutarate, glutamate, estolate, methanesulfonate (mesylate), ethanesulfonate (esylate), 2- hydroxyethanesulfonate, benzenesulfonate (besylate), p-aminobenzenesulfonate, p- toluenesulfonate (tosylate), and napthalene-2-sulfonate. In one embodiment, pharmaceutically acceptable acid addition salts include maleate, fumarate, citrate, formate and p-to!uenesulfonate (tosylate).
In one embodiment, the invention provides a compound which is A/-[(3S)-1-acryloyl-3- pyrrolEdiny]]-/V-[1 ,3]thiazolo[5,4-f)]pyridin-2-yl-4-({[(1 S)-1 ,2,2- trimethy!propyl]amino}methyl)-2,6-pyridinediamine or a pharmaceutically acceptable salt thereof.
In a further embodiment, the invention provides a pharmaceutically acceptable salt of N- [(3S)-1 -acryjoyl-3-pyrrolidinyl3-/V-[1 ,3]thiazolo[5,4-/b3pyridin-2-yl-4-({[(1 S)-1 ,2,2^ trimethylpropyl]amino}methyl)-2,6-pyridinediamine,
In one embodiment, the invention provides salts of compounds of formula (!) formed by treatment with fumaric acid, in another embodiment, the invention provides difumarate salts of compounds of formula (I). In a further embodiment, the invention provides Λ/- [{3S)-1-acryloyl-3-pyrrolidinyl]-/v1~[1 ,3]thiazolo[5,4-/b]pyridin-2-y!-4-{{[(1 S)-1 ,2,2- trimethylpropy!]amino}methyl)-2,6-pyrtdinediamine difumarate.
Compound Preparation
The compounds of the invention may be made by a variety of methods, including standard chemistry. Any previously defined variable will continue to have the previously defined meaning unless otherwise indicated. Illustrative general synthetic methods are set out in the following schemes, and can be readily adapted to prepare other compounds of the invention. Specific compounds of the invention are prepared in the Examples section.
Scheme 1
Figure imgf000042_0001
Figure imgf000042_0002
(i) Deprotect if P=protecting gp
(ii) Acyl chloride/base/N P, or
Carboxyiic aci i/HATU/Base/D F
Figure imgf000042_0003
(illustrated for compounds of formula (I) wherein R1 is -CH2OCH3, R2 is -NHR29NHR30, R3 is H, Y is N, wherein R29 is (CH2)n or C3.6cycloalkyi, R30 is -COCHCHR21, -COCH2halo or - COCH2OR22, X is N and P is H or an amine protecting group such as Boc).
It will be appreciated that the genera! approach shown in scheme 1 can be adapted to provide other compounds of formula (!), for instance by using an α,ω-diamine in which either one or both N are incorporated into a 6-membered heterocyclic ring, or by replacing the α,ω-diamine in the penultimate step with for instance (i) an a-hydroxy-co-(N-protected) amino compound (to give a compound of formula (!) wherein R2 is OR9; or (ii) an appropriate a-hydroxy-ω- amino cycloa!kyl compound and then reacting with sulphonyl chloride, to give a compound of formula (I) wherein R8 is C3.6cycloalkyl substituted by - OS02CH3. Scheme 2
Figure imgf000043_0001
Diamine
IPA
(Microwave) or
Diamine
Pailadium catalyst / Ligand
Base
THF
Figure imgf000043_0002
(illustrated for compounds of formula (I) wherein R1 is H, 2-methyi-1 H-imidazoiyl or N- morpholinylCH2, R2 is -NHR29NHR30, R3 is H, Y is CH or N, wherein R29 is (CH2)n or C3. ecyc!oalkyl, R30 is -COCHCHR21 , -COCH2halo or -COCH2OR22, X is N and P is H or an amine protecting group such as Boc).
It will be further appreciated that the general approach shown in scheme 2 can be adapted to provide other compounds of formula (I), by replacing the α,ω-diamine in the penultimate step with for instance (i) an a-hydroxy-to-(N-protected) amino compound (to give a compound of formula i wherein R2 is OR9, or (ii) a phenyl boronic acid optionally substituted by -NR10R11, in the presence of a palladium catalyst, to give a compound of formula (I) wherein R2 is phenyl optionally substituted by -NR10R11, or (iii) an α,ω-diamine in which either one or both N are incorporated into a 6-membered heterocyclic ring. Scheme 3
Figure imgf000044_0001
Be rane
THF
Figure imgf000044_0002
(wherein Ra is MeO or CI, for preparing the starting material for scheme 2 for compounds of formula (I) wherein R1 is N-morpholinylCH2, X is N and Y is N or CH).
It will be appreciated that the general approach shown in scheme 3 can be modified such that the morpholino group is introduced first, followed by amine (using ammonia). Scheme 4
Figure imgf000044_0003
Scheme 5
Figure imgf000044_0004
Borane THF
Figure imgf000044_0005
Figure imgf000045_0001
Figure imgf000045_0002
Compounds of formula (I) wherein R1 to R3, X and Y are as defined above, and salts thereof, may also be prepared by a process comprising final stage modification of one compound of formula (!), or a salt thereof, into another compound of formuia (I), or a salt thereof. Suitable functional group transformations for converting one compound of formula (I) into another compound of formula (!) are weli known in the art and are described in, for instance, Comprehensive Heterocyclic Chemistry II, eds. A. R. Katritzky, C. W. Rees and E. F. V. Scriven (Pergamon Press, 1996), Comprehensive Organic Functional Group Transformations, eds. A.R. Katritzky, O. Meth-Cohn and C.W. Rees (Elsevier Science Ltd., Oxford, 1995), Comprehensive Organic Chemistry, eds. D. Barton and W.D. Ollis (Pergamon Press, Oxford, 1979), and Comprehensive Organic Transformations, R.C. Larock (VCH Publishers Inc., New York, 1989).
Compounds of formula (I) wherein R3 is chloro may be reduced to produce compounds of formula (I) wherein R3 is hydrogen. Suitable reduction conditions include treatment with ammonium formate under microwave irradiation, in the presence of a suitable catalyst such as palladium on activated carbon, in a suitable solvent such as methanol and at a suitable temperature such as about 130 °C.
Compounds of formula (I) wherein R2, R3 and X are as defined above and R1 is - CH2NR6R7 may be prepared from compounds of formula (I) wherein R1 is -CH2OCH3 by conversion of the methoxy group to the corresponding alcohol by treatment with, for example, boron tribromide in the presence of a suitable solvent such as dichloromethane and at a suitable temperature such as ambient temperature, followed by conversion of the alcohol to the corresponding aldehyde by treatment with, for example, manganese dioxide in the presence of a suitable solvent such as tetrahydrofuran and at a suitable temperature such as about 70°C, followed by reaction with an amine of formula R6R7NH2 in the presence of sodium triacetoxyborohydride, a suitable solvent such as dichloromethane and at a suitable temperature such as ambient temperature. Compounds of formula (I) wherein R2, R3 and X are as defined above and R1 is -CH2NH2 may be prepared from the abovementioned aldehyde by conversion to the corresponding oxime by treatment with, for example, hydroxylamine hydrochloride and sodium acetate in the presence of a suitable sovent such as aqueous ethanol and at a suitable temperature such as ambient temperature, followed by treatment with zinc powder in glacial acetic acid at a suitable temperature such as ambient temperature.
Thus, in one embodiment, the invention provides a process for preparing a compound of formula (I) comprising:
a) when R1, R3, X and Y are as defined above and R2 is NR7R8 wherein R7 is as defined above and R8 is -(CH2)nNR12R13, C3.6cycloalkyl substituted by -NR 4R15, or 5- or 6- membered heterocyclyl wherein the 5- or 6-membered heterocyclyl contains a nitrogen atom and is substituted by -R 6, or R7 and R8, together with the nitrogen atom to which they are attached, are linked to form a 6-membered heterocyciyi optionally containing a further nitrogen atom wherein the 6- membered heterocyciyi is substituted by -NR18R19;
reacting a compound of formula (II)
Figure imgf000047_0001
(I f)
wherein R\ R3, X and Y are as defined above and
Y1 is:
-NR7(CH2)nNHR12,
-NR7C3-ecycloalkyl substituted by -NHR14,
-NR7R68 wherein R88 is 5- or 6-membered heterocyciyi which contains a nitrogen atom, a 6-membered heterocyciyi containing, and linked through nitrogen, and optionally containing a further nitrogen atom, wherein the 6-membered heterocyciyi is substituted by -NHR18;
-OR99 wherein R99 is 5- or 6-membered heterocyciyi wherein the 5- or 6-membered heterocyciyi contains a nitrogen atom, or
phenyl wherein the phenyl is substituted by -NHR10 with an acyl chloride compound of formula (l!l):
RaCOCI
(III)
wherein Ra is R11, R13, R15, R16, R19 or R20 are as defined above;
in a suitable solvent such as /V-methy!-2-pyrrolidone, optionaily in the presence of a base such as a tertiary amine such ty/V-diisopropyiethy!amine, and at a suitable temperature, such as ambient temperature, or
with a carboxylic acid of formula (IV):
RaCOOH
(IV)
wherein Ra is R11, R 3, R15, R16, R19or R20 are as defined above,
in the presence of a coupling agent such as HATU, a base and a solvent such as DMF: or
b) when R\ R3, X and Y are as defined above and R2 is -NR7C3-6cyc!oalkyl-S02CH3, reacting a compound of formula (II) as above, and wherein Y1 is -NR7C3.6cycloalkyl substituted by -OH
with methanesulphonyl chloride; or
c) final stage modification of one compound of formula (i), or a salt thereof, into another compound of formula (I), or a salt thereof.
Methods of Use
The compounds of the invention are inhibitors of kinase activity, in particular Itk activity. Compounds which are Itk inhibitors may be useful in the treatment of disorders wherein the underlying pathology is (at least in part) attributable to inappropriate Itk activity, such as asthma. "Inappropriate Itk activity" refers to any Itk activity that deviates from the normal Itk activity expected in a particular patient. Inappropriate Itk may take the form of, for instance, an abnormal increase in activity, or an aberration in the timing and or control of Itk activity. Such inappropriate activity may result then, for example, from overexpression or mutation of the protein kinase leading to inappropriate or uncontrolled activation. Accordingly, in another aspect the invention is directed to methods of treating such disorders.
Such disorders include respiratory diseases including asthma, chronic obstructive pulmonary disease (COPD) and bronchitis; allergic diseases including allergic rhinitis and atopic dermatitis; autoimmune diseases including rheumatoid arthritis, multiple sclerosis, psoriasis, type I diabetes, T cell mediated hypersensitivities, Guillain-Barre Syndrome and Hashimoto's thyroiditis; transplant rejection; graft versus host disease; inflammatory disorders including conjunctivitis, contact dermatitis, inflammatory bowel disease and chronic inflammation; HIV; aplastic anemia; and pain including inflammatory pain.
In one embodiment, the disorder mediated by inappropriate Itk activity is a respiratory disease; an allergic disease; an autoimmune disease; transplant rejection; graft versus host disease; an inflammatory disorder; HIV; aplastic anemia; or pain. In another embodiment, the disorder mediated by inappropriate Itk activity is asthma, chronic obstructive pulmonary disease (COPD), bronchitis, allergic rhinitis, atopic dermatitis, rheumatoid arthritis, multiple sclerosis, psoriasis, type i diabetes, T cell mediated hypersensitivity, Guillain-Barre Syndrome, Hashimoto's thyroiditis, transplant rejection, graft versus host disease, conjunctivitis, contact dermatitis, inflammatory bowel disease, chronic inflammation, HIV, aplastic anemia, or inflammatory pain. In a further embodiment, the disorder mediated by inappropriate Itk activity is asthma. The methods of treatment of the invention comprise administering a safe and effective amount of a compound of formula (I) or a pharmaceuticaliy acceptable sa!t thereof to a patient in need thereof. Individual embodiments of the invention include methods of treating any one of the above-mentioned disorders by administering a safe and effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof to a patient in need thereof.
As used herein, "treat" in reference to a disorder means: (1 ) to ameliorate or prevent the disorder or one or more of the biological manifestations of the disorder, (2) to interfere with (a) one or more points in the biological cascade that leads to or is responsible for the disorder or (b) one or more of the biological manifestations of the disorder, (3) to alleviate one or more of the symptoms or effects associated with the disorder, or (4) to slow the progression of the disorder or one or more of the biological manifestations of the disorder.
As indicated above, "treatment" of a disorder includes prevention of the disorder. The skilled artisan will appreciate that "prevention" is not an absolute term. In medicine, "prevention" is understood to refer to the prophylactic administration of a drug to substantially diminish the likelihood or severity of a disorder or biological manifestation thereof, or to delay the onset of such disorder or biological manifestation thereof.
As used herein, "safe and effective amount" in reference to a compound of formula (I) or a pharmaceutically acceptable salt thereof or other pharmaceutica!ly-active agent means an amount of the compound sufficient to treat the patient's condition but low enough to avoid serious side effects (at a reasonable benefit/risk ratio) within the scope of sound medical judgment. A safe and effective amount of a compound will vary with the particular compound chosen (e.g. consider the potency, efficacy, and half-life of the compound); the route of administration chosen; the disorder being treated; the severity of the disorder being treated; the age, size, weight, and physical condition of the patient being treated; the medical history of the patient to be treated; the duration of the treatment; the nature of concurrent therapy; the desired therapeutic effect; and like factors, but can nevertheless be routinely determined by the skilled artisan.
As used herein, "patient" refers to a human (including adults and children) or other animal, in one embodiment, "patient" refers to a human. The compounds of formula (I) or pharmaceutically acceptable salts thereof may be administered by any suitable route of administration, including both systemic administration and topical administration. Systemic administration includes oral administration, parenteral administration, transdermal administration and recta! administration. Parenteral administration refers to routes of administration other than enteral or transdermal, and is typically by injection or infusion. Parenteral administration includes intravenous, intramuscular, and subcutaneous injection or infusion. Topical administration includes application to the skin as well as intraocular, otic, intravaginal, inhaled and intranasal administration. Inhalation refers to administration into the patient's lungs whether inhaled through the mouth or through the nasal passages. In one embodiment, the compounds of formula (I) or pharmaceutically acceptable salts thereof may be administered orally. In another embodiment, the compounds of formula (I) or pharmaceutically acceptable salts thereof may be administered topically. In another embodiment, the compounds of formula (!) or pharmaceutically acceptable salts thereof may be administered by inhalation. In a further embodiment, the compounds of formula (I) or pharmaceutically acceptable salts thereof may be administered intranasally.
The compounds of formula (I) or pharmaceutically acceptable salts thereof may be administered once or according to a dosing regimen wherein a number of doses are administered at varying intervals of time for a given period of time. For example, doses may be administered one, two, three, or four times per day. In one embodiment, a dose is administered once per day. in a further embodiment, a dose is administered twice per day. Doses may be administered until the desired therapeutic effect is achieved or indefinitely to maintain the desired therapeutic effect. Suitable dosing regimens for a compound of formula (I) or a pharmaceutically acceptable salt thereof depend on the pharmacokinetic properties of that compound, such as absorption, distribution, and half- life, which can be determined by the skilled artisan. In addition, suitable dosing regimens, including the duration such regimens are administered, for a compound of formula (I) or a pharmaceutically acceptable salt thereof depend on the disorder being treated, the severity of the disorder being treated, the age and physical condition of the patient being treated, the medical history of the patient to be treated, the nature of concurrent therapy, the desired therapeutic effect, and like factors within the knowledge and expertise of the skilled artisan. It will be further understood by such skilled artisans that suitable dosing regimens may require adjustment given an individual patient's response to the dosing regimen or over time as individual patient needs change. Typical daily dosages may vary depending upon the particular route of administration chosen. Typical daily dosages for oral administration range from 0.001 mg to 50mg per kg of total body weight, for example from 1 mg to 10mg per kg of total body weight. For example, daily dosages for oral administration may be from Q.5mg to 2g per patient, such as 10mg to 1g per patient.
Additionally, the compounds of formula (I) may be administered as prodrugs. As used herein, a "prodrug" of a compound of formula (I) is a functional derivative of the compound which, upon administration to a patient, eventually liberates the compound of formula (I) in vivo. Administration of a compound of formula (I) as a prodrug may enable the skilled artisan to do one or more of the following: (a) modify the onset of the activity of the compound in vivo; (b) modify the duration of action of the compound in vivo; (c) modify the transportation or distribution of the compound in vivo; (d) modify the solubility of the compound in vivo; and (e) overcome a side effect or other difficulty encountered with the compound. Typical functional derivatives used to prepare prodrugs include modifications of the compound that are chemically or enzymaticaliy cleavable in vivo. Such modifications, which include the preparation of phosphates, amides, esters, thioesters, carbonates, and carbamates, are well known to those skilled in the art. The invention thus provides a method of treating a disorder mediated by inappropriate itk activity comprising administering a safe and effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof to a patient in need thereof. In one embodiment, the invention provides a method of treating a disorder mediated by inappropriate Itk activity comprising administering a safe and effective amount of N-[(ZS)- 1 -acryloyl-S-pyrro!idinylj-W-tl ,33thiazolo[5,4-0]pyridin-2-yi-4-({[(1 S)-1 ,2,2- trimethylpropyi]amino}methyl)-2,6-pyridinediamine or a pharmaceuticaily acceptable salt thereof to a patient in need thereof. In another embodiment, the invention provides a method of treating a disorder mediated by inappropriate Itk activity comprising administering a safe and effective amount of /V-[(3S)-1-acryloyl-3-pyrrolidinyl3-W- [1 ,3]thiazolo[5,4-6]pyridin-2-yl-4-({[{1 S)- ,2,2-trimethylpropyl]amino}methyl)-2,6- pyridinediamine to a patient in need thereof. In a further embodiment, the invention provides a method of treating a disorder mediated by inappropriate Itk activity comprising administering a safe and effective amount of A/-[(3S)-1-acryloyl-3~pyrrolidinyl3-/V- [1 ,3jthiazolo[5,4-0]pyridin-2-yl-4-({[(1 S)-1 ,2,2-trimethy!propyl]amino}methyl)-2,6- pyridinediamine difumarate to a patient in need thereof. In one embodiment, the disorder mediated by inappropriate Itk activity is selected from the group consisting of respiratory diseases (including asthma, chronic obstructive pulmonary disease (COPD) and bronchitis); allergic diseases (including allergic rhinitis and atopic dermatitis); autoimmune diseases (including rheumatoid arthritis, multiple sclerosis, psoriasis, type I diabetes, T cell mediated hypersensitivities, Guillain-Barre Syndrome and Hashimoto's thyroiditis); transplant rejection; graft versus host disease; inflammatory disorders (including conjunctivitis, contact dermatitis, inflammatory bowel disease and chronic inflammation); HIV; aplastic anemia; and pain including inflammatory pain. In one embodiment, the disorder mediated by inappropriate Itk activity is a respiratory disease. In a further embodiment, the disorder mediated by inappropriate Itk activity is asthma. in one embodiment, the invention provides a method of treating a respiratory disease comprising administering a safe and effective amount of /V-[(3S)-1-acryloyl-3-pyrrolidinylj- W-[1 ,3]thiazolo[5,4-0]pyridin-2-yl-4-({[(1 S)-1 ,2,2-trimethylpropyl]amino}methyl)-2,6- pyridinediamine or a pharmaceutically acceptable salt thereof to a patient in need thereof. In another embodiment, the invention provides a method of treating a respiratory disease comprising administering a safe and effective amount of A/-[(3S)-1-acryloy!-3-pyrrolidinyl]- ,3]thiazolo[5,4-/f)jpyridin-2-y!-4-({[(1 S)-1 ,2,2-trimethylpropyl]amino}methyl)-2,6- pyridinediamine to a patient in need thereof. In a further embodiment, the invention provides a method of treating a respiratory disease comprising administering a safe and effective amount of W-[(3S)-1 -acryloy!-3-pyrrolidinyl]-W-[1 ,3]thiazolo[5,4-b]pyridin-2-yl-4- ({[(1 S)-1 ,2,2-trimethylpropyl]amino}methyl)-2,6-pyridtnediamine difumarate to a patient in need thereof.
In one embodiment, the invention provides a method of treating asthma comprising administering a safe and effective amount of W-[(3S)-1-acry!oyl-3-pyrroiidinyl]-/v\ [1 ,3]thiazolo[5,4-/bjpyridin-2-yl-4-({[(1 S)-1 ,2,2-trimethyipropyl]amino}methyl)-2,6- pyridinediamine or a pharmaceutically acceptable salt thereof to a patient in need thereof. In another embodiment, the invention provides a method of treating asthma comprising administering a safe and effective amount of W-[{3S)-1-acryloyl-3-pyrrolidinyl]-/V- [1 ,3]thiazo!o[5,4-0]pyridin-2-yl-4-({[(1 S)-1 ,2,2-trimethylpropy!]amino}methyi)-2,6- pyridinediamine to a patient in need thereof. In a further embodiment, the invention provides a method of treating asthma comprising administering a safe and effective amount of W-[(3S)-1 -acryloyl-3-pyrrolidinyl]-A "-[1 ,3]thiazolo[5,4- >]pyridin-2-yl~4-({[(1 S)- l ^^-tnmethylpropyljaminoJmethy ^^-pyridinediarnine difumarate to a patient in need thereof.
!n one embodiment, the invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in medical therapy. In another embodiment, the invention provides V-f(3S)-1 -acryloyl-3-pyrrolidinyl)-/V-[1 ,3]thia2oSot5,4- 6]pyridin-2-y!-4-({[(1 S)-1 ,2,2-trimethylpropyl]amino}methyl)-2,6-pyridinediamine or a pharmaceutically acceptable salt thereof for use in medical therapy, !n another embodiment, the invention provides A/-[(3S)-1-acryloyl-3-pyrrolidinyl]-A/,-[1 ,3]thiazolo[5,4- 0]pyridin-2-yl-4-{{[(1 S)-1 ,2,2-trimethylpropyi]amino}methyl)-2,6-pyhdinediamine for use in medical therapy, in a further embodiment, the invention provides A/-[(3S)-1-acryioyi-3- pyrrolidinyll-Af-tl ^lthiazolotS^-dlpyridin^-y -intlSJ-l^^- thmethylpropyl]amino}methyl)-2,6-pyridinediamine difumarate for use in medical therapy. In one embodiment, the invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in the treatment of a disorder mediated by inappropriate Itk activity. In another embodiment, the invention provides W-[(3S)-1- acryloy!-3-pyrroiidinyl]-W-[1 ,3]th!azolo[5,4-0]pyridin-2-y!-4-({[{1 S)-1 ,2,2- trimethylpropyl]amino}methyl)-2,6-pyridinediamine or a pharmaceutically acceptable salt thereof for use in the treatment of a disorder mediated by inappropriate Itk activity. In another embodiment, the invention provides W-[(3S)-1-acryloyi-3-pyrrolidiny!]-A/'- [1 ,3]thiazo!o[5,4-jb]pyridin-2-yl-4-({[{1 S)-1 ,2,2-trimethylpropyS]amino}methy!)-2,6- pyridinediamine for use in the treatment of a disorder mediated by inappropriate Itk activity. In a further embodiment, the invention provides A/-[(3S)-1-acryloyl-3-pyrrolidinyl]- ,33thiazolo[5,4- )]pyridin-2-yl-4-({[{1 S)-1 ,2,2-trimethy!propyl]amino}methyl)-2,6- pyridinediamine difumarate for use in the treatment of a disorder mediated by inappropriate Itk activity.
In one embodiment, the invention provides the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for use in the treatment of a disorder mediated by inappropriate Itk activity. In another embodiment, the invention provides the use of A/-[(3S)-1-acryloyl-3-pyrrolidinyl3-/V-[1 ,3]thiazolo[5>4- jt>jpyridin-2-yl-4-({[(1 S)-1 ,2,2-tnmethylpropyl]amino}methyl)-2,6-pyhdinediamine or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for use in the treatment of a disorder mediated by inappropriate itk activity. In another embodiment, the invention provides the use of A/-[(3S)-1 -acryloyl-3-pyrrolidinyl]-A '-[1 ,3]thiazolo[5,4- jb]pyridin-2-yl-4-({[(1 S)-1 ^^-tnmethylpropyljaminojmethyl^.e-pyridinediamine in the manufacture of a medicament for use in the treatment of a disorder mediated by inappropriate Itk activity, in a further embodiment, the invention provides the use of N- [(3S)-1-acry!oyl-3-pyrrolidinyl]-A/,-[1 ,3]thiazolo[5,4-j ]pyridin-2-yl-4-({[(1 S)-1 ,2,2- trimethyipropy!]amino}methyl)-2,6-pyridinediamine difumarate in the manufacture of a medicament for use in the treatment of a disorder mediated by inappropriate Itk activity.
In one embodiment, the invention provides a pharmaceutical composition for the treatment or prophylaxis of a disorder mediated by inappropriate Itk activity comprising a compound of formula (I) or a pharmaceuticaliy acceptable salt thereof. In another embodiment, the invention provides a pharmaceutical composition for the treatment or prophylaxis of a disorder mediated by inappropriate Itk activity comprising W-[(3S)-1- acryloyl-3-pyrroiidinyl]-/V-[1 ,3]thiazolo[5,4-0]pyridin-2-y!-4-({[(1 S)-1 ,2,2- trimethylpropyl]amino}methyi)-2,6-pyridinediamine or a pharmaceutically acceptable salt thereof. In another embodiment, the invention provides a pharmaceutical composition for the treatment or prophylaxis of a disorder mediated by inappropriate Itk activity comprising W-[(3S)-1-acryloyl-3-pyrrolidinyi]-W-[1 ,3]thtazolo[5,4-i)]pyridin-2-yi-4-{{[(1 S)- ,2,2- trimethy!propyl]amino}methyl)-2,6-pyridinediamine. In a further embodiment, the invention provides a pharmaceutical composition for the treatment or prophylaxis of a disorder mediated by inappropriate Itk activity comprising W-[(3S)-1 -acryloyl-3-pyrrolidinyl]- v1- [1 ,3]thiazolo[5,4-0]pyridin-2-yl-4-({[(1 S)-1 ,2,2-trimethylpropyl]amino}methyl)-2,6- pyridinediamine difumarate.
Compositions
The compounds of formula (I) and pharmaceutically acceptable salts thereof will normally, but not necessarily, be formulated into pharmaceutical compositions prior to administration to a patient.
Accordingly, in another aspect the invention is directed to pharmaceutical compositions comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable excipients.
In one embodiment, the invention provides a pharmaceutical composition comprising N- [(SSH-acryioy!-S-pyrrolidiny!l-W-EI ,3]thiazolo[5,4-6]pyridin-2-yl-4-({[(1 S)-1 ,2,2- thmethylpropyi3amino}methyl)-2,6-pyridinediamine or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable excipients. In another embodiment, the invention provides a pharmaceutical composition comprising W-[(3S)-1-acryloyl-3-pyrrolidinyl]-W-[1 ,33thiazolo[5,4-/3]pyridin-2-yl-4-({[(1 S)-1 ,2,2- trimethylpropyl]amino}methyi)-2,6-pyridinediamine and one or more pharmaceutically acceptable excipients.
In a further embodiment, the invention provides a pharmaceutical composition comprising W-[{3S)-1-acryloyl-3-pyrrolidinyl]-/V-[1 ,3]thiazolo[5,4-6]pyridin-2-yl-4-({[(1 S)-1 ,2,2- trimethylpropyl]amino}methyl)-2,6-pyridinediamine difumarate and one or more pharmaceutically acceptable excipients.
In a further aspect the invention is directed to pharmaceutical compositions for the treatment or prophylaxis of a disorder mediated by inappropriate Itk activity comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof. The pharmaceutical compositions of the invention may be prepared and packaged in bulk form wherein a safe and effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof can be extracted and then given to the patient such as with powders or syrups. Alternatively, the pharmaceutical compositions of the invention may be prepared and packaged in unit dosage form wherein each physically discrete unit contains a compound of formula (I) or a pharmaceutically acceptable salt thereof. When prepared in unit dosage form, the pharmaceutical compositions of the invention typically may contain, for example, from 0.5mg to 1 g, or from 1 mg to 700mg, or from 5mg to 100mg of a compound of formula (I) or a pharmaceutically acceptable salt thereof.
The pharmaceutical compositions of the invention typically contain one compound of formula (I) or a pharmaceutically acceptable salt thereof.
As used herein, "pharmaceutically acceptable excipient" means a pharmaceutically acceptable material, composition or vehicle involved in giving form or consistency to the pharmaceutical composition. Each excipient must be compatible with the other ingredients of the pharmaceutical composition when commingled such that interactions which would substantially reduce the efficacy of the compound of formula (I) or a pharmaceutically acceptable salt thereof when administered to a patient and interactions which would result in pharmaceutical compositions that are not pharmaceutically acceptable are avoided. In addition, each excipient must of course be pharmaceutically acceptable eg of sufficiently high purity.
The compound of formula (I) or a pharmaceutically acceptable salt thereof and the pharmaceutically acceptable excipient or excipients will typically be formulated into a dosage form adapted for administration to the patient by the desired route of administration. For example, dosage forms include those adapted for (1 ) oral administration such as tablets, capsules, caplets, piils, troches, powders, syrups, elixers, suspensions, solutions, emulsions, sachets, and cachets; (2) parenteral administration such as sterile solutions, suspensions, and powders for reconstitution; (3) transdermal administration such as transdermal patches; (4) rectal administration such as suppositories; (5) inhalation such as aerosols, solutions, and dry powders; and (6) topical administration such as creams, ointments, lotions, solutions, pastes, sprays, foams, and gels.
Suitable pharmaceutically acceptable excipients will vary depending upon the particular dosage form chosen. In addition, suitable pharmaceutically acceptable excipients may be chosen for a particular function that they may serve in the composition. For example, certain pharmaceutically acceptable excipients may be chosen for their ability to facilitate the production of uniform dosage forms. Certain pharmaceutically acceptable excipients may be chosen for their ability to facilitate the production of stable dosage forms. Certain pharmaceutically acceptable excipients may be chosen for their ability to facilitate the carrying or transporting of the compound or compounds of formula (I) or pharmaceutically acceptable salts thereof once administered to the patient from one organ, or portion of the body, to another organ, or portion of the body. Certain pharmaceutically acceptable excipients may be chosen for their ability to enhance patient compliance.
Suitable pharmaceuticaliy-acceptabie excipients include the following types of excipients: Diluents, fillers, binders, disintegrants, lubricants, glidants, granulating agents, coating agents, wetting agents, solvents, co-solvents, suspending agents, emulsifiers, sweetners, flavoring agents, flavor masking agents, coloring agents, anticaking agents, hemectants, chelating agents, plasticizers, viscosity increasing agents, antioxidants, preservatives, stabilizers, surfactants, and buffering agents. The skilled artisan will appreciate that certain pharmaceuticaliy-acceptabie excipients may serve more than one function and may serve alternative functions depending on how much of the excipient is present in the formulation and what other excipients are present in the formulation. Skilled artisans possess the knowledge and skill in the art to enable them to select suitable pharmaceutically-acceptable excipients in appropriate amounts for use in the invention. In addition, there are a number of resources that are available to the skilled artisan which describe pharmaceutically-acceptable excipients and may be useful in selecting suitable pharmaceutically-acceptable excipients. Examples include Remington's Pharmaceutical Sciences (Mack Publishing Company), The Handbook of Pharmaceutical Additives (Gower Publishing Limited), and The Handbook of Pharmaceutical Excipients (the American Pharmaceutical Association and the Pharmaceutical Press).
The pharmaceutical compositions of the invention are prepared using techniques and methods known to those skilled in the art. Some of the methods commonly used in the art are described in Remington's Pharmaceutical Sciences (Mack Publishing Company). Accordingly, in another aspect the invention is directed to process for the preparation of a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof and one or more pharmaceutically-acceptable excipients which comprises mixing the ingredients. A pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof may be prepared by, for example, admixture at ambient temperature and atmospheric pressure.
In one embodiment, the compounds of formula (!) or pharmaceutically acceptable salts thereof will be formulated for oral administration. In another embodiment, the compounds of formula (I) or pharmaceutically acceptable salts thereof will be formulated for inhaled administration. In a further embodiment, the compounds of formula (I) or pharmaceutically acceptable salts thereof will be formulated for intranasal administration.
In one aspect, the invention is directed to a solid oral dosage form such as a tablet or capsule comprising a safe and effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof and a diluent or filler. Suitable diluents and fillers include lactose, sucrose, dextrose, mannitol, sorbitol, starch (e.g. corn starch, potato starch, and pre-gelatinized starch), cellulose and its derivatives (e.g. microcrystalline cellulose), calcium sulfate, and dibasic calcium phosphate. The oral solid dosage form may further comprise a binder. Suitable binders include starch (e.g. corn starch, potato starch, and pre-geiatinized starch), gelatin, acacia, sodium alginate, alginic acid, tragacanth, guar gum, povidone, and celiuiose and its derivatives (e.g. microcrystalline cellulose). The oral solid dosage form may further comprise a disintegrant. Suitable disintegrants include crospovidone, sodium starch glycolate, croscarmelose, alginic acid, and sodium carboxymethyl cellulose. The oral solid dosage form may further comprise a lubricant. Suitable lubricants include stearic acid, magnesuim stearate, calcium stearate, and talc.
Where appropriate, dosage unit formulations for oral administration can be microencapsulated. The composition can also be prepared to prolong or sustain the release as for example by coating or embedding particulate material in polymers, wax or the like.
The compounds of formula (I) or pharmaceutically acceptable salts thereof may also be coupled with soluble polymers as targetabie drug carriers. Such polymers can include polyvinylpyrrolidone, pyran copolymer, polyhydroxypropylmethacrylamide -phenol, po!yhydroxyethy!aspartamidephenol, or poiyethyleneoxidepolylysine substituted with palmitoyl residues. Furthermore, the compounds of formula (I) or pharmaceutically acceptable salts thereof may be coupled to a class of biodegradable polymers useful in achieving controlled release of a drug, for example, polylactic acid, polepsilon caproiactone, polyhydroxy butyric acid, polyorthoesters, polyacetals, polydihydropyrans, poiycyanoacry!ates and cross-linked or amphipathic block copolymers of hydrogels.
In another aspect, the invention is directed to a liquid oral dosage form. Oral liquids such as solution, syrups and elixirs can be prepared in dosage unit form so that a given quantity contains a predetermined amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof. Syrups can be prepared by dissolving the compound of formula (I) or a pharmaceutically acceptable salt thereof in a suitably flavored aqueous solution, while elixirs are prepared through the use of a non-toxic alcoholic vehicle. Suspensions can be formulated by dispersing the compound of formula (I) or a pharmaceutically acceptable salt thereof in a non-toxic vehicle. So!ubilizers and emulsifiers such as ethoxyiated isostearyl alcohols and polyoxy ethylene sorbitol ethers, preservatives, flavor additive such as peppermint oil or natural sweeteners or saccharin or other artificial sweeteners, and the like can also be added.
In another aspect, the invention is directed to a dosage form adapted for administration to a patient by inhalation. For example, as a dry powder, an aerosol, a suspension, or a solution composition. Dry powder compositions for delivery to the lung by inhalation typically comprise a compound of formula (i) or a pharmaceutically acceptable salt thereof as a finely divided powder together with one or more pharmaceuticaily-acceptable excipients as finely divided powders. Pharmaceuticaily-acceptable excipients particularly suited for use in dry powders are known to those skilled in the art and include lactose, starch, mannitol, and mono-, di-, and polysaccharides. The finely divided powder may be prepared by, for example, micronisation and milling. Generally, the size-reduced (eg micronised) compound can be defined by a D50 value of about 1 to about 10 microns (for example as measured using laser diffraction).
The dry powder may be administered to the patient via a reservoir dry powder inhaler (RDPI) having a reservoir suitable for storing multiple (un-metered doses) of medicament in dry powder form. RDPIs typically include a means for metering each medicament dose from the reservoir to a delivery position. For example, the metering means may comprise a metering cup, which is movable from a first position where the cup may be filled with medicament from the reservoir to a second position where the metered medicament dose is made available to the patient for inhalation. Alternatively, the dry powder may be presented in capsules (e.g. gelatin or plastic), cartridges, or blister packs for use in a multi-dose dry powder inhaler (MDPl). MDPIs are inhalers wherein the medicament is comprised within a multi-dose pack containing (or otherwise carrying) multiple defined doses (or parts thereof) of medicament. When the dry powder is presented as a blister pack, it comprises multiple blisters for containment of the medicament in dry powder form. The blisters are typically arranged in regular fashion for ease of release of the medicament therefrom. For example, the blisters may be arranged in a generally circular fashion on a disc-form blister pack, or the blisters may be elongate in form, for example comprising a strip or a tape. Each capsule, cartridge, or blister may, for example, contain between 20μg-10mg of the compound of formula (I) or a pharmaceutically acceptable salt thereof.
Aerosols may be formed by suspending or dissolving a compound of formula (I) or a pharmaceutically acceptable salt thereof in a liquified propellant. Suitable propellants include halocarbons, hydrocarbons, and other liquified gases. Representative propellants include: trichlorofluoromethane (propellant 1 1), dichlorofluoromethane (propellant 12), dichlorotetrafluoroethane (propellant 114), tetraf!uoroethane (HFA-134a), 1 ,1 - dif!uoroethane (HFA-152a), difluoromethane (HFA-32), pentafluoroethane (HFA-12), heptaf!uoropropane (HFA-227a), perfluoropropane, perfluorobutane, perfluoropentane, butane, isobutane, and pentane. Aerosols comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof will typically be administered to a patient via a metered dose inhaler (MD!). Such devices are known to those skilled in the art.
The aerosol may contain additional pharmaceutical!y-acceptabie excipients typically used with MDIs such as surfactants, lubricants, cosoivents and other excipients to improve the physical stability of the formulation, to improve valve performance, to improve solubility, or to improve taste.
There is thus provided as a further aspect of the invention a pharmaceutical aerosol formulation comprising a compound of formula (i) or a pharmaceutically acceptable salt thereof and a fluorocarbon or hydrogen-containing ch!orofluorocarbon as propellant, optionally in combination with a surfactant and/or a cosolvent,
According to another aspect of the invention, there is provided a pharmaceutical aerosol formulation wherein the propellant is selected from 1 ,1 ,1 ,2-tetrafluoroethane, 1 ,1 , 1 ,2,3,3,3-heptafluoro-n-propane and mixtures thereof.
The formulations of the invention may be buffered by the addition of suitable buffering agents.
Capsules and cartridges for use in an inhaler or insufflator, of for example gelatine, may be formulated containing a powder mix for inhalation of a compound of formula (I) or a pharmaceutically acceptable salt thereof and a suitable powder base such as lactose or starch. Each capsule or cartridge may generally contain from 20μ9 to 10mg of the compound of formula (I) or pharmaceutically acceptable salt thereof. Alternatively, the compound of formula (I) or pharmaceutically acceptable salt thereof may be presented without excipients such as lactose.
The proportion of the active compound of formula (I) or pharmaceutically acceptable salt thereof in the local compositions according to the invention depends on the precise type of formulation to be prepared but will generally be within the range of from 0.001 to 10% by weight. Generally, for most types of preparations, the proportion used will be within the range of from 0.005 to 1 %, for example from 0.01 to 0.5%. However, in powders for inhalation or insufflation the proportion used will normally be within the range of from 0.1 to 5%.
Aerosol formulations are preferably arranged so that each metered dose or "puff of aerosol contains from 20 g to 10mg, preferably from 20μg to 2000μg> more preferably from about 20μ9 to 500μg of a compound of formula (I). Administration may be once daily or several times daily, for example 2, 3, 4 or 8 times, giving for example 1 , 2 or 3 doses each time. The overall daily dose with an aerosol will be within the range from 100μg to 10mg, preferably from 200μg to 2000μg. The overall daily dose and the metered dose delivered by capsules and cartridges in an inhaler or insufflator will generally be double that delivered with aerosol formulations.
In the case of suspension aerosol formulations, the particle size of the particulate (e.g., mtcronised) drug should be such as to permit inhalation of substantially all the drug into the lungs upon administration of the aerosol formulation and will thus be less than 100 microns, desirably less than 20 microns, and in particular in the range of from 1 to 10 microns, such as from 1 to 5 microns, more preferably from 2 to 3 microns.
The formulations of the invention may be prepared by dispersal or dissolution of the medicament and a compound of formula (I) or a pharmaceutically acceptable salt thereof in the selected prope!lant in an appropriate container, for example, with the aid of sonication or a high-shear mixer. The process is desirably carried out under controlled humidity conditions. The chemical and physical stability and the pharmaceutical acceptability of the aerosol formulations according to the invention may be determined by techniques well known to those skilled in the art. Thus, for example, the chemical stability of the components may be determined by HPLC assay, for example, after prolonged storage of the product. Physical stability data may be gained from other conventional analytical techniques such as, for example, by leak testing, by valve delivery assay (average shot weights per actuation), by dose reproducibility assay (active ingredient per actuation) and spray distribution analysis.
The stability of the suspension aerosol formulations according to the invention may be measured by conventional techniques, for example, by measuring flocculation size distribution using a back light scattering instrument or by measuring particle size distribution by cascade impaction or by the "twin impinger" analytical process. As used herein reference to the "twin impinger" assay means "Determination of the deposition of the emitted dose in pressurised inhalations using apparatus A" as defined in British Pharmacopaeia 1988, pages A204-207, Appendix XVII C. Such techniques enable the "respirable fraction" of the aerosol formulations to be calculated. One method used to calculate the "respirable fraction" is by reference to "fine particle fraction" which is the amount of active ingredient collected in the lower impingement chamber per actuation expressed as a percentage of the total amount of active ingredient delivered per actuation using the twin impinger method described above.
The term "metered dose inhaler" or MDI means a unit comprising a can, a secured cap covering the can and a formulation metering valve situated in the cap. MDI system includes a suitable channelling device. Suitable channelling devices comprise for example, a valve actuator and a cylindrical or cone-like passage through which medicament may be delivered from the filled canister via the metering valve to the nose or mouth of a patient such as a mouthpiece actuator.
MDI canisters generally comprise a container capable of withstanding the vapour pressure of the propellant used such as a plastic or plastic-coated glass bottle or preferably a metal can, for example, aluminium or an alloy thereof which may optionally be anodised, lacquer-coated and/or plastic-coated (for example incorporated herein by reference WO96/32099 wherein part or all of the internal surfaces are coated with one or more fluorocarbon polymers optionally in combination with one or more non-fluorocarbon polymers), which container is closed with a metering valve. The cap may be secured onto the can via ultrasonic welding, screw fitting or crimping. MDIs taught herein may be prepared by methods of the art (e.g. see Byron, above and WO96/32099). Preferably the canister is fitted with a cap assembly, wherein a drug-metering valve is situated in the cap, and said cap is crimped in place. In one embodiment of the invention the metallic internal surface of the can is coated with a fluoropolymer, more preferably blended with a non-fluoropolymer. In another embodiment of the invention the metailic internal surface of the can is coated with a polymer blend of polytetrafluoroethyiene (PTFE) and polyethersulfone (PES). In a further embodiment of the invention the whole of the metallic internal surface of the can is coated with a polymer blend of polytetrafluoroethyiene (PTFE) and polyethersulfone (PES). The metering valves are designed to deliver a metered amount of the formu!ation per actuation and incorporate a gasket to prevent leakage of propellant through the valve. The gasket may comprise any suitable elastomeric materia! such as, for example, Sow density polyethylene, chlorobutyl, bromobutyl, EPDM, black and white butadiene- acrylonitrile rubbers, butyl rubber and neoprene, Suitable valves are commercially available from manufacturers well known in the aerosol industry, for example, from Valois, France (e.g. DF10, DF30, DF60), Bespak pic, UK (e.g. BK300, BK357) and 3M-
TM
Neotechnic Ltd, UK (e.g. Spraymiser ). In various embodiments, the DIs may also be used in conjunction with other structures such as, without limitation, overwrap packages for storing and containing the MD!s, including those described in U.S. Patent Nos. 6,1 19,853; 6, 179,1 18; 6,315,112; 6,352,152; 6,390,291 ; and 6,679,374, as well as dose counter units such as, but not limited to, those described in U.S. Patent Nos. 6,360,739 and 6,431 ,168.
Conventional bulk manufacturing methods and machinery well known to those skilled in the art of pharmaceutical aerosol manufacture may be employed for the preparation of large-scale batches for the commercial production of filled canisters. Thus, for example, in one bulk manufacturing method for preparing suspension aerosol formulations a metering valve is crimped onto an aluminium can to form an empty canister. The particulate medicament is added to a charge vessel and liquefied propellant together with the optional excipients is pressure filled through the charge vessel into a manufacturing vessel. The drug suspension is mixed before recirculation to a filling machine and an aliquot of the drug suspension is then filled through the metering valve into the canister. In one example bulk manufacturing method for preparing solution aerosol formulations a metering valve is crimped onto an aluminium can to form an empty canister. The liquefied propellant together with the optional excipients and the dissolved medicament is pressure filled through the charge vessel into a manufacturing vessel. In an alternative process, an aliquot of the liquefied formulation is added to an open canister under conditions which are sufficiently cold to ensure the formulation does not vaporise, and then a metering valve crimped onto the canister.
Typically, in batches prepared for pharmaceutical use, each filled canister is check- weighed, coded with a batch number and packed into a tray for storage before release testing. Suspensions and solutions comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof may also be administered to a patient via a nebulizer. The solvent or suspension agent utilized for nebulization may be any pharmaceutically-acceptable liquid such as water, aqueous saline, alcohols or glycols, e.g., ethanol, isopropylalcohol, glycerol, propylene glycol, polyethylene glycol, etc. or mixtures thereof. Saline solutions utilize salts which display little or no pharmacological activity after administration. Both organic salts, such as alkali metal or ammonium halogen salts, e.g., sodium chloride, potassium chloride or organic salts, such as potassium, sodium and ammonium salts or organic acids, e.g., ascorbic acid, citric acid, acetic acid, tartaric acid, etc. may be used for this purpose.
Other pharmaceutically-acceptable excipients may be added to the suspension or solution, The compound of formula (I) or pharmaceutically acceptable salt thereof may be stabilized by the addition of an inorganic acid, e.g., hydrochloric acid, nitric acid, sulphuric acid and/or phosphoric acid; an organic acid, e.g., ascorbic acid, citric acid, acetic acid, and tartaric acid, etc., a complexing agent such as EDTA or citric acid and salts thereof; or an antioxidant such as antioxidant such as vitamin E or ascorbic acid. These may be used alone or together to stabilize the compound of formula (I) or pharmaceutically acceptable salt thereof. Preservatives may be added such as benzalkonium chloride or benzoic acid and salts thereof. Surfactant may be added particularly to improve the physical stability of suspensions. These include lecithin, disodium dioctylsulphosuccinate, oleic acid and sorbitan esters.
In a further aspect, the invention is directed to a dosage form adapted for intranasal administration.
Formulations for administration to the nose may include pressurised aerosol formulations and aqueous formulations administered to the nose by pressurised pump. Formulations which are non-pressurised and adapted to be administered topically to the nasal cavity are of particular interest. Suitable formulations contain water as the diluent or carrier for this purpose. Aqueous formulations for administration to the lung or nose may be provided with conventional excipients such as buffering agents, tonicity modifying agents and the like. Aqueous formulations may also be administered to the nose by nebulisation. The compounds of formula (I) or pharmaceuticaily acceptable salts thereof may be formulated as a fluid formulation for delivery from a fluid dispenser, for example a fluid dispenser having a dispensing nozzie or dispensing orifice through which a metered dose of the fluid formulation is dispensed upon the application of a user-applied force to a pump mechanism of the fluid dispenser. Such fluid dispensers are generally provided with a reservoir of multiple metered doses of the fluid formulation, the doses being dispensable upon sequential pump actuations. The dispensing nozzle or orifice may be configured for insertion into the nostrils of the user for spray dispensing of the fluid formulation into the nasal cavity. A fluid dispenser of the aforementioned type is described and illustrated in WO05/044354, the entire content of which is hereby incorporated herein by reference. The dispenser has a housing which houses a fluid discharge device having a compression pump mounted on a container for containing a fluid formulation. The housing has at least one finger-operable side lever which is movable inwardly with respect to the housing to cam the container upwardly in the housing to cause the pump to compress and pump a metered dose of the formulation out of a pump stem through a nasal nozzle of the housing. In one embodiment, the fluid dispenser is of the general type illustrated in Figures 30-40 of WO05/044354.
Pharmaceutical compositions adapted for intranasal administration wherein the carrier is a solid include a coarse powder having a particle size for example in the range 20 to 500 microns which is administered by rapid inhalation through the nasai passage from a container of the powder held close up to the nose. Suitable compositions wherein the carrier is a liquid, for administration as a nasal spray or as nasal drops, include aqueous or oil solutions of the compound of formula (I) or a pharmaceutically acceptable salt thereof.
Pharmaceutical compositions adapted for transdermal administration may be presented as discrete patches intended to remain in intimate contact with the epidermis of the patient for a prolonged period of time. For example, the active ingredient may be delivered from the patch by iontophoresis as generally described in Pharmaceutical Research, 3(6), 318 (1986).
Pharmaceutical compositions adapted for topical administration may be formulated as ointments, creams, suspensions, lotions, powders, solutions, pastes, gels, sprays, aerosols or oils. Ointments, creams and gels, may, for example, be formulated with an aqueous or oily base with the addition of suitable thickening and/or gelling agent and/or solvents. Such bases may thus, for example, include water and/or an oil such as liquid paraffin or a vegetable oil such as arachis oil or castor oil, or a solvent such as polyethylene glycol. Thickening agents and gelling agents which may be used according to the nature of the base include soft paraffin, aluminium stearate, cetosteary! aicohol, polyethylene glycols, woolfat, beeswax, carboxypo!ymethyiene and cellulose derivatives, and/or glyceryl monostearate and/or non-ionic emulsifying agents. Lotions may be formulated with an aqueous or oily base and will in general also contain one or more emulsifying agents, stabilising agents, dispersing agents, suspending agents or thickening agents.
Powders for externa! application may be formed with the aid of any suitable powder base, for example, talc, lactose or starch. Drops may be formulated with an aqueous or nonaqueous base also comprising one or more dispersing agents, solubilising agents, suspending agents or preservatives.
Topical preparations may be administered by one or more applications per day to the affected area; over skin areas occlusive dressings may advantageously be used. Continuous or prolonged delivery may be achieved by an adhesive reservoir system.
For treatments of the eye or other external tissues, for example mouth and skin, the compositions may be applied as a topical ointment or cream. When formulated in an ointment, the compound of formula (I) or a pharmaceutically acceptable salt thereof may be employed with either a paraffinic or a water-miscible ointment base. Alternatively, the compound of formula (i) or pharmaceutically acceptable salt thereof may be formulated in a cream with an oil-in-water cream base or a water-in-oil base. Pharmaceutical compositions adapted for parenteral administration include aqueous and non-aqueous sterile injection solutions which may contain anti-oxidants, buffers, bacteriostats and solutes which render the formulation isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions which may include suspending agents and thickening agents. The compositions may be presented in unit- dose or multi-dose containers, for example sealed ampoules and vials, and may be stored in a freeze-dried (lyophilized) condition requiring only the addition of the sterile liquid carrier, for example water for injections, immediately prior to use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules and tablets.
The compound and pharmaceutical formulations according to the invention may be used in combination with or include one or more other therapeutic agents, for example selected from anti-inflammatory agents, anticholinergic agents {particularly an bA M2/Mi receptor antagonist), 32-adrenoreceptor agonists, antiinfective agents, such as antibiotics or antivirals, or antihistamines. The invention thus provides, in a further aspect, a combination comprising a compound of formula (!) or a pharmaceutically acceptable salt thereof together with one or more other therapeutically active agents, for example selected from an anti-inflammatory agent, such as a corticosteroid or an NSAID, an anticholinergic agent, a 2-adrenoreceptor agonist, an antiinfective agent, such as an antibiotic or an antiviral, or an antihistamine. One embodiment of the invention encompasses combinations comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof together with a 2-3 ΓθηοΓθθθ ίθΓ agonist, and/or an anticholinergic, and/or a PDE-4 inhibitor, and/or an antihistamine.
One embodiment of the invention encompasses combinations comprising one or two other therapeutic agents.
It will be clear to a person skilled in the art that, where appropriate, the other therapeutic ingredient(s) may be used in the form of salts, for example as alkali metal or amine salts or as acid addition salts, or prodrugs, or as esters, for example lower alky! esters, or as solvates, for example hydrates to optimise the activity and/or stability and/or physical characteristics, such as solubility, of the therapeutic ingredient. It will be clear also that, where appropriate, the therapeutic ingredients may be used in optically pure form.
In one embodiment, the invention encompasses a combination comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof together with a p2-adrenoreceptor agonist.
Examples of 2-adrenoreceptor agonists include salmeterol (which may be a racemate or a single enantiomer such as the R-enantiomer), salbutamol (which may be a racemate or a single enantiomer such as the R-enantiomer), formoterol (which may be a racemate or a single duastereomer such as the R. R-diastereomer), salmefamol, fenoterol carmotero!, etanterol, naminterol, clenbuterol, pirbuterol, flerbuterol, reproteroi, bambuterol, indacaterol, terbutaline and salts thereof, for example the xinafoate (1 -hydroxy-2- naphthalenecarboxy!ate) salt of salmeterol, the sulfate salt or free base of salbutamol or the fumarate salt of formoterol. !n one embodiment, long-acting p2-adrenoreceptor agonists, for example, compounds which provide effective bronchodilation for about 12 hrs or longer, are preferred.
Other P2-adrenoreceptor agonists include those described in WO 02/066422, WO
02/070490, WO 02/076933, WO 03/024439, WO 03/072539, WO 03/091204, WO 04/016578, WO 2004/022547, WO 2004/037807, WO 2004/037773, WO 2004/037768, WO 2004/039762, WO 2004/039766, WO01/42193 and WO03/042160.
Examples of β2^ re no receptor agonists include:
3- (4-{[6-({(2R)-2-hydroxy-2-[4-hydroxy-3-(hydroxymethyl)phenyl]ethy!}amino)
hexyl] oxy} butyl) benzenesulfonamide;
3-(3-{[7-({{2R)-2-hydroxy-2-[4-hydroxy-3-hydroxymethyl) phenyl] ethyl}-amino) heptyl] oxy} propyl) benzenesulfonamide;
4- {(1 R)-2-[(6-{2-[(2, 6-dichlorobenzyl) oxy] ethoxy} hexyl) amino]-1-hydroxyethyl}-2- (hydroxymethyl) phenol;
4-{(1 R)-2-[(6-{4-[3-(cyclopentylsulfonyl)phenyl]butoxy}hexyl)amino]-1-hydroxyethyl}-2- (hydroxymethyl)phenol;
N-[2-hydroxyl-5-[(1 R)-1 -hydroxy-2-[[2-4-[[(2R)-2-hydroxy-2- phenylethyl]amino]phenyl]ethyl]amino]ethyl]phenyl]formamide;
N-2{2-[4-(3-pheny!-4-methoxyphenyi)aminophenyl]ethyl}-2-hydroxy-2-(8-hydroxy-2(1 H)- quinolinon-5-yl)ethylamine; and
5-[(R)-2-(2-{4-[4-(2-amino-2-methyl-propoxy)-phenylamino]-phenyl}-ethylamino)-1- hydroxy-ethyl]-8-hydroxy-1 H-quinolin-2-one.
The 32-adrenoreceptor agonist may be in the form of a salt formed with a pharmaceutically acceptable acid selected from sulphuric, hydrochloric, fumaric, hydroxynaphthoic (for example 1- or 3-hydroxy-2-naphthoic), cinnamic, substituted cinnamic, triphenylacetic, sulphamic, sulphanilic, naphthaleneacrylic, benzoic, 4-methoxybenzoic, 2- or 4-hydroxybenzoic, 4-chlorobenzoic and 4-phenylbenzo'ic acid.
Suitable anti-inflammatory agents include corticosteroids. Suitable corticosteroids which may be used in combination with the compounds of formula (I) or pharmaceutically acceptable salts thereof are those oral and inhaled corticosteroids and their pro-drugs which have anti-inflammatory activity. Examples include methyl prednisolone, prednisolone, dexamethasone, fluticasone propionate, 6a,9co-difluoro-11 p-hydroxy-16a- methyl-17a-[(4-methyl-1 ,3-thiazole-5-carbonyl)oxy]-3-oxo-androsta-1 ,4-diene-17β- carbothioic acid S-fluoromethy! ester, 6a,9a-difluoro-17a-[(2-furanylcarbonyl)oxy]-1 1 - hydroxy-16a-methyl-3-oxo-androsta-1 ,4-diene-17p-carbothioic acid S-fiuoromethy! ester (fluticasone furoate), 6a,9a-difluoro-1 1 -hydroxy-16a-methyl-3-oxo-17a-propionyloxy- androsta-1 ,4-diene-17p-carbothioic acid S-(2-oxo-tetrahydro-furan-3S-yl) ester, 6α,9α- difluoro-11 -hydroxy-16a-methyl-3-oxo-17a-(2,2,3,3- tetramethycyciopropylcarbonyl)oxy- androsta-1 ,4-diene-17p-carbothioic acid S-cyanomethyl ester and 6a,9a-difluoro-1 i p- hydroxy-16a-methyl-17a-(1-methycyc!opropylcarbonyl)oxy-3-oxo-androsta-1 ,4-diene-17p- carbothioic acid S-fiuoromethyl ester, beclomethasone esters (for example the 17- propionate ester or the 17,21-dipropionate ester), budesonide, flunisolsde, mometasone esters (for example mometasone furoate), triamcinolone acetonide, rofleponide, ciclesonide (16a, 17-[[(R)-cyclohexylmethylenejbis(oxy)]-1 1 β,21 -dihydroxy-pregna-1 ,4- diene-3,20-dione), butixocort propionate, RPR-106541 , and ST-126. Preferred corticosteroids include fluticasone propionate, 6a,9a-difluoro-1 I p-hydroxy-iea-methyl- ^a-t -methyl-l ^-thiazoie-S-carbony oxyj-S-oxo-andro^^
acid S-f!uoromethy! ester, 6 ,9a-difluoro-17a-[(2-furanyicarbonyl)oxy]-1 1 p-hydroxy-16a- methyl-3-oxo-androsta-1 ,4-diene-17p-carbothioic acid S-fluoromethyi ester, 6α,9 - difluoro-1 1 p-hydroxy-16a-methyl-3-oxo-17a-(2,2,3,3- tetramethycyclopropylcarbonyl)oxy- androsta-1 ,4-diene-17p-carbothioic acid S-cyanomethyl ester and 6a,9a-dsfluoro-11 p- hydroxy-16a-methyl-17a-(1-methycyclopropylcarbonyl)oxy-3-oxo-androsta-1 ,4-dtene-17p- carbothioic acid S-fluoromethyi ester. In one embodiment the corticosteroid is 6α,9α- difluoro-17cc-[(2-furanylcarbonyi)oxy]-1 1 p-hydroxy-16 -methyl-3-oxo-androsta-1 ,4-diene- 17p-carbothioic acid S-fiuoromethyl ester.
Examples of corticosteroids may include those described in WO2002/088167, WO2002/100879, WO2002/12265, WO2002/12266, WO2005/005451 , WO2005/005452, WO2006/072599 and WO2006/072600.
Non-steroidal compounds having glucocorticoid agonism that may possess selectivity for transrepression over transactivation and that may be useful in combination therapy include those covered in the following patents: WO03/082827, W098/54159, WO04/005229, WO04/009017, WO04/018429, WO03/104195, WO03/082787, WO03/082280, WO03/059899, WO03/101932, WO02/02565, WO01/16128, WO00/66590, WO03/086294, WO04/026248, WO03/061651 and WO03/08277. Further non-steroidal compounds are covered in: WO2006/000401 , WO2006/000398 and WO2006/015870.
Examples of anti-inf!ammatory agents include non-steroidal anti-inflammatory drugs (NSAID's).
Examples of NSAID's include sodium cromoglycate, nedocromil sodium, phosphodiesterase (PDE) inhibitors (for example, theophylline, PDE4 inhibitors or mixed PDE3/PDE4 inhibitors), leukotriene antagonists, inhibitors of !eukotriene synthesis (for example montelukast), iNOS inhibitors, tryptase and elastase inhibitors, beta-2 integrin antagonists and adenosine receptor agonists or antagonists (e.g. adenosine 2a agonists), cytokine antagonists (for example chemokine antagonists, such as a CCR3 antagonist) or inhibitors of cytokine synthesis, or 5-lipoxygenase inhibitors. An iNOS (inducible nitric oxide synthase inhibitor) is preferably for oral administration. Examples of iNOS inhibitors include those disclosed in WO93/ 3055, WO98/30537, WO02/50021 , W095/34534 and W099/62875. Examples of CCR3 inhibitors include those disclosed in WO02/26722. In one embodiment, the invention provides the use of the compounds of formula (I) in combination with a phosphodiesterase 4 (PDE4) inhibitor, especially in the case of a formulation adapted for inhalation. The PDE4-specific inhibitor useful in this aspect of the invention may be any compound that is known to inhibit the PDE4 enzyme or which is discovered to act as a PDE4 inhibitor, and which are only PDE4 inhibitors, not compounds which inhibit other members of the PDE family, such as PDE3 and PDE5, as well as PDE4.
Compounds include c/s-4-cyano-4-(3-cyclopentyloxy-4-methoxyphenyl)cyclohexan-1 - carboxylic acid, 2-carbomethoxy-4-cyano-4-(3-cyclopropylmethoxy-4- difluoromethoxyphenyl)cyclohexan-1-one and c/'s-[4-cyano-4-(3-cyclopropylmethoxy-4- difluoromethoxyphenyl)cyclohexan-1 -of]. Also, c/s-4-cyano-4-[3-(cyc!opentyloxy)-4- methoxyphenyl]cyclohexane-1 -carboxylic acid (also known as cilomilast) and its salts, esters, pro-drugs or physical forms, which is described in U.S. patent 5,552,438 issued 03 September, 1996; this patent and the compounds it discloses are incorporated herein in full by reference. Other compounds include AWD-12-281 from Elbion (Hofgen, N. et ai. 15th EFMC tnt Symp Med Chem (Sept 6-10, Edinburgh) 1998, Abst P.98; CAS reference No. 247584020-9); a 9-benzy!adenine derivative nominated NCS-613 (INSERM); D-4418 from Chiroscience and Schering-Plough; a benzodiazepine PDE4 inhibitor identified as C!- 1018 (PD-168787) and attributed to Pfizer; a benzodioxole derivative disclosed by Kyowa Hakko in W099/16766; K-34 from Kyowa Hakko; V- 1294A from Napp (Landelis, L.J. et al. Eur Resp J [Annu Cong Eur Resp Soc (Sept 19-23, Geneva) 1998] 1998, 12 (Supp!. 28): Abst P2393); roflumiiast (CAS reference No 162401 -32-3) and a pthalazinone (WO99/47505, the disclosure of which is hereby incorporated by reference) from Byk- Gulden; Pumafentrine, (-)-p-[(4aR*,10£)S*)-9-ethoxy-1 ,2,3,4,4a,10b-hexahydro-8- methoxy-2-methylbenzo[c][1 ,63naphthyridin-6-y!]-NlN-diisopropylbenzamide which is a mixed PDE3/PDE4 inhibitor which has been prepared and published on by Byk-Gulden, now Altana; arofylline under development by Almiral!-Prodesfarma; VM554/UM565 from Vernalis; or T-440 (Tanabe Seiyaku; Fuji, K. et_al. J Pharmacol Exp Ther,1998, 284(1): 162), and T2585.
Further compounds are disclosed in the published international patent application WO04/024728 (Glaxo Group Ltd), WO04/056823 (Glaxo Group Ltd) and WO04/103998 (G!axo Group Ltd) (e.g. Example 399 or 544 disclosed therein), Further compounds are also disclosed in WO2005/058892, WO2005/090348, WO2005/090353, and WO2005/090354, all in the name of Glaxo Group Limited.
Examples of anticholinergic agents are those compounds that act as antagonists at the muscarinic receptors, in particular those compounds which are antagonists of the or M3 receptors, dual antagonists of the Μ,/Ιν^ or M2/M3, receptors or pan-antagonists of the i/M2/ 3 receptors. Exemplary compounds for administration via inhalation include ipratropium (for example, as the bromide, CAS 22254-24-6, sold under the name Atrovent), oxitropium (for example, as the bromide, CAS 30286-75-0) and tiotropium (for example, as the bromide, CAS 136310-93-5, sold under the name Spiriva). Also of interest are revatropate (for example, as the hydrobromide, CAS 262586-79-8) and LAS- 34273 which is disclosed in WO01/041 18. Exemplary compounds for oral administration include pirenzepine (CAS 28797-61-7), darifenacin (CAS 133099-04-4, or CAS 133099- 07-7 for the hydrobromide sold under the name Enablex), oxybutynin (CAS 5633-20-5, sold under the name Ditropan), terodiline (CAS 15793-40-5), tolterodine (CAS 124937-51- 5, or CAS 124937-52-6 for the tartrate, sold under the name Detrol), otiionium (for example, as the bromide, CAS 26095-59-0, sold under the name Spasmomen), trospium chioride (CAS 10405-02-4) and solifenacin (CAS 242478-37-1 , or CAS 242478-38-2 for the succinate aiso known as Y -905 and sold under the name Vesicare).
Additional compounds are disclosed in WO 2005/037280, WO 2005/046586 and WO 2005/104745, incorporated herein by reference. The present combinations include, but are not limited to:
(3-e/?cfo)-3-(2,2-di-2-thienylethenyl)-8,8-dimethyl-8-azoniabicyclo[3.2.1 ]octane iodide;
(3-enc/o)-3-(2-cyano-2,2-diphenylethyl)-8,8-dimethyl-8-azoniabicyclo[3.2.1]octane bromide;
4-[hydroxy(diphenyl)methyl]-1-{2-[(phenylmethyl)oxy]ethyl}-1 -azoniabicyclo[2.2.2]octane bromide; and
(1 ,5S)-3-(2-cyano-2,2-diphenylethyl)-8-methyl-8-{2-[(phenylmethyi)oxy]ethyl}-8- azoniabicyclo[3.2.1 ]octane bromide. Other anticholinergic agents include compounds which are disclosed in US patent application 60/487981 including, for example:
(3-enafo)-3-(2,2-di-2-thienylethenyi)-8,8-dimethyl-8-azoniabicyclo[3.2.1 ]octane bromide; (3-e 90'o)-3-(2!2-diphenylethenyi)-8,8-dimethyl-8-azoniabicycio[3.2.13octane bromide;
(S-ent oJ-S^^-diphenyletheny -S.S-dimethyl-e-azoniabicycloES^. Ijoctane 4- methyibenzenesulfonate;
(3-e/7C/o)-8,8-dimethy!-3-[2-phenyl-2-(2-thienyl)ethenyl]-8-azoniabicyclot3.2.1]octane bromide; and/or
(3-e/?cio)-8,8-dimethyl-3-[2-phenyl-2-(2-pyridinyi)ethenyi]-8-azoniabicyclo[3.2.1]octane bromide.
Further anticholinergic agents include compounds which are disclosed in US patent application 60/511009 including, for example:
(e/7do)-3-(2-methoxy-2,2-di-thiophen-2-yl-ethyl)-8,8-dimethyl-8-azonia- bicyclo[3.2.1 ]octane iodide;
3-((endo)-8-methyl-8-aza-bicyclo[3.2.1 ]oct-3-yl)-2,2-diphenyl-propionitrile;
(enc/o)-8-methyl-3-(2,2J2-trtphenyl-ethyi)-8-aza-bicyclo[3.2.13octane;
3-((e ?c/o)-8-methyl-8-aza-bicyc!o[3.2.1 ]oct-3-yl)-2,2-diphenyl-propionamide;
3-((e/7cfo)-8-methyl-8-aza-bicyclo[3.2.13oct-3-yl)-2,2-diphenyl-propionic acid;
(e/7c o)-3-(2-cyano-2,2-diphenyl-ethyl)-8,8-dimethyl-8-azonia-bicyc!o[3.2.1 ]octane iodide; (e/?c/o)-3-(2-cyano-2,2-diphenyl-ethyl)-8,8-dimethyl-8-azonia-bicyc!o[3.2.1 ]octane bromide;
3-((e ?do)-8-methyl-8-aza-bicyclo[3.2.1 ]oct-3-yl)-2,2-diphenyl-propan-1-ol;
W-benzyi-3-((enc/o)-8-methyl-8-aza-bicyclo[3.2.1 ]oct-3-yl)-2,2-diphenyl-propionamide; (endo)-3-(2-carbamoyl-2,2-diphenyl~ethyl)-8,8-dim^
iodide;
1-benzyi-3-[3-((endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-2,2-diphenyl-propyl]-urea; 1-ethyi-3-[3-{(endo)-8-methyl-8-aza-bicyclo[3,2.1]oct-3-yi)-2,2-diphenyl-propyl]-urea;
A -[3-((endo)-8-methy(-8-aza-bicyclo[3,2.1 ]oct-3-yl)-2,2-diphenyl-propyi3-aceiamide;
/V-[3-((e/?cfo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yl)-2 2-diphenyl-propyi]-benzamide;
3-((endo)-8-methyl-8-aza-bicyclo[3.2.1]oct-3-yi)-2,2-di-thtophen-2-yl-propionitrile;
(endo)-3-(2-cyano-2,2-di-thiophen-2-yl-ethyl)-8,8-dimethyl-8-azonia-bicyclo[3.2.1]octan iodide;
/V-[3-((e 7c o)-8-methyl-8-aza-bicycio[3.2.1 ]oct-3-yl)-2,2-diphenyl-propyl]- benzenesulfonamide;
[3-((enc/o)-8-methyl-8-aza-bicyclo[3.2.1 ]oct-3-yi)-2,2-diphenyi-propyl]-urea;
W-[3-((e/ido)-8-methyi-8-aza-bicyclo[3.2.1]oct-3-yl)-2,2-diphenyi-propyl]- methanesulfonamide; and/or
(endo)-3-{2,2-diphenyl-3-[(1-pheny]-methanoyl)-amino]-propyl}-8,8-dimethyl-8-a bicyclo[3.2.1 ]octane bromide.
Further compounds include:
(enc o)-3-(2-methoxy-212-di-thiophen-2-yl-ethyl)-8)8-dimethy!-8-azonia- bicyclo[3.2.1 ]octane iodide;
(endo)-3-(2-cyano-212-diphenyl-ethyl)-8,8-dimethyl-8-azonia-bicycio[3.2.1]octane iodide; (e/7 /o)-3-(2-cyano-2,2-diphenyl-ethyl)-8,8-dimethyl-8-azonia-bicycio[3.2.1 ]octane bromide; (erioO)-3-(2-carbamoyl-2,2-diphenyi-ethyl)-8l8-dimethyl-8-azonia-bicyclo[3.2.1]octane iodide;
(e/7£/o)-3-(2-cyano-2l2-di-thiophen-2-yl-ethyl)-8,8-dimethy!-8-azonia-bicycio[3.2.1 ]octane iodide; and/or
(e 7Gto)-3-{2,2-diphenyl-3-[(1-phenyl-methanoyl)-amino]-propyl}-8,8-dimethyl-8-azonia- bicyclo[3.2.1 ]octane bromide, In one embodiment the invention provides a combination comprising a compound of formula (i) or a pharmaceutically acceptable salt thereof together with an H1 antagonist, Examples of H1 antagonists include, without limitation, amelexanox, astemizole, azatadine, azelastine, acrivastine, brompheniramine, cetirizine, levocetirizine, efietirizine, chlorpheniramine, clemastine, cyc!izine, carebastine, cyproheptadine, carbinoxamine, descarboethoxyloratadine, doxylamine, dimethindene, ebastine, epinastine, efietirizine, fexofenadine, hydroxyzine, ketottfen, loratadine, levocabastine, mizolastine, mequitazine, mianserin, noberastine, meclizine, norastemtzole, olopatadine, picumast, pyrilamine, promethazine, terfenadine, tripelennamine, temeiastine, trimeprazine and triproiidine, particularly cetirizine, levocetirizine, efletirizine and fexofenadine. In a further embodiment the invention provides a combination comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof together with an H3 antagonist (and/or inverse agonist). Examples of H3 antagonists include, for example, those compounds disclosed in WO2004/035556 and in WO2006/045416. Other histamine receptor antagonists which may be used in combination with the compounds of the present invention include antagonists (and/or inverse agonists) of the H4 receptor, for example, the compounds disclosed in Jablonowski et a/., J. Med. Chem. 46:3957-3960 (2003).
The invention thus provides, in a further aspect, a combination comprising a compound of formula (!) or a pharmaceutically acceptable salt thereof together with a PDE4 inhibitor.
The invention thus provides, in a further aspect, a combination comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof together with a 2-adrenoreceptor agonist.
The invention thus provides, in a further aspect, a combination comprising a compound of formula (!) or a pharmaceutically acceptable salt thereof together with a corticosteroid.
The invention thus provides, in a further aspect, a combination comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof together with a non-steroidal GR agonist. The invention thus provides, in a further aspect, a combination comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof together with an anticholinergic.
The invention thus provides, in a further aspect, a combination comprising a compound of formula (!) or a pharmaceutically acceptable salt thereof together with an antihistamine.
The invention thus provides, in a further aspect, a combination comprising a compound of formula (I) or a pharmaceutically acceptable sa!t thereof together with a PDE4 inhibitor and a
Figure imgf000074_0001
agonist. The invention thus provides, in a further aspect, a combination comprising a compound of formula (!) or a pharmaceutically acceptable salt thereof together with an anticholinergic and a PDE-4 inhibitor. The combinations referred to above may conveniently be presented for use in the form of a pharmaceutical composition and thus pharmaceutical compositions comprising a combination as defined above together with a pharmaceutically acceptable diluent or carrier represent a further aspect of the invention.
The individual compounds of such combinations may be administered either sequentially or simultaneously in separate or combined pharmaceutical formulations. In one embodiment, the individual compounds will be administered simultaneously in a combined pharmaceutical formulation. Appropriate doses of known therapeutic agents will readily be appreciated by those skilled in the art.
The invention thus provides, in a further aspect, a pharmaceutical composition comprising a combination of a compound of formula (I) or a pharmaceutically acceptable salt thereof together with another therapeutically active agent.
The invention thus provides, in a further aspect, a pharmaceutical composition comprising a combination of a compound of formula (I) or a pharmaceutically acceptable salt thereof together with a PDE4 inhibitor.
The invention thus provides, in a further aspect, a pharmaceutical composition comprising a combination of a compound of formula (I) or a pharmaceutically acceptable salt thereof together with a p2-adrenoreceptor agonist. The invention thus provides, in a further aspect, a pharmaceutical composition comprising a combination of a compound of formula (I) or a pharmaceutically acceptable salt thereof together with a corticosteroid.
The invention thus provides, in a further aspect, a pharmaceutical composition comprising a combination of a compound of formula (I) or a pharmaceutically acceptable salt thereof together with a non-steroidal GR agonist.
The invention thus provides, in a further aspect, a pharmaceutical composition comprising a combination of a compound of formula (I) or a pharmaceutically acceptable salt thereof together with an anticholinergic. The invention thus provides, in a further aspect, a pharmaceutical composition comprising a combination of a compound of formula (I) or a pharmaceuticaily acceptable salt thereof together with an antihistamine. The invention thus provides, in a further aspect, a pharmaceutical composition comprising a combination of a compound of formula (i) or a pharmaceutically acceptable salt thereof together with a PDE4 inhibitor and a p2-adrenoreceptor agonist.
The invention thus provides, in a further aspect, a pharmaceutical composition comprising a combination of a compound of formula (f) or a pharmaceuticaily acceptable salt thereof together with an anticholinergic and a PDE4 inhibitor.
The invention will now be illustrated by way of the following non-limiting examples.
EXAMPLES
The following examples illustrate the invention. These examples are not intended to limit the scope of the present invention, but rather to provide guidance to the skilled artisan to prepare and use the compounds, compositions, and methods of the present invention. While particular embodiments of the present invention are described, the skilled artisan wil! appreciate that various changes and modifications can be made without departing from the spirit and scope of the invention. General Methods
Unless stated otherwise, starting materials were commercially available. All solvents and commercial reagents were of laboratory grade and were used as received. Unless stated otherwise, flash chromatography was carried out using pre-packed Biotage "isolute" flash silica cartridges on a Biotage "Flashmaster 2" system.
The following methods were used for LCMS (liquid chromatography - mass spectral) analysis:
LCMS Method A;
The analysis was conducted on an Acquity UPLC BEH C18 column (50mm x 2.1 mm internal diameter 1 .7pm packing diameter) at 40°C.
The solvents employed were:
A = 0.1 % v/v solution of formic acid in water.
B = 0.1 % v/v solution of formic acid in acetonitrile.
The gradient employed was as follows:
Time Flow Rate
% A % B
(minutes) (mL/min)
0 1 97 3
1.5 1 0 100
Figure imgf000078_0001
The UV detection was an averaged signal from wavelength of 210nm to 350nm and mass spectra were recorded on a mass spectrometer using alternate-scan positive and negative mode electrospray ionization.
LCMS Method B:
The analysis was conducted on an XBridge C18 column {50mm x 4.6mm interna! diameter 3.5μηι packing diameter) at 30°C.
The solvents employed were:
A = 10 mM ammonium bicarbonate in water adjusted to pH 10 with ammonia solution. B = acetonitrile.
The typical gradient employed was as follows:
Figure imgf000078_0002
The UV detection was an averaged signal from wavelength of 210nm to 350nm and mass spectra were recorded on a mass spectrometer using alternate-scan positive and negative mode electrospray ionization.
LCMS Method C: The analysis was conducted on an a Sunfire C18 column (30mm x 4.6mm i.d. 3.5pm packing diameter) at 30°C.
The solvents employed were: A = 0.1 % v/v solution of formic acid in water.
B = 0.1 % v/v solution of formic acid in acetonitriSe. The gradient employed was as follows:
Figure imgf000079_0001
The UV detection was an averaged signal from wavelength of 210nm to 350nm and mass spectra were recorded on a mass spectrometer using alternate-scan positive and negative mode electrospray ionization.
LCMS Method D:
The analysis was conducted on an Acquity UPLC BEH C18 column (50mm x 2.1 mm internal diameter 1.7pm packing diameter) at 40°C.
The solvents employed were:
A = 10 mM ammonium bicarbonate in water adjusted to pH 10 with ammonia solution. B = acetonitrile.
The gradient employed was as follows:
Time Flow Rate
% A % B
(minutes) (mL/min)
0 1 99 1
1.5 1 3 97
1.9 1 3 97 2.0 1 0 100
The UV detection was an averaged signal from wavelength of 210nm to 350nm and mass spectra were recorded on a mass spectrometer using alternate-scan positive and negative mode electrospray ionization.
The following illustrates the mobile phases and gradients used when compounds underwent purification by mass-directed autopreparative HPLC. LCMS Method E:
The analysis was conducted on an Acquity UPLC BEH C18 column (50mm x 2.1 mm internal diameter 1.7pm packing diameter) at 40°C. The solvents employed were:
A = 0.1 % v/v solution of Trifiuoroacetic Acid in Water.
B = 0.1 % v/v solution of Trifiuoroacetic Acid in Acetonitrile
The gradient employed was as follows
Figure imgf000080_0001
The UV detection was an averaged signal from waveiength of 210nm to 350nm and spectra were recorded on a mass spectrometer using alternate-scan positive and negative mode electrospray ionization.
Mass-Directed Autopreparative HPLC (Formic Acid Modifier) The HPLC analysis was conducted on a Sunfire C18 column (150mm x 30mm internal diameter, 5pm packing diameter) at ambient temperature.
The solvents employed were:
A = 0.1 % v/v solution of formic acid in water.
B = 0.1 % v/v solution of formic acid in acetonitrile.
Mass-Directed Autopreparative HPLC (Trifluoroacetic Acid Modifier)
The HPLC analysis was conducted on a Sunfire C18 column (150mm x 30mm interna! diameter, 5pm packing diameter) at ambient temperature.
The solvents employed were:
A = 0.1 % v/v solution of trifluoroacetic acid in water.
B = 0.1 % v/v solution of trifluoroacetic acid in acetonitrile.
Mass-Directed Autopreparative HPLC (Ammonium Bicarbonate Modifier)
The HPLC analysis was conducted on an XBridge C18 column (150mm x 30mm internal diameter, 5pm packing diameter) at ambient temperature.
The solvents employed were:
A = 10 mM ammonium bicarbonate in water adjusted to pH 10 with ammonia solution. B = acetonitrile.
For each of the mass-directed autopreparative purifications, irrespective of the modifier used, the gradient employed was dependent upon the retention time of the particular compound undergoing purification as recorded in the analytical LCMS, and was as follows: For compounds with an analytical LCMS retention time below 0.6 minutes (LCMS method A) or below 1.5 minutes (LCMS method B) the following gradient was used: Time Flow Rate
% A % B
(minutes) (mL/min)
0 40 99 1
1 40 99 1
10 40 70 30
11 40 1 99
15 40 1 99
For compounds with an analytical LCMS retention time between 0.6 and 0.9 minutes (LCMS method A) or between 1.5 and 2.2 minutes (LCMS method B) the following gradient was used:
Figure imgf000082_0001
For compounds with an analytical LCMS retention time between 0.9 and 1 ,2 minutes (LCMS method A) or between 2.2 and 3.0 minutes (LCMS method B) the following gradient was used:
Figure imgf000082_0002
For compounds with an analytical LCMS retention time between 1.2 and 1 .4 minutes (LCMS method A) or between 3.0 and 3.6 minutes (LCMS method B) the following gradient was used: Time Flow Rate
% A % B
(minutes) (mL/min)
0 40 50 50
1 40 50 50
10 40 1 99
1 1 40 1 99
15 40 1 99
For compounds with an analytical LC S retention time greater than 1.4 minutes (LCMS method A) or greater than 3.6 minutes (LCMS method B) the following gradient was used:
Figure imgf000083_0002
The UV detection was an averaged signa! from wavelength of 210nm to 350nm and mass spectra were recorded on a mass spectrometer using alternate-scan positive and negative mode electrospray ionization.
The chemical names were generated using ACD Name Pro version 6.02 from Advanced Chemistry Development, Inc.
Intermediate 1 :
-[{methyloxy)methyl]-2-(methylthio)-4-pyrimidinyi trifluoromethanesulfonate
Figure imgf000083_0001
A mixture of 6-[(methyloxy)methyl]-2-(methylthto)-4(1 H)-pyrimidinone (5 g, 26.8 mmoi) and W,A/-diisopropylethyiamine (1 1.72 mL, 67.1 mmoi) in dichloromethane (20 mL) was stirred at 0 °C. Trifiic anhydride (6.68 mL, 40.3 mmoi) was then added dropwise. The reaction mixture was brought to room temperature and stirred for 1 hour. Water was added and the organic layer was dried over magnesium sulfate, filtered and evaporated to dryness. The oil was purified by chromatography on silica using a gradient elution from 0 to 50 % ethyl acetate in cyclohexane to afford the title compound (7.19 g, 22.59 mmol, 84 % yield) as a light brown oil LCMS (Method A): Rt 1.21 minutes; m/z 319 (MH+).
Intermediate 2:
-[(methyloxy)methyl]-2-(methylthio)-4-pyrimidinamine
Figure imgf000084_0001
To a solution of 6-[(methyloxy)methyl]-2-(methylthio)-4-pyrimidinyl trifluoromethanesulfonate (46.7 g, 147 mmol) in anhydrous acetonitrile (200 mL) was added W./V-diisopropylethylamine (36.4 mL, 220 mmo!) and terf-octylamine (36.7 mL, 220 mmol). The reaction mixture was stirred at reflux for 7 hours. The oil was taken up in dichloromethane (300 mL) and water (300 mL) then washed with saturated sodium bicarbonate solution (200 mL). The organic material was isolated, dried over magnesium sulfate and evaporated to dryness to give a brown oil. Trifluoroacetic acid (200 mL, 2596 mmol) was added and the mixture was stirred at room temperature under nitrogen for 2 hours. The reaction was heated at 50 °C under nitrogen for 3 hours. The reaction mixture was concentrated to dryness to afford a brown oil. The oil was taken up in dichloromethane (100 mL), neutralised with saturated sodium bicarbonate solution (200 mL) followed by solid sodium bicarbonate. Once neutral, the mixture was diluted with water (200 mL) and the organic phase was separated. The aqueous phase was extracted with dichloromethane (3 x 200 mL). The organic phases were dried over magnesium sulfate and evaporated to dryness to give a light brown solid. The solid was dissolved in dichloromethane (100 mL) and passed through an aminopropyl column, eluting with dichloromethane (200 mL). The desired fractions were combined and concentrated to dryness to afford the title compound (23.4 g, 126 mmol, 86 % yield) as a light brown solid. LCMS (Method B): Rt 1.60 minutes; m/z 186 (MH+). Intermediate 3:
W-[6-[(methyloxy)methyl]-2-(methylthio)-4-pyri
amine
Figure imgf000085_0001
A solution of 6-[(methyloxy)methy!]-2-(methylthio)-4-pyrimidinamine (10 g, 54 mmol) in /V,W-dimethylformamide (100 mL) was cooled down to 0 °C and sodium hydride (60 % w/w in mineral oil) (3.24 g, 81 mmol) was carefully added under nitrogen. The reaction was stirred for 20 minutes until the effervescence had stopped. 2-bromo[1 ,3]thiazolo[5,4- 0]pyridine (1 1.61 g, 54 mmol) was added and the reaction mixture was stirred at room temperature overnight. The reaction mixture was cooled to 0 °C, sodium hydride (60 % w/w in mineral oil) (1.08 g, 27 mmol) was added and the reaction slowly warmed up to 40 °C and stirred for 4 hours. Aqueous ammonium chloride was added and the reaction was stirred for 10 minutes. The mixture was poured into water and the light brown precipitate was then filtered off, dried overnight under reduced pressure in the oven to afford the title compound (18.4 g, 57.6 mmol, >100 % yield) as a light brown solid. LCMS (Method A): Rt 0.90 minutes; m/z 320 (MH+). Intermediate 4:
W-[6-[(methyloxy)methyl]-2-(methylsulfony0
/> ridin-2-amine
Figure imgf000085_0002
To a stirred solution of A/-[6-[(methyloxy)methyl]-2-(methylthio)-4- pyrimidinyl][1 ,3]thiazolo[5,4-jb]pyridin-2-amine (18.4 g, 57.6 mmol) in N,N- dimethylformamide (100 mL) was added oxone® (70.8 g, 1 15 mmol) portionwise and the reaction was stirred at room temperature for 4 hours. Sodium bicarbonate was added and the reaction was stirred for 20 minutes. The mixture was poured into 300 mL of dichloromethane and the cream solid was filtered off, dried under reduced pressure in the oven to give 36.8 g of a beige solid. 21 g of the solid was suspended in dichloromethane: 2M ammonia in methanol (1 : 1 , 400 mL) and placed in the ultrasonic bath. The solution was fiitered and washed twice with dichloromethane:2M ammonia in methanol (1 :1 ). The fiitrate was evaporated to dryness to give title compound (5.5 g, 15.67 mmol, 27 % yield) as yellow solid. LCMS (Method B): Rt 1.51 minutes; m/z 352 (MH+). Intermediate 5:
1 ,1-dimethyiethyl (2-{[4-[(methy!oxy)methy!3-6-([1,3]thiazolo[5,4-i)]pyridin-2- yfamino}-2-pyrimsdinyl]amino}ethyl)carbamate
Figure imgf000086_0001
A microwave vial was charged with A/-[6-[(methyloxy)methyl]-2-(methylsulfonyl)-4- pyrimidinyl][1 ,3]thiazolo[5,4-0]pyridin-2-amine (450 mg, 1.281 mmol) and 1 ,1 - dimethylethyl (2-aminoethyl)carbamate (308 mg, 1.921 mmol) in isopropanol (10 mL). The via! was sealed and was heated in a Biotage Initiator microwave system at 150 °C for 30 minutes. The residue was then evaporated to dryness. The product was purified by chromatography on silica using a gradient elution from 0 to 30 % methanol (+1 % triethylamine) in dichloromethane to afford the title compound (504 mg, 1.168 mmol, 91 % yield) as an off-white solid. LC S: (Method B): Rt 2.37 minutes; m/z 432 (MH+).
Intermediate 6:
Λ/^ίΣ-ΗηΊίηοθΐ ν -β-ΠΓηβ^ΙοχνΪΓηθ^νΙΙ- ί4-!! ,3]thiazolo[5,4- >]pyridin-2-yl-2,4- pyrimidinediamine hydrochloride
Figure imgf000086_0002
A mixture of 1 , 1 -dimethylethyl (2-{[4-[(methyloxy)methyl]-6-([1 ,3]thiazolo[5,4-b]pyridin-2- yiamino)-2-pyrimidinyl]amino}ethyl)carbamate (500 mg, 1.159 mmol) and 4M HCI in dioxane (1.45 mL, 5.79 mmol) in dichloromethane (10 mL) was stirred at room temperature for 16 hours. 4 HCI in dioxane (1.45 mL, 5.79 mmol) was added and the reaction mixture stirred at room temperature for 3 hours. The solid was filtered under reduced pressure and washed with dichloromethane (2 x 10 mL). The white solid was dried under reduced pressure to give the title compound (400 mg, 1.087 mmol, 94 % yield). LCMS: (Method C): Rt 1.04 minutes; m/z 332 (MH+).
In a second preparation, the free base of the title compound was synthesised. Intermediate 7:
6-bromo-W-(2-ch]oro-4-pyrimidinyl)-1,3-benzothiazoi-2-amine
Figure imgf000087_0001
To a solution of 2-chloro-4-pyrimidinamine (1 g, 7.72 mmol) in anhydrous tetrahydrofuran (5 mL) at 0 °C was added sodium hydride (60% w/w in mineral oil) (0.926 g, 23.16 mmol) portionwise. The reaction mixture was stirred for 10 minutes at 0 °C under nitrogen atmosphere then 6-bromo-2-chloro-1 ,3-benzothiazole (2.11 g, 8.49 mmol) was added portionwise. The reaction mixture was stirred at 0 °C under nitrogen atmosphere for 15 minutes, stirred allowing to warm up to room temperature and then heated to 65 °C overnight under nitrogen atmosphere. The reaction mixture was allowed to cool to room temperature then water (30 mL) was added and the mixture extracted with ethyl acetate (2 x 50 mL). The aqueous phase was filtered to give the title compound (393 mg, 1.150 mmol, 15 % yield) as a cream solid. LCMS: (Method C): Rt 3.06 minutes; m/z 341 , 343 (MH+).
Intermediate 8:
W2-(2-aminoethyl)-W4-(6-bromo-1 ,3-benzothiazoi-2-yl)-2,4-pyrimidinediamine
Figure imgf000087_0002
A microwave vial was charged with 6-bromo-A/-(2-chioro-4-pyrimidinyl)-1 ,3-benzothiazol- 2-amine (281 mg, 0.823 mmol), 1 ,2-ethanediamine (0.165 mL, 2.468 mmol) and isopropanol (4 mL). The microwave vial was sealed and was heated in a Biotage Initiator microwave to 150 °C for 45 minutes. After cooling, the solid was filtered under reduced pressure to give a yellow solid. The solid was transferred into a microwave vial. 1 ,2- Ethanediamine (0.165 mL, 2.468 mmol) and isopropanol (4 mL) were added. The vial was sealed and was heated in a Biotage Initiator microwave to 150 °C for 20 minutes. After cooling the reaction mixture was filtered under reduced pressure to give the title compound (194 mg, 0.531 mmol, 64% yield) as a yellow solid. LCMS: (Method B): Rt 2.19 minutes; m/z 365, 367 (MH+). Intermediate 9:
1 ,1-dimethylethyl 4-{[4-[(methyloxy)methyl]-6-([1 ,3]thiazolo[5,4-b3pyridin-2-ylamino}- -pyrimidinyl]amino}-1-piperidinecarboxylate
A microwave vial was charged with A/-[6-[(methyloxy)methyl]-2-(methylsulfonyl)-4- pyrimidinyl][1 ,3]thia2olo[5,4-0]pyridin-2-amine (500 mg, 1.423 mmol), 1 ,1-dimethylethyl 4- amtno-1-piperidinecarboxylate (427 mg, 2.134 mmol) and isopropanol (5 mL). The vial was sealed and was heated in a Biotage Initiator microwave at 150 °C for 45 minutes. After cooling, the reaction was filtered under reduced pressure to give a yellow solid. The solid was purified by chromatography on silica using a gradient elution from 0 to 25 % methanol in dichloromethane to afford the title compound (287 mg, 0.608 mmol, 43 % yield). LCMS: (Method A): Rt 0.84 minutes; m/z 472 (MH+).
Intermediate 10:
6-[(methyloxy)methyl]-W2-4-pipendinyl^
pyrimidinediamine hydrochloride
Figure imgf000088_0002
1 ,1-Dimethylethyl 4-{[4-t(methyloxy)methyl]-6-([1 ,33thiazolo[5,4-i)]pyndin-2-ylamino)-2- pyrimidinyl]amino}-1-piperidinecarboxylate (237 mg, 0.503 mmol) was dissolved in dichloromethane and treated with 4M HCI in dioxane (0.6 mL, 2.4 mmol) at room temperature. The reaction mixture was stirred at room temperature overnight. The reaction mixture was filtered under reduced pressure and the residue was dried to afford the title compound (200 mg, 0.491 mmol, 97 % yield). LCMS (Method C): Rt 1.05 minutes; m/z 372 (MH+). intermediate 11 :
frans-4-{[4-[(methyloxy)methyl]-6-([1,3]thiazolo[5,4-iE>]pyridin-2-ylamino)-2- pyrimidinyl]amino}cyclohexanol
Figure imgf000089_0001
A microwave vial was charged with /V-[6-[(methyioxy)methyl]-2-(methylsulfonyl)-4- pyrimidinyi][1 ,3]thiazolo[5,4-b]pyridin-2-amine (250 mg, 0.71 1 mmol), trans-4- aminocyclohexanol (246 mg, 2.134 mmol) and isopropanol (5 mL). The vial was sealed and was heated in a Biotage Initiator microwave to 150 °C for 45 minutes. After cooling, the reaction was filtered under reduced pressure and the solid was washed with isopropanol. The filtrate was evaporated to dryness. The residue was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (36 mg, 0.093 mmol, 13 % yield). LCMS (Method D): t 0.75 minutes; m/z 387 (MH+).
Intermediate 12:
1 ,1 -dimethylethyl (4-{[4-t(methyloxy)methy!]-6-([1 ,33thiazolo[5,4-*)]pyndin-2- ylamino)-2-pyrimidinyi3amino}butyl)carbamate
Figure imgf000089_0002
A microwave via! was charged with W-[6-[(methyloxy)methyl]-2-(methyisulfonyl)-4- pyrimidinyl][1 ,3]thiazolo[5,4-j ]pyhdin-2-amine (150 mg, 0.427 mmol), 1 , 1 -dimethylethyl (4- aminobutyl)carbamate (0.245 mL, 1.281 mmol) and isopropanol (4 mL). The vial was sealed and was heated in a Biotage Initiator microwave at 150 °C for 45 minutes. After cooling, the reaction mixture was filtered under reduced pressure and the solid was washed with isopropanol. The filtrate was evaporated to dryness to give a white solid. The solid was purified by chromatography on silica using a gradient elution from 0 to 30 % methanol (+1 % triethylamine) in dichloromethane to afford the title compound (146 mg, 0.318 mmol, 74 % yield). LCMS: (Method C): Rt 1.79 minutes; m/z 460 (MH+).
Intermediate 13: W2-(4-aminobutyl}-6-[(methyloxy)meth^^
pyrimtdinediamine hydrochloride
Figure imgf000090_0001
1 , 1 -dimethylethyi (4-{[4-[(meihyioxy)methyl]-6-(E1 ,3]thiazolo[5,4-b3pyridin-2-yiamino)-2- pyrimidinyl]amino}butyl)carbamate (146 mg, 0.318 mmol) was dissolved in dichioromethane (4 mL) and methanol (1 mL) and treated with 4M HCI in dioxane (0.318 mL, 1.271 mmol). The reaction mixture was stirred at room temperature overnight. The reaction mixture was evaporated to dryness to afford the title compound (144 mg, 0.364 mmo!, >100 % yield). LCMS: (Method C): Rt 1.00 minutes; m/z 360 (MH+).
Intermediate 14:
1 ,1 -dimethylethyi (3-{[4-[(methyloxy)methyl3-6-([1 ,3]thiazolo[5,4-/>]pyridin-2- ylamino)-2-pyrimidinyl]amino}propyl)carbamate
Figure imgf000090_0002
A microwave vial was charged with A/-[6-[(methyloxy)methyl3-2-(methylsulfonyl)-4- pyrimidinyl3[1 ,3]thiazolo[5,4-d]pyridin-2-amine (150 mg, 0.427 mmol), 1 , 1 -dimethylethyi (3- aminopropyi)carbamate (0,224 mL, 1.281 mmol) and isopropanol (4 mL). The vial was sealed and was heated in a Biotage Initiator microwave to 150 °C for 45 minutes. After cooling, the reaction was filtered under vacuum and the solid was washed with isopropanol to give the title compound (86 mg).
The filtrate was evaporated to dryness to give a yeilow solid. The solid was purified by chromatography on silica using a gradient elution from 0 to 30% methanol (+1 % triethylamine) in dichloromethane to afford the title compound (52 mg). Both batches of the product were combined to give the title compound (138 mg, 0.31 mmol, 72 % yield). LCMS: (Method B): Rt 2.48 minutes; m/z 446 (MH+). Intermediate 15:
Wz-(3-aminopropyl)-6-[(methyloxy)methyi]-A^
pyrimidinediamine hydrochloride
Figure imgf000091_0001
1 ,1-dimethylethyI (3-{[4-[(methyloxy)methy]]-6-(E1 ,3]thiazo!o[5,4-i)]pyridin-2-ylamino)-2- pyrimidinyl]amino}propyi)carbamate (1 15 mg, 0.258 mmo!) was dissolved in dichloromethane (4 mL) and methanol (1 mL) and treated with 4M HCI in dioxane (0.4 mL, 1.6 mmoi). The reaction mixture was stirred at room temperature overnight, The reaction mixture was evaporated to dryness to afford the title compound (148 mg, 0.388 mmoi, >100 % yield). LCMS: (Method C): Rt 0.99 minutes; m/z 346 (MH+).
Intermediate 16:
W^t ans^-aminocyclohexyll-G-ftmethyloxyimethy^
-2,4-pyrimidinediamine
Figure imgf000091_0002
A microwave via! was charged with /V-[6-[(methyloxy)methyl]-2-(methylsulfonyl)-4- pyrimidinyl3[1 ,3]thiazolo[5,4-0]pyridin-2-amine (200 mg, 0.569 mmol), rrans-1 ,2- cyclohexanediamine (97 mg, 0.854 mmol) and isopropanol (4 mL). The via! was sealed and was heated in a Biotage Initiator microwave at 150 °C for 30 minutes. The mixture was evaporated to dryness to give a yeliow solid. The solid was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (1 17 mg, 0.303 mmol, 53 % yield). LCMS (Method B): Rt 2.07 minutes; m/z 386 (MH+). Intermediate 17:
W2-[c s-3-aminocyclohexyl3-6-[(methyioxy)methyl]-/^-[1 ,3]thiazolo[5,4-J ]pyridin-2-yl- 2,4-pyrimidinediamine
Figure imgf000092_0001
A microwave vial was charged with A/-[6-[(methyloxy)methyi]-2-(methylsulfony!)-4- pyrimidinyl][1 ,3]thiazolo[5,4-j ]pyridin-2-amine (200 mg, 0.569 mmol), 1 ,3- cyclohexanediamine (97 mg, 0.854 mmol) and isopropanol (4 mL). The vial was sealed and was heated in a Biotage Initiator microwave at 150 °C for 30 minutes. After cooling the reaction was filtered under reduced pressure and the solid was washed with isopropanol. The filtrate was evaporated to dryness to give a yeliow solid. The solid was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (73 mg, 0.189 mmol, 33 % yield). LCMS {Method B): Rt 1.91 minutes; m/z 386 (MH+).
Intermediate 18:
1 ,1 -dimethylethyl (3S)-3-{[4-[(methyloxy)methyl3-6-([1 )33thiazolo[5l4-/)]pyridin-2- ylamino)-2-pyrirnidinyl]amino}-1-pyrrolidinecarboxylate
Figure imgf000092_0002
A microwave vial was charged with A/-[6-[(methyloxy)methyl]-2-(methylsu!fonyl)-4- pyrimidinyl][1 ,3]thiazolo[5,4-6]pyridin-2-amine (200 mg, 0.569 mmol), 1 , 1 -dimethylethyl (3S)-3-amino-1-pyrrolidinecarboxylate (159 mg, 0.854 mmol) and isopropanol (4 mL). The vial was sealed and was heated in a Biotage Initiator microwave at 150 °C for 30 minutes. 1 ,1-dimethylethyl (3S)-3-amino-1 -pyrrolidinecarboxylate (159 mg, 0.854 mmol) was added and the vial was heated in a Biotage Initiator microwave at 150 °C for 30 minutes. The soiution was evaporated to dryness to give a yellow solid. The solid was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (86 mg, 0.188 mmol, 33 % yield). LCMS (Method D): Rt 1.05 minutes; m/z 458 (MH+).
Intermediate 19: 6-[(methyloxy)methy^W2-[(3S)-3-py^
pyrirnidinediamine hydrochloride
Figure imgf000093_0001
1 ,1-Dimethylethyl (3S)-3-{[4-[(meihyloxy)meihyl]-6-([1 ,3]thiazolo[5,4-jb]pyridin-2-ylarnino)- 2-pyrimidinyl]amino}-1-pyrrolidinecarboxylate (80 mg, 0.175 mmo!) was dissolved in dichloromethane (3 mL) and treated with 4M HCI in dioxane (0.262 mi_, 1.049 mmol). The reaction was stirred at room temperature overnight. The mixture was evaporated to dryness to afford the title compound (84 mg, 0.213 mmol, >100 % yield). LCMS (Method B): Rt 1.73 minutes; m/z 358 (MH+).
Intermediate 20:
1 ,1 -dimethylethyl 4-[4-[(methyloxy)methyl]-6-{[1 ,3]thiazolo[5,4-&]pyridin-2-ylamino)- -pyrimidinyl]-1 -piperazinecarboxylate
Figure imgf000093_0002
A microwave vial was charged with A/-[6-[(methyloxy)methyl]-2-(methylsulfonyi)-4- pyrimidinyl][1 ,3]thiazolo[5,4-i)]pyridin-2-amine (200 mg, 0.569 mmo!), 1 ,1 -dimethylethyl 1 - piperazinecarboxy!ate (159 mg, 0.854 mmol) and isopropanoi (4 mL). The via! was sealed and was heated in a Biotage Initiator microwave at 150 °C for 30 minutes. A further portion of 1 , 1 -dimethylethyl 1-piperazinecarboxylate (159 mg, 0.854 mmol) was added and the vial was heated at 150 °C for 30 minutes. The solution was evaporated to dryness to give a yellow solid. The solid was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (72 mg, 0.157 mmol, 28 % yield). LCMS (Method B): Rt 3.01 minutes; m/z 458 (MH+). Intermediate 21 :
W-[6-[(methyloxy)methyl]-2-(1-piperazinyl)-4-pyri
2-amine hydrochloride
Figure imgf000094_0001
1 , 1 -Dimethylethyl 4-[4-[(methyloxy)methyi3-6-([1 ,3]thiazoio[5,4-fa]pyridin-2-ylamino)-2- pyrimidinyi]-1-piperazinecarboxylate (67 mg, 0.146 mmoi) was dissolved in dichloromethane (4 ml_) and treated with 4M HCi in dioxane (0.227 mL, 0.908 mmol). The reaction mixture was stirred at room temperature overnight. The mixture was evaporated to dryness to afford the title compound (57 mg, 0.145 mmol, 99 % yield). LCMS (Method B): Rt 1.91 minutes; m/z 358 (MH+).
Intermediate 22:
W2-[c/'s-2-aminocyc!ohexyl]-6-[(methyloxy)m
-pyrimidinediamine
Figure imgf000094_0002
A microwave vial was charged with A/-[6-[(methyloxy)methyl]-2-(methylsulfonyl)-4- pyrimtdinyljt Sjthiazo!oiS^-^pyridin^-amine (200 mg, 0.569 mmoi), c/s-1 ,2- cyclohexanediamine (97 mg, 0.854 mmol) and isopropano! (4 mL). The vial was sealed and was heated in a Biotage Initiator microwave at 150 °C for 30 minutes. The mixture was evaporated to dryness to give a ye!low solid. The solid was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (124 mg, 0.322 mmoi, 56 % yield). LCMS (Method B): Rt 2.11 minutes; m/z 386 (MH+).
Intermediate 23:
W2-(frans-4-aminocyclohexyl)-6-[(methyloxy)m
yl-2,4-pyrimidinediamine
Figure imgf000094_0003
A microwave vial was charged with /V-[6-[(methyloxy)methyl]-2-(methy!sulfonyl)-4- pyrimidinyl][1 ,3]thiazolo[5,4-0]pyridin-2-amine (200 mg, 0.569 mmol), trans-" ,A- cyclohexanediamine (65 mg, 0.569 mmol) and isopropano! (4 mL). The vial was sealed and was heated in a Biotage initiator microwave at 50 °C for 30 minutes. After cooling, the reaction was filtered under reduced pressure and the solid was washed with isopropanol. The filtrate was evaporated to dryness to give a yellow solid. The solid was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (93 mg, 0.241 mmol, 42 % yield). LCMS (Method C): Rt 1.02 minutes; m/z 386 ( H+).
Intermediate 24:
1 ,1-dimethylethyl (3R)-3-{[4-[(rnethyloxy}rnethyU-6-{[1 ,3]thiazolo[5,4-Jb3pyridin-2- ylamino)-2-pyrimidinyl]amino}-1-pyrrolidinecarboxylate
Figure imgf000095_0001
A microwave vial was charged with A/-[6-[(methyloxy)methy!]-2~(methylsulfonyi)-4- pyrimidinyl][1 ,3]thiazolo[5,4-i>3pyridin-2-amine (200 mg, 0.569 mmol), 1 , 1-dimethylethyl (3R)-3-amino-1-pyrrolidinecarboxylate (159 mg, 0.854 mmol) and isopropanol (4 mL). The vial was sealed and was heated in a Biotage Initiator microwave at 150 °C for 30 minutes. A further portion of 1 ,1-dimethylethy! (3R)-3-amino-1-pyrrolidinecarboxyiate (159 mg, 0.854 mmol) was added and the vial was heated in a Biotage initiator microwave at 150 °C for 30 minutes. The mixture was evaporated to dryness to give a yellow solid. The solid was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (80 mg, 0.175 mmol, 31 % yield). LCMS (Method D): Rt 1.05 minutes; m/z 458 (MH+).
Intermediate 25:
6-[{methyloxy)methyl]-W2-[(3R)-3-pyrrolid^
pyrimidinediamine hydrochloride
Figure imgf000096_0001
1 , 1 -Dimethylethyl (3R)-3-{[4-[(methyloxy)methyi]-6-([1 ,3]thiazolo[5,4-jb]pyridsn-2-ylamino)- 2-pyrimidinyl]amino}-1 -pyrrolidinecarboxylate (75 mg, 0.164 mmol) was dissolved in dichloromethane (3 mL) and treated with 4M HCi in dioxane (0.5 ml_, 2 mmol). The reaction mixture was stirred at room temperature overnight. The mixture was evaporated to dryness to afford the title compound (78 mg, 0.198 mmol, >100 % yield). LCMS (Method B): Rt 1.73 minutes; m/z 358 (MH+).
Intermediate 26:
W-E6-[(methyloxy)methyl]-2-(4-piperidinyloxy)-4-pyrimidiny!][1 ,3]th
drochloride
Figure imgf000096_0002
A microwave vial was charged with A/-[6-[(methyioxy)methyl]-2-(methylsulfony!)-4- pyrimidinyl][1 ,3]thiazolo[5,4-t>]pyridin-2-amine (200 mg, 0.569 mmol), 1 ,1 -dimethylethyl 4- hydroxy-1-piperidinecarboxylate (229 mg, 1.138 mmol) and sodium hydride (60 % w/w in mineral oil) (68.3 mg, 1.707 mmol) in tetrahydrofuran (3 ml). The vial was sealed and was heated in a Biotage Initiator microwave at 130 °C for 30 minutes. The reaction mixture was partitioned between dichloromethane and water. The organic layer was dried using a hydrophobic frit and evaporated to dryness to give the required intermediate. The intermediate was dissolved in dichloromethane (3 mL) and methanol (1 ml) and was treated with 4M HCI in dioxane (0.71 1 mL, 2.85 mmol). The reaction was stirred at room temperature overnight. The reaction mixture was evaporated to dryness to afford the title compound (160 mg, 0.391 mmol, 69 % yield). LCMS (Method B): Rt 1.69 minutes; m/z 373 (MH+).
Intermediate 27:
methyl 2-chloro-6-[(1,1 ,3t3-tetramethylbutyl)amino]-4-pyrimidinecarboxylate
Figure imgf000097_0001
A mixture of methyl 2,6-dichloro-4-pyrimidinecarboxylate (10 g, 48.3 mmol), tert- octyiamine (1 1.63 mL, 72.5 mmol) and /V, v"-diisopropylethylamine (12.66 mL, 72.5 mmol) in tetrahydrofuran (10 mL) was stirred at room temperature for 96 hours. The reaction mixture was evaporated to dryness. The residue was partitioned between dichioromethane (200 mL) and saturated aqueous NaHC03 (150 mL). After separation, the organic extract was dried using a hydrophobic frit and evaporated to dryness. The residue was purified by chromatography on silica using a gradient elution from 0 to 60 % ethyl acetate in cyc!ohexane to afford the title compound (10.01 g, 33.4 mmol, 69 % yield) as a white solid. LCMS (Method B): Rt 3.32 minutes; m/z 300 (MH+).
Intermediate 28:
2-chloro-6-(4-morpholinylcarbonyl)-W-(1 ,1 ,3,3-tetramethylbutyl)-4-pyrimidin
Figure imgf000097_0002
A mixture of methyl 2-chloro-6-[(1 , 1 ,3,3-tetramethyibutyi)amino]-4-pyrimidinecarboxylate (5 g, 16.68 mmol), 1 , 3,4,6, 7,8-hexahydro-2H-pyrimido[1 ,2-a]pyrimidine (0.696 g, 5 mmol) and morpholine (1.453 mL, 16.68 mmol) in tetrahydrofuran (30 mL) was stirred at room temperature for 3 hours. The reaction mixture was partitioned between dichioromethane (100 mL) and saturated aqueous Na2C03 (100 mL). After separation, the organic extract was dried using a hydrophobic frit and evaporated to dryness to afford the title compound (5.9 g, 16.62 mmol, 100 % yield) as a white soiid. LCMS (Method B): Rt 3.07 minutes; m/z 355 (MH+).
Intermediate 29:
2-chloro-6-(4-morpholinylmethyl)-/V-{1 )1 ,3,3-tetramethylbutyl)-4-pyrimidinamine
Figure imgf000097_0003
1 Borane in tetrahydrofuran (49.9 mL, 49.9 mmol) was added dropwise to a solution of 2~chloro-6-(4-morphoiinylcarbony!)-A/-(1 , 1 l3,3-tetramethylbutyl)-4-pyhmidinamine (5.9 g, 16.63 mmo!) stirring at 0 °C in tetrahydrofuran (25 mL) under nitrogen atmosphere. The reaction mixture was stirred at 0 °C for 20 minutes then warmed up to 50 °C and continued stirring under nitrogen atmosphere for 1.5 hours. The reaction mixture was cooled down to room temperature and 5M aqueous HCi was carefu!ly added until a clear solution was obtained. The reaction was stirred for 30 minutes. The reaction mixture was basified with 10M aqueous sodium hydroxide to reach pH 14 and the aqueous layer extracted with dich!oromethane:ethy! acetate (1 :1 , 2 x 100 mL). The combined organic layers were dried and evaporated to dryness. The residue was purified by chromatography on silica using a gradient elution from 0 to 100 % ethyl acetate in cyclohexane to afford the title compound (2.16 g, 6.35 mmol, 38 % yield) as a colourless oil. LC S (Method B): Rt 3.22 minutes; m/z 341 (MH+). intermediate 30:
-morpholinyimethyl)-4-pyrimidinarrtine
Figure imgf000098_0001
2-Chloro-6-(4-morpholiny!methyl)-/V-(1 ,1 ,3,3-tetramethy!butyi)-4-pyrimidinamine (2.16 g, 6.34 mmol) in trifluoroacetic acid (20 mL, 260 mmol) and dichloromethane (20 mL) was heated at 40 °C for 6 hours. The reaction mixture was then left standing at room temperature overnight. The reaction mixture was evaporated to dryness. The residue was partitioned between ethyl acetate:dichioromethane (1 : 1 , 100 mL) and saturated aqueous Na2C03 (30 mL). After separation, the aqueous phase was extracted with ethyl acetate (2 x 50 mL). The organic extracts were combined, dried using a hydrophobic frit and evaporated to dryness to afford the title compound (885 mg, 3.87 mmol, 61 % yield) as a white solid. LCMS (Method B): Rt 1.32 minutes; m/z 229 (MH+).
Intermediate 31 :
/V-[2-ch!oro-6-(4-morpholinylrnethyl)-4-pyrimidinyl][1,3]thiazolo[5,4-i)3pyridin-2-
Figure imgf000098_0002
Under an atmosphere of nitrogen, an ice-cooled solution of a mixture of 2-chloro-6-(4- morpholinylmethyl)-4-pyrimidinamine (1.14 g, 4.99 mmol) and 2-bromo[1 ,3]thiazolo[5,4- 0]pyridine (1.179 g, 5.48 mmo!) in dry Λ/,/V-dimethyiformamide (20 mL) was treated portionwise over 5 minutes with sodium hydride (60% w/w in mineral oil, 0.399 g, 9,97 mmo!). The reaction mixture was stirred with cooling for 1 hour and at ambient temperature for a further 1 hour. The mixture was treated cautiously with saturated ammonium chloride (5 mL). Saturated aqueous Na2C03 was added to reach pH 9 and the product was extracted with dichloromethane (2 x 100 mL). The organic extracts were combined, dried using a hydrophobic frit and evaporated to dryness. The residue was purified by chromatography on silica using a gradient elution from 0 to 100 % ethyl acetate in cyciohexane to afford the title compound (1.15 g, 3.17 mmol, 64 % yield). LCMS (Method B): Rt 1.82 minutes; m/z 363 (MH+).
Intermediate 32:
W2-{2-aminoethyl)-6-(4-morpho!inyim^
Figure imgf000099_0001
A microwave vial was charged with W-[2-chioro-6-(4-morpholinylmethyl)-4- pyrimidinyl][1 ,3]thiazolo[5,4-0]pyridin-2-am!ne (115 mg, 0.317 mmoi) and (2- aminoethy!)amine (0.023 mL, 0.349 mmol) in isopropanol (5 mL). The vial was sealed and was heated in a Biotage Initiator microwave at 160 °C for 30 minutes. The solvent was evaporated to dryness. The residue was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (31 mg, 0.08 mmol, 25 % yield). LCMS (Method B): Rt 1.67 minutes; m/z 387 (MH+).
in a second preparation, the dihydrochloride salt of the title compound was synthesised. Intermediate 33:
W2-[frans-2-aminocyclohexyI]-6-(4-morpholinyi
2-yl-2,4-pyrimidinediamine
Figure imgf000100_0001
A microwave vial was charged with W-[2-chloro-6-(4-morpholinylmethyi)-4- pyrimidinyl][1 ,3]thiazo!o[5,4-£]pyridin-2-amine (100 mg, 0.276 mmol) and trans-'] ,2- diaminocyciohexane (79 mg, 0.689 mmol) in isopropanol (4 mL). The vial was sealed and was heated in a Biotage Initiator microwave at 170 °C for 30 minutes. The solvent was evaporated to dryness. The residue was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (45 mg, 0.102 mmol, 37 % yield). LCMS (Method B): Rt 2.06 minutes; m/z 441 (MH+).
Intermediate 34:
1 ,1-dimethylethyl 3-{[4-(4-morpholinylmethyl)-6-{[1 ,3]thiazolo[5,4-fe]pyridin-2- ylamino)-2-pyrimidinyl]amino}-1-piperidinecarboxylate
Figure imgf000100_0002
A microwave vial was charged with W-[2-chloro-6-(4-morpholinylmethyl)-4- pyrimidiny!][1 ,3]thiazoio[5,4-/b]pyridin-2-amine (200 mg, 0.551 mmol) and 1 , 1 - dimethylethy! 3-amino~1 -piperidinecarboxyiate (221 mg, 1.102 mmol) in isopropanol (5 mL). The vial was sealed and was heated in a Biotage Initiator microwave at 170 °C for 40 minutes. A second portion of 1 ,1 -dimethylethyi 3-amino-1 -piperidinecarboxylate (110 mg, 0.55 mmol) was added and the reaction mixture heated in a Biotage Initiator microwave at 170 °C for 20 minutes. The solvent was evaporated to dryness. The residue was purified by chromatography on silica using a gradient e!ution from 0 to 15 % methanol (+1 % triethylamine) in dichloromethane to afford the title compound (225 mg, 0.427 mmol, 78 % yield). LCMS (Method B): Rt 2.54 minutes; m/z 527 (MH+). intermediate 35: 6-(4-morpholinylmethyl)-N2-3-piperi^
pyrimidinediamine dihydrochloride
Figure imgf000101_0001
A mixture of 1 , 1 -dimethylethyi 3-{[4-(4-morpho!inylrnethyl)-6-{t1 ,3]thiazolo[5,4-d]pyridin-2- ylamino)-2-pyrimidinyl]amino}-1 -piperidinecarboxylate (225 mg, 0.427 mmol) and 4M HCI in dioxane (1 mL, 4 mmol) in dichloromethane (5 mL) was stirred at room temperature for 16 hours. The solid was filtered under reduced pressure. The sticky solid was dissolved in methanof and the solvent evaporated to dryness to afford the title compound (200 mg, 0,4 mmolI 94 % yield). LCMS (Method B): Rt 1 .87 minutes; m/z 427 (MH+).
Intermediate 36:
W2-{ira 7s-4-aminocyclohexyl)-6-{4-morpholiny!methyl)-W4-[1 ,3]thiazolo[5
2-yl-2,4-pyrimidinediamine
Figure imgf000101_0002
A microwave vial was charged with A/-[2-chioro-6-(4-morphoiinyimethyl)-4- pyrimidinyi][1 ,3]thiazo!o[5,4-/b]pyridin-2-amine (200 mg, 0.551 mmol), irans-1 ,4- cyclohexanediamine (62.9 mg, 0.551 mmol) in isopropanol (4 mL). The vial was sealed and was heated in a Biotage Initiator microwave to 170 °C for 1 hour. After cooling, the reaction was filtered under reduced pressure and the solid was washed with isopropanol. The filtrate was evaporated to dryness to give yellow solid. The solid was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (55 mg, 0.125 mmol, 23 % yield). LCMS (Method B): Rt 1.81 minutes; m/z 441 (MH+). Intermediate 37:
1,1 -dimethylethyi (c s-4-{[4-(4-morpholinylmethyl)-6-([1,33thiazolo[5,4- )]pyridin-2- ylamino)-2-pyrimidinyl]amino}cyclohexy!)carbamate
Figure imgf000102_0001
A microwave vial was charged with A/-[2-chloro-6-(4-morpholinylmethyl)-4- pyrimidinyl][1 ,3]thiazolo[5,4- )]pyridin-2-arnine (200 mg, 0.551 mmol), 1 ,1 -dimethylethyl (c/s-4-aminocyclohexyl)carbamate (354 mg, 1.654 mmol) in isopropanol (4 mL). The vial was sealed and was heated in a Biotage Initiator microwave to 170 °C for 1 hour. The mixture was evaporated to dryness. The residue was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (135 mg, 0.25 mmol, 45 % yield). LCMS (Method D): Rt 1.07 minutes; m/z 541 ( H+).
Intermediate 38:
W2-(c;s-4-aminocyciohexyi)-6-(4-morphoH^^
yl-2,4-pyrimidtnediamine dihydrochloride
Figure imgf000102_0002
4M HCI in dioxane (0.375 mL, 1.498 mmol) was added to a solution of 1 ,1-dimethylethyl (c/s-4-{[4-(4-morphoiinylmethyl)-6-([1 ,3]thiazolo[5,4-0]pyridin-2-ylamino)-2- pyrimidinyl]amino}cyciohexyl)carbamate (135 mg, 0.25 mmol) in dichloromethane (3 mL). The reaction mixture was stirred at room temperature overnight. The reaction mixture was evaporated to dryness to afford the title compound (146 mg, 0.306 mmol, >100 % yield). LCMS (Method D): Rt 0.85 minutes; m/z 441 (MH+).
Intermediate 39:
1 ,1 -dimethylethyl 4-{t4-(4-morpholinylmethyl)-6-([1 ,3]thiazolo[5,4-Jb]pyridin-2- ylamino)-2-pyrimidinyl]amino}-1-piperidinecarboxylate
Figure imgf000103_0001
A microwave vial was charged with W-[2-chloro-6-(4-morpholinylmethyl)-4- pyrimidiny!][1 ,3]thiazolo[5,4-0]pyridin-2-amine (200 mg, 0.551 mmol) and 1 ,1- dimethylethyl 4-amino-1-piperidinecarboxylate (132 mg, 0.661 mmol) in isopropano! (5 mL). The via! was sealed and was heated in a Biotage Initiator microwave at 150 °C for 40 minutes. A further portion of 1 , 1-dimethylethyl 4-amino-1-piperidinecarboxylate (132 mg, 0.661 mmol) was added and the vial was heated at 170 °C for 30 minutes. The solvent was evaporated to dryness. The residue was purified by chromatography on silica using a gradient elution from 0 to 15 % methanol (+1 % triethy!amine) in dichioromethane to afford the title compound (235 mg, 0.447 mmol, 81 % yield). LCMS (Method B): Rt 2.64 minutes; m/z 527 (MH+). intermediate 40:
6-(4-morphoIinylmethyl)-W2-4-piperidm
pyrimidinediamine dihydrochloride
Figure imgf000103_0002
A mixture of 1 , 1-dimethylethyl 4-{[4-(4-morpholinylmethyi)-6-([1 ,3]thiazolo[5,4-0]pyridin-2- ylamino)-2-pyrimidinyl]amino}-1 -piperidinecarboxylate (235 mg, 0.446 mmol) and 4M HCi in dioxane (1 mL, 4 mmol) in dichioromethane (5 mL) was stirred at room temperature for 16 hours. The solid was filtered under reduced pressure and was dissolved in methanol. The solvent was evaporated to dryness to afford the title compound (240 mg, 0.481 mmol, >100 % yield). LCMS (Method B): Rt 1.76 minutes; m/z 427 (MH+).
Intermediate 41 :
1,1 -dimethylethyi (3S)-3-{[4-(4-morpholinylmethyl)-6-([1 )33thiazolo[5J4-b]pyridin-2- ylamino)-2-pyrimidinyl]amino}-1 »pyrrolidinecarboxy!ate
Figure imgf000104_0001
A microwave via! was charged with W-[2-chloro-6-(4-morpholinylmethy!)-4- pyrimidinylI[1 ,3]thiazolo[5,4-jb]pyndin-2-amine (200 mg, 0.551 mmol) and 1 ,1 - dimethylethy! (3S)-3-amino-1-pyrrolidinecarboxyiate (0.289 mL, 1.654 mmol) in isopropanol (4 mL). The vial was sealed and was heated in a Biotage Initiator microwave to 170 °C for 1 hour. The solution was evaporated to dryness. The residue was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (105 mg, 0.205 mmol, 37 % yield). LCMS (Method B): Rt 2.53 minutes; m/z 513 (MH+).
Intermediate 42:
6-(4-morpholinylmethyl)-W2-[{3S)-3-py^
e dihydrochloride
Figure imgf000104_0002
1 , 1 -Dimethylethyl (3S)-3-{[4-(4-morpholiny!methyi)-6-([1 ,3]thiazolo[5,4-£>]pyridin-2- ylamino)-2-pyrimidinyl]amino}-1-pyrro!idinecarboxylate (105 mg, 0.205 mmol) was dissolved in dichloromethane (3 mL) and methanol (1 mL) and treated with 4M HCI in dioxane (0.307 mL, 1.229 mmol). The reaction was stirred at room temperature overnight. The reaction mixture was evaporated to dryness to afford the title compound (106 mg, 0.218 mmol, >100 % yield). LCMS (Method D): Rt 0.77 minutes; m/z 413 (MH+).
Intermediate 43:
1,1 -dimethylethyl (3/?)-3-{[4-(4-morpholinylmethyl)-6-([1,3]thiazolo[5,4-it>]pyridm-2- ylamino)-2-pyrimidinyl]amino}-1 -pyrroiidinecarboxyiate
Figure imgf000105_0001
A microwave vial was charged with W-[2-chloro-6-(4-morpholinylmethyl)-4- pyrimidinylj[1 ,3jthiazolo[5,4-ib]pyridin-2-amine (200 mg, 0.551 mmol) and 1 ,1 - dimethylethyl (3R)-3-amino-1-pyrrolidinecarboxylate (0.289 mL, 1.654 mmol) in isopropanol (4 mL). The vial was sealed and was heated in a Biotage Initiator microwave to 170 °C for 1 hour. The solution was evaporated to dryness. The residue was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (97 mg, 0.189 mmol, 34 %). LCMS (Method B): Rt 2.53 minutes; m/z 5 3 (MH+).
Intermediate 44:
6-(4-morpholinylmethyl)-W2- ^^
2,4-pyrimidinediamine dihydrochloride
Figure imgf000105_0002
1 , 1 -Dimethylethyl (3 )-3-{[4-(4-morpholinylmethyj)-6-([1 ,3]thiazolo[5,4-/)3pyridin-2- ylamino)-2-pyrimidinyl]amino}-1-pyrrolidinecarboxylate (97 mg, 0.189 mmol) was dissolved in dichloromethane (3 mL) and methanol (1 mL) and treated with 4M HCI in dioxane (0.284 mL, 1.135 mmo!). The reaction was stirred at room temperature overnight. The reaction mixture was evaporated to dryness to afford the title compound (97 mg, 0.2 mmol, >100 % yield). LCMS (Method D): Rt 0.77 minutes; m/z 413 (MH+).
Intermediate 45:
W^c s-2-aminocyc!ohexyi]-6-(4-morpholiny^
yl-2,4-pyrimidinediamine
Figure imgf000106_0001
A microwave vial was charged with /V-[2-chloro-6-(4-morphoiinylmethyl)-4- pyrimidinyl][1 ,3]thiazolo[5,4-6]pyridin-2-amine (100 mg, 0.276 mmol) and c/s-1 ,2~diamino- cyclohexane (79 mg, 0.689 mmol) in isopropanol (4 mL). The vial was sealed and was heated in a Biotage Initiator microwave at 170 °C for 40 minutes. The solvent was evaporated to dryness. The residue was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (51 mg, 0.116 mmol, 42 % yield). LCMS (Method B): Rt 2.08 minutes; m/z 441 ( H+).
Intermediate 46:
W-{6-(3-amino-1-piperidinyl)-4-[(2-methyl-1 H-imidazol-1-yl)methyl]-^
bromo-1 ,3-benzothiazol-2-amine
Figure imgf000106_0002
A mixture of 6-bromo- \/-{6-chioro-4-[(2-methyl-1 H-imidazoS-1-yl)methyl]-2-pyrid!nyl}-1 l3- benzothiazol-2-amine (500 mg, 1 .15 mmol), 1 ,1-dimethylethyl 3-amino-1 - piperidinecarboxylate (1843 mg, 9.2 mmol) and ethylene glycol (1 mL) was heated at 195 °C under an atmosphere of nitrogen for 5 hours. Water (20 mL) was added and the solid was filtered under reduced pressure. The solid was washed with water, collected and dried under reduced pressure. The solid was triturated in ether, filtered under reduced pressure to afford the title compound (512 mg, 1 .027 mmol, 89 % yield) as a beige solid. LCMS (Method B): Rt 2.63 minutes; m/z 498, 500 (MH+). intermediate 47:
W-[6-(4-morphoiinylmethyl)-2-(1-piperazinyl}-4-pyrimidinyl3[1,3]thiazolo[5,4- b]pyridin-2-amine
Figure imgf000107_0001
A microwave vial was charged with /V-[2-chloro-6-(4-morpholinylmethyl)-4- pyrimidiny[j[1 ,3]thiazolo[5,4-b]pyridin-2-amine (115 mg, 0.317 mmol), piperazine (41.0 mg, 0.475 mmol) in isopropanol (4 ml_). The vial was sealed and was heated in a Biotage Initiator microwave to 150 °C for 30 minutes. The reaction mixture was evaporated to dryness. The residue was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (50 mg, 0.121 mmol, 38 % yield). LC S (Method D): Rt 0.73 minutes; m/z 413 (MH+). Intermediate 48:
1 ,1 -dimethylethyl 4-{[4-(4-morpholinylmethyl)-6-([1 ,3]thiazolo[5,4-i)]pyridin-2- ylamino)-2-pyrimidinyl]oxy}-1-piperidinecarboxylate
Figure imgf000107_0002
A microwave vial was charged with A/-[2-chloro-6-(4-morpholinylmethyl)-4- pyrimidinyl][1 ,3]thiazolo[5,4-jb]pyridin-2-amine (115 mg, 0.317 mmol), sodium hydride (60% w/w in mineral oil, 63.4 mg, 1 .585 mmol), 1 ,1-dimethylethyl 4-hydroxy-1- piperidinecarboxylate (191 mg, 0.951 tnmoS) and tetrahydrofuran (3 mL). The vial was sealed and was heated in a Biotage Initiator microwave to 130 °C for 45 minutes. The reaction mixture was partitioned between dichloromethane and saturated aqueous ammonium chloride. The organic layer was dried using a hydrophobic frit and evaporated to dryness. The residue was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (113 mg, 0.214 mmol, 68 % yield). LCMS (Method D): Rt 1.07 minutes; m/z 528 (MH+). Intermediate 49: N-[6-(4-morpholinyimethyl)-2-(4-piperi^
b]pyridin-2-amine dihydrochloride
Figure imgf000108_0001
1 , -Dimethylethyl 4-{[4-(4-morpholiny!methyl)-6-([1 ,3]thiazolof5,4-i^
pyrimidinyl]oxy}-1-piperidinecarboxylate (1 13 mg, 0.214 mmol) was dissolved in dichloromethane (3 ml_) and methanol (1 mL) and was treated with 4M HCI in dioxane (0.321 mL, 1.285 mmol). The reaction mixture was stirred at room temperature overnight. The reaction mixture was evaporated to dryness to afford the title compound (1 15 mg, 0.230 mmol, > 00 % yield). LCMS (Method B): Rt 1.68 minutes; m/z 428 <MH+).
Intermediate 50:
W2-(3-aminocyclohexyl)-6-(4-morphoiinylme^
-pyrimidinediamine
Figure imgf000108_0002
A microwave vial was charged with /V-[2-chioro-6-(4-morpholinylmethyl)-4- pyrimidinyl][1 ,3]thiazolo[5l4-i)]pyridin-2-amine (150 mg, 0.413 mmol), 1 ,3- cyc!ohexanediamine (1 18 mg, 1.034 mmol) and isopropanol (4 mL). The vial was sealed and was heated in a Biotage Initiator microwave at 170 °C for 30 minutes. The reaction mixture was evaporated to dryness. The residue was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (94 mg, 0.213 mmol, 52 % yield). LCMS (Method B): Rt 1.90 minutes; m/z 441 (MH+).
Intermediate 51 ;
W-[6-chloro-4-(4-morpholinylmethyl}-2-pyridinyl][1 ,3]thia2olo[5,4-J ]pyridin-2-amine
Figure imgf000109_0001
A mixture of 6-chioro-4-(4-morpholiny!methyl)-2-pyridinamine (0.96g, 4.22 mmol) and 2- bromo[1 ,3]thiazolo[5,4-j ]pyridine (1.088 g, 5.06 mmol) was dissolved in anhydrous N,N- dimethylformamide (10 mL), placed in an atmosphere of nitrogen and cooled in an ice bath. Sodium hydride (60% w/w in mineral oil, 0.337 g, 8.43 mmol) was added portionwise over 5 minutes and the mixture stirred for 1 hour whilst being allowed to warm to ambient temperature. The reaction mixture was then partitioned between dichloromethane and saturated aqueous ammonium chloride. The organic layer was dried using a hydrophobic frit, evaporated to dryness to give a yellow solid. The product was purified by chromatography on silica using a gradient elution from 0 to 100 % ethyl acetate in cyclohexane followed by 0 to 20 % methanol in ethyl acetate wash to afford the title compound (1 g, 2.763 mmol, 65 % yield) as a yellow solid. LC S: (Method B): Rt 2.51 minutes; m/z 362 ( H+). Intermediate 52:
W-[6-(3-amino-1 -piperidinyl)-4-(4-morphoIinylmethyl}-2-pyridinyl][1 ,3]thia
jb]pyridin-2-amine
Figure imgf000109_0002
A microwave vial was charged with 1 ,1 -dimethylethyl 3-amino-1 -piperidinecarboxylate (221 mg, 1.105 mmol), A/-[6-chloro-4-(4-morpholinyimethyl)-2-pyridinyl][1 ,3]thiazoio[5,4- /b]pyridin-2-amine (50 mg, 0.138 mmol) in ethylene glycol (1 ml). The vial was sealed and was heated in a Biotage Initiator microwave to 80 °C for 1 hour. A second portion of ,1- dimethylethyl 3-amino-1-piperidinecarboxyiate (221 mg, 1.105 mmol) was added and the vial was heated in a Biotage Initiator microwave at 220 °C for 1 hour. The residue was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (33 mg, 0.077 mmo!, 56 % yield). LCMS (Method B): Rt 2.06 minutes; m/z 426 (MH+). Intermediate 53:
N~[cis-3 -am i n ocy c lo h ex y 1] -4-{4-m orpholinylmethyl)-W-[1,33thiazolo[5,4- )]pyridin-2- yl-2,6-pyridinediamine
A microwave vial was charged with (3-aminocyclohexy!)amine (250 mg, 2.189 mmol), Λ/- [6-chloro-4-(4-morpholinylmethyl)-2-pyridinyl][1 ,3]thiazolo[5,4-jb]pyridin-2-amine (59 mg, 0.163 mmol) in ethylene glycol (1 ml). The vial was sea!ed and was heated in a Biotage Initiator microwave to 200 °C for 1 hour. A second portion of (3-aminocyclohexyl)amine (250 mg, 2.189 mmoi) was added to the reaction mixture and the microwave vial was heated in a Biotage Initiator microwave system for 1 hour at 220 °C. A third portion of (3- aminocyclohexyi)amine (250 mg, 2.189 mmoi) was added to the reaction mixture and the microwave vial was heated in a Biotage Initiator microwave for 1 hour at 220 °C. The residue was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (14 mg, 0.032 mmoi, 20 % yield). LC S (Method B): Rt 2.02 minutes; m/z 440 (MH+). intermediate 54:
W-[frarts-3-aminocyclohexyl]-4-(4-morpholinylm
2-yi-2,6-pyridinediamine
Figure imgf000110_0002
A microwave vial was charged with (3-aminocyciohexyl)amine (250 mg, 2.189 mmol), N- [6-ch!oro-4-(4-morpho]inylmethyl)-2-pyridinyl][1 ,3]thiazolo[5,4-b]pyridin-2-amine (59 mg, 0.163 mmol) in ethylene giycol (1 ml). The vial was sealed and was heated in a Biotage Initiator microwave to 200 °C for 1 hour. A second portion of (3-aminocyclohexyl)amine (250 mg, 2.189 mmol) was added to the reaction mixture and the microwave vial was heated in a Biotage initiator microwave system for 1 hour at 220 °C. A third portion of (3- aminocyclohexy!)amine (250 mg, 2.189 mmol) was added to the reaction mixture and the microwave vial was heated in a Biotage Initiator microwave for 1 hour at 220 °C. The residue was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (10 mg, 0.023 mmoi, 14 % yield). LCMS (Method B): Rt 2.06 minutes; m/z 440 (MH+).
Intermediate 55:
W-[c s-2-aminocyclohexyl]-4-(4-morpho^
yl-2,6-pyridinediamine
Figure imgf000111_0001
A microwave vial was charged with W-[6-ch!oro-4-(4-morpholinylmethyl)-2- pyridinylj[1 ,3Jthiazoio[5,4-J ]pyridin-2-amine (100 mg, 0.276 mmol), c/s-1 ,2- cyclohexanediamine (410 mg, 3.59 mmol) in ethylene glycol (1 ml). The vial was sealed and was heated in a Biotage Initiator microwave to 220 °C for 1 hour. The residue was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (18 mg, 0.041 mmol, 15 % yield). LCMS (Method B): Rt 2.20 minutes; m/z 440 (MH+).
Intermediate 56:
^[frans-2-aminocyc!ohexyI]-4-(4-morpholinylm
2-yl-2,6-pyridinediamine
Figure imgf000111_0002
A microwave vial was charged with A/-[6-chloro-4-(4-morpholinyimethyl)-2- pyridinyl][1 ,3]thiazolo[5,4-/b]pyridin-2-amine (100 mg, 0.276 mmol), ira/?s-1 ,2- cyclohexanediamine (31.6 mg, 0.276 mmol) in ethylene glycol (1 ml). The vial was sealed and was heated in a Biotage initiator microwave to 220 °C for 1 hour. The residue was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (20 mg, 0.045 mmol, 16 % yieid). LCMS (Method B): Rt 2.17 minutes; m/z 440 (MH+).
Intermediate 57:
1 ,1-dimethylethyl methyl(2-{[4-(4-morpholinylmethyl)-6-([1,3]thiazolo[5,4-b]pyridin- 2-yiamino)-2-pyrimidinyl]amino}ethyl)carbamate
Figure imgf000112_0001
A microwave vial was charged with A/-[2-chioro-6-(4-morpholinylmethyl)-4- pyrimidinyl][1 ,3]thiazolo[5,4-0]pyridin-2-amine (150 mg, 0.413 mmol) and 1 ,1 - dimethylethyl (2-aminoethyl)methylcarbamate (79 mg, 0,455 mmol) in isopropanol (5 mL). The vial was sealed and was heated in a Biotage Initiator microwave at 160 °C for 30 minutes. A second portion of 1 , 1-dimethylethyl (2-aminoethyl)methylcarbamate (79 mg, 0,455 mmol) was added and the vial was heated in a Biotage Initiator microwave at 150 °C for 1 hour. The residue was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (57 mg, 0.114 mmol, 28 % yield). LCMS (Method B): Rt 2,58 minutes; m/z 501 (MH+). Intermediate 58:
Wz-[2-(methylamino)ethyl]-6-(4-morpholm^
yl-2,4-pyrimidinediamine dihydrochloride
Figure imgf000112_0002
4 HCI in dioxane (0.150 mL, 0.599 mmol) was added to a solution of 1 , -dimethyiethyl methyl(2-{[4-(4-morpholinylmethyl)-6-([1 [3]thiazolo[5,4-b]pyridin-2-ylamino)-2- pyrimidinyi]amino}ethyl)carbamate (50 rng, 0.100 mmoi) in dichioromethane (3 mL) and methanol (1 mL). The reaction mixture was stirred at room temperature overnight. The reaction mixture was evaporated to dryness to afford the title compound (50 mg, 0.106 mmol, >100 % yield). LCMS (Method B): Rt 1.88 minutes; m/z 401 (MH+).
Intermediate 59:
1 ,1-dimethylethyl {3-[4-(4-morpholinylmethyl)-6-([1 ,3]thia2oio[5,4-i)]pyridin-2- ylamino)-2-pyrimidinylJphenyl}carbamate
Figure imgf000113_0001
A microwave vial was charged with A/-[2-chloro-6-(4-morpholinyimethyl)-4- pyrimidinyl][1 ,3]thiazo!o[5,4-i)]pyndin-2-amine (200 mg, 0.551 mmol), [3-({[(1 , 1- dimethylethyl)oxy]carbonyl}amino)pheny[jboronic acid (261 mg, 1.102 mmol), chioro[2'- (dimethylamino)-2-biphenylyl]pa!ladium - (1 R,4S)-bicydo[2.2.1 ]hept-2-yl[(1 S,4f?)- bicyclo[2.2.1]hept-2-yl]phosphane (1 : 1 ) (30.9 mg, 0.055 mmol) and tripotassium phosphate (468 mg, 2.205 mmo!) in 1 ,4-dioxane (4 ml) and water (1 ml). The vial was sealed and was heated in a Biotage Initiator microwave system at 160 °C for 1.5 hours. After cooling, the reaction mixture was partitioned between dichioromethane and saturated aqueous sodium bicarbonate. The organic extract was dried using a hydrophobic frit and evaporated to dryness. The residue was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (20 mg, 0.038 mmol, 7 % yield). LCMS (Method B): Rt 2.99 minutes; m/z 520 (MH+).
Intermediate 60:
6-bromo-W-{6-chloro-4-[(2-methy 1-1 W-imidazol-1 -yl)methyl]-2-pyridinyl}-1 ,3- benzothiazol-2-amine
Figure imgf000114_0001
Under an atmosphere of nitrogen, an ice-coo!ed, stirred suspension of 6-chloro-4-[(2- methyl-1 H-imidazol-1-yi)methyl]-2-pyridinamine (1.6 g, 7.19 mmol) in anhydrous teirahydrofuran (40 mL) was treated with 6-bromo-2-chloro-1 ,3-benzothiazoie (1.96 g, 7.90 mmol) and, portionwise over 5 minutes, with sodium hydride (60 % w/w in mineral oil, 0.632 g, 15.8 mmol). After 30 minutes, the mixture was heated to 50 °C for 6 hours. The cooled mixture was treated with saturated aqueous ammonium chloride (30 mL) and tetrahydrofuran (30 mL). The resulting precipitate was filtered off, washed with ethyi acetate and diethyl ether and dried to afford the title compound (2.2 g, 5.06 mmol, 70 % yield). LCMS (Method A): Rt 0.94 minutes; m/z 434, 436 (MH+).
Intermediate 61 :
-methyl-1 H-imidazol-1-y!)rnethyl]-2-pyridinamine
Figure imgf000114_0002
A mixture of 2,6-dichloro-4-[(2-methyl-1 H-imidazol-1-yl)methy!]pyridine (3.6 g, 14.9 mmol) and concentrated aqueous ammonia (10 mL, 517 mmol) was sealed and heated in a Biotage Initiator microwave at 160 °C (pressure limited to 19 bar) for 9 hours. The cooled suspension was added to water (120 mL) and then filtered. The filtered solid was washed with water and dried to afford the title compound (3.1 g, 13.9 mmol, 94 % yield). LCMS (Method A): Rt 0.33 minutes; m/z 223, 225 (MH+).
Intermediate 62:
-dichloro-4-[{2-methyl-1 H-imidazol-1 -yl)methyl]pyridine
Figure imgf000114_0003
Under an atmosphere of nitrogen, an ice-cooled solution of 2-methyl-1 H-imidazole (1.7 g, 20.7 mmol) in tetrahydrofuran (30 mL) was treated portionwise with sodium hydride (60% in mineral oil, 1 g, 25.0 mmol). After 15 minutes, the mixture was treated with 2,6-dichloro- 4-(chloromethyl)pyridine (4 g, 20.4 mmol) and then allowed to warm slowly to ambient temperature whereupon it was stirred for 8 hours. The mixture was then treated with saturated aqueous ammonium chloride (40 mL) and extracted with ethyl acetate (2 x 40 mL). The combined organics were dried over magnesium sulfate, filtered and evaporated to dryness. The product was purified by chromatography on silica using a gradient elution from 0 to 100 % ethyl acetate in cyclohexane followed by a gradient of 0 to 20 % methanol in dichloromethane to afford the title compound (3.7 g, 15.3 mmol, 75 % yield). LCMS (Method A): Rt 0.47 minutes; m/z 242, 244 (MH+).
Intermediate 63:
morpholinylmethyl)-2-pyridinamine
Figure imgf000115_0001
A solution of 4-[(2,6-dichloro-4-pyridinyl)methyl]morpholine (2 g, 8.1 mmol) in ethylene glycol (5 mL) was treated with concentrated aqueous ammonia (5.7 mL, 262 mmol) and the mixture was sealed and heated in a Biotage Initiator microwave at 200 °C for 24 hours. The cooled reaction mixture was filtered and the solid was dried, then suspended in diethyl ether, filtered and dried to afford the title compound (816 mg, 3.6 mmol, 44 % yield). LCMS (Method A): Rt 0.29 minutes; m/z 228 (MH+).
Intermediate 64:
4-[(2,6-dichloro-4-pyridinyl)methyl]morpholine
Figure imgf000115_0002
Under an atmosphere of nitrogen, an ice-coo!ed solution of 4-[(2,6-dichloro-4- pyridinyl)carbonyl]morpho!ine (2.63 g, 10.1 mmol) in dichloromethane (50 mL) was treated with a solution of borane in tetrahydrofuran (1 M, 30.2 mL, 30.2 mmol) and stirred with cooling for 1 hour. Additional borane in tetrahydrofuran (30.2 mL, 30.2 mmol) was added and the reaction was stirred until complete. The mixture was then carefully acidified with 5M aqueous hydrochloric acid and then heated at reflux. The reaction mixture was then neutralised with 10 % aqueous sodium hydroxide. The aqueous phase was extracted twice with ethyl acetate (+10 % methanol) and the combined organic fraction was dried over magnesium sulfate, filtered and evaporated to dryness to afford the title compound (2.89 g, 11.7 mmol, >100 % yield). LCMS (Method A): Rt 0.49 minutes; m/z 248, 250 (MH+).
Intermediate 65:
-[(2,6-dichloro-4-pyridinyI)carbonyl]morpholine
Figure imgf000116_0001
Morpholine (2.36 g, 27.1 mmol) was added to a stirred ice-cooled mixture of 2,6-dichloro- 4-pyridinecarbonyl chloride (2.85 g, 13.5 mmol) and pyridine (5.70 ml_, 70.4 mmol) in dichloromethane (5 ml_) and the mixture was stirred with cooling for 2 hours. The mixture was treated with saturated aqueous sodium hydrogen carbonate (20 mL) and then extracted with ethyl acetate (2 x 20 mL). The combined organic fractions were washed with brine (20 mL), dried over magnesium sulfate, filtered and the solvent evaporated to dryness. The residue was then purified by chromatography on silica using a gradient elution from 0 to 100 % ethyl acetate in cyclohexane to afford the title compound (2.93 g, 11.2 mmol, 83 % yield). LCMS (Method A): Rt 0.76 minutes; m/z 261 ,263 (MH+). Intermediate 66:
4-{4-morpholinylmethyl)-2,6-pyridinediamine
Figure imgf000116_0002
A microwave vial was charged with 4-[(2,6-dichioro-4-pyridinyl)methyl]morpho!ine (2.5 g, 10.12 mmol), copper powder (200 mg, 3.15 mmol) and aqueous ammonia (10 mL, 517 mmol). The vial was sealed and heated in a Biotage Initiator microwave at 170 °C (maximum pressure 18 bars) for 15 hours. Copper powder (200 mg, 3.15 mmol) was added and the sealed vial was heated in a Biotage Initiator microwave at 170 °C (maximum pressure 18 bars) for further 15 hours. The reaction mixture was filtered on celite and the fiitrate was evaporated to dryness. The product was purified by reverse phase (C18) chromatography using a gradient elution from 0 to 50 % acetonitri!e in water with an ammonium bicarbonate modifier to afford the title compound (1.02 g, 4.9 mmol, 48 % yield). LCMS (Method D): Rt 0.47 minutes; m/z 209 (MH+).
Intermediate 67:
-(4-morpholinylmethyl)-A/-[1 ,3]i/) azo/o[5,4-fa]pyridin-2-yl-2I6-pyridinedi
Figure imgf000117_0001
A round bottom flask was charged with 4-(4-morpholinylmethy!)-2,6-pyridinediamine (900 mg, 4.32 mmol) and 2-bromo[1 ,3]thiazolo[5,4-jb]pyridine (1.022 g, 4.75 mmol) in anhydrous N,/V-dimethy!formamide (10 mL). The reaction was placed in an atmosphere of nitrogen and cooled in an ice bath. Sodium hydride (60 % w/w in mineral oil, 519 mg, 12.96 mmol) was added portionwise over 5 minutes and the mixture stirred for 1 hour whilst being allowed to warm to ambient temperature. The reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was then partitioned between dichloromethane and water. After separation, the organic !ayer was dried using an hydrophobic frit and evaporated to dryness to give a black oil. The product was purified by chromatography on aminopropyl (NH2) silica using a gradient elution from 0 to 100 % ethyl acetate In cyc!ohexane to afford the title compound (417 mg, 1.218 mmol, 28 % yield) as an off-white soiid. LCMS: (Method D): Rt 0.74 minutes; m/z 343 (MH+).
Intermediate 68:
1,1 -dimethy lethyl (3S)-3-{{4-[(6-bromo-1 ,3-benzothiazol-2-yl)amino]-2- pyrimidinyl}amino)-1-pyrrolidinecarboxylate
Figure imgf000117_0002
A microwave vial was charged with 6-bromo-A/-(2-chloro-4-pyrimidinyl)-1 ,3-benzothiazol- 2-amine (500 mg, 1.464 mmol), 1 ,1-dimethylethyl (3S)-3-amino-1-pyrrolidinecarboxylate (0.383 mL, 2.195 mmol) and isopropanol (5 mL). The microwave vial was sealed and heated in a Biotage Initiator microwave at 170 °C for 45 minutes. The solvent was evaporated to dryness to give the title compound (720 mg, 1.465 mmol, 100 % yield) as a white solid. LCMS: (Method D): Rt 1.26 minutes; m/z 91 , 493 (MH+). intermediate 69:
6-bromo-W-[2-chloro-6-{4-morpholinylmeth^
amine
Figure imgf000118_0001
In an atmosphere of nitrogen, an ice-cooled solution of a mixture of 2-ch!oro-6-(4- morpholinylmethyl)-4-pyrimidinamine (1.356 g, 5.93 mmol) and 6-bromo-2-chloro-1 ,3- benzothiazo!e (1 ,474 g, 5.93 mmol) in dry A/,A/-dimethy!forrnamide (30 mL) was treated portionwise over 5 minutes with sodium hydride (60% w/w in mineral oil, 0.474 g, 1 1.86 mmol). The reaction mixture was stirred with cooling for 1 hour and at ambient temperature for a further 1 hour. The mixture was treated cautiously with saturated ammonium chloride (7.5 mL). Saturated aqueous sodium carbonate was added (aqueous pH=11 ) and the product was extracted with dichloromethane (2 x 100 mL). The organic layers were combined, dried using a hydrophobic frit and evaporated to dryness. The residue was purified by chromatography on silica using a gradient elution from 0 to 100 % ethyl acetate in cyciohexane followed by 0 to 30 % methanol in dichloromethane to afford the title compound (1.7 g, 3.86 mmol, 65 % yield). LCMS (Method D): Rt 1.19 minutes; m/z 440, 442 (MH+).
Intermediate 70:
1 ,1 -dimethylethyl (2-{[4-[(6-bromo-1 ,3-benzothiazol-2-yl)amino]-6-{4- morpholinylmethyl)-2-pyrimidinyl]amino}ethyl)carbamate
Figure imgf000118_0002
A microwave vial was charged with 6-bromo-A/-[2-chloro-6-(4-morpholinylmethyl)-4- pyrimidinyl]-1 ,3-benzothiazol-2-amine (500 mg, 1.134 mmol), 1 , 1 -dimethylethyl (2- aminoethyl)carbamate (0.539 mL, 3.40 mmol) and isopropanol (5 mL). The microwave vial was sealed and heated in a Biotage Initiator microwave at 170 °C for 45 minutes. The solid was filtered under reduced pressure to give the titie compound (384 mg, 0.680 mmol, 60 % yield) as an off-white solid. LCMS (Method D): Rt 1 .19 minutes; m/z 564, 566 (MH+).
Intermediate 71 :
1,1 -dimethylethyl (3S)-3-{[4-[(6-bromo-1 ,3-benzothiazoI-2-yl)amino]-6-(4- -pyrimidinyl]amino}-1 -pyrrolidinecarboxylate
Figure imgf000119_0001
A microwave vial was charged with 6-bromo-/V-[2-chloro-6-(4-morpholinylmethyl)-4- pyrimidinyl]-1 ,3-benzothiazol-2-amine (500 mg, 1.134 mmol), 1 ,1 -dimethylethyl (3S)-3- amino- -pyrrolidinecarboxylate (0.297 mL, 1.702 mmol) and isopropanol (5 mL). The microwave via! was sealed and heated in a Biotage Initiator microwave at 170 °C for 45 minutes. 1 ,1 -Dimethylethyl (3S)-3-amino-1-pyrrolidinecarboxylate (0.297 mL, 1.702 mmol) was added, the vial was sealed and heated in a Biotage Initiator microwave at 170 °C for further 45 minutes. The solvent was evaporated to dryness. The residue was purified by chromatography on silica using a gradient elution from 0 to 25 % methanol in dichloromethane to afford the title compound (402 mg, 0.68 mmol, 60 % yield) as a brown solid. LCMS (Method D): Rt 1.28 minutes; m/z 590, 592 (MH+).
Intermediate 72:
1,1 -dimethylethyl 4-{[4-(4-morpholiny!methyl)-6-([1 ,3]thiazolo[5,4-i)]pyridin-2- ylamino)-2-pyridinyi]amino}-1-piperidinecarboxyiate
Figure imgf000120_0001
A microwave vial was charged with A/-[6-chloro-4-(4-morpholinylmethyl)-2- pyridiny!]E1 ,3]thiazolo[5,4-Jb]pyridin-2-amine (50 mg, 0.138 mmol), 1 ,1 -dimethylethyi 4- amino-1 -piperidinecarboxylate (41.5 mg, 0.207 mmol), {1 ,3-bis[2,6-bis(1 - methylethyl)phenyl]-2-imidazolidinyl}(chloro)(2-methyl-2-propen-1-yS)palladium (24.40 mg, 0.041 mmol). The system was sealed and placed under an atmosphere of nitrogen using a vacuum purge. Lithium bis(trimethylsiiyi)amide, 1 M in tetrahydrofuran (0.5 mL, 0.5 mmol) was added. The vial was stirred in the preheated oil bath at 80 °C for 1 hour. The reaction was cooled down to room temperature. The reaction mixture was partitioned between dichloromethane (10 mL) and saturated aqueous ammonium chloride (10 mL). After separation, the organic extract was dried using a hydrophobic frit and evaporated to dryness. The residue was purified by chromatography on silica using a gradient elution from 0 to 15 % methanol (+1 % triethyiamine) in dichloromethane to afford the title compound (52 mg, 0.099 mmol, 72 % yield) as a light brown solid. LCMS (Method D): Rt 1.1 1 minutes; m/z 526 (MH+).
Intermediate 73:
1 ,1 -dimethylethyl {3S)-3-{i4-(4-morpholinylmethyl)-6-([1 ,3]thiazolo[5,4-Jb]pyndin-2- ylamino)-2-pyridinyl]amino}-1-pyrrolidinecarboxylate
Figure imgf000120_0002
A microwave vial was charged with /V-[6-chloro-4-(4-morpholinylmethyl)-2- pyridinyl][1 ,3]thiazolo[5,4-b]pyridin-2-amine (100 mg, 0.276 mmol), 1 ,1 -dimethylethyl (3S)- 3-amino-1-pyrrolidinecarboxylate (77 mg, 0.415 mmol), {1 ,3-bts[2,6-bis(1 - methylethyl)phenyl]-2-imidazolidinyl}(chloro)(2-methyl-2-propen-1-yi)palladium (48.8 mg, 0.083 mmol). The system was sealed and placed under an atmosphere of nitrogen using a vacuum purge. Lithium bis(trimethy!silyl)amide, 1 M in tetrahydrofuran (1 mL, 1 mmol) was added. The vial was stirred in the preheated oil bath at 80 °C for 1 hour. The reaction was cooled down to room temperature. The reaction mixture was partitioned between dichloromethane (10 mL) and saturated aqueous ammonium chloride (10 mL). After separation, the organic extract was dried using a hydrophobic frit and evaporated to dryness. The residue was purified by chromatography on silica using a gradient elution from 0 to 15 % methanol (+1 % triethylamine) in dichloromethane to afford the title compound (133 mg, 0.26 mmol, 94 % yield) as a light brown solid. LCMS (Method D): Rt 1.06 minutes; m/z 512 (MH+).
Intermediate 74:
4-(4-morphoiinyimethyl)~W-3-piperidm
pyridinediamine
Figure imgf000121_0001
A microwave vial was charged with A/-[6-ch!oro-4-(4-morpholinylmethyl)-2- pyridinyl][1 ,3]thiazolo[5,4-i»]pyridin-2-amine (100 mg, 0.276 mmol), 1 ,1 -dimethyiethyl 3- amino-1-piperidinecarboxylate (83 mg, 0.415 mmol), {1 ,3-bis[2,6-bis(1- methyiethyl)phenyl]-2-imidazolidinyl}(chioro)(2-methyi-2-propen-1-yl)palladium (48.8 mg, 0.083 mmol). The system was sealed and placed under an atmosphere of nitrogen using a vacuum purge. Lithium bis(trimethy!silyl)amide, 1 M in tetrahydrofuran (1 mL, 1 mmoi) was added. The vial was stirred in the preheated oil bath at 80 °C for 4 hours. The reaction was cooled down to room temperature. The reaction mixture was partitioned between dichloromethane (10 mL) and saturated aqueous ammonium chloride (10 mL). The acqeous phase was basified to pH 13 with saturated aqueous sodium bicarbonate and was extracted with dichloromethane:methanol (9: 1 , 3 x 20 mL). The organic extracts were combined, dried using a hydrophobic frit and evaporated to dryness to afford the title compound (58 mg, 0.136 mmo!, 49 % yield) as a yeliow solid. LCMS (Method D): Rt 0.77 minutes; m/z 426 (MH+).
Intermediate 75: 2-chioro-6-{[c s-2,6-dimethyl-4-morpholinyl]ca^
Figure imgf000122_0001
A round bottom flask was charged with methy! 2-chloro-6-[(1 , 1 ,3,3- tetramethylbutyl)amino]-4-pyrimidinecarboxylate (18.95 g, 63.2 mmol) and tetrahydrofuran (150 mL). The solution was stirred at 0 °C under nitrogen atmosphere during 10 minutes. C/s-2,6-dimethylmorphoiine (7.83 mL, 63.2 mmol) and 1 ,3,4,6, 7,8-hexahydro-2H- pyrimido[1 ,2-a]pyrimidine (2.64 g, 18.96 mmoi) were added. The reaction mixture was stirred for 15 hours under nitrogen atmosphere. The mixture was partitioned between dichloromethane (70 mL) and water (70 mL). After separation, the organic layer was dried using a hydrophobic frit and evaporated to dryness to afford the title compound (23.31 g, 60.9 mmol, 96 % yield) as ao off-white solid. LCMS (Method D): Rt 1.33 minutes; m/z 383 (MH+). Intermediate 76:
2-chloro-6-{[c/s-2,6-dimethyl-4-morphoM
pyrimidinamine
Figure imgf000122_0002
1 M Borane in tetrahydrofuran (183 mL, 183 mmol) was added dropwise to a solution of 2- chloro-6-{[c/s-2,6-dimethyl-4-morpholinyl]carbony!}-A/-(1 , 1 ,3,3-tetramethylbutyl)-4- pyrimidinamine (23.31 g, 60.9 mmol) stirring at 0 °C in tetrahydrofuran (40 mL) under nitrogen atmosphere. The reaction was stirred at 0 °C for 20 minutes then warmed up to 50 °C and continue stirring under nitrogen atmosphere for 1 ,5 hours. The reaction mixture was cooied down to 0 °C. 5M aqueous HCI was carefully added until a clear solution was obtained. The reaction mixture was stirred for 1 hour and then was basified with 10M aqueous sodium hydroxide to reach pH=12. The product was extracted with dichloromethane (2 x 100 mL). 5M aqueous HCI (50 mL) was carefully added to the organic extracts and the reaction mixture was stirred for 1 hour. The reaction mixture was basified with 10M aqueous sodium hydroxide to reach pH 12. After separation, the aqueous phase was back extracted with dichioromethane (2 x 100 mL). The combined organic layers were dried using a hydrophobic frit and evaporated to dryness. The residue was purified by chromatography on silica using a gradient elution from 0 to 100 % ethyl acetate in cyclohexane to afford the title compound (8.25 g, 22.4 mmol, 37 % yield) as a colourless oil. LCMS (Method D): Rt 1.41 minutes; m/z 369 (MH+). intermediate 77:
-chloro-6-{[c/s-2,6-dimethyl-4-morphoiinyl]methyi}-4-pyrimidinam
Figure imgf000123_0001
2-Chloro-6-{[c/s-2>6-dimethyl-4-morpho!inyl]methyl}-/\/-(1 l1 ,3,3-tetramethy!buty!)-4- pyrimidinamine (8.25 g, 22,36 mmol) in trifluoroacetic acid (200 mL, 2596 mmol) and dichioromethane (200 mL) was heated at 40 °C for 6 hours and the reaction mixture was left standing at room temperature overnight. The solvent was evaporated to dryness. The residue was partitioned between ethyl acetate (200 mL) and saturated aqueous sodium carbonate (200 mL). After separation, the aqueous phase was back extracted with ethyl acetate (100 mL). The organic extracts were combined, dried using a hydrophobic frit and evaporated to dryness. The residue was purified by chromatography on silica using a gradient elution from 0 to 10 % methanol in dichioromethane to afford the title compound (3.5 g, 13.6 mmol, 61 % yield) as a white solid. LCMS (Method D): Rt 0.66 minutes; m/z 257 (MH+).
Intermediate 78:
W-(2-chloro-6-{[c/*s-2f6-dimethyl-4-morpholinyI]methyl}-4- pyrimidinyl)[1 ,3]thiazoio[5,4-f>]pyridin-2-amine
Figure imgf000123_0002
Under an atmosphere of nitrogen, an ice-cooled solution of 2-chloro-6-{[c/s-2,6-dimethyi- 4-morpholinyl]methyl}-4-pyrimidinamine (500 mg, 1.948 mmol) and 2- bromo[1 ,3]thiazoio[5,4-6]pyridine (419 mg, 1.948 mmol) in dry V,A/-dimethyjformamide (10 mL) was treated portionwise over 5 minutes with sodium hydride (60 % w/w in mineral oil) (156 mg, 3.90 mmol). The reaction mixture was stirred with cooiing for 30 minutes and at ambient temperature for a further 1 hour. The mixture was treated cautiously with saturated aqueous ammonium chloride (3 ml_). Saturated aqueous sodium carbonate was added (aqueous pH 1 1 ) and the product was extracted with dichloromethane (2 x 50 m!_).The solvent was evaporated to dryness to afford the title compound (767 mg, 1.96 mmol, >100 % yield) as a white solid. LCMS (Method D): Rt 0.87 minutes; m/z 391 (MH+).
Intermediate 79:
1 ,1-dimethylethyl 4-{[4-{[c/s-2,6-dimethyl-4-rnorpholinyl3methyl}-6-([1 ,3]thiazolo[5,4- ft]pyridin-2-ylamino)-2-pyrimidinyl3amino}-1-piperidinecarboxylate
Figure imgf000124_0001
A microwave via! was charged with A/-(2-chloro-6-{[c/'s-2,6-dimethyl-4-morpholinyl]methyl}- 4-pyrimidinyl)[1 ,3jthiazolo[5,4-i)]pyridin-2-amine (150 mg, 0.384 mmol), 1 ,1-dimethylethyl 4-amino-1-piperidinecarboxylate (231 mg, 1.151 mmol) and isopropanol (5 ml_). The vial was sealed and heated in a Biotage Initiator microwave at 170 °C for 45 minutes. 1 ,1- dimethylethyl 4-amino-1 -piperidinecarboxylate (116 mg, 0.579 mmol) was added. The vial was sealed and heated in a Biotage Initiator microwave at 170 °C for 45 minutes. The solvent was evaporated to dryness. The residue was purified by chromatography on silica using a gradient eiution from 0 to 25 % methanol in dichloromethane to afford the title compound (148 mg, 0.267 mmol, 70 % yield) as a brown solid. LCMS (Method D): Rt 1.16 minutes; m/z 555 (MH+).
Intermediate 80:
1,1 -dimethylethyl (3S)-3-{[4-{[c/'s-2,6-dimethyl-4-morpholinyl]methyl}-6- ([1 ,3]thiazolo[5,4-i)]pyridin-2-yiamino)-2-pyrimidinyl]amino}-1 - pyrroiidinecarboxylate
Figure imgf000125_0001
A microwave vial was charged with W-(2-chloro-6-{[c/s-2,6-dimethyl-4-morpholinyl]rnethyl}- 4-pyrimidinyl)[1 ,3]thtazolo[5>4-jb]pyridin-2-amine (150 mg, 0.384 mmol), 1 ,1-dimethylethyl (3S)-3-amino-1 -pyrrolidinecarboxyiate (0.201 mL, 1.151 mmol) and isopropanoi (5 mL). The microwave vial was sealed and heated in a Biotage Initiator microwave at 170 °C for 45 minutes. The solvent was evaporated to dryness. The residue was purified by chromatography on siiica using a gradient elution from 0 to 25 % methanol in dich!oromethane to afford the title compound (1 17 mg, 0.216 mmol, 56 % yield) as a brown gum. LCMS (Method D): Rt 1.10 minutes; m/z 541 (MH+). intermediate 81 :
1 ,1-dimethylethyl (2-{[4-{[c s-2s6-dimethyl-4-morpholinyl3methyl}-6- din-2-yiamino}-2-pyrimidinyl]amino}ethyl)carbamate
Figure imgf000125_0002
A microwave vial was charged with A/-(2-chloro-6-{[c/5-2)6-dimethyl-4-morpholinyl]methyl}- 4-pyrimidinyl)[1 I3]thiazoio[5,4-i)]pyridin-2-amine (150 mg, 0.384 mmol), 1 ,1 -dimethylethyl (2-aminoethyl)carbamate (0.182 mL, 1 , 151 mmol) and isopropanoi (5 mL). The microwave vial was sealed and heated in a Biotage Initiator microwave at 170 °C for 45 minutes. The solvent was evaporated to dryness. The residue was purified by chromatography on siiica using a gradient elution from 0 to 25 % methanol in dichloromethane to afford the title compound (134 mg, 0.26 mmol, 68 % yield) as an off- white solid. LCMS (Method D): Rt 1.0 minutes; m/z 515 (MH+). Intermediate 82:
o-4^yridinyI)carbonyf]-c s-2,6-dimethylmorpholine
Figure imgf000126_0001
C/'s-2,6-dimethy!morpholine (7.06 mL, 57 mmol) was added dropwise to a stirred, cooled 0 °C mixture of 2,6-dichloro-4-pyridinecarbonyl chloride (10 g, 47.5 mmol) and N,N- diisopropylethylamine (24.9 mL, 143 mmol) in dichloromethane (30 mL). The reaction mixture was stirred at 0 °C for 2 hours and then at room temperature for 16 hours. Aqueous saturated ammonium chloride (70 mL) was added to the reaction and the product was extracted with dichloromethane (100 mL). The organic extract was washed with aqueous saturated ammonium chloride (70 mL), was dried using a hydrophobic frit and evaporated to dryness to afford the title product (14.4 g, 49.8 mmol, >100 % yield) as a yellow solid. LCMS (Method D): Rt 0.97 minutes; m/z 289 (MH+). Intermediate 83:
o-4-pyridinyl)methyl]-c s-2,6-dimethyimorpholine
Figure imgf000126_0002
1 Borane in tertrahydrofuran (149 mL, 149 mmol) was added dropwise to a cooled solution (0 °C) of 4-[(2,6-dichloro-4-pyridinyl)carbonyl]-c s-2,6-dimethylmorpholine (14.4 g, 49.8 mmol) in dichloromethane (50 mL). The reaction was stirred at 0 °C for 1 hour under nitrogen atmosphere. 5 aqueous HC! was carefully added to the reaction while still stirring at 0 °C (carefull effervescence). After stirring for 2 hours, the solution was left standing at room temperature overnight. The mixture was basified with 10M aqueous sodium hydroxide to reach pH 14. The product was extracted with dichloromethane (50 mL). The organic phase was dried using a hydrophobic frit and evaporated to dryness. The residue was purified by chromatography on silica using a gradient elution from 0 to 100 % ethyl acetate in dichloromethane to afford the title compound (9.9 g, 36 mmol, 72 % yield) as an off-white solid. LCMS (Method D): Rt 1.19 minutes; m/z 275 / 277 (MH+). Intermediate 84:
-2,6-dimethyl-4-morpholinyl3methyl}-2-pyridinam
Figure imgf000127_0001
A solution of 4-[(2,6-dichloro-4-pyndinyl)methyl]-c s-2,6-dirnethylmorpholine (3.5 g, 12.72 mmoi) in isopropanol (3 ml_) was treated with concentrated aqueous ammonia (7 mL, 323 mmo!). The reaction mixture was heated at 160 °C in a Biotage Initiator microwave for 30 hours (maximum pressure 18 bars). The reaction mixture was evaporated to dryness. The residue was purified by chromatography on silica using a gradient elution from 0 to 100 % ethyl acetate in cyclohexane followed by 0 to 20 % methanol in ethyl acetate to afford the title compound (960 mg, 3.75 mmol, 29 % yield), LCMS (Method D): Rt 0.85 minutes; m/z 254 (MH-).
Intermediate 85:
W-{6-chloro-4-((c/'s-2,6-dimethylmorpholino)methyl)pyridin-2-yl)thiazolo[5,4- i)]pyridin-2-amine
Figure imgf000127_0002
To an ice cooled solution of 6-chloro-4-((c/s-2,6-dimethylmorpholino)methyl)pyridin-2- amine (960 mg, 3,75 mmol) and 2-bromo[1 ,3]thiazoio[5,4-/t>]pyridine (888 mg, 4.13 mmoi) in N, Ν-ά i m ethy if orm amide (10 mL) was added sodium hydride (60 % w/w in mineral oil, 300 mg, 7.51 mmol) portionwise. The reaction was stirred at 0 °C under nitrogen atmosphere for 1 hour and at room temperature for another 1 hour. Saturated aqueous ammonium chloride was added and the product was extracted with ethyl actetate. After separation, the organic extract was dried using a hydrophobic frit and evaporated to dryness. The residue was purified by chromatography on silica using a gradient elution from 0 to 25 % methanol in dichloromethane to afford the title compound (1.38 g, 3.54 mmol, 94 % yield) as a brown solid. LCMS (Method D): Rt 1 , 17 minutes; m/z 390 (MH+). intermediate 86: 1 ,1-dimethylethyl 4-{[4-{[c/s-2,6-dimethyi-4-morpholinyl]methyl}-6-([1 ,3]thiazolo[5,4- dinyl]amino}-1 -piperidinecarboxylate
Figure imgf000128_0001
A microwave vial was charged with A/-(6-chloro-4-((c/s-2,6- dimeihyimorpholino)methyl)pyridin-2-y[)thiazolo[5,4-J ]pyridin-2-amine (150 mg, 0.385 mmoi), 1 ,1 -dimethyiethyi 4-amino-1 -piperidinecarboxylate (116 mg, 0.577 mmol) and {1 ,3- bis[2,6-bis(1-methyiethyi)phenyl]-2-imidazo!idinyl}(chloro)(2-methyl-2-propen-1- yi)paliadium (67.9 mg, 0.1 15 mmol). The vial was sealed and purged with nitrogen and vacuum. Lithium bis(trimethylsily!)amide, 1 M in tetrahydrofuran (1 .5 mL, 1.5 mmol) was added. The vial was stirred in the preheated oil bath at 80 °C for 1 hour. The reaction mixture was cooled down to room temperature. The reaction mixture was partitioned between dichloromethane (10 mL) and saturated aqueous ammonium chloride (10 mL). After separation, the organic extract was dried using a hydrophobic frit and evaporated to dryness. The residue was purified by chromatography on silica using a gradient elution from 0 to 25 % methanol in dichloromethane to afford the title compound (213 mg, 0.385 mmol, 100 % yield) as a brown solid. LC S (Method D): Rt 1.22 minutes; m/z 554 (MH+).
Intermediate 87:
1 ,1 -dimethylethyl (3S)-3-{[4-{[c s-2,6-dimethyl-4-rnorpholinyI]methyt}-6- idin-2-ylamino)-2-pyridm^
Figure imgf000128_0002
A microwave vial was charged with W-(6-chioro-4-((c s-2,6- dimethylmorpholino)methyl)pyridin-2-yl)thiazolo[5,4-£>]pyridin-2-amine (150 mg, 0.385 mmol), 1 , 1-dimethylethyl (3S)-3-amino-1 -pyrrolidinecarboxylate (107 mg, 0.577 mmo!) and {1 ,3-bis[2,6-bis(1-methylethyl)phenyl3-2-imidazo!idinyl}(chloro)(2-meth
yl)palladium (68 mg, 0.1 15 mmol). The system was sealed and placed under an atmosphere of nitrogen using a vacuum purge. Lithium bis(trimethy!silyl)amide, 1 M in tetrahydrofuran (1.5 mL, 1 ,5 mmol) was added. The vial was stirred in the preheated oil bath at 80 °C for 1 hour. The reaction was cooled down to room temperature. The reaction mixture was partitioned between dichloromethane (10 mL) and saturated aqueous ammonium chloride (10 mL). After separation, the organic extract was dried using a hydrophobic frit and evaporated to dryness. The residue was purified by chromatography on silica using a gradient elution from 0 to 25 % methanol in dichloromethane to afford the title compound (208 mg, 0.385 mmol, 100 % yield) as a brown solid. LCMS (Method D): Rt 1.16 minutes; m/z 540 (MH+).
Intermediate 88:
1,1 -dimethyiethyl 3-{[4-{[c/s-2)6-dίmethyl·4-morpholinyi]methyl}-6-([1 ,3]thiazolo[5,4- b]pyridin-2-ylamino)-2-pyridinyl]amino}-1-piperidinecarboxylate
Figure imgf000129_0001
A microwave vial was charged with W-(6-chloro-4-((c/s-2,6- dimethylmorpholino)methyl)pyridin-2-yl)thtazolo[5,4-03pyridin-2-amine (150 mg, 0.385 mmol), 1 ,1 -dimethylethyl 3-amino-1 -piperidinecarboxylate (1 16 mg, 0.577 mmol) and {1 ,3- bis[2,6-bis(1 -methylethyl)pheny!]-2-imidazolidinyl}(ch!oro)(2-methyi-2-propen-1- yl)pa!ladium (67.9 mg, 0.1 15 mmol). The system was sealed and placed under an atmosphere of nitrogen using a vacuum purge. Lithium bis(trimethy!silyl)amide, 1 M in tetrahydrofuran (1.5 mL, 1.5 mmol) was added. The vial was stirred in the preheated oil bath at 80 °C for 1 hour. The reaction mixture was cooled down to room temperature and was partitioned between dichloromethane (10 mL) and saturated aqueous ammonium chloride (10 mL). After separation, the organic extract was dried using a hydrophobtc frit and evaporated to dryness. The residue was purified by chromatography on silica using a gradient elution from 0 to 25 % methanol in dichloromethane to afford the title compound (100 mg, 0.181 mmol, 47 % yield) as a brown solid. LCMS (Method D): Rt 1.18 minutes; m/z 554 ( H+).
Intermediate 89:
1 ,1 -dimethylethyl 3-{[4-(4-morpholinylmethyl)-6-([1,3]thiazoIo[5,4-ib]pyridin-2- ylamino)-2-pyridinyl]amino}-1 -piperidinecarboxylate
Figure imgf000130_0001
A microwave vial was charged with A/-[6-ch!oro-4-(4-morpholinylmethyl)-2- pyridinyl][1 ,3]thiazolo[5,4-jt>]pyridin-2-amine (100 mg, 0.276 mmol), 1 ,1 -dimethylethyl 3- amino-1 -piperidinecarboxylate (83 mg, 0.415 mmol) and {1 ,3-bis[2,6-bis(1- methylethyl)phenyl]-2-imidazolidinyi}(ch!oro)(2-methyl-2-propen-1 -yl)pa!ladium (49 mg, 0.083 mmo!). The system was sealed and placed under an atmosphere of nitrogen using a vacuum purge. Lithium bis(trimethylsilyl)amide, 1 in tetrahydrofuran (1 mL, 1 mmol) was added. The via! was stirred in the preheated oil bath at 70 °C for 1 hour. The vial was cooled down to room temperature. The reaction mixture was partitioned between dichloromethane (10 mL) and saturated aqueous ammonium chioride (10 mL). After separation, the organic extract was dried using a hydrophobic frit and evaporated to dryness. The residue was purified by chromatography on silica using a gradient elution from 0 to 15 % methanol (+ 1 % triethy!amine) in dichloromethane to afford the title compound (80 mg, 0.152 mmol, 55 % yield). LCMS (Method D): Rt 1.07 minutes; m/z 526 (MH+).
Intermediate 90:
2-amino-6-chloro-4-pyrtdinecarboxyiic acid
Figure imgf000130_0002
A mixture of 2,6-dichloro-4-pyridine carboxylic acid (110 g, 0.58 mol) and aqueous ammonia solution (26 %, 440mL) was sealed and heated in a microwave reactor (split into 11 separate vessels) at 165 °C (pressure limited to 19 bars) for 8 hours. The cooled mixtures were combined and evaporated to dryness. The residue was dissolved in water (2.5 L) and the pH was adjusted to 3 by the addition of concentrated aqueous hydrochloric acid. The resulting precipitate was filtered, washed with water and thoroughly dried to afford the title compound (100 g, 0.58 mol, 100 % yield). LCMS (Method A): Rt 0.52 minutes; m/z 173, 175 (MH+). intermediate 91 :
(2-amino-6-chloro-4-pyridinyl)methanol
Figure imgf000131_0001
Under an atmosphere of nitrogen and at ambient temperature, a solution of lithium aluminium hydride (98.2 g, 1.25 mol) in tetrahydrofuran (2200 mL) was treated portionwise with 2-amino-6-chloro-4-pyridine carboxylic acid (98.2 g, 569 mmol) whilst maintaining the temperature of the reaction below 20 °C. The mixture was stirred overnight at ambient temperature and then cooled to 0 °C. Water (70 mL) was slowly added dropwise. After a further 5 minutes, aqueous potassium hydroxide solution (15 % w/v, 70 mL) was slowly added, followed by additional water (200 mL). The mixture was allowed to warm to ambient temperature before magnesium sulfate was added and then stirred for 30 minutes. The mixture was then filtered, the filtered solid was washed with ethyl acetate (2000 mL) and the filtrate was evaporated to dryness to afford the crude product. The crude product was suspended in dichloromethane (500 mL), filtered and the filtered solid was dried to afford the title compound (80 g, 0.506 mol, 88 % yield). LCMS (Method A): Rt 0.66 minutes; m/z 159 (MH+).
Intermediate 92:
6-chloro-4- 1 ,1-dimethylethyi){dimethyl)silyl]oxy}methyl)-2-pyridinamine
Figure imgf000131_0002
Under an atmosphere of nitrogen and at ambient temperature a stirred solution of (2- amino-6-chloro-4-pyridinyl)methanol (48 g, 0.30 mol) and imidazole (24.6 g, 0.36 mol) in anhydrous A/,A/-dimethylformamide (800 mL), was treated with chioro(1 ,1 ~ dimethyiethyi)dimethylsilane (46.4 g, 0.31 moS). The reaction mixture was stirred for 90 minutes and then concentrated in vacuo whilst maintaining the temperature below 40 °C. Water was added and the mixture was extracted with dichloromethane (3 x 500 mL). The combined organics were washed with water, dried over magnesium sulfate and filtered through a pad of silica. The filtrate was evaporated to dryness to afford the title compound (49.3 g, 0.18 mol, 60% yield). LCMS (Method A): Rt 1.58 minutes; m/z 273 (MH+).
Intermediate 93:
W-te-chloro^-U i.l-dimethylethylKdimethylJsilylloxylmethyl)^- pyridin-2-amine
Figure imgf000132_0001
Under an atmosphere of nitrogen, an ice-cooied mixture of 6-chloro-4-({[(1 , 1- dimethylethy!)(dimethyl)silyl]oxy}methyj)-2-pyridinamine (2 g, 7.33 mmol) and 2- bromo[1 ,3]thiazoio[5,4-fo]pyridine (1 .58 g, 7.33 mmol) in anhydrous N,N- dimethyiformamide (20 mL) was treated portionwise over 5 minutes with sodium hydride (60% w/w in mineral oil, 0.586 g, 14.7 mmol) and the mixture was stirred for 3 hours whilst being allowed to warm to ambient temperature. The mixture was then treated cautiously with saturated aqueous ammonium chloride (20 mL) and water (30 mL). The precipitated solid was filtered, washed with water and dried to afford the title compound (2.87 g, 7.05 mmol, 96% yield). LCMS (Method A): Rt 1.51 minutes; m/z 407 (MH+).
Intermediate 94:
1 ,1 -dimethylethyl 4-{[4-({[(1,1 -dimethylethyl)(dimethyl)si!yl]oxy}methyl)-6- 1 ,3]thiazolo[5,4-b]pyridin-2-ylamino}-2-pyridinyI]amino}-1-piperidinecarboxylate
Figure imgf000132_0002
A microwave vial was charged with /V-[6-chloro-4-({[(1 ,1- dimethylethy[)(dimethyl)silyl]oxy}methyl)-2-pyridinyl][1 ,3]thiazolo[5,4-i)]pyridin-2-amine (1 g, 2.457 mmol), 1 ,1 -dimethylethyl 4-amino-1-piperidinecarboxylate (0.738 g, 3.69 mmol) and {1 ,3-bis[2,6-bis(1 -methylethyl)phenyi]-2-imidazoiidinyl}(chloro)(2-methyl-2-propen-1 - y!)palladium (0.434 g, 0.737 mmo!). The system was sealed and placed under an atmosphere of nitrogen using a vacuum purge. Lithium bis(trimethylsiiy!)amide, 1 M in tetrahydrofuran (10 mL, 10 mmol) was added. The vial was stirred in the preheated oil bath at 70 °C for 1 hour. The reaction was cooled down to room temperature. The reaction mixture was partitioned between dichloromethane (50 mL) and saturated aqueous ammonium chloride (50 mL). After separation, the organic extract was dried using a hydrophobic frit and evaporated to dryness. The residue was purified by chromatography on silica using a gradient elution from 0 to 100 % ethyl acetate in cyclohexane to afford the title compound (846 mg, 1.48 mmol, 60 % yield) as a yellow solid. LCMS (Method D): Rt 1.57 minutes; m/z 571 (MH+).
Intermediate 95:
1,1 -dimethylethyl 4-{[4-(hydroxymethyl)-6-([1,3]thia2olo[5!4-b]pyridin-2-ylamino)-2- pyridinyI]amino}-1-piperidinecarboxylate
Figure imgf000133_0001
To a solution of 1 , 1-dimethylethyl 4-{[4-({[(1 , 1-dimethyiethyl)(dimethyl)silyl]oxy}methyl)-6- ([1 ,3]thiazolo[5,4-i)]pyridin-2-ylamino)-2-pyridinyl]amino}-1-pipehdinecarboxy!ate (840 mg, 1.472 mmol) in tetrahydrofuran (15 mL) stirred at room temperature was added tetrabutylammonium fluoride, 1 M in tetrahydrofuran (2.207 mL, 2.207 mmol). The reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was partitioned between dichloromethane (50 mL) and water (50 mL). The organic phase was dried using a hydrophobic frit and evaporated to dryness to afford the title compound (680 mg, 1.415 mmol, 96 % yield). LCMS (Method D): Rt .04 minutes; m/z 457 (MH+).
Intermediate 96:
1 ,1-dimethylethyl 4-{t4-formyl-6-(t1,3]thiazolo[5,4-b3pyridin-2-ylamino)-2- pyridinyljaminoH -piperidinecarboxyiate
Figure imgf000134_0001
To a solution of 1 ,1 ,1-triacetoxy-1 , 1-dihydro-1 ,2-benziodoxol-3(1 H)-one (67 mg, 0.153 mmol) in tetrahydrofuran (1.5 mL), stirred at 0 °C, was added a solution of 1 ,1 - dimethylethy! 4-{[4-(hydroxymethyl)-6-([1 ,3]thiazolo[5,4-i>]pyridin-2-ylamino)-2- pyridinyl]amino}-1 -piperidinecarboxylate (50 mg, 0.11 mmol) in tetrahydrofuran (1.5 mL). The reaction mixture was warmed to room temperature and stirred for 30 minutes. The reaction mixture was partitioned between dichioromethane (10 mL) and water (10 mL). After separation, the aqueous phase was washed with dichioromethane (10 mL). The organic phases were combined, dried using a hydrophobic frit and evaporated to dryness. The residue was purified by chromatography on silica using a gradient elution from 0 to 100 % ethyi acetate in cyclohexane followed by 0 to 20 % methanol in ethyl acetate to afford the title compound (48 mg, 0.084 mmol, 77 % yield) as a yellow solid. LCMS (Method D): Rt 1.15 minutes; m/z 455 (MH+). Intermediate 97:
1,1 -dimethyIethyi (3S)-3-{[4-{{[(1 ,1 -dimethylethyl)(dimethyl)silyl]oxy}methyl)-6- ( 1 ,3]thtazolo[5,4-/i]pyridin-2-ylamino)-2-pyridi
Figure imgf000134_0002
A microwave via! was charged with A/-[6-chloro-4-({[(1 , 1- dimethylethyl)(dimethyi)silyl]oxy}methyl)-2-pyridinyl][1 ,3]thiazolo[5,4-jb]pyridin~2-amine
(3.25 g, 7.99 mmol), 1 ,1 -dimethylethyl (3S)-3-amino-1-pyrrolidinecarboxylate (2.231 g, 1 1.98 mmol) and {1 ,3-bis[2,6-bis(1 -methylethy phenyl]-2-imidazolidinyl}(chloro)(2-methy!- 2-propen-1-yl)palladium (1.410 g, 2.396 mmol). The system was sealed and piaced under an atmosphere of nitrogen using a vacuum purge. Lithium bis(trimethylsi!yi)amide, 1 M in tetrahydrofuran (60 mi, 60 mmol) was added. The vial was stirred in the preheated oil bath at 70 °C for 1 hour. The reaction was cooled down to room temperature. The reaction mixture was partitioned between dichloromethane (150 mL) and saturated aqueous ammonium chloride (150 mL). After separation, the organic extract was dried using a hydrophobic frit and evaporated to dryness. The residue was purified by chromatography on silica using a gradient elution from 0 to 100 % ethyl acetate in cyclohexane to afford the title compound (2.04 g, 3.66 mmol, 46 % yield) as a red-brown solid. LCMS (Method D): Rt 1.52 minutes; m/z 557 (MH+).
Intermediate 98:
1,1 -dimethylethyl (3S)-3-{[4-(hydroxymethyl)-6-([1,3]thiazolo[5,4-i)]pyridin-2- yiamino)-2-pyridiny!]amino}-1 -pyrrolidinecarboxylate
Figure imgf000135_0001
To a stirred solution of 1 , 1-dimethylethyl (3S)-3-{[4-({[(1 , 1- dimethylethyl)(dimethyl)silyl]oxy}methyl)-6-([1 ,3]thiazolo[5,4-jb]pyhdin-2-ylamino)-2- pyridinyl]amino}-1-pyrrolidinecarboxylate (2.04 g, 3.66 mmol) in tetrahydrofuran (20 mL) at room temperature was added tetrabutylammonium fluoride, 1 M in tetrahydrofuran (5.5 mL, 5.5 mmol). The reaction mixture was stirred at room temperature for 1 hour. The reaction was cooled down in an ice bath and water (130 mL) was added. The suspension was stirred in the ice bath for 10 minutes and was filtered under reduced pressure. The solid was washed with water and dried in the oven to afford the title compound (1.43 g, 3.23 mmol, 88 % yield) as a brown solid. LCMS (Method D); Rt 0.98 minutes; m/z 443 (MH+).
Intermediate 99:
1 ,1 -dimethyiethyl (3S)-3-{[4-formy l-6-([1 ,3]thiazolo[5,4-b|pyridin-2-y lamino)- pyridinyl]amino}-1-pyrrolidinecarboxylate
Figure imgf000136_0001
To a solution of 1 , 1 -triacetoxy-1 , 1-dihydro-1 ,2-benziodoxol-3(1 H)-one (1 ,978 g, 4.52 mmol) in tetrahydrofuran (30 mL), stirred at 0 °C, was added a solution of 1 , 1- dimethyiethyl (3S)-3-{[4-(hydroxymethyl)-6-([1 ,3]thiazolo[5,4-i ]pyridin-2-ylamino)-2- pyridiny!]amino}-1-pyrroiidinecarboxylate (1.43 g, 3.23 mmol) in tetrahydrofuran (30 mL). The reaction mixture was warmed to room temperature and stirred for 0.5 hour. The reaction mixture was partitioned between dichloromethane (100 mL) and water (100 mL). The organic phase was dried using a hydrophobic frit and evaporated to dryness. The residue was purified by chromatography on silica using a gradient elution from 0 to 100 % ethyl acetate in cyc!ohexane to afford the title compound (610 mg, 1.385 mmol, 43 % yield) as a brown solid. LCMS (Method D): Rt 1.09 minutes; m/z 41 (MH+).
Intermediate 100:
4-{[c/s-2,6-dimethy!-4-morpholinyl]m
-b3pyridin-2-yl-2,6-pyridinediamine dihydrochloride
Figure imgf000136_0002
1 , 1 -Dimethylethyl (3S)-3-{[4-{[c/s-2,6-dimethyl-4-morphoiinyl3methyi}-6-([1 ,3]thiazolo [5,4- d]pyridin-2-ylamino)-2-pyrimidinyl]amino}-1-pyrroiidinecarboxylate (36.2 g, 67.1 mmol) was dissolved in dichloromethane (250 mL) and methanol (10 mL). The stirred reaction mixture was cooled to 0 °C and 4 HCI in dioxane (120 mL, 480 mmol) was added dropwise. The reaction mixture was then stirred at room temperature for 16 hours. The yellow solid was filtered under reduced pressure, washed with dichloromethane (2 x 100 mL) and dried in an oven at 40 °C for 16 hours to afford the crude title compound (37 g, 72.2 mmol, > 100 % yield) as a yellow solid which was used directly without purification. LCMS (Method E): Rt 0.52 minutes; m/z 440 (MH+). Intermediate 101 :
4-{[c/s-2,6-dimethyl-4-morpholinyl]^
-b]pyridin-2-yl-2,6-pyridinediamine
Figure imgf000137_0001
4-{[c s-2,6-Dimethyl-4-morpholiny!]methyl^
b]pyridin-2-yl-2,6-pyridinediamine dihydrochloride (2 g, 3,90 mmol) was dissolved in dichloromethane (20 mL) and methanol (20 mL). MP-carbonate (6.48 g, 19.51 mmol) was added and the reaction was stirred at room temperature for 3 hours. The MP-carbonate resin was filtered under reduced pressure, washed with dichloromethane/methanol (1 :1 , 20 mL) and the filtrate was evaporated to dryness to afford the title compound (1.67 g, 3.80 mmol, 97 % yield) as an off-white solid. LC S (Method E): Rt 0.53 minutes; m/z 440 (MH+)
Intermediate 102:
1 ,1 -dimethylethyl 3-{[4-(4-morpholinylmethyl)-6-([1,3]thiazolo[5,4-b]pyridin ylamino)-2-pyridinyl]amino}-1-azetidinecarboxylate
Figure imgf000137_0002
/V-[6-Chloro-4-(4-morpholinylmethyl)-2-pyhdinyl][1 ,3]thiazolo[5,4-/b]pyridin-2-amine (100 mg, 0.276 mmol) and 1 ,1-dimethylethy! 3-amino-l-azetidinecarboxylate (71 .4 mg, 0.415 mmol), were dissolved in tetrahydrofuran (1 mL) in a microwave vial. The system was sealed and placed under an atmosphere of nitrogen using a vacuum purge (3 times). {1 ,3- Bis[2[6-bis(1 -methylethyl)phenyl]-2-imidazoiidinyl}(chloro)(2-methyl-2-propen-1 - yl)palladium (48.8 mg, 0.083 mmol) was dissolved In a mixture of tetrahydrofuran (1 mL) and lithium bis(trimethylsiiyl)amide, 1 M in THF (0.829 mL, 0.829 mmol) which was then added to the microwave vial. The reaction mixture was then stirred at 50 °C for 4 hours. On cooling, the reaction mixture was partitioned between dichloromethane (30 mL) and saturated aqueous ammonium chloride (30 mL). After separation, the organic extract was dried using a hydrophobic frit and evaporated to dryness. The residue was purified by chromatography on silica using a gradient elution from 50 to 100 % ethyl acetate in dichloromethane. The appropriate fractions were combined and evaporated in vacuo to give a light brown solid. The brown solid was triturated in acetonitrile (10 mL). The solid was then filtered under reduced pressure, washing with acetonitrile to afford the title compound (53 mg, 0.106 mmol, 38 % yield) as a brown solid. LCMS (Method D): Rt 1.06 minutes; m/z 498 (MH+). intermediate 103:
2 6-dichloro-W,W-dimethyl-4-pyridinecarboxamide
Figure imgf000138_0001
To a solution of 2,6-dichloroisonicotinic acid (10 g, 52.1 mmol) in Λ/,/V-dimethylformamide (50 mL) stirred at room temp was added 1 , 1 '-carbonyldiimidazole (10.13 g, 62.5 mmol) in a single portion. The reaction mixture was stirred under nitrogen at room temperature for 1 hour while dimethylamine hydrochloride (5.48 mL, 62.5 mmol) and triethylamine (14.52 mL, 104 mmol) were dissolved and stirred in A/,W-dimethylformamide (30 mL) at room temperature. Then, the two solutions were combined and the resultant reaction mixture was stirred under nitrogen at room temperature for 2 hours. A further portion of 1 ,1 '- carbonyldiimidazole (2.53g, 15.62mmol) was added, and the reaction mixture was stirred for 1 hour at room temperature. The reaction mixture was partitioned between dichloromethane (500 mL) and saturated aqueous sodium carbonate (500 mL). After separation, the organic extract was washed with saturated aqueous sodium carbonate (200 mL) and aqueous ammonium chloride (200 mL) then dried using a hydrophobic frit and evaporated to dryness. The residue was purified by chromatography on silica using a gradient elution from 0 to 100 % ethyi acetate in cyclohexane to afford the title compound (10.05 g, 45.9 mmol, 88 %) as a white solid. LCMS (Method D): Rt 0.80 minutes; m/z 219/221 (MH+).
Intermediate 104:
1 -(2,6-dichloro-4-pyridinyl)-W,W-dimethylmethanamine
Figure imgf000139_0001
Borane-tetrahydrofuran complex, 1 M in tetrahydrofuran (150 mL, 150 mmol) was added dropwise to a cooled solution (0 °C) of 2,6-dichloro-A/,W-dimethyl-4-pyridinecarboxamide (14.6 g, 66.6 mmol) in dichloromethane (150 mL) under a nitrogen atmosphere over 1 hour. After the addition, the reaction was stirred at 0 °C for a further 1 hour then at room temperature for 64 hours. More borane-tetrahydrofuran complex, 1 M in tetrahydrofuran (66.6 mL, 66.6 mmol) was then added and the reaction was left stirring for 4 hours. 2 M aqueous HCI (170 mL, 340 mmol) was carefully added to the reaction while stirring at 0 °C (effervescence). After stirring for 1 hour while maintaining the temperature between 0 - 5 °C, the solution was left stirring at room temperature for 1 hour. The mixture was basifted with 10 M aqueous sodium hydroxide (60.0 mL, 600 mmol) to reach pH 12, then left stirring for 1 hour. The mixture was separated and the organic phase was dried using a hydrophobic frit and concentrated under reduced pressure to give the crude product. The residue was purified by chromatography on silica using a gradient elution from 0 to 100 % ethyl acetate in cyciohexane. Further purification was conducted via reverse phase chromatography using a C18 column using a gradient elution from 0 to 100% acetonitrile (+ 0.1 % ammonia) in water (+ 0.1 % ammonia) to give the title compound (3.92 g, 19.1 mmol, 29 %). LCMS (Method D): Rt 1.03 minutes; m/z 205 (MH+). Intermediate 105:
-chloro-4-[(dimethylamino)methyl]-2-pyridinamine
Figure imgf000139_0002
A pressure vessel was charged with 1-(2,6-dichloro-4-pyridinyl)-A/,W-dimethy! methanamine (3.92 g, 19.1 1 mmol), isopropanoi (10 mL) and 35% aqueous ammonia solution (8.34 mL, 431 mmol). The vessel was sealed and heated at 170 °C for 18 hours (pressure 20 bar). After cooling to room temperature, the residue was partitioned between ethyl acetate and 1 M aqueous sodium hydroxide, After separation, the organic layer was dried using a hydrophobic frit and evaporated to dryness. The residue was purified by chromatography on silica using a gradient elution from 0 to 15 % methanol in dichloromethane. Further purification was conducted via chromatography on silica using a gradient elution from 0 to 100% ethy! acetate in dichloromethane followed by 0 to 20 % methanol in dichloromethane gradient to give the title compound (1 g, 5.39 mmo!, 28 %) as a brown solid. LCMS (Method D): Rt 0.73 minutes; m/z 186 (MH+).
Intermediate 106:
W-{6-chIoro-4-[(dimethylamino)methyl]-2^
mine
Figure imgf000140_0001
A round bottom flask was charged with sodium ferf-butoxide (494 mg, 5.14 mmol) in dry tetrahydrofuran (3 mL), placed under an atmosphere of nitrogen and cooled in an ice bath. 2-Bromo[1 ,3]thiazolo[5,4-/b]pyridsne (371 mg, 1.725 mmol) and 6-chloro-4- [(dimethylamino)methyl]-2-pyridinamine (308 mg, 1.659 mmoi) dissolved in dry tetrahydrofuran (4 mL) were then added dropwise and the mixture stirred for 1 hour at 0- 10 °C, then 20 hours at room temperature. A further 0.5 equivalents of sodium tert- butoxide (80 mg, 0.83 mmol) was added and the reaction mixture was stirred for 3 more hours at room temperature. Saturated aqueous ammonium chloride (10 ml) was added while the reaction was stirred at room temperature. The mixture was then extracted with dichloromethane (30 mL). The two phases were separated and the organic phase was dried using a hydrophobic frit and evaporated to dryness to give the crude product as a brown solid. The residue was triturated with acetonitrile (10 mL). The resulting solid was filtered through a Buchner funnel, washed with acetonitrile (5 mL), and collected to give the title compound (385 mg, 1.2 mmol, 73 % yield) as an off-white solid. LCMS (Method D): Rt 1.05 minutes; m/z 320 (MH+).
Intermediate 107:
1,1 -dimethylethyl (3S)-3-{[4-[(dimethylamino)methyl]-6-([1 ,3]thiazolo[5,4-Jb] pyridine- 2- lamino)-2-pyridinyl]amino}-1-pyrrolidinecarboxylate
Figure imgf000140_0002
A microwave vial was charged with W-{6-chioro-4-[(dimethyiamino)methyl]-2- pyridinyl}[1 ,3]thiazoloI5,4-6]pyriclin-2-amine (150 mg, 0.469 mmol), 1 ,1 -dimethylethyl (3S)- 3-amino-1-pyrrolidinecarboxyiate (131 mg, 0.704 mmoi) and dissolved in teirahydrofuran (1 mL). The system was sealed and placed under an atmosphere of nitrogen using a vacuum purge (3 times). {1 ,3-Bis[2,6-bis(1 -methylethyl)phenyl]-2-imidazolidinyl} (ch!oro)(2-methyl-2-propen-1-yl)palladium (83 mg, 0.141 mmoi) was mixed with lithium bis(trimethylsiiy!)amide, M in tetrahydrofuran (1.407 mL, 1.407 mmol), then added to the reaction mixture. The reaction mixture was stirred at 50 °C for 2 hours. The reaction was cooled to room temperature then partitioned between dichioromethane (30 mL) and saturated aqueous ammonium chloride (30 mL). After separation, the organic extract was dried using a hydrophobic frit and evaporated to dryness. The residue was purified by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (81 mg, 0.172 mmol, 37%). LC S (Method D): Rt 1.10 minutes; m/z 470 (MH+). intermediate 108:
1 ,1-dimethylethyl (3f?)-3-{[6-([1 ,3]thiazolo[5,4-i ]pyridin-2-ylamino)-4-({[(1 S)-1 ,2,2- trimethyIpropyl]amino}methyl)-2-pyridinyl]amino}-1 -pyrrolidinecarboxylate
Figure imgf000141_0001
A microwave vial was charged with /V-[6-chioro-4-({[(1 S)-1 )2,2- trimethylpropyl]amino}methyl)-2-pyridinyl][1 ,33thiazoio[5,4-/b]pyridin-2-amine (100 mg, 0.266 mmol), 1 , 1-dimethylethyl (3R)-3-amino-1-pyrroiidinecarboxylate (74.3 mg, 0.399 mmoi), dissolved in tetrahydrofuran (1 mL). The system was sealed and placed under an atmosphere of nitrogen using a vacuum purge (3 times). {1 ,3-Bis[2,6-bis(1 - methylethyl)phenyl]-2-imidazolidinyl}(chloro)(2-methyl-2-propen-1-yl)palladium (47 mg, 0.08 mmol) was mixed with lithium bis(trimethylsilyl)amide, 1 M in THF (0.798 mL, 0.798 mmol), then added to the reaction mixture. The reaction mixture was stirred at 50°C for 30 minutes. After cooling to room temperature, the reaction mixture was partitioned between dichioromethane (20 mL) and saturated aqueous ammonium chloride (20 mL). After separation, the organic extract was dried using a hydrophobic frit and evaporated to dryness. The residue was purified by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (89 mg, 0.169 mmol, 64 %). LCMS (Method D): Rt 1.36 minutes; m/z 526 ( H+).
Intermediate 109:
ho!inylmethyl)-2-pyridinyl]-1 ,3-benzothiazol-2-amine
Figure imgf000142_0001
Sodium ferf-butoxide (327 mg, 3.40 mmol) was suspended in tetrahydrofuran (5 mL) and stirred at 0°C. 6-Chloro-4-(4-morpholinylmethyi)-2-pyrid!namine (250 mg, 1.098 mmol) and 2-ch!oro-1 ,3-benzothiazole (224 mg, 1.318 mmo!) were added and the reaction was stirred at 0°C for 1 h and the allowed to warm up to room temperature while stirring for 16 hours. The reaction mixture was partitioned between dichloromethane (30 mL) and saturated ammonium chloride (30 mL). After separation, the organic extract was dried using a hydrophobic frit and evaporated to dryness. The residue was purified by chromatography on silica using a gradient elution from 0 to 100 % ethyl acetate in dichloromethane to afford the title compound (270 mg, 0.748 mmol, 68 %) as a brown solid. LCMS (Method D): Rt 1.23 minutes; m/z 361 (MH+).
Intermediate 110:
1 ,1 -dimethylethyl (3S)-3-{[6-(1 ,3-benzothiazol-2-ylamino)-4-(4-morpholinyl methyl)-2- pyridinyl]amino}-1-pyrroiidinecarboxylate
Figure imgf000142_0002
A round bottom flask was charged with A/-[6-chloro-4-(4-morpho!inylmethyl)-2-pyridinyl]- 1 ,3-benzothiazo!-2-amine (240 mg, 0.665 mmol), 1 , 1 -dimethylethyl (3S)-3-amino-1 - pyrrolidinecarboxyiate (136 mg, 0.732 mmol) and tetrahydrofuran (2 mL). The system was sealed and placed under an atmosphere of nitrogen using a vacuum purge (3 times) then heated to 50 °C. Lithium bis(trimethylsilyl)amide, 1 M in tetrahydrofuran (2 mL, 2.0 mmol) was added to {I .S-bis^.e-bisfl -methylethy phenylj^-imidazolidinyl }(chloro)(2-methyl-2- propen-1-yl)pal!adium (117 mg, 0.200 mmol) and the yellow solution was added to the round bottom flask. The reaction was heated at 50 °C for 1 hour. The reaction was cooled to room temperature and partitioned between dichloromethane (30 mL) and saturated ammonium chloride solution (30 mL). After separation, the organic extract was dried using a hydrophobic frit evaporated to dryness. The residue was purified by chromatography on silica using a gradient elution from 0 to 100 % ethyl acetate in dichloromethane to afford the title compound (202 mg, 0.396 mmol, 60 %) as a brown solid. LCMS (Method D): Rt 1.21 minutes; m/z 51 1 (MH+)- Intermediate 111 :
W-[6-chloro-4-{4-morpholinylmethyl)-2-pyridinyl]-6-(methyloxy}-1 ,3-benzoth
Figure imgf000143_0001
Sodium ierf-butoxide (327 mg, 3.40 mmol) was suspended in tetrahydrofuran (5 mL) and stirred at room temperature. 6-Chioro-4-(4-morpholinylmethyl)-2-pyridinamine (250 mg, 1.098 mmoi) and 2~chloro-6-(methyloxy)-1 ,3-benzothiazole (263 mg, 1 .318 mmoi) were added and the reaction was stirred at room temperature for 16 hours. A further portion of sodium fe/f-butoxide (110 mg, 1.14mmol) was then added and the reaction mixture left stirring at room temperature for 72 hours. The reaction was then heated at 60 °C for 6 hours. The reaction mixture was partitioned between dichloromethane (30 mL) and saturated ammonium chloride solution (30 mL). After separation, the organic extract was dried using a hydrophobic frit and evaporated to dryness. The residue was purified by chromatography on silica using a gradient elution from 0 to 100 % ethyl acetate in dichloromethane to afford the title compound (190 mg, 0.486 mmol, 44 %) as a white solid. LCMS (Method D): Rt 1.21 minutes; m/z 391 (MH+).
Intermediate 112:
1 ,1 -dimethylethyl (3S)-3-{[6-{[6-(methyloxy)-1 ,3-benzothiazol-2-yl]amino}-4-(4- morpholinylmethyl)-2-pyridinyl]amino}-1-pyrrolidinecarboxylate
Figure imgf000144_0001
A round bottom flask was charged with A/-[6-chioro-4-(4-morpholinylmethyl)-2-pyridinyi]-6- (methyloxy)-1 ,3-benzothiazol-2-amine(100 mg, 0.256 mmol), 1 ,1 -dimethylethyl (3S)-3- amino-1 -pyrrolidinecarboxylate (52 mg, 0.279 mmol) and tetrahydrofuran (2 ml_). The system was sealed and placed under an atmosphere of nitrogen using a vacuum purge (3 times) and heated to 50 °C. Lithium bis(trimethylsilyi)amide, 1 in tetrahydrofuran (1 mL, 1 .00 mmol) was added to {1 ,3-bis[2,6-bis(1 -methylethyl)phenyl]-2- imidazoiidinyl}(chloro)(2-methyl-2-propen-1 -yl)palladium (50 mg, 0.085 mmo!) and the yellow solution was added to the round bottom f!ask. The reaction was heated at 50 °C for 1 hour. The reaction mixture was cooled to room temperature and partitioned between dichloromethane (20 mL) and saturated ammonium chloride solution (20 mL). After separation, the organic extract was dried using a hydrophobic frit and evaporated to dryness. The residue was purified by chromatography on si!ica using a gradient elution from 0 to 100 % ethyl acetate in dichloromethane to afford the title compound (97 mg, 0.179 mmol, 70 %). LCMS (Method D): Rt 1.19 minutes; m/z 541 (MH+).
Intermediate 113:
W-[6-chloro-4-({[(1 S)-1 ,2,2-trimet^
benzothiazol-2-
Figure imgf000144_0002
A round bottom flask was charged with 6-chioro-4-({[(1 S)-1 ,2,2- trimethylpropyl]amino}methyl)-2-pyridinamine (150 mg, 0.620 mmol) and 2-chloro-1 ,3- benzothiazole (1 16 mg, 0.682 mmol) in dry A/,A/-dimethylformamide (3 mL) and stirred at room temperature under an atmosphere of nitrogen. Sodium hydride, 60 % w/w dispersion in mineral oil (44,7 mg, 1.861 mmol) was then added and the mixture stirred for 20 hours at room temperature under an atmosphere of nitrogen. A further portion of sodium hydride, 60 % w/w dispersion in mineral oil (44.7 mg, 1.861 mmol) was added and the reaction mixture stirred at room temperature for a further 4 hours. The reaction mixture was partitioned between dichioromethane (30 mL) and saturated ammonium chioride solution (30 mL). After separation, the organic extract was dried using a hydrophobic frit and evaporated to dryness. The residue was purified by chromatography on silica using a gradient eiution from 0 to 100% ethyl acetate in dichloromethane to afford title compound (137 mg, 0.365 mmoS, 59 %) which was used directly without further purification. LCMS (Method D): Rt 1.56 minutes; m/z 375 (MH+).
Intermediate 114:
1 ,1 -dimethylethyl (3S)-3-{[6-{1,3-benzothiazol-2-ylamino)-4-({[(1 S)-1 ,2,2-tri methylpropyl]amino}methyl)-2-pyridinyl]arnino}-1 -pyrrolidinecarboxyiate
Figure imgf000145_0001
A round bottom flask was charged with W-[6-chloro-4-({[(1 S)-1 ,2,2- trimethylpropyi]amino}methyl)-2-pyridinyl]-1 ,3-benzothiazo!-2-amine (137 mg, 0.365 mmol), 1 , 1-dimethylethyl (3S)-3-amino-1 -pyrrolidinecarboxylate (75 mg, 0.403 mmol) and tetrahydrofuran (2 mL). The system was sealed and placed under an atmosphere of nitrogen using a vacuum purge (3 times) and heated to 50 °C. Lithium bis(trimethyisilyl)amide, 1 M in tetrahydrofuran (1.1 mL, 1.10 mmol) was added to {1 ,3- bis[2,6-bis(1 -methylethyl)phenyl]-2Hmidazolidinyi(chloro)(2-methyl-2-propen-1 - yl)palladium (65 mg, 0.1 10 mmol) and the yeliow solution was added to the round bottom flask. The reaction was heated at 50 °C for 1 hour. The reaction was cooled to room temperature and partitioned between dichloromethane (30 mL) and saturated ammonium chloride solution (30 mL). After separation, the organic extract was dried using a hydrophobic frit and evaporated to dryness. The residue was purified by chromatography on silica using a gradient eiution from 0 to 100% ethyl acetate in dichloromethane to give the title compound (83 mg, 0.158 mmol, 43 %) as a brown solid. LCMS (Method D): Rt 1.47 minutes; m/z 525 (MH+).
Intermediate 115:
W-[6-chloro-4-({[(1S)-1 ,2,2-trimethylpropyl]amino}methyl)-2-pyridinyl]-6-(methyloxy)- -benzothiazol-2-amine
Figure imgf000146_0001
A round bottom flask was charged with 6-chloro-4-({[(1 S)-1 ,2,2- trimethylpropyl]amino}methyl)-2-pyridinamine (150 mg, 0.620 mmol) and 2-chloro-6- (methyioxy)-1 ,3-benzothiazole (136 mg, 0.682 mmol) in dry /V,A/-dimethylformamide (3 mL) and stirred at room temperature under an atmosphere of nitrogen. Sodium hydride, 60 % w/w dispersion in mineral oil (44.7 mg, 1.861 mmol) was then added and the mixture stirred for 20 hours at room temperature under an atmosphere of nitrogen. A further portion of sodium hydride, 60 % w/w dispersion in mineral oil (44,7 mg, 1.861 mmol) was added and the reaction mixture stirred at room temperature for a further 4 hours. The reaction mixture was partitioned between dichloromethane (30 mL) and saturated ammonium chloride solution (30 mL). After separation, the organic extract was dried using a hydrophobic frit and evaporated to dryness. The residue was purified by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (1 12 mg, 0.277 mmol, 45 %) as a grey solid. LC S (Method D); Rt 1.54 minutes; m/z 405 (MH+).
Intermediate 116:
1 ,1 -dimethylethyl (3S)-3-{[6-{[6-(methyloxy)-1 ,3-benzothiazol-2-yl]amino}-4-({[(1 S)- 1 ,2,2 rimethylpropyl]amino}methyl)-2^yridinyl]amino}-1-pyrro!idinecarboxy
Figure imgf000146_0002
A round bottom flask was charged with W-[6-ch!oro-4-({[(1 S)-1 ,2,2- trimethylpropyl]amino}methyl)-2-pyridinyl3-6-(methyloxy)-1 ,3-benzothiazol-2-amine (1 12 mg, 0.277 mmol), 1 , 1 -dimethylethyl (3S)-3-amino-1-pyrro!idinecarboxylate (57 mg, 0.306 mmo!) and tetrahydrofuran (2 mL). The system was sealed and placed under an atmosphere of nitrogen using a vacuum purge (3 times) and heated to 50 °C. Lithium bis(trimethylsilyl)amide, 1 M in tetrahydrofuran (0.9 mL, 0.90 mmol) was added to {1 ,3- bis[2,6-bis(1 -methylethyl)phenyl]-2-imidazoiidinyl}(chloro)(2-methyl-2-propen-1 - yl)palladium (50 mg, 0.085 mmol) and the yellow solution was added to the round bottom flask. The reaction was heated at 50 °C for 1 hour. The reaction was cooled to room temperature and partitioned between dichioromethane (30 mL) and saturated ammonium chloride solution (30 mL). After separation, the organic extract was dried using a hydrophobic frit and evaporated to dryness. The residue was purified by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (37 mg, 0.0067 mmol, 24 %) as a brown glass. LCMS (Method D): Rt 1.45 minutes; m/z 555 (MH+).
Intermediate 117:
2-amino-6-chloro-W-t(1R)-1 ,2,2 rimethyipropyl]-4-pyridinecarboxamide
Figure imgf000147_0001
To a solution of 2-amino-6-chioroisonicotinic acid (1 g, 5.79 mmol) in N,N- dimethylformamide (20 mL) stirred at room temperature was added 1 ,1 '- carbonyldiimidazole (1.150 g, 7.09 mmol) in one charge. The reaction mixture was stirred at room temperature for 1 hour under an atmosphere of nitrogen. [(1 f?)-1 ,2,2- trimethylpropyl]amine (0.704 g, 6.95 mmol) was then added and the reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was partitioned between dichioromethane (20 mL) and saturated aqueous sodium carbonate (20 mL). After separation the organic extract was washed with saturated aqueous sodium carbonate (20 mL), dried using a hydrophobic frit and evaporated to dryness to give the crude product as an off-white solid. The residue was purified by chromatography on silica using a gradient elution from 0 to 50 % ethyl acetate in cyciohexane to afford the title compound (1 g, 3.91 mmol, 67 %) as a white solid. LCMS (Method D): Rt 0.97 minutes; m/z 256 (MH+).
Intermediate 118:
6-chloro-4-({[(1 R)-1,2,2-trimethylpropyl]amirto}methyi)-2-pyridinamine
Figure imgf000148_0001
To a solution of 2-amino-6-chloro-A/-[(1 R)-1 ,2,2-trimethylpropyl]-4-pyridine carboxamide (1.3 g, 5.08 mmol) in dichloromethane (10 mL) stirred at room temperature was added 1 M borane-tetrahydrofuran complex (20.33 mL, 20.33 mmol) dropwise. The reaction mixture was stirred at room temperature under a nitrogen atmosphere for 72 hours then heated at 40 °C for 2 hours. A further equivalent of 1 borane-tetrahydrofuran complex (5.08 mL, 5.08 mmol) was added and the reaction mixture was stirred at 40 °C for 3 hours and then at 50 °C for 2 hours. The solution was cooled to 0°C before addition of 2M aqueous HCI (50 mL). Stirring was continued at 0 °C for 1 hour. 10M aqueous sodium hyroxide was carefully added to adjust the pH to 12. The product was extracted with dichloromethane (2 x 100 mL). The organic extracts were combined, dried using a hydrophobic frit and evaporated to dryness to afford an orange oil. The residue was dissolved in dichloromethane (20 mL) and washed with saturated aqueous ammonium chloride (20 mL). The aqueous phase was extracted with dichloromethane (50 mL). The organic extracts were combined, dried using a hydrophobic frit and evaporated to dryness to give the title compound (1.16 g, 4.80 mmol, 94 %) as orange crystals. LCMS (Method D): Rt 1.13 minutes; m/z 242 (MH+).
Intermediate 119:
W-[6-chloro-4-({[(1 R)-1 ,2,2-trimethyIpropyl]amino}methyl)-2-pyridinyi3
1 ,3]thiazoio[5,4- i]pyridin-2-amine
Figure imgf000148_0002
A round bottom flask was charged with 6-chloro-4-({[(1 )-1 ,2,2-trimethyi propyl]amino}methyl)-2-pyridinamine (1 g, 4.14 mmol) and 2-bromo[1 ,3]thiazolo[5,4- b]pyridine (0.925 g, 4.30 mmol) in dry A/,W-dimethylformamide (30 mL), placed under an atmosphere of nitrogen and stirred at room temperature. Sodium hydride, 60 % w/w dispersion in mineral oil (0.496 g, 12.41 mmol) was then added and the mixture stirred under a nitrogen atmosphere for 4 hours at room temperature. The reaction mixture was partitioned between dichloromethane (100 mL) and saturated ammonium chloride solution {100 mL). After separation, the organic extract was dried using a hydrophobic frit and evaporated to dryness to give a brown solid. The residue was triturated with acetonitriie (50 mL). The resulting solid was filtered through a Buchner funnel, washed with acetonitriie (10 mL), and collected to give the title compound (1.06 g, 2.82 mmol, 68 % yield) as an off-white solid. LC S (Method D): Rt 1.40 minutes; m/z 376 (MH+).
Intermediate 120:
1,1-dimethylethyl (S^-S-iie-iEl^lthiazoiotS^-iJlpyridin-a-ylaminoJ^-liaiRJ-l ^^- trimeth lpropyl]amino}methy!)-2-pyridinyl]amino}-1-pyrrolidine carboxylate
Figure imgf000149_0001
A microwave vial was charged with A/-[6-chloro-4-({[( R)-1 ,2,2- trimethy!propyl]amino}methyi)-2-pyridinyl] [1 ,3]thiazo!o[5,4-b]pyridtn-2-amine
(250 mg, 0.665 mmol) and 1 , 1 -dimethy!ethyl (3R)-3-amino-1-pyrrolidinecarboxylate (186 mg, 0.998 mmol), dissolved in tetrahydrofuran (1 mL). The system was sealed and placed under an atmosphere of nitrogen using a vacuum purge (3 times). {1 ,3-Bis[2,6-bis(1- methyl ethyl)phenyl]-2-imidazolidinyi}(chloro)(2-methyl-2-propen-1 -yi) pa!ladium (1 17 mg, 0.200 mmol) was mixed with lithium bis(trimethylsilyl)amine, 1 M in tetrahydrofuran (1.995 mL, 1.995 mmo!), then added to the reaction mixture. The reaction mixture was stirred at 50 °C for 30 minutes. The reaction was cooled to room temperature. The reaction mixture was partitioned between dichloromethane (20 mL) and saturated aqueous ammonium chloride (20 mL). After separation, the organic extract was dried using a hydrophobic frit and evaporated to dryness. The residue was triturated with acetonitriie (50 mL). The resulting solid was filtered through a Buchner funnel, washed with acetonitriie (10 mL), and collected to give the title compound (262 mg, 0,498 mmol, 75 % yield) as an off-white solid. LCMS (Method D): Rt 1.36 minutes; m/z 526 (MH+).
Intermediate 121 : -amino-6-chloro-W-[(1S)-1 ,2,2-trimethyIpropyl]-4-pyridinecarboxamide
Figure imgf000150_0001
To a solution of 2-amino-6-chloroisonicotinic acid (52 g, 301 mmol) in N,N- dimethylformamide (200 mL) stirred at room temperature was added 1 , 1 '- carbonyldiimidazole (58.6 g, 362 mmol) in one charge. The reaction mixture was stirred at room temperature for 60 minutes. [(1 S)-1 ,2,2-trimethylpropyl]amine (39.3 mL, 362 mmol) was then added and the reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was partitioned between dichloromethane (500 mL) and saturated aqueous sodium carbonate (500 mL). After separation the organic extract was washed further with saturated aqueous sodium carbonate (200 mL), dried using a hydrophobic frit and concentrated under reduced pressure to afford title compound (61.5 g, 240 mmol, 80 %) as a white solid. LCMS (Method D): Rt 0.97 minutes; m/z 256 (MH+).
Intermediate 122:
e-chl ro-^i^lSJ-l^^-trimethylpropyljaminoJmethy ^-pyridinamine
Figure imgf000150_0002
To a solution of 2-amino-6-chloro-A/-[(1 S)-1 ,2,2-trimethylpropyl]-4-pyridinecarboxamide (3.29 g, 12.86 mmol) in dichloromethane (20 mL) stirred at room temperature was added borane in tetrahydrofuran (1 M, 64.3 mL, 64.3 mmol) dropwise. The reaction mixture was stirred at room temperature for 16 hours under an atmosphere of nitrogen. The reaction was then heated under reflux conditions (50 °C) for 7 hours. The reaction was cooled down to room temperature. 5M aqueous sodium hydroxide solution (50 mL) was carefully added (careful, effervescence) and the reaction was stirred at room temperature for 1 hour. The product was extracted with dichloromethane (2 x 150 mL). The organic extracts were combined, dried with magnesium sulfate, filtered and evaporated under reduced pressure to afford a yellow oil (4.9 g). The oil was partitioned between dichloromethane (50 mL) and saturated ammonium chloride solution (30 mL). After separation, the organic extract was dried using a hydrophobic frit and evaporated to dryness to give the title compound as an off-white solid (2.6 g, 10.75 mmol, 84 %). LCMS (Method D): Rt 1.15 minutes; m/z 242 (MH+).
Intermediate 123:
W-[6-chloro-4-({t(1S)-1,2,2-trimethylpropyl]amino}methyl)-2-pyridinyl]
3]thiazoio[5,4-bJpyridin-2-amine
Figure imgf000151_0001
To a soiution of sodium fe -butoxide (35.0 g, 364 mmol) in tetrahydrofuran (250 mL) was added a solution of 6-chloro-4-({[(1 S)-1 ,2,2-trimethylpropyl]amino}methyl)-2-pyridinamine (28.4 g, 117 mmol) and 2-bromothiazolo[5,4-6]pyridine (27.8 g, 129 mmol) in tetrahydrofuran (300 mL) at such a rate that the temperature remained below 10 °C. The reaction was then allowed to warm to room temperature over one hour. Saturated ammonium chloride (200 mL) and water (50 mL) were added and the aqueous extracted with 2-methyltetrahydrofuran (2 x 50mL). The combined organics were dried over magnesium sulfate, filtered and concentrated under reduced pressure to give an orange solid. This was triturated with diethyl ether (200 mL) and the resultant precipitate collected by filtration, washed with diethyl ether and dried under vacuum to give the title compound (29.8 g, 79.4 mmol, 67.5 %). LCMS (Method A): Rt 0.76 minutes; m/z 376 (MH+).
Intermediate 124:
1 , -dimethyiethyl {3S)-3-{[6-([1 ,3]thiazolo[5,4-i ]pyridin-2-ylamino)-4-({[(1 S)-1 ,2,2- rimethylpropyl]amino}methyl)-2-pyridinyl]amino}-1 -pyrro!idinecarboxylate
Figure imgf000151_0002
To a stirred solution of 1 ,1 -dimethylethyl (3S)-3-amino-1-pyrrolidinecarboxylate (4.34 mL, 24.87 mmol) and A/-[6-chloro-4-({[(1 S)-1 ,2,2-trimethySpropyl]amino}methyl)-2-pyridinyl] [1 ,3]thiazolo[5,4-£>]pyridin-2-amine (8.5 g, 22.61 mmol) in tetrahydrofuran (60 mL) at 62-65 °C was added a solution of {1 ,3-bis[2,6-bis(1 -methylethyl)pheny!]-2- imidazolidinyl}(chloro)(2-methyi-2-propen-1-yi)palladium (3.99 g, 6.78 mmol) in lithium bis(trimethylsilyl)amide, 1 M in tetrahydrofuran (67.8 mL, 67.8 mmol) portionwise, ensuring that the temperature remained 60-65 °C. After the addition was complete the reaction was allowed to stir for 30 minutes. The reaction was cooled to room temperature. Water (20 mL) was added and the aqueous phase was extracted with 2-methyltetrahydrofuran (2 x 20 mL). The combined organics were dried over magnesium sulfate, filtered and concentrated to a gum. Trituration with dichloromethane afforded an off-white solid which was collected by filtration, washed with dichloromethane and dried to give the title compound (6.5 g, 12.4 mmol, 55 %). LCMS (Method A): Rt 0.93 minutes; m/z 526 (MH+).
Intermediate 125:
/V-ftSSJ-S-pyrrolidinyil-W'-il .SJthiazolo^^-bl yridin-a-yl^-iiKISJ-l ^^- trimethylpropyl]amino}methyl)-2,6-pyridinediamine, trihydrochloride
Figure imgf000152_0001
1 , 1-Dimethylethyl (3S)-3-{[6-([1 ,3]thiazolo[5,4-/)3pyridin-2-ylamino)-4-({E(1 S)-1 ,2,2-trimethyl propyl]amino}methyl)-2-pyridinyl]amino}-1-pyrrolidinecarboxylate (45 g, 86 mmol) was dissolved in dichloromethane (300 mL) and methanol (300 mL) and treated with HCI, 4M in dioxane (400 mL). After stirring at room temperature for 2 hours, the solid was filtered and washed with dichloromethane. The yeliow solid was dried under vacuum to give the title compound (39 g, 72.9 mmol, 85 %). LCMS (Method A): Rt 0.55 minutes; m/z 426 (MH+).
Intermediate 126:
W-IISSJ-S-pyrroiidinyll-W-Il^lthiazolo^^p ridin^- l^-tiKI SJ-l^^- trimethylpropyl]amino}methyl)-2,6-pyridinediamine
Figure imgf000152_0002
W-[(3S)-3-Pyrrolidinyl]-W-[1 ,3]thiazolo[5,4-6]pyridin-2-yl-4-({[(1 S)-1 ,2,2-trimethyl propyl]amino}methyi)-2,6-pyridinediamine trihydroch!oride (5 g, 9.35 mmoS) was dissolved in dichloromethane (50 mL) and methanol (50 mL) and treated with MP-carbonate resin (20 g, 56 mmol). After stirring at room temperature for 3 hours the resin was filtered off and washed with dichloromethane/methanoi (1 : 1 ). The filtrate was evaporated to dryness to give the title compound (4 g, 9.4 mmol, 101 % of the theoretical yield). LC S (Method E): Rt 0.57 minutes; m/z 426 (MH+).
Intermediate 127:
5- methyloxy)[1 ,3]thiazolo[5,4-J ]pyridin-2-amine
Figure imgf000153_0001
Potassium thiocyanate (100 g, 1029 mmol) was dissolved in acetic acid (500 mL), cooled to 0 °C in an ice bath and 6-methoxypyridin-3-amine (25 g, 201 mmol) dissolved in acetic acid (50 mL) added over 30 minutes. Bromine (12.5 ml, 243 mmol) dissolved in acetic acid (50 mL) wad added over 20 minutes. The reaction was stirred at 0 °C for 2 hours and was warmed to room temperature overnight. The slurry was filtered and the cake was washed with acetic acid (300 mL). The solid was suspended in ethyl acetate (500 mL) and saturated aqueous sodium bicarbonate (400 mL) was added slowly. After a large carbon dioxide evolution, the slurry was neutralised with 10M aqueous sodium hydroxide to reach pH 7. The slurry was filtered and the filtrate layers separated. The organic layer was washed with saturated aqueous sodium bicarbonate (250 mL), dried using a hydrophobic frit and evaporated under reduced pressure to afford the title compound (20 g, 1 1 1 mmol, 55 % yield) as a red solid. LCMS (Method E): Rt 0.48 minutes; m/z 182 (MH+). tntermediate 128:
2-bromo-5- methyloxy)[1 ,3]thiazolo[5,4-/)]pyridine
Figure imgf000153_0002
5-(Methyloxy)[1 ,3]thiazolo[5,4-/b]pyridin-2-amine (5 g, 27.6 mmol), tosic acid (6.30 g, 33.1 mmol), sodium nitrite (2.284 g, 33.1 mmol) and tetrabutylammonium bromide (17.79 g, 55.2 mmol) were dissolved in acetonitrile (120 mL). Copper(ll) bromide (0.062 g, 0.276 mmol) was added and the reaction was stirred under nitrogen for 72 hours at room temperature. The reaction mixture was concentrated under reduced pressure to give a red / brown sludge, which was dissolved in dichloromethane (250 mL) and washed with water (50 mL), The layers were separated using a hydrophobic frit and the dichloromethane layer evaporated under reduced pressure to give a red /brown sludge. Acetonitrile (50 mL) was added and the slurry stirred for two hours. The slurry was filtered, the cake washed with acetonitrile (2 x 25 mL) and dried under vacuum to give the title compound (1 g, 4.1 mmol, 15% yield) as red solid. The filtrate was concentrated. The suspension was cooled to 0 °C, stirred for 1 hour and filtered. The cake was washed with acetonitrile (10 mL) and dried under vacuum to afford the title compound (5 g, 20,4 mmo!, 74% yield) as a dark red solid. LCMS (Method E): Rt 1.09 minutes; 1 H NMR (400 MHz, DMSO-d6) d 8.29 (d, J = 9.0 Hz, 1 H), 7.02 (d, J = 9.0 Hz, 1 H), 3.94 (s, 3H). Intermediate 129:
W-(6-chloro-4-{[c/s-2,6-dimethyl-4-morpholinyl3methyl}-2-pyndinyl)-5- {methyloxy)[1 ,3]thiazolo[5,4-i)]pyridin-2-amine
Figure imgf000154_0001
Sodium terf-butoxide (513 mg, 5.33 mmol) was dissolved in tetrahydrofuran (5 mL) and cooled to 0 °C. 6-Chloro-4-{[c/s-2,6-dimethyl-4-morpholinyl]methyl}-2-pyridinamine (440 mg, 1.720 mmol) and 2-bromo-5-(methyloxy)[1 ,3]thiazolo[5,4-b]pyridine (464 mg, 1.893 mmol) dissolved in tetrahydrofuran (5 mL) were added dropwise and the reaction stirred at 0 °C for 15 minutes and allowed to warm to room temperature. The reaction was stirred for 1 hour. Aqueous saturated ammonium chloride (7.5 mL) and dichloromethane (12 mL) were added, the biphasic solution was stirred for 20 minutes and the layers separated using a hydrophobic frit. The aqueous layer was extracted with dichloromethane (10 mL) and the layers separated using a hydrophobic frit. The combined dichloromethane layers were evaporated under reduced pressure to give a brown solid. Dichloromethane (10 mL) and methanol (10 mL) were added to form a slurry and the dichloromethane evaporated under reduced pressure to leave the solid in methanol. The slurry was stirred for 30 minutes and filtered. The cake was washed with methanol (3 x 10 mL) and dried under vacuum for 1 hour to give the title compound as a white solid (450 mg, 1.07 mmol, 62 % yield). The combined mother liquors were evaporated under reduced pressure to give a concentrated solution (~5 mL). The slurry was left to crystallise overnight, was filtered, washed with methanol (5 mL) and the solid dried under vacuum to afford the title compound (70 mg, 0.17 mmol, 10 % yield) as an off white solid. LCMS (Method E): Rt 0.87 minutes; m/z 420 (MH+). Intermediate 130:
W-{6-chloro-4-[{dimethylamino)methylJ-2-pyrtdm^
Jt> ridin-2-amine
Figure imgf000155_0001
Sodium ferf-butoxide (287 mg, 2.99 mmol) was dissolved in tetrahydrofuran (2 ml_) and cooled to 0 °C. 6-Chloro-4-((dimethyiamino)methyl)pyridin-2-amine (185 mg, 0.996 mmol) and 2-bromo-5-(methy!oxy)[1 l3]thiazo!o[5,4-b]pyridine (269 mg, 1.096 mmol) dissolved in tetrahydrofuran (2 mL) were added dropwise and the reaction stirred at 0 °C for 30 minutes. The reaction was warmed to room temperature and stirred for 3 hours. Further sodium terf-butoxide (100 mg, 1.04 mmoi) was added and the reaction stirred for 30 minutes. Saturated aqueous ammonium chloride (4 mL) was added, the mixture stirred for 10 minutes and dichloromethane (25 mL) added. The layers were separated using a hydrophobic frit. The aqueous layer was extracted with dichloromethane (2 x 10 mL) and the layers separated using hydrophobic frits. The combined organic layers were evaporated under reduced pressure to give a brown solid. The solid was purified by chromatography on silica using a gradient e!ution from 0 to 25 % methanol in dichloromethane to afford the title compound (237 mg, 0.677 mmo!, 68 % yield) as a yellow solid. LC S (Method E); Rt 0.79 minutes; m/z 350 (MH+). Intermediate 131 :
1 ,1 -dimethylethy! (3S)-3-[(4-[(dimethylamino)methyl]-6-{[5-
(methyloxy)[1 ,3]thiazolo[5,4- j3pyridin-2-yl]amino}-2-pyridinyl)amino]-1 - rrolidinecarbox late
Figure imgf000155_0002
W-{6-Chloro-4-[(dimethylamino)methyl]-2-pyridinyl}-5-(methyloxy)[1 ,3]thiazolo[5,4- £]pyridin-2-amine (178 mg, 0.509 mmoi) was weighed into a microwave reactor via! and 1 ,1 -dimethylethy! (3S)-3-amino-1-pyrrolidinecarboxylate (0.098 mL, 0.560 mmoi) dissolved in anhydrous tetrahydrofuran (1 mL) added. The system was sealed and placed under an atmosphere of nitrogen using a vacuum purge and heated to refiux. {1 ,3-Bis[2,6- bis(1-methylethyl)phenyl]-2-imidazolidinyl}(chloro)(2-methyl-2-propen-1 -yl)paliadium (90 mg, 0.153 mmo!) dissolved in lithium bis(trimethyisilyl)amide, 1 in tetrahydrofuran (1.52 mL, 1.52 mmol) was added in three equal portions over 15 mintues. The reaction was heated to reflux for 30 minutes. Further {1 ,3-bis[2,6-bis(1-methylethyl)phenyl]-2- imidazoiidinyl}(chloro)(2-methyl-2-propen-1 -yl)paliadium (45 mg, 0.076 mmol) dissolved in lithium bis(trimethySsilyi)amide, 1 M in tetrahydrofuran (0.76 mL, 0.76 mmol) was added in three equal portions over 15 mintues and the reaction heated to reflux for 30 minutes. Water (5.0 mL) was added and the reaction cooled to room temperature. 2- Methyltetrahydrofuran (5 mL) was added and the layers were separated. The aqueous layer was extracted with 2-methyltetrahydrofuran (2 x 10 mL). The combined layers were dried with anhydrous magnesium sulfate and evaporated under reduced pressure to give a brown oil. Diethyl ether (4 mL) was added to the oil and the yeliow solid was stirred for 5 minutes. Further diethyl ether (4 mL) was added, the slurry stirred for 20 minutes and filtered to afford the title compound (148 mg, 0.296 mmol, 58 % yield) as a yellow solid. LCMS (Method A): Rt 0.95 minutes; m/z 500 (MH+).
Intermediate 132:
W-[6-chloro-4-({[(1 S)-1 ,2,2 rimethylpropyl]amino}methyl)-2-pyridinyl]-5- (meth loxy)[1,3]thiazolo[5,4-f)]pyridin»2-amine
Figure imgf000156_0001
Sodium ierf-butoxide (616 mg, 6.41 mmol) was dissolved in tetrahydrofuran (5 mL) and cooled to 0 °C. 6-Chloro-4-({[(1 S)-1 ,2,2-trimethylpropyl]amino}methyl)-2-pyridinamine (500 mg, 2.068 mmol) and 2-bromo-5-(methyloxy)[1 ,3]thiazoio[5,4-b]pyridine (558 mg, 2.275 mmol) dissolved in tetrahydrofuran (5 mL) were added dropwise and the reaction stirred at 0 °C for 15 minutes and then allowed to warm to room temperature. The reaction was stirred for 2 hours. Saturated aqueous ammonium chloride (7.5 mL) and dichioromethane (12 mL) were added, the biphasic solution was stirred for 1 hour and the layers separated using a hydrophobic frit, The dichioromethane layer was evaporated under reduced pressure to give a brown oil. The oil was purified by chromatography on silica using a gradient elution from 0 to 25 % methanoi in dichioromethane to afford a black solid. The solid was suspended in methanoi (25 mL) and the solvent was evaporated under reduced pressure to give a slurry of approximately 6 mL. The slurry was cooled to room temperature, stirred for 2 hours, filtered and the cake washed with methanol (5 mL). The solid was dried under vacuum at 40 °C to afford the title compound (320 mg, 0.788 mmol, 38% yield) as a white solid. LCMS (Method E): Rt 0.93 minutes; m/z 406(MH+).
Exam le 1 :
W-(2-{[4-[(methyloxy)methyl]-6-([1,3]thiazolo[5,4-Jb]pyridin-2-ylamino)-2- pyrimidinyl3amino}ethyl)-2-propenamide
Figure imgf000157_0001
A mixture of /V2-(2-aminoethyl)-6-[(methyloxy)methyl3-W4-[1 ,3]thiazolo[5,4-i)]pyridin-2-yl- 2,4-pyrimidinediamine hydrochloride (70 mg, 0.190 mmol), A/./V-diisopropyiethyiamine (0.199 mL, 1.142 mmol) and 2-propenoyl chloride (0.022 mL, 0.266 mmol) in /V-methyl-2- pyrrolidone (2 mL) was stirred at room temperature for 72 hours. The residue was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (33 mg, 0.086 mmol, 45 % yield). LCMS (Method B): Rt 1.87 minutes; m/z 386 (MH+).
Example 2:
W-[2-({4 (6-bromo-1,3-benzothiazoI-2-yi)aminoj-2-pyrimidinyl}amino)ethyl]-2- propenamide
Figure imgf000157_0002
A mixture of W2-(2-aminoethyl)-W*-(6-bromo-1 ,3-benzothiazol-2-yl)-2,4-pyrimidinediamine (130 mg, 0.356 mmol), 2-propenoyl chloride (0.039 mL, 0.463 mmol) and N,N- diisopropylethyjamine (0.373 mL, 2.136 mmol) in tetrahydrofuran (2.5 mL) was stirred at room temperature for 2 hours. The white solid was filtered under vacuum and washed with ethyl acetate. The filtrate was evaporated to dryness. The residue was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (4.6 mg, 0.011 mmol, 3 % yield). LC S (Method B): Rt 2.39 minutes; m/z 419/421 ( H+). Example 3:
f rans-4-{[4-(4-m o r p h o I i n y I m ethy I ) -6-( ^ ^
pyrimidinyl]amino}cyclohexyl methanesulfonate
Figure imgf000158_0001
A mixture of frans-4-{[4-(4-morpho!inylmethyl)-6-([1 ,3]thiazolo[5,4-b]pyridin-2-ylamino)-2- pyrimidiny!]amino}cyclohexanol (which may be prepared as described in WO 2010/106016) (188 mg, 0.426 mmol), methanesulfonyl chloride (0.066 mL, 0.852 mmol) and Λ/,/V-diisopropylethy!amine (0.149 mL, 0.852 mmol) was stirred at room temperature in dichloromethane (5 mL) for 72 hours. The reaction mixture was partitioned between dichloromethane (30 mL) and saturated aqueous sodium carbonate (30 mL). The phases were separated, the organic extract was dried using a hydrophobic frit and evaporated to dryness. The residue was purified by chromatography on silica using a gradient elution from 0 to 15 % methanol (+1 % triethyjamine) in dichloromethane to afford the title compound (126 mg, 0.243 mmol, 57 % yield) as a yellow solid. LCMS (Method B): Rt 2.22 minutes; m/z 520 (MH+).
Example 4:
W -(1-acryloyl-3^iperidinyl}-6-[{methyloxy)methyl3^
-2,4-pyrimidinediamine
Figure imgf000158_0002
A microwave vial was charged with W-[6-[(methyloxy)methyl]-2-(methylsuifonyl)-4- pyrimidinyi][1 ,3]thiazolo[5,4-/)]pyridin-2-amine (500 mg, 1.423 mmol), 1 ,1-dimethylethyi 3- amino-1 -piperidinecarboxylate (570 mg, 2.85 mmol) and isopropanol (5 ml_). The microwave vial was sealed and was heated in a Biotage Initiator microwave at 150 °C for 1.5 hours. The residue was purified by chromatography on silica using a gradient elution from 0 to 15 % methanol (+1 % triethylamine) in dichloromethane. After evaporation of the desired fractions to dryness, the residue was dissolved in dichloromethane and treated with 4M HC! in dioxane (0.6 mL, 2.4 mmoi) at room temperature. The solution was stirred at room temperature overnight. The reaction mixture was filtered under reduced pressure, dried to give 185 mg of a solid. A mixture of the obtained solid (50 mg), 2-propenoyl chloride (0.015 ml, 0.175 mmol) and A/JV-diisopropylethyiamine (0.141 ml, 0.808 mmol) in /V-methyl-2-pyrroiidone (1 ml) was stirred at room temperature for 2 hours. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (12 mg, 0.028 mmol, 2 % yield). LCMS (Method C): Rt 1.54 minutes; m/z 426 (MH+). Example 5:
W2-(1 -acryloyl-4-piperidinyl)-6-[(methyloxy^
-2,4-pyrimidinediamine
Figure imgf000159_0001
A mixture of e-timethyloxyJmethyll-A/^-piperidi yl-A^-t SJthiazoiofS^-i jpyridi ^-yl^^- pyrimidinediamine hydrochloride (50 mg, 0.123 mmoi), 2-propenoy! chloride (0.015 ml, 0.175 mmol) and A/,A/-diisopropylethyiamine (0.141 ml, 0.808 mmo!) in /V-methyl-2- pyrrolidone (1 ml) was stirred at room temperature for 18 hours. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (13 mg, 0.030 mmol, 24 % yieid). LCMS (Method C): Rt 1.50 minutes; m/z 426 (MH+).
Example 6:
frans-4-{[4-[{methyloxy)methyl]-6-(i1 ,3]thiazolo[5,4-i)3pyridin-2-ylamino)-2- pyrimidinyl]amino}cyclohexyl methanesulfonate
Figure imgf000160_0001
A mixture of fra ?s-4-{[4-[(methyloxy)methyl3-6-([1 ,33thiazolo[5,4-j ]pyridin-2-ylamino)-2- pyrimidinyi]amino}cyclohexanol (26 mg, 0.067 mmol), methanesulfony! chloride (0.01 mL, 0.135 mmol) and A/,W-dissopropyiethylamine (0.023 mL, 0.135 mmol) was stirred at room temperature in dichloromethane (5 mL) for 18 hours. A second portion of methanesulfonyl chloride (0.001 mL, 0.135 mmol) and Λ/,/V-diisopropylethy!amine (0.023 mL, 0.135 mmol) was added and the reaction mixture was stirred for 18 hours. A third portion of methanesulfonyl chloride (0.261 mL, 3.36 mmol) and W,N-dtisopropylethylamine (0.587 mL, 3.36 mmol) was added and the reaction mixture was stirred for 3 hours. The reaction mixture was evaporated to dryness. The residue was subjected to purification by mass directed automated preparative HPLC (formic acid modifier). After concentration of the solvent, the residue was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (5 mg, 0.01 1 mmol, 16 % yield). LCMS (Method B): Rt 2.30 minutes; m/z 465 (MH+).
Example 7:
W-{4-{[4-[(methyfoxy)methyl]-6-([1 ,3]thiazolo[5,4- j]pyridin-2-y!amino)-2- pyrimidinyiJamino}butyl)-2-propenamide
Figure imgf000160_0002
A mixture of /V2-(4-aminobutyi)-6-[(methyloxy)methyl]-A/4-[1 ,3]thiazolo[5,4-i)]pyridin-2-yl- 2,4-pyrimidinediamine hydrochloride (50 mg, 0.126 mmol), 2-propenoyl chloride (0.015 mL, 0.181 mmol) and N,N-diisopropylethylamine (0.146 mL, 0.835 mmol) in W-methyl-2- pyrrolidone (2 mL) was stirred at room temperature for 2 hours. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (13 mg, 0.031 mmol, 25 % yield). LCMS (Method C): Rt 1.38 minutes; m/z 414 (MH+). Example 8:
W-(3-{[4-[{methyloxy)methyf]-6-([1,3]thiazoloE5,4-i)]pyridin-2-ylarnino)-2- pyrimidinyl]amino}propyl)-2-propenamide
Figure imgf000161_0001
A mixture of W2-(3-aminopropyl)-6-[(methyloxy)methyl]-A/4-[1 l3]thiazolo[5,4- j3pyridiri-2-yi- 2,4-pyrimidinediamine hydrochloride (50 mg, 0.131 mmol), /V,A/-diisopropylethylamine (0.152 mL, 0.869 mmol) and 2-propenoyl chloride (0.016 mL, 0.188 mmoi) in W-methyl-2- pyrroiidone (2 mL) was stirred at room temperature for 2 hours. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (13 mg, 0.032 mmol, 25 % yield). LCMS (Method B): Rt 1.91 minutes; m/z 400 (MH+).
Example 9:
W-(irans-2-{[4-[(methyloxy)methyl}-6-([1,3]thiazolo[5,4-£»]pyNdin-2-ylamino)-2- pyrimidinyl]amino}cyclohexyl)-2-propenamide
Figure imgf000161_0002
A mixture of ^-[(transJ^-aminocyclohexylj-e-KmethyioxyJmethylJ-A/^I .SjthiazoioiS^- J ]pyridin-2-yl-2,4-pyrimidinediamine (112 mg, 0.291 mmol), 2-propenoyl chloride (0.025 mL, 0.291 mmol) and W,W-diisopropyiethylamine (0.101 mL, 0.581 mmol) in /V-methyl-2- pyrrolidone (1 mL) was stirred at room temperature for 10 minutes. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (24 mg, 0.055 mmol, 19 % yield). LCMS (Method B): Rt 2.28 minutes; m/z 440 (MH+).
Example 10: W-(c/s-3-{[4-i(methyloxy)methyl]-6-([1,3]thiazolo[5,4-bJpyridin-2-ylami pyrimidinyl]amino}cycfohexyl)-2-propenamide
Figure imgf000162_0001
A mixture of A/2-[c/'s-3-aminocyclohexyl]-6-[(methyloxy)methyl]-A '-[1 ,33thiazo[o[5,4- b]pyridin-2-yl-2,4-pyrimidinediamine (73 mg, 0.189 mmol), 2-propenoyl chloride (0.021 mL, 0.246 mmo!) and Λ/JV-diisopropylethylarnine (0.198 mL, 1.136 mmol) in /V-methyl-2- pyrrolidone (1 mL) was stirred at room temperature for 10 minutes. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (26 mg, 0.059 mmol, 31 % yield). LC S (Method B): Rt 2.10 minutes; m/z 440 (MH+).
Example 11 :
W-(ira/7s-3-{[4-[(methyIoxy}methyl]-6-([1 ,3]thiazoIo[5,4-Jb]pyridin-2-ylamino)-2- pyrimidinyl]amino}cyclohexyl)-2-propenamide
Figure imgf000162_0002
A microwave vial was charged with W-[6-[(methyloxy)methyl]-2-(methylsulfonyl)-4- pyrimidinyl][1 ,3]thiazolo[5,4-i)]pyridin-2-amine (200 mg, 0.569 mmol), 1 ,3- cyclohexanediamine (97 mg, 0.854 mmol) and isopropanol (4 mL). The viai was sealed and was heated in a Biotage Initiator microwave at 150 °C for 30 minutes. After cooling, the reaction was filtered under reduced pressure and the solid was washed with isopropano!. The filtrate was evaporated to dryness to give a yellow solid. The solid was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford 29 mg of the intermediate. The intermediate (25 mg), 2- propenoyl chloride (0.007 mL, 0.084 mmol) and Λ/,/V-diisopropylethylamine (0.068 mL, 0.389 mmol) in A/-methyl-2-pyrrolidone (1 mL) was stirred at room temperature for 10 minutes. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound
0.01 1 mmol, 2 % yield). LCMS (Method B): Rt 2.15 minutes; m/z 440 (MH+).
Example 12:
W2-[{3S)-1 -acryloyl-3-pyrrolidinyl]-6-[(methyloxy)methyl]-W -[1,3]thiazolo[5,4- imidinediamine
Figure imgf000163_0001
A mixture of 6-[(methyloxy)methyl3-/V2-[(3S)-3-pyrrolidinyl]-/V4-[1 ,33thiazo[o[5,4-jb]pyridin-2- yf-2,4-pyrimidinediamine hydrochloride (79 mg, 0.201 mmol), 2-propenoyl chloride (0.016 mL, 0.201 mmol) and A/,/V-diisopropylethylamine (0.070 mL, 0.401 mmol) in W-methyl-2- pyrrolidone (1 mL) was stirred at room temperature for 10 minutes. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (39 mg, 0.095 mmol, 47 % yield). LCMS (Method B): Rt 1.96 minutes; m/z 412 (MH+).
Example 13:
W-{2-(4-acryloyl-1 -piperaziny])-6-[(methy^
6]pyridin-2-amine
Figure imgf000163_0002
A mixture of /V-[6-[(methyloxy)methyl3-2-(1-piperazinyl)-4-pyrimidinyl]t1 ,3]thiazolo[5,4- jb]pyridin-2-amine hydrochloride (49 mg, 0.137 mmol), 2-propenoyl chloride (0.012 mL, 0.137 mmol) and W,A/-diisopropyiethylamine (0.048 mL, 0.274 mmol) in /V-methyl-2- pyrrolidone (1 mL) was stirred at room temperature for 10 minutes. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (24 mg, 0.058 mmol, 42 % yield). LCMS (Method B): Rt 2.17 minutes; m/z 412 (MH+). Example 14:
W-(c/s-2-{[4-[(methyloxy)methyl]-6^
pyrimidiny!3amino}cyclohexyl}-2-propenamide
Figure imgf000164_0001
A mixture of A/2-[c s-2-aminocyclohexyl]-6-[(methyioxy)methyl]- \Ti-[1 ,3]thiazolo[5,4~ /b]pyridin-2-y!-2,4-pyrimidinediamine (70 mg, 0.182 mmol), 2-propenoyi chloride (0.015 mL, 0.182 mmo!) and W.W-diisopropylethylamine (0.063 mL, 0.363 mmol) in N-methyl-2- pyrroSidone (1 mL) was stirred at room temperature for 10 minutes. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (1 1 mg, 0.025 mmoi, 14 % yield). LCMS (Method B): Rt 2.23 minutes; m/z 440 (MH+).
Example 15:
W-(4-{[4-[(methyloxy)methyl]-6-([1,3]thiazolo[5t4-b]pyridin-2-ylami
pyrimidinyl]amino}cyclohexyl)-2-propenamide
Figure imgf000164_0002
A microwave vial was charged with W-[6-[(methy!oxy)methyl]-2-(methylsulfonyl)-4- pyrimidinyl][1.SJthiazolotS^-tojpyridliri^-amine (200 mg, 0.569 mmol), 1 ,4- cyciohexanediamine (97 mg, 0.854 mmol) and isopropanol (4 mL). The vial was sealed and was heated in a Biotage initiator microwave at 150 °C for 30 minutes. After cooling, the reaction was filtered under reduced pressure and the solid was washed with isopropanol. The filtrate was evaporated to dryness to give a yellow solid. The solid was dissolved in W-methyl-2-pyrrolidone (3 mL) and treated with 2-propenoyl chloride (0.096 mL, 1.138 mmol) and /VJV-diisopropylethyiamine (0.596 mL, 3.41 mmol). The reaction was stirred at room temperature for 5 minutes. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) twice to afford the title compound (45 mg, 0.102 mmoi, 18 % yield). LCMS (Method B): Rt 2.08 minutes; m/z 440 (MH+). Example 16:
W-(irans-4-{[4-[(methyloxy)methyl]-6-([1 ,3]thiazolo[5,4- )]pyridin-2-ylamino)-2- pyrimidinyl]amino}cyclohexyl)-2-propenamide
Figure imgf000165_0001
A mixture of W2-(irans-4-aminocyciohexyl)-6-[(methyloxy)methyl]-A/''-[1 ,3]thiazo!o[5,4- b]pyridin-2-yl-2,4-pyrtmidinediamine (50 mg, 0.13 mmol), 2-propenoy! chloride (0.014 mL, 0.169 mmoi) and Λ/,/V-diisopropylethylamine (0.136 mL, 0.778 mmol) in /V-methyl-2- pyrrolidone (2 mL) was stirred at room temperature for 10 minutes. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) twice to afford the title compound (31 mg, 0.071 mmol, 54 % yield). LCMS (Method C): Rt 1.55 minutes; m/z 440 (MH+).
Example 17:
W2-[(3R)-1 -aery loyl-S-pyrrolidinyll-e-KmethyloxyJmethyll-AT'-tl ,3]thiazoIo[5,4- i)lpyndin-2-y!-2,4-pyrimidinediamine
Figure imgf000165_0002
A mixture of 6-[(methyloxy)methyl]-/V2-[(3R)-3-pyrroiidinyl]-A/',-[1 ,3]thiazolo[5,4-b]pyndin-2- yl-2,4-pyrimidinediamine hydrochloride (73 mg, 0.185 mmol), 2-propenoyl chloride (0.017 mL, 0.204 mmol) and /V,W-diisopropylethylamine (0.071 mL, 0.408 mmol) in /V-methyl-2- pyrrolidone (1 mL) was stirred at room temperature for 10 minutes. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) twice to afford the title compound (37 mg, 0.09 mmol, 49 % yield). LCMS (Method B): Rt 1.96 minutes; m/z 412 (MH+).
Example 18:
W-{2-[(1-acryloyl-4-piperidinyl)oxy]-6-[(methyloxy)methyi]-4- pyrimidinyl}|;i ,3]thiazolo[5,4- )]pyridin-2-amine
Figure imgf000166_0001
A mixture of W-[6-[{methyloxy)methyl]-2-(4-piperidinyioxy)-4-pyrimidinyl][1 ,3]thiazolo[5>4- b]pyridin-2-amine hydroch!oride (80 mg, 0.196 mmol), 2-propenoyl chloride (0.017 mL, 0.196 mmol) and W,A/-diisopropy!ethylamine (0.068 mL, 0.391 mmol) in A/-methyl-2- pyrrolidone (1 mL) was stirred at room temperature for 10 minutes. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) twice to afford the title compound (20 mg, 0.047 mmol, 24 % yield). LC S (Method D): Rt 0.78 minutes; m/z 427 ( H+).
Example 19:
(2E)-4-(dimethylamino)-N-(2-{[4-[(me
ylamino)-2-pyrimidinyl]amino}ethyl)-2-butenamide
Figure imgf000166_0002
A mixture of (2£)-4-(dimethylamino)-2-butenoic acid hydrochloride (37.5 mg, 0.226 mmo!), Λ/,/V-diisopropylethylamine (0.079 mL, 0.453 mmo!) and 0-(7-azabenzotriazol-1-y!)- A/JV,A/',A/'-tetramethyluronium hexafiuorophosphate (86 mg, 0,226 mmol) in /V-methyl-2- pyrrolidone (1 mL) was stirred at room temperature for 2 minutes. A solution of Λ/2-(2- aminoethy -e-^methyioxyJmethyll-A^-fl .SjthiazoloiS^-j jpyridin^-yl^^- pyrtmidtnediamine (50 mg, 0.151 mmol) and WJV-diisopropylethylamine (0,079 mL, 0.453 mmol) in W-methyl-2-pyrrolidone (1 mL) was added to the previous solution and the reaction was stirred at room temperature for 1 hour. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (40 mg, 0.09 mmol, 60 % yield). LCMS (Method B): Rt 1.89 minutes; m/z 443 (MH+).
Example 20: W2-{1 -[(2E)-4-(dimethy!amino)-2^ute^
-fe]pyridin-2-yl-2,4-pyrimidinediamine
Figure imgf000167_0001
A mixture of (2£)-4-(dimethylamino)-2-butenoic acid hydrochloride {14.62 mg, 0.088 mmol), W./V-diisopropylethylamine (0.031 mL, 0.177 mmol) and 0-(7-azabenzotriazol-1 - yl)-A/;/V, \/',A/-tetramethyiuronium hexafluorophosphate (33.6 mg, 0.088 mmol) in N- methyl-2-pyrrolidone (0.5mL) was stirred at room temperature for 2 minutes, A solution of 6-[(methyloxy)methyl]-A/2-4-piperidinyl-A/4-t1 ,3]thiazoloE5,4-0]pyridin-2-yl-2,4- pyrimidinediamine hydrochloride (24 mg, 0.059 mmol) and v,/V-diisopropylethylamine (0.031 mL, 0.177 mmol) in A/-methyl-2-pyrrolidone (0.5 mL) was added to the previous solution and the reaction mixture was stirred at room temperature for 1 hour. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (8 mg, 0.016 mmol, 28 % yield). LCMS (Method D): Rt 0.82 minutes; m/z 483 (MH+).
Example 21 :
W^1-[(2E)-4-(dimethylamino)-2-butenoyl]-3^
-f)]pyridin-2-yl-2,4-pynmidinediamine
Figure imgf000167_0002
A microwave via! was charged with A/-[6-[(methyloxy)methyl]-2-(methylsulfonyl)-4- pyrimidinyl][1 ,3]thiazolo[5,4-i)]pyridin-2-amine (500 mg, 1.423 mmol), 1 , 1-ditnethylethyl 3- amino-1 -piperidinecarboxylate (570 mg, 2.85 mmol) and isopropanol (5 mL). The microwave vial was sealed and was heated in a Biotage Initiator microwave at 150 °C for 1.5 hours. The residue was purified by chromatography on silica using a gradient elution from 0 to 15 % methanol (+1 % triethylamine) in dichloromethane. After evaporation of the desired fractions to dryness, the residue was dissolved in dichloromethane and treated with 4M HCI in dioxane (0.6 mL, 2,4 mmol) at room temperature. The solution was stirred at room temperature overnight. The reaction mixture was filtered under reduced pressure, dried to give 185 mg of an intermediate so!id. A mixture of (2£)-4-(dimethylamino)-2- butenoic acid hydrochloride (30.5 mg, 0.184 mmol), /V,/V-diisopropylethylamine (0.064 mL, 0.368 mmo!) and 0-(7-azabenzotriazol-1 -yl)-W,/\/, \/', \/-tetramethyluronium hexafluorophosphate (69.9 mg, 0.184 mmol) in A/-methyl-2-pyrrolidone (1 mL) was stirred at room temperature for 2 minutes. A solution of the intermediate solid (50 mg) and N,N- ditsopropylethylamine (0.064 mL, 0.368 mmol) in A/-methyl-2-pyrrolidone (1 mL) was added to the previous solution and the reaction mixture was stirred at room temperature for 1 hour. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (22 mg, 0.046 mmol, 3 % yield). LCMS (Method D): Rt 0.81 minutes; m/z 483 ( H+). Example 22:
N-(2-{[4-(4-morpholinylmethyl)-6-(^
pyrimidinyl]amino}ethyl)-2-propenamide
Figure imgf000168_0001
A mixture of /\/2-(2-aminoethyl)-6-(4-morpholinylmethyi)-A/'i-[1 ,3]thiazolo[5,4-i)]pyridin-2-yl- 2,4-pyrimidinediamine (31 mg, 0.08 mmol), Λ/,/V-diisopropylethylamine (0.028 mL, 0.160 mmol) and 2-propenoyl chloride (0.009 mL, 0.112 mmol) in W-methyl-2-pyrro!idone (0.8 mL) was stirred at room temperature for 30 minutes. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (22 mg, 0.05 mmol, 62 % yield). LCMS (Method B): Rt 1.84 minutes; m/z 441 (MH+).
Example 23:
W-(frans-2-{[4-(4-morpholinylmethyI)-6-([1 ,3]thiazolo[5,4- 3]pyridin-2-ylamino)-2- pyrimidinyl]amino}cyclohexyl)-2-propenamide
Figure imgf000169_0001
A mixture of W2-[fra/?s-2-aminocyclohexyl]-6-(4-morpholinylmethyi)-/V4-[1 ,3]th i>]pyridtn-2-yl-2,4-pyrimidinediamine (45 mg, 0.102 mmol), W,A/-diisopropylethylamine (0.021 mL, 0.123 mmoi) and 2-propenoyl chloride (0.009 mL, 0.1 12 mmol) in W-methy!-2- pyrrolidone (0.8 mL) was stirred at room temperature for 30 minutes. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (20 mg, 0.04 mmol, 40 % yield). LCMS (Method B): Rt 2.22 minutes; m/z 495 (MH+). Example 24:
W2-(1 -aciyloyl-3-piperidinyi)-6-(4-morphoIiny^
yl-2,4-pyrimidinediamine
Figure imgf000169_0002
A mixture of 6-(4-morpholinylmethyi)-/V2-3-piperidinyl-A/'i-[1 ,3]thiazolo[5,4-£)]pyndin-2~yl- 2,4-pyrimidinediamine dihydrochloride (70 mg, 0.14 mmol), N,A/-diisopropylethylamine (0.073 mL, 0.42 mmol) and 2-propenoyl chloride (0.023 mL, 0.28 mmol) in W-methyl-2- pyrroiidone (0.8 mL) was stirred at room temperature for 30 minutes. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (20 mg, 0.042 mmo!, 30 % yield). LCMS (Method B): Rt 2.01 minutes; m/z 481 (MH+).
Example 25:
W-(ira/?s-4-{[4-(4-morpholinylmethyI)-6-([1 ,3]thiazolo[5,4- ?3pyridin-2-ylamino)-2- pyrimidinyl]amino}cyclohexyl)-2-propenamide
Figure imgf000170_0001
A mixture of ^-{/rans-^aminocyclohexyiJ-e-t^morpholinylmethylJ-A^-tl ^lthiazolotS^- /b]pyridin-2-yl-2,4-pynmid!nediamine (54 mg, 0.123 mmol), 2-propenoyl chloride (0.020 mL, 0.245 mmol) and fyW-diisopropylethylamine (0.043 mL, 0.245 mmol) in A/-methyl-2- pyrrolidone (1 mL) was stirred at room temperature for 10 minutes. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (27 mg, 0.055 mmol, 44 % yieid). LCMS (Method B): Rt 2.06 minutes; m/z 495 (MH+). Example 26:
W2-[(3R)-1-acryloyl-3-pyrrolidinylH^
b]pyridin-2-yl-2,4-pyrimidinediamine
Figure imgf000170_0002
A mixture of 6-(4-morpholinyimethy!)-W2-[(3/:?)-3-pyrrolidinyl3-A/',-[1 ,3]thiazolo[5,4-jb]pyridin- 2-yl-2,4-pyrimidinediamine dihydrochloride (48 mg, 0.099 mmol), 2-propenoyl chloride (0.016 mL, 0.198 mmol) and A/JV-diisopropylethylarnine (0.052 mL, 0.297 mmol) in N- methyl-2-pyrro!idone (1 mL) was stirred at room temperature for 10 minutes. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (16 mg, 0.034 mmol, 35 % yield). LCMS (Method D): Rt 0.75 minutes; m/z 467 (MH+).
Example 27:
W-(c s-4-{[4-(4-morpholinylmethyl)-6-([1 ,3]thiazolo[5,4-J ]pyridin-2-ylamino)-2- pyrimidinyl3amino}cyclohexyl)-2-propenamide
Figure imgf000171_0001
A mixture of ^-(c s^-aminocyciohe y -e-f^morpholinylmethy -W^-tl .SIthiazoloiS^- ]pyridin-2-yl-2,4-pyrimidinediamine dihydrochioride (70 mg, 0.136 mmol), 2-propenoy! chloride (0.022 mL, 0.273 mmol) and A/,A/-diisopropylethylamine (0.071 mL, 0.409 mmol) in A/-methyl-2-pyrrolidone (1 mL) was stirred at room temperature for 10 minutes. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (17 mg, 0.034 mmol, 25 % yield). LCMS (Method B): Rt 2.03 minutes; m/z 495 (MH+). Example 28:
Wz-(1-acryloyi-4-piperidinyl)-6-(4-morpholinylm
yl-2,4-pyrimidinediamine
Figure imgf000171_0002
A mixture of 6-(4-morpho!inylmethyl)-W2-4-piperidinyl-W^-[1 ,3]thiazo!o[5,4-0]pyridin-2-yl- 2,4-pyrimidinediamine dihydrochioride (70 mg, 0.14 mmol), /V,A/-diisopropylethylamine (0.147 mL, 0.841 mmol) and 2-propenoyl chloride (0.028 mL, 0.35 mmol) in W-methyl-2- pyrrolidone (0.6 mL) was stirred at room temperature for 30 minutes. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the titie compound (18 mg, 0.037 mmol, 27 % yield). LCMS (Method B): Rt 1.98 minutes; m/z 481 (MH+).
Example 29:
W2-[(3S)-1 -acry!oyl-3-pyrrolidinyl]-6-(4-morpholinylmethyl)-W4-[1 ,3]thiazoloE5f4- i)]pyridin-2-yl-2,4-pyrimidinediamine
Figure imgf000172_0001
A mixture of 6-(4-morpholinylmethyl)-/V2-[(3S)-3-pyrroiidiny^
2-yi-2,4-pyrimidinediamine dihydrochloride (50 mg, 0.103 mmol), 2-propenoyl chloride (0.017 mL, 0.206 mmoi) and W,W-diisopropylethylamine (0.054 mL, 0.309 mmo!) in N- methyi-2-pyrrolidone (1 mL) was stirred at room temperature for 10 minutes. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (13 mg, 0.028 mmol, 27 % yield). LCMS (Method D): Rt 0.75 minutes; m/z 467 (MH+). Example 30:
W-(c/'s-2-{[4-(4-morpholinylmethyl)-6-([1,3]thiazolo[5,4-/j]pyridin-2-ylam
pyrimidinyl]amino}cyclohexyl)-2-propenamide
Figure imgf000172_0002
A mixture of /V2-Ec/s-2-aminocyciohexyl]-6-(4-morpholiny!methy[)-W'i-[1 ,3]thiazolo[5,4- /b]pyridin-2-yl-2,4-pyrimidinediamine (51 mg, 0.1 16 mmol), A ,A/-diisopropylethyiamine (0.024 mL, 0.139 mmoi) and 2-propenoyl chloride (0.01 mL, 0.127 mmol) in /V-methyl-2- pyrrolidone (0.8 mL) was stirred at room temperature for 30 minutes. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (23 mg, 0.046 mmoi, 40 % yield). LCMS (Method B): Rt 2.17 minutes; m/z 495 (MH+).
Example 31 :
W-(1 -{6-[(6-faromo-1 )3-benzothiazol-2-yl)amino]-4-[(2-methyi-1 H-imidazol-1 - yl)methyl3-2-pyridinyl}-3-piperidinyi)-2-propenamide
Figure imgf000173_0001
A mixture of A/-{6-(3-amino-1-piperidinyl)-4-[(2-methyl-1 H-imidazo!-1-yl)methyl]-2- pyridinyi}-6-bromo-1 ,3-benzothiazol-2-amine (50 mg, 0.1 mmol), 2-propenoyl chloride (0.016 mL, 0.201 mmol), N,N-diisopropyiethylamine (0.053 mL, 0.301 mmol) and N- methyi-2-pyrrolidone (0.8 mL) was stirred at room temperature for 2 hours. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (15 mg, 0.027 mmol, 27 % yield). LCMS (Method B): Rt 2.76 minutes; m/z 552, 554 (MH+). Example 32:
W-[2-(4-acryloyl-1 -piperazinyl)-6-(4-morpholinylmethyl)-4- pyrimidinyl][1 ,3]thiazolo[5,4-Jt)]pyridin-2-amine
Figure imgf000173_0002
A mixture of A/-[6-(4-morphoiinylmethyi)-2-(1-piperazinyl)-4-pyrimidinyl][1 ,3]thiazolo[5,4- jb]pyridin-2-amine (50 mg, 0.121 mmol), 2-propenoyl chloride (0.02 mL, 0.242 mmol) and A/,A/-diisopropylethylamine (0.042 mL, 0.242 mmol) in A/-methyl-2-pyrro!idone (1 mL) were stirred at room temperature for 10 minutes. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (15 mg, 0.032 mmoi, 27 % yield). LCMS (Method B): Rt 2.10 minutes; m/z 467 (MH+).
Example 33:
W-[2-[(1 -acryloyl-4-piperidinyl)oxy]-6-(4-morphoiiny!methyl)-4- pyrimidinyi3[1 ,3]ihiazolo[5,4-Jb]pyridin-2-amine
Figure imgf000174_0001
A mixture of /V-[6-(4-morpho!inylmethyl)-2-(4-piperidinyloxy)-4- pyrimidinyl][1 ,3]thiazoio[5,4-0]pyridin-2-am!ne dihydrochloride (50 mg, 0.1 nrtmol), 2- propenoyl chloride (0.016 mL, 0.2 mmoi) and Λ/,/V-diisopropylethylamine (0.052 mL, 0.3 mmoi) in A/-methyl-2-pyrrolidone (1 mL) were stirred at room temperature for 10 minutes. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (15 mg, 0.031 mmoi, 31 % yie!d). LCMS (Method B): Rt 1.91 minutes; m/z 482 (MH+). Example 34:
W-(c/'s-3-{[4-(4-morpholinylm ^
pyrimidinyl]amino}cyclohexyl)-2-propenamide
Figure imgf000174_0002
A mixture of A/ -(3-aminocyclohexyl)-6-(4-morpholinylmethyl)-A/4-t1 ,3]thiazolo[5,4- d]pyridin-2-yl-2,4-pynmidinediamine (94 mg, 0.213 mmoi), Λ/,/V-diisopropylethylamine (0.1 12 mL, 0.640 mmoi) and 2-propenoyl chloride (0.034 mL, 0.427 mmoi) in W-methyl-2- pyrroiidone (0.8 mL) was stirred at room temperature for 1.5 hours. The mixture was subjected twice to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (21 mg, 0.042 mmoi, 20 % yield). LCMS (Method B): Rt 2.05 minutes; m/z 495 (MH+).
Example 35:
W-(frans-3-{[4-(4-morpholinylmethyl)^
pyrimidinyl]amino}cyclohexyl)-2-propenamide
Figure imgf000175_0001
A mixture of W2-(3-aminocyc!ohexyl)-6-(4-morphoiinylmethyl)-A/I'-[1 ,3]thiazolo[5l4- 0]pyridin-2-yl-2,4-pyrimidinediamine (94 mg, 0.213 mmol), A/,A/-diisopropylethylamine (0.112 mL, 0.64 mmol) and 2-propenoyi chloride (0.034 mL, 0.427 mmol) in W-methyl-2- pyrrolidone (0.8 mL) was stirred at room temperature for 1.5 hours. The mixture was subjected twice to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (10 mg, 0.02 mmol, 10 % yield). LCMS (Method B): Rt 2.1 1 minutes; m/z 495 (MH+). Example 36:
W -[1-(chloroacetyl)-3^iperidmyl]-6-(4-mor^
-pyrimidinediamine
Figure imgf000175_0002
Chloroacetyl chloride (0.008 mL, 0.1 mmoi) was added to a cooled (0 °C, ice bath) solution of 6-(4-morpho!inyimethyl)-A/5-3-piperidiny!- \/'i-[1 ,3]thiazolo[5,4-0]pyridin-2-yl-2,4- pyrimidinediamine dihydrochloride (50 mg, 0.1 mmol) and A/JV-diisopropylethylamine (0.07 mL, 0.4 mmoi) in A/-methy!-2-pyrroIidone (0.8 mL). The reaction was stirred at 0 °C for 2 hours. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (13 mg, 0.026 mmol, 26 % yield). LCMS (Method B): Rt 2.10 minutes; m/z 503 (MH+).
Example 37:
/V-[2-(2-acryloyltetrahydro-1 (2H)-pyridazinyl)-6-(4-morpholinylmethyl)-4- pyrimidinyl][1 ,3]thiazolo[5,4-i ]pyridin-2-amine
Figure imgf000176_0001
A microwave vial was charged with W-[2-chloro-6-(4-morpholinylmethyl)-4- pyrimidinyl][1 ,3]thiazolo[5,4-b]pyridin-2-amine (100 mg, 0,276 mmol), hexahydropyridazine hydrochloride (132 mg, 0.827 mmol) and Λ/,/V-diisopropylethylamine (0.144 mL, 0.827 mmol). The microwave via! was sealed and heated in the Biotage Initiator microwave system at 170 °C for 45 minutes. The solvent was evaporated to dryness. The brown residue was dissolved in A/-methyl-2-pyrrolidone (2 mL), N,N- diisopropylethylamine (0.289 mL, 1.654 mmol) and 2-propenoyl chloride (0.134 mL, 1.654 mmol) were added and the reaction was stirred at room temperature for 1 hour. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (20 mg, 0,043 mmol, 16 % yield). LCMS (Method B): Rt 2.22 minutes; m/z 467 (MH+).
Example 38:
N2-(1 -{[(4-fluorophenyl)oxy]acetyl}-3-piperidinyl)-6-(4-morpholinylmethyl)-W4- [1 ,3]thiazolo[5,4-i)]pyridin-2-yl»2,4-pyrimidinediamine
Figure imgf000176_0002
A mixture of 4-fluorophenoxyacetic acid (25.5 mg, 0, 15 mmol), W,A/-diisopropylethylamine (0.052 mL, 0.3 mmol) and 0-(7-azabenzotnazol-1 -yl)- V, )WJ/V' /V'-tetramethy!uronium hexafluorophosphate (57.1 mg, 0.15 mmol) in W-methyl-2-pyrrolidone (1 mL) was stirred at room temperature for 2 minutes. A solution of 6-(4-morpholinylmethyl)-W2-3-piperidinyi-A/i'- [1 ,3]thiazolo[5,4-i)]pyridin-2-yl-2,4-pyrimidinediamine dihydroch!oride (50 mg, 0.1 mmol) and W,A/-diisopropylethylamine (0.052 mL, 0.3 mmol) in A/-methyl-2-pyrrolidone (1 mL) was added to the previous solution and the reaction was stirred at room temperature for 1 hour. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (32 mg, 0,055 mmol, 55 % yield). LCMS (Method B): Rt 2.37 minutes; m/z 579 (MH+).
Example 39:
2-chloro-W-(2-{[4-(4-morpholinylmethyl)-6-([1 ,33thiazolo[5,4-Jb}pyridin-2-ylami
ethyl)acetamide
Figure imgf000177_0001
Chloroacetyl chloride (0,013 mL, 0.163 mmol) was added to a cooled (0 °C, ice bath) solution of /V2-(2-aminoethyl)-6-(4-morphollnylmethyl)-A ''-[1 ,3]thiazolo[5,4-jb]pyridin-2-yl- 2,4-pyrimidinediamine dihydrochloride (75 mg, 0.163 mmoi) and N,N~ diisopropylethylamine (0.1 14 mL, 0.653 mmol) in A/-methyl-2-pyrrolidone (1 mL). The reaction was stirred at 0 °C for 2 hours. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (25 mg, 0.054 mmol, 33 % yield). LCMS (Method B): Rt 1.89 minutes; m/z 463 (MH+).
Example 40:
{2£)-4,4)4-trifluoro-W-(2-{[4-(4-morpholinylmethyl)-6-([1 ,3]thiazolo[5,4-Jb3pyridin-2- ylamino)-2-pyrimidinyl]amino}ethyl)-2-butenamide
Figure imgf000177_0002
A mixture of (2E)-4,4,4-trifluoro-2-butenoic acid (34.3 mg, 0.245 mmol), N,N- diisopropylethylamine (0.086 mL, 0.49 mmoi) and 0-(7-azabenzotriazol-1 -yl)-W,N,/V' A/- tetramethyiuronium hexafiuorophosphate (93 mg, 0.245 mmoi) in A/-methyi-2-pyrrolidone (1 mL) was stirred at room temperature for 2 minutes. A solution of A/2-(2-aminoethyl)-6- (4-morphoiinyimethyl)-A/'i-[1 ,3]thiazolo[5,4-jb]pyridin-2-yl-2l4-pyrimidinediamine
dihydrochloride (75 mg, 0.163 mmol) and Λ/JV-diisopropy!ethy!amine (0,086 mL, 0.49 mmol) in A/-methy!-2-pyrrolidone (1 ml_) was added to the previous solution and the reaction was stirred at room temperature for 16 hours. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (32 mg, 0.063 mmol, 39 % yield). LCMS (Method B); Rt 2.25 minutes; m/z 509 (MH+).
Example 41 :
2-[(4-fluorophenyl)oxy]-W-(2-{[4-(4-mor^
2-ylamino)-2-pyrimidinyl]amino}ethy[)acetamide
Figure imgf000178_0001
A mixture of 4-fluorophenoxyacetic acid (41.7 mg, 0.245 mmol), N,N- diisopropylethylamine (0.086 mL, 0.49 mmol) and 0-(7-azabenzotriazol-1-yl)-WJV,A/' W- tetramethyluronium hexafiuorophosphate (93 mg, 0.245 mmol) in /V-methyi-2-pyrro!idone (1 mL) was stirred at room temperature for 2 minutes. A solution of A/2-(2-aminoethyl)-6- (4-morpholinylmethyi)-A/4-[1 ,3]thiazolo[5,4-jb]pyridin-2-yl-2,4-pyrimidinediamine
dihydrochloride (75 mg, 0.163 mmol) and W,W-diisopropylethylamine (0.086 mL, 0.49 mmol) in A/-methyl-2-pyrro!idone (1 mL) was added to the previous solution and the reaction was stirred at room temperature for 16 hours. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (42 mg, 0.078 mmol, 48 % yield). LCMS (Method B): Rt 2.23 minutes; m/z 539 (MH+).
Example 42:
/V_{1 -[4-(4-morpholinylmethyl)-6-([1 ,3]thiazolo[5,4- i]pyridin-2-ylamino)-2-pyridinyIj- -piperidinyl}-2-propenamide
A mixture of A/-[6-(3-amino-1-piperidinyl)-4-(4-morpholinylmethyl)-2- pyridinyl][1 ,33thiazolo[5,4-£)]pyridin-2-amine (29 mg, 0.068 mmol), 2-propenoyl chloride (0.007 mL, 0.082 mmo!) and A/,A/-diisopropylethylamine (0.024 mL, 0.136 mmol) in N- methyl-2-pyrrolidone (1 mL) were stirred at room temperature for 10 minutes. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (28 mg, 0.058 mmol, 86 % yield). LCMS (Method B): Rt 2.20 minutes; m/z 480 (MH+).
Example 43:
W-(c s-3-{[4-(4-morphoMnylmethyl)-6-([1 ,3]thiazolo[5,4-fc]pyridin-2-yl
pyridinyl]amino}cyclohexyl)-2-propenamide
Figure imgf000179_0001
A mixture of
Figure imgf000179_0002
i>]pyridin-2-yl-2,6-pyridinediamine (20 mg, 0.045 mmol), 2-propenoyl chloride (0.004 mL, 0.055 mmol) and A/,W-diisopropylethylamine (0.016 mL, 0.091 mmol) in W-methyl-2- pyrro!idone (1 mL) was stirred at room temperature for 10 minutes. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (14 mg, 0.028 mmol, 63 % yield). LCMS (Method B): Rt 2.18 minutes; m/z 494 (MH+).
Example 44:
/V-(fra/7S-3-{[4-{4-morpholinylmethyl)-6-([1 ,3]thiazoio[5,4-b]pyridin-2-ylam
pyridinyl]amino}cyclohexyl)-2-propenamide
Figure imgf000179_0003
A mixture of W-Er ans-S-aminocyclohexylj-^t^morpholinylmethy -W-Il ^jthiazoloIS^- i>]pyridin-2-yl-2,6-pyridinediamine (10 mg, 0.023 mmol), 2-propenoyl chloride (0.002 mL, 0.027 mmol) and A/./V-diisopropylethylamine (0.008 mL, 0.045 mmol) in /V-methyi-2- pyrrolidone (1 mL) was stirred at room temperature for 10 minutes. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the titje compound (4 mg, 0.008 mmol, 35 % yield). LC S (Method B): Rt 2.25 minutes; m/z 494 (MH+).
Example 45:
W-(2-{[4-(4-morpholinylmethyl)-6-([1 ,3]thiazolo[5,4-J 3pyridin-2-ylamino)-2- pyridrnyl]amino}ethyI)-2-propenamide
Figure imgf000180_0001
A microwave vial was charged with 1 , 1-dimethylethyl (2-aminoethyl)carbamate (443 mg, 2.76 mmol), A/-[6-chloro-4-(4-morpholinylmethyl)-2-pyridinyl][1 ,3]thiazolo[5,4-ib]pyridin-2- amine (100 mg, 0.276 mmol) and ethylene glycol (1 mi). The vial was sealed and was heated in a Biotage initiator microwave to 200 °C for 1 hour. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford 35 mg of the amine intermediate. The intermediate (32 mg), 2- propenoyl chioride (0.008 mL, 0.1 mmol) and Λ , V-diisopropylethylamine (0.029 mL, 0.166 mmol) in A/-methyl-2-pyrrolidone (1 mL) was stirred at room temperature for 10 minutes. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (2.8 mg, 0.006 mmoi, 2 % yield). LCMS (Method B): Rt 1.96 minutes; m/z 440 (MH+). Example 46:
W-c s-2-{[4-(4-morpholinylmethyl)-6-{[1,3]thiazolo[5t4-ij]pyridin-2-ylami
pyridinyl]amino}cyclohexyl)-2-propenamide
Figure imgf000181_0001
A mixture of W-[c/s-2-aminocyclohexyl]-4-(4-morpholinylmethyl)-W'-[1 ,3]thiazolo[5,4- b]pyridin-2-yl-2,6-pyridinediarriine (18 mg, 0.041 mmol), 2-propenoy! chloride (0.005 mL, 0.049 mmoi) and A/,A/-diisopropyiethyiamine (0.014 mL, 0.082 mmol) in /V-methyi-2- pyrrolidone (1 mL) was stirred at room temperature for 10 minutes. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (3.8 mg, 0.008 mmol, 19 % yield). LC S (Method B): Rt 2.29 minutes; m/z 494 (MH+).
Example 47:
W-ii ans^-i^-t^morpholinylmethy -e-ifl^lthiazoloiSAftlpyridin
pyridinyl]amino}cyclohexyl)-2-propenamide
A mixture of A/-[fra 7s-2-aminocyclohexyl]-4-(4-morpholinylmethyl)-A/'-[1 ,3]thiazolo[5>4- 63pyridin-2-yl-2,6-pyridinediamine (20 mg, 0,045 mmol), 2-propenoyl chloride (0.004 mL, 0.055 mmol) and Λ/,/V-diisopropylethylamine (0.016 mL, 0.091 mmol) in /V-methyl-2- pyrro!idone (1 mL) was stirred at room temperature for 10 minutes. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (15.6 mg, 0.032 mmol, 70 % yield). LCMS (Method B): Rt 2.34 minutes; m/z 494 (MH+),
Example 48:
6-(4-morphoiinylmethyl)-W4-[1 ,3]thiazolo[5,4-b]pyridin-2-yUW l-[(2£)-4,4,4-tn 2-butenoyl]-3-piperidinyl}-2,4-pyrimidinediamine
Figure imgf000182_0001
A mixture of (2£)-4,4,4-trifluoro-2-butenoic acid (29.4 mg, 0.21 mmol), N,N- diisopropylethylamine (0.073 mL, 0.42 mmol) and 0-(7-azabenzotriazol-1-yl)-W,A/,W' A/'- tetramethyluronium hexaf!uorophosphate (80 mg, 0.21 mmol) in A/-methyl-2-pyrroiidone (1 mL) was stirred at room temperature for 2 minutes. A solution of 6-(4- morpholinylmethy -W^S-piperidinyl-A/^-Il ^jthiazolo^^-^pyridin^-yl^^- pyrimidinediamine dihydrochloride (70 mg, 0.14 mmol) and A/,A/-diisopropylethylamine (0.073 mL, 0.42 mmol) in A/-methyl-2-pyrrolidone (1 mL) was added to the previous solution and the reaction was stirred at room temperature for 2 hours. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier), followed by a second purification by mass directed automated preparative HPLC (formic acid modifier) to afford the title compound (8 mg, 0.015 mmol, 10 % yield). LCMS (Method B): Rt 2.33 minutes; m/z 549 (MH+). Example 49:
W-methyl-W-{2-{i4-(4-morpholiny!methyl)-6-([1 ,3]thiazolo[5,4-b]pyridin-2-ylam pyrimidinyl]amino}ethyl)-2-propenamide
Figure imgf000182_0002
A mixture of A 2-[2-(methylamino)ethyl]-6-(4-morpholiny!methyi)-A i-[1 ,3]thiazolo[5,4- b]pyridin-2-yl-2,4-pyrimidinediamine dihydrochloride (50 mg, 0.106 mmol), N,N- diisopropy!ethylamine (0.065 mL, 0.372 mmol) and 2-propenoyi chloride (0.015 mL, 0.186 mmol) in A -methy!-2-pyrro!idone (1.5 mL) was stirred at room temperature for 30 minutes. The mixture was subjected twice to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (18 mg, 0.04 mmol, 37 % yield). LCMS (Method B): Rt 2.04 minutes; m/z 455 (MH+).
Example 50:
-{3-[4-(4^θΓρΜοΙϊηνΐΕΤΐθΙΗγΙ)-6-{(;ΐ,3]^Ϊ3ΖθΙο[5,4-ΐ)]ρνπζ1ϊη-2-γΐ3ηιΐηο)-2- pyrimidinyl]phenyl}-2-propenamide
Figure imgf000183_0001
4M HCI in dioxane (2 mL, 8 mmol) was added to a solution of 1 ,1 -dimethyiethyl {3-[4-(4- morpholinylmethy -e-itl ^jthiazo!oIS^-^ yridin^-ylamino)^- pyrimidiny!]phenyl}carbamate (20 mg, 0.038 mmo!) in dich!oromethane (2 mL) and methanol (1 mL). The reaction mixture was stirred at room temperature for 3 hours. The solvent was evaporated to dryness. A mixture of the residue (23 mg), 2-propenoyl chloride (0.005 mL, 0.066 mmol) and A/,A/-dnsopropylethylamine (0.019ml, 0.109 mmol) in N- methyi-2-pyrrolidone (1 mL) was stirred at room temperature for 10 minutes. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (9 mg, 0.019 mmol, 50 % yield). LCMS (Method B): Rt 2.35 minutes; m/z 474 (MH+).
Example 51 :
W 4-(4-morpholinylmethyl)-6-([1 ,3]thiazolo[5,4- )]pyridin-2-ylamino)-2-pyridin propenamide
Figure imgf000183_0002
A round bottom flask was charged with 4-(4-morpholinylmethyl)- V-(1 ,3]thiazoio[5,4- b]pyridin-2-yl-2,6-pyridinediamine (20 mg, 0.058 mmol) in anhydrous A/-methyl-2- pyrrolidone (1 mL). 2-Propenoyl chloride (0.007 mL, 0.088 mmol) was added and the reaction was stirred at room temperature under nitrogen atmosphere for 16 hours. A further portion of 2-propenoyl chloride (0.05 mL, 58.83 mmoi) was added and the reaction stirred at room temperature for 4 hours under nitrogen atmosphere. Triethyiamine (0.1 mL, 0.72 mmoi) was added. The reaction was stirred at room temperature for 1 hour under nitrogen atmosphere. The reaction mixture was partitioned between dichloromethane (20 mL) and saturated aqueous sodium carbonate (20 mL). After separation, the organic extract was dried using a hydrophobic frit and evaporated to dryness. The product was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (2.3 mg, 0.0058 mmol, 10 % yield). LCMS (Method D): Rt 0.86 minutes; m/z 397 (MH+).
Example 52:
W44-(4-morpholinylmethyl)-6-([1,3]thiazolo[5,4-ib]pyNdin-2-ylamino)-2(1 H)- pyrimidinylidene]-2-propenohydrazide
Figure imgf000184_0001
A microwave via! was charged with A/-[2-chloro-6-(4~morpholinylmethyl)-4- pyrimidinyl][1 ,3]thiazolo[5,4-0]pyridin-2-amine (100 mg, 0.276 mmol), rert-butyl carbazate (109 mg, 0.827 mmol) and isopropanol (3 mL). The microwave vial was sealed and heated in a Biotage Initiator microwave at 170 °C for 45 minutes. A second portion of tert- butyl carbazate (109 mg, 0.827 mmol) was added, the vial was sealed and heated in a Biotage Initiator microwave at 170 °C for 30 minutes. The mixture was evaporated to dryness. The yellow solid was suspended in dichloromethane (5 mL) and treated with 4M HCI in dioxane (1 mL, 4 mmol). The reaction was stirred at room temperature for 16 hours. The mixture was evaporated to dryness. The yellow solid was dissolved in N- methyl-2-pyrrolidone (2 mL) and treated with /V,/V-diisopropylethylamine (0.241 mL, 1.378 mmol) and 2-propenoyl chloride (0.045 mL, 0.551 mmol). The reaction was stirred at room temperature for 2 hours. Further portions of A/,A/-di!Sopropylethylamine (0.5 mL) and 2- propenoyl chloride (0.1 mL) were added and the reaction stirred at room temperature for 2 hours. The product was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (30 mg, 0.073 mmol, 26 % yield). LCMS (Method D): Rt 0.63 minutes; m/z 413 (MH+). Example 53:
W2-[(3S)-1-acryloyl-3-pyrrolidinyl]-^-(6-bromo-1 ,3-benzothiazoi-2-yl)-2,4- pyrimidinediamine
Figure imgf000185_0001
A round bottom flask was charged with 1 ,1 -dimethy!ethyl (3S)-3-({4-[(6-bromo-1 ,3- benzothiazoi-2-yi)amino]-2-pyrimidinyi}amino)-1-pyrrolidinecarboxyiate (50 mg, 0.102 mmol), 4M HCI in dioxane (1 mL, 4.00 mmol), dichloromethane (3 mL) and methanol (2 mL). The reaction was stirred at room temperature for 1 hour. The solvent was evaporated to dryness. The residue was dissolved in A/-methyl-2-pyrrolidone (1 mL) and treated with JV-diisopropylethylamine (0.089 mL, 0.509 mmo!) and 2-propenoyl chloride (0.016 mL, 0.203 mmol). The reaction was stirred at room temperature for 1 hour. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (19 mg, 0.043 mmo!, 42 % yield). LCMS (Method D): Rt 0.98 minutes; m/z 445/447 ( H+).
Example 54:
W-ia-i^-Ke-bromo-l^-benzothiazol^-y aminol-e^-morpholin lmethyi)^- pyrimidinyl]amino}ethyl)-2-propenamide
Figure imgf000185_0002
A round bottom flask was charged with 1 ,1 -dimethylethy! (2-{[4-[(6-bromo-1 ,3- benzothiazol-2-yl)amino]-6-(4-morpholinylmethyl)-2-pyrimidinyl]amino}ethyl)carbamate (50 mg, 0.089 mmol), 4M HCI in dioxane (0.870 mL, 3.48 mmol), dichloromethane (3 mL) and methanol (2 mL). The reaction was stirred at room temperature for 1 hour. The solvent was evaporated to dryness. The residue was dissolved in A/-methyl-2-pyrroiidone (3 mL) and treated with Λ/,/ -diisopropylethylamine (0.077 mL, 0.443 mmoi) and 2- propenoyl chloride (0.014 mL, 0.177 mmol). The reaction was stirred at room temperature for 1 hour. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (26 0.05 mmol, 57 % yield). LCMS (Method D): Rt 0.96 minutes; m/z 518 / 520 (MH+).
Example 55:
^^[(SSi-l -acr loyl-S- yrrolidin ll-Af'-ie-bromo-I .S-benzothiazoi^-yiJ-e-t^
morphoiinylmethyl)-2,4-pyrimidinediamine
Figure imgf000186_0001
A round bottom flask was charged with 1 ,1 -dimethylethyl (3S)-3-{[4-[(6-bromo-1 ,3- benzothiazol-2-yl)amino]-6-(4-morpholinylmethyl)-2-pyrim!dinyl]amino}-1 - pyrrolidinecarboxylate (50 mg, 0.085 mmol), 4M HCI in dioxane (0.832 mL, 3.33 mmol), dichloromethane (3 mL) and methanol (2 mL). The reaction was stirred at room temperature for 1 hour. The reaction mixture was evaporated to dryness. The residue was dissolved in W-methyi-2-pyrrolidone (1 mL) and treated with W,W-diisopropyiethylarnine (0.074 mL, 0.423 mmol) and 2-propenoyl chloride (0.014 mL, 0.169 mmol). The reaction was stirred at room temperature for 1 hour. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (31 mg, 0.057 mmol, 67 % yield) as an off-white solid. LCMS (Method D): Rt 0.99 minutes; m/z 544 / 546 (MH+). Example 56:
W-(1 -actyloyl-4-piperidinyl)-4-(4-morpholiny!methyl)-W'-[1 ,3]thiazolo[5,4-fa]pyridi -2,6-pyridinedlamine
Figure imgf000186_0002
A round bottom flask was charged with 1 , 1-dimethy!ethyl 4-{[4-(4-morpholinylmethyl)-6- ([1 ,3]thiazolo[5,4-i)]pyridin-2-ylamino)-2-pyridinyl]amino}-1-piperidinecarboxylate (30 mg, 0.057 mmol), 4M HCI in dioxane (0.3 mL, 1.2 mmol) and dichloromethane (3 mL). The reaction was stirred at room temperature for 2 hours. The solvent was evaporated to dryness. The solid residue was dissolved in A/-methyl-2-pyrrolidone (1 mL). N,N- diisopropylethy!amine (0.05 mL, 0.285 mmol) was added followed by 2-propenoyi chloride (0.014 mL, 0.171 mmol). The reaction was stirred at room temperature for 1 hour. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (14 mg, 0.029 mmol, 51 % yield) as a brown solid. LCMS (Method D): Rt 0.83 minutes; m/z 480 (MH+).
Example 57:
W-[(3S)-1 -acryloyl-3-pyrrolidinyl]-4-(4-morpholinylmethyl)-W*-[1,3]thiazo
i>]pyridin-2-yl-2,6-pyridinediamine
Figure imgf000187_0001
A round bottom flask was charged with 1 ,1 -dimethylethyi (3S)-3-{[4-(4-morpholinylmethyl)- 6-([1 ,3]thiazo!o[5,4-0]pyridin-2-ylamino)-2-pyridinyl3amino}-1-pyrrolidinecarboxylate (35 mg, 0.068 mmol), 4 HCi in dioxane (0.3 ml, 1.200 mmol) and dichloromethane (3 mL). The reaction was stirred at room temperature for 1 hour. The reaction mixture was evaporated to dryness. The residue was dissolved in A -methyl-2-pyrrolidone (1 mL). N,N- diisopropylethylamine (0.036 ml, 0.205 mmol) and 2-propenoyi chloride (0.011 ml, 0.137 mmol) were added and the reaction was stirred at room temperature for 1 hour. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (18 mg, 0.039 mmol, 57 % yield). LCMS (Method D): Rt 0.80 minutes; m/z 466 (MH+).
Example 58:
W-(1 -acryloyl-3-piperidinyl)-4-(4-morpholinylm
yl-2,6-pyridinediamine, formic acid salt
Figure imgf000188_0001
A round bottom flask was charged with 4-(4-morpho!inylmethyi)-iV-3-pipendinyl-A/'- [1 ,3]thiazolo[5,4-b]pyridin-2-yl-2,6-pyridinediamine (55 mg, 0.129 mmol) and N,N- diisopropylethylamine (0.068 mL, 0.388 mmol), in W-methyl-2-pyrrolidone (1 mL). 2- Propenoy! chloride (0.021 mL, 0.258 mmol) was added and the reaction was stirred at room temperature for 1 hour. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) followed by a second purification by mass directed automated preparative HPLC (formic acid modifier) to afford the title compound (20 mg, 0.038 mmol, 29 % yield). LCMS (Method D): Rt 0.84 minutes; m/z 480 (MH+).
Example 59:
W -(1-acryloyl-4^iperidinyl)-6-{[c/*s-2,6-dim
[1 ,3]thiazolo[5,4-ft]pyridin-2-yl-2,4-pyrimidinediamine
Figure imgf000188_0002
A round bottom flask was charged with 1 , 1 -dimethylethyl 4-{[4-{[c/s-2,6-dimethyl-4- morpholinyl]methyl}-6-([1 ,3]thiazoio[5,4-0]pyridin-2-ylamino)-2-pyrimidinyl]amino}-1 - piperidinecarboxylate (148 mg, 0.267 mmol), 4M HCI in dioxane (2.63 mL, 10.51 mmo!), dichloromethane (3 mL) and methanol (2 mL). The reaction was stirred at room temperature for 1 hour. The solvent was evaporated to dryness. The residue was dissolved in W-methyl-2-pyrrolidone (3 mL) and treated with A/,W-diisopropylethy!amine (0.234 mL, 1 ,338 mmol) and 2-propenoyl chloride (0,043 mL, 0.535 mmol). The reaction was stirred at room temperature for 1 hour. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (56 mg, 0.11 mmol, 41 % yield). LCMS (Method D): Rt 0.86 minutes; m/z 509 (MH+).
Example 60:
W2-[(3S}-1-acryloyl-3-pyrrolidiny^6-{^
idin-2-yI-2,4-pyrimidinediamine
Figure imgf000189_0001
A round bottom flask was charged with 1 ,1-dimethylethyl (3S)-3-{[4-{[c/s-2,6-dimethyl-4- mo hol!nyl]methyl}-6-([1 ,3]thiazo!o[5,4-J!>]p ridin-2-yίamino)-2-pyπmidinyl]amino}-1- pyrrolidinecarboxylate (1 17 mg, 0.216 mmol), 4M HCI in dioxane (2.126 mL, 8.5 mmol), dichloromethane (3 mL) and methanol (2 mL). The reaction was stirred at room temperature for 1 hour. The solvent was evaporated to dryness. The residue was dissolved in V-methyi-2-pyrrolidone (2 mL) and treated with tyW-diisopropy!ethylamine (0.189 mL, 1.082 mmol) and 2-propenoyl chloride (0.035 mL, 0.433 mmol). The reaction was stirred at room temperature for 1 hour. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (52 mg, 0.11 mmol, 41 % yield) as an off-white solid. LCMS (Method D): Rt 0.84 minutes; m/z 495 (MH+). Example 61 :
A/-(2-{[4-{[c/s-2,6-dimethyl-4-mo
ylamino)-2-pyrimidinyl]amino}ethyi)-2-propenamide
Figure imgf000189_0002
A round bottom flask was charged with 1 , 1-dimethylethyl (2-{[4-{[c/s-2,6-dimethyl-4- morpholiny!]methy[}-6-([1 ,3]thiazolo[5,4-j ]pyridin-2-ylamino)-2- pyrimidiny!]amino}ethyl)carbamate (134 mg, 0.26 mmol), 4M HCI in dioxane (2.56 mL, 10.23 mmol), dichloromethane (3 mL) and methanol (2 mL), The reaction was stirred at room temperature for 1 hour. The solvent was evaporated to dryness. The residue was dissolved in A/-methyl-2-pyrrolidone (2 mL) and treated with W,A/-diisopropylethylamine (0.227 mL, 1.302 mmol) and 2-propenoyl chloride (0.042 mL, 0.521 mmol). The reaction was stirred at room temperature for 1 hour. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (59 mg, 0.126 mmo!, 48 % yield) as an off-white solid. LCMS (Method D); Rt 0.8 minutes; m/z 469 (MH+).
Example 62:
W-(1 -acryioyl-4-piperidiny!)-4-{[c/s-2,6-dim^
[1 ,3]thiazolo[5,4-Jb|pyridin-2-yl-2,6-pyridinediamine
Figure imgf000190_0001
A round bottom flask was charged with 1 ,1-dimethylethyl 4-{[4-{[(c/s)-2,6-dimethyl-4- morpholinyl]methyl}-6-([1 ,3]thiazolo[5,4-b]pyhdin-2-ylamino)-2-pyridinyl]amino}-1 - piperidinecarboxyiate (106 mg, 0.191 mmol), 4M HCI in dioxane (1 .881 mL, 7.52 mmol), dichloromethane (3 mL) and methanol (2 mL). The reaction was stirred at room temperature for 1 hour. The solvent was evaporated to dryness. The residue was dissolved in A/-methyl-2-pyrrolidone (2 mL) and treated with tyW-diisopropylethylamine (0.167 mL, 0.957 mmol) and 2-propenoyl chloride (0.031 mL, 0.383 mmol). The reaction was stirred at room temperature for 1 hour. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (36 mg, 0.071 mmol, 37 % yield) as an off-white solid. LCMS (Method D): Rt 0.93 minutes; m/z 508 (MH+).
Example 63:
W-[{3S)-1 -acryloyl-3-pyrrolidinyl]-4-fl^
[I .SJthiazolofS^-feJpyridin^-yi^.e-pyridinediamine
Figure imgf000191_0001
A round bottom flask was charged with 1 , 1-dimethylethy! (3S)-3-{[4-{[c/s-2,6-dimethyl-4- morpholinyl]methyl}-6-(t1 ,3]thiazolo[5,4-b]pyridin-2-ylamino)-2-pyridinyl]amino}-1- pyrro!idinecarboxylate (104 mg, 0.193 mmo!), 4M HCI in dioxane (1.893 mL, 7.57 mmo!), dichloromethane (3 mL) and methanol (2 mL). The reaction was stirred at room temperature for 1 hour. The solvent was evaporated to dryness. The residue was dissolved in A/-methyl-2-pyrrolidone (3 mL) and treated with A/.W-diisopropylethylamine (0.168 mL, 0.964 mmol) and 2-propenoyl chloride (0.031 mL, 0,385 mmoS). The reaction was stirred at room temperature for 1 hour. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (36 mg, 0.074 mmol, 38 % yield) as an off-white solid. LCMS (Method D): Rt 0.89 minutes; m/z 494 (MH+).
Example 64:
-aery loyl-3-piperidiny ^-ifc/s^je-dimethy -morphoMn lJmethyl}-/^- ridin-2-yl-2,6-pyridinediamine
Figure imgf000191_0002
A round bottom flask was charged with 1 ,1-dimethylethyl 3-{[4-{[c s-2,6-dimethyi-4- morpholinyl]methyi}-6-([1 ,3]thiazoio[5,4-jb]pyridin-2-ylamino)-2-pyridinyl]amino}-1 - piperidinecarboxylate (50 mg, 0.09 mmol), 4M HCI in dioxane (0.887 mL, 3.55 mmol), dichloromethane (3 mL) and methanol (2 mL). The reaction was stirred at room temperature for 1 hour. The solvent was evaporated to dryness. The residue was dissolved in A/-methyl-2-pyrroiidone (2 mL) and treated with W,W-diisopropylethylamine (0.079 mL, 0.451 mmol) and 2-propenoyl chloride (0.015 mL, 0.181 mmol). The reaction was stirred at room temperature for 1 hour. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (7 mg, 0.015 mmol, 16 % yield) as a yellow solid. LCMS (Method D): Rt 0.94 minutes; m/z 508 (MH+).
Example 65:
W-[(3S)-1-acryloyl-3-piperidinyl]-4-{[c/s-2,6-dimethyl-4-morpholinyl]m
ridin-2-yl-2,6-pyridinediamine
Figure imgf000192_0001
A microwave via! was charged with W-(6-chloro-4-((c/s-2,6- dimethylmorpholino)methyl)pyridin-2-yl)thiazolo[5,4-ij]pyridin-2-amine (50 mg, 0.128 mmol), 1 ,1-dimethylethyl (3S)-3-amino-1-piperidinecarboxylate (38.5 mg, 0.192 mmol), dicyclohexylt2,,4',6'-tris(1-methylethyl)-3,6-bis(methyloxy)-2-biphenylyl]phosphane
(BrettPhos) (6.9 mg, 0.013 mmol), chlorot2-(dicyclohexylphosphino)-3,6-dimethoxy-2'-4'- 6,-tri-i-propyl-1 ,1 '-biphenyl][2-(2-aminoethyl)phenyl]palladium(ll) (10.2 mg, 0.013 mmol) and tetrahydrofuran (1 mL). The system was sealed and placed under an atmosphere of nitrogen using a vacuum purge (3 times). Lithium bis(trimethylsiiyl)amide, 1 M in tetrahydrofuran (0.412 mL, 0.412 mmol) was added and the vial was heated at 80 °C in the preheated oil bath for 1.5 hours. The vial was cooled down to room temperature and the reaction mixture was partitioned between dichloromethane (10 mL) and saturated aqueous ammonium chloride (10 mL). After separation, the organic extract was dried using a hydrophobic frit and evaporated to dryness. The yellow residue was dissolved in dichloromethane (3 mL) and methanol (1 mL). The solution was treated with 4M HCl in dioxane (0.5 mL, 2 mmol) and stirred at room temperature for 16 hours. The solvent was evaporated to dryness. The yellow residue was dissolved in A/-methyl-2-pyrrolidone (1 mL) and W,W-diisopropylethy!amine (0.067 mL, 0.385 mmol). 2-Propenoyl chloride (0.021 mL, 0.256 mmol) was added and the reaction was stirred at room temperature for 1 hour. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (16 mg, 0.032 mmol, 25 % yield) as a brown solid. LCMS (Method D): Rt 0.94 minutes; m/z 508 (MH+).
Example 66: W-[(3f?)-1 -actyloyl-3-piperidinyl]-4-{[cis-2,6-dimethyl-4-morpholinyl]meth
ridin-2-yl-2,6-pyridinediamine
Figure imgf000193_0001
A microwave vial was charged with A/-(6-chloro-4-((c s-2,6- dimethylmorpholino)methyl)pyridin-2-yl)thiazolo[5,4-0]pyridin-2-amine (50 mg, 0.128 mmol), 1 ,1-dimethylethyl (3R)-3-amino-1 -piperidinecarboxylate (38.5 mg, 0.192 mmol) and {1 ,3-bis[2,6-bis(1-meihy!eihyl)phenyl]-2-imidazolidinyl}(chloro)(2-methyl-2-propen-1- yl)pailadium (22.64 mg, 0.038 mmol). The system was sealed and placed under an atmosphere of nitrogen using a vacuum purge. Lithium bis(trimethylsilyl)amide, 1 M in tetrahydrofuran (0.5 mL, 0.5 mmo!) was added. The vial was stirred in the preheated oi! bath at 70 °C for 1 hour. The vial was coo!ed down to room temperature and the reaction mixture was partitioned between dich!oromethane (10 mL) and saturated aqueous ammonium chloride (10 mL). After separation, the organic extract was dried using a hydrophobic frit and evaporated to dryness. The yeliow residue was dissolved in dichioromethane (3 mL) and methanol (1 mL). The solution was treated with 4 M HCI in dioxane (0.5 mL, 2 mmol) and stirred at room temperature overnight. The solvent was evaporated to dryness, The yellow residue was dissolved in W-methyl-2-pyrrolidone (1 mL) and W,A/-diisopropylethylamine (0.1 12 mL, 0.641 mmol). 2-Propenoyl chloride (0.021 mL, 0.256 mmol) was added and the reaction was stirred at room temperature for 1 hour. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (14 mg, 0.028 mmol, 21 % yield) as a brown solid. LCMS (Method D): Rt 0.94 minutes; m/z 508 (MH+).
Example 67:
W-{1-[(2E)-4-(dimethylamino)-2-butenoyl]-4-piperidiny[}-4-{[c/s-2(6-dim
morpholinyi]methyl}-AT-[1 ,3]thiazolo[5,4-ft]pyrid!n-2-yl-2,6-pyridinediamin
Figure imgf000194_0001
A round bottom flask was charged with 1 , 1 -dimethy!ethyi ^{^-{[c s^^-dimethyl^- morpholiny!]methyl}-6-([1 ,3]thiazolo[5,4-/b3pyridin-2-ylamino)-2-pyridinyl]amino}-1 - piperidinecarboxylate (106 mg, 0.191 mmol), 4M HCI in dioxane (1 .881 mL, 7.52 mmol), dichloromethane (3 mL) and methanol (2 mL). The reaction was stirred at room temperature for 1 hour. The solvent was evaporated to dryness. The residue was dissolved in W-methyl-2-pyrrolidone (2 mL) and treated with /V,A/-diisopropylethylamine (0.1 mL, 0.574 mmol). This solution was added to a stirred mixture of 0-(7- azabenzotriazol-1-y!)-Ay,Ay,/\/' A/'-tetramethyluronium hexafiuorophosphate (109 mg, 0.287 mmol), (2£)-4-(dimethylamino)-2-butenoic acid hydrochloride (47.6 mg, 0.287 mmol) and WJV-diisopropylethy!amine (0.1 mL, 0.574 mmol) in W-methyi-2-pyrrolidone (2 mL). The reaction was stirred at room temperature for 1 hour. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (60 mg, 0.107 mmol, 56 % yield) as an off-white solid. LCMS (Method D): Rt 0.92 minutes; m/z 565 (MH+).
Example 68:
AH(3S)-1-[(2E)-4-(dimethylamino)-2-butenoy
Figure imgf000194_0002
A round bottom flask was charged with 1 ,1 -dimethylethyl (3S)-3-{[4-{[c/'s-2,6-dimethyl-4- morpholinyl]methyl}-6-(E1 ,3]thiazolo[5,4-0]pyridin-2-ylamino)-2-pyridinyl]amino}-1- pyrrolidinecarboxyiate (104 mg, 0.193 mmol), 4M HCI in dioxane (1.893 mL, 7.57 mmol), dichloromethane (3 mL) and methanol (2 mL). The reaction was stirred at room temperature for 1 hour. The solvent was evaporated to dryness. The residue was dissolved in v-methyl-2-pyrrolidone (2 mL) and treated with Λ/,/V-diisopropylethylamine (0.101 mL, 0.578 mmo!).This solution was added to a stirred mixture of 0-(7- azabenzotriazol-1-yl)-A/,A/,/V',A/'-tetramethyluronium hexafiuorophosphate (1 10 mg, 0.289 mmol), (2£)-4-(dimethylamino)-2-butenoic acid hydrochloride (48 mg, 0.289 mmol) and Λ/,/V-diisopropylethylamine (0.101 mL, 0.578 mmol) in A/-methyl-2-pyrroiidone (2 mL). The reaction was stirred at room temperature for 1 hour. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (47 mg, 0,085 mmol, 44 % yield) as an off-white solid. LCMS (Method D): Rt 0.89 minutes; m/z 551 (MH+).
Example 69:
W-{1 -n2/E -4-(dimethylamino)-2-butenoyl]-3-piperidinyl}-4-{|c/s-2,6-dimeth
morpholinyllmethylJ-W'-tl .SJthiazolotS^-^pyridin^-yl^.e-pyridinediamine
Figure imgf000195_0001
A round bottom flask was charged with 1 ,1 -dimethylethyi 3-{[4-{[c s-2,6-dimethyl-4- morpholsnyllmethy -B-itl .SjthiazoloES^-bJpyridin^-yiaminoJ^-pyridinyllamino}-^ piperidinecarboxylate (50 mg, 0.09 mmol), 4 HCI in dioxane (0.887 mL, 3.55 mmol), dich!oromethane (3 mL) and methanol (2 mL). The reaction was stirred at room temperature for 1 hour. The solvent was evaporated to dryness. The residue was dissolved in A/-methyl-2-pyrrolidone (2 mL) and treated with A/JV-diisopropylethylamine (0.047 mL, 0.271 mmol). This solution was added to a stirred mixture of 0-(7- azabenzotriazol-l-y -A/^^' / '-tetramethyluronium hexafiuorophosphate (51.5 mg, 0.135 mmol), (2-_r)-4-(dimethylamino)-2-butenoic acid hydrochloride (22.43 mg, 0.135 mmol) and A/JV-diisopropy!ethylarnine (0.047 mL, 0.271 mmol) in /V-methyl-2-pyrrolidone (2 mL). The reaction was stirred at room temperature for 1 hour. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (9 mg, 0.016 mmo!, 18 % yie!d) as a yellow solid. LCMS (Method D): Rt 0.92 minutes; m/z 565 (MH+).
Example 70:
formic acid - (2E)-4-(dimethy!amino)-W-(2-{l4-(4-morphoIinylmethyl)-6- (t1,3]thiazolo[5,4- 53pyridin-2-ylamino)-2-pyrimidinyl]amino}ethyl)-2-butenamide
Figure imgf000196_0001
A mixture of (2£)-4-(dimethylamino)-2-butenoic acid hydrochloride (35 mg, 0.21 mmoi), A/,/V-diisopropylethylamine (0.73 ml_, 0.42 mmoi) and 0-(7-azabenzotriazol-1-yi)- Λ/,Λ/,Λ/',Λ/'-tetramethyluronium hexafluorophosphate (80 mg, 0.210 mmoi) in /V-methyi-2- pyrrolidone (1 mL) was stirred at room temperature for 2 minutes. A solution of Λ/ -(2- aminoethyi)-6-(4-morpho!inylmethyl)-/\/''-[1 ,3]thiazoloi5,4-/)]pyridin-2-yi-2,4- pyrimidinediamine dihydrochloride (64 mg, 0.14 mmoi) and Λ/,/V-diisopropylethylamine (0.73 mL, 0.42 mmo!) in W-methyl-2-pyrrolidone (1 mL) was added to the previous solution and the reaction was stirred at room temperature for 1 hour. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) followed by a second purification by mass directed automated preparative HPLC (formic acid modifier) to afford the title compound (25 mg, 0.042 mmoi, 30 % yield). LCMS (Method D): Rt 0.73 minutes; m/z 498 (MH+).
Example 71 :
W2-{1-[(2E)-4-(dimethylamino)-2-butenoy
[1 ,33thiazolo[5,4-f)]pyridin-2-yl-2,4-pyrimidinediamine
Figure imgf000197_0001
A mixture of (2£)-4-(dimethylamino)-2-butenoic acid hydrochloride (35 mg, 0.21 mmol), W,W-diisopropySethy!amine (0.73 mL, 0.42 mmol) and 0-(7-azabenzotriazol-1-yl)- Λ/,Λ/,Λ/' Λ/'-tetramethyluronium hexafluorophosphate (80 mg, 0.210 mmol) in /V-methyl-2- pyrrolidone (1 mL) was stirred at room temperature for 2 minutes. A solution of 6-(4- morpho!inylmethyl)- \/2-3-piperidinyl-A/4-[1 ,3]thiazolo[5,4-0]pyridin-2-yl-2,4- pyrimidinediamine dihydrochloride (70 mg, 0.14 mmol) and Λ/,/V-diisopropylethylamine (0,73 mL, 0.42 mmol) in A/-methyl-2-pyrro!idone (1 mL) was added to the previous solution and the reaction was stirred at room temperature for 1 hour. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (35 mg, 0.065 mmol, 46 % yield). LC S (Method D): Rt 0.78 minutes; m/z 538 (MH+).
Example 72:
W-{(3S)-1 -[{2£)-4-(dimethylamino)-2-butenoyl]-3-pyrrolidinyl}-4-(4- morpholinylmethyl)- -[1 ,3]thiazolo[5,4- >]pyridin-2-yl-2,6^yridinediamine
Figure imgf000197_0002
A round bottom flask was charged with 1 ,1 -dimethylethyl (3S)-3-{[4-(4-morpholinyimethyl)- 6-([1 ,3]thiazolo[5,4-i 3pyridin-2-yiamino)-2-pyridinyl]amino}-1-pyrrolidinecarboxylate (70 mg, 0.137 mmol), 4M HC! in dioxane (0.342 mL, 1.368 mmol), dichloromethane (3 mL) and methanol (1 mL). The reaction was stirred at room temperature for 16 hours. The reaction mixture was evaporated to dryness. The residue was dissolved in A/-methyl-2- pyrrolidone (1 ml) and W,W-diisopropylethylamine (0.072 mL, 0.41 mmo!) was added. The solution was added to a stirring solution of (2E)-4-(dimethylamino)-2-butenoic acid hydrochloride (34 mg, 0.205 mmol), 0-(7-azabenzotriazo!-1-yl)-W,W,W,W- tetramethyluronium hexaf!uorophosphate (78 mg, 0.205 mmoi), N,N- diisopropyiethylamine (0.072 mL, 0.410 mmoi) and /V-methy!-2-pyrrolidone (1 ml). The reaction was stirred at room temperature for 1 hour. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (33 mg, 0.063 mmol, 46 % yield). LCMS (Method D): Rt 0.81 minutes; m/z 523 (MH+).
Example 73:
/V-{1 -[(2E)-4-(dimethylamino)-2-butenoyl]-4-piperidinyl}-4-(4-morpholinyl^
-yl-2,6-pyridinediamine
Figure imgf000198_0001
A round bottom flask was charged with 1 , 1-dimethylethyl 4-{[4-(4-morpholinylmethyl)-6- ([1 ,3]thiazolo[5,4-0]pyridin-2-ylamino)-2-pyridinyl]amino}-1-plperidinecarboxylate (72 mg, 0.137 mmol), 4M HCI in dioxane (0.342 mL, 1.370 mmol), dichloromethane (3 mL) and methanol (1 mL). The reaction mixture was stirred at room temperature for 16 hours. The solvent was evaporated to dryness. The solid residue was dissolved in W-methyl-2- pyrrolidone (1 mL) treated with /V,W-diisopropylethylamine (0.072 mL, 0.411 mmol). This solution was added to a stirred mixture of 0-(7-azabenzotriazol-1 -yl)-W,/V,/V' /\/'- tetramethyluronium hexafluorophosphate (78 mg, 0.205 mmol), (2£)-4-(dimethylamino)-2- butenoic acid hydrochloride (34 mg, 0.205 mmol) and W,W-diisopropylethylamine (0.072 mL, 0.41 1 mmol) in /V-methyi-2-pyrrolidone (1 mL). The reaction was stirred at room temperature for 1 hour. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (37 mg, 0.069 mmol, 50 % yield) as a light brown solid. LCMS (Method D): Rt 0.83 minutes; m/z 537 (MH+). Example 74: N-{1 -[(2E)-4-(dimethylamino)-2-buteno
yridin-2-yl-2,6-pyridinediamine
Figure imgf000199_0001
A round bottom flask was charged with 1 ,1 -dimethylethyl 3-{[4-(4-morpholinylmethyl)-6- (t1 ,3]thiazolo[5,4-b]pyridin-2-ylamino)-2-pyridinyl]amino}-1-piperidinecarboxylate (80 mg, 0.152 mmol), 4M HCI in dioxane (0.38 mL, 1.522 mmol), dichloromethane (3 mL) and methanol (1 mL). The reaction mixture was stirred at room temperature for 16 hours. The solvent was evaporated to dryness. The solid was dissolved in W-methyl-2-pyrrolidone (1 mL) and treated with tyW-ditsopropylethylamine (0.08 mL, 0.457 mmol), This solution was added to a stirred mixture of O-iy-azabenzotriazol-l-y -^A/^' /V-tetramethyluronium hexafluorophosphate (87 mg, 0.228 mmol), (2£)-4-(dimethylamino)-2-butenoic acid hydrochloride (37,8 mg, 0.228 mmol) and W,W-diisopropylethylamine (0.08 mL, 0.457 mmol) in W-methyi-2-pyrrolidone (1 mL). The reaction was stirred at room temperature for 1 hour. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (40 mg, 0.075 mmol, 49 % yield) as a light brown solid. LCMS (Method D): Rt 0.82 minutes; m/z 537 (MH+).
Example 75:
-aery loyI-4-piperidinyl)-4-(1 -piperidinylmethyl)-W -[1 ,3]thiazo!o[5,4- )]pyridin-2- -2,6-pyridinediamine
Figure imgf000199_0002
A solution of 1 , 1 -dimethylethyl 4-{[4-formyl-6-([1 ,3]thiazolo[5,4-b]pyridin-2-ylamino)-2- pyridinyl]amino}-1-piperidinecarboxylate (50 mg, 0.1 1 mmol) in tetrahydrofuran (0.5 mL) was treated with piperidine (0.022 mL, 0,22 mmol) and the reaction mixture was stirred for 15 minutes. The mixture was then treated with dichloromethane (2 mL) and sodium triacetoxyborohydride (46.6 mg, 0.22 mmo!). The reaction mixture was stirred for 16 hours. Saturated aqueous sodium bicarbonate (1 mL) was added and the reaction was stirred for 30 minutes. Dichloromethane (5 mL) was added and the phases were separated. The organic extract was dried using a hydrophobic frit and evaporated to dryness. A round bottom flask was charged with the obtained solid, 4 HCI in dioxane (0.938 mL, 3.75 mmol), dichloromethane (3 mL) and methanol (2 mL). The reaction was stirred at room temperature for 1 hour. The solvent was evaporated to dryness. The residue was dissolved in A/-methyl-2-pyrrolidone (1 mL) and treated with N,N- diisopropylethylamine (0.083 mL, 0.477 mmol) and 2-propenoyl chloride (0.015 mL, 0.191 mmol). The reaction was stirred at room temperature for 1 hour. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (32 mg, 0.068 mmo!, 62 % yield) as a light brown solid. LC S (Method D): Rt 1.00 minutes; m/z 478 (MH+).
Example 76:
-acryloyl-4-piperidinyl)-W-[1 ,3]thiazolo[5,4-i)3pyndin-2-yl-4-({[(1 S)-1 ,2,2- trimethylpropyl]amino}methyl)-2,6-pyridinediamine
Figure imgf000200_0001
A solution of 1 , 1 -dimethylethyl 4-{[4-formyl-6-([1 ,3]thiazolo[5,4-b]pyridin-2-ylamino)-2- pyridinyl3amino}-1-piperidinecarboxy!ate (50 mg, 0.1 1 mmol) in tetrahydrofuran (0.5 mL) was treated with [(I SJ-l ^^-trimethyipropyijamine ( 22.5 mg, 0.22 mmol) and the reaction mixture was stirred for 15 minutes. The mixture was then treated with dichloromethane (2 mL) and sodium triacetoxyborohydride (46.6 mg, 0.22 mmol). The reaction mixture was stirred for 16 hours. Saturated aqueous sodium bicarbonate (1 mL) was added and the reaction was stirred for 30 minutes. Dichloromethane (5 mL) was added and the phases were separated. The organic extract was dried using a hydrophobic frit and evaporated to dryness. A round bottom flask was charged with the obtained solid, 4M HCI in dioxane (1.081 mL, 4.33 mmol), dichloromethane (3 mL) and methanol (2 mL). The reaction was stirred at room temperature for 1 hour. The solvent was evaporated to dryness. The residue was dissolved in /V-methyl-2-pyrrolidone (1 mL) and treated with N,N- diisopropylethylamine (0.096 mL, 0.55 mmol) and 2-propenoyl chloride (0.018 mL, 0.22 mmol). The reaction was stirred at room temperature for 1 hour. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (15 mg, 0.03 mmol, 27 % yield) as a light brown solid. LC S (Method D): Rt 1.1 1 minutes; m/z 494 (MH+).
Example 77:
W-(1-actyloyl-4-piperidinyl)-Ar-[1 ,3]thiazolo[5,4- j]pyridin-2-yl-4-{{[(1R)-1 ,2,2- trimethylpropyl]amino}methyl)-2,6-pyridinediamine
Figure imgf000201_0001
A solution of 1 ,1-dimethylethyl 4-{[4-formyl-6-([1 ,3]thiazolo[5,4-b]pyridin-2-ylamino)-2- pyridinyl]amino}-1-piperidinecarboxylate (50 mg, 0.1 1 mmol) in tetrahydrofuran (0.5 mL) was treated with [(1 f?)-1 ,2,2-trimethylpropyl]amine (22.5 mg, 0,22 mmoi) and the reaction mixture was stirred for 15 minutes. The mixture was then treated with dichloromethane (2 mL) and sodium triacetoxyborohydride (46.6 mg, 0.22 mmol). The reaction mixture was stirred for 16 hours. Saturated aqueous sodium bicarbonate (1 mL) was added and the reaction was stirred for 30 minutes. Dichloromethane (5 mL) was added and the phases were separated. The organic extract was dried using a hydrophobic frit and evaporated to dryness. A round bottom flask was charged with the obtained solid, 4 HCI in dioxane (1.081 mL, 4.33 mmol), dichloromethane (3 mL) and methanol (2 mL). The reaction was stirred at room temperature for 1 hour. The solvent was evaporated to dryness. The residue was dissolved in A/-methyl-2-pyrroltdone (1 mL) and treated with N,N- diisopropylethylamine (0.096 mL, 0.55 mmol) and 2-propenoyl chloride (0.018 mL, 0.22 mmol). The reaction was stirred at room temperature for 1 hour. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (1 1 mg, 0.023 mmoi, 21 % yield) as a light brown solid. LCMS (Method D): Rt 1 .1 1 minutes; m/z 494 (MH+).
Example 78:
W-(1-acryloyl-4-piperidinyl)-4-[(2-methyl-1 -pyrrol idinyl}methyl]-W-[1,3]thiazolo[5,4- Jb3pyridin-2-yi-2,6-pyridinediamine
Figure imgf000202_0001
A solution of 1 , 1-dimethylethyl 4-{[4-formyl-6-([1 ,3]thiazolo[5,4-/!?]pyndin-2-ylamino)-2- pyridinyl]amino}-1 -piperidinecarboxy!ate (50 mg, 0.1 1 mmol) in tetrahydrofuran (0.5 mL) was treated with 2-methylpyrrolidine (19.5 mg, 0.22 mmol) and the reaction mixture was stirred for 15 minutes. The mixture was then treated with dichioromethane (2 mL) and sodium triacetoxyborohydride (46.6 mg, 0.22 mmol). The reaction mixture was stirred for 16 hours. Saturated aqueous sodium bicarbonate (1 mL) was added and the reaction was stirred for 30 minutes. Dichioromethane (5 mL) was added and the phases were separated. The organic extract was dried using a hydrophobic frit and evaporated to dryness. A round bottom flask was charged with the obtained solid, 4M HCI in dioxane (1.013 mL, 4.05 mmol), dichioromethane (3 mL) and methanol (2 mL). The reaction was stirred at room temperature for 1 hour. The solvent was evaporated to dryness. The residue was dissolved in W-methyl-2-pyrrolidone (1 mL) and treated with N,N- diisopropylethylamine (0.09 mL, 0.516 mmol) and 2-propenoyl chloride (0.017 mL, 0.206 mmol). The reaction was stirred at room temperature for 1 hour. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (26 mg, 0.054 mmol, 49 % yield) as a light brown solid. LCMS (Method D): Rt 0.99 minutes; m/z 478 (MH+). Example 79:
W-(1 -acryloyl-4-piperidinyi)-4-[(2,2-dimethyl-4-morpholinyl)methyl3-W'- [1 ,3]thiazolo[5,4-fe]pyridin-2-yl-2,6-pyridinediamine
Figure imgf000202_0002
A solution of 1 , 1 -dimethylethyl 4-{[4-formyi-6-([1 ,3]thiazolo[5,4-/}]pyridin-2-ylamino)-2- pyridinyl]amino}-1 -pipendinecarboxylate (50 mg, 0.1 1 mmol) in tetrahydrofuran (0.5 mL) was treated with 2,2-dimethylmorpho!ine (25.3 mg, 0.22 mmol) and the reaction mixture was stirred for 15 minutes. The mixture was then treated with dichloromethane (2 mL) and sodium tnacetoxyborohydride (46.6 mg, 0.22 mmol). The reaction mixture was stirred for 16 hours. Saturated aqueous sodium bicarbonate (1 mL) was added and the reaction was stirred for 30 minutes. Dichloromethane (5 mL) was added and the phases were separated. The organic extract was dried using a hydrophobic frit and evaporated to dryness. A round bottom flask was charged with the obtained solid, 4 HCI in dioxane (1.072 mL, 4.29 mmol), dichloromethane (3 mL) and methanol (2 mL). The reaction was stirred at room temperature for 1 hour. The solvent was evaporated to dryness. The residue was dissolved in /V-methyl-2-pyrrolidone (1 mL) and treated with N,N- diisopropylethyiamine (0.095 mL, 0.545 mmol) and 2-propenoyl chloride (0.018 mL, 0.218 mmol). The reaction was stirred at room temperature for 1 hour. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (27 mg, 0.053 mmol, 48 % yield) as a light brown solid. LCMS (Method D): Rt 0.99 minutes; m/z 508 (MH+).
Example 80:
/V-ttSSJ-l-acryloyl-S-pyrrolidinyll^-il-piperidinylmethy -Af-II ^SthiazoIoIS^- Jb]pyridin-2-yl-2,6-pyridinediamine
Figure imgf000203_0001
A solution of 1 ,1 -dimethylethyl (3S)-3-{[4-formyl-6-(t1 ,33thiazolo[5,4-0]pyridin-2-ylamino)- 2-pyridinyl]amino}-1-pyrrolidinecarboxylate (50 mg, 0.1 14 mmol) in tetrahydrofuran (0.5 mL) was treated with piperidine (0.023 mL, 0.227 mmol) and the reaction mixture was stirred for 15 minutes. The mixture was then treated with dichloromethane (2 mL) and sodium tnacetoxyborohydride (48.1 mg, 0.227 mmo!). The reaction mixture was stirred for 16 hours. Saturated aqueous sodium bicarbonate (1 mL) was added and the reaction was stirred for 30 minutes. Dichloromethane (5 mL) was added and the phases were separated. The organic extract was dried using a hydrophobic frit and evaporated to dryness. A round bottom flask was charged with the obtained solid, 4M HCI in dioxane (1.177 mL, 4.71 mmol), dichloromethane (3 mL) and methanol (2 mL). The reaction was stirred at room temperature for 1 hour. A further portion of 4 HC! in dioxane (1.177 mL, 4.71 mmol) was added and the reaction was stirred for 30 minutes. The solvent was evaporated to dryness. The residue was dissolved in A/-methyl-2-pyrrolidone (1 mL) and treated with W,/V-diisopropyiethylamine (0.103 mL, 0.589 mmol) and 2-propenoyl chioride (0.019 mL, 0.235 mmol). The reaction was stirred at room temperature for 1 hour. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (29 mg, 0.063 mmol, 55 % yield) as a light brown solid. LCMS (Method D): Rt 0.98 minutes; m/z 464 (MH+). Example 81 :
W-[(3S)-1-acryloyl-3-pyrrolidinyl]-4-[(2(2-dimethyl-4-morpholiny!)methyl]-W'- [1 ,3]thiazolo[5,4-i)3pyridin-2-yl-2,6-pyridinediamine
Figure imgf000204_0001
A solution of 1 ,1 -dimethyiethyl (3S)-3-{[4-formyl-6-([1 ,3]thiazo!o[5,4-/b]pyridin-2-ylamino)- 2-pyridinyi]amino}-1-pyrrolidinecarboxy!ate (50 mg, 0.114 mmol) in tetrahydrofuran (0.5 mL) was treated with 2,2-dimethylmorpholine (26.1 mg, 0.227 mmol) and the reaction mixture was stirred for 15 minutes. The mixture was then treated with dichloromethane (2 mL) and sodium triacetoxyborohydride (48.1 mg, 0.227 mmol). The reaction mixture was stirred for 16 hours. Saturated aqueous sodium bicarbonate (1 mL) was added and the reaction was stirred for 30 minutes. Dichloromethane (5 mL) was added and the phases were separated. The organic extract was dried using a hydrophobic frit and evaporated to dryness. A round bottom flask was charged with the obtained solid, 4M HCI in dioxane (1.186 mL, 4.74 mmol), dichloromethane (3 mL) and methanol (2 mL). The reaction was stirred at room temperature for 1 hour. A further portion of 4M HCI in dioxane (1.186 mL, 4.74 mmol) was added and the reaction was stirred for 30 minutes. The solvent was evaporated to dryness. The residue was dissolved in W-methyl-2-pyrrolidone (1 mL) and treated with V,/V-diisopropylethy!amine (0.104 mL, 0.593 mmol) and 2-propenoyl chloride (0.019 mL, 0.237 mmol). The reaction was stirred at room temperature for 1 hour. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (27 mg, 0.055 mmol, 48 % yield) as a light brown solid. LCMS (Method D): Rt 0.95 minutes; m/ 494 (MH+).
Example 82:
Λ/-[{3S)-1-ac^γloyl-3- yrrolidiny!j-4-[(4-methyl-1 -piperazin I)methyl]-Λ/,- -i)3pyridin-2-yi-2,6-pyridirtediamfne
Figure imgf000205_0001
A solution of 1 , 1 -dimethylethyi (3S)-3-{[4-formyl-6-([1 ,3]thiazolo[5,4-jb]pyridin-2-ylamino)- 2-pyridinyl]amino}-1-pyrrolidinecarboxylate (50 mg, 0.114 mmo!) in tetrahydrofuran (0.5 mL) was treated with 1-methylpiperazine (0.025 mL, 0.227 mmol) and the reaction mixture was stirred for 15 minutes. The mixture was then treated with dichloromethane (2 mL) and sodium triacetoxyborohydride (48.1 mg, 0.227 mmol). The reaction mixture was stirred for 6 hours. Saturated aqueous sodium bicarbonate (1 mL) was added and the reaction was stirred for 30 minutes. Dichloromethane (5 mL) was added and the phases were separated. The organic extract was dried using a hydrophobic frit and evaporated to dryness. A round bottom flask was charged with the obtained solid, 4M HCI in dioxane (1.201 mL, 4.8 mmol), dichloromethane (3 mL) and methanol (2 mL). The reaction was stirred at room temperature for 1 hour. A further portion of 4M HCI in dioxane (1.201 mL, 4.8 mmol) was added and the reaction was stirred for 30 minutes. The solvent was evaporated to dryness. The residue was dissolved in A/-methyl-2-pyrrolidone (1 mL) and treated with /V,W-diisopropyiethylamine (0.105 mL, 0.6 mmol) and 2-propenoyl chloride (0.019 mL, 0.24 mmol). The reaction was stirred at room temperature for 1 hour. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (32 mg, 0.067 mmol, 59 % yield) as a Sight brown solid. LCMS (Method D): Rt 0.78 minutes; m/z 479 (MH+).
Example 83:
W-[(3S)-1 -actYloyl-3-pyrroiidi nyl]-^ ^
trimethy!propyl]amino}methyl}-2,6-pyridinediamine
Figure imgf000206_0001
A solution of 1 , 1-dimethyiethyl (3S)-3-{[4-formyl-6-([1 ,3]thiazolo[5,4-/)]pyridin-2-ylamino)- 2-pyridinyl]amino}-1 -pyrroiidinecarboxylate (50 mg, 0.1 14 mmol) in tetrahydrofuran (0.5 mL) was treated with [(1 R)-1 ,2,2-trimethylpropy!]amine (0.031 mL, 0.227 mmol) and the reaction mixture was stirred for 15 minutes. The mixture was then treated with dichioromethane (2 mL) and sodium triacetoxyborohydride (48.1 mg, 0.227 mmol). The reaction mixture was stirred for 4 hours. A second portion of (2R)-3,3-dimethyl-2- butanamine (0.016 mL, 0.114 mmol) and sodium triacetoxyborohydride (96.2 mg, 0.454 mmoi) was added and the reaction was stirred at room temperature for 14 hours. Saturated aqueous sodium bicarbonate (1 mL) was added and the reaction was stirred for 30 minutes. Dichioromethane (5 mL) was added and the phases were separated. The organic extract was dried using a hydrophobic frit and evaporated to dryness. A round bottom flask was charged with the obtained solid, 4M HCI in dioxane (1.121 mL, 4.49 mmol), dichioromethane (3 mL) and methanol (2 mL). The reaction was stirred at room temperature for 1 hour. A further portion of 4M HCi in dioxane (1.201 mL, 4.8 mmol) was added and the reaction was stirred for 1.5 hours. The solvent was evaporated to dryness. The residue was dissolved in A/-methyl-2-pyrroiidone (1 mL) and treated with N,N- diisopropylethy!amine (0.1 mL, 0.571 mmol) and 2-propenoyl chloride (0.011 mL, 0.137 mmol). The reaction was stirred at room temperature for 1 hour. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (42 mg, 0.088 mmol, 77 % yield) as a light brown solid, LCMS (Method D): Rt 1.07 minutes; m/z 480 (MH+).
Example 84:
W-[(3S)-1 -acryloyl-3-pyrrolidinyl]-W-[1 ,3]thiazolo[5t4-J ]pyridin-2-yl-4-({[(1 S)-1 ,2,2- trimethylpropyl3amino}methyl)-2,6-pyridinediamine
Figure imgf000207_0001
Method A
A solution of 1 , 1-dimethylethy! (3S)-3-{[4-formyi-6-([1 ,3]thiazolo[5,4-/b]pyridin-2-ylamino)- 2-pyridinyl]amino}-1-pyrroIidinecarboxy!ate (100 mg, 0.227 mmol) in tetrahydrofuran (1 mL) was treated with [(1 S)-1 ,2,2~trimethylpropyl]amine (0.061 mL, 0.454 mmol) and the reaction mixture was stirred for 15 minutes. The mixture was then treated with dichioromethane (4 mL) and sodium triacetoxyborohydride (96 mg, 0.454 mmol). The reaction mixture was stirred for 16 hours. A second portion of sodium triacetoxyborohydride (96 mg, 0.454 mmol) was added and the reaction was stirred at room temperature for 2 hours. Saturated aqueous sodium bicarbonate (2 mL) was added and the reaction was stirred for 30 minutes. Dichioromethane (5 mL) was added and the phases were separated. The organic extract was dried using a hydrophobic frit and evaporated to dryness. A round bottom flask was charged with the obtained solid, 4M HCl in dioxane (2.224 mL, 8.9 mmol), dichioromethane (3 mL) and methanol (2 mL). The reaction was stirred at room temperature for 1 .5 hours. The solvent was evaporated to dryness. The residue was dissolved in W-methyl-2-pyrrolidone (1 mL) and treated with Λ/JV-diisopropylethylamine (0.198 mL, 1.132 mmol) and 2-propenoyl chloride (0.022 mL, 0.272 mmol). The reaction was stirred at room temperature for 1 hour. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (50 mg, 0.104 mmol, 46 % yield) as a light brown solid. LCMS (Method D): Rt 1 .07 minutes; m/z 480 (MH+).
Method B
W- Sj-S-Pyrrolidinylj-W-EI ,3]thiazolo[5,4-jb]pyridin-2-yl-4-({[(1 S)-1 ,2,2-trimethy!propyl] amino}methyl)-2,6-pyridinediamine (100 mg, 0.235 mmol) was suspended in /V-methyl-2- pyrrolidone (2 mL) and stirred at 0 °C. Triethylamine (0.065 mL, 0.470 mmol) was added foliowed by 2-propenoyl chioride (0.022 mL, 0.258 mmol) and the reaction was stirred at 0 °C for 30 mintues. The reaction mixture was partitioned between dichloromethane:methano! (1 : 1 , 20mL) and saturated aqueous sodium carbonate (20mL). After separation, the organic extract was dried using a hydrophobic frit and concentrated under reduced pressure. The residue was subjected to purification by mass directed automated preparative HPLC (trifluoroacetic acid modifier) to afford, after basification with concentrated aqueous ammonia, the titie compound (46 mg, 41 %) as a white solid. LCMS (Method E): Rt 0.68 minutes; m/z 480 (MH+). 1H NMR (400 MHz, DMSO-d6> 120 °C) d 8.26 (dd, J = 1.4, 4.7 Hz, 1 H), 7.82 (dd, J = 1 .4, 8.1 Hz, 1 H), 7.32 (dd, J = 4.7, 8.1 Hz, 1 H), 6.62 - 6.46 (m, 2H), 6.41 (s, 1 H), 6.23 (s, 1 H), 6.12 (dd, J = 2.1 , 16.7 Hz, 1 H), 5.61 (dd, J = 2.1 , 10.1 Hz, 1 H), 4.76 - 4.65 (m, 1 H), 4.03 - 3.83 (m, 1 H), 3.76 - 3.68 (m, 1 H), 3.73 (d, J = 14.4 Hz, 1 H), 3.67 - 3.58 (m, 1 H), 3.54 (d, J - 14,4 Hz, 1 H), 3.49 (dd, J = 4.5, 1 1.6 Hz, 1 H), 2.38 - 2.26 (m, 1 H), 2.30 (q, J = 6.3 Hz, 1 H), 2.06 - 1.95 (m, 1 H), 0.99 (d, J = 6.3 Hz, 3H), 0.91 (s, 9H).
Example 85:
W-{1 -acryloyl-4-piperidinyl)-4-[(dimethylamino)^
-yl-2,6-pyridinediamine
Figure imgf000208_0001
A solution of 1 , 1-dimethylethyl 4-{[4-formyl-6-([1 ,3]thiazoio[5,4-t)]pyridin-2-y[amino)-2- pyridinyl]amino}-1-piperidinecarboxylate (48 mg, 0.106 mmol) in tetrahydrofuran (0.5 mL) was treated with dimethylamine, 2 in tetrahydrofuran (0.106 mL, 0.21 1 mmol) and the reaction mixture was stirred for 15 minutes. The mixture was then treated with dichloromethane (2 mL) and sodium triacetoxyborohydride (44.8 mg, 0.21 1 mmol). The reaction mixture was stirred for 16 hours. Saturated aqueous sodium bicarbonate (1 mL) was added and the reaction was stirred for 30 minutes. Dichloromethane (5 mL) was added and the phases were separated. The organic extract was dried using a hydrophobic frit and evaporated to dryness. A round bottom flask was charged with the obtained solid, 4M HCI in dioxane (1 .016 mL, 4.06 mmol), dichloromethane (3 mL) and methanol (2 mL). The reaction was stirred at room temperature for 1 .5 hours. The solvent was evaporated to dryness. The residue was dissolved in A/-methyl-2-pyrroiidone (1 mL) and treated with Λ/Jv-diisopropylethylamine (0.09 mL, 0.517 mmol) and 2-propenoyl chloride (0.017 mL, 0.207 mmol). The reaction was stirred at room temperature for 1 hour. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (8 mg, 0.018 mmol, 17 % yield) as a yellow solid. LCMS (Method D): Rt 0.86 minutes; m/z 438 (MH+). Example 86:
Λ/-[(35)-1 -3θΓγΙογΙ-3-ρϊρβπάϊηνΙ]-4-{ ^θΓρΗοΜηνΐΓίΐθΙΗγΙ)-ΛΓ-[1 ,3]ίΗί3Ζθ[ο[5,4- Jb]pyridin-2-yl-2,6-pyridinediamine
Figure imgf000209_0001
A microwave vial was charged with A/-(6-chloro-4-(morpholinomethyl)pyridin-2- yl)thiazolo[5,4-jb]pyridin-2-amine (75 mg, 0.207 mmol), 1 , 1 -dimethylethyl (3S)-3-amino-1- piperidinecarboxylate (62.3 mg, 0.31 1 mmol), dicyclohexyl[2',4',6'-tris(1-methylethyl)-3,6- bis(methy!oxy)-2-biphenylyljphosphane (BrettPhos) (1 1.13 mg, 0.021 mmol), chloro[2- (dicyclohexylphosphinoJ-S.e-dimethoxy-Z-^-e'-tri-i-propyl-l , 1 '-biphenylj[2-(2- aminoethy!)phenyl]palladium(ll) (16.56 mg, 0.021 mmol) and tetrahydrofuran (1 mL). The system was sealed and placed under an atmosphere of nitrogen using a vacuum purge (3 times). Lithium bis(trimethylsilyl)amide, 1 M in tetrahydrofuran (0.622 mL, 0.622 mmol) was added and the vial was heated at 80 °C in the preheated oil bath for 14 hours. The vial was cooled down to room temperature and the reaction mixture was partitioned between dichloromethane (10 mL) and saturated ammonium chloride (10 mL). After separation, the organic extract was dried using a hydrophobic frit and evaporated to dryness. The yellow residue was dissolved in dichloromethane (3 mL) and methanol (1 mL). The soiution was treated with 4M HCi in dioxane (0.5 mL, 2 mmol) and stirred at room temperature overnight. The solvent was evaporated to dryness. The yellow residue was dissolved in W-methyl-2-pyrrolidone (1 mL) and A/,A/-diisopropylethylamine (0.181 mL, 1.036 mmol). 2-Propenoyl ch!oride (0.034 mL, 0.415 mmol) was added and the reaction was stirred at room temperature for 1 hour. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (40 mg, 0.083 mmoi, 40 % yield) as a brown solid. LCMS (Method D): Rt 0.85 minutes; m/z 480 (MH+).
Example 87:
W-[(3R)-1 -acryloyl-3-piperidinyl]-4-(4-morphoiinylmethyl)-Af-[1 ,3]th
&]pyridin-2-yl-2,6-pyridinediamine
Figure imgf000210_0001
A microwave vial was charged with A/-[6-chloro-4-(4-morpholinylmethyl)-2- pyridinyl][1 ,3]thiazo!o[5,4-i)3pyridin-2-amine (75 mg, 0.207 mmo!), 1 , 1 -dimethylethyl (3R)- 3-amino-1 -piperidinecarboxylate (62.3 mg, 0.31 1 mmol), {1 ,3-bis[2,6-bis(1 - methylethyi)phenyl]-2-imidazolidinyl}(chloro)(2-methyl-2-propen-1 -yl)palladium (36.6 mg, 0.062 mmol). The system was sealed and placed under an atmosphere of nitrogen using a vacuum purge. Lithium bis(trimethy!siiyi)amide, 1 M in tetrahydrofuran (0.75 mL, 0.75 mmol) was added. The vial was stirred in the preheated oil bath at 70 °C for 2 hours. The vial was cooled down to room temperature and the reaction mixture was partitioned between dich!oromethane (10 mL) and saturated ammonium chloride (10 mL). After separation, the organic extract was dried using a hydrophobic frit and evaporated to dryness. The residue was dissolved in dichloromethane (3 mL) and methanol (1 mL). The solution was treated with 4M HCI in dtoxane (0.5 mL, 2 mmol) and stirred at room temperature overnight. The solvent was evaporated to dryness. The yellow residue was dissolved in A/-methyl-2-pyrrolidone (1 mL) and Λ/, V-diisopropylethylamine (0.181 mL, 1 .036 mmol). 2-Propenoyl chloride (0.034 mL, 0.415 mmol) was added and the reaction was stirred at room temperature for 1 hour. The mixture was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (35 mg, 0.073 mmol, 35 % yield) as a brown solid. LCMS (Method D): Rt 0.85 minutes; m/z 480 (MH+).
Example 88:
4-{[c s-2,6-dimethyl-4-morpholinyi]methyi}-W-[(3S)-1-(3-methyi-2-buten
pyrrolidinyl]-Ar-[1 ,3]thiazolo[5,4-b]pyridin-2-yI-2,6-pyridinediamine
Figure imgf000211_0001
4-{[c/s-2,6-Dimethy!-4-morpholiny^
jb]pyridin-2-yl-2,6-pyridinediamine dihydrochloride (75 mg, 0.146 mmol) was suspended in A/-methyl-2-pyrrolidone (0.6 mL) and stirred at room temperature. N,N- Diisopropylethylamine (0.77 mL, 0.439 mmol) was added followed by 3-methyl-2-butenoyl chloride (0.016 mL, 0.146 mmoi) and the reaction mixture was then stirred for 30 minutes. The solution was purified directly by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (60 mg, 0.1 15 mmol, 79 %). LCMS (Method E): Rt 0.70 minutes; m/z 522 (MH+).
Example 89:
W-{{3S)-1 -[(2E)-2-butenoy[]-3-pyrrolidinyl}-4-{[c s-2,6-dimethyl-4- morpholinyllmethy^-A -iljSjthiazoiotS^-blpyridin- -yl^^-pyridinediamine
Figure imgf000211_0002
4-{[c/S"2l6-Dimethyi-4-morpholinyl]methyl}-A/-E(3S)-3-pyrrolidinyi]-/\/,-[1 ,3]thiazolo[5,4- jb]pyridin-2-yl-2[6-pyridinediamine dihydrochloride (75 mg, 0.146 mmol) was suspended in A/-methyl-2"pyrrolidone (0.6 mL) and stirred at room temperature. Λ/,Λ/- Diisopropylethylamine (0.77 mL, 0.439 mmo!) was added followed by (2E)-2-butenoyl chloride (0.014 mL, 0.146 mmol) and the reaction mixture was then stirred for 30 minutes. The solution was purified by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (45 mg, 0.089 mmol, 61 %). LCMS (Method E): Rt 0.65 minutes; m/z 508 (MH+).
Example 90:
4-{[c/s-2,6-dimethyl-4-morpholinyn
pyrrolidinyi}-W-[1,3]thiazolo[5,4- j]pyridin-2-yl-2,6-pyridinediamine
Figure imgf000212_0001
Tiglic acid (39 mg, 0.39 mmol) and 1 ,1 '-carbonyldiimidazole (63 mg, 0.389 mmol) were dissolved in A/-methyl-2-pyrrolidone (0.6 mL) and stirred at room temperature for 15 minutes. -{[C s-2,6-dimethyl-4-morpholinyl]methyl}-Λ/-[(3S)-3-pyrrolίdinyl]-Λ/·- [1 >3]thiazolo[5,4-/)]pyridin-2-yi-2,6-pyridined!amine dihydrochloride (100 mg, 0.195 mmol) was suspended in W-methy!-2-pyrrolidone (0.6 mL) and stirred at room temperature. N,N- Diisopropylethylamine (0.102 mL, 0.585 mmo!) was then added and most of the solid went slowly into solution. This solution was then added to the tiglic acid and 1 ,1 '- carbonyldiimidazole mixture and the combined mixture was stirred at room temperature for a further 30 minutes. The solution was purified by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (51 mg, 0.098 mmol, 50 %). LCMS (Method D): Rt 0.99 minutes; m/z 522 (MH+).
Example 91 :
4-{[c/'s-2,6-dtmethyl-4-morpholinyl3methyl}-W-[(3S)-1-(2-methylacryloyl)-3- pyrrolidinyiJ-A -Il jSlthiazoloIS^-bl yridin^-yl^^- yridinediamine
Figure imgf000212_0002
2-Methyl-2-propenoic acid {34 mg, 0.395 mmol) and 1 ,1 '-carbony!diimidazole (63 mg, 0.389 mmol) were dissolved in /V-methyl-2-pyrrolidone (0.6 mL) and stirred at room temperature for 15 minutes. 4-{[C/s-2,6-dimethy[-4-morpholinyl]methyi}»A/-[(3S)-3- pyrrolidtny1]-A/,-[1 ,3]thiazolo[5,4- j]pyr!din-2-yl-2,6-pyridinediamine dihydrochioride (100 mg, 0.195 mmol) was suspended in A/-methyl-2-pyrrolidone (0.6 mL) and stirred at room temperature, Λ/,/V-Diisopropyiethylamine (0.102 mL, 0.585 mmoi) was then added and most of the solid went slowly into solution. This solution was then added to the 2-methyl- 2-propenoic acid and 1 ,1 '-carbonyldiimidazole mixture and the combined mixture was stirred at room temperature for a further 30 minutes. The solution was purified by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (46 mg, 0.09 mmol, 46 %). LC S (Method D): Rt 0.95 minutes; m/z 508 (MH+).
Example 92:
N-{{3S)-1 -E(2£)-4-(dimethylamino)-2-butenoy^
morpholinyl]methyl}-AT-[1 ,3]thiazolo[5,4-) ]pyridin-2-yl-2,6-pyridinediamin
Figure imgf000213_0001
A round bottom flask was charged with 1 , 1-dimethylethyl (3S)-3-{[4-{[c/s-2,6-dimethyi-4- morpho!inyl]methyl}-6-([1 ,3]thiazolo[5,4-/b]pyridin-2-ylamino)-2-pyridinyl]amino}-1- pyrrolidinecarboxylate (104 mg, 0.193 mmol), 4M HCI in dioxane (1.893 mL, 7.57 mmo!), dichloromethane (3 mL) and methanol (2 mL). The reaction was stirred at room temperature for 1 hour. The solvent was removed under reduced pressure. The residue was dissolved in A/-methyl-2-pyrroiidone (2 mL) and treated with Λ/,Λ/'- diisopropylethylamine (0.101 mL, 0.578 mmol). This solution was added to a stirred mixture of 2-(7-aza-1 H-benzotriazo!e-1 -yi)-1 ,1 ,3,3-tetramethyluronium hexafluorophosphate (110 mg, 0.289 mmol), (2£)-4-(dimethylamino)-2-butenoic acid hydrochloride (47.9 mg, 0.289 mmol) and /V,A/-diisopropyiethylamine (0.101 mL, 0.578 mmol) in W-methyl-2-pyrrolidone (2 mL). The reaction was stirred at room temperature for 1 hour. The mixture was purified by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (47 mg, 0.085 mmol, 44 %) as an off-white soiid. LCMS (Method D): Rt 0.89 minutes; m/z 551 (MH+).
Example 93:
4-{[c/s-2(6-dimethyl-4-morpholinyl]methyi}-W-{{3S)-1 -[2-(4- morpholinylmethyl)aciyloyl]-3^yrrolidinyl}-AT-[1 ,3]thiazolot5,4-Jb3pyridin-2-yl-2,6-
Figure imgf000214_0001
2-(4-Morpholinylmethyl)-2-propenoic acid (42.8 mg, 0.250 mmol) was suspended in N- methyl-2-pyrrolidone (0.5 mL). 2-(7-Aza-1 H-benzotriazole-1 -yl)-1 , 1 ,3,3- tetramethyluronium hexafluorophosphate (130 mg, 0.341 mmol) was added and the reaction was stirred at room temperature for 30 minutes. A solution of 4-{[c s-2,6-dimethyl- 4-morpho!inyl]methyl}-A/-[(3S)-3-pyrrolidinyl]-/\/,-[1 ,3]thiazolo[5,4-ib]pyridin-2-yl--2l6- pyridinediamsne (100 mg, 0.227 mmol) and Λ/,/V-diisopropylethylamine (0.1 19 mL, 0,682 mmol) in W-methyl-2-pyrrolidone (0.5 mL) was added and the reaction was stirred at room temperature for 3 hours. The reaction was stirred at room temperature for 1 hour. The mixture was purified by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (73 mg, 0.123 mmol, 54 %). LCMS (Method D): Rt 0.91 minutes; m/z 593 (MH+).
Example 94:
W-(1-acryioyi-3-azetidinyl)-4-(4-morpholinylm^
yi-2,6-pyridinediamine
Figure imgf000215_0001
A round bottom flask was charged with 1 , 1 -dimethylethy I 3-{[4-(4-morpholinyimethyl)-6- ([1 ,3]thiazolo[5,4-b]pyridin-2-ylamino)-2-pyridinyl]amino}-1-azetidinecarboxylate (53 mg, 0.107 mmol), 4M HC! in dioxane (1.065 mL, 4.26 mmol), dichloromethane (3 mL) and methanol (2 mL). The resultant reaction mixture was stirred at room temperature for 1 hour. A second portion of 4M HCI in dioxane (1 .206 mL, 4.82 mmol) was then added and the reaction was stirred at room temperature for a further 1 hour. The solvent was removed under reduced pressure. The residue was dissolved in A/-methyl-2-pyrrolidone (1 mL) and treated at 0 °C with A/.W-diisopropylethylamine (0.093 mL, 0.533 mmol) followed by 2-propenoyl chloride (0.009 mL, 0.107 mmol). The reaction was then stirred at 0 °C for 1 hour. The mixture was purified directly by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (27 mg, 0.06 mmol, 56 %) as a brown solid. LCMS (Method D). Rt 0.80 minutes; m/z 452 (MH+). Example 95:
W-[(3S)-1-acryloyl-3-pyrroiidinyl]-4-^
i) ridin-2-yl-2,6-pyridinediamine
Figure imgf000215_0002
A round bottom flask was charged with 1 , 1 -dimethylethyl (3S)-3-{[4- [(dimethylamino)methyl]-6-([1 ,3]thiazolo[5,4-0] pyridine-2-ylamino)-2-pyridinyl]amino}-1 - pyrrolidinecarboxylate (20 mg, 0.043 mmol), 4 HCI in dioxane (0.426 mL, 1.704 mmol), dichloromethane (1 mL) and methanol (0.667 mL). The reaction was stirred at room temperature for 1 hour. A further portion of 4M HCI in dioxane (0.426 mL, 1.704 mmol) was then added and and stirring continued at room temperature for 1 hour. The solvent was removed under reduced pressure. The residue was dissolved in /V-methyl-2- pyrrolidone (1 ml_) and treated at 0 °C with A ,A/-diisopropyiethylamine (0.037 mL, 0.213 mmol) and 2-propenoyl chloride (0.003 mL, 0.043 mmol). The reaction was stirred at 0 °C for 1 hour. The mixture was purified directly by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (7.7 mg, 0.018 mmol, 37 %). LCMS (Method D): Rt 0.84 minutes; m/z 424 (MH+).
Example 96:
W-[(3R)-1-acryloy(-3-pyrrolidinyl]-^^
trimethylpropyl3amino}rnethyi)-2J6-pyridinediamine
Figure imgf000216_0001
A round bottom flask was charged with 1 , 1-dimethylethyl (SRJ-S- -ill .Sjthiazolo^^- jb]pyridin-2-yiamino)-4-({[(1 S)-1 ,2,2-trimethylpropyl3amino}methyl)-2-pyridinyl]amino}-1- pyrrolidinecarboxylate (85 mg, 0.162 mmol), 4M HCI in dioxane (1.617 mL, 6.47 mmol), dichloromethane (2 mL) and methanol (1.33 mL). The reaction was stirred at room temperature for 1 hour. A further quantity of 4M HCI in dioxane (1.617 mL, 6.47 mmoi) was added and the reaction was stirred at room temperature for 2 further hours. The solvent was then removed under reduced pressure. The residue was dissolved in N- methyl-2-pyrrolidone (1 mL) and treated at 0 °C with W,/V-diisopropylethylamine (0.141 mL, 0.808 mmol) and 2-propenoyl chloride (0.013 mL, 0.162 mmol) added dropwise. The reaction was stirred at 0 °C for 1 hour. The mixture was purified directly by mass directed automated preparative HPLC (trifluoroacetic acid modifier) to afford, after basification with aqueous ammonia, the title compound (43 mg, 0.09 mmol, 55 %) as a white solid. LCMS (Method E): Rt 0.68 minutes; m/z 480 (MH+). Example 97:
W^KSSJ-l-acryloyl-a-pyrrolidinyll-^-te-ethyl-l ^-benzothiazol^-ylJ-e- (phenylmethyl)-2,4-pyrimidinediamine
Figure imgf000217_0001
A microwave vial was charged with 6-ethyl-A/-[2-fluoro-6-(phenyimethyl)-4-pyrimidinyi]- 1 ,3-benzothiazoi-2-amine (which may be prepared as described in WO 2010/106016) (45 mg, 0.123 mmol), 1 ,1 -dimethylethyl (3S)-3-amino-1-pyrrolidinecarboxylate (69.0 mg, 0.370 mmol) and isopropanol (3 mL). The vial was sealed and heated under microwave irradiation for 1 hour at 130 °C. The solvent was removed under reduced pressure. The mixture was purified directly by mass directed automated preparative HPLC (formic acid modifier gradient) to afford the required intermediate. The solid was dissolved in methanol (1 mL) and dichloromethane (2 mL) and treated with 4 HCI in dioxane (0.309 mL, 1.235 mmol). The reaction was stirred at room temperature for 16 hours. The solvent was removed under reduced pressure. The residue was dissolved in /V-methyl-2-pyrrolidone (0.8 mL) and treated with A ,W-diisopropylethyiamine (0.065 mL, 0.370 mmol) and 2- propenoyl chloride (0.005 mL, 0.062 mmoi). The reaction was left standing at room temperature for 30 minutes and was then purified directly by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (20 mg, 0.041 mmol, 33 %). LCMS (Method A): Rt 0.95 minutes; m/z 485 (MH+).
Example 98:
W-[(3S)-1-acryloyl-3-pyrrolidinyl]-W1-1 ,3-benzothiazol-2-yl-4-(4-morpholinyl methyl)- 2,6-pyridinediamine
Figure imgf000217_0002
A round bottom f!ask was charged with 1 , 1-dimethylethyi (3S)-3-{[6-(1 s3-benzothiazol-2- ylamino)-4-(4-morpholinylmethyl)-2-pyridiny!]amino}-1 -pyrrolidine carboxylate (160 mg, 0.313 mmol) , dichloromethane (2 mL) and methanol (2 mL). HCI in dioxane (4M) (1 mL, 4.0 mmol) was added and the reaction was stirred for 16 hours. The solvent was removed under reduced pressure. The residue was dissolved in A/-methyl-2-pyrrolidone (1 mL) and Λ/,/V-diisopropylethylamine (0.274 mL, 1 .567 mmol). 2-Propenoyl chloride (0.025 mL, 0.313 mmol) was added and the reaction stirred for 30 minutes and was then purified directly by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (67 mg, 0.144 mmoi, 46 %). LC S (Method D): Rt 0.94minutes; m/z 465 (MH+). Example 99:
W-KSSJ-l-acryioyl-S-pyrrolidinyll-AT-te-imethyloxyJ-l^-benzothiazol^-yl]^^- morpho!inylmethyi)-2,6-pyridinediamine
Figure imgf000218_0001
A round bottom flask was charged with 1 ,1-dimethyiethyl (3S)-3-{[6-{[6-(methyloxy)-1 ,3- benzothiazol-2-yl]amino}-4-(4-morpholiny!methyl)-2-pyridinyl]amino}-1 - pyrrolidinecarboxylate (85 mg, 0.157 mmol), dichloromethane (2 mL) and methanol (2 mL). HCI in dioxane (4M) (1 mL, 4.0 mmol) was added and the reaction was stirred for 16 hours. The solvent was removed under reduced pressure. The residue was dissolved in V-methyl-2-pyrrolidone (1 mL) and /V,A/-diisopropylethylamine (0.137 mL, 0.786 mmol). 2- Propenoyl chloride (0.013 mL, 0.157 mmol) was added and the reaction stirred for 30 minutes. The residue was purified by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (37 mg, 0.075 mmol, 48 %). LCMS (Method D): Rt 0.93minutes; m/z 495 (MH+). Example 100:
W-[(3S)-1 -acryloy!-3-pyrrolidiny Ι]-ΛΓ-1 ,3-benzothiazol-2-yi-4-({[(1 S)-1 ,2,2- trimethy!propyl]amino}methyl)-2,6-pyridinediamine
Figure imgf000219_0001
A round bottom flask was charged with 1 , 1-dimethylethyl (3S)-3-{[6-(1 ,3-benzothiazol-2- ylamino)-4-({[(1 S)-1 ,2,2-trimethylpropyl]amino}methyl)-2-pyridinyl]amino}-1- pyrro!idinecarboxylate (37 mg, 0.071 mmol) and dichloromethane (5 mL). 4M HCI in dioxane (0.5 mL, 2.0 mmol) was added and the reaction was stirred at room temperature for 16 hours. The solvent was removed under reduced pressure. The residue was dissolved in A/-methyl-2-pyrrolidone (1 mL) and W,/V-diisopropylethylamine (0.062 mL, 0.355 mmol). 2-Propenoyl chloride (0.006 mL, 0.070 mmol) was added and the reaction stirred for 10 minutes. The residue was purified by mass directed automated preparative HPLC (trifluoroacetic acid modifier) to afford, after basification, the title compound (14 mg, 0.029 mmol, 41 %) as a yellow solid. LCMS (Method E); Rt 0.77 minutes; m/z 479 (MH+).
Example 101 :
W-[(3S)-1-acryloyl-3-pyrrolidinyl3-/\T-[6-(methyIoxy)-1 ,3-benzothiazol-2-yl3-4-({K 1 -trimethylpropyl]amino}methyl)-2,6-pyridinediamine
Figure imgf000219_0002
A round bottom flask was charged with 1 ,1 -dimethylethyl (3S)-3-{[6-{[6-(methyloxy)-1 ,3- benzothiazoi-2-yl]amino}-4-({[(1 S)-1 ,2,2-trimethylpropyl]amino}methy!)-2-pyridinyi]amino}- 1-pyrrolidinecarboxylate (37 mg, 0.067 mmol) and was dissolved in dichloromethane (5 mL). 4M HCI in dioxane (0.5 mL, 2.0 mmol) was added and the reaction was stirred at room temperature for 16 hours. The solvent was removed under reduced pressure. The residue was dissolved in A/-methyi-2-pyrrolidone (1 mL) and A/,A/-diisopropyiethy!amine (0.058 mL, 0.333 mmol). 2-Propenoyl chloride (0.006 mL, 0.070 mmol) was added and the reaction stirred for 10 minutes. The residue was purified by mass directed automated preparative HPLC (trifluoroacetic acid modifier) to afford, after basification, the title compound (12 mg, 0.023 mmol, 35 %) as a brown solid. LCMS (Method D): Rt 1.19 minutes; m/z 509 (MH+).
Example 102:
W-[{3R)-1 -acryloyl-3-pyrrolidinyl]-/^
imethylpropyl]amino}methyl)-2,6-pyridinediamine
Figure imgf000220_0001
A round bottom flask was charged with 1 , 1-dimethylethyl (3 )-3-{[6-([1 ,3]thiazo!o[5,4-
Figure imgf000220_0002
pyrrolidine carboxylate (262 mg, 0.498 mmol), 4M HCI in dioxane (4.98 mL, 19.93 mmol), dichloromethane (2 mL) and methanol (1.33 mL). The reaction was stirred at room temperature for 1 hour. A further quantity of 4M HCi in dioxane (4.98 mL, 19.93 mmol) was added and the reaction was stirred at room temperature for 2 further hours. The solvent was removed under reduced pressure. The residue was dissolved in A/-methyl~2- pyrrolidone (1 mL) and treated at 0 °C with A/,A/-diisopropylethylamine (0.435 mL, 2.492 mmoi) and 2-propenoyl chloride (0.040 mL, 0.498 mmol) added dropwise. The reaction was stirred at 0 °C for 1 hour. The solution was purified directly by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (76 mg, 0.158 mmoi, 32 %). LCMS (Method D): Rt 1.08 minutes; m/z 480 (MH+).
Example 103:
W-[(3S)-1 -acryloyl-3-pyrrolidinyl]-4-{[c s-2,6-dimethyl-4-morpholinyl]methyl}
(methyloxy)[1 ,3]thiazoio[5,4-i)]pyridin-2-yl]-2,6-pyridinediamine
Figure imgf000221_0001
A/-(6-Chloro-4-{[c/s-2,6-dimethyl-4-morpholinyl]methyl}-2-pyrtdin
(methyloxy)[1 ,3]thiazolo[5,4-Jb]pyridin-2-amine (67 mg, 0.160 mmol) and 1 ,1 -dimethylethyi (3S)-3-amino-1-pyrrolidinecarboxylate (0.031 mL, 0.176 mmol) were dissolved in tetrahydrofuran (1 mL) and heated to reflux. {1 ,3~Bis[2,6-bis(1-methylethyl)phenyl]-2- imidazolidinyl}(chloro)(2-methyl-2-propen-1 -yl)palladium (28.2 mg, 0.048 mmol) dissolved in lithium bis(trimethylsilyl)amide, 1 M in tetrahydrofuran (0.5 mL, 0.5 mmol) was added in three portions over 15 minutes and the reaction heated at reflux for 30 minutes. Further {1 ,3-Bis[2,6-bis(1-methylethyi)phenyl]-2-imidazoiidinyl}(chloro)(2-methyl-2-propen-1 - yl)palladium (28.2 mg, 0.048 mmol) dissolved in lithium bis(trimethylsilyl)amide, M in tetrahydrofuran (0.5 mL, 0.05 mmol) was added in two portions over 10 minutes and the reaction heated to reflux for 15 further minutes. The reaction was allowed to cool to room temperature and water (3 mL) and 2-methyltetrahydrofuran (5 mL) added. The layers were separated and the aqueous layer extracted with 2-methyltetrahydrofuran (2 x 5 mL). The combined organic layers were dried with anhydrous magnesium sulfate. Hydrochloric acid (5-6 M in isopropanol, 1 mL) was added and the solvent evaporated under reduced pressure to give a red gum. The gum was dissolved in methanol (2.5 mL) and the slurry was stirred for 30 minutes before being filtered. The solid was washed with methanol (2 x 5 mL), The solid and the filtrate were recombined. The solvent was evaporated under reduced pressure to give a red gum. Methanol (2.5 mL) was added and the slurry was filtered. The solid was washed with methanol (2 x 2.5 mL). The filtrate was evaporated under reduced pressure to give a red gum which was suspended in dichloromethane (1 mL). A/./V-Diisopropylethylamine (0.279 mL, 1.596 mmol) was added followed by 2- propenoic acid (0.009 mL, 0.128 mmol). The reaction was cooled to 0 °C and propylphosphonic anhydride (50% w/w solution in ethyl acetate, 0.065 mL) added. The reaction was stirred for 1 hour and water (2 mL) and dichloromethane (5 mL) were added, The layers were separated and the aqueous layer extracted with dichloromethane (2 x 5 mL), The combined dichloromethane layers were evaporated under reduced pressure. The residue was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (8.5 mg, 0.016 mmol, 10 % yield) as a yellow solid. LCMS (Method D): Rt 1.00 minutes; m/z 524 (MH+).
Example 104:
W-[(3S)-1-acr7foyi-3-pyrroIidinyl]-Ar-^
( 1 S)-1 ,2J2-trimethylpropyl]amino}methyl)-2)6-pyridinediamine
Figure imgf000222_0001
A/-t6-Chloro-4-({[(1 S)-1 ,2,2-trimethylpropyl]amino}methyl)-2-pyridinyl]-5- (methyloxy)[1 ,3]thiazolo[5,4-6]pyridin-2-amine (50 mg, 0.123 mmol) was weighed into a microwave reactor vial and 1 ,1 -dimethylethyl (3S)-3-amino-1-pyrrolidinecarboxylate (0.024 ml, 0.135 mmol) dissolved in anhydrous tetrahydrofuran (1 mL) was added. The system was sealed and placed under an atmosphere of nitrogen using a vacuum purge and heated to reflux. {1 ,3-Bis[2,6-bis(1-methyiethyl)phenyl]-2-imfdazo[idiny!}(chloro)(2- methyl-2-propen-1-yl)palladium (21.8 mg, 0.037 mmol) dissolved in lithium bis(trimethylsilyl)amide, 1 M in tetrahydrofuran (0.37 mL, 0.37 mmol) was added in three equal portions over 15 mintues. The reaction was heated to reflux for 30 minutes. Water (2.5 mL) was added and the reaction cooled to room temperature. 2- Methyltetrahydrofuran (5 mL) was added and the layers were separated. The aqueous layer was extracted with 2-methyltetrahydrofuran (2 x 5 mL), the combined layers were dried using a hydrophobic frit and the solvent evaporated under reduced pressure to give a brown sludge. Hydrochloric acid (5-6 M in isopropanoi, 3 mL) was added followed by methanol (3 mL) and the reaction stirred for 1 hour. The reaction solvent was evaporated under reduced pressure to give a brown solid. Anyhydrous dichloromethane (3 mL) was added followed by W, V-diisopropy!ethylamine (0.43 mL, 2.463 mmol) and 2-propenoic acid (0.006 mL, 0.086 mmol). The solution was cooled to 0 °C and propylphosphonic anhydride (50% w/w solution in ethyl acetate, 0.05 mL, 0.084 mmo!) was added. The reaction was stirred for 20 minutes. The reaction was warmed to room temperature and water (3 mL) was added. The layers were separated and the aqueous layer extracted with dichloromethane (3 mL). The combined dichloromethane layers were dried using a hydrophobic frit and evaporated under reduced pressure to give a brown solid. Dimethyl sulfoxide (1.750 mL), methanol (2 mL) and hydrochloric acid (5-6 M in isopropanol, 2 mL) were added to the solid. The slurry was filtered to give a clear solution. The solution was evaporated under reduced pressure to give a concentrated brown solution (3 mL) which was subjected to purification by mass directed automated preparative HPLC (formic acid modifier) to afford a brown solid (19 mg). This solid was subjected once more to purification by mass directed automated preparative HPLC (formic acid modifier) to afford, after basification with sodium bicarbonate, the title compound (2 mg, 0.004 mmo!, 3 % yield) as a yellow solid. LCMS (Method A): Rt 0.77 minutes; m/z 510 (MH+). Example 105:
W (3S)-1-acryioyl-3-pyrrolidiny[3-4-[(dimethylamino)methyl]-W'-[5- methyioxy)[1J3]thiazolo[5,4-ft]pyridin-2-yl]-2,6-pyridinediamine
Figure imgf000223_0001
1 ,1 -Dimethylethyl (3S)-3-[(4-[(dimethylamtno)methyl]-6-{[5-(methy!oxy)[1 ,3]thiazolo[5,4- i¾3pyndin-2-yl]amino}-2-pyridiny!)amino]-1 -pyrrolidinecarboxylate (50 mg, 0.100 mmol) was dissolved in methanol (0.6 mL), hydrochloric acid in isopropanol (5-6 M, 0.04 mL, 0.2 mmol) added and the mixture stirred for 3 hours. The solvent was evaporated under reduced pressure. The resulting solid was dissolved in MeOH (3 mL) and the solvent was evaporated under reduced pressure to give a solid. The solid was suspended in dichioromethane (1 mL). W,A/-Diisopropylethylamine (0.087 mL, 0.500 mmol) was added and the solution was cooled to 0 °C. 2-Propenoic acid (0.007 mL, 0.105 mmol) and propylphosphonic anhydride (50% w/w solution in ethyl acetate, 0.060 mL, 0.1 mmol) were added. The reaction was stirred for 15 minutes at 0 °C, allowed to warm to room temperature and stirred for 2 hours. Water (2 mL) and dichioromethane (2 mL) were added, the layers separated and the aqueous layer extracted with dichioromethane (2 5 mL). The combined dichioromethane layers were dried using a hydrophobic frit and evaporated under reduced pressure. The residue was subjected to purification by mass directed automated preparative HPLC (ammonium bicarbonate modifier) to afford the title compound (2 mg, 0.004 mmol, 4 % yield) as a yellow solid. LCMS (Method A): Rt 0.72 minutes; m/z 454 (MH+). Additional compounds which may be prepared by similar methods include:
Example 106:
W-[{3S)-1-acryloyl-3-pyrrolidmyl]-4-[(dim^
benzothi zol-2-yl]-2,6-pyridinediamine
Figure imgf000224_0001
Example 107:
W-[(3S)-1 -acryloyl-3-pyrrolidinyl]-/V-1 ,3-benzothiazol-2-y I-4- dimethylamino)methyl]-2,6-pyridinediamine
Figure imgf000224_0002
SALT PREPARATION W-[(3S)-1 -Acryloyl-3-pyrrolidinyl]-W-[1 ,3Ithiazolo[5,4-i>]pyndin-2-yl-4-({[(1 S)-1 ,2,2- trimethylpropyl]amino}methyl)-2,6-pyridinediamine difumarate monohydrate
Method A
Fumaric acid (6.35 mg, 0.055 mmol, 1.05 eq) was dissolved in tetrahydrofuran and added to ^-[(SSJ-l-acryloyl-S-pyrrolidinylj-W-ttSjthiazolotS^-^pyridin^-yl^-iiKI SJ-l ^^- trimethylpropyl]amino}methyi)-2,6-pyridinediamine (25 mg, 0.052 mmo!). This was sonicated and temperature cycled from 0-40 °C for 4 days. After this time, some solids were seen to be birefringent by polarised light microscopy (PLM) so ca. 0.1 mi solvent was removed by evaporation under a nitrogen stream to give more solids, however these appeared amorphous by PLM. This suspension was allowed to temperature cycle overnight. The solids still appeared amorphous, so the solvent was evaporated under a nitrogen stream to give an oil/gum. This was treated with water (0.05 mL) which, on heating and sonication, dissolved the gum. The solution was allowed to cool and showed no evidence of precipitation so further water (1 mL) was added which gave a hazy solution. This hazy solution was allowed to temp cycle overnight before being filtered and sucked free from solvent.
Method B
Fumaric acid (225.2 mg, 1.94 mmol, 2.04 eq) was dissolved in tetrahydrofuran (1.82 mL) and water (0.9m!) with heating and sonication. This acidic solution was added to N-[(3S)~ 1 -acryloyl-3-pyrrolidinyl]-W-[1 ,3]thiazolo[5,4- ]pyridin-2-yl-4-({[(1 S)-1 ,2,2- trimethylpropyl]amino}methyi)-2,6-pyridinediamine (455.4 mg, 0.95 mmol), which was gently warmed and sonicated to give a clear orange solution. This was treated with water (4.51 mL) and then seeded with previously prepared difumarate monohydrate salt (~ 1 mg). This suspension was allowed to stir for 10 minutes to allow the crystallisation to initiate before the remaining water (4.51 mL) was added. The suspension was allowed to stir at room temperature overnight. After this period, the particles were still very small so the suspension was temperature cycled 0-40 °C for 3 days. The suspension was filtered and sucked free from solvent before drying further in vacuo at 53 °C to give V-[(3S)-1- acryloyl-3-pyrrolidinyl]-W-[1 ,3jthiazolo[5,4-6]pyridin-2-yl-4-({[(1 S)-1 ,2,2- trimethylpropyl]amino}methyl)-2,6-pyridinediamine difumarate (518.7 mg, 75%th)
NMR: consistent with difumarate salt formation
NMR (400MHz, d6-DMSO): d = 11.4 (bs, 0.7H), 8.27 (d, J = 4.7Hz, 1 H), 7.86 (d, J = 7.8Hz, 1 H), 7.37 (dd, J = 4,7 and 8Hz, 1 H), 7.05 (dd, J = 6.6 and 15.2Hz, 1 H), 6.61 (m, 5H), 6.34 (s, 1 H), 6.23 (s, 1 H), 6.17 (m, 1 H), 5.70 (m, 1 H), 4.70 (m, 1 H), 4.00 (m, 0.5H), 3.84 (d, J = 14.4Hz, 1 H), 3.73 (m, 1.6H), 3.66 (m, 3H), 2.38 (m, 1.5H), 2.25 (m, 0.5H), 2.04 (m, 1 H), 1.02 (d, J = 5.1 Hz, 3H) and 0.9 (s, 9H)
Fractional integrals are due to rotomers which are resolved at ambient temperature.
Thermogravametric analysis (TGA) shows 2.5%w/w loss in mass on heating to 142°C which corresponds to 1eq water.
POLYMORPH EXPERIMENTAL
N-[(3S)-1 -Acryloyl-3-pyrrolidinyl]-^
trimethylpropyi]amino}methyi)-2,6-pyridinediamine difumarate monohydrate
X-Ray Powder Diffraction (XRPD) The data were acquired on a PANa!yiical X'Pert Pro powder diffractometer; model PW3040/60 using an X'Ceierator detector. The acquisition conditions were: radiation: Cu Ka, generator tension: 40 kV, generator current: 45 mA, start angle: 2.0° 2Θ, end angle: 40.0° 2Θ, step size: 0.0167° 2Θ, time per step: 31.75 seconds. The sample was prepared by mounting a few milligrams of sample on a silicon wafer (zero background) plate, resulting in a thin layer of powder.
The XRPD data are shown in Figure 1. Characteristic XRPD angles and d-spacings are recorded in Table 1 below. Peak positions were measured using Highscore software.
Figure imgf000226_0001
Table 1
BIOLOGICAL DATA
1. Itk Homogeneous Time Resolved Fluorescence (HTRF)
The activity of recombinant human Itk (full length) is assessed using an HTRF assay with truncated human SAM-68 (R331 -Y443) as the substrate.
Recombinant human Itk (full length) is expressed in insect cells (in pFastBac-1 vector Invitrogen) fused to a Flag tag at its N terminus. The sequence of the Itk part is identical to Genbank entry L10717. The FLAG-ltk fusion protein is extracted from insect cells and purified by immunoaffinity chromatography on anti-FLAG (M2) agarose affinity resin. Further purification is by size exclusion chromatography. Purified protein is stored at - 80°C in Tris-HCI (50m ), NaCI (200mM), sorbitol (500mM), DTT (2mM), pH 8.0.
Truncated human SAM-68 (R331-Y443) is expressed in E. coli (using a pGex-4T vector Pharmacia) as a GST-thrombin cleavage site-Avi-tag-Sam68:331-443 fusion. The Sam68 part of the fusion (R331-Y443) is identical to the sequence of Genbank database entry NM__006559. GST-SAM68 is purified by affinity chromatography on glutathione- sepharose. Specific biotinylation of the Avitag sequence of GST-SAM68 is performed at room temperature in the presence ofmg:ATP, (5mM), D-biotin, (1 mM), DTT, (1 m ) and biotin ligase, (1 uM), and is complete in 2 hours. The biotinylated protein is further purified by size exclusion chromatography and stored at -80°C in Tris-HCI (50m ), NaCI (250mM), glycerol (10%), DTT (2mM), pH 8.0. itk (typically 5-50μΜ) is pre-activated by incubation with 100μΜ ATP and 10mMmgCI2 for 30 minutes at room temperature before dilution in assay buffer (50mM HEPES, 1 mM dithiothreitol, 0.0025% Tween-20, pH7.4) to give a concentration which ensures linearity proportional to time and enzyme concentration (typically a 5nM final concentration in the assay). Compounds at various concentrations (typical range 25pM - 25μΜ) or DMSO vehicle (at less than 5% final assay concentration) are incubated with 0.003 mL substrate (final assay concentrations 50nM biotinylated GST SAM68, 10mMmgCI2, 20μΜ ATP in 50mM HEPES, 1 mM DTT, 0.0025% Tween 20, pH7.4). The activated Itk enzyme (0.003 mL volume,) is added to initate the phosphorylation reaction. Following an incubation at 20°C, (for a time determined to ensure the assay remains in linear initital rate phase, typically 30 minutes), the reaction is halted by adding stop/read reagent (0.003 mL). The stop/read reagent comprises streptavidin APC (50nM final assay concentration; Perkin Eimer), europium-anti-phosphotyrosine antibody (0.5nM final assay concentration; Wallac) diluted in 40mM HEPES, 150mM NaCI, 0.03%w/v BSA, 60mM EDTA. The assay plates are left to equilibrate for at least 45 minutes at 20°C, before reading on a suitable HTRF reader.
The compounds of Examples 1 to 102 were tested in the above or a similar assay and were found to have a mean pKi of 5 or greater.
2. Assessment of the potential for irreversible inhibition of itk
Method A
Method 1A: Assessing initial rates of binding of compounds to Itk using the Invitrogen LanthScreen kinase binding assay
General principle:
The assay is based on the binding and displacement of a proprietary, A!exa-fluor 647- labelled ATP-competitive kinase inhibitor tracer. Binding of the tracer is detected using a europium-labelled anti-tag antibody which binds to the kinase of interest. Simultaneous binding of the tracer and antibody results in FRET; displacement of the tracer upon inhibitor binding results in a decrease of FRET. The assay can be read continuously, enabling rates of binding (and displacement) to be monitored.
Assay components were as follows:
Kinase tracer 236 (Invitrogen PV5592): The binding of the tracer 236 is reversible, rapid (to.5 for association - 27s) with a Kd of 36nM.
Lanthscreen Eu-anti-GST antibody (Invitrogen PV5594)
GST-tagged recombinant ITK (Invitrogen PV3875)
kinase buffer A (Invitrogen PV3189): 50mM HEPES pH7.5, 0.01 % (w/v) Brij-35, 10m MgCI2, 1 mM EGTA A 16-point (1 :3 dilution) compound titration was prepared in kinase buffer A containing 1 % (v/v) DMSO, starting at 300 μΜ (3x FAC (final assay concentration)), and including a zero- compound 1 % DMSO control. A 3x FAC pre-mix of 15nM GST- ITK and 15nM Eu-anti- GST antibody, diluted in kinase buffer A, was allowed to complex for 20minutes at room temperature, prior to addition of 5μΙ to 5μΙ compound in a 384-well Greiner low-volume plate, along with 5μΙ of 90nM (3x FAC) kinase tracer 236, diluted in kinase buffer A. The plate was read immediately on an Envision plate reader (laser excitation, dual emission filters 665nm and 615nm); data were collected over a period of 4hours at 2 minute intervals and the ratio of the 665nm:615nm emission counts, calculated. The rate of compound binding is coincidental with the apparent decrease in the FRET signal.
Data were fitted to equation 1) for each compound concentration. The resulting kobs was then plotted against compound concentration, generating a hyperbolic profile consistent with 2-step, irreversible binding.
Eqn 1 A = A0 + /l1e_feoi'ii where A = signal change, A0 = signal at time infinity, = signal at time t
The data were fitted to equation 2) to generate K, and kinact (inhibitor affinity driving the initial non-covalent interaction, and maximal rate of irreversible inactivation, respectively). The ratio kinact I Kt was then used as a measure of comparison.
Eqn 2 kobs - where kinact = maximal rate of irreversible inactivation, [I] =
Figure imgf000229_0001
concentration of compound, [L] = concentration of tracer 236, Kd = affinity of tracer236 for ITK, Ki = inhibitor affinity driving the initial non-covalent interaction. Method 2A: Assessing the reversibility of the compound : ITK complex through rapid dilution into an excess of competing ligand {"jump-dilution")
This method is necessary to complement the data generated in method 1A, since the hyperbolic profiles generated by fitting kobs vs inhibitor concentration could also be consistent with slow-binding reversible inhibition. The protocol is essentially as for method 1A, with the following modifications:
A 3x FAC pre-mix of 15nM GST-ITK and 15nM Eu-anti-GST antibody, diluted in kinase buffer A, was allowed to complex for 20minutes at room temperature, prior to addition of 5 μΙ to 5μ! compound in a 384-well Greiner low-volume plate. The interaction between GST- ITK-Ab pre-mix and the varying concentrations of compound was allowed to complex for 2hours, prior to the addition of 5μΙ 900 nM (3x FAC) of kinase tracer 236. The plate was read immediately on an Envision plate reader (laser excitation, dual emission filters 665nm and 615nm); data were collected over a period of 6 hours at 2 minutes intervals, and the ratio of the 665nm:615nm emission counts was calculated. The rate of compound displacement is coincidental with the apparent increase in the FRET signal (since the rapid binding of the tracer is not the rate-limiting step). Where no apparent displacement was observed, the IC50 was calculated and compared to that predicted by equation 3), taking into account the higher (10 x Kd) concentration of the competing tracer. The ratio of the predicted IC50 vs the measured IC50 was used to guide whether the data was consistent with the compound acting as an irreversible inhibitor (ie, the measured /C50was significantly more potent than predicted under these conditions).
Figure imgf000230_0001
Method B
Method 1 B: Assessing initial rates of binding of compounds to Itk using the Invitrogen LanthScreen kinase binding assay
General principle:
The assay is based on the binding and displacement of a proprietary, A!exa-fluor 647- labelled ATP-competitive kinase inhibitor tracer. Binding of the tracer is detected using a europium-labelled anti-tag antibody which binds to the kinase of interest. Simultaneous binding of the tracer and antibody results in FRET; displacement of the tracer upon inhibitor binding results in a decrease of FRET. The assay can be read continuously, enabling rates of binding (and displacement) to be monitored.
Assay components were as follows:
Kinase tracer 236 (Invitrogen PV5592): The binding of the tracer 236 is reversible, rapid (to.5 for association ~ 27 s) with a Kd of 36 nM.
Lanthscreen Eu-anti-GST antibody (Invitrogen PV5594)
GST-tagged recombinant ITK (Invitrogen PV3875)
kinase buffer A (Invitrogen PV3189): 50 mM HEPES pH7.5, 0.01 % (w/v) Brij-35, 10 mM MgCI2, 1 mM EGTA
A 16-point (1 :3 dilution) compound titration was prepared in kinase buffer A containing 1 % (v/v) DMSO, starting at 300 μΜ (or 100 μΜ) (3x FAC (final assay concentration)), and including a zero- compound 1 % DMSO control. A 3x FAC pre-mix of 15 nM GST-!TK and 15 nM Eu-anti-GST antibody, diluted in kinase buffer A, was allowed to complex for 20minutes at room temperature, prior to addition of 5 μΙ to 5 μ! compound in a 384-well Greiner low-volume plate, along with 5 μΙ of 90 nM (3x FAC) kinase tracer 236, diluted in kinase buffer A. The plate was read immediately on an Envision plate reader (laser excitation, dual emission filters 665 nm and 615 nm); data were collected over a period of 4 hours at 2 minute intervals (or 5 hours at 300 second intervals) and the ratio of the 665 nm:615 nm emission counts, calculated. The rate of compound binding is coincidental with the apparent decrease in the FRET signal.
Data were fitted to equation 1 ) for each compound concentration. The resulting kobs was then plotted against compound concentration, generating a hyperbolic profile consistent with 2-step, irreversible binding. Eqn 1 A = A0 + A1e~k°bst where A = signal change, A0 = signal at time infinity, At = signal at time t
The data were fitted to equation 2) to generate Kt and kinact (inhibitor affinity driving the initial non-covalent interaction, and maximal rate of irreversible inactivation, respectively). The ratio kimcl I K, was then used as a measure of comparison.
Eqn 2 kobs = k ^a t!}— r where km3Ct = maximal rate of irreversible inactivation, [t] = concentration of compound, [L] = concentration of tracer 236, Kd - affinity of tracer236 for ITK, Ki - inhibitor affinity driving the initial non-covalent interaction. Method 2B: Assessing the reversibility of the compound : ITK complex through rapid dilution into an excess of competing iigand ("jump-dilution")
This method is necessary to complement the data generated in method 1 B, since the hyperbolic profiles generated by fitting kobs vs inhibitor concentration could also be consistent with slow-binding reversible inhibition. The protocol is essentially as for method 1 B, with the following modifications:
A 3x FAC pre-mix of 15 nM GST-ITK and 15 nM Eu-anti-GST antibody, diluted in kinase buffer A, was allowed to complex for 20 minutes at room temperature, prior to addition of 5 μΙ to 5μΙ compound in a 384-well Greiner low-volume plate. The interaction between GST-ITK-Ab pre-mix and the varying concentrations of compound was allowed to complex for 2 hours, prior to the addition of 5 μ! 900 nM (3x FAC) of kinase tracer 236. The plate was read immediately on an Envision plate reader (laser excitation, dual emission filters 665 nm and 615 nm); data were collected over a period of 6 hours at 2 minutes intervals, and the ratio of the 665 nm:615 nm emission counts was calculated. The rate of compound displacement is coincidental with the apparent increase in the FRET signal (since the rapid binding of the tracer is not the rate-limiting step). Where no apparent displacement was observed, the IC50 was calculated and compared to that predicted by equation 3), taking into account the higher (10 x Kd) concentration of the competing tracer. The ratio of the predicted IC50 vs the measured ICso was used to guide whether the data was consistent with the compound acting as an irreversible inhibitor (ie, the measured /C50was significantly more potent than predicted under these conditions).
Figure imgf000231_0001
Data on the Itk inhibition mode obtained for selection of examples: Example
22 25 28 36 39 40 number
Inhibition
Irreversible Reversible Irreversible Irreversible Irreversible Irreversible mode
Further data which can be generated using Method 1A, Method 1 B or a similar method:
Figure imgf000232_0001
Example
62 63 80 84
number
Inhibition
Irreversible Irreversible Irreversible Irreversible
mode
Ki (uM) <0.01 <0.01 <0.01 <0.01
Kinact (S 1) <0.0025 >0.0025 <0.0025 >0.0025

Claims

What is claimed is:
1. A compound of formul
Figure imgf000233_0001
(I)
wherein
R1 is hydrogen, -CH2OR4, -CH2NR6R6, -CH2-phenyl wherein the phenyl is optionally substituted by one or two substituents independently selected from Chalky!, C^alkoxy, nitrile and halo, -CH2-6-membered heterocyclyl wherein the 6-membered heterocyclyl optionally contains an oxygen atom and is optionally substituted by one or two substituents independently selected from C -6alkyl and halo, or -CH2-5-membered heteroaryl wherein the 5-membered heteroaryl contains one or two nitrogen atoms and is optionally substituted by C1-6a!kyl;
R2 is -(NH)2COCHCH2, -NR7R8, -OR9 or phenyl wherein the phenyl is optionally substituted by -NR 0R11; R3 is hydrogen, halo, Chalky! or C-i.6alkoxy;
R6 and R6 are each independently hydrogen or C^a!kyl, or
R5 and R6, together with the nitrogen atom to which they are attached, are linked to form a 5- or 6-membered heterocyclyl wherein the 6-membered heterocyclyl optionally contains an oxygen atom or a further nitrogen atom and the 5- or 6-membered heterocyclyl is optionally substituted by one or two substituents independently selected from Ci„6alkyl and halo;
R7 is hydrogen or methyl,
R8 is -COCHCH2, -(CH2)nNR12R13, C3.ecycloalkyi substituted by -OS02CH3 or -NR14R15, or 4-, 5- or 6-membered heterocyclyl wherein the 4-, 5- or 6-membered heterocyclyl contains a nitrogen atom and is substituted by -R16, or R7 and R8, together with the nitrogen atom to which they are attached, are linked to form a 6-membered heterocyc!yi optionally containing a further nitrogen atom wherein the 6- membered heterocyclyl is substituted by -R17 or -NR18R19; R9 is 5- or 6-membered heterocyclyl wherein the 5- or 6-membered heterocyclyl contains a nitrogen atom and is substituted by -R20;
R11, R 3, R 6, R16, R17, R19 and R20 are each independently -COCR25CR21R26, -COCH2halo or -COCH2OR22;
R21 is hydrogen, methyl, -CF3 or -CH2NR23R24;
R is phenyl optionally substituted by halo; R4, R10, R 2, R14, R18, R23 and R24 are each independently hydrogen or C1-6alkyl;
R2b is hydrogen, methyl or -CH2NR27R:
R is hydrogen or methyl;
R27 and R26, together with the nitrogen atom to which they are attached, are linked to form a 6-membered heterocyclyl optionally containing an oxygen atom;
X and Y are each independently -N- or -CH-; and n is 2, 3 or 4; or a salt thereof,
2. A compound according to claim 1 , or a salt thereof, wherein R1 is -CH2NR5R6.
3. A compound according to claim 1 or claim 2, or a salt thereof, wherein R2 is
NR7R8.
4. A compound according to any one of the preceding claims, or a salt thereof, wherein X is -N-.
5. A compound according to any one of the preceding claims, or a salt thereof, wherein Y is -CH-.
6. A compound selected from the group consisting of:
W-(2-{[4-[(methyioxy)methyl]-6-([1 ,33thiazoio[5,4-0]pyridin-2-yiamino)-2- pyrimidiny!]amino}ethyl)-2-propenamide;
/V-[2-({4-[(6-bromo-1 ,3-benzothiazol-2-yl)amino]-2-pyrimidinyl}amino)ethyl]-2- propenamide;
rrans-4-{[4-(4-moφholϊnylmethyl)-6-([1 ,33thiazolo[5,4·-ί)3pyπdin-2-ylarnino)-2- pyrimidinyl]amino}cyclohexyl methanesulfonate;
^-(l-acryioyl-S-piperidinyiJ-e-ICmethyloxyJmethylJ-A/^I .SJthiazolofS^-jblpyridin^-yl^^- pyrimidinediamine;
A/^il -acryloyl^-piperidiny -e-CimethyloxyJmethyll-Ar'-tl .SjthiazoloIS^-djpyridin^-yl^^- pyrimidinediamine;
ira 7S-4-{[4-[(methyloxy)methyl3-6-([1 ,3]thiazoio[5,4-/b]pyridin-2-ylamino)-2- pyrimidinyl]amino}cyclohexyl methanesulfonate;
A/-(4-{[4-[(methyloxy)methyl]-6-([1 ,3]thiazolo[5,4-6]pyridin-2-yiamino)-2- pyrimidinyl]amino}butyl)-2-propenamide;
A/-(3-{[4-[(methy!oxy)methyl]-6-([1 ,3]thiazolo[5,4-ib]pyridin-2-yiamino)-2- pyrimidinyl]amino}propyl)-2-propenamide;
/V-(fra/?s-2-{[4-[(methyloxy)methyl]-6-([1 ,3]thiazolo[5,4-0]pyridin-2-ylamino)-2- pyrimidinyijamino}cyciohexyl)-2-propenamide;
A/-(c/s-3-{[4-[(methyloxy)methyl3-6-([1 ,33thiazo!o[5,4-i)]pyndin-2-ylamino)-2- pyrimidinyl]amino}cyclohexyl)-2-propenamide;
W-(irans-3-{[4-[(methyloxy)methyi]-6-([1 ,3]thiazolo[5,4-0]pyridin-2-ylam!no)-2- pyrimidinyl]amino}cyclohexyi)-2-propenamide;
A/^ SSJ-l-acryloyl-S-pyrroltdiny -e-timethyloxyJmethyll-^ ^jthiazolofS^-bjpyndin^-yl- 2,4-pyrimidinediamine;
A/-{2-(4-acryloyl-1-piperazinyl)-6-[(methyloxy)methyl]-4-pyrimidinyl}[1 ,3]thiazolo[5,4- /b]pyridin-2-amine;
^-(^^-{^-[(methyloxyJmethylj-e-Cfl .SlthiazoloIS^-dlpyridin^-ylamino)^- pyrimidinyl]amino}cyclohexyl)-2-propenamide;
A/-(4-{[4-[(methyioxy)methyl]-6-([1 ,3]thiazolo[5,4-/}3pyridin-2-y!amino)-2- pyrimidinyi]amino}cyclohexy!)-2-propenamide;
A/-{ira/7s-4-{[4-[(methyloxy)methyl]-6-([1 ,3]thiazo!o[5,4-0]pyridin-2-ylamino)-2- pyrimidinyl]amino}cyclohexyl)-2-propenamide; A/2-[(3R)-1-acryloyl-3-pyrrolidiny!]-6-[(methy^
2,4-pyrimidinediam!ne;
A/-{2 (1-acryioyi-4-pipendinyl)oxy]-6-[(methy!oxy)methyl]-4-pyrimidinyl}[1 ,3]thiazolo[5,4 /t>]pyridin-2-amine;
(2Q-4-(dimethy]amino)- V-{2-{[4-[(methyloxy)methyl]-6-(t1 ,3]thiazolo[5 4-0]pyridin-2- y!amino)-2-pyrimidinyl]amino}ethy!)-2-butenamide;
A/2-{1 -[(2£)-4-(dimethy!amino)-2-bute^^
[1 ,3]thiazolol5,4-t»]pyridin-2-yl-2,4-pyrimidinediamine;
A/2-{1 -[(2E)-4-(dimethy!amino)-2-buten^^
[1 ,3]ihiazolo[5,4- )]pyridin-2-yl-2,4-pyrimidinediamine;
A/-(2-{[4-(4-morpholinylmethyl)-6-([1 ,33thiazo[o[5,4-/}]pyridin-2-ylamino)-2- pyrimidinyj]amino}ethyi)-2-propenamide;
W-(fra/7S-2-{[4-(4-morp olinylmethyl)-6-([1 ,3]thiazolo[5,4-ib]pyridin-2-ylamino)-2- pyrimidiny!]amino}cyciohexyl)-2-propenamide;
A/2-(1-acryloyl-3-piperidinyl)-6-(4-morpholi^
pyrimidinediamine;
A/-(frans-4-{[4-(4-morpho!inylmethyl)-6^
pyrimidinyl]amino}cyclohexyi)-2-propenamide;
W2-[(3R)-1 -acryloyi-3-pyrro!idinyi]-6-(4-^^
yl-2,4-pyrimidinediamine;
W-(c s-4-{[4-(4-morpholinylmethyl)-6-([1 ,3]thiazoio[5,4-0]pyridin-2-ylannino)-2- pyrimidiny!]amino}cyclohexyl)-2-propenamide;
/V2-(1-acryloyl-4-piperidinyl)-6-(4-morphol^
pyrimidinediamine;
A/2-[(3S)-1-acryloyl-3-pyrrolidinyl]-6-(4-m^
yl-2^pyrimidinediarnine;
A/-(c/'s-2-{[4-(4-morpho!inylmethy^
pyrimidinyl]amino}cyclohexyl)-2-propenamide;
W-(1-{6-[(6-bromo-1 ,3-benzothiazol-2-yi)amino]-4-[(2-meihyl-1 H-imidazol-1-y!)meihy!]-2- pyridinyi}-3-piperidinyl)-2-propenamide;
/V-[2-(4-acryioy!-1-piperazinyl)-6-(4-morpholinylmethyl)-4-pyrimidinyl][1 ,3]thiazolo[5,4- b]pyridin-2-arnine;
A/-[2-[(1-acryioyl-4~piperidinyl)oxy]-6-(4-morp
b]pyridin-2-amine;
A/-(c/s-3-{[4-(4-morpholinyimethyl)-6-([1 l3]thiazoioE5,4-0]pyridin-2-ylamino)-2- pyrimidinyl]amino}cyciohexyl)-2-propenamide; W-(frans-3-{[4-(4-morpholinylmethyl)-6-([1 ^
pyrimidinyl]amino}cyclohexyl)-2-propenamide;
A/z-[1-(ch!oroacetyl)-3-piperidinyl^
yl-2,4-pyrimidinediamine;
A/-[2-(2-acryloylteirahydro-1 (2H)-pyridazinyl)-6-(4-morpholinylmethyi)-4- pyrimidinyl][1 ,3]thiazoio[5,4-d]pyridin-2-amine;
/Vz-(1 -{[(4-fluorophenyl)oxy]acetyl}-3-piperidinyl)-6-(4-morphoiinylmeihyl)-A/'i- [1 ,3]thiazolo[5[4-jb]pyridin-2-yl-2,4-pyrimidinediamine;
2-chloro-/V-(2-{[4-(4-morpholinyfmethyl)-6-([1 ,3]ihiazolo[5,4- )]pyridin-2-ylamin pyrimidinyl]arriino}eihyl)acetamide;
(2£)-4,4)4-trifiuoro-A/ 2-{[4-(4-morpholinyimethyl)-6-(E1 ,3]thiazolo[5,4-6]pyridin-2- yjamino)-2-pyrimidinyl]amino}ethyl)-2-butenamide;
2 (4-fluorophenyl)oxy]-A/-(2-{[4-(4-morpholiny!methyl)-6-([1 ,3]thiazolo[5,4-0]pyridin-2- yfamino)-2-pyrimidinyl]amino}ethyl)acetamide;
Λ/-{1 -[4-(4-morpho!inylmethyi)-6-{[113]thiazo!o[5,4-jb]pyridin-2-ylamino)-2-pyridinyl]-3- piperidiny[}-2-propenamide;
W-(c s-3-{[4-(4-morpholinylmethyl)-6-([1 ,3]thiazolo[5,4-b]pyridin-2-ylamino
pyridinyl]amino}cyciohexyl)-2-propenamide;
A/-(ira/7S-3-{[4-(4-morpholinylmethyl)-6-([1 ]thtazolo[5,4-ib]pyridin-2-y!ami
pyridinyl]amino}cyclohexyl)-2-propenamide;
W-(2-{[4-(4-morpholtnylmethyl)-6-([1 l3]thiazolo[5,4-jb]pyridin-2-ylamino)-2- pyridinyl]annino}ethyi)-2-propenamide;
A/-c/s-2-{[4-(4-morpholiny!methyi)-6-([1 l3]thiazolo[5]4-b]pyridin-2-ylamino)-2 pyridiny!]amino}cyclohexyl)-2-propenamide;
A/-(irans-2-{[4-{4-morpholinylmethyl)-6-{[1 ,3]thiazolo[5,4-jb]pyridtn-2-yiamino)-^ pyridinyl3amino}cyclohexyl)-2-propenamide;
6-(4-morpholinylmethyl)-A/4-[1 ,3]thiazo!o[5^^
butenoyl]-3-piperidinyl}-2,4-pyrimidinediamine;
W-methyl-A/-(2-{[4-{4-morpho!inyim^
pyrimidinyl]amino}ethyl)-2-propenamide;
A/-{3-[4-(4-morpholinylmethyl)-6-([1 ,3]thiazolo[5^-b]pyndin-2-ylamino)-2- pyrimtdinyi]phenyl}-2-propenamide;
A/-[4-(4-morphoiinylmethyl)-6-([1 ,3]thiazolo^
propenamide;
A/'-[4-(4-morpholinyltnethyl)-6-([1 ,3]thiazolo[5,4-b]pyridin-2-ylamino)-2<1 H)- pyrimidinylidenej-2-propenohydraztde; ^-[(SSJ-l-acryloyl-S-pyrrolidiny^-A/^Ce-bromo-l ^-benzothiazol^-y!)^^- pyrimidinediamine;
W-(2-{[4-[(6-bromo-1 ,3-benzothiazol-2-yl)amino]-6-(4-morphoiinylmethyf)-2- pyrimidinyi]amino}ethyl)-2-propenamide;
/V2-[(3S)-1-acryloyl-3-pyrrolidinyl]-/V'i-(6-bromo-1 ,3-benzothiazo!-2-yl)-6-(4- morpho!inylmethyl)-2,4-pyrimidinediatnine;
W-(1 -acryloy!-4-piperidiny!)-4-(4-morpholiny!m^
pyridinediamine;
/V-[(3S)-1-acryloyl-3-pyrrolidinyi]-4-(4-m^^
y!-2,6-pyridinediamine;
A/-(1-acryloyl-3-piperidinyl)-4-(4-morpho!inylm^
pyridinediamine;
W2-(1-acryioyl-4-piperidinyl)-6-{[c/'s-2,6^
[1 ,3]thiazolo[5,4-ti]pyndin-2-yl-2l4-pyrimidinediamine;
W2-[(3S)-1 -acryloyl-3-pyrrolidinyl]-6-{[c/s-216-dimethyl-4-morpholinyl]methyl}-/V4-
[I .SJthiazoioIS^-bjpyridin^-yl^^-pyrimidinediamine;
A/-(2-{[4-{[c/s-2,6-dimethyl-4-morp^
2-pyrimidinyl]amino}ethyi)-2-propenamide;
W-<1-acry[oyl-4-piperidiny[)-4^[CTS-2,6-dim^
b]pyrtdin-2-y!-2,6-pyridinediamine;
^[(SSJ-l-acryioyl-S-pyrrolidiny^^-itc/s^^-dimethy!^-morpholinyllmethyl}-/^-
[1 ,3]thiazolo[5,4-jb]pyridin-2-y[-2,6-pyridinediamine;
W-(1 -acryloyl-3^iperidinyl)-4-{[c/s-2,6-dimethyl-4-morphoiinyl3methyl}-/V
b]pyridin-2-yl-2,6-pyridinediamine;
W-[(3S)-1 -acryloyl-3-piperidinyl]-4-{[c^
[1 ,3]thiazoto[5,4-/b3py din-2-yl-2l6-pyridinediamine;
A/-[(3R)-1 -acry!oyl-3-piperidinylj-4-{[c/s-2,6^
[1 ,3]thiazolo[5,4-i>]pyridin-2-yl-2,6-pyridinediamine;
N-{1 -[(2£)-4-(dimethylamino)-2-butenoy!]-4^
morpholinyl]methyi}-A -[1 ,33thiazoio[5,4-0]pyridin-2-yl-2,6-pyridinediamine;
A/-{(3S)-1 -[(2E)-4-(dimethyiamino)-2-butenoyl]-3-pyrrolidinyl}-4-{[c/s-2,6-dirnethyl-4- morpholinyi]methyl}-^[1 ,3]thiazolo[5^
A/-{1-[(2E)-4-{dimethy(amino)-2-butenoyl]-3-pipendinyl}-44[c/'s-2,6-dimeihyi-4- morphoiinyljmeihylJ-A/'-II .SlthiazoloIS^-^pyridin^-yl^^-pyridinediamine;
(2^-4-(dimethylamino)-A/-(2-{[4-(4-morp
ylamino)-2-pyrimidinyl]amino}ethyl)-2-butenamide; V2-{1 -[(2£)-4-(dimethyjamino)-2-butenoyl]-3-piperidinyl}-6-(4-morpholin
[1 ,3]thiazoio[5,4-0]pyridin-2-yl-2,4-pyrim!dinediamine;
W-{(3S)-1-[(2£)-4-(dimethylamino)-2-butenoyi]-3^
[1 ,3]thiazolo[5,4-jb]pyridin-2-yl-2,6-pyndinediamine;
W-{1 -[(2£)-4-(dimethylamino)-2-butenoyl]-4-piperidinyi}-4-{4-morpho!inylmeth
[1 ,3]thiazolo[5,4-b]pyridin-2-yl-2,6-pyridinediamine;
W-{1 -[(2£ -4-(dimethylamino)-2-butenoyl]-3-piperidinyl}-4-(4-morpholinylmethyl)-/V
[1 ,3]thiazolo[5,4-0]pyndin-2-y!-2,6-pyridjnediamine;
W-(1 -acry!oyl-4-piperidinyi)-4-(1 -piperidinylmet^
pyridinediamine;
A/-(1 -acryloyl-4-piperidinyl)-/V-[1.^thiazolo^-i Jpyridin^- M-tfK S)-1 ,2,2- trimethylpropyllaminoJmethy ^^-pyridinediamine;
^-(l-acryloyl^-pipendiny -W-fl .SjthiazolotS^-^pyridin^-yl^-^I R)-! ^^- trimethylpropyl]amino}methyl)-2,6-pyridinediarrtine;
W-(1 -acryloyl-4-piperidinyl)-4-[(2-methy[-1-pyrro!idinyl)methyl]-W,-[1 [3]thiazofo[5)4- i)]pyridin-2-yl-2,6-pyridinediamine;
A/-(1 ~acryioyl-4-piperidiny!)-4-[(2,2-dimethyl-4-^
6]pyridin-2-yl-2,6-pyridinediamine;
A/-[(3S)-1-acryioy!-3-pyrrolidinyl]-4-(1 -piperidinyimethyl)-A/'-[1 l3]thiazolo[5
2,6-pyridinediamine;
A/-[(3S)-1-acryloy!-3-pyrrolidinyl]-4-[(2,2-dimethyl-4-morpholinyi)methyl]-A/'- [1 ,3]thiazoio[5,4-63pyndin-2-yl-2,6-pyridinediamine;
W-[(3S)-1-acryloyl-3-pyrrolidinyl]-4-[(4-methyl-1-piperazinyl)methyl]-/\/,-[1 ,3]thiazolo[5,4- /b]pyridin-2-yl-2,6-pyridinediamine;
W<(3S)-1-acryioyl-3-pyrrolidinyl^^
trimethylpropyi]amino}methyl)-2,6-pyridinediamine;
A/-[{3S)-1-aciYloyl-3^yrrolidinyl]-AM1 ,3]thiazolo[5,4-6]pyridin-2-yl-4-({{(1 S)-1 ,2,2- trimethy!propyl]amino}methyl)-2,6-pyridinediamine;
A/-(1-acryloy!-4-piperidinyi)-4-[(dimethyiamino)m
2,6-pyridinediamine;
W-[(3S)-1 -acryloyl-3-piperidinyl]-4-{4-morp^
2,6-pyridinediamine;
W-[(3R)-1 -acry!oyl-3-pipen^inyl]-4-(4-morpto^
2,6-pyridinediamine;
4-{[c s-2,6-dimethyl-4-morphoiinyl]meihyl}-A/-[(3S)-1-(3-methyl-2-butenoyl)-3-pyrro!idinyl]-^-[I .SjthiazoloIS^-^pyridin^-yl^.e-pyridinediamine;
W-{(3S)-1 -[(2£)-2-butenoyl]-3-pyrrolidinylH-{[c s-2,6-dimethyi-4-morpholinyl]methyi}-A/'- t1 ,3]thiazolo[5,4- )]pyriciin-2-yl-2,6-pyridinediamine;
4-{[c/s-2,6-dimethyl-4-morpholinyl]m^
pyrrol!diny!}-/V 1 ,3]ihiazoloi5,4-b]pyridin-2-yl-2,6-pyridinediarnine;
4-{[c/s-2,6-dimethyl-4-morpholinyl]methyl}-A/-[(3S)-1 -(2-methylacryloy[)-3-pyrrol
[1 ,3]thiazolo[5,4-0]pyridin-2-y]-2,6-pyridinediamine;
A/-{(3S)-1-[(2£)-4-(dimethyiamtno)-2-butenoyl]-3-pyrrolidinylH [c s-2,6-dime^
morpholinyl]methyl}-A/,-[1 l33thiazoio[5,4-/b3pyridin-2-yl-2,6-pyridinediamine;
4-{[c/s-2,6-dimethyl-4-morpholinyl]m^
pyrrolidiny -A/'-fl .SjthiazolotS^-^pyridin^-yl^^-pyridinediamine;
Λ/-(1 -acryloyl-3-azetidinyl)-4-(4-morphoiinylmethyl)-/V-[1 ,3]thiazolo[5,4-jb]pyridin-2-y!-2,6- pyridtnediamine;
W-[{3S)-1 -acry!oyi-3-pyrrolidtnyl]-4-[^
2-yl-216-pyridinediamine;
A/-[(3R)-1 -acryloyl-S-pyrrolidinyn-ZV-II ,3]thiazolo[5,4-0]pyridin-2-yl-4-({[(1 S)-1 ,2,2- trimethylpropyl]amino}methyl)-2,6-pyridinediamine;
^^[(SSJ-l-acryloyi-S-pyrrolidinylJ-A^-ie-ethyl-I .S-benzothiazol^-ylJ-e-fphenylmethyl)^^- pyrimidinediamine;
/V-[(3S)-1-acry!oyl-3-pyrro!idinyl]-A/'-1 ,3-benzothiazo!-2-yl-4-(4-morphoiinylmethyl)-2,^ pyridinediamine;
A/-[(3S)-1 -acryloyl-S-pyrrolidinyll-W-^-imethyloxy)-! ,3-benzothiazol-2-yl]-4-(4- morpholinyltnethyl)-2,6-pyridinediamine;
W- SSJ-l-acryloyi-S-pyrrolidinyll-W-I .S-benzothiazoi^-yl^-^tCI SJ-l ^^- trimethylpropyllaminojmethylj^^-pyridinediamine;
/V-[(3S)-1-acryloyl-3-pyrrolidinyl]-W-t6-(methy[oxy)-1 ,3-benzothiazol-2-y]]-4-({[(1 S)-1 ,2,2- trimethyipropyl]amino}methyl)-2,6-pyridinediamine; and
W-[(3R)-1 -acry!oyl-3-pyrrolidinyl]-W-[1 ,3]thiazolo[5,4-i?]pyridin-2-yl-4-{{[(1 ,2,2- trifnethylpropy!]amino}tnethyl)-2,6-pyridinediamine; and
salts thereof.
7. A compound which is W-[(3S)-1 -acryloyl-3-pyrrolidinyl]-/V-[1 ,3]thiazoio[5,4- jb3pyndin-2-yl-4~({[(1 S)-1 ,2,2 rimethylpropyl]amino}methy!)-2,6-pyridinedianri!ne ; or a salt thereof.
8. A compound according to any one of claims 1 to 7 in the form of a pharmaceutically acceptable salt thereof.
9. W-[(3S)-1-Acryloyl-3-pyrrolidinyl]-W-[1 ,3]thiazo!o[5,4-0]pyridin-2-yl-4-({[(1 S)-1 ,2,2- trimeth lpropyl]amino}methy!)-2,6-pyridinediamine difumarate
Figure imgf000241_0001
10. A pharmaceutical composition comprising a compound as defined in any one of claims 1 to 7, or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable excipients.
1 1. A compound as defined in any one of claims 1 to 7, or a pharmaceutically acceptable salt thereof, for use in medical therapy
12. A compound as defined in any one of claims 1 to 7, or a pharmaceutically acceptable salt thereof, for use in the treatment of a disorder mediated by inappropriate Itk activity,
13. Use of a compound as defined in any one of claims 1 to 10, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for use in the treatment of a disorder mediated by inappropriate Itk activity.
14. A method of treating a disorder mediated by inappropriate Itk activity comprising administering a safe and effective amount of a compound as defined in any one of claims 1 to 7, or a pharmaceutically acceptable salt thereof, to a patient in need thereof.
15. A method according to claim 14 wherein Itk is inhibited irreversibly.
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