CA2813299A1 - Protein kinase inhibitors - Google Patents
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- CA2813299A1 CA2813299A1 CA2813299A CA2813299A CA2813299A1 CA 2813299 A1 CA2813299 A1 CA 2813299A1 CA 2813299 A CA2813299 A CA 2813299A CA 2813299 A CA2813299 A CA 2813299A CA 2813299 A1 CA2813299 A1 CA 2813299A1
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
- C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
- C07D487/04—Ortho-condensed systems
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Abstract
The present invention relates to a novel family of inhibitors of protein kinases.
In particular, the present invention relates to inhibitors of the members of the Tec and Src protein kinase families.
In particular, the present invention relates to inhibitors of the members of the Tec and Src protein kinase families.
Description
PROTEIN KINASE INHIBITORS
FIELD OF INVENTION
The present invention relates to a novel family of inhibitors of protein kinases. In particular, the present invention relates to inhibitors of the members of the Tec and Src protein kinase families.
BACKGROUND OF THE INVENTION
Protein kinases are a large group of intracellular and transmembrane signaling proteins in eukaryotic cells. These enzymes are responsible for transfer of the terminal (gamma) phosphate from ATP to specific amino acid residues of target proteins. Phosphorylation of specific tyrosine, serine or threonine amino acid residues in target proteins can modulate their activity leading to profound changes in cellular signaling and metabolism. Protein kinases can be found in the cell membrane, cytosol and organelles such as the nucleus and are responsible for mediating multiple cellular functions including metabolism, cellular growth and division, cellular signaling, modulation of immune responses, and apoptosis. The receptor tyrosine kinases are a large family of cell surface receptors with protein tyrosine kinase activity that respond to extracellular cues and activate intracellular signaling cascades (Plowman et al. (1994) DN&P, 7(6):334-339).
Aberrant activation or excessive expression of various protein kinases are implicated in the mechanism of multiple diseases and disorders characterized by benign and malignant proliferation, excess angiogenesis, as well as diseases resulting from inappropriate activation of the immune system. Thus, inhibitors of select kinases or kinase families are expected to be useful in the treatment of cancer, autoimmune diseases, and inflammatory conditions including, but not limited to: solid tumors, hematological malignancies, arthritis, graft versus host disease, lupus erythematosus, psoriasis, colitis, illeitis, multiple sclerosis, uveitis, coronary artery vasculopathy, systemic sclerosis, atherosclerosis, asthma, transplant rejection, allergy, dermatomyositis, pemphigus and the like.
Examples of kinases that can be targeted to modulate disease include receptor tyrosine kinases such as members of the platelet-derived growth factor receptor (PDGFR), vascular endothelial growth factor receptor (VEGFR) families and intracellular proteins such as members of the Syk, SRC, and Tec families of kinases.
Tec kinases are non-receptor tyrosine kinases predominantly, but not exclusively, expressed in cells of hematopoietic origin (Bradshaw W. Cell Signal. 2010,22:1175-84). The Tec family includes Tec, Bruton's tyrosine kinase (Btk), inducible T-cell kinase (Itk), resting lymphocyte kinase (RIk/Txk), and bone marrow-expressed kinase (Bmx/Etk). Btk is a Tec family kinase which is important in B-cell receptor signaling. Btk is activated by Src-family kinases and phosphorylates PLC gamma leading to effects on B-cell function and survival. Additionally, Btk is important in signal transduction in response to immune complex recognition by macrophage, mast cells and neutrophils. Btk inhibition is also important in survival of lymphoma cells (Herman, SEM. Blood 2011, 117:6287-6289) suggesting that inhibition of Btk may be useful in the treatment of lymphomas.
cSRC is the prototypical member of the SRC family of tyrosine kinases which includes Lyn, Fyn, Lck, Hck, Fgr, Blk, Syk, Yrk, and Yes. cSRC is critically involved in signaling pathways involved in cancer and is often over-expressed in human malignancies (Kim LC, Song L, Haura EB. Nat Rev Clin Oncol. 2009 6(10):587-9). The role of cSRC in cell adhesion, migration and bone remodeling strongly implicate this kinase in the development and progression of bone metastases. cSRC is also involved in signaling downstream of growth factor receptor tyrosine kinases and regulates cell cycle progression suggesting that cSRC inhibition would impact cancer cell proliferation.
Additionally, inhibition of SRC family members may be useful in treatments designed to modulate immune function. SRC family members, including Lck, regulate T-cell receptor signal transduction which leads to gene regulation events resulting in cytokine release, survival and proliferation. Thus,
FIELD OF INVENTION
The present invention relates to a novel family of inhibitors of protein kinases. In particular, the present invention relates to inhibitors of the members of the Tec and Src protein kinase families.
BACKGROUND OF THE INVENTION
Protein kinases are a large group of intracellular and transmembrane signaling proteins in eukaryotic cells. These enzymes are responsible for transfer of the terminal (gamma) phosphate from ATP to specific amino acid residues of target proteins. Phosphorylation of specific tyrosine, serine or threonine amino acid residues in target proteins can modulate their activity leading to profound changes in cellular signaling and metabolism. Protein kinases can be found in the cell membrane, cytosol and organelles such as the nucleus and are responsible for mediating multiple cellular functions including metabolism, cellular growth and division, cellular signaling, modulation of immune responses, and apoptosis. The receptor tyrosine kinases are a large family of cell surface receptors with protein tyrosine kinase activity that respond to extracellular cues and activate intracellular signaling cascades (Plowman et al. (1994) DN&P, 7(6):334-339).
Aberrant activation or excessive expression of various protein kinases are implicated in the mechanism of multiple diseases and disorders characterized by benign and malignant proliferation, excess angiogenesis, as well as diseases resulting from inappropriate activation of the immune system. Thus, inhibitors of select kinases or kinase families are expected to be useful in the treatment of cancer, autoimmune diseases, and inflammatory conditions including, but not limited to: solid tumors, hematological malignancies, arthritis, graft versus host disease, lupus erythematosus, psoriasis, colitis, illeitis, multiple sclerosis, uveitis, coronary artery vasculopathy, systemic sclerosis, atherosclerosis, asthma, transplant rejection, allergy, dermatomyositis, pemphigus and the like.
Examples of kinases that can be targeted to modulate disease include receptor tyrosine kinases such as members of the platelet-derived growth factor receptor (PDGFR), vascular endothelial growth factor receptor (VEGFR) families and intracellular proteins such as members of the Syk, SRC, and Tec families of kinases.
Tec kinases are non-receptor tyrosine kinases predominantly, but not exclusively, expressed in cells of hematopoietic origin (Bradshaw W. Cell Signal. 2010,22:1175-84). The Tec family includes Tec, Bruton's tyrosine kinase (Btk), inducible T-cell kinase (Itk), resting lymphocyte kinase (RIk/Txk), and bone marrow-expressed kinase (Bmx/Etk). Btk is a Tec family kinase which is important in B-cell receptor signaling. Btk is activated by Src-family kinases and phosphorylates PLC gamma leading to effects on B-cell function and survival. Additionally, Btk is important in signal transduction in response to immune complex recognition by macrophage, mast cells and neutrophils. Btk inhibition is also important in survival of lymphoma cells (Herman, SEM. Blood 2011, 117:6287-6289) suggesting that inhibition of Btk may be useful in the treatment of lymphomas.
cSRC is the prototypical member of the SRC family of tyrosine kinases which includes Lyn, Fyn, Lck, Hck, Fgr, Blk, Syk, Yrk, and Yes. cSRC is critically involved in signaling pathways involved in cancer and is often over-expressed in human malignancies (Kim LC, Song L, Haura EB. Nat Rev Clin Oncol. 2009 6(10):587-9). The role of cSRC in cell adhesion, migration and bone remodeling strongly implicate this kinase in the development and progression of bone metastases. cSRC is also involved in signaling downstream of growth factor receptor tyrosine kinases and regulates cell cycle progression suggesting that cSRC inhibition would impact cancer cell proliferation.
Additionally, inhibition of SRC family members may be useful in treatments designed to modulate immune function. SRC family members, including Lck, regulate T-cell receptor signal transduction which leads to gene regulation events resulting in cytokine release, survival and proliferation. Thus,
2 inhibitors of Lck have been keenly sought as immunosuppressive agents with potential application in graft rejection and T-cell mediated autoimmune disease (Martin et al. Expert Opin Ther Pat. 2010, 20:1573-93).
Inhibition of kinases using small molecule inhibitors has successfully led to several approved therapeutic agents used in the treatment of human conditions. Herein, we disclose a novel family of kinase inhibitors. Further, we demonstrate that modifications in compound substitution can influence kinase selectivity and therefore the biological function of that agent.
PCT Publication Nos. W002/080926 and W002/76986 disclose pyrazolopyrimidines as therapeutic agents. Btk is included in a long list of biologically un-related kinases. No evidence of kinase inhibition or cellular activity was disclosed in W002/080926 and exemplification centers on amide and sulfonamide derivatives with a limited subset of unsubstituted 4-phenoxyphenyl derivatives.
US Patent No. 7,514,444 discloses inhibitors of Btk. Compound 13 (PCI-32765) of this patent has been reported to show ATP competitive binding to a wide range of kinases including Btk, Lck, Lyn, cSRC, Jak, EGFR, KDR and others (Honigberg, L.A, et al, The Bruton tyrosine kinase inhibitor PCI-32765 blocks B-cell activation and is efficacious in models of autoimmune disease and B-cell malignancy,PNAS vol. 107 no. 29, 13075-13080). Specifically for Btk, the acrylannide functionality of compound 13 is reported to covalently bind the thiol moiety of Cys481, which is situated adjacent to the ATP binding pocket of Btk, thus inducing "sustained" inhibition of Btk. However, compound 13 also inhibits various kinases which also feature a Cys adjacent to the ATP binding pocket, such as Bmx, Tec, Txk, Itk, EGFR, ErbB2, ErbB4, Jak3 and Blk. Covalent binding to any of these kinases may diminish the selective nature of this approach.
CDG-0834 belongs to a structurally unrelated family of compounds which were recently reported to demonstrate significant Btk selectivity (Liu L., et al,
Inhibition of kinases using small molecule inhibitors has successfully led to several approved therapeutic agents used in the treatment of human conditions. Herein, we disclose a novel family of kinase inhibitors. Further, we demonstrate that modifications in compound substitution can influence kinase selectivity and therefore the biological function of that agent.
PCT Publication Nos. W002/080926 and W002/76986 disclose pyrazolopyrimidines as therapeutic agents. Btk is included in a long list of biologically un-related kinases. No evidence of kinase inhibition or cellular activity was disclosed in W002/080926 and exemplification centers on amide and sulfonamide derivatives with a limited subset of unsubstituted 4-phenoxyphenyl derivatives.
US Patent No. 7,514,444 discloses inhibitors of Btk. Compound 13 (PCI-32765) of this patent has been reported to show ATP competitive binding to a wide range of kinases including Btk, Lck, Lyn, cSRC, Jak, EGFR, KDR and others (Honigberg, L.A, et al, The Bruton tyrosine kinase inhibitor PCI-32765 blocks B-cell activation and is efficacious in models of autoimmune disease and B-cell malignancy,PNAS vol. 107 no. 29, 13075-13080). Specifically for Btk, the acrylannide functionality of compound 13 is reported to covalently bind the thiol moiety of Cys481, which is situated adjacent to the ATP binding pocket of Btk, thus inducing "sustained" inhibition of Btk. However, compound 13 also inhibits various kinases which also feature a Cys adjacent to the ATP binding pocket, such as Bmx, Tec, Txk, Itk, EGFR, ErbB2, ErbB4, Jak3 and Blk. Covalent binding to any of these kinases may diminish the selective nature of this approach.
CDG-0834 belongs to a structurally unrelated family of compounds which were recently reported to demonstrate significant Btk selectivity (Liu L., et al,
3 Antiarthritis effect of a novel Bruton's tyrosine kinase (BTK) inhibitor in rat collagen-induced arthritis and mechanism-based pharmacokinetic/pharmacodynamic modeling: relationships between inhibition of BTK phosphorylation and efficacy. 3 Pharmacol Exp Ther. 2011 3u1;338(1):154-63). GDC-0834 was active in several animal models of autoimmune disease. However, this compound failed in Phase 1 clinical trials as a result of human specific metabolism (Liu L, et al, Significant species difference in amide hydrolysis of GDC-0834, a novel potent and selective Bruton's tyrosine kinase inhibitor, Drug Metab Dispos. 2011 Oct;39(10):1840-9).
Inhibition of EGFR has been related to the induction of severe rash with multiple clinical compounds (Tan AR, et al, Markers in the epidermal growth factor receptor pathway and skin toxicity during erlotinib treatment. Ann Oncol. 2008 Jan;19(1):185-90). Similarly, inhibition of KDR (VEGFR2) has been clinically related to hypertension (Howard R. Mellor, et al.,Cardiotoxicity Associated with Targeting Kinase Pathways in Cancer, Toxicological Sciences 120(1), 14-32 (2011). Therefore, the development of Btk inhibitors which demonstrated greater kinase selectivity could potentially be useful in various B-cell related indications which require acute and/or chronic dosing regimens, such as cancer, inflammatory and autoimmune diseases.
The present invention relates to a family of potent and selective, non-covalent Btk inhibitors which demonstrate cellular activity, oral exposure and activity in animal models of inflammation and autoimmune disease. Kinase selectivity and cellular potency are related to specific substitution patterns on the compounds. Synthetic methods are disclosed which provide compounds on multi-gram scale.
Inhibition of EGFR has been related to the induction of severe rash with multiple clinical compounds (Tan AR, et al, Markers in the epidermal growth factor receptor pathway and skin toxicity during erlotinib treatment. Ann Oncol. 2008 Jan;19(1):185-90). Similarly, inhibition of KDR (VEGFR2) has been clinically related to hypertension (Howard R. Mellor, et al.,Cardiotoxicity Associated with Targeting Kinase Pathways in Cancer, Toxicological Sciences 120(1), 14-32 (2011). Therefore, the development of Btk inhibitors which demonstrated greater kinase selectivity could potentially be useful in various B-cell related indications which require acute and/or chronic dosing regimens, such as cancer, inflammatory and autoimmune diseases.
The present invention relates to a family of potent and selective, non-covalent Btk inhibitors which demonstrate cellular activity, oral exposure and activity in animal models of inflammation and autoimmune disease. Kinase selectivity and cellular potency are related to specific substitution patterns on the compounds. Synthetic methods are disclosed which provide compounds on multi-gram scale.
4 SUMMARY OF THE INVENTION
The present invention relates to a novel family of kinase inhibitors.
Compounds of this class have been found to have inhibitory activity against members of the Tec and Scr protein kinase families.
One aspect of the present invention is directed to a compound of Formula 1:
NH2 y_z_w I ,N
N N
iR1 Formula 1 wherein R1 is selected from the group consisting of:
1) hydrogen, 2) alkyl, 3) heteroalkyl, 4) carbocyclyl,
The present invention relates to a novel family of kinase inhibitors.
Compounds of this class have been found to have inhibitory activity against members of the Tec and Scr protein kinase families.
One aspect of the present invention is directed to a compound of Formula 1:
NH2 y_z_w I ,N
N N
iR1 Formula 1 wherein R1 is selected from the group consisting of:
1) hydrogen, 2) alkyl, 3) heteroalkyl, 4) carbocyclyl,
5) heterocyclyl,
6) -C(0)R4, wherein the alkyl, heteroalkyl, carbocyclyl and heterocyclyl may be further substituted by the groups consisting of:
1) hydroxy, 2) alkoxy, 3) alkyl, 4) -0C(0)R4, 5) -0C(0)NR5R6, 6) -C(0)R4,
1) hydroxy, 2) alkoxy, 3) alkyl, 4) -0C(0)R4, 5) -0C(0)NR5R6, 6) -C(0)R4,
7) -C(0)NR5R6,
8) -NR5R6,
9) -NR2C(0)R4,
10) -NR2S(0)nR4,
11) -NR2C(0)NR5R6;
Y is selected from:
(X2), sjsj Z is selected from:
(x1)m, Wherein -YZW is selected from:
(x2),, _________ w (X)rn.
Xl and X2 are independently selected from hydrogen, halogen or cyano;
n is an integer from 0 to 2;
m is an integer from 0 to 2;
m' is an integer from 0 to 2;
W is independently selected from:
1) alkyl, 2) aralkyl, 3) heteroaralkyl, 4) -0R3, 5) -0C(0)R4, 6) -0C(0)NR5R6, 7) -CH2O-R4, 8) -NR5R6, 9) -NR2C(0)R4, 10) -NR2S(0)nR4, 11) -NR2C(0)NR5R6;
wherein the alkyl, aralkyl and heteraralkyl may be further substituted;
R2 is selected from hydrogen or alkyl;
R3 is selected from substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted carbocyclyl, substituted or unsubstitutded heterocyclyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted aralkyl or substituted or unsubstituted heteroaralkyl;
R4 is selected from substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted carbocyclyl, substituted or unsubstitutded heterocyclyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted aralkyl or substituted or unsubstituted heteroaralkyl;
R5 and R6 are independently selected from hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, carbocyclyl, heterocyclyl, aryl, heteroaryl or R5 and R6 can be fused to form a 3 to 8 membered heterocyclyi ring system.
Preferred embodiment includes compounds of Formula 1 where W is selected from -0R3 and R3 is selected from substituted or unsubstituted aralkyl, or substituted or unsubstituted heteroaralkyl.
Preferred embodiment includes compounds of Formula 1 where W is selected from the group consisting of:
N
\\
¨0 . 1 ¨0 I/ ¨0 1 Cl,ci ¨0 , , o/ \o ¨0 . ¨0(kip // N
=____/0 41 _oy V.K
, , , , r_eil /--N ff-N
/ \NJ
/ __________________ ej / __ CS
, _0 N s /e_ D t1$
D, __I /=--- \N
/
fi , _i 0 \
D _N
1 ¨0 N
/--% ---"- c ¨0/ N -----% ---/(; 1 0 / __ el N
. k HO z--N OH
eNc // N
HO %-N/1)---/ -0 S -0 S----c I ,or I I
ií' N.
N--H .
Preferred embodiment includes compounds of Formula 1 where Fels selected from the group consisting of:
JvW
JUVV
VVVV JVNAI
...,,'' ..., .., N
II
H, CH3, acetyl, zN, D3eLCD3 ,, V-N, , (y= , =.õ..,NH
JWV
JvvV
,..../ \ .., NA0 Th\l"
N1,11 I\LIr,_, oN,I.r.0 0 0 0 H 0 , 'L) I I I I I
VVVV
JVVV
VVVV
JUVV L,.... VVVV
Ns'V'A
1\1.r. rN) c jj LI\i, n 0 0 \ ___ /, 1 III rµk 11"
I I I I I I
sss\
JVVV $4,.., srVVV VVVV
JVW .AAIV VVVV
N
NH
0 --,/ OH OH OH OH OH CI OH <>
, , , , , , , or Preferred embodiment includes compounds of Formula 1 where Y is selected from the group consisting of:
F
F
1 o . civ 1 4. 0 \
Jsrrj , ' , or .
Preferred embodiment includes compounds of Formula 1 where Z is selected from the group consisting of:
CI CN F
0\ 0 0 0 i \ l , or\
, , .
More preferred embodiment includes compounds of Formula 1 where W is selected from the group consisting of:
N
\\
* *
¨0 ¨0 , 10H, ¨N
¨0 /---C.
N ¨0 N
, or , N
--a/ _____ (s___INI
=
More preferred embodiment includes compounds of Formula 1 where RI- is selected from the group consisting of:
6,vv, õsõ,,, õõõ, iss4 VW
, JVVV
NOW JVVV
OWN!
...WV ..........s`..., VN
a _1 1 , , õ, 1 OH, H, O OH OH1 OH 0 ,or .ftrw .
More preferred embodiment includes compounds of Formula 1 where Z is selected from the group consisting of:
F
More preferred embodiment includes compounds of Formula 1 where -YZW is selected from the group consisting of:
1 lik 0 1 o Mk 0 F F or F
, .
Another aspect of the present invention provides a pharmaceutical composition comprising an effective amount of a compound of Formula 1 and a pharmaceutically acceptable carrier, diluent or excipient.
In another aspect of the present invention, there is provided a use of the compound of Formula 1 as an inhibitor of protein kinase, more particularly, as an inhibitor of Btk.
Another aspect of the present invention provides a method of modulating kinase function, the method comprising contacting a cell with a compound of the present invention in an amount sufficient to modulate the enzymatic activity of a given kinase or kinases, such as Btk, thereby modulating the kinase function.
Another aspect of the present invention provides a method of modulating the target kinase function, the method comprising a) contacting a cell with a compound of the present invention in an amount sufficient to modulate the target kinase function, thereby b) modulating the target kinase activity and signaling.
Another aspect of the present invention provides a probe, the probe comprising a compound of Formula 1 labeled with a detectable label or an affinity tag. In other words, the probe comprises a residue of a compound of Formula 1 covalently conjugated to a detectable label. Such detectable labels include, but are not limited to, a fluorescent moiety, a chemiluminescent moiety, a paramagnetic contrast agent, a metal chelate, a radioactive isotope-containing moiety, or biotin.
DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS
The present invention relates to novel kinase inhibitors. These compounds are found to have activity as inhibitors of protein kinases: including members of the tyrosine kinases Aurora, SRC (more specifically Lck) and Tec (more specifically Btk) kinase families.
Compounds of the present invention may be formulated into a pharmaceutical composition which comprises an effective amount of a compound of Formula 1 with a pharmaceutically acceptable diluent or carrier.
For example, the pharmaceutical compositions may be in a conventional pharmaceutical form suitable for oral administration (e.g., tablets, capsules, granules, powders and syrups), parenteral administration (e.g., injections (intravenous, intramuscular, or subcutaneous)), drop infusion preparations, inhalation, eye lotion, topical administration (e.g., ointment), or
Y is selected from:
(X2), sjsj Z is selected from:
(x1)m, Wherein -YZW is selected from:
(x2),, _________ w (X)rn.
Xl and X2 are independently selected from hydrogen, halogen or cyano;
n is an integer from 0 to 2;
m is an integer from 0 to 2;
m' is an integer from 0 to 2;
W is independently selected from:
1) alkyl, 2) aralkyl, 3) heteroaralkyl, 4) -0R3, 5) -0C(0)R4, 6) -0C(0)NR5R6, 7) -CH2O-R4, 8) -NR5R6, 9) -NR2C(0)R4, 10) -NR2S(0)nR4, 11) -NR2C(0)NR5R6;
wherein the alkyl, aralkyl and heteraralkyl may be further substituted;
R2 is selected from hydrogen or alkyl;
R3 is selected from substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted carbocyclyl, substituted or unsubstitutded heterocyclyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted aralkyl or substituted or unsubstituted heteroaralkyl;
R4 is selected from substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted carbocyclyl, substituted or unsubstitutded heterocyclyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted aralkyl or substituted or unsubstituted heteroaralkyl;
R5 and R6 are independently selected from hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, carbocyclyl, heterocyclyl, aryl, heteroaryl or R5 and R6 can be fused to form a 3 to 8 membered heterocyclyi ring system.
Preferred embodiment includes compounds of Formula 1 where W is selected from -0R3 and R3 is selected from substituted or unsubstituted aralkyl, or substituted or unsubstituted heteroaralkyl.
Preferred embodiment includes compounds of Formula 1 where W is selected from the group consisting of:
N
\\
¨0 . 1 ¨0 I/ ¨0 1 Cl,ci ¨0 , , o/ \o ¨0 . ¨0(kip // N
=____/0 41 _oy V.K
, , , , r_eil /--N ff-N
/ \NJ
/ __________________ ej / __ CS
, _0 N s /e_ D t1$
D, __I /=--- \N
/
fi , _i 0 \
D _N
1 ¨0 N
/--% ---"- c ¨0/ N -----% ---/(; 1 0 / __ el N
. k HO z--N OH
eNc // N
HO %-N/1)---/ -0 S -0 S----c I ,or I I
ií' N.
N--H .
Preferred embodiment includes compounds of Formula 1 where Fels selected from the group consisting of:
JvW
JUVV
VVVV JVNAI
...,,'' ..., .., N
II
H, CH3, acetyl, zN, D3eLCD3 ,, V-N, , (y= , =.õ..,NH
JWV
JvvV
,..../ \ .., NA0 Th\l"
N1,11 I\LIr,_, oN,I.r.0 0 0 0 H 0 , 'L) I I I I I
VVVV
JVVV
VVVV
JUVV L,.... VVVV
Ns'V'A
1\1.r. rN) c jj LI\i, n 0 0 \ ___ /, 1 III rµk 11"
I I I I I I
sss\
JVVV $4,.., srVVV VVVV
JVW .AAIV VVVV
N
NH
0 --,/ OH OH OH OH OH CI OH <>
, , , , , , , or Preferred embodiment includes compounds of Formula 1 where Y is selected from the group consisting of:
F
F
1 o . civ 1 4. 0 \
Jsrrj , ' , or .
Preferred embodiment includes compounds of Formula 1 where Z is selected from the group consisting of:
CI CN F
0\ 0 0 0 i \ l , or\
, , .
More preferred embodiment includes compounds of Formula 1 where W is selected from the group consisting of:
N
\\
* *
¨0 ¨0 , 10H, ¨N
¨0 /---C.
N ¨0 N
, or , N
--a/ _____ (s___INI
=
More preferred embodiment includes compounds of Formula 1 where RI- is selected from the group consisting of:
6,vv, õsõ,,, õõõ, iss4 VW
, JVVV
NOW JVVV
OWN!
...WV ..........s`..., VN
a _1 1 , , õ, 1 OH, H, O OH OH1 OH 0 ,or .ftrw .
More preferred embodiment includes compounds of Formula 1 where Z is selected from the group consisting of:
F
More preferred embodiment includes compounds of Formula 1 where -YZW is selected from the group consisting of:
1 lik 0 1 o Mk 0 F F or F
, .
Another aspect of the present invention provides a pharmaceutical composition comprising an effective amount of a compound of Formula 1 and a pharmaceutically acceptable carrier, diluent or excipient.
In another aspect of the present invention, there is provided a use of the compound of Formula 1 as an inhibitor of protein kinase, more particularly, as an inhibitor of Btk.
Another aspect of the present invention provides a method of modulating kinase function, the method comprising contacting a cell with a compound of the present invention in an amount sufficient to modulate the enzymatic activity of a given kinase or kinases, such as Btk, thereby modulating the kinase function.
Another aspect of the present invention provides a method of modulating the target kinase function, the method comprising a) contacting a cell with a compound of the present invention in an amount sufficient to modulate the target kinase function, thereby b) modulating the target kinase activity and signaling.
Another aspect of the present invention provides a probe, the probe comprising a compound of Formula 1 labeled with a detectable label or an affinity tag. In other words, the probe comprises a residue of a compound of Formula 1 covalently conjugated to a detectable label. Such detectable labels include, but are not limited to, a fluorescent moiety, a chemiluminescent moiety, a paramagnetic contrast agent, a metal chelate, a radioactive isotope-containing moiety, or biotin.
DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS
The present invention relates to novel kinase inhibitors. These compounds are found to have activity as inhibitors of protein kinases: including members of the tyrosine kinases Aurora, SRC (more specifically Lck) and Tec (more specifically Btk) kinase families.
Compounds of the present invention may be formulated into a pharmaceutical composition which comprises an effective amount of a compound of Formula 1 with a pharmaceutically acceptable diluent or carrier.
For example, the pharmaceutical compositions may be in a conventional pharmaceutical form suitable for oral administration (e.g., tablets, capsules, granules, powders and syrups), parenteral administration (e.g., injections (intravenous, intramuscular, or subcutaneous)), drop infusion preparations, inhalation, eye lotion, topical administration (e.g., ointment), or
12 suppositories. Regardless of the route of administration selected the compounds may be formulated into pharmaceutically acceptable dosage forms by conventional methods known to those skilled in the art.
The phrase "pharmaceutically acceptable" is employed herein to refer to those ligands, materials, compositions, and/or 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, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.
The phrase "pharmaceutically acceptable carrier" as used herein means a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material.
Each carrier must be acceptable in the sense of being compatible with the other ingredients of the formulation, including the active ingredient, and not injurious or harmful to the patient. Some examples of materials which can serve as pharmaceutically acceptable carriers include: (1) sugars, such as lactose, glucose, and sucrose; (2) starches, such as corn starch, potato starch, and substituted or unsubstituted 8-cyclodextrin; (3) cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc;
(8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerin, sorbitol, mannitol, and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffering agents, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethyl alcohol; (20) phosphate buffer solutions; and (21) other non-toxic compatible substances employed in pharmaceutical formulations.
The term "pharmaceutically acceptable salt" refers to the relatively non-toxic, inorganic and organic acid addition salts of the compound(s). These salts
The phrase "pharmaceutically acceptable" is employed herein to refer to those ligands, materials, compositions, and/or 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, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.
The phrase "pharmaceutically acceptable carrier" as used herein means a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material.
Each carrier must be acceptable in the sense of being compatible with the other ingredients of the formulation, including the active ingredient, and not injurious or harmful to the patient. Some examples of materials which can serve as pharmaceutically acceptable carriers include: (1) sugars, such as lactose, glucose, and sucrose; (2) starches, such as corn starch, potato starch, and substituted or unsubstituted 8-cyclodextrin; (3) cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc;
(8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerin, sorbitol, mannitol, and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffering agents, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethyl alcohol; (20) phosphate buffer solutions; and (21) other non-toxic compatible substances employed in pharmaceutical formulations.
The term "pharmaceutically acceptable salt" refers to the relatively non-toxic, inorganic and organic acid addition salts of the compound(s). These salts
13 can be prepared in situ during the final isolation and purification of the compound(s), or by separately reacting a purified compound(s) in its free base form with a suitable organic or inorganic acid, and isolating the salt thus formed. Representative salts include the hydrobromide, hydrochloride, sulfate, bisulfate, phosphate, nitrate, acetate, valerate, oleate, palmitate, stearate, laurate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, naphthylate, mesylate, glucoheptonate, lactobionate, laurylsulphonate salts, and amino acid salts, and the like (See, for example, Berge et al. (1977) "Pharmaceutical Salts", J. Pharm. Sci. 66:
1-19).
In other cases, the compounds of the present invention may contain one or more acidic functional groups and, thus, are capable of forming pharmaceutically acceptable salts with pharmaceutically acceptable bases.
The term "pharmaceutically acceptable salts" in these instances refers to the relatively non-toxic inorganic and organic base addition salts of a compound(s). These salts can likewise be prepared in situ during the final isolation and purification of the compound(s), or by separately reacting the purified compound(s) in its free acid form with a suitable base, such as the hydroxide, carbonate, or bicarbonate of a pharmaceutically acceptable metal cation, with ammonia, or with a pharmaceutically acceptable organic primary, secondary, or tertiary amine. Representative alkali or alkaline earth salts include the lithium, sodium, potassium, calcium, magnesium, and aluminum salts, and the like. Representative organic amines useful for the formation of base addition salts include ethylamine, diethylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine, and the like (see, for example, Berge et al., supra).
As used herein, the term "affinity tag" means a ligand or group, linked either to a compound of the present invention or to a protein kinase domain, that allows the conjugate to be extracted from a solution.
The term "alkyl" refers to substituted or unsubstituted saturated hydrocarbon groups, including straight-chain alkyl and branched-chain alkyl groups,
1-19).
In other cases, the compounds of the present invention may contain one or more acidic functional groups and, thus, are capable of forming pharmaceutically acceptable salts with pharmaceutically acceptable bases.
The term "pharmaceutically acceptable salts" in these instances refers to the relatively non-toxic inorganic and organic base addition salts of a compound(s). These salts can likewise be prepared in situ during the final isolation and purification of the compound(s), or by separately reacting the purified compound(s) in its free acid form with a suitable base, such as the hydroxide, carbonate, or bicarbonate of a pharmaceutically acceptable metal cation, with ammonia, or with a pharmaceutically acceptable organic primary, secondary, or tertiary amine. Representative alkali or alkaline earth salts include the lithium, sodium, potassium, calcium, magnesium, and aluminum salts, and the like. Representative organic amines useful for the formation of base addition salts include ethylamine, diethylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine, and the like (see, for example, Berge et al., supra).
As used herein, the term "affinity tag" means a ligand or group, linked either to a compound of the present invention or to a protein kinase domain, that allows the conjugate to be extracted from a solution.
The term "alkyl" refers to substituted or unsubstituted saturated hydrocarbon groups, including straight-chain alkyl and branched-chain alkyl groups,
14 including haloalkyl groups such as trifluoromethyl and 2,2,2-trifluoroethyl, etc. Representative alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, (cyclohexyl)methyl, cyclopropylmethyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, and the like. The terms "alkenyl" and "alkynyl" refer to substituted or unsubstituted unsaturated aliphatic groups analogous in length and possible substitution to the alkyls described above, but that contain at least one double or triple bond respectively.
Representative alkenyl groups include vinyl, propen-2-yl, crotyl, isopenten-2-yl, 1,3-butadien-2-y1), 2,4-pentadienyl, and 1,4-pentadien-3-yl.
Representative alkynyl groups include ethynyl, 1- and 3-propynyl, and 3-butynyl. In certain preferred embodiments, alkyl substituents are lower alkyl groups, e.g., having from 1 to 6 carbon atoms. Similarly, alkenyl and alkynyl preferably refer to lower alkenyl and alkynyl groups, e.g., having from 2 to 6 carbon atoms. As used herein, "alkylene" refers to an alkyl group with two open valencies (rather than a single valency), such as -(Ch12)1-10- and substituted variants thereof.
The term "alkoxy" refers to an alkyl group having an oxygen attached thereto. Representative alkoxy groups include methoxy, ethoxy, propoxy, tert-butoxy and the like. An "ether" is two hydrocarbons covalently linked by an oxygen. Accordingly, the substituent of an alkyl that renders that alkyl an ether is or resembles an alkoxy.
The term "alkoxyalkyl" refers to an alkyl group substituted with an alkoxy group, thereby forming an ether.
The terms "amide" and "amido" are art-recognized as an amino-substituted carbonyl and includes a moiety that can be represented by the general formula:
A ,R10 N
i wherein R9, le) are as defined above. Preferred embodiments of the amide will not include imides, which may be unstable.
The terms "amine" and "amino" are art-recognized and refer to both unsubstituted and substituted amines and salts thereof, e.g., a moiety that can be represented by the general formulae:
1 .,, ¨14 or ¨N4.¨R ''' i \R10 Rio' wherein R9, 111 and R1 ' each independently represent a hydrogen, an alkyl, an alkenyl, -(CH2)m-R8, or R9 and Rw taken together with the N atom to which they are attached complete a heterocycle having from 4 to 8 atoms in the ring structure; R8 represents an aryl, a cycloalkyl, a cycloalkenyl, a heterocyclyl or a polycyclyl; and m is zero or an integer from 1 to 8. In preferred embodiments, only one of R9 or Ftw can be a carbonyl, e.g., R9, R10, and the nitrogen together do not form an imide. In even more preferred embodiments, R9 and le) (and optionally R1w) each independently represent a hydrogen, an alkyl, an alkenyl, or -(CH2)m-R8. In certain embodiments, the amino group is basic, meaning the protonated form has a pKa > 7.00.
The term "aralkyl", as used herein, refers to an alkyl group substituted with an aryl group, for example -(CH2)n-Ar.
The term "heteroaralkyl", as used herein, refers to an alkyl group substituted with a heteroaryl group, for example -(CH2)n-Het.
The term "aryl" as used herein includes 5-, 6-, and 7-membered substituted or unsubstituted single-ring aromatic groups in which each atom of the ring is carbon. The term "aryl" also includes polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjoining rings wherein at least one of the rings is aromatic, e.g., the other cyclic rings can be cycloalkyls, cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and/or heterocyclyls. Aryl groups include benzene, naphthalene, phenanthrene, phenol, aniline, anthracene, and phenanthrene.
The terms "carbocycle" and "carbocyclyl", as used herein, refer to a non-aromatic substituted or unsubstituted ring in which each atom of the ring is carbon. The terms "carbocycle" and "carbocycly1" also include polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjoining rings wherein at least one of the rings is carbocyclic, e.g., the other cyclic rings can be cycloalkyls, cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and/or heterocyclyls. Representative carbocyclic groups include cyclopentyl, cyclohexyl, 1-cyclohexenyl, and 3-cyclohexen-1-yl, cycloheptyl.
The term "carbonyl" is art-recognized and includes such moieties as can be represented by the general formula:
AR"
X' wherein X is a bond or represents an oxygen or a sulfur, and Rn represents a hydrogen, an alkyl, an alkenyl, -(CH2)m-R8 or a pharmaceutically acceptable salt. Where X is an oxygen and R11 is not hydrogen, the formula represents an "ester". Where X is an oxygen, and Rfi is a hydrogen, the formula represents a "carboxylic acid".
The terms "heteroaryl" includes substituted or unsubstituted aromatic 5- to 7-membered ring structures, more preferably 5- to 6-membered rings, whose ring structures include one to four heteroatoms. The term "heteroaryl" also includes polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjoining rings wherein at least one of the rings is heteroaromatic, e.g., the other cyclic rings can be cycloalkyls, cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and/or heterocyclyls. Heteroaryl groups include, for example, pyrrole, furan, thiophene, imidazole, isoxazole, oxazole, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazine and pyrimidine, and the like.
The term "heteroatom" as used herein means an atom of any element other than carbon or hydrogen. Preferred heteroatoms are nitrogen, oxygen, and sulfur.
The terms "heterocycly1" or "heterocyclic group" refer to substituted or unsubstituted non-aromatic 3- to 10-membered ring structures, more preferably 3- to 7-membered rings, whose ring structures include one to four heteroatoms. The term terms "heterocyclyl" or "heterocyclic group" also include polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjoining rings wherein at least one of the rings is heterocyclic, e.g., the other cyclic rings can be cycloalkyls, cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and/or heterocyclyls.
Heterocyclyl groups include, for example, tetrahydrofuran, tetrahydropyran, piperidine, piperazine, pyrrolidine, morpholine, lactones, and lactams.
The term "hydrocarbon", as used herein, refers to a group that is bonded through a carbon atom that does not have a =0 or =S substituent, and typically has at least one carbon-hydrogen bond and a primarily carbon backbone, but may optionally include heteroatoms. Thus, groups like methyl, ethoxyethyl, 2-pyridyl, and trifluoromethyl are considered to be hydrocarbyl for the purposes of this application, but substituents such as acetyl (which has a =0 substituent on the linking carbon) and ethoxy (which is linked through oxygen, not carbon) are not. Hydrocarbyl groups include, but are not limited to aryl, heteroaryl, carbocycle, heterocycle, alkyl, alkenyl, alkynyl, and combinations thereof.
The terms "polycycly1" or "polycyclic" refer to two or more rings (e.g., cycloalkyls, cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and/or heterocyclyls) in which two or more carbons are common to two adjoining rings, e.g., the rings are "fused rings". Each of the rings of the polycycle can be substituted or unsubstituted.
As used herein, the term "probe" means a compound of the invention which is labeled with either a detectable label or an affinity tag, and which is capable of binding, either covalently or non-covalently, to a protein kinase domain. When, for example, the probe is non-covalently bound, it may be displaced by a test compound. When, for example, the probe is bound covalently, it may be used to form cross-linked adducts, which may be quantified and inhibited by a test compound.
The term "substituted" refers to moieties having substituents replacing a hydrogen on one or more carbons of the backbone. It will be understood that "substitution" or "substituted with" includes the implicit proviso that such substitution is in accordance with permitted valence of the substituted atom and the substituent, and that the substitution results in a stable compound, e.g., which does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, etc. As used herein, the term "substituted" is contemplated to include all permissible substituents of organic compounds. In a broad aspect, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of organic compounds. The permissible substituents can be one or more and the same or different for appropriate organic compounds. For purposes of this invention, the heteroatoms such as nitrogen may have hydrogen substituents and/or any permissible substituents of organic compounds described herein which satisfy the valences of the heteroatoms. Substituents can include, for example, a halogen, a hydroxyl, a carbonyl (such as a carboxyl, an alkoxycarbonyl, a formyl, or an acyl), a thiocarbonyl (such as a thioester, a thioacetate, or a thioformate), an alkoxyl, a phosphoryl, a phosphate, a phosphonate, a phosphinate, an amino, an amido, an amidine, an imine, a cyano, a nitro, an azido, a sulfhydryl, an alkylthio, a sulfate, a sulfonate, a sulfamoyl, a sulfonamido, a sulfonyl, a heterocyclyl, an aralkyl, or an aromatic or heteroaromatic moiety. It will be understood by those skilled in the art that the moieties substituted on the hydrocarbon chain can themselves be substituted, if appropriate.
Compounds of the invention also include all isotopes of atoms present in the intermediates and/or final compounds. Isotopes include those atoms having the same atomic number but different mass numbers. For example, isotopes of hydrogen include deuterium and tritium.
General Synthetic Methods General Synthetic Method A:
Ulmann condensation of phenol 1-i with ester 1-ii provided intermediate 1-iii.
Saponification of intermediate 1-iii yielded intermediate 1-iv. Conversion of intermediate 1-iv to its acid chloride, using for example oxalyl chloride and DMF, provided intermediate 1-v. Condensation of intermediate 1-v with malononitrile yielded intermediate 1-vi. Methylation of intermediate 1-vi with TMS-diazomethane provided intermediate 1-vii. Condensation of 1-vii with hydrazine yielded intermediate 1-viii. Condensation of intermediate 1-viii with formamidine yielded intermediate 1-ix. Intermediate 1-ix was treated with alcohol R101-I, under Mitsunobu conditions, to provide the desired compounds or intermediates of general formula 1-x.
(X16. (X16 (X16.
\ ,õ...,,,vv Base, ligand, 1\'-/-w TW
1 catalyst /I- I
..f- NaOH
______________________________________________ ), Y (x2)m OH 0 0 OH
7C) EtO2C (X26 HO2C X2), 1-iii 1-iv 1-ii; X=I, Br (X1),õ. (X1)m.
(x1),,=
t71\1 o W
oxalyl chloride --' malononitrile .vi'm TMS-diazomethane ,j7W
1-iv i ¨(X2)m ' I ¨(X2)rn 1 \
CIOC)(X2)m HOCN Me0 CN
CN CN
1-v 1-vi 1-vii _<------- )m, 0 \ ?
hydrazine / \ W
1-vii _______________________ .
NC -----0(2)m / \,NI
H
1-viii (X1) . (Xl)m 0____.
a m z' c-- \ /
Ph3P, DIAD
W W
.. o. f rmamidine / \
1-viii _______ is. _________________________ 1 NH2 ' (X2)m NH2 ----(X2)m RiOH
\ N N
N ' N I :
vi 1-ix 1-x Scheme 1 General Synthetic Method 13:
Benzoyl chlorides of formula 2-1 were condensed with malononitrile to provide intermediate 2-11. Methylation of intermediate 2-11 with TMS-diazomethane provided intermediate 2-iii. Condensation of intermediate 2-iii with hydrazine provided intermediate 2-iv. Further condensation of intermediate 2-iv with formamidine provided intermediate 2-v. Intermediate 2-v was treated with alcohol R1OH, under Mitsunobu conditions, to provide intermediate 2-vi. Ullmann condensation of intermediate 2-vi with phenolic intermediates 2-vii provided the desired compounds or intermediates of general formula 2-viii.
X
X
X
X
TMS-diazomethane /1-=
.L1 )(2,r, malononitrile rjTh.._(x2) 1 ___(x26 hydrazine ( ) CN o CN NC--.)N
0 Cl HO-1-' CN NH
.'' 2-1; X=1, Br 2-ii; X=1, Br 2-111; X=1, Br 2-iv; X=1, Br X
X / \
formamidine / \ Ph3P, D1AD, 2-iv NH2 -------(X-, )171 IsV , \
1:210H
N \ N N, ' R1 N N
2-v; X=1, Br 2-vi; X=1, Br 0 \ /
Base, ligand, catalyst 2-vi _____________________ p.
NH2 --(X2)m HOW
1 N \
1 ,N
(X1),õ, N N, 2-vii 2-viii Scheme 2a Alternatively, intermediate 2-iv was treated with an alcohol of formula R101-1, under Mitsunobu conditions to provide intermediate 2-ix. Condensation of intermediate 2-ix with formamidine provided intermediate 2-vi.
X
X X
NC Ph3P, DIAD, NC (X26 formamidine NH2 ________________________________________________ k \ RiOH
N N
NH I \ N
H2NN,/
2-iv; X=1, Br 2-ix; X=1, Br 2-vi; X=1, Br Scheme 2b In a similar manner, condensation of intermediate 2-iii with a hydrazine of formula R1NIHNH2 provided intermediate 2-ix. Ullmann condensation of intermediate 2-ix with phenolic intermediates 2-vii provided intermediates 2-x. Condensation of intermediate 2-x with formamidine provided the desired compounds or intermediates of general formula 2-viii.
Xa(xi)m.
, \ 0-7 _(X2),, Base, ligand, \ W
-H1-c2N6 RiNHNH2 NC Ho w . \
NC(X2)m \
CN Ri /'" H HN14 --(X1),,, Ri 2-iii; X=1, Br 2-ix; X=1, Br 2-vii 2-x -(X1)m' 0_ \ /
W
/ \ 2 formamidine 2-x ________________________ .
N \ N
I ' le---N
Ft1 2-viii Scheme 2c Alternatively, trimethyl orthoformate and ammonia can be used in place of formamidine, for example, in the conversion of intermediate 2-x to compounds of formula 2-vii.
Exemplification The following synthetic methods are intended to be representative of the chemistry used to prepare compounds of Formula 1 and are not intended to be limiting.
Synthesis of Compound 1:
0 OBn OH K2CO3 0 OBn CuCI, Cs2CO3 ___________________ r _____________________ ).
Benzyl bromide 0 0 OH OH 40 o'' Ir EtO2C
3-a Br 3-b 0 OBn 0 OBn NaOH oxalyl chloride 3-b __________ r id 0 _________ N ja 0 Mr ir 3-c 3-d o IS o 010 3-d DIPEA 0 TMS-diazomethane 0 malononitrile 0 40 '-.
HO Me0 CN --. CN
CN CN
3-e 3-f 0 * O, hydrazine 41 NC 0 formamidine 3-f r ___________________________ r i \isl . N '' , \ it H2N N, L i ,N
H N N
H
3-g Compound 1 Scheme 3 Step 1: Intermediate 3-a Benzyl bromide (27.0 ml, 227 mmol) was added drop wise to a stirred suspension of resorcinol (25.0 g, 227 mmol) and potassium carbonate (31.4 g, 227 mmol) in acetone (150 ml) and the reaction was heated under reflux overnight. Volatiles were removed under reduced pressure. Water and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure.
Purification by silica gel chromatography provided intermediate 3-a as a beige oil.
Step 2: Intermediate 3-b To a solution of compound 3-a (15.0 g, 74.9 mmol) in 1,4-dioxane (200 ml) were sequentially added ethyl 4-bromobenzoate (20.59 g, 90 mmol), N,N-dimethylglycine (4.25 g, 41.2 mmol), copper(I) chloride(3.71 g, 37.5 mmol) and cesium carbonate (61.0 g, 187 mmol). The reaction mixture was stirred at reflux overnight and then cooled to room temperature. Water and ethyl acetate were added, the organic layer was separated, washed with saturated aqueous NaHCO3, brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 3-b as a colorless oil.
Step 3: Intermediate 3-c To a solution of intermediate 3-b (17.5 g, 50.2 mmol) in THF (200 ml) and Me0H (100 ml) was added 2N sodium hydroxide (100 ml, 200 mmol) and the reaction was stirred at room temperature overnight. Volatiles were removed under reduced pressure. 10% aqueous HCI and ethyl acetate were added to the residue, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure to provide intermediate 3-c as beige solid.
Step 4: Intermediate 3-d To a suspension of intermediate 3-c (16.1 g, 50.3 mmol) in dichloromethane (100 ml) were added DMF (0.1 ml, 1.29 mmol) and oxalyl chloride (4.4 ml, 50.3 mmol). The solution was stirred at room temperature for 2 hours.
Volatiles were removed under reduced pressure to provide intermediate 3-d as beige solid.
Step 5: Intermediate 3-e To a solution of intermediate 3-d (16.5 g, 48.9 mmol) in toluene (50 ml) and THF (7 ml), cooled to -10 C, were added malononitrile (3.19 ml, 50.2 mmol) and DIPEA (17.5 ml, 100 mmol) in toluene (50 mL), drop wise, over a period of 30 minutes. After the addition was completed, the reaction was stirred for 1 hour at 0 C and room temperature overnight. Volatiles were removed under reduced pressure. 1M aqueous HCI and ethyl acetate were added, the organic layer was separated, washed with 1M HCI and brine, dried over MgSO4, filtered and concentrated under reduced pressure to provide intermediate 3-e as beige solid.
Step 6: Intermediate 3-f To a solution of intermediate 3-e (18.1 g, 49.1 mmol) in acetonitrile (177 ml) and methanol (19.0 ml), cooled to 0 C, were added DIPEA (10.3 ml, 59.0 mmol) and a 2M solution of (diazomethyl)trimethylsilane in hexanes (27.0 ml, 54.0 mmol). After the addition was completed, the reaction was stirred at room temperature overnight. Acetic acid (0.56 ml, 9.83 mmol) was added, the reaction was then stirred for 30 minutes and volatiles were removed under reduced pressure. A saturated aqueous solution of NaHCO3 and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure.
Purification by silica gel chromatography provided intermediate 3-f as yellow solid.
Step 7: Intermediate 3-g To a suspension of intermediate 3-f (8.05 g, 21.1 mmol) in ethanol (10.5 ml) was added a solution of hydrazine monohydrate (2.76 ml, 56.8 mmol). The reaction was stirred at 100 C for 1 hour and then cooled to room temperature.= Water was added; a precipitate formed and was collected by filtration, washed with diethyl ether and dried in vacuo to provide intermediate 3-g as an off-white solid.
Step 8: Compound 1 Intermediate 3-g (8.0 g, 20.92 mmol) was added to a solution of formamidine (58.4 ml, 1464 mmol) and the reaction was stirred at 180 C
for 2 hours and then cooled to room temperature. Water and ethyl acetate were added; the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure to provide compound 1 as beige solid. MS (m/z) M+H=410.2 Synthesis of Compound 2:
O . o 40 .
o o NH2 41k 40 Ph3P, DIAD NH2 N' \N N
OH N
i N' [.., 1 )1 N
H
uCompound 1 Compound 2 Scheme 4 To a solution of cyclopentanol (316 mg, 3.66 mmol) in THF was added triphenylphosphine (961 mg, 3.66 mmol) and DIAD (712 pl, 3.66 mmol). The yellow solution was stirred for 5 minutes, compound 1 (1.0 g, 2.44 mmol) was added and the reaction was then stirred at room temperature overnight.
Volatiles were removed under reduced pressure. Purification by silica gel chromatography provided compound 2 as an off-white solid. MS (m/z) M+H=478.2 Synthesis of Compound 3:
, o 0 NH2 * Ph3P, DIAD
N
N 41#
N
' N
N N' Ny(31 N N
Compound 1 Compound 3 Scheme 5 To a solution of (S)-tert-butyl 3-hydroxypiperidine-1-carboxylate (5.65 g, 28.1 mmol) in THF was added triphenylphosphine (7.37 g, 28.1 mmol) and DIAD (5.46 ml, 28.1 mmol). The yellow solution was stirred for 5 minutes, compound 1 (10.0 g, 24.42 mmol) was added and the reaction was then stirred at room temperature overnight. Volatiles were removed under reduced pressure. Purification by silica gel chromatography provided compound 3 as a white foam. MS (m/z) M+H= 593.1 Synthesis of Compound 4:
o NH2 41, HCI
NH2 *
\ N
I N I ,N
N N N
LNH
Compound 4 Compound 3 Scheme 6 To a solution of compound 3 (1.88 g, 3.17 mmol) in dichloromethane was added 4N HCI in 1,4-dioxane (19.82 ml, 79.0 mmol) and the reaction was stirred at room temperature for 2 hours. Volatiles were removed under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 4.2HCI as a white solid. MS (m/z) M+H= 493.1 Synthesis of compound 5 0 = o, TEA
4Ik 0 NH2 N \
'N I N
òH
Compound 4 Compound 5 Scheme 7 To a solution of compound 4.2HCI (100 mg, 0.17 mmol) in dichloromethane (2 ml) cooled to 0 C were sequentially added TEA (99 pl, 0.70 mmol) and acryloyl chloride (17.6 mg, 0.19 mmol). The reaction was stirred at 0 C for 1 hour. A saturated aqueous solution of ammonium chloride was added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided compound 5 as a white solid. MS (m/z) M+H=
547.1 Synthesis of compound 6 ifh o, TEA
0 =
41kN \ 11 CI N \ N
I , N'N N
L\NH LN
Compound 4 Compound 6 Scheme 8 To a solution of compound 4.2HCI (1.8 g, 3.18 mmol) in dichloromethane (32 ml) cooled to 0 C were sequentially added TEA (1.77 ml, 12.73 mmol) and acetyl chloride (249 pl, 3.50 mmol). The reaction was stirred at 0 C for 1 hour and room temperature overnight. A saturated aqueous solution of ammonium chloride was added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with 1%
aqueous HCl/methanol gradient provided compound 6.11C1 as beige solid. MS
(m/z) M+H= 535.1 Synthesis of intermediate 9-d Br Br Br 0 DIPEA , malononitrile (110 TMS-diazomethane SI
________________________________________________ A
CN =-. \ CN
HO
0 CI , CN
CN
9-a 9-b Br Br hydrazine 0 formamidine 9-b __________ A ' NH2 fa NC \'N N \ N
NH N N
H
9-c 9-d Scheme 9 Step 1: Intermediate 9-a To a solution of 4-bromobenzoyl chloride (25.0 g, 114 mmol) in toluene (200 ml) and THF (30 ml), cooled to -10 C, were sequentially added malononitrile (7.60 ml, 120.0 mmol) and DIPEA (39.8 ml, 228 mmol) in toluene (50 mL) drop wise over a period of 1 hour. After the addition was completed, the reaction was stirred for 1 hour at 0 C and room temperature overnight.
Volatiles were removed under reduced pressure. 1N HCI and ethyl acetate were added to the residue, the organic layer was separated, washed twice with 1N HCI and brine, dried over MgSO4, filtered and concentrated under reduced pressure to provide intermediate 9-a as yellow solid.
Step 2: Intermediate 9-b To a solution of intermediate 9-a (26.4 g, 106 mmol) in acetonitrile (300 ml) and methanol (35.0 ml), cooled to 0 C, was added DIPEA (22.2 ml, 127 mmol) and a 2M solution of diazomethyl)trimethylsilane in hexanes (58.3 ml, 117 mmol). After the addition was completed, the reaction was stirred at room temperature overnight. Acetic acid (1.21 ml, 21.2 mmol) was added, the reaction was stirred for 30 minutes and volatiles were removed under reduced pressure. A saturated aqueous solution of NaHCO3 and ethyl acetate were added, the organic layer was separated, washed with brine, dried over M9SO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 9-b as a yellow solid.
Step 3: Intermediate 9-c To a suspension of intermediate 9-b (4.49 g, 17.07 mmol) in ethanol (8.5 ml) was added a solution of hydrazine monohydrate (2.23 ml, 46.1 mmol) and the reaction was stirred at 100 C for 1 hour and then cooled to room temperature. Volatiles were removed under reduced pressure to provide intermediate 9-c as a yellow solid.
Step 4: Intermediate 9-d Intermediate 9-c (4.49 g, 17.07 mmol) was added to a solution of formamidine (40.8 ml, 1024 mmol) and the reaction was stirred at 180 C
for 3 hours and then cooled to room temperature. Ethanol was added; a precipitate formed and was collected by filtration, dried in vacuo to provide intermediate 9-d as a beige solid.
Synthesis of intermediate 10-a Br Ph3P, DIAD
9-d ______________________________ , NH2 .
OH
6 N 1 \
' N N
10-a o Scheme 10 To a solution of intermediate 9-d (1.0 g, 3.45 mmol) in THF was added triphenylphosphine (1.35 g, 5.17 mmol), cyclopentanol (0.47 ml, 5.17 mmol) and DIAD (1.0 ml, 5.17 mmol) and the reaction was then stirred at room temperature overnight. Volatiles were removed under reduced pressure.
Purification by silica gel chromatography provided intermediate 10-a as white solid. MS (m/z) M+H= 359.6 Synthesis of Compound 9 0 a 0 OH imidazole 0 OTBS Ph3P, DIAD, I. 0 w ____________________________________________ 1.
TBSCI CI
11-a 11-b to 11-b _____ ).
OH
11-c 0 Os cuci, CS2CO3, 10-a + 11-c > OH NH2 441k glik CI
lel N N
Compound 9 a Scheme 11 Step 1: Intermediate 11-a To a solution of resorcinol (15.0 g, 136 mmol) in DMF (100 ml), cooled to 0 C, were added imidazole (19.48 g, 286 mmol) and tert-butylchlorodimethylsilane (21.56 g, 143 mmol). The reaction was then stirred at room temperature overnight. A saturated aqueous solution of ammonium chloride and ethyl acetate were added; the organic layer was separated, washed 3 times with a saturated aqueous solution of ammonium chloride and brine, dried over MgSO4, filtered and concentrated under reduced pressure.
Purification by silica gel chromatography provided intermediate 11-a as a colorless oil.
Step 2: Intermediate 11-b To a solution of (4-chlorophenyl) methanol (1.52 g, 10.70 mmol) in THF (20 mL) were sequentially added intermediate 11-a (2.88 g, 12.84 mmol), triphenylphosphine (3.37 g, 12.84 mmol) and DIAD (2.53 ml, 12.84 mmol) drop wise at room temperature and the reaction was then stirred for 1 hour.
A saturated aqueous solution of ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 11-b as a colorless oil.
Step 3: Intermediate 11-c Tetrabutylammonium fluoride trihydrate (3.93 g, 12.47 mmol) was added to a solution of intermediate 11-b (2.9 g, 8.31 mmol) in THF (15 mL) and the reaction was stirred at room temperature overnight. A saturated aqueous solution of ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 11-c as a colorless oil.
Step 4: Compound 9 A solution of intermediate 10-a (200 mg, 0.56 mmol), intermediate 11-c (229 mg, 0.977 mmol), quinolin-8-ol (16.21 mg, 0.112 mmol), copper (I) chloride (11.05 mg, 0.11 mmol) and cesium carbonate (546 mg, 1.67 mmol), in dimethylacetamide (1 ml), was degassed with argon for 10 minutes, heated in a sealed tube at 140 C overnight and then cooled to room temperature. Water and ethyl acetate were added, the organic layer was separated, the aqueous layer was extracted twice with ethyl acetate, the combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% HCl/methanol gradient provided compound 9.HCI as a yellow solid. MS (m/z) M+H= 512.2 Synthesis of intermediate 12-a Br Ph3P, DIAD , 9-d \,N
Me0H N' \
Scheme 12 To a solution of intermediate 9-d (500 mg, 1.72 mmol) in THF (8.6 mL), were sequentially added methanol (105 pl, 2.59 mmol), triphenylphosphine (678 mg, 2.59 mmol) and DIAD (503 pl, 2.59 mmol) drop wise at room temperature. The solution was then stirred at room temperature overnight.
A precipitate formed and was collected by filtration, dried in vacuo to provide intermediate 12-a as a white solid.
Synthesis of compound 16 O, 12-a + 3-a CuCI, CS2CO3, NH2 4Ik OH ___________________________________ N
I ,N
N N
./
Compound 16 Scheme 13 A solution of intermediate 12-a (235 mg, 0.77 mmol), intermediate 3-a (271 mg, 1.35 mmol), quinolin-8-ol (22.4 mg, 0.15 mmol), copper (I) chloride (15.3 mg, 0.15 mmol) and cesium carbonate (755 mg, 2.31 mmol) in dimethylacetamide (1 ml) was degassed with nitrogen for 10 minutes, heated in a sealed tube at 140 C overnight and then cooled to room temperature. Water and ethyl acetate were added, the organic layer was separated, the aqueous layer was extracted twice with ethyl acetate, the combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with 1% HCl/methanol gradient provided compound 16=HCI as a beige solid. MS (m/z) M+H= 424.2 Synthesis of compound 17 is CHO 0 CHO
TEA, TBSCI NaBH4 0 OH
_______________________ v. ______________________ )..-OH OTBS OTBS
14-a 14-b Ph3P, DIAD, 10 0 411 TBAF0 0 i 14-b ________ ). CN l I. CN
OTBS OH
HO S
CN 14-c 14-d 0 . CN
CuCI, CS2CO3, 12-a + 14-d _____________ ).
OH NH2 410 ifk 0 N N ' \N
1 ' N N
\ Compound 17 Scheme 14 Step 1: Intermediate 14-a To a solution of 3-hydroxybenzaldehyde (14.73 g, 121 mmol) in dichloromethane (100 mL) were sequentially added triethylamine (25.08 ml, 181 mmol), tert-butylchlorodimethylsilane (20.0 g, 133 mmol) portion wise, and the reaction was stirred at room temperature overnight. 10% aqueous citric acid was added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure.
Purification by silica gel chromatography provided intermediate 14-a as a yellow oil.
Step 2: Intermediate 14-b To a solution of intermediate 14-a (16.0 g, 67.7 mmol) in methanol (100 ml) cooled to 0 C was added portion wise sodium borohydride (1.28 g, 33.8 mmol). After the addition was completed the reaction was stirred at room temperature for 2 hours. Volatiles were removed under reduced pressure.
Water and ethyl acetate were added to the residue, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure to provide intermediate 14-b as a yellow oil.
Step 3: Intermediate 14-c To a solution of intermediate 14-b (1.0 g, 2.09 mmol) in THF (42 mL) were sequentially added 2-hydroxybenzonitrile (600 mg, 5.03 mmol), triphenylphosphine (1.32 g, 5.03 mmol) and DIAD (991 pl, 5.03 mmol) drop wise at room temperature; the reaction was stirred at reflux for 2 hours and then cooled to room temperature. A saturated aqueous solution of ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 14-c as a colorless oil.
Step 2: Intermediate 14-d To a solution of intermediate 14-c (1.22 g, 3.62 mmol) in THF (36.0 ml) was added tetrabutylammonium fluoride trihydrate (946 mg, 3.62 mmol) and the reaction was stirred at room temperature for 1 hour. A saturated aqueous solution of ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 14-d as a white solid.
Step 2: Compound 17 A solution of intermediate 12-a (200 mg, 0.6 mmol), intermediate 14-d (259 mg, 1.15 mmol), quinolin-8-ol (19.0 mg, 0.13 mmol), copper (I) chloride (13.0 mg, 0.13 mmol) and cesium carbonate (643 mg, 1.97 mmol) in dimethylacetamide (3.0 ml) was degassed with argon for 10 minutes, heated in a sealed tube at 140 C overnight. After cooling to room temperature, water and ethyl acetate were added, the organic layer was separated, the aqueous layer was extracted twice with ethyl acetate, the combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided compound 17 as a white solid. MS (m/z) M+H= 449.3 Synthesis of compound 18:
N
LIAIH4 HOr- ---Ojs----N N Ph3P, DIAD, 0 s 14-b
Representative alkenyl groups include vinyl, propen-2-yl, crotyl, isopenten-2-yl, 1,3-butadien-2-y1), 2,4-pentadienyl, and 1,4-pentadien-3-yl.
Representative alkynyl groups include ethynyl, 1- and 3-propynyl, and 3-butynyl. In certain preferred embodiments, alkyl substituents are lower alkyl groups, e.g., having from 1 to 6 carbon atoms. Similarly, alkenyl and alkynyl preferably refer to lower alkenyl and alkynyl groups, e.g., having from 2 to 6 carbon atoms. As used herein, "alkylene" refers to an alkyl group with two open valencies (rather than a single valency), such as -(Ch12)1-10- and substituted variants thereof.
The term "alkoxy" refers to an alkyl group having an oxygen attached thereto. Representative alkoxy groups include methoxy, ethoxy, propoxy, tert-butoxy and the like. An "ether" is two hydrocarbons covalently linked by an oxygen. Accordingly, the substituent of an alkyl that renders that alkyl an ether is or resembles an alkoxy.
The term "alkoxyalkyl" refers to an alkyl group substituted with an alkoxy group, thereby forming an ether.
The terms "amide" and "amido" are art-recognized as an amino-substituted carbonyl and includes a moiety that can be represented by the general formula:
A ,R10 N
i wherein R9, le) are as defined above. Preferred embodiments of the amide will not include imides, which may be unstable.
The terms "amine" and "amino" are art-recognized and refer to both unsubstituted and substituted amines and salts thereof, e.g., a moiety that can be represented by the general formulae:
1 .,, ¨14 or ¨N4.¨R ''' i \R10 Rio' wherein R9, 111 and R1 ' each independently represent a hydrogen, an alkyl, an alkenyl, -(CH2)m-R8, or R9 and Rw taken together with the N atom to which they are attached complete a heterocycle having from 4 to 8 atoms in the ring structure; R8 represents an aryl, a cycloalkyl, a cycloalkenyl, a heterocyclyl or a polycyclyl; and m is zero or an integer from 1 to 8. In preferred embodiments, only one of R9 or Ftw can be a carbonyl, e.g., R9, R10, and the nitrogen together do not form an imide. In even more preferred embodiments, R9 and le) (and optionally R1w) each independently represent a hydrogen, an alkyl, an alkenyl, or -(CH2)m-R8. In certain embodiments, the amino group is basic, meaning the protonated form has a pKa > 7.00.
The term "aralkyl", as used herein, refers to an alkyl group substituted with an aryl group, for example -(CH2)n-Ar.
The term "heteroaralkyl", as used herein, refers to an alkyl group substituted with a heteroaryl group, for example -(CH2)n-Het.
The term "aryl" as used herein includes 5-, 6-, and 7-membered substituted or unsubstituted single-ring aromatic groups in which each atom of the ring is carbon. The term "aryl" also includes polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjoining rings wherein at least one of the rings is aromatic, e.g., the other cyclic rings can be cycloalkyls, cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and/or heterocyclyls. Aryl groups include benzene, naphthalene, phenanthrene, phenol, aniline, anthracene, and phenanthrene.
The terms "carbocycle" and "carbocyclyl", as used herein, refer to a non-aromatic substituted or unsubstituted ring in which each atom of the ring is carbon. The terms "carbocycle" and "carbocycly1" also include polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjoining rings wherein at least one of the rings is carbocyclic, e.g., the other cyclic rings can be cycloalkyls, cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and/or heterocyclyls. Representative carbocyclic groups include cyclopentyl, cyclohexyl, 1-cyclohexenyl, and 3-cyclohexen-1-yl, cycloheptyl.
The term "carbonyl" is art-recognized and includes such moieties as can be represented by the general formula:
AR"
X' wherein X is a bond or represents an oxygen or a sulfur, and Rn represents a hydrogen, an alkyl, an alkenyl, -(CH2)m-R8 or a pharmaceutically acceptable salt. Where X is an oxygen and R11 is not hydrogen, the formula represents an "ester". Where X is an oxygen, and Rfi is a hydrogen, the formula represents a "carboxylic acid".
The terms "heteroaryl" includes substituted or unsubstituted aromatic 5- to 7-membered ring structures, more preferably 5- to 6-membered rings, whose ring structures include one to four heteroatoms. The term "heteroaryl" also includes polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjoining rings wherein at least one of the rings is heteroaromatic, e.g., the other cyclic rings can be cycloalkyls, cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and/or heterocyclyls. Heteroaryl groups include, for example, pyrrole, furan, thiophene, imidazole, isoxazole, oxazole, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazine and pyrimidine, and the like.
The term "heteroatom" as used herein means an atom of any element other than carbon or hydrogen. Preferred heteroatoms are nitrogen, oxygen, and sulfur.
The terms "heterocycly1" or "heterocyclic group" refer to substituted or unsubstituted non-aromatic 3- to 10-membered ring structures, more preferably 3- to 7-membered rings, whose ring structures include one to four heteroatoms. The term terms "heterocyclyl" or "heterocyclic group" also include polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjoining rings wherein at least one of the rings is heterocyclic, e.g., the other cyclic rings can be cycloalkyls, cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and/or heterocyclyls.
Heterocyclyl groups include, for example, tetrahydrofuran, tetrahydropyran, piperidine, piperazine, pyrrolidine, morpholine, lactones, and lactams.
The term "hydrocarbon", as used herein, refers to a group that is bonded through a carbon atom that does not have a =0 or =S substituent, and typically has at least one carbon-hydrogen bond and a primarily carbon backbone, but may optionally include heteroatoms. Thus, groups like methyl, ethoxyethyl, 2-pyridyl, and trifluoromethyl are considered to be hydrocarbyl for the purposes of this application, but substituents such as acetyl (which has a =0 substituent on the linking carbon) and ethoxy (which is linked through oxygen, not carbon) are not. Hydrocarbyl groups include, but are not limited to aryl, heteroaryl, carbocycle, heterocycle, alkyl, alkenyl, alkynyl, and combinations thereof.
The terms "polycycly1" or "polycyclic" refer to two or more rings (e.g., cycloalkyls, cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and/or heterocyclyls) in which two or more carbons are common to two adjoining rings, e.g., the rings are "fused rings". Each of the rings of the polycycle can be substituted or unsubstituted.
As used herein, the term "probe" means a compound of the invention which is labeled with either a detectable label or an affinity tag, and which is capable of binding, either covalently or non-covalently, to a protein kinase domain. When, for example, the probe is non-covalently bound, it may be displaced by a test compound. When, for example, the probe is bound covalently, it may be used to form cross-linked adducts, which may be quantified and inhibited by a test compound.
The term "substituted" refers to moieties having substituents replacing a hydrogen on one or more carbons of the backbone. It will be understood that "substitution" or "substituted with" includes the implicit proviso that such substitution is in accordance with permitted valence of the substituted atom and the substituent, and that the substitution results in a stable compound, e.g., which does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, etc. As used herein, the term "substituted" is contemplated to include all permissible substituents of organic compounds. In a broad aspect, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of organic compounds. The permissible substituents can be one or more and the same or different for appropriate organic compounds. For purposes of this invention, the heteroatoms such as nitrogen may have hydrogen substituents and/or any permissible substituents of organic compounds described herein which satisfy the valences of the heteroatoms. Substituents can include, for example, a halogen, a hydroxyl, a carbonyl (such as a carboxyl, an alkoxycarbonyl, a formyl, or an acyl), a thiocarbonyl (such as a thioester, a thioacetate, or a thioformate), an alkoxyl, a phosphoryl, a phosphate, a phosphonate, a phosphinate, an amino, an amido, an amidine, an imine, a cyano, a nitro, an azido, a sulfhydryl, an alkylthio, a sulfate, a sulfonate, a sulfamoyl, a sulfonamido, a sulfonyl, a heterocyclyl, an aralkyl, or an aromatic or heteroaromatic moiety. It will be understood by those skilled in the art that the moieties substituted on the hydrocarbon chain can themselves be substituted, if appropriate.
Compounds of the invention also include all isotopes of atoms present in the intermediates and/or final compounds. Isotopes include those atoms having the same atomic number but different mass numbers. For example, isotopes of hydrogen include deuterium and tritium.
General Synthetic Methods General Synthetic Method A:
Ulmann condensation of phenol 1-i with ester 1-ii provided intermediate 1-iii.
Saponification of intermediate 1-iii yielded intermediate 1-iv. Conversion of intermediate 1-iv to its acid chloride, using for example oxalyl chloride and DMF, provided intermediate 1-v. Condensation of intermediate 1-v with malononitrile yielded intermediate 1-vi. Methylation of intermediate 1-vi with TMS-diazomethane provided intermediate 1-vii. Condensation of 1-vii with hydrazine yielded intermediate 1-viii. Condensation of intermediate 1-viii with formamidine yielded intermediate 1-ix. Intermediate 1-ix was treated with alcohol R101-I, under Mitsunobu conditions, to provide the desired compounds or intermediates of general formula 1-x.
(X16. (X16 (X16.
\ ,õ...,,,vv Base, ligand, 1\'-/-w TW
1 catalyst /I- I
..f- NaOH
______________________________________________ ), Y (x2)m OH 0 0 OH
7C) EtO2C (X26 HO2C X2), 1-iii 1-iv 1-ii; X=I, Br (X1),õ. (X1)m.
(x1),,=
t71\1 o W
oxalyl chloride --' malononitrile .vi'm TMS-diazomethane ,j7W
1-iv i ¨(X2)m ' I ¨(X2)rn 1 \
CIOC)(X2)m HOCN Me0 CN
CN CN
1-v 1-vi 1-vii _<------- )m, 0 \ ?
hydrazine / \ W
1-vii _______________________ .
NC -----0(2)m / \,NI
H
1-viii (X1) . (Xl)m 0____.
a m z' c-- \ /
Ph3P, DIAD
W W
.. o. f rmamidine / \
1-viii _______ is. _________________________ 1 NH2 ' (X2)m NH2 ----(X2)m RiOH
\ N N
N ' N I :
vi 1-ix 1-x Scheme 1 General Synthetic Method 13:
Benzoyl chlorides of formula 2-1 were condensed with malononitrile to provide intermediate 2-11. Methylation of intermediate 2-11 with TMS-diazomethane provided intermediate 2-iii. Condensation of intermediate 2-iii with hydrazine provided intermediate 2-iv. Further condensation of intermediate 2-iv with formamidine provided intermediate 2-v. Intermediate 2-v was treated with alcohol R1OH, under Mitsunobu conditions, to provide intermediate 2-vi. Ullmann condensation of intermediate 2-vi with phenolic intermediates 2-vii provided the desired compounds or intermediates of general formula 2-viii.
X
X
X
X
TMS-diazomethane /1-=
.L1 )(2,r, malononitrile rjTh.._(x2) 1 ___(x26 hydrazine ( ) CN o CN NC--.)N
0 Cl HO-1-' CN NH
.'' 2-1; X=1, Br 2-ii; X=1, Br 2-111; X=1, Br 2-iv; X=1, Br X
X / \
formamidine / \ Ph3P, D1AD, 2-iv NH2 -------(X-, )171 IsV , \
1:210H
N \ N N, ' R1 N N
2-v; X=1, Br 2-vi; X=1, Br 0 \ /
Base, ligand, catalyst 2-vi _____________________ p.
NH2 --(X2)m HOW
1 N \
1 ,N
(X1),õ, N N, 2-vii 2-viii Scheme 2a Alternatively, intermediate 2-iv was treated with an alcohol of formula R101-1, under Mitsunobu conditions to provide intermediate 2-ix. Condensation of intermediate 2-ix with formamidine provided intermediate 2-vi.
X
X X
NC Ph3P, DIAD, NC (X26 formamidine NH2 ________________________________________________ k \ RiOH
N N
NH I \ N
H2NN,/
2-iv; X=1, Br 2-ix; X=1, Br 2-vi; X=1, Br Scheme 2b In a similar manner, condensation of intermediate 2-iii with a hydrazine of formula R1NIHNH2 provided intermediate 2-ix. Ullmann condensation of intermediate 2-ix with phenolic intermediates 2-vii provided intermediates 2-x. Condensation of intermediate 2-x with formamidine provided the desired compounds or intermediates of general formula 2-viii.
Xa(xi)m.
, \ 0-7 _(X2),, Base, ligand, \ W
-H1-c2N6 RiNHNH2 NC Ho w . \
NC(X2)m \
CN Ri /'" H HN14 --(X1),,, Ri 2-iii; X=1, Br 2-ix; X=1, Br 2-vii 2-x -(X1)m' 0_ \ /
W
/ \ 2 formamidine 2-x ________________________ .
N \ N
I ' le---N
Ft1 2-viii Scheme 2c Alternatively, trimethyl orthoformate and ammonia can be used in place of formamidine, for example, in the conversion of intermediate 2-x to compounds of formula 2-vii.
Exemplification The following synthetic methods are intended to be representative of the chemistry used to prepare compounds of Formula 1 and are not intended to be limiting.
Synthesis of Compound 1:
0 OBn OH K2CO3 0 OBn CuCI, Cs2CO3 ___________________ r _____________________ ).
Benzyl bromide 0 0 OH OH 40 o'' Ir EtO2C
3-a Br 3-b 0 OBn 0 OBn NaOH oxalyl chloride 3-b __________ r id 0 _________ N ja 0 Mr ir 3-c 3-d o IS o 010 3-d DIPEA 0 TMS-diazomethane 0 malononitrile 0 40 '-.
HO Me0 CN --. CN
CN CN
3-e 3-f 0 * O, hydrazine 41 NC 0 formamidine 3-f r ___________________________ r i \isl . N '' , \ it H2N N, L i ,N
H N N
H
3-g Compound 1 Scheme 3 Step 1: Intermediate 3-a Benzyl bromide (27.0 ml, 227 mmol) was added drop wise to a stirred suspension of resorcinol (25.0 g, 227 mmol) and potassium carbonate (31.4 g, 227 mmol) in acetone (150 ml) and the reaction was heated under reflux overnight. Volatiles were removed under reduced pressure. Water and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure.
Purification by silica gel chromatography provided intermediate 3-a as a beige oil.
Step 2: Intermediate 3-b To a solution of compound 3-a (15.0 g, 74.9 mmol) in 1,4-dioxane (200 ml) were sequentially added ethyl 4-bromobenzoate (20.59 g, 90 mmol), N,N-dimethylglycine (4.25 g, 41.2 mmol), copper(I) chloride(3.71 g, 37.5 mmol) and cesium carbonate (61.0 g, 187 mmol). The reaction mixture was stirred at reflux overnight and then cooled to room temperature. Water and ethyl acetate were added, the organic layer was separated, washed with saturated aqueous NaHCO3, brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 3-b as a colorless oil.
Step 3: Intermediate 3-c To a solution of intermediate 3-b (17.5 g, 50.2 mmol) in THF (200 ml) and Me0H (100 ml) was added 2N sodium hydroxide (100 ml, 200 mmol) and the reaction was stirred at room temperature overnight. Volatiles were removed under reduced pressure. 10% aqueous HCI and ethyl acetate were added to the residue, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure to provide intermediate 3-c as beige solid.
Step 4: Intermediate 3-d To a suspension of intermediate 3-c (16.1 g, 50.3 mmol) in dichloromethane (100 ml) were added DMF (0.1 ml, 1.29 mmol) and oxalyl chloride (4.4 ml, 50.3 mmol). The solution was stirred at room temperature for 2 hours.
Volatiles were removed under reduced pressure to provide intermediate 3-d as beige solid.
Step 5: Intermediate 3-e To a solution of intermediate 3-d (16.5 g, 48.9 mmol) in toluene (50 ml) and THF (7 ml), cooled to -10 C, were added malononitrile (3.19 ml, 50.2 mmol) and DIPEA (17.5 ml, 100 mmol) in toluene (50 mL), drop wise, over a period of 30 minutes. After the addition was completed, the reaction was stirred for 1 hour at 0 C and room temperature overnight. Volatiles were removed under reduced pressure. 1M aqueous HCI and ethyl acetate were added, the organic layer was separated, washed with 1M HCI and brine, dried over MgSO4, filtered and concentrated under reduced pressure to provide intermediate 3-e as beige solid.
Step 6: Intermediate 3-f To a solution of intermediate 3-e (18.1 g, 49.1 mmol) in acetonitrile (177 ml) and methanol (19.0 ml), cooled to 0 C, were added DIPEA (10.3 ml, 59.0 mmol) and a 2M solution of (diazomethyl)trimethylsilane in hexanes (27.0 ml, 54.0 mmol). After the addition was completed, the reaction was stirred at room temperature overnight. Acetic acid (0.56 ml, 9.83 mmol) was added, the reaction was then stirred for 30 minutes and volatiles were removed under reduced pressure. A saturated aqueous solution of NaHCO3 and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure.
Purification by silica gel chromatography provided intermediate 3-f as yellow solid.
Step 7: Intermediate 3-g To a suspension of intermediate 3-f (8.05 g, 21.1 mmol) in ethanol (10.5 ml) was added a solution of hydrazine monohydrate (2.76 ml, 56.8 mmol). The reaction was stirred at 100 C for 1 hour and then cooled to room temperature.= Water was added; a precipitate formed and was collected by filtration, washed with diethyl ether and dried in vacuo to provide intermediate 3-g as an off-white solid.
Step 8: Compound 1 Intermediate 3-g (8.0 g, 20.92 mmol) was added to a solution of formamidine (58.4 ml, 1464 mmol) and the reaction was stirred at 180 C
for 2 hours and then cooled to room temperature. Water and ethyl acetate were added; the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure to provide compound 1 as beige solid. MS (m/z) M+H=410.2 Synthesis of Compound 2:
O . o 40 .
o o NH2 41k 40 Ph3P, DIAD NH2 N' \N N
OH N
i N' [.., 1 )1 N
H
uCompound 1 Compound 2 Scheme 4 To a solution of cyclopentanol (316 mg, 3.66 mmol) in THF was added triphenylphosphine (961 mg, 3.66 mmol) and DIAD (712 pl, 3.66 mmol). The yellow solution was stirred for 5 minutes, compound 1 (1.0 g, 2.44 mmol) was added and the reaction was then stirred at room temperature overnight.
Volatiles were removed under reduced pressure. Purification by silica gel chromatography provided compound 2 as an off-white solid. MS (m/z) M+H=478.2 Synthesis of Compound 3:
, o 0 NH2 * Ph3P, DIAD
N
N 41#
N
' N
N N' Ny(31 N N
Compound 1 Compound 3 Scheme 5 To a solution of (S)-tert-butyl 3-hydroxypiperidine-1-carboxylate (5.65 g, 28.1 mmol) in THF was added triphenylphosphine (7.37 g, 28.1 mmol) and DIAD (5.46 ml, 28.1 mmol). The yellow solution was stirred for 5 minutes, compound 1 (10.0 g, 24.42 mmol) was added and the reaction was then stirred at room temperature overnight. Volatiles were removed under reduced pressure. Purification by silica gel chromatography provided compound 3 as a white foam. MS (m/z) M+H= 593.1 Synthesis of Compound 4:
o NH2 41, HCI
NH2 *
\ N
I N I ,N
N N N
LNH
Compound 4 Compound 3 Scheme 6 To a solution of compound 3 (1.88 g, 3.17 mmol) in dichloromethane was added 4N HCI in 1,4-dioxane (19.82 ml, 79.0 mmol) and the reaction was stirred at room temperature for 2 hours. Volatiles were removed under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 4.2HCI as a white solid. MS (m/z) M+H= 493.1 Synthesis of compound 5 0 = o, TEA
4Ik 0 NH2 N \
'N I N
òH
Compound 4 Compound 5 Scheme 7 To a solution of compound 4.2HCI (100 mg, 0.17 mmol) in dichloromethane (2 ml) cooled to 0 C were sequentially added TEA (99 pl, 0.70 mmol) and acryloyl chloride (17.6 mg, 0.19 mmol). The reaction was stirred at 0 C for 1 hour. A saturated aqueous solution of ammonium chloride was added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided compound 5 as a white solid. MS (m/z) M+H=
547.1 Synthesis of compound 6 ifh o, TEA
0 =
41kN \ 11 CI N \ N
I , N'N N
L\NH LN
Compound 4 Compound 6 Scheme 8 To a solution of compound 4.2HCI (1.8 g, 3.18 mmol) in dichloromethane (32 ml) cooled to 0 C were sequentially added TEA (1.77 ml, 12.73 mmol) and acetyl chloride (249 pl, 3.50 mmol). The reaction was stirred at 0 C for 1 hour and room temperature overnight. A saturated aqueous solution of ammonium chloride was added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with 1%
aqueous HCl/methanol gradient provided compound 6.11C1 as beige solid. MS
(m/z) M+H= 535.1 Synthesis of intermediate 9-d Br Br Br 0 DIPEA , malononitrile (110 TMS-diazomethane SI
________________________________________________ A
CN =-. \ CN
HO
0 CI , CN
CN
9-a 9-b Br Br hydrazine 0 formamidine 9-b __________ A ' NH2 fa NC \'N N \ N
NH N N
H
9-c 9-d Scheme 9 Step 1: Intermediate 9-a To a solution of 4-bromobenzoyl chloride (25.0 g, 114 mmol) in toluene (200 ml) and THF (30 ml), cooled to -10 C, were sequentially added malononitrile (7.60 ml, 120.0 mmol) and DIPEA (39.8 ml, 228 mmol) in toluene (50 mL) drop wise over a period of 1 hour. After the addition was completed, the reaction was stirred for 1 hour at 0 C and room temperature overnight.
Volatiles were removed under reduced pressure. 1N HCI and ethyl acetate were added to the residue, the organic layer was separated, washed twice with 1N HCI and brine, dried over MgSO4, filtered and concentrated under reduced pressure to provide intermediate 9-a as yellow solid.
Step 2: Intermediate 9-b To a solution of intermediate 9-a (26.4 g, 106 mmol) in acetonitrile (300 ml) and methanol (35.0 ml), cooled to 0 C, was added DIPEA (22.2 ml, 127 mmol) and a 2M solution of diazomethyl)trimethylsilane in hexanes (58.3 ml, 117 mmol). After the addition was completed, the reaction was stirred at room temperature overnight. Acetic acid (1.21 ml, 21.2 mmol) was added, the reaction was stirred for 30 minutes and volatiles were removed under reduced pressure. A saturated aqueous solution of NaHCO3 and ethyl acetate were added, the organic layer was separated, washed with brine, dried over M9SO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 9-b as a yellow solid.
Step 3: Intermediate 9-c To a suspension of intermediate 9-b (4.49 g, 17.07 mmol) in ethanol (8.5 ml) was added a solution of hydrazine monohydrate (2.23 ml, 46.1 mmol) and the reaction was stirred at 100 C for 1 hour and then cooled to room temperature. Volatiles were removed under reduced pressure to provide intermediate 9-c as a yellow solid.
Step 4: Intermediate 9-d Intermediate 9-c (4.49 g, 17.07 mmol) was added to a solution of formamidine (40.8 ml, 1024 mmol) and the reaction was stirred at 180 C
for 3 hours and then cooled to room temperature. Ethanol was added; a precipitate formed and was collected by filtration, dried in vacuo to provide intermediate 9-d as a beige solid.
Synthesis of intermediate 10-a Br Ph3P, DIAD
9-d ______________________________ , NH2 .
OH
6 N 1 \
' N N
10-a o Scheme 10 To a solution of intermediate 9-d (1.0 g, 3.45 mmol) in THF was added triphenylphosphine (1.35 g, 5.17 mmol), cyclopentanol (0.47 ml, 5.17 mmol) and DIAD (1.0 ml, 5.17 mmol) and the reaction was then stirred at room temperature overnight. Volatiles were removed under reduced pressure.
Purification by silica gel chromatography provided intermediate 10-a as white solid. MS (m/z) M+H= 359.6 Synthesis of Compound 9 0 a 0 OH imidazole 0 OTBS Ph3P, DIAD, I. 0 w ____________________________________________ 1.
TBSCI CI
11-a 11-b to 11-b _____ ).
OH
11-c 0 Os cuci, CS2CO3, 10-a + 11-c > OH NH2 441k glik CI
lel N N
Compound 9 a Scheme 11 Step 1: Intermediate 11-a To a solution of resorcinol (15.0 g, 136 mmol) in DMF (100 ml), cooled to 0 C, were added imidazole (19.48 g, 286 mmol) and tert-butylchlorodimethylsilane (21.56 g, 143 mmol). The reaction was then stirred at room temperature overnight. A saturated aqueous solution of ammonium chloride and ethyl acetate were added; the organic layer was separated, washed 3 times with a saturated aqueous solution of ammonium chloride and brine, dried over MgSO4, filtered and concentrated under reduced pressure.
Purification by silica gel chromatography provided intermediate 11-a as a colorless oil.
Step 2: Intermediate 11-b To a solution of (4-chlorophenyl) methanol (1.52 g, 10.70 mmol) in THF (20 mL) were sequentially added intermediate 11-a (2.88 g, 12.84 mmol), triphenylphosphine (3.37 g, 12.84 mmol) and DIAD (2.53 ml, 12.84 mmol) drop wise at room temperature and the reaction was then stirred for 1 hour.
A saturated aqueous solution of ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 11-b as a colorless oil.
Step 3: Intermediate 11-c Tetrabutylammonium fluoride trihydrate (3.93 g, 12.47 mmol) was added to a solution of intermediate 11-b (2.9 g, 8.31 mmol) in THF (15 mL) and the reaction was stirred at room temperature overnight. A saturated aqueous solution of ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 11-c as a colorless oil.
Step 4: Compound 9 A solution of intermediate 10-a (200 mg, 0.56 mmol), intermediate 11-c (229 mg, 0.977 mmol), quinolin-8-ol (16.21 mg, 0.112 mmol), copper (I) chloride (11.05 mg, 0.11 mmol) and cesium carbonate (546 mg, 1.67 mmol), in dimethylacetamide (1 ml), was degassed with argon for 10 minutes, heated in a sealed tube at 140 C overnight and then cooled to room temperature. Water and ethyl acetate were added, the organic layer was separated, the aqueous layer was extracted twice with ethyl acetate, the combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% HCl/methanol gradient provided compound 9.HCI as a yellow solid. MS (m/z) M+H= 512.2 Synthesis of intermediate 12-a Br Ph3P, DIAD , 9-d \,N
Me0H N' \
Scheme 12 To a solution of intermediate 9-d (500 mg, 1.72 mmol) in THF (8.6 mL), were sequentially added methanol (105 pl, 2.59 mmol), triphenylphosphine (678 mg, 2.59 mmol) and DIAD (503 pl, 2.59 mmol) drop wise at room temperature. The solution was then stirred at room temperature overnight.
A precipitate formed and was collected by filtration, dried in vacuo to provide intermediate 12-a as a white solid.
Synthesis of compound 16 O, 12-a + 3-a CuCI, CS2CO3, NH2 4Ik OH ___________________________________ N
I ,N
N N
./
Compound 16 Scheme 13 A solution of intermediate 12-a (235 mg, 0.77 mmol), intermediate 3-a (271 mg, 1.35 mmol), quinolin-8-ol (22.4 mg, 0.15 mmol), copper (I) chloride (15.3 mg, 0.15 mmol) and cesium carbonate (755 mg, 2.31 mmol) in dimethylacetamide (1 ml) was degassed with nitrogen for 10 minutes, heated in a sealed tube at 140 C overnight and then cooled to room temperature. Water and ethyl acetate were added, the organic layer was separated, the aqueous layer was extracted twice with ethyl acetate, the combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with 1% HCl/methanol gradient provided compound 16=HCI as a beige solid. MS (m/z) M+H= 424.2 Synthesis of compound 17 is CHO 0 CHO
TEA, TBSCI NaBH4 0 OH
_______________________ v. ______________________ )..-OH OTBS OTBS
14-a 14-b Ph3P, DIAD, 10 0 411 TBAF0 0 i 14-b ________ ). CN l I. CN
OTBS OH
HO S
CN 14-c 14-d 0 . CN
CuCI, CS2CO3, 12-a + 14-d _____________ ).
OH NH2 410 ifk 0 N N ' \N
1 ' N N
\ Compound 17 Scheme 14 Step 1: Intermediate 14-a To a solution of 3-hydroxybenzaldehyde (14.73 g, 121 mmol) in dichloromethane (100 mL) were sequentially added triethylamine (25.08 ml, 181 mmol), tert-butylchlorodimethylsilane (20.0 g, 133 mmol) portion wise, and the reaction was stirred at room temperature overnight. 10% aqueous citric acid was added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure.
Purification by silica gel chromatography provided intermediate 14-a as a yellow oil.
Step 2: Intermediate 14-b To a solution of intermediate 14-a (16.0 g, 67.7 mmol) in methanol (100 ml) cooled to 0 C was added portion wise sodium borohydride (1.28 g, 33.8 mmol). After the addition was completed the reaction was stirred at room temperature for 2 hours. Volatiles were removed under reduced pressure.
Water and ethyl acetate were added to the residue, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure to provide intermediate 14-b as a yellow oil.
Step 3: Intermediate 14-c To a solution of intermediate 14-b (1.0 g, 2.09 mmol) in THF (42 mL) were sequentially added 2-hydroxybenzonitrile (600 mg, 5.03 mmol), triphenylphosphine (1.32 g, 5.03 mmol) and DIAD (991 pl, 5.03 mmol) drop wise at room temperature; the reaction was stirred at reflux for 2 hours and then cooled to room temperature. A saturated aqueous solution of ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 14-c as a colorless oil.
Step 2: Intermediate 14-d To a solution of intermediate 14-c (1.22 g, 3.62 mmol) in THF (36.0 ml) was added tetrabutylammonium fluoride trihydrate (946 mg, 3.62 mmol) and the reaction was stirred at room temperature for 1 hour. A saturated aqueous solution of ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 14-d as a white solid.
Step 2: Compound 17 A solution of intermediate 12-a (200 mg, 0.6 mmol), intermediate 14-d (259 mg, 1.15 mmol), quinolin-8-ol (19.0 mg, 0.13 mmol), copper (I) chloride (13.0 mg, 0.13 mmol) and cesium carbonate (643 mg, 1.97 mmol) in dimethylacetamide (3.0 ml) was degassed with argon for 10 minutes, heated in a sealed tube at 140 C overnight. After cooling to room temperature, water and ethyl acetate were added, the organic layer was separated, the aqueous layer was extracted twice with ethyl acetate, the combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided compound 17 as a white solid. MS (m/z) M+H= 449.3 Synthesis of compound 18:
N
LIAIH4 HOr- ---Ojs----N N Ph3P, DIAD, 0 s 14-b
15-a 15-b N
----TBAF
S
15-b OH
15-c O, CuCI, CS2CO3 4.0---\-12-a +15-c ON NH2 S/, N
\
N
IW N -- 1 1 \
L ,N
/
N N
\ Compound 18 Scheme 15 Step 1: Intermediate 15-a To a solution of ethyl 2-methylthiazole-5-carboxylate (5.82 g, 34.0 mmol) in THF (170 m1), cooled to 0 C, was added a 1.0M solution of LiAIH4in THF
(34.0 ml, 34.0 mmol) and the reaction was slowly warmed to room temperature and stirred overnight. Water (1.3 ml) was slowly added, followed by 15% NaOH (1.3 mL). The solution was stirred for 2 hours at room temperature then filtered over celite. The filtrate was concentrated under reduced pressure to provide intermediate 15-a as a yellow oil.
Step 2: Intermediate 15-b To a solution of intermediate 15-a (7.75 g, 34.5 mmol) and intermediate 11-a (4.25 g, 32.9 mmol), in THF (33 mL), were sequentially added triphenylphosphine (10.35 g, 39.5 mmol) and DIAD (7.68 ml, 39.5 mmol) drop wise at room temperature. The reaction was then stirred for 18 hours.
Volatiles were removed in vacuo. Purification by silica gel chromatography provided intermediate 15-b as a colorless oil.
Step 3: Intermediate 15-c To a solution of intermediate 15-b (5.5 g, 16.39 mmol), in THF (82.0 ml), was added a 1.0M solution of tetrabutylammonium fluoride in THF (16.4 ml,
----TBAF
S
15-b OH
15-c O, CuCI, CS2CO3 4.0---\-12-a +15-c ON NH2 S/, N
\
N
IW N -- 1 1 \
L ,N
/
N N
\ Compound 18 Scheme 15 Step 1: Intermediate 15-a To a solution of ethyl 2-methylthiazole-5-carboxylate (5.82 g, 34.0 mmol) in THF (170 m1), cooled to 0 C, was added a 1.0M solution of LiAIH4in THF
(34.0 ml, 34.0 mmol) and the reaction was slowly warmed to room temperature and stirred overnight. Water (1.3 ml) was slowly added, followed by 15% NaOH (1.3 mL). The solution was stirred for 2 hours at room temperature then filtered over celite. The filtrate was concentrated under reduced pressure to provide intermediate 15-a as a yellow oil.
Step 2: Intermediate 15-b To a solution of intermediate 15-a (7.75 g, 34.5 mmol) and intermediate 11-a (4.25 g, 32.9 mmol), in THF (33 mL), were sequentially added triphenylphosphine (10.35 g, 39.5 mmol) and DIAD (7.68 ml, 39.5 mmol) drop wise at room temperature. The reaction was then stirred for 18 hours.
Volatiles were removed in vacuo. Purification by silica gel chromatography provided intermediate 15-b as a colorless oil.
Step 3: Intermediate 15-c To a solution of intermediate 15-b (5.5 g, 16.39 mmol), in THF (82.0 ml), was added a 1.0M solution of tetrabutylammonium fluoride in THF (16.4 ml,
16.4 mmol) and the reaction was stirred at room temperature for 30 minutes. A saturated aqueous solution of ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgS0.4, filtered and concentrated under reduced pressure.
Purification by silica gel chromatography provided intermediate 15-c as beige solid.
Step 4: Compound 18 A solution of intermediate 12-a (200 mg, 0.65 mmol), intermediate 15-c (146 mg, 0.65 mmol), quinolin-8-ol (19.0 mg, 0.13 mmol), copper (I) chloride (13.0 mg, 0.13 mmol) and cesium carbonate (643 mg, 1.97 mmol) in dimethylacetamide (6.5 ml) was degassed with argon for 10 minutes, heated in a sealed tube at 140 C for 2 hours and then cooled to room temperature. Water and ethyl acetate were added, the organic layer was separated, the aqueous layer was extracted twice with ethyl acetate, the combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with 1% HCl/methanol gradient provided compound 18=2HCI as a beige solid. MS (m/z) M+H= 445.1 Synthesis of compound 15:
0 ilikt 0 CN
CuCI, Cs2CO3 10-a + 14-d __________________ v, NH2 O OOH
N N.' \
I t NN el N - Compound 15 a Scheme 16 A solution of intermediate 10-a (200 mg, 0.56 mmol), intermediate 14-d (156 mg, 0.68 mmol), quinolin-8-ol (16.2 mg, 0.11 mmol), copper (I) chloride (11.0 mg, 0.11 mmol) and cesium carbonate (546 mg, 1.67 mmol) in dimethylacetamide (5.5 ml) was degassed with argon for 10 minutes, heated in a sealed tube at 140 C overnight and then cooled to room temperature. Water and ethyl acetate were added, the organic layer was separated, the aqueous layer was extracted twice with ethyl acetate, the combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with 1% HCl/methanol gradient provided compound 15=HCI as beige solid. MS (m/z) M+H= 503.3 Synthesis of intermediate 17-a Br NH2 .
Ph3P, DIAD
9-d N *' \
__________________________________ A I ,N
0/ )¨OH N N)Th \_
Purification by silica gel chromatography provided intermediate 15-c as beige solid.
Step 4: Compound 18 A solution of intermediate 12-a (200 mg, 0.65 mmol), intermediate 15-c (146 mg, 0.65 mmol), quinolin-8-ol (19.0 mg, 0.13 mmol), copper (I) chloride (13.0 mg, 0.13 mmol) and cesium carbonate (643 mg, 1.97 mmol) in dimethylacetamide (6.5 ml) was degassed with argon for 10 minutes, heated in a sealed tube at 140 C for 2 hours and then cooled to room temperature. Water and ethyl acetate were added, the organic layer was separated, the aqueous layer was extracted twice with ethyl acetate, the combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with 1% HCl/methanol gradient provided compound 18=2HCI as a beige solid. MS (m/z) M+H= 445.1 Synthesis of compound 15:
0 ilikt 0 CN
CuCI, Cs2CO3 10-a + 14-d __________________ v, NH2 O OOH
N N.' \
I t NN el N - Compound 15 a Scheme 16 A solution of intermediate 10-a (200 mg, 0.56 mmol), intermediate 14-d (156 mg, 0.68 mmol), quinolin-8-ol (16.2 mg, 0.11 mmol), copper (I) chloride (11.0 mg, 0.11 mmol) and cesium carbonate (546 mg, 1.67 mmol) in dimethylacetamide (5.5 ml) was degassed with argon for 10 minutes, heated in a sealed tube at 140 C overnight and then cooled to room temperature. Water and ethyl acetate were added, the organic layer was separated, the aqueous layer was extracted twice with ethyl acetate, the combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with 1% HCl/methanol gradient provided compound 15=HCI as beige solid. MS (m/z) M+H= 503.3 Synthesis of intermediate 17-a Br NH2 .
Ph3P, DIAD
9-d N *' \
__________________________________ A I ,N
0/ )¨OH N N)Th \_
17-a U0 Scheme 17 To a solution of intermediate 9-d (650 mg, 2.24 mmol), in THF (22.0 mL), were sequentially added tetrahydro-2H-pyran-4-ol (320 pl, 3.36 mmol), triphenylphosphine (881 mg, 3.36 mmol) and DIAD (653 pl, 3.36 mmol) drop wise at room temperature. The solution was then stirred at 50 C
overnight. Volatiles were removed in vacuo. Purification by silica gel chromatography provided intermediate 17-a as a white solid.
Synthesis of compound 22:
0 . CN
CuCI, Cs2CO3 17-a + 14-d OH __ ). NH2 44k 4411k N
=NV "
i 1 ,N
N N Compound 22 a Scheme 18 A solution of intermediate 17-a (200 mg, 0.53mmol), intermediate 14-d (181 mg, 0.80 mmol), quinolin-8-ol (15.5 mg, 0.11 mmol), copper (I) chloride (11.5 mg, 0.11 mmol) and cesium carbonate (348 mg, 1.07 mmol) in dimethylacetamide (5.3 ml) was degassed with argon for 10 minutes, heated in a sealed tube at 140 C overnight and then cooled to room temperature.
Water and ethyl acetate were added, the organic layer was separated, the aqueous layer was extracted twice with ethyl acetate, the combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with 1% HCl/methanol gradient provided compound 22=HCI as a beige solid. MS (m/z) M+H= 519.2 Synthesis of compound 31:
Ph3P, DIAD, o TBAF 0 1.1 14-b CF3 OTBS OH
HO SI
CF3 19-a 19-b CuCI, Cs2CO3, 0 12-a + 19-b N \N
I ' / N N ¨
\ Compound 31 Scheme 19 Step 1: Intermediate 19-a To a solution of intermediate 14-b (10.0 g, 41.9 mmol) in THF (210 mL) were sequentially added 2-(trifluoromethyl)phenol (6.80 g, 41.9 mmol), triphenylphosphine (13.2 g, 50.33 mmol) and DIAD (9.79 ml, 50.3 mmol) drop wise at room temperature. The reaction was then stirred at room temperature overnight. Saturated aqueous ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure.
Purification by silica gel chromatography provided intermediate 19-a as a colorless oil.
Step 2: Intermediate 19-b To a solution of intermediate 19-a (13.9 g, 36.3 mmol) in THF (182.0 ml) was added a 1.0M solution of tetrabutylammonium fluoride in THF (36.3 ml, 36.3 mmol) and the reaction was stirred at room temperature for 1 hour.
Saturated aqueous ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 19-b as colorless oil.
Step 3: Compound 31 A solution of intermediate 12-a (200 mg, 0.66 mmol), intermediate 19-b (265 mg, 0.98 mmol), quinolin-8-ol (19.0 mg, 0.13 mmol), copper (I) chloride (25.5 mg, 0.13 mmol) and cesium carbonate (429 mg, 1.31 mmol) in dimethylacetamide (6.5 ml) was degassed with argon for 10 minutes, heated in a sealed tube at 140 C overnight and then cooled to room temperature. Water and ethyl acetate were added, the organic layer was separated, the aqueous layer was extracted twice with ethyl acetate, the combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with 1% HCl/methanol gradient provided compound 31.HCI as white solid. MS (m/z) M+H= 492.1 Synthesis of compound 32:
0 41, o 0F3 Cul, Cs2CO3 10-a + 19-b OH __ lw- NH2 ft 441k N N -' \
I 'N
4101 N N Compound 32 a Scheme 20 A solution of intermediate 10-a (200 mg, 0.55 mmol), intermediate 19-b (225 mg, 0.83 mmol), quinolin-8-ol (16.2 mg, 0.11 mmol), copper (I) iodide (22.0 mg, 0.11 mmol) and cesium carbonate (364 mg, 1.17 mmol) in dimethylacetamide (5.5 ml) was degassed with argon for 10 minutes, heated in a sealed tube at 140 C overnight and then cooled to room temperature.
Water and ethyl acetate were added, the organic layer was separated, the aqueous layer was extracted twice with ethyl acetate. The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with 1% HCl/methanol gradient provided compound 32.HCI as beige solid. MS (m/z) M+H= 546.1 Synthesis of compound 36:
0 41kt u3 cul, Cs2CO3 17-a + 19-b OH __ 11. NH2 0 O
' N N Compound 36 1 / o Scheme 21 A solution of intermediate 17-a (200 mg, 0.53 mmol), intermediate 19-b (215 mg, 0.80 mmol), quinolin-8-ol (15.5 mg, 0.11 mmol), copper (I) iodide (20.3 mg, 0.11 mmol) and cesium carbonate (348 mg, 1.06 mmol) in dimethylacetamide (5.3 ml) was degassed with argon for 10 minutes, heated in a sealed tube at 140 C overnight and then cooled to room temperature.
Water and ethyl acetate were added, the organic layer was separated, the aqueous layer was extracted twice with ethyl acetate. The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Puriflcation by reverse phase chromatography eluting with 1% HCl/methanol gradient provided compound 36=HCI as beige solid. MS (m/z) M+H= 562.2 Synthesis of compound 20:
to 0 Ph3P, DIAD, 0 TBAF 0 HO __________________ ).. i.
OTBS
CF3 las OTBS OH
11-a 22-b HO 22-a 0 . CF3 CuCI, Cs2CO3, 12-a + 22-b ' NH
N N \
i I N
' 101 N N
\ Compound 20 Scheme 22 Step 1: Intermediate 22-a To a solution of 2-(trifluoromethyl)phenylmethanol (1.43 g, 8.10 mmol) in THF (8.10 mL) were sequentially added intermediate 11-a (2.0 g, 8.91 mmol), triphenylphosphine (2.55 g, 9.72 mmol) and DIAD (1.89 ml, 9.72 mmol) drop wise at room temperature. The reaction was then stirred overnight at room temperature. Saturated aqueous ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure.
Purification by silica gel chromatography provided intermediate 22-a as a colorless oil.
Step 2: Intermediate 22-b Tetrabutylammonium fluoride trihydrate (1.81 g, 5.75 mmol) was added to a solution of intermediate 22-a (2.2 g, 5.75 mmol) in THF (23 mL) and the reaction was stirred at room temperature for 1 hour. Saturated aqueous ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 22-b as a colorless oil.
Step 3: Compound 20 A solution of intermediate 12-a (200 mg, 0.65 mmol), intermediate 22-b (309 mg, 1.15 mmol), quinolin-8-ol (19.1 mg, 0.13 mmol), copper (I) chloride (13.0 mg, 0.13 mmol) and cesium carbonate (429 mg, 1.31 mmol), in dimethylacetamide (6.5 ml), was degassed with argon for 10 minutes, heated in a sealed tube at 140 C overnight and then cooled to room temperature. Water and ethyl acetate were added, the organic layer was separated, the aqueous layer was extracted twice with ethyl acetate, the combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% HCl/methanol gradient provided compound 20.1-1CI as beige solid. MS (m/z) M+H= 492.1 Synthesis of compound 29:
0 =
cF3 Cul, Cs2CO3 10-a + 22-b OH __ 1.- NH2 O 4411k N N'' \N
I
0 N N Compound 29 a Scheme 23 A solution of intermediate 10-a (200 mg, 0.55 mmol), intermediate 22-b (225 mg, 0.83 mmol), quinolin-8-ol (16.2 mg, 0.11 mmol), copper (I) iodide (21.2 mg, 0.11 mmol) and cesium carbonate (364 mg, 1.11 mmol), in dimethylacetamide (5.5 ml), was degassed with argon for 10 minutes, heated in a sealed tube at 140 C overnight and then cooled to room temperature. Water and ethyl acetate were added, the organic layer was separated, the aqueous layer was extracted twice with ethyl acetate, the combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% HCl/methanol gradient provided compound 29=HCI as beige solid. MS (m/z) M+H= 546.2 Synthesis of compound 23:
0 =
cF3 Cul, Cs2CO3 17-a + 22-b OH __ ii. NH2 0 40 N N \N
I N
Ol / N _.....) Compound Scheme 24 A solution of intermediate 17-a (200 mg, 0.53 mmol), intermediate 22-b (215 mg, 0.80 mmol), quinolin-8-ol (15.5 mg, 0.11 mmol), copper (I) iodide (20.4 mg, 0.11 mmol) and cesium carbonate (348 mg, 1.07 mmol), in dimethylacetamide (5.3 ml), was degassed with argon for 10 minutes, heated in a sealed tube at 140 C overnight and then cooled to room temperature. Water and ethyl acetate were added, the organic layer was separated, the aqueous layer was extracted twice with ethyl acetate, the combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% HCl/methanol gradient provided compound 23.1-1CI as beige solid. MS (m/z) M+H= 562.1 Synthesis of compound 30:
NC
=
OH
K2CO3 10 =
=
OH Br 4101 OH
CN 25-a CN
Cul, Cs2CO3, 12-a + 25-a ________________ = OH NH2 4Ik \
N
Compound 30 Scheme 25 Step 1: Intermediate 25-a To a solution of 2-(bromomethypbenzonitrile (1.0 g, 5.10 mmol) and resorcinol (2.81 g, 25.5 mmol) in acetone (51.0 mL) was added cesium carbonate (3.32 g, 10.20 mmol) and the reaction was then stirred at reflux for 2 hours. Volatiles were removed under reduced pressure. Saturated aqueous ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 25-a as white solid.
Step 2: Compound 30 A solution of intermediate 12-a (200 mg, 0.65 mmol), intermediate 25-a (222 mg, 0.98 mmol), quinolin-8-ol (19.1 mg, 0.13 mmol), copper (I) iodide (25.0 mg, 0.13 mmol) and cesium carbonate (429 mg, 1.31 mmol), in dimethylacetamide (6.5 ml), was degassed with argon for 10 minutes, heated in a sealed tube at 140 C overnight and then cooled to room temperature. Water and ethyl acetate were added, the organic layer was separated, the aqueous layer was extracted twice with ethyl acetate, the combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% HCl/methanol gradient provided compound 30=HCI as beige solid. MS (m/z) M+H= 449.4 Synthesis of compound 12:
CuCI, Cs2CO3 0 10-a + 25-a ___________________ s NH2 fa O
OH
N N
I 1' .1 .' N N - Compound 12 a Scheme 26 A solution of intermediate 10-a (200 mg, 0.55 mmol), intermediate 25-a (220 mg, 0.98 mmol), quinolin-8-ol (16.2 mg, 0.11 mmol), copper (I) chloride (11.0 mg, 0.11 mmol) and cesium carbonate (546 mg, 1.67 mmol), in dimethylacetamide (5.5 ml), was degassed with argon for 10 minutes, heated in a sealed tube at 140 C overnight and then cooled to room temperature. Water and ethyl acetate were added, the organic layer was separated, the aqueous layer was extracted twice with ethyl acetate, the combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% HCl/methanol gradient provided compound 12=HCI as a beige solid. MS (m/z) M+H= 503.2 Synthesis of compound 35:
0 . CN
Cul, Cs2CO3 0 17-a + 25-a __________________ D.- NH2 . 0 OH
N I N.' \N ' 11101 .7N
N ¨ Compound 35 o Scheme 27 A solution of intermediate 17-a (200 mg, 0.55 mmol), intermediate 25-a (181.0 mg, 0.80 mmol), quinolin-8-ol (15.5 mg, 0.11 mmol), copper (I) iodide (20.3 mg, 0.11 mmol) and cesium carbonate (348 mg, 1.07 mmol), in dimethylacetamide (5.3 ml), was degassed with argon for 10 minutes, heated in a sealed tube at 140 C overnight and then cooled to room temperature. Water and ethyl acetate were added, the organic layer was separated, the aqueous layer was extracted twice with ethyl acetate, the combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse CA 02813299 2013-04-17 .
phase chromatography eluting with a 1% HCl/methanol gradient provided compound 35=HCI as beige solid. MS (m/z) M+H= 519.2 Synthesis of compound 10:
OMe OMe 0 . OTBS Ph3P, DIAD, 110 0 40 TBAF I* 0 0 _____________________ 3.
OTBS OH
OH HO 1.1 OMe 28-b 11-a 28-a O, OMe I, 410 10-a + 28-b CuCCs2CO3, 1.. NH2 O
OH
N INV \N
I ' aCompound 10 Scheme 28 Step 1: Intermediate 28-a To a solution of (3-methoxyphenyl)methanol (1.38 g, 10.0 mmol) in THF
(20.0 mL) were sequentially added intermediate 11-a (2.69 g, 12.0 mmol), triphenylphosphine (3.15 g, 12.0 mmol) and DIAD (2.36 ml, 12.0 mmol) drop wise at room temperature and the reaction was then stirred overnight at room temperature. A saturated aqueous solution of ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure.
Purification by silica gel chromatography provided intermediate 28-a as a colorless oil.
Step 2: Intermediate 28-b Tetrabutylammonium fluoride trihydrate (2.88 g, 9.14 mmol) was added to a solution of intermediate 28-a (2.1 g, 6.10 mmol) in THF (10 mL) and the reaction was stirred at room temperature overnight. A saturated aqueous solution of ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 28-b as a colorless oil.
Step 3: Compound 10 A solution of intermediate 10-a (200 mg, 0.65 mmol), intermediate 28-b (225 mg, 0.97 mmol), quinolin-8-ol (16.2 mg, 0.11 mmol), copper (I) chloride (11.0 mg, 0.1 mmol) and cesium carbonate (546 mg, 1.67 mmol), in dimethylacetamide (5.5 ml), was degassed with argon for 10 minutes, heated in a sealed tube at 140 C overnight and then cooled to room temperature. Water and ethyl acetate were added, the organic layer was separated, the aqueous layer was extracted twice with ethyl acetate, the combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% HCl/methanol gradient provided compound 10.1-1C1 as a yellow solid. MS (m/z) M+H= 508.1 Synthesis of intermediate 29-i Et01=184. H2N)L= , N
, HC(0)0Et 0 toluene, reflux 0 29-a 29-c 29-b 29-c LiAIH4 Ho, j--\\_ 29-d F OMe BBr3 F OH Base F OH
TBDMSCI
=
OMe OH OTBDMS
29-e 29-f 29-g F =
TBAF
29-g Ph3P, DIAD F
HOJ , N
OTBDMS
S¨ OH
29-d 29-h 29-i Scheme 29 Step 1: Intermediate 29-b Ethyl chloroacetate, 29-a (50.0 g, 0.41 mol), and ethyl formate (30.2 g, 0.41 mol) were taken in anhydrous toluene (500 mL) and cooled to 0 C. Sodium ethoxide (35.1 g, 0.49 mol) was added portion wise. The reaction mixture was stirred at 0 C for 5 hours and then at room temperature overnight. The reaction mixture was quenched with water (250 mL) and washed twice with diethyl ether. The aqueous layer was cooled to 0 C and acidified to pH 4-5 using 1 N HCI. The aqueous layer was extracted twice with diethyl ether; the combined organic layers were dried over MgSO4 filtered and concentrated under reduced pressure to provide intermediate 29-b as beige Oil.
Step 2: Intermediate 29-c To a solution of ethyl 2-chloro-3-oxopropanoate, 29-b (34.7 g, 230 mmol), in toluene (250 ml) was added thioacetamide (26.0 g, 346.0 mmol), the reaction was stirred at 90 C overnight and then cooled to room temperature, diluted with water (300 mL) and then neutralized to pH 7 with a saturated aqueous solution of NaHCO3. Ethyl acetate was added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 29-c as beige oil.
Step 4: Intermediate 29-d To a solution of intermediate 29-c (22.2 g, 130.0 mmol) in THF (430 ml) cooled to 0 C was added a 1.0 M solution of LiAIH4in THF (91.0 ml, 91.0 mmol) and the solution was slowly warmed to room temperature and stirred for 2 hours. Water (3.5 ml) was slowly added, followed by 3.5 ml 15% NaOH
(3.5 ml) and water (10.5 ml) and the mixture was stirred for 1 hour. The reaction was filtered over celite and volatiles were removed in vacuo to provide intermediate 29-d as yellow oil.
Step 5: Intermediate 29-f To a solution of 1-fluoro-3,5-dimethoxybenzene (12.5 g, 80 mmol) in dichloromethane (80 ml), cooled to OK, was added 1.0 M solution of boron tribromide in dichloromethane (200 ml, 200 mmol), drop wise over a period of 30 minutes. The reaction was stirred for 1 hour at 0 C and then slowly warmed to room temperature and stirred for 18 hours. The reaction was cooled to 0 C and quenched by the slow addition of Me0H and water. After stirring at room temperature for 1 hour the mixture was filtered and volatiles were removed in vacuo. Ethyl acetate was added to the residue; a precipitate formed and was collected by filtration to provide intermediate 29-f as an orange solid.
Step 6: Intermediate 29-g To a solution of intermediate 29-f (10.25 g, 80.0 mmol) in DMF (50 ml), cooled to 0 C, was added imidazole (5.99 g, 88.0 mmol) and tert-butylchlorodimethylsilane (13.27 g, 88.0 mmol). The reaction was then stirred at room temperature overnight. A saturated aqueous solution of ammonium chloride and ethyl acetate were added, the organic layer was separated, washed 3 times with a saturated aqueous solution of ammonium chloride and brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 29-g as a yellow oil.
Step 7: Intermediate 29-h To a solution of intermediate 29-g (8.0 g, 33.1 mmol) and intermediate 29-d (4.70 g, 36.4 mmol) in THF (20 ml) were sequentially added triphenylphosphine (12.15 g, 46.3 mmol) and DIAD (9.0 ml, 46.3 mmol) at room temperature and the reaction was then stirred at room temperature overnight. Volatiles were removed under reduced pressure. Purification by silica gel chromatography provided intermediate 29-h as a yellow oil.
Step 8: Intermediate 29-i To a solution of intermediate 29-h (6.0 g, 16.97 mmol) in THF (85 ml) was added a 1.0 M solution of TBAF in THF (16.97 ml, 16.97 mmol) and the reaction was stirred at room temperature for 1 hour. A saturated aqueous solution of ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Diethyl ether was added to the residue; a precipitate formed and was collected by filtration to provide intermediate 29-i as white solid.
Synthesis of intermediate 30-b F = OH Ph3P/DIAD
TBAF
HON N
OTBS OTBS OH
29-g 30-a 30-b Scheme 30 Step 1: Intermediate 30-a To a solution of intermediate 29-g (9.0 g, 37.1 mmol) and 2-(methylpyrimidin-5-yl)methanol (4.61 g, 37.1 mmol) in THF (37 ml) were sequentially added triphenylphosphine (11.69 g, 44.6 mmol) and DIAD (9.39 ml, 48.3 mmol) at room temperature and the reaction was then stirred at room temperature for 4 days. Volatiles were removed under reduced pressure. Purification by silica gel chromatography provided intermediate 30-a as a yellow solid.
Step 2: Intermediate 30-b To a solution of intermediate 30-a (12.5 g, 35.9 mmol) in THF (72 ml) was added a 1.0 M solution of TBAF in THF (35.9 ml, 35.9 mmol) and the reaction was stirred at room temperature for 1 hour. A saturated aqueous solution of ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 30-b as a white solid.
Synthesis of intermediate 31-d LiAIH4 31-a 31-b F = OH Ph3P, DD F =01 TBAF
F tio OTBS % OTBS OH
29-g 31-c 31-d Scheme 31 Step 1: Intermediate 31-b To a solution of methyl 6-methylnicotinate 31-a (20.10 g, 133 mmol) in THF
(90 ml) cooled to 0 C was added drop wise a 1.0 M solution of LiAIH4 in THF
(100 ml, 100 mmol) and the reaction was then stirred at 0 C for 1 hour.
Water (3.8 ml) was slowly added, followed by 15% NaOH (3.5 ml) and water (11.4 ml) and the mixture was stirred at room temperature for 1 hour. The reaction was filtered over celite and volatiles were removed in vacuo to provide intermediate 31-b as a yellow oil.
Step 2: Intermediate 31-c To a solution of intermediate 29-g (13.2 g, 54.5 mmol) and intermediate 31-b (7.38 g, 59.9 mmol) in THF (50 ml) were sequentially added triphenylphosphine (21.43 g, 82.0 mmol) and DIAD (17.10 ml, 87.0 mmol) at room temperature and the reaction was then stirred at room temperature for 1 hour. Volatiles were removed under reduced pressure. Purification by silica gel chromatography provided intermediate 31-c as a colorless oil.
Step 3: Intermediate 31-d To a solution of intermediate 31-c (7.6 g, 21.87 mmol) in THF (44 ml) was added tetrabutylammonium fluoride trihydrate (5.72 g, 21.87 mmol) and the reaction was stirred at room temperature for 1 hour. Saturated aqueous ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 31-d as white solid.
Synthesis of intermediate 32-f O rN\\ NaBH4, CaC,I2 OrN1\\ /OH
\--/
32-a 32-b ¨0 PPTS
0, / 0, ¨N\\ _) \-1 OH
HO
32-b 32-c 32-d F io OH Ph3P, DIAD TBAF
TBSO 40. ____________ - HO 40 OTBS 32-d 0 0 N /
29-g 32-e 0 o 32-f 0 Scheme 32 Step 1: Intermediate 32-b To a solution of dimethyl pyridine-2,5-dicarboxylate (13.0 g, 66.6 mmol) in a mixture of THF (110 mL) and ethanol (110 mL) was added calcium chloride (29.6 g, 266 mmol). After stirring at room temperature for 30 minutes the reaction was cooled to 0 C and sodium borohydride (3.78 g, 100 mmol) was added portion wise. After the addition was completed the reaction was stirred at room temperature overnight. A saturated aqueous solution of ammonium chloride and dichloromethane were added, the organic layer was separated and the aqueous phase was extracted twice with dichloromethane. The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure to provide intermediate 32-b as a yellow solid.
Step 2: Intermediate 32-c To a solution of intermediate 32-b (1.70 g, 10.17 mmol) in dichloromethane (203 mL) was added 3,4-dihydro-2H-pyran (4.28 g, 50.8 mmol) and PPTS
(2.56 g, 10.17 mmol) and the reaction was stirred at room temperature overnight. Water was added and the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure to provide intermediate 32-c as a white solid.
Step 3: Intermediate 32-d To a solution of intermediate 32-c (2.56 g, 10.17 mmol) in THF (51 ml) cooled to 0 C was added drop wise a 1.0 M solution of DIBALH in hexane (23.39 ml, 23.39 mmol) and the reaction was then stirred at 0 C for 1.5 hour and room temperature overnight. Water (1.0 ml) was slowly added, followed 15% NaOH (3.5 ml) and water (2.3 ml) and the mixture was stirred at room temperature for 30 minutes. The reaction was filtered over celite and volatiles were removed under reduced pressure. Purification by silica gel chromatography provided intermediate 32-d as a yellow oil.
Step 4: Intermediate 32-e To a solution of intermediate 29-g (1.57 g, 6.51 mmol) and intermediate 32-d (2.56 g, 7.17 mmol) in THF (7 ml) were sequentially added triphenylphosphine (2.56 g, 9.77 mmol) and DIAD (2.04 ml, 10.42 mmol) at room temperature and the reaction was then stirred at room temperature overnight. Volatiles were removed under reduced pressure. Purification by silica gel chromatography provided intermediate 32-e as a yellow solid.
Step 5: Intermediate 32-f To a solution of intermediate 32-e (2.2 g, 4.91 mmol) in THF (9.8 ml) was added a 1.0 M solution of TBAF in THF (4.91 ml, 4.91 mmol) and the reaction was stirred at room temperature for 1 hour. A saturated aqueous solution of ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 32-f as a white solid.
Synthesis of intermediate 33-a -o riY1 y W
=MCP BA _ F 1Wa& 0,....)-OTBS OH
31-c 33-a Scheme 33 To a solution of intermediate 31-c (424 mg, 1.82 mmol) in dichloromethane (9.0 ml) was added m-CPBA (538 mg, 2.18 mmol) and the reaction was stirred at room temperature for 4 hours. A saturated aqueous solution of NaHCO3 and dichloromethane were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 33-a as a white solid.
Synthesis of intermediate 34-d N N
tBuONa ,,E
.,.,1 F 0 F K2c03 F 0 F id o toluene/DMPU F=
0 0 N HC!
F ir N
1 ___________________________________________________________________ \
MOMCIN
OH OMOM HO r¨, \.... OMOM OH
N
34-a 34-b I 34-c 34-d Scheme 34 Step 1: Intermediate 34-b To a solution of 3,5-difluorophenol (15.0 g, 115 mmol) in acetone (200 ml) was added K2CO3 (23.90 g, 173 mmol) and bromomethyl methyl ether (15.85 g, 127 mmol). The reaction was then stirred at room temperature overnight and filtered. The filtrate was concentrated under reduced pressure to provide intermediate 34-b as a colorless oil. , Step 2: Intermediate 34-c To a solution of (1-methyl-1H-imidazol-5-y1) methanol (3.1 g, 27.6 mmol) and intermediate 34-b (4.01 g, 23.04 mmol) in toluene (25.0 ml) and DMPU
(25.0 ml) was added sodium 2-methylpropan-2-olate (4.43 g, 46.1 mmol).
The reaction was stirred overnight at 80 C and then cooled to room temperature. A saturated aqueous solution of ammonium chloride and ethyl acetate were added, the organic layer was separated, washed twice with a saturated aqueous solution of ammonium chloride and brine, dried over MgSO4, filtered and concentrated in vacuo. Purification by silica gel chromatography provided intermediate 34-c as beige oil.
Step 3: Intermediate 34-d To a solution of intermediate 34-c (3.2 g, 12.02 mmol) in Me0H (25.0 ml) was added 4N HCI in dioxane (10.95 ml, 361.0 mmol) and the reaction was stirred overnight at room temperature. Volatiles were removed in vacuo.
Diethyl ether was added to the residue; a precipitate formed and was collected by filtration to provide intermediate 34-d-FICI as a white solid.
Synthesis of intermediate 35-d LiAIH4 HOJS
35-a 35-b tBua F F
toluONene/DMPU
HCI
, N
OMOM OMOM OH
34-h 35-c 35-d Scheme 35 Step I: Intermediate 35-b To a solution of 1,2-dimethy1-1H-imidazole-5-carbaldehyde (1.0 g, 8.06 mmol) in THF (40.3 mL) cooled to 0 C was added drop wise a 1.0 M solution of LiAlHain THF (6.04 ml, 6.04 mmol) and the reaction was then stirred at room temperature for 1 hour. Water (250 uL) was slowly added, followed by 15% NaOH (250 uL) and water (750 uL) and the mixture was stirred at room temperature for 1 hour. The reaction was filtered over celite and volatiles were removed in vacuo to provide intermediate 35-b as a white solid.
Step 2: Intermediate 35-c To a solution of intermediate 35-b (1.50 g, 11..89 mmol) and intermediate 34-b (2.07 g, 11.89 mmol) in DMPU (11.89 mL) and toluene (11.89 mL) was added sodium 2-methylpropan-2-olate (3.43 g, 35.7 mmol) at room temperature. The reaction was stirred overnight at 80 C and then cooled to room temperature. A saturated aqueous solution of ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure.
Purification by silica gel chromatography provided intermediate 35-c as a yellow oil.
Step 3: Intermediate 35-d To a solution of intermediate 35-c (3.30 g, 11.77 mmol) in Me0H (36.2 mL) was added 4N HCI in dioxane (10.7 mL, 353 mmol) and the reaction was stirred at room temperature overnight. Volatiles were removed under reduced pressure. Diethyl ether was added to the residue; a precipitate formed and was collected by filtration to provide intermediate 35-d=HCI as a white solid.
Synthesis of intermediate 36-f N ,N
n ,'-O--H
__________________________ .athj ,-SH
OH KSCN HON H2W04, H20.
HO,..f -N
0H0 Acetic acid ?
?
36-a 112N---- WI
36-b = 114 36-c 36-d N N
tBuONa o j-toluene/DMPU
__________________________________________________ ..
IW-36-d 34-b 36-e 0 36-f It Scheme 36 Step 1: Intermediate 36-c To a suspension of 2-(benzyloxy)ethanamine HCI, 36-b (2.08 g, 11.10 mmol), and 2,5-bis(hydroxymethyl)-1,4-dioxane-2,5-diol 36-a (2.00 g, 11.10 mmol) in iPrOH (8 mL), were sequentially added potassium thiocyanate (1.62 g, 16.7 mmol) and acetic acid (2.03 mL, 35.5 mmol) drop wise. The mixture was stirred at room temperature overnight. Water was added; a precipitate formed and was collected by filtration to provide intermediate 36-c as a white solid.
Step 2: Intermediate 36-d To a solution of intermediate 36-c (1.5 g, 5.67 mmol) and H2W04 (14 mg, 0.057 mmol) in Me0H (22.7 mL) at 40 C was added H202 (1.85 mL, 18.16 mmol) drop wise. The mixture was stirred at reflux overnight and then cooled to room temperature. Volatiles were removed under reduced pressure. Purification by silica gel chromatography provided intermediate 36-d as a colorless oil.
Step 3: Intermediate 36-e To a solution of intermediate 36-d (1.46 g, 6.32 mmol) and intermediate 34-b (1.0 g, 5.74 mmol) in DMPU (11.48 ml) and toluene (11.48 ml) was added sodium 2-methylpropan-2-olate (1.10 g, 11.48 mmol) at room temperature.
The reaction was stirred overnight at 80 C and then cooled to room temperature. A saturated aqueous solution of ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure.
Purification by silica gel chromatography provided intermediate 36-e as a colorless oil.
Step 4: Intermediate 36-f To a solution of intermediate 36-e (400 mg, 1.03 mmol) in Me0H (10.4 mL) was added 4N HCI in dioxane (2.50 mL, 10.0 mmol) the reaction was stirred at room temperature overnight. Volatiles were removed under reduced pressure to provide intermediate 364.1-1C1 as a white solid.
Synthesis of intermediates 37-f and 37-f' SiMe3 SiMe3 N r r_N r NaBH4 NaH (:).M + N HOõ.N, +
N
Me3S1 ' Oji 0) HO,(Ntl H (3'CI 0 ,) 37-a 37-h 37-b 37-c 37-d SiMe3 SiMe3 TMS
( tBuONa F toluene/DMPU --N N,---- \
F 0 F K2CO3 F io cc:
.
F F =
0-- \
OH Br 0 OBn 37-c + 37-d w- to L-TMS
34-a 37-d 0 37-e 0 37-e' TMS
( (N____\\ N,:-- \
r 37-e + 37e' H2, Pd/C N
----.- +
F i, o 1.1 ---\---TMS
HO 374 HO 37-f Scheme 37 Step 1: Intermediates 37-b and 37-b' To a solution of 1H-imidazole-5-carbaldehyde, 37-a (3.0 g, 31..2 mmol), in DMF (20 ml) was added a 60% dispersion of NaH in mineral oil (1.25 g, 31.2 mmol) portion wise. After stirring for 30 minutes at room temperature, (2-(chloromethoxy)ethyl)trimethylsilane (5.73 g, 34.3 mmol) was added and the reaction was then was stirred at room temperature overnight. A
saturated aqueous solution of ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediates 37-b and 37-b' as an inseparable mixture.
Step 2: Intermediates 37-c and 37-c' To a solution of intermediates 37-b and 37-b' (3.2 g, 14.14 mmol) in THF
(56.6 ml) was added NaBH4 (535 mg, 14.14 mmol) at room temperature.
The reaction was stirred overnight at room temperature and then cooled to 0 C. A saturated aqueous solution of ammonium chloride and ethyl acetate were added, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate. The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure to provide intermediate 37-c and 37-c' as an inseparable mixture.
Step 3: Intermediate 37-d To a solution of 3,5-difluorophenol (8.0 g, 61.4 mmol) in acetone (100 ml) was added potassium carbonate (17.0 g, 123.0 mmol) and potassium iodide (1.021 g, 6.15 mmol). The reaction was heated to 65 C and benzyl bromide (8.03 g, 67.6 mmol) was added. The reaction was then stirred overnight at 65 C, cooled to room temperature and filtered. Volatiles were removed in vacuo. A saturated aqueous solution of ammonium chloride and ethyl acetate were added to the residue, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated in vacuo. Purification by silica gel chromatography provided intermediate 37-d as a colorless oil.
Step 4: Intermediate 37-e and 37-e' To a solution of intermediates 37-c and 37-c' (1.0 g, 4.38 mmol) and intermediate 37-d (877 mg, 3.98 mmol) in DMPU (7.96 ml) and toluene (7.96 ml) was added sodium 2-methylpropan-2-olate (765 mg, 3.98 mmol) at room temperature. The reaction was stirred overnight at 80 C and then cooled to room temperature. A saturated aqueous solution of ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediates 37-e and 37-e' as an inseparable mixture.
Step 5: Intermediates 37-f and 37-f' A methanol solution of intermediate 37-e and 37-e' (140 mg, 0.32 mmol) was treated with 10% palladium on carbon (70 mg, 0.045 mmol) and purged with H2. The solution was stirred under H2 (1 atm) for 2 hours before being filtered through celite. The filtrate was concentrated in vacuo to provide intermediate 37-f and 37-f' as an inseparable mixture.
Synthesis of intermediates 38-e and 38-e' Me3Si Me3Si N
i O HO,Z¨N¨
N
isi'---N NaBH4 NaH H o) + )+ <o ---- ___________________________________________ a OyfN ' H
H 38-h Me3Si,...,,O.,.C1 N--( N--( 4 38-h 38-c 38-d 'l SiMe3 SiMe3 y 38-a "H NO
TMS
(o tBuOK ( N--7----F 0 F toluene/DMPU N--(' F
r)N 0 _____________________ . +
F
0 0 --\-_ 38-c + 38-c' TMS
tw- 0 37-d 00:1 0 38-d 0 38-d' 40 I.
TMS
(o (N4 N=---H2, Pd/C rk,N r ,./../N---\0 38-d + 38-d _______ . +
F00 F 40 0 --\__.
TMS
OH 38-e HO 38-e' Scheme 38 Step 1: Intermediates 38-b and 38-b' To a solution of 1H-imidazole-5-carbaldehyde (5.0 g, 45.4 mmol) in DMF (20 mL) was added a 60% dispersion of NaH mineral oil (1.81 g, 45.4 mmol) portion wise. After stirring for 30 minutes at room temperature, (2-(chloromethoxy)ethyl)trimethylsilane (9.08 g, 54.5 mmol) was added and the reaction was then stirred at room temperature overnight. A saturated aqueous solution of ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediates 38-b and 38-h' as an inseparable mixture.
Step 2: Intermediates 38-c and 38-c' To a solution of intermediate 38-b and 38-b' (7.0 g, 29.1 mmol) in THF
(116.0 ml) was added NaBH4 (1.10 g, 29.1 mmol) at room temperature. The reaction was stirred overnight at room temperature and then cooled to 0 C.
A saturated aqueous solution of ammonium chloride and ethyl acetate were added, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate. The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure to provide intermediates 38-c and 38-c' as an inseparable mixture.
Step 3: Intermediate 38-d and 38-d' To a solution of intermediate 38-c and 38-c' (1.0 g, 4.13 mmol) and intermediate 37-d (826 mg, 3.75 mmol) in DMPU (7.50 ml) and toluene (7.50 ml) was added sodium 2-methylpropan-2-olate (721 mg, 7.50 mmol) at room temperature. The reaction was stirred overnight at 80 C and then cooled to room temperature. A saturated aqueous solution of ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediates 38-d and 38-d' as an inseparable mixture.
Step 4: Intermediates 38-e and 38-e' A methanol solution of intermediates 38-d and 38-d' (200 mg, 0.45 mmol) was treated with 10% palladium on carbon (96 mg, 0.045 mmol) and purged with H2. The solution was stirred under H2 (1 atm) for 2 hours before being filtered through celite. The filtrate was concentrated in vacuo to provide intermediate 38-e and 38-e' as an inseparable mixture.
Synthesis of intermediate 39-b F = OH Ph3P, DD F = F
TBAF
HON
OTBS \ c \ OTBS OH
29-g 39-a 39-b Scheme 39 Step 1: Intermediate 39-a To a solution of intermediate 29-g (4.20 g, 17.3 mmol) and pyrimidin-5-ylmethanol (1.90 g, 17.3 mmol) in THF (35 mL) were sequentially added triphenylphosphine (5.91 g, 22.5 mmol) and DIAD (4.38 mL, 22.5 mmol) at room temperature and the reaction was then stirred at room temperature for 3 hours. Volatiles were removed under reduced pressure. Purification by silica gel chromatography provided intermediate 39-a as a white solid.
Step 2: Intermediate 39-b To a solution of intermediate 39-a (5.80 g, 17.3 mmol) in THF (35 mL) was added a 1.0 M solution of TBAF in THF (17.3 ml, 17.3 mmol) and the reaction was stirred at room temperature for 1 hour. A saturated aqueous solution of ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 39-b as a white solid.
Synthesis of intermediate 40-b rN,n F OH Ph3P, DIAD F = 0,,N) TBAF F =
ON) HO
OTBS \ OTBS OH
29-g 40-a 40-b Scheme 40 Step 1: Intermediate 40-a To a solution of intermediate 29-g (4.62 g, 19.1 mmol) and pyrazin-2-ylmethanol (2.10 g, 19.1 mmol) in THF (38 mL) were sequentially added triphenylphosphine (7.50 g, 28.6 mmol) and DIAD (5.19 ml, 26.7 mmol) at room temperature and the reaction was then stirred at room temperature overnight. Volatiles were removed under reduced pressure. Purification by silica gel chromatography provided intermediate 40-a as a colorless oil.
Step 2: Intermediate 40-b To a solution of intermediate 40-a (3.40 g, 10.2 mmol) in THF (20 mL) was added a 1.0 M solution of TBAF in THF (10.2 ml, 10.2 mmol) and the reaction was stirred at room temperature for 1 hour. A saturated aqueous solution of ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 40-b as a white solid.
Synthesis of intermediate 41-b F = OH Ph3P, DIAD F io 0 lel TBAF F = 0 I.
HO
OTBS OTBS OH
29-g 41-a 41-b Scheme 41 Step 1: Intermediate 41-a To a solution of intermediate 29-g (2.60 g, 10.7 mmol) and benzyl alcohol (1.39 g, 12.9 mmol) in THF (20 mL) were sequentially added triphenylphosphine (3.94 g, 15.02 mmol) and DIAD (2.92 mL, 15.0 mmol) at room temperature and the reaction was then stirred at room temperature overnight. Volatiles were removed under reduced pressure. Purification by silica gel chromatography provided intermediate 41-a as colorless oil.
Step 2: Intermediate 41-b To a solution of intermediate 41-a (1.40 g, 4.21 mmol) in THF (10 ml) was added a 1.0 M solution of TBAF in THF (4.63 ml, 4.63 mmol) and the reaction was stirred at room temperature overnight. A saturated aqueous solution of ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 41-b as a colorless oil.
Synthesis of intermediate 42-d Boc NNH NaBH3CN HN,NH2 0=0 ______________________ HCI HCI
,N
H2N , Boc 42-a 42-b 42-c Br Br TEA
NC NC
N
CN 42-c 9-b 42-d Scheme 42 Step 1: Intermediate 42-b To a solution of cyclopentanone (10.00 g, 119.0 mmol) in Me0H (594 mL), were added tert-butyl hydrazinecarboxylate (16.50 g, 125.0 mmol) and the reaction was stirred overnight at room temperature. Volatiles were removed under reduced pressure to provide intermediate 42-b as a white solid.
Step 2: Intermediate 42-c To a solution of intermediate 42-b (10.00 g, 50.40 mmol) in THF (50.4 mL) and Me0H (50.4 mL) was added sodium cyanoborohydride (3.80 g, 60.5 mmol) portion wise. The reaction was refluxed under argon for 10 minutes, and then cooled to room temperature. 6N NCI (25 mL) was added, the mixture was refluxed for 3 hours, cooled to room temperature and stirred overnight. The reaction was filtered to remove inorganic insoluble material and the filtrate was concentrated under reduced pressure and azeotroped three times with toluene. The residue was dissolved in hot isopropanol, cooled to room temperature, diluted with ether and then cooled to 0 C. A
precipitate formed and was collected by filtration to provide intermediate 42-c=FICI as a white solid.
Step 3: Intermediate 42-d To a solution of intermediate 9-b (3.00 g, 11.4 mmol) and TEA (3.50 mL, 25.1 mmol) in Et0H (11.4 mL) was added intermediate 42-c=HCI (1.86 g, 13.7 mmol) and the reaction was then stirred for 2 hours at 100 C. Volatiles were removed under reduced pressure. A saturated aqueous solution of ammonium chloride and ethyl acetate were added to the residue, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 42-d as a white solid.
Synthesis of intermediate 43-e Boc Boc NI' NH
HN
H Pd/CNH HCI HNNH2 0\ 2 . _______________ . HCI
N, H2N Boc 43-a 43-b 43-c 43-d Br Br TEA
NC NC
N
CN 43-d 9-b 43-e Scheme 43 Step 1: Intermediate 43-b To a solution of dihydro-2H-pyran-4-(3H)-one (15.0 g, 150.0 mmol) in Me0H
(749 mL), were added tert-butyl hydrazinecarboxylate (20.79 g, 157.0 mmol) and the reaction was stirred overnight at room temperature. Volatiles were removed under reduced pressure to provide intermediate 43-b as a white solid.
Step 2: Intermediate 43-c A methanol solution of intermediate 43-b (32.1 g, 150.0 mmol) was treated with 10% palladium on carbon (6.39 g, 3.00 mmol), acetic acid (100 pL) and purged with H2. The solution was stirred under H2 (1 atm) overnight before being filtered through celite. The filtrate was concentrated in vacuo to provide intermediate 43-c as a white solid.
Step 3: Intermediate 43-d To a solution of intermediate 43-c (32.4 g, 150 mmol) in Me0H (300 mL) was added 4N HCI in 1,4-dioxane (300 ml, 1200 mmol) and the reaction was stirred at room temperature for 5 hours. Diethyl ether was added and a precipitate formed which was collected by filtration to provide intermediate 43-d=HCI as a white solid.
Step 4: Intermediate 43-e To a solution of intermediate 9-b (5.00 g, 19.0 mmol) and TEA (5.30 mL, 38.0 mmol) in Et0H (19.0 mL) was added intermediate 43-c=FICI (3.48 g, 22.81 mmol) and the reaction was then stirred for 2 hours at 100 C.
Volatiles were removed under reduced pressure. A saturated aqueous solution of ammonium chloride and ethyl acetate were added to the residue, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure to provide intermediate 43-e as a yellow solid.
Synthesis of intermediate 44-d Boc NNH
H _________________________ A. I
V\ NaBH3CN
HCI HN,NH2 _________________________________________________ p riN
N, H2N, Boc 44-a 44-b 44-c Br Br SI ilk TEA
NC ________________________________________ A NC
0 I \ N
44-c CN
H2N "....,,_ 9-b 44-d Scheme 44 Step 1: Intermediate 44-b tert-Butyl hydrazinecarboxylate (7.60 g, 57.5 mmol) was added to acetone (50 mL) and the reaction was stirred overnight at room temperature.
Volatiles were removed under reduced pressure to provide intermediate 44-b as a white solid.
Step 2: Intermediate 44-c To a solution of intermediate 44-b (9.90 g, 57.5 mmol) in THF (57.5 mL) and Me0H (57.5 mL) was added sodium cyanoborohydride (4.34 g, 69.0 mmol) portion wise. The reaction was refluxed under nitrogen for 10 minutes, and then cooled to room temperature. 6N HCI (30 mL) was added, the mixture was refluxed for 3 hours, cooled to room temperature and stirred overnight.
The reaction was filtered to remove inorganic insoluble material and the filtrate was concentrated under reduced pressure and azeotroped three times with toluene for complete water removal. The residue was dissolved in hot isopropanol, cooled to room temperature, diluted with ether and then cooled to 0 C. A precipitate formed and was collected by filtration to provide intermediate 44-c=HCI as a white solid.
Step 3: Intermediate 44-d To a solution of intermediate 9-b (12.61 g, 47.9 mmol) and TEA (14.70 mL, 105.0 mmol) in Et0H (96.0 ml) was added intermediate 44-c.HCI (6.36 g, 57.5 mmol) and the reaction was then stirred for 2 hours at 100 C. Volatiles were removed under reduced pressure. A saturated aqueous solution of ammonium chloride and ethyl acetate were added to the residue, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 44-d as a white solid.
Synthesis of intermediate 45-a Br Br TEA
NC __________________________________________________ . NC410 / 0 \
I N
CN) NHNH2 N
/)----9-b 45-a Scheme 45 To a solution of intermediate 9-b (2.0 g, 7.60 mmol) and TEA (2.12 ml, 15.2 mmol) in Et0H (7.60 mL) was added tert-butylhydrazine hydrochloride (1.13 g, 9.12 mmol) and the reaction was then stirred for 2 hours at 100 C.
Volatiles were removed under reduced pressure. A saturated aqueous solution of ammonium chloride and ethyl acetate were added to the residue, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure to provide intermediate 45-a as a yellow solid.
Synthesis of intermediate 46-a Br Br TEA
NC NC
0 \ N
9-b 46-a Scheme 46 To a solution of intermediate 9-b (1.45 g, 5.53 mmol) and TEA (1.54 mL, 11.1 mmol) in Et0H (15.0 mL) was added cyclohexylhydrazine hydrochloride (1.00 g, 6.64 mmol) and the reaction was then stirred for 2 hours at 100 C.
Volatiles were removed under reduced pressure. A saturated aqueous solution of ammonium chloride and ethyl acetate were added to the residue, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure to provide intermediate 46-a as a yellow solid.
Synthesis of intermediate 47-a Br Br TEA
NC NC
0 I \ N
CN HO H2 H2N Nv_ 9-b 47-a OH
Scheme 47 To a solution of intermediate 9-b (2.00 g, 7.60 mmol) and TEA (1.27 mL, 9.12 mmol) in Et0H (7.60 mL) was added 2-hydroxyethylhydrazine (618 pL, 9.12 mmol) and the reaction was then stirred for 2 hours at 100 C. Volatiles were removed under reduced pressure. A saturated aqueous solution of ammonium chloride and ethyl acetate were added to the residue, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure to provide intermediate 47-a as a white solid.
Synthesis of intermediate 48-c cK)OH NaOH
Hydrazine 48-a 48-b Br Br TEA
NC NC
N
48-b 9-b 48-c OH
Scheme 48 Step 1: Intermediate 48-b To a mixture of sodium hydroxide (7.37 g, 184.0 mmol) and hydrazine monohydrate (46.10 g, 921.0 mmol) heated to 95 C, was added 1-chloro-2-methylpropan-2-ol (20.00 g, 184.0 mmol). The reaction was stirred overnight at 95 C and then cooled to room temperature. Volatiles were removed under reduced pressure. THF (40 mL) and diethyl ether (40 mL) were added to the residue; a precipitate formed which was removed by filtration. The filtrate was concentrated under reduced pressure to provide intermediate 48-b as a colorless oil.
Step 2: Intermediate 48-c To a solution of intermediate 9-b (4.45 g, 16.9 mmol) and TEA (4.71 mL, 33.8 mmol) in Et0H (15.0 mL) was added intermediate 48-b (1.76 g, 16.9 mmol) and the reaction was then stirred for 2 hours at 100 C. Volatiles were removed under reduced pressure. A saturated aqueous solution of ammonium chloride and ethyl acetate were added to the residue, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure to provide intermediate 48-c as a white solid.
Synthesis of intermediate 49-c NaOH
NOH
Hydrazine 49-a 49-b Br TEA Br NC N
N
49-h CN N
9-b 49-c OH
Scheme 49 Step 1: Intermediate 49-b To a mixture of sodium hydroxide (1.15 g, 28.8 mmol) and hydrazine monohydrate (7.20 g, 144.0 mmol) heated to 95 C, was added 3-bromo-1-propanol (4.00 g, 28.8 mmol) and the reaction was stirred overnight at 95 C
and then cooled to room temperature. Volatiles were removed under reduced pressure. Ethanol was added to the residue; a precipitate formed and was removed by filtration. The filtrate was concentrated under reduced pressure and 1M HCI in diethyl was added to the residue. After stirring for 15 minutes a precipitate formed and was collected by filtration to provide intermediate 49-b=HCI as a white solid.
Step 2: Intermediate 49-c To a solution of intermediate 9-b (1.12 g, 4.28 mmol) and TEA (1.19 mL, 8.56 mmol) in Et0H (10.0 mL) was added intermediate 49-b=FICI (650 mg, 5.13 mmol) and the reaction was then stirred for 2 hours at 100 C. Volatiles were removed under reduced pressure. A saturated aqueous solution of ammonium chloride and ethyl acetate were added to the residue, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure to provide intermediate 49-c as a white solid.
Synthesis of intermediate 50-c Boc n OH FIN,N
NaBH3CN H2N,INH HCI
_____________________________ _ ____________________ .
H
HCI
r-OH N
H2NõBoc OH OH OH OH
36-a 50-a 50-b Br Br TEA
___________________________ .
CN 50-b N
9-b OH 50-c Scheme 50 Step 1: Intermediate 50-a Dihydroxyacetone dimer (15.0 g) and tert-butyl hydrazinecarboxylate (22.01 g) were dissolved in ethanol (500 mL) and this solution was stirred at room temperature for 2 days. After the reaction mixture was concentrated under reduced pressure, the resulting residue was recrystallized from ethyl acetate to provide intermediate 50-a as a white solid.
Step 2: Intermediate 50-b To a solution of intermediate 50-a (10.0 g, 49.0 mmol) in THF (49.0 mL) and Me0H (49.0 mL) was added sodium cyanoborohydride (3.69 g, 58.8 mmol) portion wise. The reaction was refluxed under nitrogen for 10 minutes, and then cooled to room temperature. 6N HCI (40 mL) was added, the mixture was refluxed for 3 hours, cooled to room temperature and stirred overnight.
The reaction was filtered to remove inorganic insoluble material and the filtrate was concentrated under reduced pressure and azeotroped three times with toluene to provide intermediate 50-b=HCI as a white solid.
Step 3: Intermediate 50-c To a solution of intermediate 9-b (10.70 g, 40.9 mmol) and TEA (12.5 mL, 90.0 mmol) in Et0H (40.9 mL) was added intermediate 50-13.1-1C1 (7.00 g, 49.1 mmol) and the reaction was then stirred for 2 hours at 100 C. Volatiles were removed under reduced pressure. A saturated aqueous solution of ammonium chloride and ethyl acetate were added to the residue, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 50-c as a beige solid.
Synthesis of intermediate 51-a Br Br O fa Ph3P, DIAD
NC ________________________________ v NC
l \ N l \ N
H2N [\il HC:Y>C0 H2N
LC) 9-c 51-a Scheme 51 To a solution of intermediate 9-c (1.40 g, 5.32 mmol) and 3-methyloxetan-3-yl)methanol (1.08 g, 10.64 mmol) in THF (5.3 mL) were sequentially added triphenylphosphine (1.67 g, 6.39 mmol) and DIAD (1.13 mL, 5.85 mmol) at room temperature and the reaction was then stirred at room temperature for 4 days. Volatiles were removed under reduced pressure. Purification by silica gel chromatography provided intermediate 51-a as a white solid.
Synthesis of intermediate 52-a NC Br Br Ph3P, DIAD NC
H2N /\N H
1\1 rN N,N
0) 9-c 52-a 170 Scheme 52 To a solution of intermediate 9-c (500 mg, 1.90 mmol) and 3-morpholinopropan-1-ol (263 pl, 1.90 mmol) in THF (19.0 ml) cooled to 0 C
were sequentially added triphenylphosphine (498 mg, 1.90 mmol) and DIAD
(370 pl, 1.90 mmol). The reaction was stirred at 0 C for 1 hour and room temperature overnight. Volatiles were removed under reduced pressure.
Purification by silica gel chromatography provided intermediate 52-a as a white foam.
Synthesis of intermediate 53-a Br Br NH2 = NH2 N Ph3P, DIAD
,N N
N N N iN\
r-N\ 10 9-d 53-a Scheme 53 To a solution of intermediate 9-d (3.00 g, 10.3 mmol) and N-(2-hydroxyethyl)morpholine (1.88 ml, 15.1 mmol) in THF (103 ml) cooled to 0 C were sequentially added triphenylphosphine (4.07 mg, 15.1 mmol) and DIAD (3.02 ml, 15.5 mmol). The reaction was stirred at 50 C overnight.
Volatiles were removed under reduced pressure. Purification by silica gel chromatography provided intermediate 53-a as a white solid.
Synthesis of intermediate 54-a Br Br = 110+
Ph3P, DIAD NC
NC
\ /
9-c 54-a Scheme 54 To a solution of intermediate 9-c (500 mg, 1.90 mmol) and 2-(pyrrolidin-1-yl)ethanol (219 mg, 1.90 mmol) in THF (9.5 ml) cooled to 0 C were sequentially added triphenylphosphine (498 mg, 1.90 mmol) and DIAD (370 pl, 1.90 mmol). The reaction was stirred at 0 C for 1 hour and room temperature for 30 minutes. Volatiles were removed under reduced pressure.
Purification by silica gel chromatography provided intermediate 54-a as yellow solid.
Synthesis of Compound 65:
Br 0 0 it H2N N)-(OH 0 Cul, Cs2CO3 NH2 I' N 31-d N N
bo 17-a Compound 65 Scheme 55 To a solution of intermediate 17-a (321 mg, 0.85 mmol) and intermediate 31-d (200 mg, 0.85 mmol) in 1,4-dioxane (4.30 ml) were sequentially added N,N-dimethylglycine (265 mg, 2.57 mmol), copper(I) iodide (163 mg, 0.85 mmol) and cesium carbonate (1.12 g, 3.43 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate. The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 65.2HCI as a white solid.
Synthesis of Compound 85 Br 0 0 411t NC * Cul, Cs2CO3 tN
HC(OMe)3 =
/ \ N
_______________________________________________ =
/
H2N m 31-d NC N
N
' )\ H2N N
44-d 56-a Compound Scheme 56 Step 1: Intermediate 56-a To a solution of intermediate 44-d (5.00 g, 16.4 mmol) and intermediate 31-d (4.20 g, 18.0 mmol) in 1,4-dioxane (54.6 ml) were sequentially added N,N-dimethylglycine (3.80 g, 36.9 mmol), copper(I) iodide (2.34 g, 12.29 mmol) and cesium carbonate (21.35 g, 65.5 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate.
The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided intermediate 56-a.HCI as a white solid.
Step 2: Compound 85 Intermediate 56-a=FICI (3.13 g, 6.84 mmol) and trimethyl orthoformate (48.7 ml, 445.0 mmol) were heated at 110 C for 3 hours. Excess trimethyl orthoformate was removed in vacuo and the residue was treated with ammonia (7.0 N in Me0H) (48.9 ml, 342.0 mmol). The mixture was stirred at room temperature for 3 days and volatiles were removed under reduced pressure. Purification by reverse phase chromatography eluting with a 1%
aqueous HCl/methanol gradient provided compound 85.2HCI as a white solid.
MS (m/z) M+H= 485.2 Synthesis of Compound 91:
Br Oil 0 0 0 =
NC Cul, Cs2CO3 HC(OMe)3 40 _______________________________________________ =
H \ N \
¨2..N N- 33-a NC N NH3 N
\
) 43-e 57-a 0 Compound 91 Scheme 57 Step 1: Intermediate 57-a To a solution of intermediate 43-e (192 mg, 0.55 mmol) and intermediate 33-a (190 mg, 0.66 mmol) in 1,4-dioxane (2.8 ml) were sequentially added N,N-dimethylglycine (129 mg, 1.24 mmol), copper(I) iodide (79 mg, 0.42 mmol) and cesium carbonate (722 mg, 2.21 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate.
The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided intermediate 57-a as a yellow foam.
Step 2: Compound 91 Intermediate 57-a (286 mg, 0.55 mmol) and trimethyl orthoformate (3.94 ml, 36.0 mmol) were heated at 110 C for 1 hour. Excess trimethyl orthoformate was removed in vacuo and the residue was treated with 7.0 N
ammonia in Me0H (3.96 ml, 27.7 mmol). The mixture was stirred at room temperature for 3 days and volatiles were removed under reduced pressure.
Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 91.HCI as white solid. MS (m/z) M+H= 543.1 Synthesis of Compound 101:
Br Cul, Cs2CO3HC(OMe)3 /\ 0 1. 0 NH2' H2N NN NH3 "N
32-f \
N NiN OH
NC NN
43-eH2N 58-a o Compound 101 o Scheme 58 Step 1: Intermediate 58-a To a solution of intermediate 43-e (400 mg, 1.15 mmol) and intermediate 32-f (384 mg, 1.15 mmol) in 1,4-dioxane (2.8 ml) were sequentially added N,N-dimethylglycine (267 mg, 2.59 mmol), copper(I) iodide (165 mg, 0.86 mmol) and cesium carbonate (1.50 g, 4.61 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate. The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure to provide intermediate 58-a as a beige foam.
Step 2: Compound 101 Intermediate 58-a (690 mg, 1.15 mmol) and trimethyl orthoformate (8.18 ml, 74.8 mmol) were heated at 110 C for 4 days. Excess trimethyl orthoformate was removed in vacuo and the residue was treated with 7.0 N
ammonia in Me0H (8.21 ml, 57.5 mmol). The mixture was stirred at room temperature overnight and volatiles were removed under reduced pressure.
Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 101.2HCI as a white solid. MS
(m/z) M+H= 543.1 Synthesis of Compound 128:
F
Br 0 F
0 =
ii, 1\1I)-LOH
NC / Cul, Cs2CO3 0 lei 0 HC(OMe)3 L
m H2N * 0 \ __________ = .,,,-., NH3 / \ N
32-f N-- 1 '` N
N ' 1 \
.) 40 ,--cAH L 1 ,N OH
N\
\ , 44-d N 59-a Compound 128 H2N ?---Scheme 59 Step 1: Intermediate 59-a To a solution of intermediate 44-d (261 mg, 0.85 mmol) and intermediate 32-f (285 mg, 0.85 mmol) in 1,4-dioxane (2.8 ml) were sequentially added N,N-dimethylglycine (198 mg, 1.92 mmol), copper(I) iodide (122 mg, 0.64 mmol) and cesium carbonate (1.11 g, 3.42 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate. The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure to provide Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided intermediate 59-a.HCI as a beige foam.
Step 2: Compound 128 Intermediate 59-a (405 mg, 0.85 mmol) and trimethyl orthoformate (6.08 ml, 55.6 mmol) were heated at 110 C for 1 hour. Excess trimethyl orthoformate was removed in vacuo and the residue was treated with 7.0 N
ammonia in Me0H (6.11 ml, 42.8 mmol). The mixture was stirred at room temperature for 3 days and volatiles were removed under reduced pressure.
Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 128.2HCI as a white solid. MS
(m/z) M+H= 501.1 Synthesis of Compound 78:
F
Br 1 0 F
Nj=LOH 0 NC 11 Cul, Cs2CO3 o Si o HC(OMe)3 OTh---\-_______________________________________________ r \ *
Ly---\N NH3 NI-12 . S7/ N
H2N /N-N 29-i 40 S--c \
1, 1 ,N
NC\ N N N\
, N
44-d H2N ------ 60-a Compound 78 Scheme 60 Step 1: Intermediate 60-a To a solution of intermediate 44-d (300 mg, 0.98 mmol) and intermediate 29-i (259 mg, 1.08 mmol) in 1,4-dioxane (3.9 ml) were sequentially added N,N-dimethylglycine (304 mg, 2.95 mmol), copper(I) iodide (187 mg, 0.98 mmol) and cesium carbonate (961 mg, 2.95 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate.
The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided intermediate 60-a=HCI as a beige foam.
Step 2: Compound 78 Intermediate 60-a=HCI (265 mg, 0.57 mmol) and trimethyl orthoformate (1.87 ml, 17.16 mmol) were heated at 110 C for 1 hour. Excess trimethyl orthoformate was removed in vacuo and the residue was treated with 7.0 N
ammonia in Me0H (5.7 ml, 11.44 mmol). The mixture was stirred at room temperature overnight and volatiles were removed under reduced pressure.
Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 78.2HCI as beige solid. MS (m/z) M+H= 491.2 Synthesis of Compound 58:
F
Br O=
N,)-(OH
NH2 /1 NH2 Os S(/ N
Cul, Cs2CO3 \
' N N)....,.., 29-i N 14)....._, U U
10-a Compound 58 Scheme 61 To a solution of intermediate 10-a (3.96 g, 11.1 mmol) and intermediate 29-i (2.91 g, 12.2 mmol) in 1,4-dioxane (55.3 ml) were sequentially added N,N-dimethylglycine (3.42 g, 33.2 mmol), copper(I) iodide (2.10 g, 11.07 mmol) and cesium carbonate (10.82 g, 33.2 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate. The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 58.2HCI as a beige solid. MS (m/z) M+H= 517.2 Synthesis of Compound 117:
F
F
Br 0 0 .
.Ij=(OH OTh"----\-0 el 0 NC Cul, Cs2CO3 HC(OMe)3 / \k, _____________ 1- 0 NH2 it ____________________________________ s,,,N
H2N N.- s__c NH3 \
29-i N -- \
NC N NI N'IN1 OH \ , N \_.4......
H2N N\.___1( OH
48-c OH 62-a Compound 117 Scheme 62 Step 1: Intermediate 62-a To a solution of intermediate 48-c (200 mg, 0.59 mmol) and intermediate 29-1 (157 mg, 0.65 mmol) in 1,4-dioxane (1.50 ml) were sequentially added N,N-dimethylglycine (92 mg, 0.89 mmol), copper(I) iodide (57 mg, 0.29 mmol) and cesium carbonate (583 mg, 1.79 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate.
The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided intermediate 62-a=HCI as a beige solid.
Step 2: Compound 117 Intermediate 62-a=HCI (322 mg, 0.65 mmol) and trimethyl orthoformate (3.0 ml, 19.57 mmol) were heated at 110 C for 1 hour. Excess trimethyl orthoformate was removed in vacuo and the residue was treated with ammonia (7.0 N in Me0H) (1.85 ml, 13.0 mmol). The mixture was stirred at room temperature overnight and volatiles were removed under reduced pressure. Purification by reverse phase chromatography eluting with a i%
aqueous HCl/methanol gradient provided compound 117=2HCI as a white solid. MS (m/z) M+H= 521.1 Synthesis of Compound 100:
F
F
Br 1 0 0 40 0 o=
46, -N 0OH 0 Y\N
S/, N
NC Cul, Cs2CO3 S---c HC(OMe)3 __________________ r _________________________ r-/ \ \
\
, "
H2N N,"m 29-i NC N , nu I , N,-.
OH OH
50-c 63-a Compound 100 Scheme 63 Step 1: Intermediate 63-a To a solution of intermediate 50-c (1.42 g, 4.21 mmol) and intermediate 29-i (1.10 g, 4.63 mmol) in 1,4-dioxane (16.8 ml) were sequentially added N,N-dimethylglycine (1.30 g, 12.6 mmol), copper(I) iodide (802 mg, 4.21 mmol) and cesium carbonate (4.12 g, 12.63 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate. The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided intermediate 63-a=HCI as a white solid.
Step 2: Compound 100 Intermediate 63-a=HCI (577 mg, 1.16 mmol) and trimethyl orthoformate (3.82 ml, 35.0 mmol) were heated at 110 C for 3 hours. Excess trimethyl orthoformate was removed in vacuo and the residue was treated with 7.0 N
ammonia in Me0H (3.30 ml, 23.32 mmol). The mixture was stirred at room temperature overnight and volatiles were removed under reduced pressure.
Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 100.2HCI as white solid. MS
(m/z) M+H= 523.2 Synthesis of Compound 113:
0 0 4, 0 ,49 s.õN NH2 sõNi NH2 =41k $,//N
Bu Tsa N N N\,11 N \ N IsLN \.N
OH 0, 0 0 Compound 100 ti\(. 64-a Compound 113 Scheme 64 Step 1: Intermediate 64-a To a solution of compound 100 (198.0 mg, 0.38 mmol) in THF cooled to -C was slowly added a 2.5 M solution of n-butyllithium in THF (304 pl, 0.76 mmol). After stirring for 30 minutes, para-toluenesulfonyl chloride (72.0 mg, 0.38 mmol) in THF (2 ml) was added. The reaction was stirred at 60 C
overnight and then cooled to room temperature. Water and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 64-a as beige foam.
Step 2: Compound 113 To a solution of intermediate 64-a (85.0 mg, 0.12 mmol) in THF cooled to -10 C was slowly added a 2.5 M solution of n-butyllithium in THF (110.0 pl, 0.27 mmol). The reaction was stirred at 60 C overnight and then cooled to room temperature. Water and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided compound 113 as pale yellow foam. MS (m/z) M+H= 505.2 Synthesis of Compound 125:
Br 0 o,0 4.
Cul, Cs2C0 NC 3 HC(OMe)3 S,/,1%1 I \ N I \,N NH3 N".
29-i I ,N
oI N NL7C0 51-a 65-a Compound 125 Scheme 65 Step 1: Intermediate 65-a To a solution of intermediate 51-a (300 mg, 0.86 mmol) and intermediate 29-i (262 mg, 0.95 mmol) in 1,4-dioxane (1.50 ml) were sequentially added N,N-dimethylglycine (267 mg, 2.59 mmol), copper(I) iodide (165 mg, 0.86 mmol) and cesium carbonate (845 mg, 2.59 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate.
The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 65-a as a beige foam.
Step 2: Compound 125 Intermediate 65-a (120 mg, 0.23 mmol), trimethyl orthoformate (260 pl, 2.3 mmol) and PTSA (catalytic) were stirred at room temperature for 1 hour.
Excess trimethyl orthoformate was removed in vacuo and the residue was treated with 7.0 N ammonia in Me0H (652 pl, 4.6 mmol). The mixture was stirred at room temperature for 3 days and volatiles were removed under reduced pressure. Purification by silica gel chromatography provided compound 125 as a white solid. MS (m/z) M+H= 533.2 Synthesis of Compound 54:
Br ')'OH
N Cul, Cs2CO3 I
NN
N NI)Th 29-i N )ThN
17-a Compound 54 Scheme 66 To a solution of intermediate 17-a (1.20 g, 3.21 mmol) and intermediate 29-1 (844 mg, 3.53 mmol) in DMF (16.0 ml) were sequentially added N,N-dimethylglycine (992 mg, 9.62 mmol), copper(I) iodide (611 mg, 3.21 mmol) and cesium carbonate (4.18 g, 12.83 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate. The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 54.2HCI as a yellow solid. MS (m/z) M+H= 533.1 Synthesis of Compound 55:
F F
Br I V 0 . o=
411 -'NOH
* 0 0 NC Cul, Cs2CO3 =:---\¨ formamidine NH2 / \ ______________ ' NC Si/14 ----0-H2N NN 29-1 " \
.._ 52-a 67-a Compound 55 Scheme 67 Step 1: Intermediate 67-a To a solution of intermediate 52-a (397.0 mg, 1.00 mmol) and intermediate 29-1 (268 mg, 1.12 mmol) in 1,4-dioxane (5.0 ml) were sequentially added N,N-dimethylglycine (157 mg, 1.53 mmol), copper(I) iodide (97.0 mg, 050 mmol) and cesium carbonate (995 mg, 3.05 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate.
The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure to provide intermediate 67-a as beige foam.
Step 2: Compound 55 Formamide (2.84 ml, 71.3 mmol) was added to intermediate 67-a (559 mg, 1.0 mmol) and the reaction was stirred at 180 C for 2 hours. Water and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure.
Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 55.3HCI as beige solid. MS (m/z) M+H= 576.2 Synthesis of Compound 52:
F
Br OH
NH2 41k 441k ,---"\--N \ N 0 NH2 * SerNI
I' N N\ Cs2CO3, Cul I N I
---1 29-1 N rµl_ N--\ ---1 53-a (...__ ) (N--) Compound 52 LO
Scheme 68 To a solution of intermediate 53-a (1.0 g, 2.48 mmol) and intermediate 29-i (771 mg, 3.22 mmol) in DMAC (12.4 ml) were sequentially added quinolin-8-ol (36.0 mg, 0.24 mmol), copper(I) iodide (47.0 mg, 0.24 mmol) and cesium carbonate (808 mg, 2.48 mmol) and the reaction was heated at 140 C for 2 hours. A saturated aqueous solution of ammonium chloride and ethyl acetate were added, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate. The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 52.3HCI as white solid. MS (m/z) M+H= 562.1 Synthesis of Compound 57:
F F
Br i 0 0* 0=
11 fh NC / H NC l formamide * --'----\
, Cr¨ \ NH2 N ____________________________________________ p.
1 CulCs2CO3 H2N N=N 29-i I \,N S(/
14-- . \
i ,N I
H2N N, '---1 N NI, µ----1 --J
54-a 69-a Compound 57 Scheme 69 Step 1: Intermediate 69-a To a solution of intermediate 54-a (425.0 mg, 1.18 mmol) and intermediate 29-1 (282 mg, 1.18 mmol) in 1,4-dioxane (5.9 ml) were sequentially added N,N-dimethylglycine (182 mg, 1.77 mmol), copper(I) iodide (112 mg, 0.59 mmol) and cesium carbonate (1.15 g, 3.54 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate. The combined organic extracts were washed with brine, dried over MgS0.4, filtered and concentrated under reduced pressure to provide intermediate 69-a as a beige foam.
Step 2: Compound 57 Formamide (5.53 ml, 139.0 mmol) was added to intermediate 69-a (600 mg, 1.15 mmol) and the reaction was stirred at 180 C for 4 hours. Water and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure.
Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 57.3HCI as beige solid. MS (m/z) M+H= 546.2 Synthesis of Compound 102:
Br O 0 0 *
LrN
NC Cul, Cs2CO3 I 1 ts( HC(OMe)3 NH, N
H2N N=N 30-b NC NH3 I ,N
44-d 70-a Compound 102 Scheme 70 Step 1: Intermediate 70-a To a solution of intermediate 44-d (4.00 g, 13.1 mmol) and intermediate 30-b (3.38 g, 14.4 mmol) in 1,4-dioxane (43.7 ml) were sequentially added N,N-dimethylglycine (3.04 g, 29.5 mmol), copper(I) iodide (1.87 g, 9.83 mmol) and cesium carbonate (17.08 g, 52.4 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate.
The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided intermediate 70-a=HCI as beige foam.
Step 2: Compound 102 Intermediate 70-a=HCI (3.80 g, 7.68 mmol) and trimethyl orthoformate (54.6 ml, 499.0 mmol) were heated at 110 C for 1 hour. Excess trimethyl orthoformate was removed in vacuo and the residue was treated with 7.0 N
ammonia in Me0H (54.8 ml, 384.0 mmol). The mixture was stirred at room temperature for 2 days and volatiles were removed under reduced pressure.
Purification by reverse phase chromatography eluting with a 10/0 aqueous HCl/methanol gradient provided compound 102.2HCI as white solid. MS
(m/z) M+H= 486.2 Synthesis of Compound 129:
F F
Br 1 0 0 0 o o =
-N o 2qj-OH L' NC = Cul, Cs2CO3 0 I I
Thµl HC(OMe)3 NH2 ik /--N
___________________ y __...
i\ m N , \ N--:"--c \
H2N Ny¨ 30-b NC ,,, ,' NI-13 is I ,N
N
N N, /-'-. H2N .-- i\---45-a 71-a Compound 129 Scheme 71 Step 1: Intermediate 71-a To a solution of intermediate 45-a (350 mg, 1.10 mmol) and intermediate 30-b (283 mg, 1.20 mmol) in 1,4-dioxane (4.3 ml) were sequentially added N,N-dimethylglycine (1.07 g, 3.29 mmol), copper(I) iodide (209 mg, 1.10 mmol) and cesium carbonate (985 mg, 3.02 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate.
The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided intermediate 71-a=HCI as a beige solid.
Step 2: Compound 129 Intermediate 71-a=HCI (403.0 mg, 0.85 mmol) and trimethyl orthoformate (6.07 ml, 55.5 mmol) were heated at 110 C for 3 hours. Excess trimethyl orthoformate was removed in vacuo and the residue was treated with 7.0 N
ammonia in Me0H (6.07 ml, 42.7 mmol). The mixture was stirred at room temperature overnight and volatiles were removed under reduced pressure.
Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 129.2HCI as beige solid. MS
(m/z) M+H= 500.1 Synthesis of Compound 106:
F F
Br 1 0 0 Si 0 0*
,4 '-)LOH . 0 L'N
NC it cui,c.2.3 tst HC(OMe)3 NH2 _________________________________________________ /
/ N N \ NI-----.- 30-b NC N N NH3 \ N1_ l ,N
INI___, /c H2N
43-e 72-a Compound 106 Scheme 72 Step 1: Intermediate 72-a To a solution of intermediate 43-e (350 mg, 1.00 mmol) and intermediate 30-b (260 mg, 1.10 mmol) in 1,4-dioxane (4.0 ml) were sequentially added N,N-dimethylglycine (312 mg, 3.02 mmol), copper(I) iodide (192 mg, 1.00 mmol) and cesium carbonate (985 mg, 3.02 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate.
The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided intermediate 72-a.HCI as a beige solid.
Step 2: Compound 106 Intermediate 72-a=HCI (263 mg, 0.52 mmol) and trimethyl orthoformate (1.72 ml, 15.8 mmol) were heated at 110 C for 2 hours. Excess trimethyl orthoformate was removed in vacuo and the residue was treated with 7.0 N
ammonia in Me0H (46.2 ml, 324 mmol). The mixture was stirred at room temperature overnight and volatiles were removed under reduced pressure.
Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 106.2HCI as a beige solid. MS
(m/z) M+H= 528.1 Synthesis of Compound 114:
Br 9 0 el 0 0*
N,2cOH 0 trN
NC Cul, Cs2CO3 110 I I
HC(OMe)3 NH2 fk N \
H2N /N-N 30-b NC N
OH A-OH
49-c 73-a Compound 114 Scheme 73 Step 1: Intermediate 73-a To a solution of intermediate 49-c (250 mg, 0.78 mmol) and intermediate 30-b (201 mg, 0.85 mmol) in 1,4-dioxane (3.1 ml) were sequentially added N,N-dimethylglycine (241 mg, 2.33 mmol), copper(I) iodide (148 mg, 0.78 mmol) and cesium carbonate (761 mg, 2.33 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate.
The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided intermediate 73-a=FICI as a beige solid.
Step 2: Compound 114 Intermediate 73-a=FICI (106 mg, 0.22 mmol) and trimethyl orthoformate (737 pl, 6.74 mmol) were heated at 110 C for 3 hours. Excess trimethyl orthoformate was removed in vacuo and the residue was treated with 7.0 N
ammonia in Me0H (2.24 ml, 4.49 mmol). The mixture was stirred at room temperature overnight and volatiles were removed under reduced pressure.
Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 114.2HCI as a beige solid. MS
(m/z) M+H= 502.2 Synthesis of Compound 130:
F
F
Br 1 0 0 lei 0 0 =
Nji,.OH0 \
H2N Irl NC 11 Cul, Cs2CO3 f*1 HC(OMe)3 NH240 N---;"-k 'N NC N
\ OH \ 30-b NH3 Isr" \
' N
, N I ,N
rJ0H H2N Z-----/
OH ( OH OH
50-c 74-a Compound 130 Scheme 74 Step 1: Intermediate 74-a To a solution of intermediate 50-c (300 mg, 0.89 mmol) and intermediate 30-b (229 mg, 0.97 mmol) in 1,4-dioxane (890 pl) were sequentially added N,N-dimethylglycine (275 mg, 2.67 mmol), copper(I) iodide (169 mg, 0.89 mmol) and cesium carbonate (870 mg, 2.67 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate.
The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided intermediate 74-a=HCI as beige solid.
Step 2: Compound 130 A solution of intermediate 74-a (125 mg, 0.25 mmol), trimethyl orthoformate (836 pl, 7.65 mmol) and PTSA (catalytic) was stirred at room temperature for 3 hours. Excess trimethyl orthoformate was removed in vacuo and the residue was treated with ammonia (7.0 N in Me0H) (2.55 ml, 5.10 mmol).
The mixture was stirred at room temperature for 2 days and volatiles were removed under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 130.2HCI as a white solid. MS (m/z) M+H= 518.1 Synthesis of Compound 97:
F F
fa Br 1 ?I O= o, it --=.-\ 0 ----\-NC Cul, Cs2CO3 HC(OMe)3 NH2 iii -.........*. C _____________________ I.
/ \ \/N , \ /
H2N V'm 36-f I ,N CNNo I NH3 ,N Co N N
)\ HN2N N)Th N
U
0 0 \--0) irk 43-e 75-a Compound 97 Scheme 75 Step 1: Intermediate 75-a To a solution of intermediate 43-e (101 mg, 0.29 mmol) and intermediate 36-f (110 mg, 0.32 mmol) in 1,4-dioxane (1.4 ml) were sequentially added N,N-dimethylglycine (90 mg, 0.87 mmol), copper(I) iodide (56 mg, 0.29 mmol) and cesium carbonate (381 mg, 1.17 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate.
The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 75-a as a beige solid.
Step 2: Compound 97 Intermediate 75-a (160 mg, 0.26 mmol), trimethyl orthoformate (1.86 ml, 17.09 mmol) and TFA (catalytic) were heated at 110 C for 1 hour. Excess trimethyl orthoformate was removed in vacuo and the residue was treated with 7.0 N ammonia in Me0H (1.87 ml, 13.14 mmol). The mixture was stirred at 50 C overnight and volatiles were removed under reduced pressure. Purification by reverse phase chromatography eluting with a 1 70 aqueous HCl/methanol gradient provided compound 97.2HCI as a beige solid.
MS (m/z) M+H= 636.1 Synthesis of Compound 99:
F F
0 46 0 .
NH2 OH2 Pd/C NH2 fit ---=--\-_____________________________________ ).
NV r "N N ,N NV i 1 "N N N
I ,) o a * 0 Compound 97 Compound 99 Scheme 76 A solution of compound 97.2HCI (100 mg, 0.15 mmol) in ethyl acetate was treated with 10% palladium on carbon (32 mg, 0.015 mmol) and purged with H2. The solution was stirred under H2(1 atm) for 1 hour before being filtered through celite. The filtrate was concentrated in vacuo. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 99.2HCI as white solid. MS (m/z) M+H= 546.2 Synthesis of Compound 93:
F F
Br 1 9 0 * o 0 *
NJ-OH
NC 41 Cul, Cs2CO3 4. -------% *
___________________ w NC N - + NC N.--,,/N---/
H2N N-- 37-f + 37-f 1 \,N r , \,N
H2N )_____\ 1 N
\-SiMe3 -0) 43-e 77-a 77-a' F
0.
F
0 ft HC(OMe)3 NH2 * -":---"\--- NH2 *
77-a + 77-a' --1.- N / N + , -_/
NH3 N "*" i " N N
L 1 N r -z N ' \
l'i, o N' SiMe3 77-h 77-h' F
O, HCI NH2 * 0 ---\-- õ, 77-h + 77-b' ---0,- / HNrµ
N
l " ,N
N l'I)_Th U
Compound 93 Scheme 77 Step 1: Intermediates 77-a and 77-a' To a solution of intermediate 43-e (115 mg, 0.33 mmol) and intermediates 37-f and 37-f' (123 mg, 0.36 mmol) in 1,4-dioxane (1.6 ml) were sequentially added N,N-dimethylglycine (102 mg, 0.99 mmol), copper(I) iodide (63 mg, 0.33 mmol) and cesium carbonate (431 mg, 1.32 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate. The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 77-a and 77-a' as an inseparable mixture.
Step 2: Intermediates 77-b and 77-b' A solution of intermediates 77-a and 77-a' (100 mg, 0.16 mmol), trimethyl orthoformate (2.0 ml, 18.28 mmol) and PTSA (catalytic) was stirred at room temperature for 1 hour. Excess trimethyl orthoformate was removed in vacuo and the residue was treated with 7.0 N ammonia in Me0H (2.0 ml, 14.1 mmol). The mixture was stirred at room temperature for 3 days and volatiles were removed under reduced pressure. Purification by silica gel chromatography provided intermediates 77-b and 77-b' as an inseparable mixture.
Step 3: Compound 93 4N HCI in 1,4-dioxane (2 mL) was added to intermediates 77-b and 77-b' (60 mg, 0.09 mmol) band the mixture was stirred for 1 hour at room temperature. Volatiles were removed under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 93.2HCI as a white solid. MS (m/z) M+H= 502.2 Synthesis of Compound 118:
F F
Br 1 On 0 * 0 .
NC 11 Cut, Cs2CO3 *--------\-- *
/ \
___________________ ).
N NC r-.N m e'' + NC \
I
H2N N-- 38-e +38-e' 1 ",N 1 1 N
a N 0 H2N __ )__\ 1 SiMe3 N
\
0 --0) \---0) 43-e 78-a 78-a' F F
0 10 0 .
HC(OMe)3 NH2 4* -----\--- n, + NH2 4* ,---=\-(:)/"SiMe3 78-a + 78-a' 0- N r II N.--/
NH3 N ' , \
r '( N" \
I N'N NK \
N Q t`l..Th I N
SiMe3 78-b 78-b' F
0 .
HCI NH2 . ------'\
78-b + 78-b' 1. ' HN ,,, N
N \
Th\I r4)____I
Compound 118 Scheme 78 Step 1: Intermediates 78-a and 78-a' To a solution of intermediate 43-e (276.0 mg, 0.79 mmol) and intermediates 38-e and 38-e' (340 mg, 0.87 mmol) in 1,4-dioxane (795 pl) were sequentially added N,N-dimethylglycine (246 mg, 2.38 mmol), copper(I) iodide (151 mg, 0.79 mmol) and cesium carbonate (1.03 g, 3.18 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate. The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 78-a and 78-a' as an inseparable mixture.
Step 2: Intermediates 78-b and 78-b' A solution of intermediates 78-a and 78-a' (90 mg, 0.14 mmol), trimethyl orthoformate (1.03 ml, 9.45 mmol) and TFA (catalytic) was stirred at room temperature for 1 hour. Excess trimethyl orthoformate was removed in vacuo and the residue was treated with 7.0 N ammonia in Me0H (1.03 ml, 7.27 mmol). The mixture was stirred at room temperature overnight and volatiles were removed under reduced pressure to provide intermediates 78-b and 78-b' as an inseparable mixture.
Step 3: Compound 118 4N HCI in 1,4-dioxane (2.82 mL) was added to intermediates 78-b and 78-b' (60 mg, 0.09 mmol) and the mixture was stirred overnight at room temperature. Volatiles were removed under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 118.2HCI as a white solid. MS (m/z) M+H=
516.2 Synthesis of Compound 90:
F F
Br 1 ?I 0O 0 fa 41, NC Cul, Cs2CO3 0HC(OMe)3 NH2 * ---\----/ \
NC ,N,.// NH3 N ''. \
H2N N'N 34-d 1 \ N L I N
H N
H2N N\
M
OH OH OH
47-a 79-a Compound 90 Scheme 79 Step 1: Intermediate 79-a To a solution of intermediate 79-a (310 mg, 1.0 mmol) and intermediate 34-d (313 mg, 1.21 mmol) in 1,4-dioxane (2.5 ml) were sequentially added N,N-dimethylglycine (156 mg, 1.51 mmol), copper(I) iodide (96.0 mg, 0.50 mmol) and cesium carbonate (1.31 g, 4.04 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate. The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided intermediate 79-a as a beige solid.
Step 2: Compound 90 Intermediate 79-a (280 mg, 0.62 mmol) and trimethyl orthoformate (2.05 ml, 18.7 mmol) were heated at 110 C for 3 hours. Excess trimethyl orthoformate was removed in vacuo and the residue was treated with 7.0 N
ammonia in Me0H (1.78 ml, 12.5 mmol). The mixture was stirred at room temperature overnight and volatiles were removed under reduced pressure.
Purification by reverse phase chromatography eluting with a 1 /0 aqueous HCl/methanol gradient provided compound 90.2HCI as a beige solid. MS
(m/z) M+H= 476.2 Synthesis of Compound 103:
F F
Br 1 0 0 * AO o O*
NC Cul, Cs2CO3 fl Cr---:\ HC(OMe)3 NH2 . -.---;--\,, _________________________________________________ s N / N
/ \ NC ,...- ---./ NH3 N , \
H2N N'N 34-d =
1 N L I ,N
H2N o ,N N)._..., U
46-a 80-a Compound 103 Scheme 80 Step 1: Intermediate 80-a To a solution of intermediate 46-a (300 mg, 0.87 mmol) and intermediate 34-d (247 mg, 0.95 mmol) in 1,4-dioxane (2.1 ml) were sequentially added N,N-dimethylglycine (134 mg, 1.30 mmol), copper(I) iodide (83 mg, 0.43 mmol) and cesium carbonate (1.13 g, 3.48 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate. The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided intermediate 80-a=FICI as a beige solid.
Step 2: Compound 103 Intermediate 80-a=HCI (65.0 mg, 0.13 mmol) and trimethyl orthoformate(3.0 ml, 4.01 mmol) were heated at 110 C for 3 hours. Excess trimethyl orthoformate was removed in vacuo and the residue was treated with 7.0 N
ammonia in Me0H (378 pl, 2.65 mmol). The mixture was stirred at room temperature overnight and volatiles were removed under reduced pressure.
Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 103.2HCI as a white solid. MS
(m/z) M+H= 514.2 Synthesis of Compound 120:
Br O * 410 o *
NC Cul, Cs2003 = HC(OMe)3 NH2 10 N
N
HN N NC NH3'N 35-d i I JA
\,N1 42-d 81-a Compound 120 Scheme 81 Step 1: Intermediate 81-a To a solution of intermediate 42-d (176 mg, 0.53 mmol) and intermediate 35-d (145 mg, 0.53 mmol) in 1,4-dioxane (3.5 ml) were sequentially added N,N-dimethylglycine (123 mg, 1.19 mmol), copper(I) iodide (76 mg, 0.40 mmol) and cesium carbonate (693 mg, 2.13 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated, and the aqueous phase was extracted twice with ethyl acetate.
The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided intermediate 81-a.HCI as a yellow foam.
Step 2: Compound 120 Intermediate 81-a=HCI (259 mg, 0.53 mmol) and trimethyl orthoformate (3.78 ml, 34.6 mmol) were heated at 110 C for 1 hour. Excess trimethyl orthoformate was removed in vacuo and the residue was treated with 7.0 N
ammonia in Me0H (3.80 ml, 26.6 mmol). The mixture was heated at 60 C
for 5 hours and volatiles were removed under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 120.2HCI as a white solid. MS (m/z) M+H=
514.2 Synthesis of intermediate 82-c:
Br Br Br 11 0 DIPEA =
TMS-diazomethane =CI malononitrile CN CN
HO Me0 CN CN
82-a 82-b 82-c Scheme 82 Step 1: Intermediate 82-b To a solution of 4-bromo-2-fluorobenzoyl chloride (16.27 g, 68.5 mmol) in toluene (85 ml) and THF (8.5 ml), cooled to -10 C, were sequentially added malononitrile (4.75 g, 71.9 mmol) and DIPEA (23.93 ml, 137.0 mmol) in toluene (25 mL) drop wise over a period of 1 hour. After the addition was completed, the reaction was stirred for 2 hours at 0 C and room temperature for an additional 2 hours. Volatiles were removed under reduced pressure.
1N HCI and ethyl acetate were added to the residue; the organic layer was separated, washed twice with 1N HCI and brine, dried over MgSO4, filtered and concentrated under reduced pressure to provide intermediate 82-b as a yellow solid.
=
Step 2: Intermediate 82-c To a solution of intermediate 82-b (18.29 g, 68.5 mmol) in acetonitrile (247 ml) and methanol (27.4 ml), cooled to 0 C, was added DIPEA (14.36 ml, 82.0 mmol) and a 2M solution of diazomethyl)trimethylsilane in hexanes (37.7 ml, 75.0 mmol). After the addition was completed, the reaction was stirred at room temperature overnight. Acetic acid (1.17 ml, 20.5 mmol) was added, the reaction was stirred for 30 minutes and volatiles were removed under reduced pressure. A saturated aqueous solution of NaHCO3 and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure.
Purification by silica gel chromatography provided intermediate 82-c as a yellow solid.
Synthesis of Intermediate 83-a:
Br Br TEA
441k NC
NC 43-d 0 \ N
82-c 83-a Scheme 83 To a solution of intermediate 82-c (2.0 g, 7.12 mmol) and TEA (1.98 ml, 14.23 mmol) in Et0H (3.50 ml) was added intermediate 43-cl=FICI (1.30 g, 8.54 mmol) and the reaction was then stirred for 2 hours at 100 C. Volatiles were removed under reduced pressure. A saturated aqueous solution of ammonium chloride and ethyl acetate were added to the residue, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure to provide intermediate 83-a as a yellow solid.
Synthesis of Compound 60:
Br 0 I. 0 0 =
NH2 41k s,õN
NC F Cul, Cs2CO3 Formamide I "'N NC N N
N 1_ 29-i 83-a 84-a Compound 60 Scheme 84 Step 1: Intermediate 84-a To a solution of intermediate 83-a (2.60 g, 7.12 mmol) and intermediate 29-1 (1.00 g, 4.18 mmol) in 1,4-dioxane (20.9 ml) were sequentially added N,N-dimethylglycine (646 mg, 6.27 mmol), copper(I) iodide (398 mg, 2.09 mmol) and cesium carbonate (4.09 g, 12.5 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated, and the aqueous phase was extracted twice with ethyl acetate. The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 84-a as a beige solid.
Step 2: Compound 60 Formamide (11.7 ml, 293 mmol) was added to intermediate 84-a (2.18 g, 4.18 mmol) and the reaction was stirred at 180 C overnight. Water and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure.
Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 60.2HCI as white solid. MS (m/z) M+H= 551.2 Synthesis of Compound 73:
F
F
Br 0 el 0 0 =
F /N---z/ NH2 . --)---\
1\1 N
NC
Cul, Cs2CO3 HC(OMe)3 F F
_________________ ).
I N m ' _________________ N1LT
\ NC N
1 \,N
\ Nr NH3 H2N N 34-d y..,.\ H2N N
(--()I
83-a 85-a Compound 73 Scheme 85 Step 1: Intermediate 85-a To a solution of intermediate 83-a (2.60 g, 7.12 mmol) and intermediate 34-d (1.0 g, 4.18 mmol) in 1,4-dioxane (20.9 ml) were sequentially added N,N-dimethylglycine (646 mg, 6.27 mmol), copper(I) iodide (398 mg, 2.09 mmol) and cesium carbonate (4.09 g, 12.5 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated, and the aqueous phase was extracted twice with ethyl acetate. The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 85-a as a beige solid.
Step 2: Compound 73 Intermediate 85-a (195.0 mg, 0.38 mmol) and trimethyl orthoformate (1.26 ml, 11.59 mmol) were heated at 110 C for 3 hours. Excess trimethyl orthoformate was removed in vacuo and the residue was treated with 7.0 N
ammonia in Me0H (3.86 ml, 7.72 mmol). The mixture was stirred at room temperature overnight and volatiles were removed under reduced pressure.
Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 73.HCI as white solid. MS (m/z) M+H= 534.1 Synthesis of Compound 95:
F
F
Br 0 el 0 0 .
4k ,NIOH 40 1 N
/N
NC F Cul, Cs2CO3 __________________ > F ., HC(OMe)3 __________________________________________________ N . O_:21 " F
I \, H2N N NC ,,, 1 31-d \ Nr. [,,, ,N
N
11)____, (--)1, 83-a 86-a Compound 95 Scheme 86 Step 1: Intermediate 86-a To a solution of intermediate 83-a (200 mg, 0.54 mmol) and intermediate 31-d (177 mg, 0.65 mmol) in 1,4-dioxane (3.65 ml) were sequentially added N,N-dimethylglycine (127 mg, 1.23 mmol), copper(I) iodide (7.8 mg, 0.41 mmol) and cesium carbonate (714 mg, 2.19 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated, and the aqueous phase was extracted twice with ethyl acetate.
The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided intermediate 86-a=HCI as a beige solid.
Step 2: Compound 95 Intermediate 86-a=HCI (284 mg, 0.54 mmol) and trimethyl orthoformate (3.90 ml, 35.6 mmol) were heated at 110 C for 1 hour. Excess trimethyl orthoformate was removed in vacuo and the residue was treated with 7.0 N
ammonia in Me0H (3.91 ml, 27.4 mmol). The mixture was stirred at room temperature overnight and volatiles were removed under reduced pressure.
Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 95.2HCI as white solid. MS (m/z) M+H= 545.2 Synthesis of Intermediate 87-a:
Br Br ,.
F NC F
NC 42-c 0 I \ N
82-c 87-a a Scheme 87 To a solution of intermediate 82-c (1.00 g, 3.56 mmol) and TEA (1.09 ml, 7.83 mmol) in Et0H (3.50 ml) was added intermediate 42-c (583 mg, 4.27 mmol) and the reaction was then stirred for 2 hours at 100 C. Volatiles were removed under reduced pressure. A saturated aqueous solution of ammonium chloride and ethyl acetate were added to the residue, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 87-a as beige solid.
Synthesis of Compound 92:
F
F
Br 0 el 0 0 =
. ''N -)LOH 40 r\,N
/N--Zi NH2 10 NC F Cul, Cs2CO3 F
_________________ 1.- HC(OMe)3 F
NC "
I \,N K1 NH3 34-d \ Nr L I pi H2N f \l,..,_ N NL
c-J H2N 0 U
87-a 88-a Compound 92 Scheme 88 Step 1: Intermediate 88-a To a solution of intermediate 87-a (452 mg, 1.29 mmol) and intermediate 34-d (368 mg, 1.42 mmol) in 1,4-dioxane (5.20 ml) were sequentially added N,N-dimethylglycine (400 mg, 3.88 mmol), copper(I) iodide (247 mg, 1.29 mmol) and cesium carbonate (1.26 g, 3.88 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated, and the aqueous phase was extracted twice with ethyl acetate. The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided intermediate 88-a=HCI as a beige solid.
Step 2: Compound 92 Intermediate 88-a=HCI (124.0 mg, 0.25 mmol) and trimethyl orthoformate (935 pi, 7.63 mmol) were heated at 110 C for 3 hours. Excess trimethyl orthoformate was removed in vacuo and the residue was treated with 7.0 N
ammonia in Me0H (2.54 ml, 5.09 mmol). The mixture was stirred at room temperature overnight and volatiles were removed under reduced pressure.
Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 92.2HCI as a beige solid. MS
(m/z) M+H= 518.2 Synthesis of intermediate 89-a:
Br Br _____________________________________________________ , *
F TEA NC F
NC 44-c 0 l \ N
)----82-c 89-a Scheme 89 To a solution of intermediate 82-c (2.0 g, 7.12 mmol) and TEA (2.18 ml, 15.65 mmol) in Et0H (7.12 ml) was added intermediate isopropylhydrazine hydrochloride (944 mg, 8.54 mmol) and the reaction was then stirred for 2 hours at 100 C. Volatiles were removed under reduced pressure. A
saturated aqueous solution of ammonium chloride and ethyl acetate were added to the residue, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure.
Purification by silica gel chromatography provided intermediate 89-a as a beige solid.
Synthesis of Compound 98:
F
F
Br 0OO 0 =
iii .--14).1''ON 40 Cr\,N
F
F /N --S HC(OMe)3 NH2 . .----\-- m NC Cul, Cs2CO3 , N F
l \,N NC õI 1 \ NH3 i_ ,N
34-d N :. N\
H2N Nj H2N---1."--89-a 90-a Compound 98 Scheme 90 Step 1: Intermediate 90-a To a solution of intermediate 89-a (400 mg, 1.24 mmol) and intermediate 34-d (352 mg, 1.36 mmol) in 1,4-dioxane (4.9 ml) were sequentially added N,N-dimethylglycine (383 mg, 3.71 mmol), copper(I) iodide (236 mg, 1.24 mmol) and cesium carbonate (1.21 g, 3.71 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated, and the aqueous phase was extracted twice with ethyl acetate. The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided intermediate 90-a=FICI as beige solid.
Step 2: Compound 98 Intermediate 90-a=HCI (208 mg, 0.45 mmol) and trimethyl orthoformate (1.47 ml, 13.4 mmol) were heated at 110 C for 3 hours. Excess trimethyl orthoformate was removed in vacuo and the residue was treated with 7.0 N
ammonia in Me0H (4.48 ml, 8.96 mmol). The mixture was stirred at room temperature for 3 days and volatiles were removed under reduced pressure.
Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 98.2HCI as a beige solid. MS
(m/z) M+H= 492.2 Synthesis of Intermediate 91-d:
=
Br Br Br Br F F is F 0 oxalyl chloride F 0 ________________ ).- malononitrile =TMS-diazomethane NaOH
CN CN
HO '=-= Me0 0 OH 0 Cl CN CN
91-a 91-b 91-c 91-d Scheme 91 Step 1: Intermediate 91-b To a suspension of 4-bromo-3-fluorobenzoic acid (15.0 g, 68.5 mmol) in dichloromethane (342.0 ml) was added DMF (106.0 pl, 1.37 mmol) and oxaly1y1 chloride (8.99 ml, 103.0 mmol). The solution was then stirred at room temperature for 3 hours. Volatiles were removed under reduced pressure to provide intermediate 91-b as a yellow solid.
Step 2: Intermediate 91-c To a solution of intermediate 91-b (16.27 g, 68.5 mmol) in toluene (85.0 ml) and THF (8.5 ml) cooled to -10 C were sequentially added malononitrile (4.75 g, 71.9 mmol) and DIPEA (23.93 ml, 137 mmol) drop wise over a period of 15 minutes. The reaction was stirred at 0 C for 2 hours and room temperature for an additional 2 hours. Volatiles were removed under reduced pressure. Ethyl acetate and 1N HCI were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated in vacuo to provide intermediate 91-c as a yellow solid.
Step 3: Intermediate 91-d To a solution of intermediate 91-c (18.29 g, 68.5 mmol) in acetonitrile (247.0 ml) and Me0H (27.4 ml) cooled to 0 C were sequentially added DIPEA (14.36 ml, 82.0 mmol) and a 2M solution of TMS-Diazomethane in hexanes (37.7 ml, 75.0 mmol). The reaction was then stirred for 5 hours at room temperature. Acetic acid (1.17 ml, 20.55 mmol) was then added and the reaction was stirred for an additional 30 minutes. Volatiles were removed under reduced pressure. A saturated aqueous solution of NaHCO3 and ethyl acetate was added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated in vacuo. Purification by silica gel chromatography provided intermediate 91-d as a beige solid.
Synthesis of Intermediate 92-a:
Br Br F
TEA
NC NC
43-d CN
H2N I \N
91-d 92-a Scheme 92 To a solution of intermediate 91-d (2.0 g, 7.12 mmol) and TEA (1.98 ml, 14.23 mmol) in Et0H (3.50 ml) was added intermediate 43-d.HCI (1.30 g, 8.54 mmol) and the reaction was then stirred for 2 hours at 100 C. Volatiles were removed under reduced pressure. A saturated aqueous solution of ammonium chloride and ethyl acetate were added to the residue, the organic layer was separated, washed with brine, dried over MgSO4, ffitered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 92-a as a beige solid.
Synthesis of Compound 59:
Br 0 I. 0 0 j=LOH
8,711 Cul, Cs2CO3 Formamide NH2 NC
I "'NI NC \"N
__________________________________________________ N
29-i N I ,N
N
-12N N)..Th 92-a 93-a Compound 59 Scheme 93 Step 1: Intermediate 93-a To a solution of intermediate 92-a (2.60 g, 7.12 mmol) and intermediate 29-1 (1.0 g, 4.18 mmol) in 1,4-dioxane (20.90 ml) were sequentially added N,N-dimethylglycine (646 mg, 6.27 mmol), copper(I) iodide (398 mg, 2.09 mmol) and cesium carbonate (4.09 g, 12.54 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated, and the aqueous phase was extracted twice with ethyl acetate. The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 93-a as a beige solid.
Step 2: Compound 59 Formamide (11.67 ml, 293.0 mmol) was added to intermediate 93-a (2.18 g, 4.18 mmol) and the reaction was stirred at 180 C overnight then cooled to room temperature. Water and ethyl acetate were added, the organic layer was separated, washed with brine, dried over Mg504, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 59.2HCI as a yellow solid. MS (m/z) M+H= 551.1 Synthesis of Compound 72:
F
F
Br 0 Si 0 o =
th F -7`0H
N-(/ .- it NH, ,..N N
NC
Cul, Cs2CO3 HC(OMe)3 i "N NC N m 34-d \ NI NH3 N)_Th (---1;) 92-a 94-a Compound 72 Scheme 94 Step 1: Intermediate 94-a To a solution of intermediate 92-a (291.0 mg, 0.79 mmol) and intermediate 34-d (227.0 mg, 0.87 mmol) in 1,4-dioxane (3.2 ml) were sequentially added N,N-dimethylglycine (247.0 mg, 2.39 mmol), copper(I) iodide (152.0 mg, 0.79 mmol) and cesium carbonate (780 mg, 2.39 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated, and the aqueous phase was extracted twice with ethyl acetate.
The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided intermediate 94-a=FICI as a beige solid.
Step 2: Compound 72:
Intermediate 94-a=HCI (56.0 mg, 0.11 mmol) and trimethyl orthoformate (363 pl, 3.32 mmol) were heated at 110 C for 3 hours. Excess trimethyl orthoformate was removed in vacuo and the residue was treated with 7.0 N
ammonia in Me0H (1.10 ml, 2.21 mmol). The mixture was stirred at room temperature overnight and volatiles were removed under reduced pressure.
Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 72.2HCI as a white solid. MS
(m/z) M+H= 534.2 Synthesis of Compound 68:
F
Br 0 411t , 0=
0--......, , \N
NH2 4410 N)-LOH NH2 *
---j N \ Cul, Cs2CO3 N.:
N
l , _____________ 0, N'"N
N N)_\ I , \--__) .
0 39-b N
U
17-a Compound 68 Scheme 95 To a solution of intermediate 17-a (300.0 mg, 0.80 mmol) and intermediate 39-b (177.0 mg, 0.80 mmol) in 1,4-dioxane (3.6 ml) were sequentially added N,N-dimethylglycine (372.0 mg, 3.61 mmol), copper(I) iodide (229.0 mg, 1.20 mmol) and cesium carbonate (1.04 g, 3.21 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated, and the aqueous phase was extracted twice with ethyl acetate.
The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 68.2HCI as a beige solid. MS (m/z) M+H= 514.2 Synthesis of Compound 69:
F
Br 0 .
----)r-N
NH2 * N).LOH NH2 40 N\j,_ N \ N Cul, Cs2CO3 I, \
N N)__\ L I ,N
U0 40-b N
) 17-a Compound 69 Scheme 96 To a solution of intermediate 17-a (300.0 mg, 0.80 mmol) and intermediate 40-b (177.0 mg, 0.80 mmol) in 1,4-dioxane (3.6 ml) were sequentially added N,N-dimethylglycine (372.0 mg, 3.61 mmol), copper(I) iodide (229.0 mg, 1.20 mmol) and cesium carbonate (1.04 g, 3.21 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated, and the aqueous phase was extracted twice with ethyl acetate.
The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 69.2HCI as a beige solid. MS (m/z) M+H= 514.2 Synthesis of Compound 63:
Br NH2 Os0 111j-LOH 0 INV \ Cul, Cs2CO3 1 'N N
N 1 'N
0 41-b N
17-a Compound 63 Scheme 97 A solution of intermediate 17-a (437 mg, 1.17 mmol), intermediate 11-c (255 mg, 1.17 mmol), quinolin-8-ol (34 mg, 0.23 mmol), copper (I) iodide (44.0 mg, 0.23 mmol) and cesium carbonate (761 mg, 2.33 mmol), in dimethylacetamide (1.2 ml), was degassed with argon for 10 minutes, heated in a sealed tube at 140 C overnight and then cooled to room temperature. Water and ethyl acetate were added, the organic layer was separated, the aqueous layer was extracted twice with ethyl acetate, the combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% HCl/methanol gradient provided compound 63.1-1CI as a yellow solid. MS (m/z) M+H= 512.2 Synthesis of Compound 67:
N
i F0 Br K2CO3 F Br Cul, DEA, Pd(PPh3)2Cl2 F 0 NI\
_________________ * __ MOMCI Vi 1 Nr ----_:-----XN1 OH OMOM OMOM
I
98-a 98-b 98-c 98-d N N
F I F i H2 Pd/C N HCI
98-d ______________ )= 1 OMOM OH
98-e 98-f F
Br NH2 41* N / N
..--- -...../
NI ' \N Cul I' N NI)Th . I ,N
98-f N NI)______\
S---.0) U
17-a 0 compound 67 Scheme 98 Step 1: Intermediate 98-b To a solution of 3-bromo-5-fluorophenol, 98-a (25.00 g, 131.0 mmol), in acetone (654 ml) were sequentially added K2CO3 (27.10 g, 196.0 mmol) and chloro(methoxy)methane (11.59 g, 144.0 mmol). The reaction was stirred at room temperature for 2 hours and then filtered. The filtrate was concentrated under reduced pressure to provide intermediate 98-b as a yellow oil.
Step 2: Intermediate 98-d To a solution of intermediate 98-b (2.00 g, 8.51 mmol) in DMF (17.02 ml), were sequentially added diethylamine (975 pl, 9.36 mmol), copper(I)iodide (65 mg, 0.34 mmol) and 5-ethyny1-1-methyl-1H-imidazole 98-c (948 mg, 8.93 mmol). After copper(I)iodide has completely dissolved, Dichlorobis(triphenylphosphine)palladium(II) (119 mg, 0.17 mmol) was added and the reaction was then stirred at 100 C overnight and then cooled to room temperature. Water and ethyl acetate were added, the organic layer was separated, the aqueous layer was extracted twice with ethyl acetate, the combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 98-d as a beige solid Step 3: Intermediate 98-e A solution of intermediate 98-d (600.0 mg, 2.3 mmol) in methanol was treated with 10% palladium on carbon (245.0 mg, 0.01 mmol) and purged with H2. The solution was stirred under H2 (1 atm) overnight before being filtered through celite. The filtrate was concentrated in vacuo to provide intermediate 98-e as a yellow oil.
Step 4: Intermediate 98-f To a solution of intermediate 98-e (2.4 g, 9.08 mmol) in Me0H (17 ml) was added 4N HCI in dioxane (2.76 ml, 91 mmol). The reaction mixture was stirred at room temperature for 2 hours. Volatiles were removed under reduced pressure, diethyl ether was added to the residue; a precipitate formed and was collected by filtration to provide intermediate 98-f=HCI as a white solid.
Step 5: Compound 67 A solution of intermediate 17-a (292 mg, 0.78 mmol), intermediate 98-f=HCI
(200 mg, 0.78 mmol), tetramethylheptane-3,5-dione (287 mg, 1.56 mmol), copper (I) iodide (148 mg, 0.78 mmol) and cesium carbonate (762 mg, 2.33 mmol), in NMP (3.9 ml), was degassed with argon for 10 minutes, heated in a sealed tube at 120 C overnight and then cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated, and the aqueous phase was extracted twice with ethyl acetate. The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% HCl/methanol gradient provided compound 67.2HCI as a yellow solid. MS (m/z) M+H= 514.3 Compounds 7, 8, 11, 13, 14, 19, 21, 24 to 28, 33, 34, 37 to 51, 53, 56, 61, 62, 64, 66, 70, 71, 74-77, 79 to 84, 86 to 89, 94, 96, 104, 105, 107 to 112, 115, 116, 119, 121, 122, 123, 124, 126, 127, 131, 132, and 133 were prepared using similar methods to those described above.
Table 1 summarizes representative compound of Formula 1.
Table 1: Example Compounds of Formula 1 Compound Structure MS (m/z) o, 1 NH2 * [M+H]=410.2 N.-- \ O
1 ,N
N N
H
o, NH2 4It 40 [M+H]=478.2 N' 1 \
I N
N N),õ_, U
o, NH2 *
0[M+H]=593.1 3 N \
L 1 ,N
NN,_____ e 0--.
o, NH2 *
* [M+H]=493.1 N" \N
N N, òH
o, NH2 *
N" * [M+H]=547.1 I \N
' N N
o, NH2 44*
6 fh[M+H]=535.1 N -' \ N
I ' N N
o, NH2 4.
O[M+Hr=563.1 N N
----\
0 =
NH2 4k 8 N '' , \ O [M+H]=563.1 I ,N
N N
L\N---e i----, NH2 4.
=
9 ci [M+H]=512.2 N \
I
N N
o, NH2 it [M+H]=508.1 , \ ¨0 I ,N
N N
*
11 0, [M+H]=508.2 \N
I ' N
o*
CN
NH2 41i =
12 [M+H]=503.2 N' \N
N N
o, NH2. .
13 [M+Hr=478.2 N \N
I ' N NL
U
0 =
NH2 44* S N
14 [M+Hr=485.2 N' \N
I ' U
o, NH2. .
15 [M+Hr=503.3 N' \N
l , N NL
U
o, 16 NH2 0 fb [M+H]=424.2 N' \N
l , N N
\
o, CN
17 NH2 fi fa [M+H]=449.3 N' , \
L 1 ,N
1\1 N
\
o,
overnight. Volatiles were removed in vacuo. Purification by silica gel chromatography provided intermediate 17-a as a white solid.
Synthesis of compound 22:
0 . CN
CuCI, Cs2CO3 17-a + 14-d OH __ ). NH2 44k 4411k N
=NV "
i 1 ,N
N N Compound 22 a Scheme 18 A solution of intermediate 17-a (200 mg, 0.53mmol), intermediate 14-d (181 mg, 0.80 mmol), quinolin-8-ol (15.5 mg, 0.11 mmol), copper (I) chloride (11.5 mg, 0.11 mmol) and cesium carbonate (348 mg, 1.07 mmol) in dimethylacetamide (5.3 ml) was degassed with argon for 10 minutes, heated in a sealed tube at 140 C overnight and then cooled to room temperature.
Water and ethyl acetate were added, the organic layer was separated, the aqueous layer was extracted twice with ethyl acetate, the combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with 1% HCl/methanol gradient provided compound 22=HCI as a beige solid. MS (m/z) M+H= 519.2 Synthesis of compound 31:
Ph3P, DIAD, o TBAF 0 1.1 14-b CF3 OTBS OH
HO SI
CF3 19-a 19-b CuCI, Cs2CO3, 0 12-a + 19-b N \N
I ' / N N ¨
\ Compound 31 Scheme 19 Step 1: Intermediate 19-a To a solution of intermediate 14-b (10.0 g, 41.9 mmol) in THF (210 mL) were sequentially added 2-(trifluoromethyl)phenol (6.80 g, 41.9 mmol), triphenylphosphine (13.2 g, 50.33 mmol) and DIAD (9.79 ml, 50.3 mmol) drop wise at room temperature. The reaction was then stirred at room temperature overnight. Saturated aqueous ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure.
Purification by silica gel chromatography provided intermediate 19-a as a colorless oil.
Step 2: Intermediate 19-b To a solution of intermediate 19-a (13.9 g, 36.3 mmol) in THF (182.0 ml) was added a 1.0M solution of tetrabutylammonium fluoride in THF (36.3 ml, 36.3 mmol) and the reaction was stirred at room temperature for 1 hour.
Saturated aqueous ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 19-b as colorless oil.
Step 3: Compound 31 A solution of intermediate 12-a (200 mg, 0.66 mmol), intermediate 19-b (265 mg, 0.98 mmol), quinolin-8-ol (19.0 mg, 0.13 mmol), copper (I) chloride (25.5 mg, 0.13 mmol) and cesium carbonate (429 mg, 1.31 mmol) in dimethylacetamide (6.5 ml) was degassed with argon for 10 minutes, heated in a sealed tube at 140 C overnight and then cooled to room temperature. Water and ethyl acetate were added, the organic layer was separated, the aqueous layer was extracted twice with ethyl acetate, the combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with 1% HCl/methanol gradient provided compound 31.HCI as white solid. MS (m/z) M+H= 492.1 Synthesis of compound 32:
0 41, o 0F3 Cul, Cs2CO3 10-a + 19-b OH __ lw- NH2 ft 441k N N -' \
I 'N
4101 N N Compound 32 a Scheme 20 A solution of intermediate 10-a (200 mg, 0.55 mmol), intermediate 19-b (225 mg, 0.83 mmol), quinolin-8-ol (16.2 mg, 0.11 mmol), copper (I) iodide (22.0 mg, 0.11 mmol) and cesium carbonate (364 mg, 1.17 mmol) in dimethylacetamide (5.5 ml) was degassed with argon for 10 minutes, heated in a sealed tube at 140 C overnight and then cooled to room temperature.
Water and ethyl acetate were added, the organic layer was separated, the aqueous layer was extracted twice with ethyl acetate. The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with 1% HCl/methanol gradient provided compound 32.HCI as beige solid. MS (m/z) M+H= 546.1 Synthesis of compound 36:
0 41kt u3 cul, Cs2CO3 17-a + 19-b OH __ 11. NH2 0 O
' N N Compound 36 1 / o Scheme 21 A solution of intermediate 17-a (200 mg, 0.53 mmol), intermediate 19-b (215 mg, 0.80 mmol), quinolin-8-ol (15.5 mg, 0.11 mmol), copper (I) iodide (20.3 mg, 0.11 mmol) and cesium carbonate (348 mg, 1.06 mmol) in dimethylacetamide (5.3 ml) was degassed with argon for 10 minutes, heated in a sealed tube at 140 C overnight and then cooled to room temperature.
Water and ethyl acetate were added, the organic layer was separated, the aqueous layer was extracted twice with ethyl acetate. The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Puriflcation by reverse phase chromatography eluting with 1% HCl/methanol gradient provided compound 36=HCI as beige solid. MS (m/z) M+H= 562.2 Synthesis of compound 20:
to 0 Ph3P, DIAD, 0 TBAF 0 HO __________________ ).. i.
OTBS
CF3 las OTBS OH
11-a 22-b HO 22-a 0 . CF3 CuCI, Cs2CO3, 12-a + 22-b ' NH
N N \
i I N
' 101 N N
\ Compound 20 Scheme 22 Step 1: Intermediate 22-a To a solution of 2-(trifluoromethyl)phenylmethanol (1.43 g, 8.10 mmol) in THF (8.10 mL) were sequentially added intermediate 11-a (2.0 g, 8.91 mmol), triphenylphosphine (2.55 g, 9.72 mmol) and DIAD (1.89 ml, 9.72 mmol) drop wise at room temperature. The reaction was then stirred overnight at room temperature. Saturated aqueous ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure.
Purification by silica gel chromatography provided intermediate 22-a as a colorless oil.
Step 2: Intermediate 22-b Tetrabutylammonium fluoride trihydrate (1.81 g, 5.75 mmol) was added to a solution of intermediate 22-a (2.2 g, 5.75 mmol) in THF (23 mL) and the reaction was stirred at room temperature for 1 hour. Saturated aqueous ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 22-b as a colorless oil.
Step 3: Compound 20 A solution of intermediate 12-a (200 mg, 0.65 mmol), intermediate 22-b (309 mg, 1.15 mmol), quinolin-8-ol (19.1 mg, 0.13 mmol), copper (I) chloride (13.0 mg, 0.13 mmol) and cesium carbonate (429 mg, 1.31 mmol), in dimethylacetamide (6.5 ml), was degassed with argon for 10 minutes, heated in a sealed tube at 140 C overnight and then cooled to room temperature. Water and ethyl acetate were added, the organic layer was separated, the aqueous layer was extracted twice with ethyl acetate, the combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% HCl/methanol gradient provided compound 20.1-1CI as beige solid. MS (m/z) M+H= 492.1 Synthesis of compound 29:
0 =
cF3 Cul, Cs2CO3 10-a + 22-b OH __ 1.- NH2 O 4411k N N'' \N
I
0 N N Compound 29 a Scheme 23 A solution of intermediate 10-a (200 mg, 0.55 mmol), intermediate 22-b (225 mg, 0.83 mmol), quinolin-8-ol (16.2 mg, 0.11 mmol), copper (I) iodide (21.2 mg, 0.11 mmol) and cesium carbonate (364 mg, 1.11 mmol), in dimethylacetamide (5.5 ml), was degassed with argon for 10 minutes, heated in a sealed tube at 140 C overnight and then cooled to room temperature. Water and ethyl acetate were added, the organic layer was separated, the aqueous layer was extracted twice with ethyl acetate, the combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% HCl/methanol gradient provided compound 29=HCI as beige solid. MS (m/z) M+H= 546.2 Synthesis of compound 23:
0 =
cF3 Cul, Cs2CO3 17-a + 22-b OH __ ii. NH2 0 40 N N \N
I N
Ol / N _.....) Compound Scheme 24 A solution of intermediate 17-a (200 mg, 0.53 mmol), intermediate 22-b (215 mg, 0.80 mmol), quinolin-8-ol (15.5 mg, 0.11 mmol), copper (I) iodide (20.4 mg, 0.11 mmol) and cesium carbonate (348 mg, 1.07 mmol), in dimethylacetamide (5.3 ml), was degassed with argon for 10 minutes, heated in a sealed tube at 140 C overnight and then cooled to room temperature. Water and ethyl acetate were added, the organic layer was separated, the aqueous layer was extracted twice with ethyl acetate, the combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% HCl/methanol gradient provided compound 23.1-1CI as beige solid. MS (m/z) M+H= 562.1 Synthesis of compound 30:
NC
=
OH
K2CO3 10 =
=
OH Br 4101 OH
CN 25-a CN
Cul, Cs2CO3, 12-a + 25-a ________________ = OH NH2 4Ik \
N
Compound 30 Scheme 25 Step 1: Intermediate 25-a To a solution of 2-(bromomethypbenzonitrile (1.0 g, 5.10 mmol) and resorcinol (2.81 g, 25.5 mmol) in acetone (51.0 mL) was added cesium carbonate (3.32 g, 10.20 mmol) and the reaction was then stirred at reflux for 2 hours. Volatiles were removed under reduced pressure. Saturated aqueous ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 25-a as white solid.
Step 2: Compound 30 A solution of intermediate 12-a (200 mg, 0.65 mmol), intermediate 25-a (222 mg, 0.98 mmol), quinolin-8-ol (19.1 mg, 0.13 mmol), copper (I) iodide (25.0 mg, 0.13 mmol) and cesium carbonate (429 mg, 1.31 mmol), in dimethylacetamide (6.5 ml), was degassed with argon for 10 minutes, heated in a sealed tube at 140 C overnight and then cooled to room temperature. Water and ethyl acetate were added, the organic layer was separated, the aqueous layer was extracted twice with ethyl acetate, the combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% HCl/methanol gradient provided compound 30=HCI as beige solid. MS (m/z) M+H= 449.4 Synthesis of compound 12:
CuCI, Cs2CO3 0 10-a + 25-a ___________________ s NH2 fa O
OH
N N
I 1' .1 .' N N - Compound 12 a Scheme 26 A solution of intermediate 10-a (200 mg, 0.55 mmol), intermediate 25-a (220 mg, 0.98 mmol), quinolin-8-ol (16.2 mg, 0.11 mmol), copper (I) chloride (11.0 mg, 0.11 mmol) and cesium carbonate (546 mg, 1.67 mmol), in dimethylacetamide (5.5 ml), was degassed with argon for 10 minutes, heated in a sealed tube at 140 C overnight and then cooled to room temperature. Water and ethyl acetate were added, the organic layer was separated, the aqueous layer was extracted twice with ethyl acetate, the combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% HCl/methanol gradient provided compound 12=HCI as a beige solid. MS (m/z) M+H= 503.2 Synthesis of compound 35:
0 . CN
Cul, Cs2CO3 0 17-a + 25-a __________________ D.- NH2 . 0 OH
N I N.' \N ' 11101 .7N
N ¨ Compound 35 o Scheme 27 A solution of intermediate 17-a (200 mg, 0.55 mmol), intermediate 25-a (181.0 mg, 0.80 mmol), quinolin-8-ol (15.5 mg, 0.11 mmol), copper (I) iodide (20.3 mg, 0.11 mmol) and cesium carbonate (348 mg, 1.07 mmol), in dimethylacetamide (5.3 ml), was degassed with argon for 10 minutes, heated in a sealed tube at 140 C overnight and then cooled to room temperature. Water and ethyl acetate were added, the organic layer was separated, the aqueous layer was extracted twice with ethyl acetate, the combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse CA 02813299 2013-04-17 .
phase chromatography eluting with a 1% HCl/methanol gradient provided compound 35=HCI as beige solid. MS (m/z) M+H= 519.2 Synthesis of compound 10:
OMe OMe 0 . OTBS Ph3P, DIAD, 110 0 40 TBAF I* 0 0 _____________________ 3.
OTBS OH
OH HO 1.1 OMe 28-b 11-a 28-a O, OMe I, 410 10-a + 28-b CuCCs2CO3, 1.. NH2 O
OH
N INV \N
I ' aCompound 10 Scheme 28 Step 1: Intermediate 28-a To a solution of (3-methoxyphenyl)methanol (1.38 g, 10.0 mmol) in THF
(20.0 mL) were sequentially added intermediate 11-a (2.69 g, 12.0 mmol), triphenylphosphine (3.15 g, 12.0 mmol) and DIAD (2.36 ml, 12.0 mmol) drop wise at room temperature and the reaction was then stirred overnight at room temperature. A saturated aqueous solution of ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure.
Purification by silica gel chromatography provided intermediate 28-a as a colorless oil.
Step 2: Intermediate 28-b Tetrabutylammonium fluoride trihydrate (2.88 g, 9.14 mmol) was added to a solution of intermediate 28-a (2.1 g, 6.10 mmol) in THF (10 mL) and the reaction was stirred at room temperature overnight. A saturated aqueous solution of ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 28-b as a colorless oil.
Step 3: Compound 10 A solution of intermediate 10-a (200 mg, 0.65 mmol), intermediate 28-b (225 mg, 0.97 mmol), quinolin-8-ol (16.2 mg, 0.11 mmol), copper (I) chloride (11.0 mg, 0.1 mmol) and cesium carbonate (546 mg, 1.67 mmol), in dimethylacetamide (5.5 ml), was degassed with argon for 10 minutes, heated in a sealed tube at 140 C overnight and then cooled to room temperature. Water and ethyl acetate were added, the organic layer was separated, the aqueous layer was extracted twice with ethyl acetate, the combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% HCl/methanol gradient provided compound 10.1-1C1 as a yellow solid. MS (m/z) M+H= 508.1 Synthesis of intermediate 29-i Et01=184. H2N)L= , N
, HC(0)0Et 0 toluene, reflux 0 29-a 29-c 29-b 29-c LiAIH4 Ho, j--\\_ 29-d F OMe BBr3 F OH Base F OH
TBDMSCI
=
OMe OH OTBDMS
29-e 29-f 29-g F =
TBAF
29-g Ph3P, DIAD F
HOJ , N
OTBDMS
S¨ OH
29-d 29-h 29-i Scheme 29 Step 1: Intermediate 29-b Ethyl chloroacetate, 29-a (50.0 g, 0.41 mol), and ethyl formate (30.2 g, 0.41 mol) were taken in anhydrous toluene (500 mL) and cooled to 0 C. Sodium ethoxide (35.1 g, 0.49 mol) was added portion wise. The reaction mixture was stirred at 0 C for 5 hours and then at room temperature overnight. The reaction mixture was quenched with water (250 mL) and washed twice with diethyl ether. The aqueous layer was cooled to 0 C and acidified to pH 4-5 using 1 N HCI. The aqueous layer was extracted twice with diethyl ether; the combined organic layers were dried over MgSO4 filtered and concentrated under reduced pressure to provide intermediate 29-b as beige Oil.
Step 2: Intermediate 29-c To a solution of ethyl 2-chloro-3-oxopropanoate, 29-b (34.7 g, 230 mmol), in toluene (250 ml) was added thioacetamide (26.0 g, 346.0 mmol), the reaction was stirred at 90 C overnight and then cooled to room temperature, diluted with water (300 mL) and then neutralized to pH 7 with a saturated aqueous solution of NaHCO3. Ethyl acetate was added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 29-c as beige oil.
Step 4: Intermediate 29-d To a solution of intermediate 29-c (22.2 g, 130.0 mmol) in THF (430 ml) cooled to 0 C was added a 1.0 M solution of LiAIH4in THF (91.0 ml, 91.0 mmol) and the solution was slowly warmed to room temperature and stirred for 2 hours. Water (3.5 ml) was slowly added, followed by 3.5 ml 15% NaOH
(3.5 ml) and water (10.5 ml) and the mixture was stirred for 1 hour. The reaction was filtered over celite and volatiles were removed in vacuo to provide intermediate 29-d as yellow oil.
Step 5: Intermediate 29-f To a solution of 1-fluoro-3,5-dimethoxybenzene (12.5 g, 80 mmol) in dichloromethane (80 ml), cooled to OK, was added 1.0 M solution of boron tribromide in dichloromethane (200 ml, 200 mmol), drop wise over a period of 30 minutes. The reaction was stirred for 1 hour at 0 C and then slowly warmed to room temperature and stirred for 18 hours. The reaction was cooled to 0 C and quenched by the slow addition of Me0H and water. After stirring at room temperature for 1 hour the mixture was filtered and volatiles were removed in vacuo. Ethyl acetate was added to the residue; a precipitate formed and was collected by filtration to provide intermediate 29-f as an orange solid.
Step 6: Intermediate 29-g To a solution of intermediate 29-f (10.25 g, 80.0 mmol) in DMF (50 ml), cooled to 0 C, was added imidazole (5.99 g, 88.0 mmol) and tert-butylchlorodimethylsilane (13.27 g, 88.0 mmol). The reaction was then stirred at room temperature overnight. A saturated aqueous solution of ammonium chloride and ethyl acetate were added, the organic layer was separated, washed 3 times with a saturated aqueous solution of ammonium chloride and brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 29-g as a yellow oil.
Step 7: Intermediate 29-h To a solution of intermediate 29-g (8.0 g, 33.1 mmol) and intermediate 29-d (4.70 g, 36.4 mmol) in THF (20 ml) were sequentially added triphenylphosphine (12.15 g, 46.3 mmol) and DIAD (9.0 ml, 46.3 mmol) at room temperature and the reaction was then stirred at room temperature overnight. Volatiles were removed under reduced pressure. Purification by silica gel chromatography provided intermediate 29-h as a yellow oil.
Step 8: Intermediate 29-i To a solution of intermediate 29-h (6.0 g, 16.97 mmol) in THF (85 ml) was added a 1.0 M solution of TBAF in THF (16.97 ml, 16.97 mmol) and the reaction was stirred at room temperature for 1 hour. A saturated aqueous solution of ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Diethyl ether was added to the residue; a precipitate formed and was collected by filtration to provide intermediate 29-i as white solid.
Synthesis of intermediate 30-b F = OH Ph3P/DIAD
TBAF
HON N
OTBS OTBS OH
29-g 30-a 30-b Scheme 30 Step 1: Intermediate 30-a To a solution of intermediate 29-g (9.0 g, 37.1 mmol) and 2-(methylpyrimidin-5-yl)methanol (4.61 g, 37.1 mmol) in THF (37 ml) were sequentially added triphenylphosphine (11.69 g, 44.6 mmol) and DIAD (9.39 ml, 48.3 mmol) at room temperature and the reaction was then stirred at room temperature for 4 days. Volatiles were removed under reduced pressure. Purification by silica gel chromatography provided intermediate 30-a as a yellow solid.
Step 2: Intermediate 30-b To a solution of intermediate 30-a (12.5 g, 35.9 mmol) in THF (72 ml) was added a 1.0 M solution of TBAF in THF (35.9 ml, 35.9 mmol) and the reaction was stirred at room temperature for 1 hour. A saturated aqueous solution of ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 30-b as a white solid.
Synthesis of intermediate 31-d LiAIH4 31-a 31-b F = OH Ph3P, DD F =01 TBAF
F tio OTBS % OTBS OH
29-g 31-c 31-d Scheme 31 Step 1: Intermediate 31-b To a solution of methyl 6-methylnicotinate 31-a (20.10 g, 133 mmol) in THF
(90 ml) cooled to 0 C was added drop wise a 1.0 M solution of LiAIH4 in THF
(100 ml, 100 mmol) and the reaction was then stirred at 0 C for 1 hour.
Water (3.8 ml) was slowly added, followed by 15% NaOH (3.5 ml) and water (11.4 ml) and the mixture was stirred at room temperature for 1 hour. The reaction was filtered over celite and volatiles were removed in vacuo to provide intermediate 31-b as a yellow oil.
Step 2: Intermediate 31-c To a solution of intermediate 29-g (13.2 g, 54.5 mmol) and intermediate 31-b (7.38 g, 59.9 mmol) in THF (50 ml) were sequentially added triphenylphosphine (21.43 g, 82.0 mmol) and DIAD (17.10 ml, 87.0 mmol) at room temperature and the reaction was then stirred at room temperature for 1 hour. Volatiles were removed under reduced pressure. Purification by silica gel chromatography provided intermediate 31-c as a colorless oil.
Step 3: Intermediate 31-d To a solution of intermediate 31-c (7.6 g, 21.87 mmol) in THF (44 ml) was added tetrabutylammonium fluoride trihydrate (5.72 g, 21.87 mmol) and the reaction was stirred at room temperature for 1 hour. Saturated aqueous ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 31-d as white solid.
Synthesis of intermediate 32-f O rN\\ NaBH4, CaC,I2 OrN1\\ /OH
\--/
32-a 32-b ¨0 PPTS
0, / 0, ¨N\\ _) \-1 OH
HO
32-b 32-c 32-d F io OH Ph3P, DIAD TBAF
TBSO 40. ____________ - HO 40 OTBS 32-d 0 0 N /
29-g 32-e 0 o 32-f 0 Scheme 32 Step 1: Intermediate 32-b To a solution of dimethyl pyridine-2,5-dicarboxylate (13.0 g, 66.6 mmol) in a mixture of THF (110 mL) and ethanol (110 mL) was added calcium chloride (29.6 g, 266 mmol). After stirring at room temperature for 30 minutes the reaction was cooled to 0 C and sodium borohydride (3.78 g, 100 mmol) was added portion wise. After the addition was completed the reaction was stirred at room temperature overnight. A saturated aqueous solution of ammonium chloride and dichloromethane were added, the organic layer was separated and the aqueous phase was extracted twice with dichloromethane. The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure to provide intermediate 32-b as a yellow solid.
Step 2: Intermediate 32-c To a solution of intermediate 32-b (1.70 g, 10.17 mmol) in dichloromethane (203 mL) was added 3,4-dihydro-2H-pyran (4.28 g, 50.8 mmol) and PPTS
(2.56 g, 10.17 mmol) and the reaction was stirred at room temperature overnight. Water was added and the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure to provide intermediate 32-c as a white solid.
Step 3: Intermediate 32-d To a solution of intermediate 32-c (2.56 g, 10.17 mmol) in THF (51 ml) cooled to 0 C was added drop wise a 1.0 M solution of DIBALH in hexane (23.39 ml, 23.39 mmol) and the reaction was then stirred at 0 C for 1.5 hour and room temperature overnight. Water (1.0 ml) was slowly added, followed 15% NaOH (3.5 ml) and water (2.3 ml) and the mixture was stirred at room temperature for 30 minutes. The reaction was filtered over celite and volatiles were removed under reduced pressure. Purification by silica gel chromatography provided intermediate 32-d as a yellow oil.
Step 4: Intermediate 32-e To a solution of intermediate 29-g (1.57 g, 6.51 mmol) and intermediate 32-d (2.56 g, 7.17 mmol) in THF (7 ml) were sequentially added triphenylphosphine (2.56 g, 9.77 mmol) and DIAD (2.04 ml, 10.42 mmol) at room temperature and the reaction was then stirred at room temperature overnight. Volatiles were removed under reduced pressure. Purification by silica gel chromatography provided intermediate 32-e as a yellow solid.
Step 5: Intermediate 32-f To a solution of intermediate 32-e (2.2 g, 4.91 mmol) in THF (9.8 ml) was added a 1.0 M solution of TBAF in THF (4.91 ml, 4.91 mmol) and the reaction was stirred at room temperature for 1 hour. A saturated aqueous solution of ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 32-f as a white solid.
Synthesis of intermediate 33-a -o riY1 y W
=MCP BA _ F 1Wa& 0,....)-OTBS OH
31-c 33-a Scheme 33 To a solution of intermediate 31-c (424 mg, 1.82 mmol) in dichloromethane (9.0 ml) was added m-CPBA (538 mg, 2.18 mmol) and the reaction was stirred at room temperature for 4 hours. A saturated aqueous solution of NaHCO3 and dichloromethane were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 33-a as a white solid.
Synthesis of intermediate 34-d N N
tBuONa ,,E
.,.,1 F 0 F K2c03 F 0 F id o toluene/DMPU F=
0 0 N HC!
F ir N
1 ___________________________________________________________________ \
MOMCIN
OH OMOM HO r¨, \.... OMOM OH
N
34-a 34-b I 34-c 34-d Scheme 34 Step 1: Intermediate 34-b To a solution of 3,5-difluorophenol (15.0 g, 115 mmol) in acetone (200 ml) was added K2CO3 (23.90 g, 173 mmol) and bromomethyl methyl ether (15.85 g, 127 mmol). The reaction was then stirred at room temperature overnight and filtered. The filtrate was concentrated under reduced pressure to provide intermediate 34-b as a colorless oil. , Step 2: Intermediate 34-c To a solution of (1-methyl-1H-imidazol-5-y1) methanol (3.1 g, 27.6 mmol) and intermediate 34-b (4.01 g, 23.04 mmol) in toluene (25.0 ml) and DMPU
(25.0 ml) was added sodium 2-methylpropan-2-olate (4.43 g, 46.1 mmol).
The reaction was stirred overnight at 80 C and then cooled to room temperature. A saturated aqueous solution of ammonium chloride and ethyl acetate were added, the organic layer was separated, washed twice with a saturated aqueous solution of ammonium chloride and brine, dried over MgSO4, filtered and concentrated in vacuo. Purification by silica gel chromatography provided intermediate 34-c as beige oil.
Step 3: Intermediate 34-d To a solution of intermediate 34-c (3.2 g, 12.02 mmol) in Me0H (25.0 ml) was added 4N HCI in dioxane (10.95 ml, 361.0 mmol) and the reaction was stirred overnight at room temperature. Volatiles were removed in vacuo.
Diethyl ether was added to the residue; a precipitate formed and was collected by filtration to provide intermediate 34-d-FICI as a white solid.
Synthesis of intermediate 35-d LiAIH4 HOJS
35-a 35-b tBua F F
toluONene/DMPU
HCI
, N
OMOM OMOM OH
34-h 35-c 35-d Scheme 35 Step I: Intermediate 35-b To a solution of 1,2-dimethy1-1H-imidazole-5-carbaldehyde (1.0 g, 8.06 mmol) in THF (40.3 mL) cooled to 0 C was added drop wise a 1.0 M solution of LiAlHain THF (6.04 ml, 6.04 mmol) and the reaction was then stirred at room temperature for 1 hour. Water (250 uL) was slowly added, followed by 15% NaOH (250 uL) and water (750 uL) and the mixture was stirred at room temperature for 1 hour. The reaction was filtered over celite and volatiles were removed in vacuo to provide intermediate 35-b as a white solid.
Step 2: Intermediate 35-c To a solution of intermediate 35-b (1.50 g, 11..89 mmol) and intermediate 34-b (2.07 g, 11.89 mmol) in DMPU (11.89 mL) and toluene (11.89 mL) was added sodium 2-methylpropan-2-olate (3.43 g, 35.7 mmol) at room temperature. The reaction was stirred overnight at 80 C and then cooled to room temperature. A saturated aqueous solution of ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure.
Purification by silica gel chromatography provided intermediate 35-c as a yellow oil.
Step 3: Intermediate 35-d To a solution of intermediate 35-c (3.30 g, 11.77 mmol) in Me0H (36.2 mL) was added 4N HCI in dioxane (10.7 mL, 353 mmol) and the reaction was stirred at room temperature overnight. Volatiles were removed under reduced pressure. Diethyl ether was added to the residue; a precipitate formed and was collected by filtration to provide intermediate 35-d=HCI as a white solid.
Synthesis of intermediate 36-f N ,N
n ,'-O--H
__________________________ .athj ,-SH
OH KSCN HON H2W04, H20.
HO,..f -N
0H0 Acetic acid ?
?
36-a 112N---- WI
36-b = 114 36-c 36-d N N
tBuONa o j-toluene/DMPU
__________________________________________________ ..
IW-36-d 34-b 36-e 0 36-f It Scheme 36 Step 1: Intermediate 36-c To a suspension of 2-(benzyloxy)ethanamine HCI, 36-b (2.08 g, 11.10 mmol), and 2,5-bis(hydroxymethyl)-1,4-dioxane-2,5-diol 36-a (2.00 g, 11.10 mmol) in iPrOH (8 mL), were sequentially added potassium thiocyanate (1.62 g, 16.7 mmol) and acetic acid (2.03 mL, 35.5 mmol) drop wise. The mixture was stirred at room temperature overnight. Water was added; a precipitate formed and was collected by filtration to provide intermediate 36-c as a white solid.
Step 2: Intermediate 36-d To a solution of intermediate 36-c (1.5 g, 5.67 mmol) and H2W04 (14 mg, 0.057 mmol) in Me0H (22.7 mL) at 40 C was added H202 (1.85 mL, 18.16 mmol) drop wise. The mixture was stirred at reflux overnight and then cooled to room temperature. Volatiles were removed under reduced pressure. Purification by silica gel chromatography provided intermediate 36-d as a colorless oil.
Step 3: Intermediate 36-e To a solution of intermediate 36-d (1.46 g, 6.32 mmol) and intermediate 34-b (1.0 g, 5.74 mmol) in DMPU (11.48 ml) and toluene (11.48 ml) was added sodium 2-methylpropan-2-olate (1.10 g, 11.48 mmol) at room temperature.
The reaction was stirred overnight at 80 C and then cooled to room temperature. A saturated aqueous solution of ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure.
Purification by silica gel chromatography provided intermediate 36-e as a colorless oil.
Step 4: Intermediate 36-f To a solution of intermediate 36-e (400 mg, 1.03 mmol) in Me0H (10.4 mL) was added 4N HCI in dioxane (2.50 mL, 10.0 mmol) the reaction was stirred at room temperature overnight. Volatiles were removed under reduced pressure to provide intermediate 364.1-1C1 as a white solid.
Synthesis of intermediates 37-f and 37-f' SiMe3 SiMe3 N r r_N r NaBH4 NaH (:).M + N HOõ.N, +
N
Me3S1 ' Oji 0) HO,(Ntl H (3'CI 0 ,) 37-a 37-h 37-b 37-c 37-d SiMe3 SiMe3 TMS
( tBuONa F toluene/DMPU --N N,---- \
F 0 F K2CO3 F io cc:
.
F F =
0-- \
OH Br 0 OBn 37-c + 37-d w- to L-TMS
34-a 37-d 0 37-e 0 37-e' TMS
( (N____\\ N,:-- \
r 37-e + 37e' H2, Pd/C N
----.- +
F i, o 1.1 ---\---TMS
HO 374 HO 37-f Scheme 37 Step 1: Intermediates 37-b and 37-b' To a solution of 1H-imidazole-5-carbaldehyde, 37-a (3.0 g, 31..2 mmol), in DMF (20 ml) was added a 60% dispersion of NaH in mineral oil (1.25 g, 31.2 mmol) portion wise. After stirring for 30 minutes at room temperature, (2-(chloromethoxy)ethyl)trimethylsilane (5.73 g, 34.3 mmol) was added and the reaction was then was stirred at room temperature overnight. A
saturated aqueous solution of ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediates 37-b and 37-b' as an inseparable mixture.
Step 2: Intermediates 37-c and 37-c' To a solution of intermediates 37-b and 37-b' (3.2 g, 14.14 mmol) in THF
(56.6 ml) was added NaBH4 (535 mg, 14.14 mmol) at room temperature.
The reaction was stirred overnight at room temperature and then cooled to 0 C. A saturated aqueous solution of ammonium chloride and ethyl acetate were added, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate. The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure to provide intermediate 37-c and 37-c' as an inseparable mixture.
Step 3: Intermediate 37-d To a solution of 3,5-difluorophenol (8.0 g, 61.4 mmol) in acetone (100 ml) was added potassium carbonate (17.0 g, 123.0 mmol) and potassium iodide (1.021 g, 6.15 mmol). The reaction was heated to 65 C and benzyl bromide (8.03 g, 67.6 mmol) was added. The reaction was then stirred overnight at 65 C, cooled to room temperature and filtered. Volatiles were removed in vacuo. A saturated aqueous solution of ammonium chloride and ethyl acetate were added to the residue, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated in vacuo. Purification by silica gel chromatography provided intermediate 37-d as a colorless oil.
Step 4: Intermediate 37-e and 37-e' To a solution of intermediates 37-c and 37-c' (1.0 g, 4.38 mmol) and intermediate 37-d (877 mg, 3.98 mmol) in DMPU (7.96 ml) and toluene (7.96 ml) was added sodium 2-methylpropan-2-olate (765 mg, 3.98 mmol) at room temperature. The reaction was stirred overnight at 80 C and then cooled to room temperature. A saturated aqueous solution of ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediates 37-e and 37-e' as an inseparable mixture.
Step 5: Intermediates 37-f and 37-f' A methanol solution of intermediate 37-e and 37-e' (140 mg, 0.32 mmol) was treated with 10% palladium on carbon (70 mg, 0.045 mmol) and purged with H2. The solution was stirred under H2 (1 atm) for 2 hours before being filtered through celite. The filtrate was concentrated in vacuo to provide intermediate 37-f and 37-f' as an inseparable mixture.
Synthesis of intermediates 38-e and 38-e' Me3Si Me3Si N
i O HO,Z¨N¨
N
isi'---N NaBH4 NaH H o) + )+ <o ---- ___________________________________________ a OyfN ' H
H 38-h Me3Si,...,,O.,.C1 N--( N--( 4 38-h 38-c 38-d 'l SiMe3 SiMe3 y 38-a "H NO
TMS
(o tBuOK ( N--7----F 0 F toluene/DMPU N--(' F
r)N 0 _____________________ . +
F
0 0 --\-_ 38-c + 38-c' TMS
tw- 0 37-d 00:1 0 38-d 0 38-d' 40 I.
TMS
(o (N4 N=---H2, Pd/C rk,N r ,./../N---\0 38-d + 38-d _______ . +
F00 F 40 0 --\__.
TMS
OH 38-e HO 38-e' Scheme 38 Step 1: Intermediates 38-b and 38-b' To a solution of 1H-imidazole-5-carbaldehyde (5.0 g, 45.4 mmol) in DMF (20 mL) was added a 60% dispersion of NaH mineral oil (1.81 g, 45.4 mmol) portion wise. After stirring for 30 minutes at room temperature, (2-(chloromethoxy)ethyl)trimethylsilane (9.08 g, 54.5 mmol) was added and the reaction was then stirred at room temperature overnight. A saturated aqueous solution of ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediates 38-b and 38-h' as an inseparable mixture.
Step 2: Intermediates 38-c and 38-c' To a solution of intermediate 38-b and 38-b' (7.0 g, 29.1 mmol) in THF
(116.0 ml) was added NaBH4 (1.10 g, 29.1 mmol) at room temperature. The reaction was stirred overnight at room temperature and then cooled to 0 C.
A saturated aqueous solution of ammonium chloride and ethyl acetate were added, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate. The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure to provide intermediates 38-c and 38-c' as an inseparable mixture.
Step 3: Intermediate 38-d and 38-d' To a solution of intermediate 38-c and 38-c' (1.0 g, 4.13 mmol) and intermediate 37-d (826 mg, 3.75 mmol) in DMPU (7.50 ml) and toluene (7.50 ml) was added sodium 2-methylpropan-2-olate (721 mg, 7.50 mmol) at room temperature. The reaction was stirred overnight at 80 C and then cooled to room temperature. A saturated aqueous solution of ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediates 38-d and 38-d' as an inseparable mixture.
Step 4: Intermediates 38-e and 38-e' A methanol solution of intermediates 38-d and 38-d' (200 mg, 0.45 mmol) was treated with 10% palladium on carbon (96 mg, 0.045 mmol) and purged with H2. The solution was stirred under H2 (1 atm) for 2 hours before being filtered through celite. The filtrate was concentrated in vacuo to provide intermediate 38-e and 38-e' as an inseparable mixture.
Synthesis of intermediate 39-b F = OH Ph3P, DD F = F
TBAF
HON
OTBS \ c \ OTBS OH
29-g 39-a 39-b Scheme 39 Step 1: Intermediate 39-a To a solution of intermediate 29-g (4.20 g, 17.3 mmol) and pyrimidin-5-ylmethanol (1.90 g, 17.3 mmol) in THF (35 mL) were sequentially added triphenylphosphine (5.91 g, 22.5 mmol) and DIAD (4.38 mL, 22.5 mmol) at room temperature and the reaction was then stirred at room temperature for 3 hours. Volatiles were removed under reduced pressure. Purification by silica gel chromatography provided intermediate 39-a as a white solid.
Step 2: Intermediate 39-b To a solution of intermediate 39-a (5.80 g, 17.3 mmol) in THF (35 mL) was added a 1.0 M solution of TBAF in THF (17.3 ml, 17.3 mmol) and the reaction was stirred at room temperature for 1 hour. A saturated aqueous solution of ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 39-b as a white solid.
Synthesis of intermediate 40-b rN,n F OH Ph3P, DIAD F = 0,,N) TBAF F =
ON) HO
OTBS \ OTBS OH
29-g 40-a 40-b Scheme 40 Step 1: Intermediate 40-a To a solution of intermediate 29-g (4.62 g, 19.1 mmol) and pyrazin-2-ylmethanol (2.10 g, 19.1 mmol) in THF (38 mL) were sequentially added triphenylphosphine (7.50 g, 28.6 mmol) and DIAD (5.19 ml, 26.7 mmol) at room temperature and the reaction was then stirred at room temperature overnight. Volatiles were removed under reduced pressure. Purification by silica gel chromatography provided intermediate 40-a as a colorless oil.
Step 2: Intermediate 40-b To a solution of intermediate 40-a (3.40 g, 10.2 mmol) in THF (20 mL) was added a 1.0 M solution of TBAF in THF (10.2 ml, 10.2 mmol) and the reaction was stirred at room temperature for 1 hour. A saturated aqueous solution of ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 40-b as a white solid.
Synthesis of intermediate 41-b F = OH Ph3P, DIAD F io 0 lel TBAF F = 0 I.
HO
OTBS OTBS OH
29-g 41-a 41-b Scheme 41 Step 1: Intermediate 41-a To a solution of intermediate 29-g (2.60 g, 10.7 mmol) and benzyl alcohol (1.39 g, 12.9 mmol) in THF (20 mL) were sequentially added triphenylphosphine (3.94 g, 15.02 mmol) and DIAD (2.92 mL, 15.0 mmol) at room temperature and the reaction was then stirred at room temperature overnight. Volatiles were removed under reduced pressure. Purification by silica gel chromatography provided intermediate 41-a as colorless oil.
Step 2: Intermediate 41-b To a solution of intermediate 41-a (1.40 g, 4.21 mmol) in THF (10 ml) was added a 1.0 M solution of TBAF in THF (4.63 ml, 4.63 mmol) and the reaction was stirred at room temperature overnight. A saturated aqueous solution of ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 41-b as a colorless oil.
Synthesis of intermediate 42-d Boc NNH NaBH3CN HN,NH2 0=0 ______________________ HCI HCI
,N
H2N , Boc 42-a 42-b 42-c Br Br TEA
NC NC
N
CN 42-c 9-b 42-d Scheme 42 Step 1: Intermediate 42-b To a solution of cyclopentanone (10.00 g, 119.0 mmol) in Me0H (594 mL), were added tert-butyl hydrazinecarboxylate (16.50 g, 125.0 mmol) and the reaction was stirred overnight at room temperature. Volatiles were removed under reduced pressure to provide intermediate 42-b as a white solid.
Step 2: Intermediate 42-c To a solution of intermediate 42-b (10.00 g, 50.40 mmol) in THF (50.4 mL) and Me0H (50.4 mL) was added sodium cyanoborohydride (3.80 g, 60.5 mmol) portion wise. The reaction was refluxed under argon for 10 minutes, and then cooled to room temperature. 6N NCI (25 mL) was added, the mixture was refluxed for 3 hours, cooled to room temperature and stirred overnight. The reaction was filtered to remove inorganic insoluble material and the filtrate was concentrated under reduced pressure and azeotroped three times with toluene. The residue was dissolved in hot isopropanol, cooled to room temperature, diluted with ether and then cooled to 0 C. A
precipitate formed and was collected by filtration to provide intermediate 42-c=FICI as a white solid.
Step 3: Intermediate 42-d To a solution of intermediate 9-b (3.00 g, 11.4 mmol) and TEA (3.50 mL, 25.1 mmol) in Et0H (11.4 mL) was added intermediate 42-c=HCI (1.86 g, 13.7 mmol) and the reaction was then stirred for 2 hours at 100 C. Volatiles were removed under reduced pressure. A saturated aqueous solution of ammonium chloride and ethyl acetate were added to the residue, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 42-d as a white solid.
Synthesis of intermediate 43-e Boc Boc NI' NH
HN
H Pd/CNH HCI HNNH2 0\ 2 . _______________ . HCI
N, H2N Boc 43-a 43-b 43-c 43-d Br Br TEA
NC NC
N
CN 43-d 9-b 43-e Scheme 43 Step 1: Intermediate 43-b To a solution of dihydro-2H-pyran-4-(3H)-one (15.0 g, 150.0 mmol) in Me0H
(749 mL), were added tert-butyl hydrazinecarboxylate (20.79 g, 157.0 mmol) and the reaction was stirred overnight at room temperature. Volatiles were removed under reduced pressure to provide intermediate 43-b as a white solid.
Step 2: Intermediate 43-c A methanol solution of intermediate 43-b (32.1 g, 150.0 mmol) was treated with 10% palladium on carbon (6.39 g, 3.00 mmol), acetic acid (100 pL) and purged with H2. The solution was stirred under H2 (1 atm) overnight before being filtered through celite. The filtrate was concentrated in vacuo to provide intermediate 43-c as a white solid.
Step 3: Intermediate 43-d To a solution of intermediate 43-c (32.4 g, 150 mmol) in Me0H (300 mL) was added 4N HCI in 1,4-dioxane (300 ml, 1200 mmol) and the reaction was stirred at room temperature for 5 hours. Diethyl ether was added and a precipitate formed which was collected by filtration to provide intermediate 43-d=HCI as a white solid.
Step 4: Intermediate 43-e To a solution of intermediate 9-b (5.00 g, 19.0 mmol) and TEA (5.30 mL, 38.0 mmol) in Et0H (19.0 mL) was added intermediate 43-c=FICI (3.48 g, 22.81 mmol) and the reaction was then stirred for 2 hours at 100 C.
Volatiles were removed under reduced pressure. A saturated aqueous solution of ammonium chloride and ethyl acetate were added to the residue, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure to provide intermediate 43-e as a yellow solid.
Synthesis of intermediate 44-d Boc NNH
H _________________________ A. I
V\ NaBH3CN
HCI HN,NH2 _________________________________________________ p riN
N, H2N, Boc 44-a 44-b 44-c Br Br SI ilk TEA
NC ________________________________________ A NC
0 I \ N
44-c CN
H2N "....,,_ 9-b 44-d Scheme 44 Step 1: Intermediate 44-b tert-Butyl hydrazinecarboxylate (7.60 g, 57.5 mmol) was added to acetone (50 mL) and the reaction was stirred overnight at room temperature.
Volatiles were removed under reduced pressure to provide intermediate 44-b as a white solid.
Step 2: Intermediate 44-c To a solution of intermediate 44-b (9.90 g, 57.5 mmol) in THF (57.5 mL) and Me0H (57.5 mL) was added sodium cyanoborohydride (4.34 g, 69.0 mmol) portion wise. The reaction was refluxed under nitrogen for 10 minutes, and then cooled to room temperature. 6N HCI (30 mL) was added, the mixture was refluxed for 3 hours, cooled to room temperature and stirred overnight.
The reaction was filtered to remove inorganic insoluble material and the filtrate was concentrated under reduced pressure and azeotroped three times with toluene for complete water removal. The residue was dissolved in hot isopropanol, cooled to room temperature, diluted with ether and then cooled to 0 C. A precipitate formed and was collected by filtration to provide intermediate 44-c=HCI as a white solid.
Step 3: Intermediate 44-d To a solution of intermediate 9-b (12.61 g, 47.9 mmol) and TEA (14.70 mL, 105.0 mmol) in Et0H (96.0 ml) was added intermediate 44-c.HCI (6.36 g, 57.5 mmol) and the reaction was then stirred for 2 hours at 100 C. Volatiles were removed under reduced pressure. A saturated aqueous solution of ammonium chloride and ethyl acetate were added to the residue, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 44-d as a white solid.
Synthesis of intermediate 45-a Br Br TEA
NC __________________________________________________ . NC410 / 0 \
I N
CN) NHNH2 N
/)----9-b 45-a Scheme 45 To a solution of intermediate 9-b (2.0 g, 7.60 mmol) and TEA (2.12 ml, 15.2 mmol) in Et0H (7.60 mL) was added tert-butylhydrazine hydrochloride (1.13 g, 9.12 mmol) and the reaction was then stirred for 2 hours at 100 C.
Volatiles were removed under reduced pressure. A saturated aqueous solution of ammonium chloride and ethyl acetate were added to the residue, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure to provide intermediate 45-a as a yellow solid.
Synthesis of intermediate 46-a Br Br TEA
NC NC
0 \ N
9-b 46-a Scheme 46 To a solution of intermediate 9-b (1.45 g, 5.53 mmol) and TEA (1.54 mL, 11.1 mmol) in Et0H (15.0 mL) was added cyclohexylhydrazine hydrochloride (1.00 g, 6.64 mmol) and the reaction was then stirred for 2 hours at 100 C.
Volatiles were removed under reduced pressure. A saturated aqueous solution of ammonium chloride and ethyl acetate were added to the residue, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure to provide intermediate 46-a as a yellow solid.
Synthesis of intermediate 47-a Br Br TEA
NC NC
0 I \ N
CN HO H2 H2N Nv_ 9-b 47-a OH
Scheme 47 To a solution of intermediate 9-b (2.00 g, 7.60 mmol) and TEA (1.27 mL, 9.12 mmol) in Et0H (7.60 mL) was added 2-hydroxyethylhydrazine (618 pL, 9.12 mmol) and the reaction was then stirred for 2 hours at 100 C. Volatiles were removed under reduced pressure. A saturated aqueous solution of ammonium chloride and ethyl acetate were added to the residue, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure to provide intermediate 47-a as a white solid.
Synthesis of intermediate 48-c cK)OH NaOH
Hydrazine 48-a 48-b Br Br TEA
NC NC
N
48-b 9-b 48-c OH
Scheme 48 Step 1: Intermediate 48-b To a mixture of sodium hydroxide (7.37 g, 184.0 mmol) and hydrazine monohydrate (46.10 g, 921.0 mmol) heated to 95 C, was added 1-chloro-2-methylpropan-2-ol (20.00 g, 184.0 mmol). The reaction was stirred overnight at 95 C and then cooled to room temperature. Volatiles were removed under reduced pressure. THF (40 mL) and diethyl ether (40 mL) were added to the residue; a precipitate formed which was removed by filtration. The filtrate was concentrated under reduced pressure to provide intermediate 48-b as a colorless oil.
Step 2: Intermediate 48-c To a solution of intermediate 9-b (4.45 g, 16.9 mmol) and TEA (4.71 mL, 33.8 mmol) in Et0H (15.0 mL) was added intermediate 48-b (1.76 g, 16.9 mmol) and the reaction was then stirred for 2 hours at 100 C. Volatiles were removed under reduced pressure. A saturated aqueous solution of ammonium chloride and ethyl acetate were added to the residue, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure to provide intermediate 48-c as a white solid.
Synthesis of intermediate 49-c NaOH
NOH
Hydrazine 49-a 49-b Br TEA Br NC N
N
49-h CN N
9-b 49-c OH
Scheme 49 Step 1: Intermediate 49-b To a mixture of sodium hydroxide (1.15 g, 28.8 mmol) and hydrazine monohydrate (7.20 g, 144.0 mmol) heated to 95 C, was added 3-bromo-1-propanol (4.00 g, 28.8 mmol) and the reaction was stirred overnight at 95 C
and then cooled to room temperature. Volatiles were removed under reduced pressure. Ethanol was added to the residue; a precipitate formed and was removed by filtration. The filtrate was concentrated under reduced pressure and 1M HCI in diethyl was added to the residue. After stirring for 15 minutes a precipitate formed and was collected by filtration to provide intermediate 49-b=HCI as a white solid.
Step 2: Intermediate 49-c To a solution of intermediate 9-b (1.12 g, 4.28 mmol) and TEA (1.19 mL, 8.56 mmol) in Et0H (10.0 mL) was added intermediate 49-b=FICI (650 mg, 5.13 mmol) and the reaction was then stirred for 2 hours at 100 C. Volatiles were removed under reduced pressure. A saturated aqueous solution of ammonium chloride and ethyl acetate were added to the residue, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure to provide intermediate 49-c as a white solid.
Synthesis of intermediate 50-c Boc n OH FIN,N
NaBH3CN H2N,INH HCI
_____________________________ _ ____________________ .
H
HCI
r-OH N
H2NõBoc OH OH OH OH
36-a 50-a 50-b Br Br TEA
___________________________ .
CN 50-b N
9-b OH 50-c Scheme 50 Step 1: Intermediate 50-a Dihydroxyacetone dimer (15.0 g) and tert-butyl hydrazinecarboxylate (22.01 g) were dissolved in ethanol (500 mL) and this solution was stirred at room temperature for 2 days. After the reaction mixture was concentrated under reduced pressure, the resulting residue was recrystallized from ethyl acetate to provide intermediate 50-a as a white solid.
Step 2: Intermediate 50-b To a solution of intermediate 50-a (10.0 g, 49.0 mmol) in THF (49.0 mL) and Me0H (49.0 mL) was added sodium cyanoborohydride (3.69 g, 58.8 mmol) portion wise. The reaction was refluxed under nitrogen for 10 minutes, and then cooled to room temperature. 6N HCI (40 mL) was added, the mixture was refluxed for 3 hours, cooled to room temperature and stirred overnight.
The reaction was filtered to remove inorganic insoluble material and the filtrate was concentrated under reduced pressure and azeotroped three times with toluene to provide intermediate 50-b=HCI as a white solid.
Step 3: Intermediate 50-c To a solution of intermediate 9-b (10.70 g, 40.9 mmol) and TEA (12.5 mL, 90.0 mmol) in Et0H (40.9 mL) was added intermediate 50-13.1-1C1 (7.00 g, 49.1 mmol) and the reaction was then stirred for 2 hours at 100 C. Volatiles were removed under reduced pressure. A saturated aqueous solution of ammonium chloride and ethyl acetate were added to the residue, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 50-c as a beige solid.
Synthesis of intermediate 51-a Br Br O fa Ph3P, DIAD
NC ________________________________ v NC
l \ N l \ N
H2N [\il HC:Y>C0 H2N
LC) 9-c 51-a Scheme 51 To a solution of intermediate 9-c (1.40 g, 5.32 mmol) and 3-methyloxetan-3-yl)methanol (1.08 g, 10.64 mmol) in THF (5.3 mL) were sequentially added triphenylphosphine (1.67 g, 6.39 mmol) and DIAD (1.13 mL, 5.85 mmol) at room temperature and the reaction was then stirred at room temperature for 4 days. Volatiles were removed under reduced pressure. Purification by silica gel chromatography provided intermediate 51-a as a white solid.
Synthesis of intermediate 52-a NC Br Br Ph3P, DIAD NC
H2N /\N H
1\1 rN N,N
0) 9-c 52-a 170 Scheme 52 To a solution of intermediate 9-c (500 mg, 1.90 mmol) and 3-morpholinopropan-1-ol (263 pl, 1.90 mmol) in THF (19.0 ml) cooled to 0 C
were sequentially added triphenylphosphine (498 mg, 1.90 mmol) and DIAD
(370 pl, 1.90 mmol). The reaction was stirred at 0 C for 1 hour and room temperature overnight. Volatiles were removed under reduced pressure.
Purification by silica gel chromatography provided intermediate 52-a as a white foam.
Synthesis of intermediate 53-a Br Br NH2 = NH2 N Ph3P, DIAD
,N N
N N N iN\
r-N\ 10 9-d 53-a Scheme 53 To a solution of intermediate 9-d (3.00 g, 10.3 mmol) and N-(2-hydroxyethyl)morpholine (1.88 ml, 15.1 mmol) in THF (103 ml) cooled to 0 C were sequentially added triphenylphosphine (4.07 mg, 15.1 mmol) and DIAD (3.02 ml, 15.5 mmol). The reaction was stirred at 50 C overnight.
Volatiles were removed under reduced pressure. Purification by silica gel chromatography provided intermediate 53-a as a white solid.
Synthesis of intermediate 54-a Br Br = 110+
Ph3P, DIAD NC
NC
\ /
9-c 54-a Scheme 54 To a solution of intermediate 9-c (500 mg, 1.90 mmol) and 2-(pyrrolidin-1-yl)ethanol (219 mg, 1.90 mmol) in THF (9.5 ml) cooled to 0 C were sequentially added triphenylphosphine (498 mg, 1.90 mmol) and DIAD (370 pl, 1.90 mmol). The reaction was stirred at 0 C for 1 hour and room temperature for 30 minutes. Volatiles were removed under reduced pressure.
Purification by silica gel chromatography provided intermediate 54-a as yellow solid.
Synthesis of Compound 65:
Br 0 0 it H2N N)-(OH 0 Cul, Cs2CO3 NH2 I' N 31-d N N
bo 17-a Compound 65 Scheme 55 To a solution of intermediate 17-a (321 mg, 0.85 mmol) and intermediate 31-d (200 mg, 0.85 mmol) in 1,4-dioxane (4.30 ml) were sequentially added N,N-dimethylglycine (265 mg, 2.57 mmol), copper(I) iodide (163 mg, 0.85 mmol) and cesium carbonate (1.12 g, 3.43 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate. The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 65.2HCI as a white solid.
Synthesis of Compound 85 Br 0 0 411t NC * Cul, Cs2CO3 tN
HC(OMe)3 =
/ \ N
_______________________________________________ =
/
H2N m 31-d NC N
N
' )\ H2N N
44-d 56-a Compound Scheme 56 Step 1: Intermediate 56-a To a solution of intermediate 44-d (5.00 g, 16.4 mmol) and intermediate 31-d (4.20 g, 18.0 mmol) in 1,4-dioxane (54.6 ml) were sequentially added N,N-dimethylglycine (3.80 g, 36.9 mmol), copper(I) iodide (2.34 g, 12.29 mmol) and cesium carbonate (21.35 g, 65.5 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate.
The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided intermediate 56-a.HCI as a white solid.
Step 2: Compound 85 Intermediate 56-a=FICI (3.13 g, 6.84 mmol) and trimethyl orthoformate (48.7 ml, 445.0 mmol) were heated at 110 C for 3 hours. Excess trimethyl orthoformate was removed in vacuo and the residue was treated with ammonia (7.0 N in Me0H) (48.9 ml, 342.0 mmol). The mixture was stirred at room temperature for 3 days and volatiles were removed under reduced pressure. Purification by reverse phase chromatography eluting with a 1%
aqueous HCl/methanol gradient provided compound 85.2HCI as a white solid.
MS (m/z) M+H= 485.2 Synthesis of Compound 91:
Br Oil 0 0 0 =
NC Cul, Cs2CO3 HC(OMe)3 40 _______________________________________________ =
H \ N \
¨2..N N- 33-a NC N NH3 N
\
) 43-e 57-a 0 Compound 91 Scheme 57 Step 1: Intermediate 57-a To a solution of intermediate 43-e (192 mg, 0.55 mmol) and intermediate 33-a (190 mg, 0.66 mmol) in 1,4-dioxane (2.8 ml) were sequentially added N,N-dimethylglycine (129 mg, 1.24 mmol), copper(I) iodide (79 mg, 0.42 mmol) and cesium carbonate (722 mg, 2.21 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate.
The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided intermediate 57-a as a yellow foam.
Step 2: Compound 91 Intermediate 57-a (286 mg, 0.55 mmol) and trimethyl orthoformate (3.94 ml, 36.0 mmol) were heated at 110 C for 1 hour. Excess trimethyl orthoformate was removed in vacuo and the residue was treated with 7.0 N
ammonia in Me0H (3.96 ml, 27.7 mmol). The mixture was stirred at room temperature for 3 days and volatiles were removed under reduced pressure.
Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 91.HCI as white solid. MS (m/z) M+H= 543.1 Synthesis of Compound 101:
Br Cul, Cs2CO3HC(OMe)3 /\ 0 1. 0 NH2' H2N NN NH3 "N
32-f \
N NiN OH
NC NN
43-eH2N 58-a o Compound 101 o Scheme 58 Step 1: Intermediate 58-a To a solution of intermediate 43-e (400 mg, 1.15 mmol) and intermediate 32-f (384 mg, 1.15 mmol) in 1,4-dioxane (2.8 ml) were sequentially added N,N-dimethylglycine (267 mg, 2.59 mmol), copper(I) iodide (165 mg, 0.86 mmol) and cesium carbonate (1.50 g, 4.61 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate. The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure to provide intermediate 58-a as a beige foam.
Step 2: Compound 101 Intermediate 58-a (690 mg, 1.15 mmol) and trimethyl orthoformate (8.18 ml, 74.8 mmol) were heated at 110 C for 4 days. Excess trimethyl orthoformate was removed in vacuo and the residue was treated with 7.0 N
ammonia in Me0H (8.21 ml, 57.5 mmol). The mixture was stirred at room temperature overnight and volatiles were removed under reduced pressure.
Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 101.2HCI as a white solid. MS
(m/z) M+H= 543.1 Synthesis of Compound 128:
F
Br 0 F
0 =
ii, 1\1I)-LOH
NC / Cul, Cs2CO3 0 lei 0 HC(OMe)3 L
m H2N * 0 \ __________ = .,,,-., NH3 / \ N
32-f N-- 1 '` N
N ' 1 \
.) 40 ,--cAH L 1 ,N OH
N\
\ , 44-d N 59-a Compound 128 H2N ?---Scheme 59 Step 1: Intermediate 59-a To a solution of intermediate 44-d (261 mg, 0.85 mmol) and intermediate 32-f (285 mg, 0.85 mmol) in 1,4-dioxane (2.8 ml) were sequentially added N,N-dimethylglycine (198 mg, 1.92 mmol), copper(I) iodide (122 mg, 0.64 mmol) and cesium carbonate (1.11 g, 3.42 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate. The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure to provide Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided intermediate 59-a.HCI as a beige foam.
Step 2: Compound 128 Intermediate 59-a (405 mg, 0.85 mmol) and trimethyl orthoformate (6.08 ml, 55.6 mmol) were heated at 110 C for 1 hour. Excess trimethyl orthoformate was removed in vacuo and the residue was treated with 7.0 N
ammonia in Me0H (6.11 ml, 42.8 mmol). The mixture was stirred at room temperature for 3 days and volatiles were removed under reduced pressure.
Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 128.2HCI as a white solid. MS
(m/z) M+H= 501.1 Synthesis of Compound 78:
F
Br 1 0 F
Nj=LOH 0 NC 11 Cul, Cs2CO3 o Si o HC(OMe)3 OTh---\-_______________________________________________ r \ *
Ly---\N NH3 NI-12 . S7/ N
H2N /N-N 29-i 40 S--c \
1, 1 ,N
NC\ N N N\
, N
44-d H2N ------ 60-a Compound 78 Scheme 60 Step 1: Intermediate 60-a To a solution of intermediate 44-d (300 mg, 0.98 mmol) and intermediate 29-i (259 mg, 1.08 mmol) in 1,4-dioxane (3.9 ml) were sequentially added N,N-dimethylglycine (304 mg, 2.95 mmol), copper(I) iodide (187 mg, 0.98 mmol) and cesium carbonate (961 mg, 2.95 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate.
The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided intermediate 60-a=HCI as a beige foam.
Step 2: Compound 78 Intermediate 60-a=HCI (265 mg, 0.57 mmol) and trimethyl orthoformate (1.87 ml, 17.16 mmol) were heated at 110 C for 1 hour. Excess trimethyl orthoformate was removed in vacuo and the residue was treated with 7.0 N
ammonia in Me0H (5.7 ml, 11.44 mmol). The mixture was stirred at room temperature overnight and volatiles were removed under reduced pressure.
Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 78.2HCI as beige solid. MS (m/z) M+H= 491.2 Synthesis of Compound 58:
F
Br O=
N,)-(OH
NH2 /1 NH2 Os S(/ N
Cul, Cs2CO3 \
' N N)....,.., 29-i N 14)....._, U U
10-a Compound 58 Scheme 61 To a solution of intermediate 10-a (3.96 g, 11.1 mmol) and intermediate 29-i (2.91 g, 12.2 mmol) in 1,4-dioxane (55.3 ml) were sequentially added N,N-dimethylglycine (3.42 g, 33.2 mmol), copper(I) iodide (2.10 g, 11.07 mmol) and cesium carbonate (10.82 g, 33.2 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate. The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 58.2HCI as a beige solid. MS (m/z) M+H= 517.2 Synthesis of Compound 117:
F
F
Br 0 0 .
.Ij=(OH OTh"----\-0 el 0 NC Cul, Cs2CO3 HC(OMe)3 / \k, _____________ 1- 0 NH2 it ____________________________________ s,,,N
H2N N.- s__c NH3 \
29-i N -- \
NC N NI N'IN1 OH \ , N \_.4......
H2N N\.___1( OH
48-c OH 62-a Compound 117 Scheme 62 Step 1: Intermediate 62-a To a solution of intermediate 48-c (200 mg, 0.59 mmol) and intermediate 29-1 (157 mg, 0.65 mmol) in 1,4-dioxane (1.50 ml) were sequentially added N,N-dimethylglycine (92 mg, 0.89 mmol), copper(I) iodide (57 mg, 0.29 mmol) and cesium carbonate (583 mg, 1.79 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate.
The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided intermediate 62-a=HCI as a beige solid.
Step 2: Compound 117 Intermediate 62-a=HCI (322 mg, 0.65 mmol) and trimethyl orthoformate (3.0 ml, 19.57 mmol) were heated at 110 C for 1 hour. Excess trimethyl orthoformate was removed in vacuo and the residue was treated with ammonia (7.0 N in Me0H) (1.85 ml, 13.0 mmol). The mixture was stirred at room temperature overnight and volatiles were removed under reduced pressure. Purification by reverse phase chromatography eluting with a i%
aqueous HCl/methanol gradient provided compound 117=2HCI as a white solid. MS (m/z) M+H= 521.1 Synthesis of Compound 100:
F
F
Br 1 0 0 40 0 o=
46, -N 0OH 0 Y\N
S/, N
NC Cul, Cs2CO3 S---c HC(OMe)3 __________________ r _________________________ r-/ \ \
\
, "
H2N N,"m 29-i NC N , nu I , N,-.
OH OH
50-c 63-a Compound 100 Scheme 63 Step 1: Intermediate 63-a To a solution of intermediate 50-c (1.42 g, 4.21 mmol) and intermediate 29-i (1.10 g, 4.63 mmol) in 1,4-dioxane (16.8 ml) were sequentially added N,N-dimethylglycine (1.30 g, 12.6 mmol), copper(I) iodide (802 mg, 4.21 mmol) and cesium carbonate (4.12 g, 12.63 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate. The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided intermediate 63-a=HCI as a white solid.
Step 2: Compound 100 Intermediate 63-a=HCI (577 mg, 1.16 mmol) and trimethyl orthoformate (3.82 ml, 35.0 mmol) were heated at 110 C for 3 hours. Excess trimethyl orthoformate was removed in vacuo and the residue was treated with 7.0 N
ammonia in Me0H (3.30 ml, 23.32 mmol). The mixture was stirred at room temperature overnight and volatiles were removed under reduced pressure.
Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 100.2HCI as white solid. MS
(m/z) M+H= 523.2 Synthesis of Compound 113:
0 0 4, 0 ,49 s.õN NH2 sõNi NH2 =41k $,//N
Bu Tsa N N N\,11 N \ N IsLN \.N
OH 0, 0 0 Compound 100 ti\(. 64-a Compound 113 Scheme 64 Step 1: Intermediate 64-a To a solution of compound 100 (198.0 mg, 0.38 mmol) in THF cooled to -C was slowly added a 2.5 M solution of n-butyllithium in THF (304 pl, 0.76 mmol). After stirring for 30 minutes, para-toluenesulfonyl chloride (72.0 mg, 0.38 mmol) in THF (2 ml) was added. The reaction was stirred at 60 C
overnight and then cooled to room temperature. Water and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 64-a as beige foam.
Step 2: Compound 113 To a solution of intermediate 64-a (85.0 mg, 0.12 mmol) in THF cooled to -10 C was slowly added a 2.5 M solution of n-butyllithium in THF (110.0 pl, 0.27 mmol). The reaction was stirred at 60 C overnight and then cooled to room temperature. Water and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided compound 113 as pale yellow foam. MS (m/z) M+H= 505.2 Synthesis of Compound 125:
Br 0 o,0 4.
Cul, Cs2C0 NC 3 HC(OMe)3 S,/,1%1 I \ N I \,N NH3 N".
29-i I ,N
oI N NL7C0 51-a 65-a Compound 125 Scheme 65 Step 1: Intermediate 65-a To a solution of intermediate 51-a (300 mg, 0.86 mmol) and intermediate 29-i (262 mg, 0.95 mmol) in 1,4-dioxane (1.50 ml) were sequentially added N,N-dimethylglycine (267 mg, 2.59 mmol), copper(I) iodide (165 mg, 0.86 mmol) and cesium carbonate (845 mg, 2.59 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate.
The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 65-a as a beige foam.
Step 2: Compound 125 Intermediate 65-a (120 mg, 0.23 mmol), trimethyl orthoformate (260 pl, 2.3 mmol) and PTSA (catalytic) were stirred at room temperature for 1 hour.
Excess trimethyl orthoformate was removed in vacuo and the residue was treated with 7.0 N ammonia in Me0H (652 pl, 4.6 mmol). The mixture was stirred at room temperature for 3 days and volatiles were removed under reduced pressure. Purification by silica gel chromatography provided compound 125 as a white solid. MS (m/z) M+H= 533.2 Synthesis of Compound 54:
Br ')'OH
N Cul, Cs2CO3 I
NN
N NI)Th 29-i N )ThN
17-a Compound 54 Scheme 66 To a solution of intermediate 17-a (1.20 g, 3.21 mmol) and intermediate 29-1 (844 mg, 3.53 mmol) in DMF (16.0 ml) were sequentially added N,N-dimethylglycine (992 mg, 9.62 mmol), copper(I) iodide (611 mg, 3.21 mmol) and cesium carbonate (4.18 g, 12.83 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate. The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 54.2HCI as a yellow solid. MS (m/z) M+H= 533.1 Synthesis of Compound 55:
F F
Br I V 0 . o=
411 -'NOH
* 0 0 NC Cul, Cs2CO3 =:---\¨ formamidine NH2 / \ ______________ ' NC Si/14 ----0-H2N NN 29-1 " \
.._ 52-a 67-a Compound 55 Scheme 67 Step 1: Intermediate 67-a To a solution of intermediate 52-a (397.0 mg, 1.00 mmol) and intermediate 29-1 (268 mg, 1.12 mmol) in 1,4-dioxane (5.0 ml) were sequentially added N,N-dimethylglycine (157 mg, 1.53 mmol), copper(I) iodide (97.0 mg, 050 mmol) and cesium carbonate (995 mg, 3.05 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate.
The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure to provide intermediate 67-a as beige foam.
Step 2: Compound 55 Formamide (2.84 ml, 71.3 mmol) was added to intermediate 67-a (559 mg, 1.0 mmol) and the reaction was stirred at 180 C for 2 hours. Water and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure.
Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 55.3HCI as beige solid. MS (m/z) M+H= 576.2 Synthesis of Compound 52:
F
Br OH
NH2 41k 441k ,---"\--N \ N 0 NH2 * SerNI
I' N N\ Cs2CO3, Cul I N I
---1 29-1 N rµl_ N--\ ---1 53-a (...__ ) (N--) Compound 52 LO
Scheme 68 To a solution of intermediate 53-a (1.0 g, 2.48 mmol) and intermediate 29-i (771 mg, 3.22 mmol) in DMAC (12.4 ml) were sequentially added quinolin-8-ol (36.0 mg, 0.24 mmol), copper(I) iodide (47.0 mg, 0.24 mmol) and cesium carbonate (808 mg, 2.48 mmol) and the reaction was heated at 140 C for 2 hours. A saturated aqueous solution of ammonium chloride and ethyl acetate were added, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate. The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 52.3HCI as white solid. MS (m/z) M+H= 562.1 Synthesis of Compound 57:
F F
Br i 0 0* 0=
11 fh NC / H NC l formamide * --'----\
, Cr¨ \ NH2 N ____________________________________________ p.
1 CulCs2CO3 H2N N=N 29-i I \,N S(/
14-- . \
i ,N I
H2N N, '---1 N NI, µ----1 --J
54-a 69-a Compound 57 Scheme 69 Step 1: Intermediate 69-a To a solution of intermediate 54-a (425.0 mg, 1.18 mmol) and intermediate 29-1 (282 mg, 1.18 mmol) in 1,4-dioxane (5.9 ml) were sequentially added N,N-dimethylglycine (182 mg, 1.77 mmol), copper(I) iodide (112 mg, 0.59 mmol) and cesium carbonate (1.15 g, 3.54 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate. The combined organic extracts were washed with brine, dried over MgS0.4, filtered and concentrated under reduced pressure to provide intermediate 69-a as a beige foam.
Step 2: Compound 57 Formamide (5.53 ml, 139.0 mmol) was added to intermediate 69-a (600 mg, 1.15 mmol) and the reaction was stirred at 180 C for 4 hours. Water and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure.
Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 57.3HCI as beige solid. MS (m/z) M+H= 546.2 Synthesis of Compound 102:
Br O 0 0 *
LrN
NC Cul, Cs2CO3 I 1 ts( HC(OMe)3 NH, N
H2N N=N 30-b NC NH3 I ,N
44-d 70-a Compound 102 Scheme 70 Step 1: Intermediate 70-a To a solution of intermediate 44-d (4.00 g, 13.1 mmol) and intermediate 30-b (3.38 g, 14.4 mmol) in 1,4-dioxane (43.7 ml) were sequentially added N,N-dimethylglycine (3.04 g, 29.5 mmol), copper(I) iodide (1.87 g, 9.83 mmol) and cesium carbonate (17.08 g, 52.4 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate.
The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided intermediate 70-a=HCI as beige foam.
Step 2: Compound 102 Intermediate 70-a=HCI (3.80 g, 7.68 mmol) and trimethyl orthoformate (54.6 ml, 499.0 mmol) were heated at 110 C for 1 hour. Excess trimethyl orthoformate was removed in vacuo and the residue was treated with 7.0 N
ammonia in Me0H (54.8 ml, 384.0 mmol). The mixture was stirred at room temperature for 2 days and volatiles were removed under reduced pressure.
Purification by reverse phase chromatography eluting with a 10/0 aqueous HCl/methanol gradient provided compound 102.2HCI as white solid. MS
(m/z) M+H= 486.2 Synthesis of Compound 129:
F F
Br 1 0 0 0 o o =
-N o 2qj-OH L' NC = Cul, Cs2CO3 0 I I
Thµl HC(OMe)3 NH2 ik /--N
___________________ y __...
i\ m N , \ N--:"--c \
H2N Ny¨ 30-b NC ,,, ,' NI-13 is I ,N
N
N N, /-'-. H2N .-- i\---45-a 71-a Compound 129 Scheme 71 Step 1: Intermediate 71-a To a solution of intermediate 45-a (350 mg, 1.10 mmol) and intermediate 30-b (283 mg, 1.20 mmol) in 1,4-dioxane (4.3 ml) were sequentially added N,N-dimethylglycine (1.07 g, 3.29 mmol), copper(I) iodide (209 mg, 1.10 mmol) and cesium carbonate (985 mg, 3.02 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate.
The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided intermediate 71-a=HCI as a beige solid.
Step 2: Compound 129 Intermediate 71-a=HCI (403.0 mg, 0.85 mmol) and trimethyl orthoformate (6.07 ml, 55.5 mmol) were heated at 110 C for 3 hours. Excess trimethyl orthoformate was removed in vacuo and the residue was treated with 7.0 N
ammonia in Me0H (6.07 ml, 42.7 mmol). The mixture was stirred at room temperature overnight and volatiles were removed under reduced pressure.
Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 129.2HCI as beige solid. MS
(m/z) M+H= 500.1 Synthesis of Compound 106:
F F
Br 1 0 0 Si 0 0*
,4 '-)LOH . 0 L'N
NC it cui,c.2.3 tst HC(OMe)3 NH2 _________________________________________________ /
/ N N \ NI-----.- 30-b NC N N NH3 \ N1_ l ,N
INI___, /c H2N
43-e 72-a Compound 106 Scheme 72 Step 1: Intermediate 72-a To a solution of intermediate 43-e (350 mg, 1.00 mmol) and intermediate 30-b (260 mg, 1.10 mmol) in 1,4-dioxane (4.0 ml) were sequentially added N,N-dimethylglycine (312 mg, 3.02 mmol), copper(I) iodide (192 mg, 1.00 mmol) and cesium carbonate (985 mg, 3.02 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate.
The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided intermediate 72-a.HCI as a beige solid.
Step 2: Compound 106 Intermediate 72-a=HCI (263 mg, 0.52 mmol) and trimethyl orthoformate (1.72 ml, 15.8 mmol) were heated at 110 C for 2 hours. Excess trimethyl orthoformate was removed in vacuo and the residue was treated with 7.0 N
ammonia in Me0H (46.2 ml, 324 mmol). The mixture was stirred at room temperature overnight and volatiles were removed under reduced pressure.
Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 106.2HCI as a beige solid. MS
(m/z) M+H= 528.1 Synthesis of Compound 114:
Br 9 0 el 0 0*
N,2cOH 0 trN
NC Cul, Cs2CO3 110 I I
HC(OMe)3 NH2 fk N \
H2N /N-N 30-b NC N
OH A-OH
49-c 73-a Compound 114 Scheme 73 Step 1: Intermediate 73-a To a solution of intermediate 49-c (250 mg, 0.78 mmol) and intermediate 30-b (201 mg, 0.85 mmol) in 1,4-dioxane (3.1 ml) were sequentially added N,N-dimethylglycine (241 mg, 2.33 mmol), copper(I) iodide (148 mg, 0.78 mmol) and cesium carbonate (761 mg, 2.33 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate.
The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided intermediate 73-a=FICI as a beige solid.
Step 2: Compound 114 Intermediate 73-a=FICI (106 mg, 0.22 mmol) and trimethyl orthoformate (737 pl, 6.74 mmol) were heated at 110 C for 3 hours. Excess trimethyl orthoformate was removed in vacuo and the residue was treated with 7.0 N
ammonia in Me0H (2.24 ml, 4.49 mmol). The mixture was stirred at room temperature overnight and volatiles were removed under reduced pressure.
Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 114.2HCI as a beige solid. MS
(m/z) M+H= 502.2 Synthesis of Compound 130:
F
F
Br 1 0 0 lei 0 0 =
Nji,.OH0 \
H2N Irl NC 11 Cul, Cs2CO3 f*1 HC(OMe)3 NH240 N---;"-k 'N NC N
\ OH \ 30-b NH3 Isr" \
' N
, N I ,N
rJ0H H2N Z-----/
OH ( OH OH
50-c 74-a Compound 130 Scheme 74 Step 1: Intermediate 74-a To a solution of intermediate 50-c (300 mg, 0.89 mmol) and intermediate 30-b (229 mg, 0.97 mmol) in 1,4-dioxane (890 pl) were sequentially added N,N-dimethylglycine (275 mg, 2.67 mmol), copper(I) iodide (169 mg, 0.89 mmol) and cesium carbonate (870 mg, 2.67 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate.
The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided intermediate 74-a=HCI as beige solid.
Step 2: Compound 130 A solution of intermediate 74-a (125 mg, 0.25 mmol), trimethyl orthoformate (836 pl, 7.65 mmol) and PTSA (catalytic) was stirred at room temperature for 3 hours. Excess trimethyl orthoformate was removed in vacuo and the residue was treated with ammonia (7.0 N in Me0H) (2.55 ml, 5.10 mmol).
The mixture was stirred at room temperature for 2 days and volatiles were removed under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 130.2HCI as a white solid. MS (m/z) M+H= 518.1 Synthesis of Compound 97:
F F
fa Br 1 ?I O= o, it --=.-\ 0 ----\-NC Cul, Cs2CO3 HC(OMe)3 NH2 iii -.........*. C _____________________ I.
/ \ \/N , \ /
H2N V'm 36-f I ,N CNNo I NH3 ,N Co N N
)\ HN2N N)Th N
U
0 0 \--0) irk 43-e 75-a Compound 97 Scheme 75 Step 1: Intermediate 75-a To a solution of intermediate 43-e (101 mg, 0.29 mmol) and intermediate 36-f (110 mg, 0.32 mmol) in 1,4-dioxane (1.4 ml) were sequentially added N,N-dimethylglycine (90 mg, 0.87 mmol), copper(I) iodide (56 mg, 0.29 mmol) and cesium carbonate (381 mg, 1.17 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate.
The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 75-a as a beige solid.
Step 2: Compound 97 Intermediate 75-a (160 mg, 0.26 mmol), trimethyl orthoformate (1.86 ml, 17.09 mmol) and TFA (catalytic) were heated at 110 C for 1 hour. Excess trimethyl orthoformate was removed in vacuo and the residue was treated with 7.0 N ammonia in Me0H (1.87 ml, 13.14 mmol). The mixture was stirred at 50 C overnight and volatiles were removed under reduced pressure. Purification by reverse phase chromatography eluting with a 1 70 aqueous HCl/methanol gradient provided compound 97.2HCI as a beige solid.
MS (m/z) M+H= 636.1 Synthesis of Compound 99:
F F
0 46 0 .
NH2 OH2 Pd/C NH2 fit ---=--\-_____________________________________ ).
NV r "N N ,N NV i 1 "N N N
I ,) o a * 0 Compound 97 Compound 99 Scheme 76 A solution of compound 97.2HCI (100 mg, 0.15 mmol) in ethyl acetate was treated with 10% palladium on carbon (32 mg, 0.015 mmol) and purged with H2. The solution was stirred under H2(1 atm) for 1 hour before being filtered through celite. The filtrate was concentrated in vacuo. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 99.2HCI as white solid. MS (m/z) M+H= 546.2 Synthesis of Compound 93:
F F
Br 1 9 0 * o 0 *
NJ-OH
NC 41 Cul, Cs2CO3 4. -------% *
___________________ w NC N - + NC N.--,,/N---/
H2N N-- 37-f + 37-f 1 \,N r , \,N
H2N )_____\ 1 N
\-SiMe3 -0) 43-e 77-a 77-a' F
0.
F
0 ft HC(OMe)3 NH2 * -":---"\--- NH2 *
77-a + 77-a' --1.- N / N + , -_/
NH3 N "*" i " N N
L 1 N r -z N ' \
l'i, o N' SiMe3 77-h 77-h' F
O, HCI NH2 * 0 ---\-- õ, 77-h + 77-b' ---0,- / HNrµ
N
l " ,N
N l'I)_Th U
Compound 93 Scheme 77 Step 1: Intermediates 77-a and 77-a' To a solution of intermediate 43-e (115 mg, 0.33 mmol) and intermediates 37-f and 37-f' (123 mg, 0.36 mmol) in 1,4-dioxane (1.6 ml) were sequentially added N,N-dimethylglycine (102 mg, 0.99 mmol), copper(I) iodide (63 mg, 0.33 mmol) and cesium carbonate (431 mg, 1.32 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate. The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 77-a and 77-a' as an inseparable mixture.
Step 2: Intermediates 77-b and 77-b' A solution of intermediates 77-a and 77-a' (100 mg, 0.16 mmol), trimethyl orthoformate (2.0 ml, 18.28 mmol) and PTSA (catalytic) was stirred at room temperature for 1 hour. Excess trimethyl orthoformate was removed in vacuo and the residue was treated with 7.0 N ammonia in Me0H (2.0 ml, 14.1 mmol). The mixture was stirred at room temperature for 3 days and volatiles were removed under reduced pressure. Purification by silica gel chromatography provided intermediates 77-b and 77-b' as an inseparable mixture.
Step 3: Compound 93 4N HCI in 1,4-dioxane (2 mL) was added to intermediates 77-b and 77-b' (60 mg, 0.09 mmol) band the mixture was stirred for 1 hour at room temperature. Volatiles were removed under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 93.2HCI as a white solid. MS (m/z) M+H= 502.2 Synthesis of Compound 118:
F F
Br 1 On 0 * 0 .
NC 11 Cut, Cs2CO3 *--------\-- *
/ \
___________________ ).
N NC r-.N m e'' + NC \
I
H2N N-- 38-e +38-e' 1 ",N 1 1 N
a N 0 H2N __ )__\ 1 SiMe3 N
\
0 --0) \---0) 43-e 78-a 78-a' F F
0 10 0 .
HC(OMe)3 NH2 4* -----\--- n, + NH2 4* ,---=\-(:)/"SiMe3 78-a + 78-a' 0- N r II N.--/
NH3 N ' , \
r '( N" \
I N'N NK \
N Q t`l..Th I N
SiMe3 78-b 78-b' F
0 .
HCI NH2 . ------'\
78-b + 78-b' 1. ' HN ,,, N
N \
Th\I r4)____I
Compound 118 Scheme 78 Step 1: Intermediates 78-a and 78-a' To a solution of intermediate 43-e (276.0 mg, 0.79 mmol) and intermediates 38-e and 38-e' (340 mg, 0.87 mmol) in 1,4-dioxane (795 pl) were sequentially added N,N-dimethylglycine (246 mg, 2.38 mmol), copper(I) iodide (151 mg, 0.79 mmol) and cesium carbonate (1.03 g, 3.18 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate. The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 78-a and 78-a' as an inseparable mixture.
Step 2: Intermediates 78-b and 78-b' A solution of intermediates 78-a and 78-a' (90 mg, 0.14 mmol), trimethyl orthoformate (1.03 ml, 9.45 mmol) and TFA (catalytic) was stirred at room temperature for 1 hour. Excess trimethyl orthoformate was removed in vacuo and the residue was treated with 7.0 N ammonia in Me0H (1.03 ml, 7.27 mmol). The mixture was stirred at room temperature overnight and volatiles were removed under reduced pressure to provide intermediates 78-b and 78-b' as an inseparable mixture.
Step 3: Compound 118 4N HCI in 1,4-dioxane (2.82 mL) was added to intermediates 78-b and 78-b' (60 mg, 0.09 mmol) and the mixture was stirred overnight at room temperature. Volatiles were removed under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 118.2HCI as a white solid. MS (m/z) M+H=
516.2 Synthesis of Compound 90:
F F
Br 1 ?I 0O 0 fa 41, NC Cul, Cs2CO3 0HC(OMe)3 NH2 * ---\----/ \
NC ,N,.// NH3 N ''. \
H2N N'N 34-d 1 \ N L I N
H N
H2N N\
M
OH OH OH
47-a 79-a Compound 90 Scheme 79 Step 1: Intermediate 79-a To a solution of intermediate 79-a (310 mg, 1.0 mmol) and intermediate 34-d (313 mg, 1.21 mmol) in 1,4-dioxane (2.5 ml) were sequentially added N,N-dimethylglycine (156 mg, 1.51 mmol), copper(I) iodide (96.0 mg, 0.50 mmol) and cesium carbonate (1.31 g, 4.04 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate. The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided intermediate 79-a as a beige solid.
Step 2: Compound 90 Intermediate 79-a (280 mg, 0.62 mmol) and trimethyl orthoformate (2.05 ml, 18.7 mmol) were heated at 110 C for 3 hours. Excess trimethyl orthoformate was removed in vacuo and the residue was treated with 7.0 N
ammonia in Me0H (1.78 ml, 12.5 mmol). The mixture was stirred at room temperature overnight and volatiles were removed under reduced pressure.
Purification by reverse phase chromatography eluting with a 1 /0 aqueous HCl/methanol gradient provided compound 90.2HCI as a beige solid. MS
(m/z) M+H= 476.2 Synthesis of Compound 103:
F F
Br 1 0 0 * AO o O*
NC Cul, Cs2CO3 fl Cr---:\ HC(OMe)3 NH2 . -.---;--\,, _________________________________________________ s N / N
/ \ NC ,...- ---./ NH3 N , \
H2N N'N 34-d =
1 N L I ,N
H2N o ,N N)._..., U
46-a 80-a Compound 103 Scheme 80 Step 1: Intermediate 80-a To a solution of intermediate 46-a (300 mg, 0.87 mmol) and intermediate 34-d (247 mg, 0.95 mmol) in 1,4-dioxane (2.1 ml) were sequentially added N,N-dimethylglycine (134 mg, 1.30 mmol), copper(I) iodide (83 mg, 0.43 mmol) and cesium carbonate (1.13 g, 3.48 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated and the aqueous phase was extracted twice with ethyl acetate. The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided intermediate 80-a=FICI as a beige solid.
Step 2: Compound 103 Intermediate 80-a=HCI (65.0 mg, 0.13 mmol) and trimethyl orthoformate(3.0 ml, 4.01 mmol) were heated at 110 C for 3 hours. Excess trimethyl orthoformate was removed in vacuo and the residue was treated with 7.0 N
ammonia in Me0H (378 pl, 2.65 mmol). The mixture was stirred at room temperature overnight and volatiles were removed under reduced pressure.
Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 103.2HCI as a white solid. MS
(m/z) M+H= 514.2 Synthesis of Compound 120:
Br O * 410 o *
NC Cul, Cs2003 = HC(OMe)3 NH2 10 N
N
HN N NC NH3'N 35-d i I JA
\,N1 42-d 81-a Compound 120 Scheme 81 Step 1: Intermediate 81-a To a solution of intermediate 42-d (176 mg, 0.53 mmol) and intermediate 35-d (145 mg, 0.53 mmol) in 1,4-dioxane (3.5 ml) were sequentially added N,N-dimethylglycine (123 mg, 1.19 mmol), copper(I) iodide (76 mg, 0.40 mmol) and cesium carbonate (693 mg, 2.13 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated, and the aqueous phase was extracted twice with ethyl acetate.
The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided intermediate 81-a.HCI as a yellow foam.
Step 2: Compound 120 Intermediate 81-a=HCI (259 mg, 0.53 mmol) and trimethyl orthoformate (3.78 ml, 34.6 mmol) were heated at 110 C for 1 hour. Excess trimethyl orthoformate was removed in vacuo and the residue was treated with 7.0 N
ammonia in Me0H (3.80 ml, 26.6 mmol). The mixture was heated at 60 C
for 5 hours and volatiles were removed under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 120.2HCI as a white solid. MS (m/z) M+H=
514.2 Synthesis of intermediate 82-c:
Br Br Br 11 0 DIPEA =
TMS-diazomethane =CI malononitrile CN CN
HO Me0 CN CN
82-a 82-b 82-c Scheme 82 Step 1: Intermediate 82-b To a solution of 4-bromo-2-fluorobenzoyl chloride (16.27 g, 68.5 mmol) in toluene (85 ml) and THF (8.5 ml), cooled to -10 C, were sequentially added malononitrile (4.75 g, 71.9 mmol) and DIPEA (23.93 ml, 137.0 mmol) in toluene (25 mL) drop wise over a period of 1 hour. After the addition was completed, the reaction was stirred for 2 hours at 0 C and room temperature for an additional 2 hours. Volatiles were removed under reduced pressure.
1N HCI and ethyl acetate were added to the residue; the organic layer was separated, washed twice with 1N HCI and brine, dried over MgSO4, filtered and concentrated under reduced pressure to provide intermediate 82-b as a yellow solid.
=
Step 2: Intermediate 82-c To a solution of intermediate 82-b (18.29 g, 68.5 mmol) in acetonitrile (247 ml) and methanol (27.4 ml), cooled to 0 C, was added DIPEA (14.36 ml, 82.0 mmol) and a 2M solution of diazomethyl)trimethylsilane in hexanes (37.7 ml, 75.0 mmol). After the addition was completed, the reaction was stirred at room temperature overnight. Acetic acid (1.17 ml, 20.5 mmol) was added, the reaction was stirred for 30 minutes and volatiles were removed under reduced pressure. A saturated aqueous solution of NaHCO3 and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure.
Purification by silica gel chromatography provided intermediate 82-c as a yellow solid.
Synthesis of Intermediate 83-a:
Br Br TEA
441k NC
NC 43-d 0 \ N
82-c 83-a Scheme 83 To a solution of intermediate 82-c (2.0 g, 7.12 mmol) and TEA (1.98 ml, 14.23 mmol) in Et0H (3.50 ml) was added intermediate 43-cl=FICI (1.30 g, 8.54 mmol) and the reaction was then stirred for 2 hours at 100 C. Volatiles were removed under reduced pressure. A saturated aqueous solution of ammonium chloride and ethyl acetate were added to the residue, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure to provide intermediate 83-a as a yellow solid.
Synthesis of Compound 60:
Br 0 I. 0 0 =
NH2 41k s,õN
NC F Cul, Cs2CO3 Formamide I "'N NC N N
N 1_ 29-i 83-a 84-a Compound 60 Scheme 84 Step 1: Intermediate 84-a To a solution of intermediate 83-a (2.60 g, 7.12 mmol) and intermediate 29-1 (1.00 g, 4.18 mmol) in 1,4-dioxane (20.9 ml) were sequentially added N,N-dimethylglycine (646 mg, 6.27 mmol), copper(I) iodide (398 mg, 2.09 mmol) and cesium carbonate (4.09 g, 12.5 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated, and the aqueous phase was extracted twice with ethyl acetate. The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 84-a as a beige solid.
Step 2: Compound 60 Formamide (11.7 ml, 293 mmol) was added to intermediate 84-a (2.18 g, 4.18 mmol) and the reaction was stirred at 180 C overnight. Water and ethyl acetate were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure.
Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 60.2HCI as white solid. MS (m/z) M+H= 551.2 Synthesis of Compound 73:
F
F
Br 0 el 0 0 =
F /N---z/ NH2 . --)---\
1\1 N
NC
Cul, Cs2CO3 HC(OMe)3 F F
_________________ ).
I N m ' _________________ N1LT
\ NC N
1 \,N
\ Nr NH3 H2N N 34-d y..,.\ H2N N
(--()I
83-a 85-a Compound 73 Scheme 85 Step 1: Intermediate 85-a To a solution of intermediate 83-a (2.60 g, 7.12 mmol) and intermediate 34-d (1.0 g, 4.18 mmol) in 1,4-dioxane (20.9 ml) were sequentially added N,N-dimethylglycine (646 mg, 6.27 mmol), copper(I) iodide (398 mg, 2.09 mmol) and cesium carbonate (4.09 g, 12.5 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated, and the aqueous phase was extracted twice with ethyl acetate. The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 85-a as a beige solid.
Step 2: Compound 73 Intermediate 85-a (195.0 mg, 0.38 mmol) and trimethyl orthoformate (1.26 ml, 11.59 mmol) were heated at 110 C for 3 hours. Excess trimethyl orthoformate was removed in vacuo and the residue was treated with 7.0 N
ammonia in Me0H (3.86 ml, 7.72 mmol). The mixture was stirred at room temperature overnight and volatiles were removed under reduced pressure.
Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 73.HCI as white solid. MS (m/z) M+H= 534.1 Synthesis of Compound 95:
F
F
Br 0 el 0 0 .
4k ,NIOH 40 1 N
/N
NC F Cul, Cs2CO3 __________________ > F ., HC(OMe)3 __________________________________________________ N . O_:21 " F
I \, H2N N NC ,,, 1 31-d \ Nr. [,,, ,N
N
11)____, (--)1, 83-a 86-a Compound 95 Scheme 86 Step 1: Intermediate 86-a To a solution of intermediate 83-a (200 mg, 0.54 mmol) and intermediate 31-d (177 mg, 0.65 mmol) in 1,4-dioxane (3.65 ml) were sequentially added N,N-dimethylglycine (127 mg, 1.23 mmol), copper(I) iodide (7.8 mg, 0.41 mmol) and cesium carbonate (714 mg, 2.19 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated, and the aqueous phase was extracted twice with ethyl acetate.
The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided intermediate 86-a=HCI as a beige solid.
Step 2: Compound 95 Intermediate 86-a=HCI (284 mg, 0.54 mmol) and trimethyl orthoformate (3.90 ml, 35.6 mmol) were heated at 110 C for 1 hour. Excess trimethyl orthoformate was removed in vacuo and the residue was treated with 7.0 N
ammonia in Me0H (3.91 ml, 27.4 mmol). The mixture was stirred at room temperature overnight and volatiles were removed under reduced pressure.
Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 95.2HCI as white solid. MS (m/z) M+H= 545.2 Synthesis of Intermediate 87-a:
Br Br ,.
F NC F
NC 42-c 0 I \ N
82-c 87-a a Scheme 87 To a solution of intermediate 82-c (1.00 g, 3.56 mmol) and TEA (1.09 ml, 7.83 mmol) in Et0H (3.50 ml) was added intermediate 42-c (583 mg, 4.27 mmol) and the reaction was then stirred for 2 hours at 100 C. Volatiles were removed under reduced pressure. A saturated aqueous solution of ammonium chloride and ethyl acetate were added to the residue, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 87-a as beige solid.
Synthesis of Compound 92:
F
F
Br 0 el 0 0 =
. ''N -)LOH 40 r\,N
/N--Zi NH2 10 NC F Cul, Cs2CO3 F
_________________ 1.- HC(OMe)3 F
NC "
I \,N K1 NH3 34-d \ Nr L I pi H2N f \l,..,_ N NL
c-J H2N 0 U
87-a 88-a Compound 92 Scheme 88 Step 1: Intermediate 88-a To a solution of intermediate 87-a (452 mg, 1.29 mmol) and intermediate 34-d (368 mg, 1.42 mmol) in 1,4-dioxane (5.20 ml) were sequentially added N,N-dimethylglycine (400 mg, 3.88 mmol), copper(I) iodide (247 mg, 1.29 mmol) and cesium carbonate (1.26 g, 3.88 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated, and the aqueous phase was extracted twice with ethyl acetate. The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided intermediate 88-a=HCI as a beige solid.
Step 2: Compound 92 Intermediate 88-a=HCI (124.0 mg, 0.25 mmol) and trimethyl orthoformate (935 pi, 7.63 mmol) were heated at 110 C for 3 hours. Excess trimethyl orthoformate was removed in vacuo and the residue was treated with 7.0 N
ammonia in Me0H (2.54 ml, 5.09 mmol). The mixture was stirred at room temperature overnight and volatiles were removed under reduced pressure.
Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 92.2HCI as a beige solid. MS
(m/z) M+H= 518.2 Synthesis of intermediate 89-a:
Br Br _____________________________________________________ , *
F TEA NC F
NC 44-c 0 l \ N
)----82-c 89-a Scheme 89 To a solution of intermediate 82-c (2.0 g, 7.12 mmol) and TEA (2.18 ml, 15.65 mmol) in Et0H (7.12 ml) was added intermediate isopropylhydrazine hydrochloride (944 mg, 8.54 mmol) and the reaction was then stirred for 2 hours at 100 C. Volatiles were removed under reduced pressure. A
saturated aqueous solution of ammonium chloride and ethyl acetate were added to the residue, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure.
Purification by silica gel chromatography provided intermediate 89-a as a beige solid.
Synthesis of Compound 98:
F
F
Br 0OO 0 =
iii .--14).1''ON 40 Cr\,N
F
F /N --S HC(OMe)3 NH2 . .----\-- m NC Cul, Cs2CO3 , N F
l \,N NC õI 1 \ NH3 i_ ,N
34-d N :. N\
H2N Nj H2N---1."--89-a 90-a Compound 98 Scheme 90 Step 1: Intermediate 90-a To a solution of intermediate 89-a (400 mg, 1.24 mmol) and intermediate 34-d (352 mg, 1.36 mmol) in 1,4-dioxane (4.9 ml) were sequentially added N,N-dimethylglycine (383 mg, 3.71 mmol), copper(I) iodide (236 mg, 1.24 mmol) and cesium carbonate (1.21 g, 3.71 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated, and the aqueous phase was extracted twice with ethyl acetate. The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided intermediate 90-a=FICI as beige solid.
Step 2: Compound 98 Intermediate 90-a=HCI (208 mg, 0.45 mmol) and trimethyl orthoformate (1.47 ml, 13.4 mmol) were heated at 110 C for 3 hours. Excess trimethyl orthoformate was removed in vacuo and the residue was treated with 7.0 N
ammonia in Me0H (4.48 ml, 8.96 mmol). The mixture was stirred at room temperature for 3 days and volatiles were removed under reduced pressure.
Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 98.2HCI as a beige solid. MS
(m/z) M+H= 492.2 Synthesis of Intermediate 91-d:
=
Br Br Br Br F F is F 0 oxalyl chloride F 0 ________________ ).- malononitrile =TMS-diazomethane NaOH
CN CN
HO '=-= Me0 0 OH 0 Cl CN CN
91-a 91-b 91-c 91-d Scheme 91 Step 1: Intermediate 91-b To a suspension of 4-bromo-3-fluorobenzoic acid (15.0 g, 68.5 mmol) in dichloromethane (342.0 ml) was added DMF (106.0 pl, 1.37 mmol) and oxaly1y1 chloride (8.99 ml, 103.0 mmol). The solution was then stirred at room temperature for 3 hours. Volatiles were removed under reduced pressure to provide intermediate 91-b as a yellow solid.
Step 2: Intermediate 91-c To a solution of intermediate 91-b (16.27 g, 68.5 mmol) in toluene (85.0 ml) and THF (8.5 ml) cooled to -10 C were sequentially added malononitrile (4.75 g, 71.9 mmol) and DIPEA (23.93 ml, 137 mmol) drop wise over a period of 15 minutes. The reaction was stirred at 0 C for 2 hours and room temperature for an additional 2 hours. Volatiles were removed under reduced pressure. Ethyl acetate and 1N HCI were added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated in vacuo to provide intermediate 91-c as a yellow solid.
Step 3: Intermediate 91-d To a solution of intermediate 91-c (18.29 g, 68.5 mmol) in acetonitrile (247.0 ml) and Me0H (27.4 ml) cooled to 0 C were sequentially added DIPEA (14.36 ml, 82.0 mmol) and a 2M solution of TMS-Diazomethane in hexanes (37.7 ml, 75.0 mmol). The reaction was then stirred for 5 hours at room temperature. Acetic acid (1.17 ml, 20.55 mmol) was then added and the reaction was stirred for an additional 30 minutes. Volatiles were removed under reduced pressure. A saturated aqueous solution of NaHCO3 and ethyl acetate was added, the organic layer was separated, washed with brine, dried over MgSO4, filtered and concentrated in vacuo. Purification by silica gel chromatography provided intermediate 91-d as a beige solid.
Synthesis of Intermediate 92-a:
Br Br F
TEA
NC NC
43-d CN
H2N I \N
91-d 92-a Scheme 92 To a solution of intermediate 91-d (2.0 g, 7.12 mmol) and TEA (1.98 ml, 14.23 mmol) in Et0H (3.50 ml) was added intermediate 43-d.HCI (1.30 g, 8.54 mmol) and the reaction was then stirred for 2 hours at 100 C. Volatiles were removed under reduced pressure. A saturated aqueous solution of ammonium chloride and ethyl acetate were added to the residue, the organic layer was separated, washed with brine, dried over MgSO4, ffitered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 92-a as a beige solid.
Synthesis of Compound 59:
Br 0 I. 0 0 j=LOH
8,711 Cul, Cs2CO3 Formamide NH2 NC
I "'NI NC \"N
__________________________________________________ N
29-i N I ,N
N
-12N N)..Th 92-a 93-a Compound 59 Scheme 93 Step 1: Intermediate 93-a To a solution of intermediate 92-a (2.60 g, 7.12 mmol) and intermediate 29-1 (1.0 g, 4.18 mmol) in 1,4-dioxane (20.90 ml) were sequentially added N,N-dimethylglycine (646 mg, 6.27 mmol), copper(I) iodide (398 mg, 2.09 mmol) and cesium carbonate (4.09 g, 12.54 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated, and the aqueous phase was extracted twice with ethyl acetate. The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 93-a as a beige solid.
Step 2: Compound 59 Formamide (11.67 ml, 293.0 mmol) was added to intermediate 93-a (2.18 g, 4.18 mmol) and the reaction was stirred at 180 C overnight then cooled to room temperature. Water and ethyl acetate were added, the organic layer was separated, washed with brine, dried over Mg504, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 59.2HCI as a yellow solid. MS (m/z) M+H= 551.1 Synthesis of Compound 72:
F
F
Br 0 Si 0 o =
th F -7`0H
N-(/ .- it NH, ,..N N
NC
Cul, Cs2CO3 HC(OMe)3 i "N NC N m 34-d \ NI NH3 N)_Th (---1;) 92-a 94-a Compound 72 Scheme 94 Step 1: Intermediate 94-a To a solution of intermediate 92-a (291.0 mg, 0.79 mmol) and intermediate 34-d (227.0 mg, 0.87 mmol) in 1,4-dioxane (3.2 ml) were sequentially added N,N-dimethylglycine (247.0 mg, 2.39 mmol), copper(I) iodide (152.0 mg, 0.79 mmol) and cesium carbonate (780 mg, 2.39 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated, and the aqueous phase was extracted twice with ethyl acetate.
The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided intermediate 94-a=FICI as a beige solid.
Step 2: Compound 72:
Intermediate 94-a=HCI (56.0 mg, 0.11 mmol) and trimethyl orthoformate (363 pl, 3.32 mmol) were heated at 110 C for 3 hours. Excess trimethyl orthoformate was removed in vacuo and the residue was treated with 7.0 N
ammonia in Me0H (1.10 ml, 2.21 mmol). The mixture was stirred at room temperature overnight and volatiles were removed under reduced pressure.
Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 72.2HCI as a white solid. MS
(m/z) M+H= 534.2 Synthesis of Compound 68:
F
Br 0 411t , 0=
0--......, , \N
NH2 4410 N)-LOH NH2 *
---j N \ Cul, Cs2CO3 N.:
N
l , _____________ 0, N'"N
N N)_\ I , \--__) .
0 39-b N
U
17-a Compound 68 Scheme 95 To a solution of intermediate 17-a (300.0 mg, 0.80 mmol) and intermediate 39-b (177.0 mg, 0.80 mmol) in 1,4-dioxane (3.6 ml) were sequentially added N,N-dimethylglycine (372.0 mg, 3.61 mmol), copper(I) iodide (229.0 mg, 1.20 mmol) and cesium carbonate (1.04 g, 3.21 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated, and the aqueous phase was extracted twice with ethyl acetate.
The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 68.2HCI as a beige solid. MS (m/z) M+H= 514.2 Synthesis of Compound 69:
F
Br 0 .
----)r-N
NH2 * N).LOH NH2 40 N\j,_ N \ N Cul, Cs2CO3 I, \
N N)__\ L I ,N
U0 40-b N
) 17-a Compound 69 Scheme 96 To a solution of intermediate 17-a (300.0 mg, 0.80 mmol) and intermediate 40-b (177.0 mg, 0.80 mmol) in 1,4-dioxane (3.6 ml) were sequentially added N,N-dimethylglycine (372.0 mg, 3.61 mmol), copper(I) iodide (229.0 mg, 1.20 mmol) and cesium carbonate (1.04 g, 3.21 mmol). The reaction mixture was stirred at reflux overnight, cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated, and the aqueous phase was extracted twice with ethyl acetate.
The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% aqueous HCl/methanol gradient provided compound 69.2HCI as a beige solid. MS (m/z) M+H= 514.2 Synthesis of Compound 63:
Br NH2 Os0 111j-LOH 0 INV \ Cul, Cs2CO3 1 'N N
N 1 'N
0 41-b N
17-a Compound 63 Scheme 97 A solution of intermediate 17-a (437 mg, 1.17 mmol), intermediate 11-c (255 mg, 1.17 mmol), quinolin-8-ol (34 mg, 0.23 mmol), copper (I) iodide (44.0 mg, 0.23 mmol) and cesium carbonate (761 mg, 2.33 mmol), in dimethylacetamide (1.2 ml), was degassed with argon for 10 minutes, heated in a sealed tube at 140 C overnight and then cooled to room temperature. Water and ethyl acetate were added, the organic layer was separated, the aqueous layer was extracted twice with ethyl acetate, the combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% HCl/methanol gradient provided compound 63.1-1CI as a yellow solid. MS (m/z) M+H= 512.2 Synthesis of Compound 67:
N
i F0 Br K2CO3 F Br Cul, DEA, Pd(PPh3)2Cl2 F 0 NI\
_________________ * __ MOMCI Vi 1 Nr ----_:-----XN1 OH OMOM OMOM
I
98-a 98-b 98-c 98-d N N
F I F i H2 Pd/C N HCI
98-d ______________ )= 1 OMOM OH
98-e 98-f F
Br NH2 41* N / N
..--- -...../
NI ' \N Cul I' N NI)Th . I ,N
98-f N NI)______\
S---.0) U
17-a 0 compound 67 Scheme 98 Step 1: Intermediate 98-b To a solution of 3-bromo-5-fluorophenol, 98-a (25.00 g, 131.0 mmol), in acetone (654 ml) were sequentially added K2CO3 (27.10 g, 196.0 mmol) and chloro(methoxy)methane (11.59 g, 144.0 mmol). The reaction was stirred at room temperature for 2 hours and then filtered. The filtrate was concentrated under reduced pressure to provide intermediate 98-b as a yellow oil.
Step 2: Intermediate 98-d To a solution of intermediate 98-b (2.00 g, 8.51 mmol) in DMF (17.02 ml), were sequentially added diethylamine (975 pl, 9.36 mmol), copper(I)iodide (65 mg, 0.34 mmol) and 5-ethyny1-1-methyl-1H-imidazole 98-c (948 mg, 8.93 mmol). After copper(I)iodide has completely dissolved, Dichlorobis(triphenylphosphine)palladium(II) (119 mg, 0.17 mmol) was added and the reaction was then stirred at 100 C overnight and then cooled to room temperature. Water and ethyl acetate were added, the organic layer was separated, the aqueous layer was extracted twice with ethyl acetate, the combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by silica gel chromatography provided intermediate 98-d as a beige solid Step 3: Intermediate 98-e A solution of intermediate 98-d (600.0 mg, 2.3 mmol) in methanol was treated with 10% palladium on carbon (245.0 mg, 0.01 mmol) and purged with H2. The solution was stirred under H2 (1 atm) overnight before being filtered through celite. The filtrate was concentrated in vacuo to provide intermediate 98-e as a yellow oil.
Step 4: Intermediate 98-f To a solution of intermediate 98-e (2.4 g, 9.08 mmol) in Me0H (17 ml) was added 4N HCI in dioxane (2.76 ml, 91 mmol). The reaction mixture was stirred at room temperature for 2 hours. Volatiles were removed under reduced pressure, diethyl ether was added to the residue; a precipitate formed and was collected by filtration to provide intermediate 98-f=HCI as a white solid.
Step 5: Compound 67 A solution of intermediate 17-a (292 mg, 0.78 mmol), intermediate 98-f=HCI
(200 mg, 0.78 mmol), tetramethylheptane-3,5-dione (287 mg, 1.56 mmol), copper (I) iodide (148 mg, 0.78 mmol) and cesium carbonate (762 mg, 2.33 mmol), in NMP (3.9 ml), was degassed with argon for 10 minutes, heated in a sealed tube at 120 C overnight and then cooled to room temperature, diluted with ethyl acetate and filtered over celite. A saturated aqueous solution of ammonium chloride was added to the filtrate, the organic layer was separated, and the aqueous phase was extracted twice with ethyl acetate. The combined organic extracts were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. Purification by reverse phase chromatography eluting with a 1% HCl/methanol gradient provided compound 67.2HCI as a yellow solid. MS (m/z) M+H= 514.3 Compounds 7, 8, 11, 13, 14, 19, 21, 24 to 28, 33, 34, 37 to 51, 53, 56, 61, 62, 64, 66, 70, 71, 74-77, 79 to 84, 86 to 89, 94, 96, 104, 105, 107 to 112, 115, 116, 119, 121, 122, 123, 124, 126, 127, 131, 132, and 133 were prepared using similar methods to those described above.
Table 1 summarizes representative compound of Formula 1.
Table 1: Example Compounds of Formula 1 Compound Structure MS (m/z) o, 1 NH2 * [M+H]=410.2 N.-- \ O
1 ,N
N N
H
o, NH2 4It 40 [M+H]=478.2 N' 1 \
I N
N N),õ_, U
o, NH2 *
0[M+H]=593.1 3 N \
L 1 ,N
NN,_____ e 0--.
o, NH2 *
* [M+H]=493.1 N" \N
N N, òH
o, NH2 *
N" * [M+H]=547.1 I \N
' N N
o, NH2 44*
6 fh[M+H]=535.1 N -' \ N
I ' N N
o, NH2 4.
O[M+Hr=563.1 N N
----\
0 =
NH2 4k 8 N '' , \ O [M+H]=563.1 I ,N
N N
L\N---e i----, NH2 4.
=
9 ci [M+H]=512.2 N \
I
N N
o, NH2 it [M+H]=508.1 , \ ¨0 I ,N
N N
*
11 0, [M+H]=508.2 \N
I ' N
o*
CN
NH2 41i =
12 [M+H]=503.2 N' \N
N N
o, NH2. .
13 [M+Hr=478.2 N \N
I ' N NL
U
0 =
NH2 44* S N
14 [M+Hr=485.2 N' \N
I ' U
o, NH2. .
15 [M+Hr=503.3 N' \N
l , N NL
U
o, 16 NH2 0 fb [M+H]=424.2 N' \N
l , N N
\
o, CN
17 NH2 fi fa [M+H]=449.3 N' , \
L 1 ,N
1\1 N
\
o,
18 NH2 40 0--)..______.
[M+H]=445.1 N
N' \N
N l N' \
o =
om_-.//____\
[M+H]=445.1 N
N' \N
N l N' \
o =
om_-.//____\
19 NH2 4. N
S- [M+H]=431.4 N" , \
L 1 ,N
r\I NI
\
o,
S- [M+H]=431.4 N" , \
L 1 ,N
r\I NI
\
o,
20 NH2 44, 401 [M+H]=492.1 N", \
L 1 ,N
'N N
\
o, NH2 .
L 1 ,N
'N N
\
o, NH2 .
21 [M+H]=494.2 N \N
I , N
U
o, CN
NH2 4.
I , N
U
o, CN
NH2 4.
22 N [M+Hr=519.2 ' \
L I ,N
N )....ThN
U
o, NH2 4. 40
L I ,N
N )....ThN
U
o, NH2 4. 40
23[M+H]=562.1 N' \N
I , N N)_Th U
NH2 *
I , N N)_Th U
NH2 *
24 [M+H]=501.2 ,N
N N
0 =
N N
0 =
25 N \N [M+H]=560.2 ' N N1) UTh( 0 = CF3 NH2 44*
26 \ N [M+H]=603.1 I , N
0 =
OTh-S
NH2 \N*
0 =
OTh-S
NH2 \N*
27 N' \ N [M+H]'=542.2 I' N N_ UN5( O * CN
NH2 O41Ik
NH2 O41Ik
28 N' \ N [M+H]4=560.2 I' N N3.
Urs1( O fal CF3 NH2 * *
Urs1( O fal CF3 NH2 * *
29 [M+H]'=546.2 N' \
U
o, CN
U
o, CN
30 NH2 ii O [M+H]4=449.4 N' , \
1 ,N
N N
\
o,
1 ,N
N N
\
o,
31 NH2 * 44Ik [M+H]=492.1 N' \N
1 , N N
\
o, NH2 .
44k
1 , N N
\
o, NH2 .
44k
32 [M+H]=546.1 N' \
1 ,N
N N
b o, NH2 =
=
1 ,N
N N
b o, NH2 =
=
33 NJ' \
l [M+Hr=603.1 L ,N
N N
oN-{
o, NH2 * S N
l [M+Hr=603.1 L ,N
N N
oN-{
o, NH2 * S N
34 [M+H]=488.3 N' \N
1.l , N N\
-----\
N---o, NH2. NC 44Ik
1.l , N N\
-----\
N---o, NH2. NC 44Ik
35 [M+H]=519.2 \
I ,N
N N' K) o, NH2 * F3C =
I ,N
N N' K) o, NH2 * F3C =
36 [M+H]=562.2 N \N
I' N 14),Th o, NH2 441k
I' N 14),Th o, NH2 441k
37 N [M+Hr=499.1 N' \N
1 ' c----i 44k
1 ' c----i 44k
38 [M+Hr=515.1 \
L ,N
ymN
fi
L ,N
ymN
fi
39 [M+Hr=514.2 ,N
Nv_ o, L ,N [M+Hr=543.1 (N--) \--N1 0--)1 NH2 41*
41 N \N [M+Hr=522.1 N' o, 42 N \ N [M+H]=521.2 N#
0¨\
( µj 43 [M+H]=500.2 ,N
N
o, NH2 0--\ 4* US) NI/
44 NI \ [M+H]=530.1 I N
' N Niv -------\
--C) o, 45 NH2 0 40 [M+H]=452.1 NI' \N
I ' N N
/'0 F
o, 46 NH2 44k N [M+H]=519.1 N' \
/ N
' N NI)._.....\
fi ) S
L ,N [M+Fi]=542.1 1%1 /C) fk 48 [M+Hr=498.2 N
,N
N N)_Th 0 =
49 , [M+H]=600.1 L ,N
Thµl r=L
0-<
4Ik , S
50 NH2 =[M+H]=503.2 r%V \
,N
N
o, NH2 Os 51 [M+H]=500.2 N
' N N
L\NH
o, 52 [M+H]=562.1 N'N
I' N N\
\-0 o, OTh---N
53[M+H]=544.2 N'' \N
I' N rµI___ F
0 =
NH2. --==---- \-sN/1µ1 [M+Hr=533.1 /
N- \
l N1 , 1 N N)___\
---'0) F
o, OTh-\--SK./N
[M+H]=576.2 N.-- \N
1' N N___ -\----N3 F
o, 0-Al__ NH2 . N [M+H1=532.2 N' N 1 ' N N
- F
o, NH2 41*
[M+H]=546.2 57 \
NV \N
1 ' N N\
----\
Nc13 F
o, OTh.,__.\
N1 [M+H]=517.2 =
N"
1 ' _.---1 *
NH2 40. SN
[M+Hr=551.1 \N
' N Nym O*
[M+H]=551.2 N" \N
' N N)Th o*
61NH2 O[M+H]=463.2 S,,K/N
N" \
N'N
0 4Ik F
0---\-SK/
NH2 441, N
1 [M+H]=533.2 N' \N
I' N Ny.Th --- )0 F
o, NH2 40 gli [M+H]=512.2 N' \
I N
' N NI)...Th U
F
0 . .V
S/
NH2 41* 1\1 [M+H]=535.2 \
N \ N
l , N NI).._\
F
o, 65 NH2 O [M+Hr=527.2 N'/ \N
I ' F
o, 66 NH2 O O [M+H]=510.2 N'' \N
I' N N)___\
\--- )0 F
o, 67 NH2 Ok [M+H]=514.3 ,.N ,..,N
N./\N
I' N )...____\N
----0) F
o, -f-)=1 [M+Hr=514.2 N7 \N N-j 1 IV' N
O
F
0 .
OThr__N
69 NH2 44, Nj_ [M+Hr=514.2 N7 \N
1 ' ---- )0 NH2 4.OTh----\
S/I\I
70 F \ [M+Hr=533.2 N' \N
N
O
F
o, NH2 4.1 -_ 71 _-:--.\.- N [M+H]=516.1 .1µ1 \
N
N N' O
F
F 0 .
72 NH2 0 ,N.,N1 [M+H]=534.2 N \ N
l , N N),.._\
\--- )0 F
0 =
,,õNõN
73 NH2 . [M+Hr=534.1 F
N-- \N
I' N N1).Th Q
F
o, NH2 .
74 --:-.---- A [M+H]=558.2 r.-11 N
I\V \N
I / ----a F
NH2 410 46:1D [M+Hr=518.1 N '7 \
1\1 1 'IV
N
o F
o, NH2 . _--,-L--_ AN [M+Hr=592.1 I\
N V \ ./' N
No 0 F
NH2 ilk 77 -----\ [M+Hr=570.2 N \
I N Or`c',N1 N NI
F
o, 78 NH2 0 8/1=1 1 [M+H]=491.2 N \N
1 ' N N\
7.----F
o, 79 NH2 0 NIN [M+Hr=474.2 r=V \
L I ,N
Th N\
F
0 4, NH2 11*
80 [M+H]=511.2 N 1 \N \1N
N I N' o F
o, OTh--\-1 [M+H]=493.2 N-' \
I'N
N N\._ ----\
OH
F
o, O OTh,...._\
82 NH2 _.
,,N.,1\1 [M+H]=500.2 I ' N N)........, ..---]
F
o, 83 ------\-- [M+H]=560.2 NV \ r N I N
F
o, 0)------N¨ [M+H]=511.2 I
N' \N ' N N).....,.., U
F
o, O\_\
\ [M+H]=485.2 N' \N
I ' N N\_ F
NH2 441i 86 [M+H]=511.2 IµV \N N\/
I NI' O
F
NH2 441) ---_,:--\_. [M+Fir=530.2 N /NI
N \N r N 1 N' o F
o, / \
N
[M+Hr=487.2 N'' \N
1 , N N\_ -----\
OH
F
F [M+H]'=536.1 \N
I' N N)Th Q
_ F
O, 0--\&._ , N
90 NH2 441 N [M+H]'=476.2 N' \N
iN ' Nv ----A
OH
F
O, 91 [M+Hr=543.1 N' \
1_ 1 ,N
i=I N
a F
o, ,.., 92 NvI \N F N" NH2 411 [M+H]=518.2 ' ci F
NH2 *
93 -7---\ [M+H]=502.2 N'' \ H N
r\i/' I N'N
N
o F
O0.
DD / \N
[M+H]'=529.2 N". \
1 N,N
N
O
F
o, NH2 O , \ [M+H]=545.2 N' IN
' N
F
o, / \
N
[M+H]=501.2 N'I \N
, N N\____, \----OH
F
o, NH2 =-,_-__ [M+H]=636.1 rµV " c I N
N
a =
F
98 [M+Hr=492.2 N-- F --N
iµ N' F
o, NH2 fh 99 __--_-\. [M+H]=546.2 NI." \
N I N r--N HO-) O
F
O, SK/i`l \ [M+H]=523.2 NV \
I N ' N
( OH
F
o, 101 H2N *
/ \ N [M+H]=543.1 N' \N
I ' OH
N NI)......_\
F
o, 102 NH2 O N [M+H]4=486.2 -/-"--\
N' IN N'-c ' N>
s-F
o, 0--y,N
103 NH2 * N [M+H]=514.2 N' \
N N),___\
U
F
o, 0-).1..____ 104 NH2 44* N [M+Hr=507.2 N ' \N
I
N' N\_____ \--OH
F
O, 105 NH2 IA ,NN [M+Hr=494.1 F
N' \N
I ' N
---\
OH
_ F
O, 0)__ _ NH N
106 N-----c [M+H]=528.1 N' \N
I ' N Nim '---- )0 F
o, N'-'- [M+H]=512.2 N-' \
L I ,N
N 1`1)_, U
F
o, 108 7----NI_____ [M+H]=530.2 N' \ N
I ,N
N N
o _ F
o, N' Sy/NI
N
I [M+H]=541.3 \
1 ' N 1)____/CI
C
OH
o, 110 NH2 [M+H]=488.1 N
I ' N
OH
fk / \ N
111 NH2 410 [M+H]=515.1 NN
I ' N
HO
o, OTh==.\
112 NH2 40 . [M+H]=504.1 f\V
L ,N
HO
F
0 *
113 NH2 40 SK/ N [M+Hr=505.2 N' \N
I ' N
F
O, ----)--N
114 NH2 *
Nc [M+Hr=502.2 N' "N
I , N r`I.,..,, \--OH
F
0 =
115 NH2 O [M+Hr=490.1 ----______ N N
1 ' N N\_ ---"\
OH
F
, OTh..._:Th_ 116 NH2 Os ,,.-N.,N [M+H]=490.2 N7 \N
I
N ' r`l...._, V_OH
F
0 =
OTh"¨¨\¨
117 NH2 * SK/N1 \ [M+Hr=521.1 N7 \N
I ' HO
F
O, NH2 41i 118 [M+Hr=516.2 L
N1 V 'N HI\J7/N
N NI' F
o, 119 NH2fa [M+Hr=528.2 /-----\N
Ni "N N---;;.1\
N N' F
0 =
120 NH2 41*--1 [M+H]=514.2 N ' \ N
I 'N
N
c-J
F
O, OTh____\
121 NH2 * N1z/ N [M+Hr=518.1 N' I ,N
N N\ i '----/---HO
F
O, 122 NH2 .N'-- [M+H]=516.1 N" \N
I ' HO
F
0 =
123 NH2 * i---Isl...___ [M+H]=488.2 N \ N
I ,N
N N\
/----F
o, 124 NH2 fa '.-----.-I [M+H]=530.1 N' , \
L 1 ,N \
1\1 N
\--70 F
o, 125 NH2 110 ...--..1 _-\- [M+H]=533.2 SK/N
N' \N
1 , \
F
o, 126 NH2 4) [M+H]=527.2 / \ N
N' , \
,N
N N
--i0 F
o, NH2 =127 S.,,i//N [M+Hr=505.2 Nv \N
I ' N 1=1\__ / \---F
0 =
128 NH2 * [M+Hr=501.1 N'i \
I ,N
N N\
F
O, )N-------N
[M+H]=500.1 ---N' \N
I ' F
o, 0---)__ N
130 NH2 * Ni."-c [M+H]=518.1 N' \
1 ,N
N NOH
C
OH
F
o, OTh----\-131 NH2 0 S(/11 \ [M+H]=497.2 NV \N
I' N 1\1\
D3Ct-CD3 F
0 4. DD
0"--\
132 NH2 40 S,,//N [M+H]=493.2 \
N ' \N
I =
F
0 =
133 NH2 Os ft [M+H]=470.1 N-- \
I 'N
N N\
Kinase Binding Btk Kinase Inhibition Assay Fluorescence polarization-based kinase assays were performed in 384 well-plate format using histidine tagged recombinant human full-length Bruton Agammaglobulinemia Tyrosine Kinase (Btk) and a modified protocol of the KinEASE TM FP Fluorescein Green Assay supplied from Millipore. Kinase reaction were performed at room temperature for 60 minutes in presence of 250 DM substrate, 10 OM ATP and variable test article concentrations. The reaction was stopped with EDTA/kinase detection reagents and the polarization measured on a Tecan 500 instrument. From the dose-response curve obtained, the 1050 was calculated using Graph Pad Prisms using a non linear fit curve. The Km for ATP on each enzyme was experimentally determined and the Ki values calculated using the Cheng-Prusoff equation (see: Cheng Y, Prusoff WH. (1973) Relationship between the inhibition constant (K1) and the concentration of inhibitor which causes 50 per cent inhibition (IA of an enzymatic reaction". Biochem Pharmacol 22 (23): 3099-108).
k1 values are reported in Tables 2:
Table 2: Inhibition of Btk Comp 1(1 Comp ki Comp Ici Comp lc, Comp ki ound (nM) ound (nM) ound (nM) ound (nM) ound (nM) 1 a 31 a 61 a 91 a 121 a 2 a 32 a 62 a 92 a 122 a 3 a 33 a 63 a 93 a 123 a 4 a 34 a 64 a 94 - 124 a a 35 a 65 a 95 a 125 a 6 a 36 a 66 a 96 a 126 a 7 a 37 a 67 a 97 a 127 a 8 a 38 a 68 a 98 a 128 a 9 a 39 a 69 a 99 a 129 a a 40 a 70 a 100 a , 130 a 11 a 41 a 71 a 101 a 131 12 a 42 a 72 a 102 a 132 -13 a 43 a 73 a 103 a 133 -14 a 44 a 74 a 104 a a 45 b 75 a 105 a 16 a 46 a 76 a 106 a 17 a 47 a 77 a 107 a 18 a 48 a 78 a 108 a 19 a 49 a 79 a 109 a a 50 a 80 a 110 a 21 a 51 a 81 a 111 a 22 a 52 a 82 a 112 a 23 a 53 a 83 a 113 a 24 a 54 a 84 a 114 a a 55 a 85 a 115 a .
26 a 56 - 86 a 116 a 27 a 57 a 87 a 117 a 28 a 58 a 88 a 118 a 29 a 59 a 89 - 119 a a 60 a 90 a 120 a a - Less than 100 nM; b - less than 1000 nM, c - more than 1000 nM
Splenic Cell Proliferation Assay Splenocytes were obtained from 6 week old male CD1 mice (Charles River Laboratories Inc.). Mouse spleens were manually disrupted in PBS and filtered using a 70um cell strainer followed by ammonium chloride red blood cell lysis. Cells were washed, resuspended in Splenocyte Medium (HyClone RPMI supplemented with 10% heat-inactivated FBS, 0.5X non-essential amino acids, 10mM HEPES, 50uM beta mercaptoethanol) and incubated at 37 C, 5% CO2 for 2h to remove adherent cells. Suspension cells were seeded in 96 well plates at 50,000 cells per well and incubated at 37 C, 5% CO2 for lh.
Splenocytes were pre-treated in triplicate with 10,000 nM curves of Formula 1 compounds for 1h, followed by stimulation of B cell proliferation with 2.5ug/m1 anti-IgM F(ab1)2 (Jackson ImmunoResearch) for 72h. Cell proliferation was measured by Cell Titer-Glo Luminescent Assay (Promega).
EC50 values (50% proliferation in the presence of compound as compared to vehicle treated controls) were calculated from dose response compound curves using GraphPad Prism Software.
EC50 values are reported in Table 3:
Table 3: Inhibition of splenic cell proliferation Compo EC50 Comp EC50 CornEC50 Comp EC50 Comp EC50 poun und (nM) ound (nM) d (nM) ound (nM) ound (nM) 1 b 31 b 61 b 91 a 121 b 2 b 32 a 62 a 92 a 122 b 3 b 33 a 63 a 93 b 123 a 4 b 34 b 64 a 94 a 124 b a 35 a 65 a 95 a 125 a 6 a 36 a 66 b 96 a 126 b 7 b 37 a 67 b 97 b 127 a 8 a 38 a 68 a 98 a 128 a 9 b 39 b 69 a 99 b 129 a b 40 b 70 a 100 a 130 b 11 b 41 b 71 a 101 a 131 12 b 42 a 72 b 102 a 132 -13 a 43 b 73 a 103 a 133 -14 a 44 a 74 b 104 a a 45 b 75 a 105 b 16 b 46 a 76 b 106 a 17 b 47 a 77 a 107 , a 18 a 48 a 78 a 108 a 19 b 49 a 79 a 109 , a a 50 a 80 b 110 b 21 a 51 a 81 a 111 b 22 a 52 a 82 a 112 b 23 a 53 a 83 b 113 a 24 a 54 a 84 a 114 b a 55 a 85 a 115 b 26 a 56 b 86 b 116 a 27 a 57 b 87 a 117 a 28 a 58 a 88 a 118 b 29 a 59 a 89 a 119 b a 60 a 90 b 120 a a - Less than 100 nM; b - less than 1000 nM, c - more than 1000 nM
Methods: Mouse Arthus Mouse Arthus studies were conducted as reported in Braselmann S, Taylor V, Zhao H, Wang S, Sylvain C, Baluom M, Qu K, Herlaar E, Lau A, Young C, Wong BR, Lovell S, Sun T, Park G, Argade A, Jurcevic S, Pine P, Singh R, Grossbard EB, Payan DG, Masuda ES: R406 an orally available spleen tyrosine kinase inhibitor blocks fc receptor signaling and reduces immune-complex mediated inflammation. J Pharmacol Exp Ther, 2006, 319:998-1008.
In summary, female Balb/c mice (6-7 weeks on arrival) were habituated to the animal facility for at least 4 days. On the day of the experiment, animals were pre-treated (t= minus 1 h) with compound or vehicle alone by gavage (PO). At t=0, animals were injected intravenously (IV; 0.1 mL/mouse) with saline containing chicken ovalbumin and Evan's blue (10 mg/mL of each).
Ten minutes later (t= 10 min), animals were anesthesized with isoflurane, the dorsal surface was shaved and rabbit anti-chicken ovalbumin antibody was then injected intradermally at one site on the right side of the animal (25 pg in 30 pL). The same amount of isotype control antibody was then injected on the left side.
The animals were then returned to their home cage and skin punches (8 mm) were collected from each injection site four hours later. The samples were placed in 1 mL formamide overnight at 80 degrees C (1 skin biopsy per 1 mL formamide in a glass tube). The amount of Evan's blue in the formamide solution was then assessed by spectrophotometry (630 nm) as a measure of serum extravasation into the dermis.
Compounds 14, 15, 24, 46, 50, 54, 58, 59, 62, 71, 78, 79, 82, 85, 90, 100, 102, 103, 106, 107, 108, 117, and 125 demonstrated efficacy when administered by oral gavages at 30 mg/kg.
Mouse CIA model was performed using the methods described by Trentham DE, Townes AS, Kang AH. Autoimmunity to Type II Collagen: An Experimental Model of Arthritis. J Exp Med 1977; 857-868, and Bendele AM.
Animal Models of Rheumatoid Arthritis. 3 Musculoskel Interact 2001; 377-385.
In summary, male B1OR111 mice (7-9 wks on arrival) were habituated to the animal facility for at least 4 days. On experimental day 0 mice were anaesthetized with isoflurane and the dorsal surface was shaved. Collagen, emulsified in Freund's complete adjuvant (CFA) supplemented with additional mycobacterium tuberculosis (TB) H37Ra, was injected intradermally at the base of the tail (0.15 mL / animal; 2 mg/mL collagen and 2.5 mg/mL TB in CFA). This CFA treatment was repeated on day 15.
From day 15 to the end of the study animals were scored daily for signs of arthritis. On the first day of disease (RA Day 1) animals were recruited to the study and grouped using a balanced design based on arthritis score. Once recruited, animals were weighed and dosed twice daily by gavage (PO, BID).
Recruited animals were then scored twice a week on RA days 1, 5, 8 and 12.
At the end of the study (RA day 12) animals were weighed and scored.
Compounds 14, 58, 78, 85 and 102 demonstrated efficacy when administered by oral gavages at 30 mg/kg (BID).
Nv_ o, L ,N [M+Hr=543.1 (N--) \--N1 0--)1 NH2 41*
41 N \N [M+Hr=522.1 N' o, 42 N \ N [M+H]=521.2 N#
0¨\
( µj 43 [M+H]=500.2 ,N
N
o, NH2 0--\ 4* US) NI/
44 NI \ [M+H]=530.1 I N
' N Niv -------\
--C) o, 45 NH2 0 40 [M+H]=452.1 NI' \N
I ' N N
/'0 F
o, 46 NH2 44k N [M+H]=519.1 N' \
/ N
' N NI)._.....\
fi ) S
L ,N [M+Fi]=542.1 1%1 /C) fk 48 [M+Hr=498.2 N
,N
N N)_Th 0 =
49 , [M+H]=600.1 L ,N
Thµl r=L
0-<
4Ik , S
50 NH2 =[M+H]=503.2 r%V \
,N
N
o, NH2 Os 51 [M+H]=500.2 N
' N N
L\NH
o, 52 [M+H]=562.1 N'N
I' N N\
\-0 o, OTh---N
53[M+H]=544.2 N'' \N
I' N rµI___ F
0 =
NH2. --==---- \-sN/1µ1 [M+Hr=533.1 /
N- \
l N1 , 1 N N)___\
---'0) F
o, OTh-\--SK./N
[M+H]=576.2 N.-- \N
1' N N___ -\----N3 F
o, 0-Al__ NH2 . N [M+H1=532.2 N' N 1 ' N N
- F
o, NH2 41*
[M+H]=546.2 57 \
NV \N
1 ' N N\
----\
Nc13 F
o, OTh.,__.\
N1 [M+H]=517.2 =
N"
1 ' _.---1 *
NH2 40. SN
[M+Hr=551.1 \N
' N Nym O*
[M+H]=551.2 N" \N
' N N)Th o*
61NH2 O[M+H]=463.2 S,,K/N
N" \
N'N
0 4Ik F
0---\-SK/
NH2 441, N
1 [M+H]=533.2 N' \N
I' N Ny.Th --- )0 F
o, NH2 40 gli [M+H]=512.2 N' \
I N
' N NI)...Th U
F
0 . .V
S/
NH2 41* 1\1 [M+H]=535.2 \
N \ N
l , N NI).._\
F
o, 65 NH2 O [M+Hr=527.2 N'/ \N
I ' F
o, 66 NH2 O O [M+H]=510.2 N'' \N
I' N N)___\
\--- )0 F
o, 67 NH2 Ok [M+H]=514.3 ,.N ,..,N
N./\N
I' N )...____\N
----0) F
o, -f-)=1 [M+Hr=514.2 N7 \N N-j 1 IV' N
O
F
0 .
OThr__N
69 NH2 44, Nj_ [M+Hr=514.2 N7 \N
1 ' ---- )0 NH2 4.OTh----\
S/I\I
70 F \ [M+Hr=533.2 N' \N
N
O
F
o, NH2 4.1 -_ 71 _-:--.\.- N [M+H]=516.1 .1µ1 \
N
N N' O
F
F 0 .
72 NH2 0 ,N.,N1 [M+H]=534.2 N \ N
l , N N),.._\
\--- )0 F
0 =
,,õNõN
73 NH2 . [M+Hr=534.1 F
N-- \N
I' N N1).Th Q
F
o, NH2 .
74 --:-.---- A [M+H]=558.2 r.-11 N
I\V \N
I / ----a F
NH2 410 46:1D [M+Hr=518.1 N '7 \
1\1 1 'IV
N
o F
o, NH2 . _--,-L--_ AN [M+Hr=592.1 I\
N V \ ./' N
No 0 F
NH2 ilk 77 -----\ [M+Hr=570.2 N \
I N Or`c',N1 N NI
F
o, 78 NH2 0 8/1=1 1 [M+H]=491.2 N \N
1 ' N N\
7.----F
o, 79 NH2 0 NIN [M+Hr=474.2 r=V \
L I ,N
Th N\
F
0 4, NH2 11*
80 [M+H]=511.2 N 1 \N \1N
N I N' o F
o, OTh--\-1 [M+H]=493.2 N-' \
I'N
N N\._ ----\
OH
F
o, O OTh,...._\
82 NH2 _.
,,N.,1\1 [M+H]=500.2 I ' N N)........, ..---]
F
o, 83 ------\-- [M+H]=560.2 NV \ r N I N
F
o, 0)------N¨ [M+H]=511.2 I
N' \N ' N N).....,.., U
F
o, O\_\
\ [M+H]=485.2 N' \N
I ' N N\_ F
NH2 441i 86 [M+H]=511.2 IµV \N N\/
I NI' O
F
NH2 441) ---_,:--\_. [M+Fir=530.2 N /NI
N \N r N 1 N' o F
o, / \
N
[M+Hr=487.2 N'' \N
1 , N N\_ -----\
OH
F
F [M+H]'=536.1 \N
I' N N)Th Q
_ F
O, 0--\&._ , N
90 NH2 441 N [M+H]'=476.2 N' \N
iN ' Nv ----A
OH
F
O, 91 [M+Hr=543.1 N' \
1_ 1 ,N
i=I N
a F
o, ,.., 92 NvI \N F N" NH2 411 [M+H]=518.2 ' ci F
NH2 *
93 -7---\ [M+H]=502.2 N'' \ H N
r\i/' I N'N
N
o F
O0.
DD / \N
[M+H]'=529.2 N". \
1 N,N
N
O
F
o, NH2 O , \ [M+H]=545.2 N' IN
' N
F
o, / \
N
[M+H]=501.2 N'I \N
, N N\____, \----OH
F
o, NH2 =-,_-__ [M+H]=636.1 rµV " c I N
N
a =
F
98 [M+Hr=492.2 N-- F --N
iµ N' F
o, NH2 fh 99 __--_-\. [M+H]=546.2 NI." \
N I N r--N HO-) O
F
O, SK/i`l \ [M+H]=523.2 NV \
I N ' N
( OH
F
o, 101 H2N *
/ \ N [M+H]=543.1 N' \N
I ' OH
N NI)......_\
F
o, 102 NH2 O N [M+H]4=486.2 -/-"--\
N' IN N'-c ' N>
s-F
o, 0--y,N
103 NH2 * N [M+H]=514.2 N' \
N N),___\
U
F
o, 0-).1..____ 104 NH2 44* N [M+Hr=507.2 N ' \N
I
N' N\_____ \--OH
F
O, 105 NH2 IA ,NN [M+Hr=494.1 F
N' \N
I ' N
---\
OH
_ F
O, 0)__ _ NH N
106 N-----c [M+H]=528.1 N' \N
I ' N Nim '---- )0 F
o, N'-'- [M+H]=512.2 N-' \
L I ,N
N 1`1)_, U
F
o, 108 7----NI_____ [M+H]=530.2 N' \ N
I ,N
N N
o _ F
o, N' Sy/NI
N
I [M+H]=541.3 \
1 ' N 1)____/CI
C
OH
o, 110 NH2 [M+H]=488.1 N
I ' N
OH
fk / \ N
111 NH2 410 [M+H]=515.1 NN
I ' N
HO
o, OTh==.\
112 NH2 40 . [M+H]=504.1 f\V
L ,N
HO
F
0 *
113 NH2 40 SK/ N [M+Hr=505.2 N' \N
I ' N
F
O, ----)--N
114 NH2 *
Nc [M+Hr=502.2 N' "N
I , N r`I.,..,, \--OH
F
0 =
115 NH2 O [M+Hr=490.1 ----______ N N
1 ' N N\_ ---"\
OH
F
, OTh..._:Th_ 116 NH2 Os ,,.-N.,N [M+H]=490.2 N7 \N
I
N ' r`l...._, V_OH
F
0 =
OTh"¨¨\¨
117 NH2 * SK/N1 \ [M+Hr=521.1 N7 \N
I ' HO
F
O, NH2 41i 118 [M+Hr=516.2 L
N1 V 'N HI\J7/N
N NI' F
o, 119 NH2fa [M+Hr=528.2 /-----\N
Ni "N N---;;.1\
N N' F
0 =
120 NH2 41*--1 [M+H]=514.2 N ' \ N
I 'N
N
c-J
F
O, OTh____\
121 NH2 * N1z/ N [M+Hr=518.1 N' I ,N
N N\ i '----/---HO
F
O, 122 NH2 .N'-- [M+H]=516.1 N" \N
I ' HO
F
0 =
123 NH2 * i---Isl...___ [M+H]=488.2 N \ N
I ,N
N N\
/----F
o, 124 NH2 fa '.-----.-I [M+H]=530.1 N' , \
L 1 ,N \
1\1 N
\--70 F
o, 125 NH2 110 ...--..1 _-\- [M+H]=533.2 SK/N
N' \N
1 , \
F
o, 126 NH2 4) [M+H]=527.2 / \ N
N' , \
,N
N N
--i0 F
o, NH2 =127 S.,,i//N [M+Hr=505.2 Nv \N
I ' N 1=1\__ / \---F
0 =
128 NH2 * [M+Hr=501.1 N'i \
I ,N
N N\
F
O, )N-------N
[M+H]=500.1 ---N' \N
I ' F
o, 0---)__ N
130 NH2 * Ni."-c [M+H]=518.1 N' \
1 ,N
N NOH
C
OH
F
o, OTh----\-131 NH2 0 S(/11 \ [M+H]=497.2 NV \N
I' N 1\1\
D3Ct-CD3 F
0 4. DD
0"--\
132 NH2 40 S,,//N [M+H]=493.2 \
N ' \N
I =
F
0 =
133 NH2 Os ft [M+H]=470.1 N-- \
I 'N
N N\
Kinase Binding Btk Kinase Inhibition Assay Fluorescence polarization-based kinase assays were performed in 384 well-plate format using histidine tagged recombinant human full-length Bruton Agammaglobulinemia Tyrosine Kinase (Btk) and a modified protocol of the KinEASE TM FP Fluorescein Green Assay supplied from Millipore. Kinase reaction were performed at room temperature for 60 minutes in presence of 250 DM substrate, 10 OM ATP and variable test article concentrations. The reaction was stopped with EDTA/kinase detection reagents and the polarization measured on a Tecan 500 instrument. From the dose-response curve obtained, the 1050 was calculated using Graph Pad Prisms using a non linear fit curve. The Km for ATP on each enzyme was experimentally determined and the Ki values calculated using the Cheng-Prusoff equation (see: Cheng Y, Prusoff WH. (1973) Relationship between the inhibition constant (K1) and the concentration of inhibitor which causes 50 per cent inhibition (IA of an enzymatic reaction". Biochem Pharmacol 22 (23): 3099-108).
k1 values are reported in Tables 2:
Table 2: Inhibition of Btk Comp 1(1 Comp ki Comp Ici Comp lc, Comp ki ound (nM) ound (nM) ound (nM) ound (nM) ound (nM) 1 a 31 a 61 a 91 a 121 a 2 a 32 a 62 a 92 a 122 a 3 a 33 a 63 a 93 a 123 a 4 a 34 a 64 a 94 - 124 a a 35 a 65 a 95 a 125 a 6 a 36 a 66 a 96 a 126 a 7 a 37 a 67 a 97 a 127 a 8 a 38 a 68 a 98 a 128 a 9 a 39 a 69 a 99 a 129 a a 40 a 70 a 100 a , 130 a 11 a 41 a 71 a 101 a 131 12 a 42 a 72 a 102 a 132 -13 a 43 a 73 a 103 a 133 -14 a 44 a 74 a 104 a a 45 b 75 a 105 a 16 a 46 a 76 a 106 a 17 a 47 a 77 a 107 a 18 a 48 a 78 a 108 a 19 a 49 a 79 a 109 a a 50 a 80 a 110 a 21 a 51 a 81 a 111 a 22 a 52 a 82 a 112 a 23 a 53 a 83 a 113 a 24 a 54 a 84 a 114 a a 55 a 85 a 115 a .
26 a 56 - 86 a 116 a 27 a 57 a 87 a 117 a 28 a 58 a 88 a 118 a 29 a 59 a 89 - 119 a a 60 a 90 a 120 a a - Less than 100 nM; b - less than 1000 nM, c - more than 1000 nM
Splenic Cell Proliferation Assay Splenocytes were obtained from 6 week old male CD1 mice (Charles River Laboratories Inc.). Mouse spleens were manually disrupted in PBS and filtered using a 70um cell strainer followed by ammonium chloride red blood cell lysis. Cells were washed, resuspended in Splenocyte Medium (HyClone RPMI supplemented with 10% heat-inactivated FBS, 0.5X non-essential amino acids, 10mM HEPES, 50uM beta mercaptoethanol) and incubated at 37 C, 5% CO2 for 2h to remove adherent cells. Suspension cells were seeded in 96 well plates at 50,000 cells per well and incubated at 37 C, 5% CO2 for lh.
Splenocytes were pre-treated in triplicate with 10,000 nM curves of Formula 1 compounds for 1h, followed by stimulation of B cell proliferation with 2.5ug/m1 anti-IgM F(ab1)2 (Jackson ImmunoResearch) for 72h. Cell proliferation was measured by Cell Titer-Glo Luminescent Assay (Promega).
EC50 values (50% proliferation in the presence of compound as compared to vehicle treated controls) were calculated from dose response compound curves using GraphPad Prism Software.
EC50 values are reported in Table 3:
Table 3: Inhibition of splenic cell proliferation Compo EC50 Comp EC50 CornEC50 Comp EC50 Comp EC50 poun und (nM) ound (nM) d (nM) ound (nM) ound (nM) 1 b 31 b 61 b 91 a 121 b 2 b 32 a 62 a 92 a 122 b 3 b 33 a 63 a 93 b 123 a 4 b 34 b 64 a 94 a 124 b a 35 a 65 a 95 a 125 a 6 a 36 a 66 b 96 a 126 b 7 b 37 a 67 b 97 b 127 a 8 a 38 a 68 a 98 a 128 a 9 b 39 b 69 a 99 b 129 a b 40 b 70 a 100 a 130 b 11 b 41 b 71 a 101 a 131 12 b 42 a 72 b 102 a 132 -13 a 43 b 73 a 103 a 133 -14 a 44 a 74 b 104 a a 45 b 75 a 105 b 16 b 46 a 76 b 106 a 17 b 47 a 77 a 107 , a 18 a 48 a 78 a 108 a 19 b 49 a 79 a 109 , a a 50 a 80 b 110 b 21 a 51 a 81 a 111 b 22 a 52 a 82 a 112 b 23 a 53 a 83 b 113 a 24 a 54 a 84 a 114 b a 55 a 85 a 115 b 26 a 56 b 86 b 116 a 27 a 57 b 87 a 117 a 28 a 58 a 88 a 118 b 29 a 59 a 89 a 119 b a 60 a 90 b 120 a a - Less than 100 nM; b - less than 1000 nM, c - more than 1000 nM
Methods: Mouse Arthus Mouse Arthus studies were conducted as reported in Braselmann S, Taylor V, Zhao H, Wang S, Sylvain C, Baluom M, Qu K, Herlaar E, Lau A, Young C, Wong BR, Lovell S, Sun T, Park G, Argade A, Jurcevic S, Pine P, Singh R, Grossbard EB, Payan DG, Masuda ES: R406 an orally available spleen tyrosine kinase inhibitor blocks fc receptor signaling and reduces immune-complex mediated inflammation. J Pharmacol Exp Ther, 2006, 319:998-1008.
In summary, female Balb/c mice (6-7 weeks on arrival) were habituated to the animal facility for at least 4 days. On the day of the experiment, animals were pre-treated (t= minus 1 h) with compound or vehicle alone by gavage (PO). At t=0, animals were injected intravenously (IV; 0.1 mL/mouse) with saline containing chicken ovalbumin and Evan's blue (10 mg/mL of each).
Ten minutes later (t= 10 min), animals were anesthesized with isoflurane, the dorsal surface was shaved and rabbit anti-chicken ovalbumin antibody was then injected intradermally at one site on the right side of the animal (25 pg in 30 pL). The same amount of isotype control antibody was then injected on the left side.
The animals were then returned to their home cage and skin punches (8 mm) were collected from each injection site four hours later. The samples were placed in 1 mL formamide overnight at 80 degrees C (1 skin biopsy per 1 mL formamide in a glass tube). The amount of Evan's blue in the formamide solution was then assessed by spectrophotometry (630 nm) as a measure of serum extravasation into the dermis.
Compounds 14, 15, 24, 46, 50, 54, 58, 59, 62, 71, 78, 79, 82, 85, 90, 100, 102, 103, 106, 107, 108, 117, and 125 demonstrated efficacy when administered by oral gavages at 30 mg/kg.
Mouse CIA model was performed using the methods described by Trentham DE, Townes AS, Kang AH. Autoimmunity to Type II Collagen: An Experimental Model of Arthritis. J Exp Med 1977; 857-868, and Bendele AM.
Animal Models of Rheumatoid Arthritis. 3 Musculoskel Interact 2001; 377-385.
In summary, male B1OR111 mice (7-9 wks on arrival) were habituated to the animal facility for at least 4 days. On experimental day 0 mice were anaesthetized with isoflurane and the dorsal surface was shaved. Collagen, emulsified in Freund's complete adjuvant (CFA) supplemented with additional mycobacterium tuberculosis (TB) H37Ra, was injected intradermally at the base of the tail (0.15 mL / animal; 2 mg/mL collagen and 2.5 mg/mL TB in CFA). This CFA treatment was repeated on day 15.
From day 15 to the end of the study animals were scored daily for signs of arthritis. On the first day of disease (RA Day 1) animals were recruited to the study and grouped using a balanced design based on arthritis score. Once recruited, animals were weighed and dosed twice daily by gavage (PO, BID).
Recruited animals were then scored twice a week on RA days 1, 5, 8 and 12.
At the end of the study (RA day 12) animals were weighed and scored.
Compounds 14, 58, 78, 85 and 102 demonstrated efficacy when administered by oral gavages at 30 mg/kg (BID).
Claims (11)
1. A compound of Formula 1:
wherein R1 is selected from the group consisting of:
1) hydrogen, 2) alkyl, 3) heteroalkyl, 4) carbocyclyl, 5) heterocyclyl, and 6) -C(O)R4;
wherein the alkyl, heteroalkyl, carbocyclyl and heterocyclyl may be further substituted by the groups consisting of:
1) hydroxy, 2) alkoxy, 3) alkyl, 4) -OC(O)R4, 5) -OC(O)NR5R6, 6) -C(O)R4, 7) -C(O)NR5R6, 8) -NR5R6, 9) -NR2C(O)R4, 10) -NR2S(O)n R4 and 11) -NR2C(O)NR5R6;
wherein Y is wherein Z is wherein -YZW is X1 and X2 are independently selected from the group consisting of hydrogen, halogen and cyano;
n is an integer from 0 to 2;
m is an integer from 0 to 2;
m' is an integer from 0 to 2;
W is selected from the group consisting of:
1) alkyl, 2) aralkyl, 3) heteroaralkyl, 4) -OR3, 5) -OC(O)R4, 6) -OC(O)NR5R6, 7) -CH2O-R4, 8) -NR5R6, 9) -NR2C(O)R4, 10) -NR2S(O)n R4 and 11) -NR2C(C)NR5R6;
wherein the alkyl, aralkyl and heteraralkyl may be further substituted;
R2 is hydrogen or alkyl;
R3 is selected from the group consisting of: substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted carbocyclyl, heterocyclyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted aralkyl, and substituted or unsubstituted heteroaralkyl;
R4 is selected from the group consisting of: substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted carbocyclyl, heterocyclyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted aralkyl, and substituted or unsubstituted heteroaralkyl;
R5 and R6 are independently selected from the group consisting of: hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, carbocyclyl, heterocyclyl, aryl and heteroaryl;
alternatively R5 and R6 are fused to form a 3 to 8 membered heterocyclyl ring system.
wherein R1 is selected from the group consisting of:
1) hydrogen, 2) alkyl, 3) heteroalkyl, 4) carbocyclyl, 5) heterocyclyl, and 6) -C(O)R4;
wherein the alkyl, heteroalkyl, carbocyclyl and heterocyclyl may be further substituted by the groups consisting of:
1) hydroxy, 2) alkoxy, 3) alkyl, 4) -OC(O)R4, 5) -OC(O)NR5R6, 6) -C(O)R4, 7) -C(O)NR5R6, 8) -NR5R6, 9) -NR2C(O)R4, 10) -NR2S(O)n R4 and 11) -NR2C(O)NR5R6;
wherein Y is wherein Z is wherein -YZW is X1 and X2 are independently selected from the group consisting of hydrogen, halogen and cyano;
n is an integer from 0 to 2;
m is an integer from 0 to 2;
m' is an integer from 0 to 2;
W is selected from the group consisting of:
1) alkyl, 2) aralkyl, 3) heteroaralkyl, 4) -OR3, 5) -OC(O)R4, 6) -OC(O)NR5R6, 7) -CH2O-R4, 8) -NR5R6, 9) -NR2C(O)R4, 10) -NR2S(O)n R4 and 11) -NR2C(C)NR5R6;
wherein the alkyl, aralkyl and heteraralkyl may be further substituted;
R2 is hydrogen or alkyl;
R3 is selected from the group consisting of: substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted carbocyclyl, heterocyclyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted aralkyl, and substituted or unsubstituted heteroaralkyl;
R4 is selected from the group consisting of: substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted carbocyclyl, heterocyclyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted aralkyl, and substituted or unsubstituted heteroaralkyl;
R5 and R6 are independently selected from the group consisting of: hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, carbocyclyl, heterocyclyl, aryl and heteroaryl;
alternatively R5 and R6 are fused to form a 3 to 8 membered heterocyclyl ring system.
2. The compound of claim 1, wherein W is -OR3, and wherein R3 is selected from the group consisting of: substituted or unsubstituted aralkyl and substituted or unsubstituted heteroaralkyl.
3. The compound of claim 1, wherein W is selected from the group consisting of:
4. The compound according to claim 1, wherein W is selected from the group consisting of:
5. The compound according to any one of claims 1 to 4, wherein R1 is selected from the group consisting of:
6. The compound according to claim 5, wherein R1 is selected from the group consisting of:
7. The compound according to any one of claims 1 to 6, wherein Y is selected from the group consisting of:
8. The compound according to any one of claims 1 to 5, wherein Z is selected from the group consisting of:
9. The compound according to claim 8, wherein Z is:
10. The compound according to any one of claims 1 to 9, wherein -YZW is selected from the group consisting of:
11. A compound selected from the group consisting of:
Priority Applications (13)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CA2813299A CA2813299A1 (en) | 2013-04-17 | 2013-04-17 | Protein kinase inhibitors |
| KR1020147036372A KR101972990B1 (en) | 2012-05-31 | 2013-05-28 | Protein kinase inhibitors |
| KR1020197011438A KR20190043648A (en) | 2012-05-31 | 2013-05-28 | Protein kinase inhibitors |
| CN201380028248.2A CN104379586B (en) | 2012-05-31 | 2013-05-28 | Kinases inhibitor |
| BR112014029718A BR112014029718A2 (en) | 2012-05-31 | 2013-05-28 | protein kinase inhibitors |
| CA2874211A CA2874211A1 (en) | 2012-05-31 | 2013-05-28 | Protein kinase inhibitors |
| EP13796590.1A EP2855484A4 (en) | 2012-05-31 | 2013-05-28 | Protein kinase inhibitors |
| US14/404,497 US9796716B2 (en) | 2012-05-31 | 2013-05-28 | Selective inhibitors of Tec and Src protein kinase families |
| IN2338MUN2014 IN2014MN02338A (en) | 2012-05-31 | 2013-05-28 | |
| JP2015514295A JP6175495B2 (en) | 2012-05-31 | 2013-05-28 | Protein kinase inhibitor |
| PCT/CA2013/000513 WO2013177668A1 (en) | 2012-05-31 | 2013-05-28 | Protein kinase inhibitors |
| RU2014145285A RU2678767C2 (en) | 2012-05-31 | 2013-05-28 | Protein kinase inhibitors |
| HK15109229.7A HK1208460A1 (en) | 2012-05-31 | 2015-09-18 | Protein kinase inhibitors |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CA2813299A CA2813299A1 (en) | 2013-04-17 | 2013-04-17 | Protein kinase inhibitors |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2813299A1 true CA2813299A1 (en) | 2014-10-17 |
Family
ID=51831010
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA2813299A Abandoned CA2813299A1 (en) | 2012-05-31 | 2013-04-17 | Protein kinase inhibitors |
Country Status (1)
| Country | Link |
|---|---|
| CA (1) | CA2813299A1 (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2015077866A1 (en) * | 2013-11-26 | 2015-06-04 | Pharmascience Inc. | Protein kinase inhibitors |
| WO2016187723A1 (en) * | 2015-05-27 | 2016-12-01 | Pharmascience Inc. | Inhibitors of the tec kinase enzyme family |
-
2013
- 2013-04-17 CA CA2813299A patent/CA2813299A1/en not_active Abandoned
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2015077866A1 (en) * | 2013-11-26 | 2015-06-04 | Pharmascience Inc. | Protein kinase inhibitors |
| WO2016187723A1 (en) * | 2015-05-27 | 2016-12-01 | Pharmascience Inc. | Inhibitors of the tec kinase enzyme family |
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