U.S. patent application number 16/141775 was filed with the patent office on 2019-08-29 for combination therapies for treatment of cancer.
The applicant listed for this patent is Araxes Pharma LLC. Invention is credited to Matthew Robert Janes, Liansheng Li, Yi Liu, Matthew Peter Patricelli, Pingda Ren.
Application Number | 20190262342 16/141775 |
Document ID | / |
Family ID | 54207823 |
Filed Date | 2019-08-29 |
View All Diagrams
United States Patent
Application |
20190262342 |
Kind Code |
A1 |
Janes; Matthew Robert ; et
al. |
August 29, 2019 |
COMBINATION THERAPIES FOR TREATMENT OF CANCER
Abstract
Combination therapies for treatment of cancers associated with
mutations in the KRAS gene are provided. Compositions comprising
therapeutic agents for treatment of cancers associated with
mutations in the KRAS gene are also provided.
Inventors: |
Janes; Matthew Robert;
(Encinitas, CA) ; Patricelli; Matthew Peter; (San
Diego, CA) ; Li; Liansheng; (San Diego, CA) ;
Ren; Pingda; (San Diego, CA) ; Liu; Yi; (San
Diego, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Araxes Pharma LLC |
San Diego |
CA |
US |
|
|
Family ID: |
54207823 |
Appl. No.: |
16/141775 |
Filed: |
September 25, 2018 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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14858766 |
Sep 18, 2015 |
10111874 |
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16141775 |
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62195633 |
Jul 22, 2015 |
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62052332 |
Sep 18, 2014 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 31/337 20130101;
A61P 35/02 20180101; A61P 43/00 20180101; A61K 31/517 20130101;
A61K 31/517 20130101; A61P 35/00 20180101; A61K 31/496 20130101;
A61K 31/496 20130101; A61P 1/18 20180101; A61K 31/4745 20130101;
A61P 1/00 20180101; A61K 45/06 20130101; A61K 31/00 20130101; A61P
11/00 20180101; A61K 31/00 20130101; A61K 2300/00 20130101; A61K
2300/00 20130101; A61K 2300/00 20130101 |
International
Class: |
A61K 31/517 20060101
A61K031/517; A61K 45/06 20060101 A61K045/06; A61K 31/4745 20060101
A61K031/4745; A61K 31/00 20060101 A61K031/00; A61K 31/496 20060101
A61K031/496; A61K 31/337 20060101 A61K031/337 |
Claims
1. A method for treating a KRAS, HRAS or NRAS G12C mutant cancer,
the method comprising administering an effective amount of a KRAS,
HRAS or NRAS G12C mutant modulating compound and an additional
therapeutic agent to a subject in need thereof, wherein the KRAS,
HRAS or NRAS G12C mutant modulating compound has the following
structure (I): ##STR00687## or a pharmaceutically acceptable salt,
tautomer, prodrug or stereoisomer thereof, wherein: A is CR.sup.1,
CR.sup.2b, NR.sup.7 or S; B is a bond, CR.sup.1 or CR.sup.2c
G.sup.1 and G.sup.2 are each independently N or CH; W, X and Y are
each independently N, NR.sup.5 or CR.sup.6; Z is a bond, N or
CR.sup.6a or Z is NH when Y is C.dbd.O; L.sup.1 is a bond or
NR.sup.7; L.sup.2 is a bond or alkylene; R.sup.1 is H, cyano, halo,
heterocyclyl, heteroaryl, aryloxy or aryl; R.sup.2a, R.sup.2b and
R.sup.2C are each independently H, halo, hydroxyl, C.sub.1-C.sub.6
alkyl, C.sub.1-C.sub.6 haloalkyl, C.sub.1-C.sub.6 alkoxy,
C.sub.3-C.sub.8 cycloalkyl or aryl; R.sup.3a and R.sup.3b are, at
each occurrence, independently H, --OH, --NH.sub.2, --CO.sub.2H,
halo, cyano, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkynyl,
hydroxylalkly, aminylalkyl, alkylaminylalkyl, cyanoalkyl,
carboxyalkyl, aminocarbonylalkyl or aminocarbonyl; or R.sup.3a and
R.sup.3b join to form a carbocyclic or heterocyclic ring; or
R.sup.3a is H, --OH, --NH.sub.2, --CO.sub.2H, halo, cyano,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkynyl, hydroxylalkly,
aminoalkyl, alkylaminylalkyl, cyanoalkyl, carboxyalkyl,
aminylcarbonylalkyl or aminylcarbonyl, and R.sup.3b joins with
R.sup.4b to form a carbocyclic or heterocyclic ring; R.sup.4a and
R.sup.4b are, at each occurrence, independently H, --OH,
--NH.sub.2, --CO.sub.2H, halo, cyano, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkynyl, hydroxylalkly, aminylalkyl,
alkylaminylalkyl, cyanoalkyl, carboxyalkyl, aminocarbonylalkyl or
aminocarbonyl; or R.sup.4a and R.sup.4b join to form a carbocyclic
or heterocyclic ring; or R.sup.4a is H, --OH, --NH, --CO.sub.2H,
halo, cyano, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkynyl,
hydroxylalkly, aminylalkyl, alkylaminoalkyl, cyanoalkyl,
carboxyalkyl, aminylcarbonylalkyl or aminylcarbonyl, and R.sup.4b
joins with R.sup.3b to form a carbocyclic or heterocyclic ring;
R.sup.5 is, at each occurrence, independently H, C.sub.1-C.sub.6
alkyl or a bond to L.sup.1; R.sup.6 is, at each occurrence,
independently H, oxo, cyano, cyanoalkyl, amino, aminylalkyl,
aminylalkylaminyl, aminocarbonyl, alkylaminyl, haloalkylamino,
hydroxylalkyamino, amindinylalkyl, amidinylalkoxy,
amindinylalkylaminyl, guanidinylalkyl, guanidinylalkoxy,
guanidinylalkylaminyl, C.sub.1-C.sub.6 alkoxy, aminylalkoxy,
alkylcarbonylaminylalkoxy, C.sub.1-C.sub.6 alkyl, heterocyclyl,
heterocyclyloxy, heterocyclylalkyloxy, heterocyclylamino,
heterocyclylalkylamino, heteroaryl, heteroaryloxy,
heteroarylalkyloxy, heteroarylamino, heteroarylalkylamino, aryl,
aryloxy, arylamino, arylalkylamino, arylalkyloxy or a bond to
L.sup.1; R.sup.6a is H, alkyl or a bond to L.sup.1; R.sup.7 is H or
C.sub.1-C.sub.6 alkyl m.sup.1 and m.sup.2 are each independently 1,
2 or 3; indicates a single or double bond such that all valences
are satisfied; and E has the following structure: ##STR00688##
wherein: Q is --C(.dbd.O)--, --C(.dbd.NR.sup.8')--,
--NR.sup.8C(.dbd.O)--, --S(.dbd.O).sub.2-- or
--NR.sup.8S(.dbd.O).sub.2--; R.sup.8 is H, C.sub.1-C.sub.6 alkyl or
hydroxylalkyl; R.sup.8 is H, --OH, --CN or C.sub.1-C.sub.6 alkyl;
and R.sup.9 and R.sup.10 are each independently H, cyano,
C.sub.1-C.sub.6 alkyl, aminylalkyl, alkylaminylalkyl, or
hydroxylalkyl or R.sup.9 and R.sup.10 join to form a carbocyclic or
heterocyclic ring, and wherein at least one of W, X, Y or Z is
CR.sup.6 where R.sup.6 is a bond to L.sup.1 or at least one of W, X
or Y is NR.sup.5, wherein R.sup.5 is a bond to L.sup.1.
2. The method of claim 1, wherein the additional therapeutic agent
is an epidermal growth factor receptor (EGFR) inhibitor,
phosphatidylinositol kinase (PI3K) inhibitor, insulin-like growth
factor receptor (IGF1R) inhibitor, Janus kinase (JAK) inhibitor, a
Met kinase (MET) inhibitor, a SRC family kinase (SFK) inhibitor, a
mitogen-activated protein kinase (MEK) inhibitor, an
extracellular-signal-regulated kinase (ERK) inhibitor, mechanistic
target of rapamycin (mTOR) inhibitor, a topoisomerase inhibitor, a
taxane, an anti-metabolite agent, an alkylating agent or a
taxane.
3. The method of claim 1, wherein the additional therapeutic agent
is an epidermal growth factor receptor (EGFR) inhibitor.
4. The method of claim 3, wherein the epidermal growth factor
receptor (EGFR) inhibitor is Erlotinib, Afatinib or Iressa.
5. The method of claim 1, wherein the additional therapeutic agent
is a phosphatidylinositol-3 kinase (PI3K) inhibitor.
6. The method of claim 5, wherein the phosphatidylinositol kinase
(PI3K) inhibitor is GDC0941, MLN1117, BYL719 (Alpelisib) or BKM120
(Buparlisib).
7. The method of claim 1, wherein the additional therapeutic agent
is an Janus kinase (JAK) inhibitor selected from CYT387, GLPG0634,
Baricitinib, Lestaurtinib, momelotinib, Pacritinib, Ruxolitinib,
and TG101348.
8. The method of claim 1, wherein the additional therapeutic agent
is an MET kinase inhibitor selected from Crizotinib, tivantinib,
AMG337, cabozantinib, foretinib, and neutralizing monoclonal
antibodies to MET such as onartuzumab.
9. The method of claim 1, wherein the additional therapeutic agent
is an insulin-like growth factor receptor (IGF1R) inhibitor.
10. The method of claim 9, wherein the insulin-like growth factor
receptor (IGF1R) inhibitor is NVP-AEW541.
11. The method of claim 1, wherein the additional therapeutic agent
is a non-receptor tyrosine kinase inhibitor.
12. The method of claim 9, wherein the non-receptor tyrosine kinase
inhibitor is an SFK inhibitor.
13. The method of claim 12, wherein the SFK inhibitor is Dasatinib,
Ponatinib, saracatinib, or bosutinib.
14. The method of claim 1, wherein the additional therapeutic agent
is a mitogen-activated protein kinase (MEK) inhibitor.
15. The method of claim 14, wherein the mitogen-activated protein
kinase (MEK) inhibitor is trametinib, selumetinib, cobimetinib,
PD0325901, or RO5126766.
16. The method of claim 1, wherein the additional therapeutic agent
is a protein kinase inhibitor.
17. The method of claim 14, wherein the protein kinase inhibitor is
Afatinib, Axitinib, Bevacizumab, Bostutinib, Cetuximab, Crizotinib,
Dasatinib, Erlotinib, Fostamatinib, Gefitinib, Imatinib, Lapatinib,
Lenvatinib, Ibrutinib, Nilotinib, Panitumumab, Pazopanib,
Pegaptanib, Ranibizumab, Ruxolitinib, Sorafenib, Sunitinib, SU6656,
Trastuzumab, Tofacitinib, Vandetanib or Vemurafenib.
18. The method of claim 1, wherein the additional therapeutic agent
is a topoisomerase inhibitor.
19. The method of claim 18, wherein the topoisomerase inhibitor is
Irinotecan.
20. The method of claim 1, wherein the additional therapeutic agent
is a taxane.
21. The method of claim 20, wherein the taxane is Taxol or
Docetaxel.
22. The method of claim 1, wherein the additional therapeutic agent
is an mTOR inhibitor.
23. The method of claim 22, wherein the mTOR inhibitor is Rapamycin
or MLN0128.
24-32. (canceled)
33. The method of claim 1, wherein the compound is a compound from
Table 1.
34-36. (canceled)
37. The method of claim 1, wherein the cancer is a hematological
cancer, pancreatic cancer, MYH associated polyposis, colorectal
cancer or lung cancer.
38-40. (canceled)
41. A method for inhibiting tumor metastasis in a subject having a
KRAS, HRAS or NRAS G12C mutant cancer, the method comprising
administering an effective amount of a KRAS, HRAS or NRAS G12C
mutant modulating compound and an additional therapeutic agent,
wherein the KRAS, HRAS or NRAS G12C mutant modulating compound has
the following structure (I): ##STR00689## or a pharmaceutically
acceptable salt, tautomer, prodrug or stereoisomer thereof,
wherein: A is CR.sup.1, CR.sup.2b, NR.sup.7 or S; B is a bond,
CR.sup.1 or CR.sup.2c; G.sup.1 and G.sup.2 are each independently N
or CH; W, X and Y are each independently N, NR.sup.5 or CR.sup.6; Z
is a bond, N or CR.sup.6a or Z is NH when Y is C.dbd.O; L.sup.1 is
a bond or NR.sup.7; L.sup.2 is a bond or alkylene; R.sup.1 is H,
cyano, halo, heterocyclyl, heteroaryl, aryloxy or aryl; R.sup.2a,
R.sup.2b and R.sup.2C are each independently H, halo, hydroxyl,
C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 haloalkyl, C.sub.1-C.sub.6
alkoxy, C.sub.3-C.sub.8 cycloalkyl or aryl; R.sup.3a and R.sup.3b
are, at each occurrence, independently H, --OH, --NH.sub.2,
--CO.sub.2H, halo, cyano, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6
alkynyl, hydroxylalkly, aminylalkyl, alkylaminylalkyl, cyanoalkyl,
carboxyalkyl, aminocarbonylalkyl or aminocarbonyl; or R.sup.3a and
R.sup.3b join to form a carbocyclic or heterocyclic ring; or
R.sup.3a is H, --OH, --NH.sub.2, --CO.sub.2H, halo, cyano,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkynyl, hydroxylalkly,
aminoalkyl, alkylaminylalkyl, cyanoalkyl, carboxyalkyl,
aminylcarbonylalkyl or aminylcarbonyl, and R.sup.3b joins with
R.sup.4b to form a carbocyclic or heterocyclic ring; R.sup.4a and
R.sup.4b are, at each occurrence, independently H, --OH,
--NH.sub.2, --CO.sub.2H, halo, cyano, C.sub.1-C.sub.6 alkyl,
C.sub.1-C.sub.6 alkynyl, hydroxylalkly, aminylalkyl,
alkylaminylalkyl, cyanoalkyl, carboxyalkyl, aminocarbonylalkyl or
aminocarbonyl; or R.sup.4a and R.sup.4b join to form a carbocyclic
or heterocyclic ring; or R.sup.4a is H, --OH, --NH.sub.2,
--CO.sub.2H, halo, cyano, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkynyl, hydroxylalkly, aminylalkyl, alkylaminoalkyl, cyanoalkyl,
carboxyalkyl, aminylcarbonylalkyl or aminylcarbonyl, and R.sup.4b
joins with R.sup.3b to form a carbocyclic or heterocyclic ring;
R.sup.5 is, at each occurrence, independently H, C.sub.1-C.sub.6
alkyl or a bond to L.sup.1; R.sup.6 is, at each occurrence,
independently H, oxo, cyano, cyanoalkyl, amino, aminylalkyl,
aminylalkylaminyl, aminocarbonyl, alkylaminyl, haloalkylamino,
hydroxylalkyamino, amindinylalkyl, amidinylalkoxy,
amindinylalkylaminyl, guanidinylalkyl, guanidinylalkoxy,
guanidinylalkylaminyl, C.sub.1-C.sub.6 alkoxy, aminylalkoxy,
alkylcarbonylaminylalkoxy, C.sub.1-C.sub.6 alkyl, heterocyclyl,
heterocyclyloxy, heterocyclylalkyloxy, heterocyclylamino,
heterocyclylalkylamino, heteroaryl, heteroaryloxy,
heteroarylalkyloxy, heteroarylamino, heteroarylalkylamino, aryl,
aryloxy, arylamino, arylalkylamino, arylalkyloxy or a bond to
L.sup.1; R.sup.6a is H, alkyl or a bond to L.sup.1; R.sup.7 is H or
C.sub.1-C.sub.6 alkyl m.sup.1 and m.sup.2 are each independently 1,
2 or 3; indicates a single or double bond such that all valences
are satisfied; and E has the following structure: ##STR00690##
wherein: Q is --C(.dbd.O)--, --C(.dbd.NR.sup.8')--,
--NR.sup.8C(.dbd.O)--, --S(.dbd.O).sub.2-- or
--NR.sup.8S(.dbd.O).sub.2--; R.sup.8 is H, C.sub.1-C.sub.6 alkyl or
hydroxylalkyl; R.sup.8 is H, --OH, --CN or C.sub.1-C.sub.6 alkyl;
and R.sup.9 and R.sup.10 are each independently H, cyano,
C.sub.1-C.sub.6 alkyl, aminylalkyl, alkylaminylalkyl, or
hydroxylalkyl or R.sup.9 and R.sup.10 join to form a carbocyclic or
heterocyclic ring, and wherein at least one of W, X, Y or Z is
CR.sup.6 where R.sup.6 is a bond to L.sup.1 or at least one of W, X
or Y is NR.sup.5, wherein R.sup.5 is a bond to L.sup.1.
42-46. (canceled)
47. A kit comprising a KRAS, HRAS or NRAS G12C mutant modulating
compound, an additional therapeutic agent and directions for use of
the compound and the additional therapeutic agent for treatment of
cancer, wherein the KRAS, HRAS or NRAS G12C mutant modulating
compound has the following structure (I): ##STR00691## or a
pharmaceutically acceptable salt, tautomer, prodrug or stereoisomer
thereof, wherein: A is CR.sup.1, CR.sup.2b, NR.sup.7 or S; B is a
bond, CR.sup.1 or CR.sup.2c G.sup.1 and G.sup.2 are each
independently N or CH; W, X and Y are each independently N,
NR.sup.5 or CR.sup.6; Z is a bond, N or CR.sup.6a or Z is NH when Y
is C.dbd.O; L.sup.1 is a bond or NR.sup.7; L.sup.2 is a bond or
alkylene; R.sup.1 is H, cyano, halo, heterocyclyl, heteroaryl,
aryloxy or aryl; R.sup.2a, R.sup.2b and R.sup.2C are each
independently H, halo, hydroxyl, C.sub.1-C.sub.6 alkyl,
C.sub.1-C.sub.6 haloalkyl, C.sub.1-C.sub.6 alkoxy, C.sub.3-C.sub.8
cycloalkyl or aryl; R.sup.3a and R.sup.3b are, at each occurrence,
independently H, --OH, --NH.sub.2, --CO.sub.2H, halo, cyano,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkynyl, hydroxylalkly,
aminylalkyl, alkylaminylalkyl, cyanoalkyl, carboxyalkyl,
aminocarbonylalkyl or aminocarbonyl; or R.sup.3a and R.sup.3b join
to form a carbocyclic or heterocyclic ring; or R.sup.3a is H, --OH,
--NH.sub.2, --CO.sub.2H, halo, cyano, C.sub.1-C.sub.6 alkyl,
C.sub.2-C.sub.6 alkynyl, hydroxylalkly, aminoalkyl,
alkylaminylalkyl, cyanoalkyl, carboxyalkyl, aminylcarbonylalkyl or
aminylcarbonyl, and R.sup.3b joins with R.sup.4b to form a
carbocyclic or heterocyclic ring; R.sup.4a and R.sup.4b are, at
each occurrence, independently H, --OH, --NH.sub.2, --CO.sub.2H,
halo, cyano, C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkynyl,
hydroxylalkly, aminylalkyl, alkylaminylalkyl, cyanoalkyl,
carboxyalkyl, aminocarbonylalkyl or aminocarbonyl; or R.sup.4a and
R.sup.4b join to form a carbocyclic or heterocyclic ring; or
R.sup.4a is H, --OH, --NH, --CO.sub.2H, halo, cyano,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkynyl, hydroxylalkly,
aminylalkyl, alkylaminoalkyl, cyanoalkyl, carboxyalkyl,
aminylcarbonylalkyl or aminylcarbonyl, and R.sup.4b joins with
R.sup.3b to form a carbocyclic or heterocyclic ring; R.sup.5 is, at
each occurrence, independently H, C.sub.1-C.sub.6 alkyl or a bond
to L.sup.1; R.sup.6 is, at each occurrence, independently H, oxo,
cyano, cyanoalkyl, amino, aminylalkyl, aminylalkylaminyl,
aminocarbonyl, alkylaminyl, haloalkylamino, hydroxylalkyamino,
amindinylalkyl, amidinylalkoxy, amindinylalkylaminyl,
guanidinylalkyl, guanidinylalkoxy, guanidinylalkylaminyl,
C.sub.1-C.sub.6 alkoxy, aminylalkoxy, alkylcarbonylaminylalkoxy,
C.sub.1-C.sub.6 alkyl, heterocyclyl, heterocyclyloxy,
heterocyclylalkyloxy, heterocyclylamino, heterocyclylalkylamino,
heteroaryl, heteroaryloxy, heteroarylalkyloxy, heteroarylamino,
heteroarylalkylamino, aryl, aryloxy, arylamino, arylalkylamino,
arylalkyloxy or a bond to L.sup.1; R.sup.6a is H, alkyl or a bond
to L.sup.1; R.sup.7 is H or C.sub.1-C.sub.6 alkyl m.sup.1 and
m.sup.2 are each independently 1, 2 or 3; indicates a single or
double bond such that all valences are satisfied; and E has the
following structure: ##STR00692## wherein: Q is --C(.dbd.O)--,
--C(.dbd.NR.sup.8')--, --NR.sup.8C(.dbd.O)--, --S(.dbd.O).sub.2--
or --NR.sup.8S(.dbd.O).sub.2--; R.sup.8 is H, C.sub.1-C.sub.6 alkyl
or hydroxylalkyl; R.sup.8 is H, --OH, --CN or C.sub.1-C.sub.6
alkyl; and R.sup.9 and R.sup.10 are each independently H, cyano,
C.sub.1-C.sub.6 alkyl, aminylalkyl, alkylaminylalkyl, or
hydroxylalkyl or R.sup.9 and R.sup.10 join to form a carbocyclic or
heterocyclic ring, and wherein at least one of W, X, Y or Z is
CR.sup.6 where R.sup.6 is a bond to L.sup.1 or at least one of W, X
or Y is NR.sup.5, wherein R.sup.5 is a bond to L.sup.1.
48-49. (canceled)
50. The method of claim 1, wherein R.sup.1 is heteroaryl.
51. The method of claim 1, wherein R.sup.1 is thiophenyl,
pyridinyl, pyridinonyl, pyrimidinyl, benzooxazolyl,
benzoisoxazolyl, benzodioxazolyl, benzoimidazolyl, quinolinyl,
quinolinonyl, dihydroquinolinonyl, tetrahydroquinolinyl,
quinazolinyl, indazolyl, indolinonyl, benzothiophenyl or
dihydrobenzodioxinyl.
52. The method of claim 51, wherein R.sup.1 is substituted with one
or more substituents selected from halo, C.sub.1-C.sub.6 alkyl,
C.sub.1-C.sub.6 alkoxy, or C.sub.2-C.sub.6
alkenylcarbonylaminyl.
53. The method of claim 52, wherein the substituents are selected
from fluoro, chloro, amino and methyl.
54. The method of claim 1, wherein R.sup.1 is indazolyl optionally
substituted with one or more substituents selected from
C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy and halo.
55. The method of claim 1, wherein R.sup.1 has one of the following
structures: ##STR00693## ##STR00694##
56. The method of claim 1, wherein R.sup.2a, R.sup.2b and R.sup.2C
are each independently H or halo.
57. The method of claim 56, wherein halo is chloro or fluoro.
58. The method of claim 1, wherein Q is --C(.dbd.O)--.
59. The method of claim 1, wherein each of R.sup.9 and R.sup.10 are
H.
60. The method of claim 1, wherein E has one of the following
structures: ##STR00695##
61. The method of claim 1, wherein R.sup.3a, R.sup.3b, R.sup.4a and
R.sup.4b are each independently H or C.sub.1-C.sub.6 alkyl.
Description
BACKGROUND
Technical Field
[0001] Embodiments of the present invention are generally directed
to combination therapies for treatment of cancers associated with
mutations in the KRAS gene.
Description of the Related Art
[0002] Ras represents a group of closely related monomeric globular
proteins of 189 amino acids (21 kDa molecular mass) which are
associated with the plasma membrane and which bind either GDP or
GTP. Ras acts as a molecular switch. When Ras contains bound GDP,
it is in the resting or off position and is "inactive." In response
to exposure of the cell to certain growth promoting stimuli, Ras is
induced to exchange its bound GDP for a GTP. With GTP bound, Ras is
"switched on" and is able to interact with and activate other
proteins (its "downstream targets"). The Ras protein itself has a
very low intrinsic ability to hydrolyze GTP back to GDP, thus
turning itself into the off state. Switching Ras off requires
extrinsic proteins termed GTPase-activating proteins (GAPs) that
interact with Ras and greatly accelerate the conversion of GTP to
GDP. Any mutation in Ras which affects its ability to interact with
GAP or to convert GTP back to GDP will result in a prolonged
activation of the protein and consequently a prolonged signal to
the cell telling it to continue to grow and divide. Because these
signals result in cell growth and division, overactive Ras
signaling may ultimately lead to cancer.
[0003] Structurally, Ras proteins contain a G domain which is
responsible for the enzymatic activity of Ras--the guanine
nucleotide binding and the hydrolysis (GTPase reaction). It also
contains a C-terminal extension, known as the CAAX box, which may
be post-translationally modified and is responsible for targeting
the protein to the membrane. The G domain is approximately 21-25
kDa in size and it contains a phosphate binding loop (P-loop). The
P-loop represents the pocket where the nucleotides are bound in the
protein, and this is the rigid part of the domain with conserved
amino acid residues which are essential for nucleotide binding and
hydrolysis (Glycine 12, Threonine 26 and Lysine 16). The G domain
also contains the so called Switch I (residues 30-40) and Switch II
(residues 60-76) regions, both of which are the dynamic parts of
the protein which are often represented as the "spring-loaded"
mechanism because of their ability to switch between the resting
and loaded state. The key interaction is the hydrogen bonds formed
by Threonine-35 and glycine-60 with the .gamma.-phosphate of GTP
which maintain Switch 1 and Switch 2 regions respectively in their
active conformation. After hydrolysis of GTP and release of
phosphate, these two relax into the inactive GDP conformation.
[0004] The most notable members of the Ras subfamily are HRAS, KRAS
and NRAS, mainly for being implicated in many types of cancer.
However, there are many other members including DIRAS1; DIRAS2;
DIRAS3; ERAS; GEM; MRAS; NKIRAS1; NKIRAS2; NRAS; RALA; RALB; RAP1A;
RAP1B; RAP2A; RAP2B; RAP2C; RASD1; RASD2; RASL10A; RASL10B;
RASL11A; RASL11B; RASL12; REM1; REM2; RERG; RERGL; RRAD; RRAS and
RRAS2.
[0005] Mutations in any one of the three main isoforms of RAS
(HRAS, NRAS, or KRAS) genes are among the most common events in
human tumorigenesis. KRAS mutations occur in more than 20% of all
human cancers with the highest levels in pancreatic (.about.90%),
colorectal (.about.40%), and lung (.about.35%), with G12C being a
common mutation (glycine-12 to cysteine). This translates into more
than 150,000 newly diagnosed cases of KRAS driven cancer yearly in
the US alone. These patients have no effective treatment options
and their chances for long term survival are extremely low.
[0006] After many years of failed efforts, the direct targeting of
KRAS was long considered to be impossible. More recently an
approach targeting a specific KRAS mutation, G12C, which accounts
for nearly 50% of KRAS mutant lung cancers, has been reported
(Ostrem et al., Nature 2013, 503:548). We have refined this
strategy to yield quite potent inhibitors of KRAS G12C function in
cells and in vivo. These compounds hold great promise for the
treatment of cancers harboring the KRAS G12C mutation.
[0007] While KRAS is a critical driver mutation in many types of
cancer, its precise role in established tumors is the subject of
some debate. KRAS mutated cancer cells show varied degrees of
growth inhibition when mutant KRAS is depleted, with some lines
showing only modest effects (Singh et al., Cancer Cell 2009,
15:489). Further, even in lines with clear growth dependence on
mutant KRAS, depletion of KRAS does not lead to robust induction of
cell death or apoptosis (Sunaga et al., Mol Cancer Ther 2011,
10:336; Young et al., Cancer Discov 2013, 3:112). Thus, despite the
central role for mutant KRAS in tumorigenesis, it is possible that
inhibition of KRAS alone may not be sufficient for a desirable
clinical outcome.
[0008] Accordingly, while progress has been made in this field,
there remains a need in the art for improved methods for treatment
of KRAS mutant cancers, for example combination therapies. The
present invention fulfills this need and provides further related
advantages.
BRIEF SUMMARY
[0009] In brief, the present invention provides methods for
treatment of cancer, for example cancers associated with mutations
in the KRAS gene. In one embodiment, the disclosure provides a
method for treating a KRAS, HRAS or NRAS G12C mutant cancer, the
method comprising administering an effective amount of a KRAS, HRAS
or NRAS G12C mutant modulating compound and an additional
therapeutic agent to a subject in need thereof. Exemplary cancers
that can be treated by the disclosed method include, but are not
limited to, hematological cancers, pancreatic cancer, MYH
associated polyposis, colorectal cancer and/or lung cancer.
[0010] In a different embodiment, the disclosure provides a method
for inducing apoptosis in a cell population comprising a KRAS, HRAS
or NRAS G12C mutant protein, the method comprising administering an
effective amount of a KRAS, HRAS or NRAS G12C mutant modulating
compound and an additional therapeutic agent.
[0011] In still other embodiments, the disclosure is directed to a
method for inhibiting tumor metastasis in a subject having a KRAS,
HRAS or NRAS G12C mutant cancer, the method comprising
administering an effective amount of a KRAS, HRAS or NRAS G12C
mutant modulating compound and an additional therapeutic agent.
[0012] Pharmaceutical compositions and kits for combination therapy
of different cancers are also provided.
[0013] These and other aspects of the invention will be apparent
upon reference to the following detailed description.
BRIEF DESCRIPTION OF THE DRAWINGS
[0014] In the figures, identical reference numbers identify similar
elements. The sizes and relative positions of elements in the
figures are not necessarily drawn to scale and some of these
elements are arbitrarily enlarged and positioned to improve figure
legibility.
[0015] Further, the particular shapes of the elements as drawn are
not intended to convey any information regarding the actual shape
of the particular elements, and have been solely selected for ease
of recognition in the figures.
[0016] FIG. 1 provides western blots for identifying synergistic
pathways for targeting in combination with KRAS G12C inhibition.
Shown are dot blot arrays for detection of phosphorylated receptor
tyrosine kinases (p-RTK) and phosphorylated signaling kinases (ie.
p-ERK, or p-AKT) for indicated cell lines treated with indicated
K-Ras G12C inhibitor. Notable signaling targets and their
respective pathways that are induced or maintained following
treatment are summarized for various cell lines.
[0017] FIG. 2 depicts western blot analysis of downstream targets
of K-Ras signaling. A) NCI-H358 cells expressing the K-Ras G12C
isoform were treated with a DMSO control or K-Ras G12C inhibitors
II-64, I-153, or I-158 at a concentration of 10 .mu.M (lanes 1-4).
Cells were then treated with DMSO, II-64, I-153, or I-158 in
combination with either the EGFR inhibitor erlotinib at 5 .mu.M
(lanes 5-7) or the PI3K inhibitor GDC0941 at 2 .mu.M (lanes 8-12).
Paclitaxel was used as a positive control. The western blots were
probed with antibodies for p-AKT, p-ERK, total ERK, p-RSK, p-S6 and
cleaved PARP. Cleaved PARP is indicative of apoptosis. B) A549
cells that express the G12S isoform of K-Ras were used a control
for K-Ras G12C inhibitor specificity. A549 cells were treated as in
A). Little to no PARP cleavage was detected.
[0018] FIG. 3 depicts caspase activity in K-Ras G12C cell lines and
control cells lines treated with a K-Ras G12C inhibitor alone,
erlotinib alone, or a combination treatment. Taxol (0.5 .mu.M) was
used as a positive control. Caspase activity was evaluated by
measuring luminescence from cleavable substrate in a caspase assay.
A) K-Ras G12C expressing cell lines NCI-H358, NCI-H2122, and
NCI-H2030 were treated with increasing doses of II-64 (m), II-64+1
.mu.M erlotinib (A), or II-64+5 .mu.M erlotinib (V). B) Control
cell lines NCI-H441, HCT116, A375, and A549 were treated as in A).
C) K-Ras G12C expressing cell lines NCI-H358, NCI-H2122, NCI-H2030,
and NCI-H1792 were treated with increasing doses of II-64 (m) or
II-64+2 .mu.M GDC0941 (A). D) Control cell lines NCI-H441, HCT116,
A375, and A549 were treated as in C).
[0019] FIG. 4 depicts the ability of the compounds disclosed herein
in inhibiting Ras-mediated cell cycle progression and induction of
apoptosis. A) Flow cytometry data demonstrating cell cycle
progression in NCI-H358 cells treated with II-64 alone at 5 .mu.M
or M, II-64+erlotinib (5 .mu.M), or II-64+GDC0941 (2 .mu.M). B) The
average apoptosis response of NCI-H358, NCI-H1792, NCI-H2122, and
SW1573 cell lines generated by flow cytometry as described in
A).
[0020] FIG. 5 depicts apoptosis in NCI-H358 cells treated with
II-64 (10 .mu.M), paclitaxel (1.5 nM), or a combination treatment
(II-64+paclitaxel). The top row shows results from cells that were
pretreated with paclitaxel for 24 hours and then treated with II-64
for an additional 48 hours (72 hours total). The bottom row shows
results from cells that were pretreated with II-64 for 24 hours and
then treated with paclitaxel for an additional 48 hours (72 hours
total). Apoptosis was measured via flow cytometry. Gated
populations=% of subdiploid apoptotic cells.
[0021] FIG. 6 depicts apoptosis in NCI-H358 cells treated with
II-64 (10 .mu.M), docetaxel (1 nM), or a combination treatment
(II-64+docetaxel). The top row shows results from cells that were
pretreated with docetaxel for 24 hours and then treated with II-64
for an additional 48 hours (72 hours total). The bottom row shows
results from cells that were pretreated with II-64 for 24 hours and
then treated with docetaxel for an additional 48 hours (72 hours
total). Apoptosis was measured via flow cytometry. Gated
populations=% of subdiploid apoptotic cells.
[0022] FIG. 7 depicts apoptosis in NCI-H358 cells treated with
II-64 (10 .mu.M), SN38 (5 nM), or a combination treatment
(II-64+SN38). The top row shows results from cells that were
pretreated with SN38 for 24 hours and then treated with II-64 for
an additional 48 hours (72 hours total). The bottom row shows
results from cells that were pretreated with II-64 for 24 hours and
then treated with SN38 for an additional 48 hours (72 hours total).
Apoptosis was measured via flow cytometry. Gated populations=% of
subdiploid apoptotic cells.
[0023] FIG. 8 depicts a tyrosine kinase activity array and caspase
activity in Calu-1 cells. A) An RTK array was used to measure
tyrosine kinase activity in NCI-H358 cells and Calu-1 cells. High
SRC activity was detected in Calu-1 cells. B) Caspase activity was
measured in Calu-1 cells using a Caspase-Glo assay. Cells were
treated for 48 hours with increasing concentrations of I-272 alone,
or I-272+dasatinib (SRC inhibitor; 100 nM), I-272+erlotinib (5
.mu.M), I-272+trametinib (20 nM), or I-272+GDC0941 (1 .mu.M).
I-272+dasatinib induced significantly increased apoptosis.
[0024] FIG. 9 is western blot data for combinations of an exemplary
KRAS G12C inhibitor with an EGFR, MEK or PI3K inhibitor.
[0025] FIG. 10 is another western blot showing data from
combinations of an exemplary G12C inhibitor with PI3KPI3K
inhibition.
[0026] FIG. 11 provides data for combinations of a G12C inhibitor
with an EGFR, EGFR/HER2 or PI3K inhibitor in NCI-H358 cells.
[0027] FIG. 12 is Western blot data from experiments treating
CALU-1 cells with an exemplary G12C inhibitor or Dasunatinib, or
both.
[0028] FIG. 13 presents data for combinations of Dasatinib (Das) or
Sarcatinib (Sarc) with an exemplary G12C inhibitor in various cell
lines.
[0029] FIG. 14 is densitometry data for the gel of FIG. 14.
[0030] FIG. 15 is data for combinations of an exemplary G12C
inhibitor and momelotinib in various cell lines.
[0031] FIG. 16 provides Western blot data for combinations of an
exemplary G12C inhibitor and momelotinib or ruxolitinib in various
cell lines.
DETAILED DESCRIPTION
[0032] In the following description, certain specific details are
set forth in order to provide a thorough understanding of various
embodiments of the invention. However, one skilled in the art will
understand that the invention may be practiced without these
details.
[0033] Unless the context requires otherwise, throughout the
present specification and claims, the word "comprise" and
variations thereof, such as, "comprises" and "comprising" are to be
construed in an open, inclusive sense, that is, as "including, but
not limited to".
[0034] Reference throughout this specification to "one embodiment"
or "an embodiment" means that a particular feature, structure or
characteristic described in connection with the embodiment is
included in at least one embodiment of the present invention. Thus,
the appearances of the phrases "in one embodiment" or "in an
embodiment" in various places throughout this specification are not
necessarily all referring to the same embodiment. Furthermore, the
particular features, structures, or characteristics may be combined
in any suitable manner in one or more embodiments.
[0035] Unless defined otherwise, all technical and scientific terms
used herein have the same meaning as is commonly understood by one
of skill in the art to which this invention belongs. As used in the
specification and claims, the singular form "a," "an," and "the"
include plural references unless the context clearly dictates
otherwise.
[0036] "Amidinyl" refers to a radical of the form
--(C.dbd.NR.sub.a)NR.sub.bR.sub.c, wherein R.sub.a, R.sub.b and
R.sub.c are each independently H or C.sub.1-C.sub.6 alkyl.
[0037] "Amino" refers to the --NH.sub.2 radical.
[0038] "Aminylsulfone" refers to the --S(O).sub.2NH.sub.2
radical.
[0039] "Carboxy" or "carboxyl" refers to the --CO.sub.2H
radical.
[0040] "Cyano" refers to the --CN radical.
[0041] "Guanidinyl" refers to a radical of the form
--NR.sub.d(C.dbd.NR.sub.a)NR.sub.bR.sub.c, wherein R.sub.a,
R.sub.b, R.sub.c and R.sub.d are each independently H or
C.sub.1-C.sub.6 alkyl.
[0042] "Hydroxy" or "hydroxyl" refers to the --OH radical.
[0043] "Imino" refers to the .dbd.NH substituent.
[0044] "Nitro" refers to the --NO.sub.2 radical.
[0045] "Oxo" refers to the .dbd.O substituent.
[0046] "Thioxo" refers to the .dbd.S substituent.
[0047] "Alkyl" refers to a straight or branched hydrocarbon chain
radical consisting solely of carbon and hydrogen atoms, which is
saturated or unsaturated (i.e., contains one or more double and/or
triple bonds), having from one to twelve carbon atoms
(C.sub.1-C.sub.12 alkyl), preferably one to eight carbon atoms
(C.sub.1-C.sub.8 alkyl) or one to six carbon atoms (C.sub.1-C.sub.6
alkyl), and which is attached to the rest of the molecule by a
single bond, e.g., methyl, ethyl, n-propyl, 1-methylethyl
(iso-propyl), n-butyl, n-pentyl, 1,1-dimethylethyl (t-butyl),
3-methylhexyl, 2-methylhexyl, ethenyl, prop-1-enyl, but-1-enyl,
pent-1-enyl, penta-1,4-dienyl, ethynyl, propynyl, butynyl,
pentynyl, hexynyl, and the like. Alkyl includes alkenyls (one or
more carbon-carbon double bonds) and alkynyls (one or more
carbon-carbon triple bonds such as ethynyl and the like).
"Amidinylalkyl" refers to an alkyl group comprising at least one
amidinyl substituent. "Guanidinylalkyl" refers to an alkyl group
comprising at least one guanidinyl substituent. Unless stated
otherwise specifically in the specification, an alkyl,
amidinylalkyl and/or guanidinylalkyl group is optionally
substituted.
[0048] "Alkylene" or "alkylene chain" refers to a straight or
branched divalent hydrocarbon chain linking the rest of the
molecule to a radical group, consisting solely of carbon and
hydrogen, which is saturated or unsaturated (i.e., contains one or
more double and/or triple bonds), and having from one to twelve
carbon atoms, e.g., methylene, ethylene, propylene, n-butylene,
ethenylene, propenylene, n-butenylene, propynylene, n-butynylene,
and the like. The alkylene chain is attached to the rest of the
molecule through a single or double bond and to the radical group
through a single or double bond. The points of attachment of the
alkylene chain to the rest of the molecule and to the radical group
can be through one carbon or any two carbons within the chain.
Unless stated otherwise specifically in the specification, an
alkylene chain is optionally substituted.
[0049] "Alkylcycloalkyl" refers to a radical of the formula
--R.sub.bR.sub.d where R.sub.b is cycloalkyl chain as defined
herein and R.sub.d is an alkyl radical as defined above. Unless
stated otherwise specifically in the specification, an
alkylcycloalkyl group is optionally substituted.
[0050] "Alkoxy" refers to a radical of the formula --OR.sub.a where
R.sub.a is an alkyl radical as defined above containing one to
twelve carbon atoms. "Amidinylalkyloxy" refers to an alkoxy group
comprising at least one amidinyl substituent on the alkyl
group.
[0051] "Guanidinylalkyloxy" refers to an alkoxy group comprising at
least one guanidinyl substituent on the alkyl group.
"Alkylcarbonylaminylalkyloxy" refers to an alkoxy group comprising
at least one alkylcarbonylaminyl substituent on the alkyl
group.
[0052] "Heterocyclylalkyloxy" refers to an alkoxy group comprising
at least one heterocyclyl substituent on the alkyl group.
"Heteroarylalkyloxy" refers to an alkoxy group comprising at least
one heteroaryl substituent on the alkyl group. "Aminylalkyloxy"
refers to an alkoxy group comprising at least one substituent of
the form --NR.sub.aR.sub.b, where R.sub.a and R.sub.b are each
independently H or C.sub.1-C.sub.6 alkyl, on the alkyl group.
Unless stated otherwise specifically in the specification, an
alkoxy, amidinylalkyloxy, guanidinylalkyloxy, alkylcarbonylaminyl,
heterocyclylalkyloxy, heteroarlyalkyloxy and/or aminylalkyloxy
group is optionally substituted.
[0053] "Alkoxyalkyl" refers to a radical of the formula
--R.sub.bOR.sub.a where R.sub.a is an alkyl radical as defined
above containing one to twelve carbon atoms and R.sub.b is an
alkylene radical as defined above containing one to twelve carbon
atoms. Unless stated otherwise specifically in the specification,
an alkoxyalkyl group is optionally substituted.
[0054] "Alkoxycarbonyl" refers to a radical of the formula
--C(.dbd.O)OR.sub.a where R.sub.a is an alkyl radical as defined
above containing one to twelve carbon atoms. Unless stated
otherwise specifically in the specification, an alkoxycarbonyl
group is optionally substituted.
[0055] "Aryloxy" refers to a radical of the formula --OR.sub.a
where R.sub.a is an aryl radical as defined herein. Unless stated
otherwise specifically in the specification, an aryloxy group is
optionally substituted.
[0056] "Alkylaminyl" refers to a radical of the formula --NHR.sub.a
or --NR.sub.aR.sub.a where each R.sub.a is, independently, an alkyl
radical as defined above containing one to twelve carbon atoms. A
"haloalkylaminyl" group is an alkylaminyl group comprising at least
one halo substituent on the alkyl group. A "hydroxylalkylaminyl"
group is an alkylaminyl group comprising at least one hydroxyl
substituent on the alkyl group. An "amidinylalkylaminyl" group is
an alkylaminyl group comprising at least one amidinyl substituent
on the alkyl group. A "guanidinylalkylaminyl" group is an
alkylaminyl group comprising at least one guanidinyl substituent on
the alkyl group. Unless stated otherwise specifically in the
specification, an alkylaminyl, haloalkylaminyl,
hydroxylalkylaminyl, amidinylalkylaminyl and/or
guanidinylalkylaminyl group is optionally substituted.
[0057] "Aminylalkyl" refers to an alkyl group comprising at least
one aminyl substituent (--NR.sub.aR.sub.b wherein R.sub.a and
R.sub.b are each independently H or C.sub.1-C.sub.6 alkyl). The
aminyl substituent can be on a tertiary, secondary or primary
carbon. Unless stated otherwise specifically in the specification,
an aminylalkyl group is optionally substituted.
[0058] "Aminylalkylaminyl" refers to a radical of the formula
--NR.sub.aR.sub.b wherein R.sub.a is H or C.sub.1-C.sub.6 alkyl and
R.sub.b is aminylalkyl. Unless stated otherwise specifically in the
specification, an aminylalkylaminyl group is optionally
substituted.
[0059] "Alkylcarbonylaminyl" refers to a radical of the formula
--NH(C.dbd.O)R.sub.a where R.sub.a is an alkyl radical as defined
above containing one to twelve carbon atoms. Unless stated
otherwise specifically in the specification, an alkylcarbonylaminyl
group is optionally substituted. An alkenylcarbonylaminyl is an
alkylcarbonylaminyl containing at least one carbon-carbon double
bond. An alkenylcarbonylaminyl group is optionally substituted.
[0060] "Alkylaminylalkyl" refers to an alkyl group comprising at
least one alkylaminyl substituent. The alkylaminyl substituent can
be on a tertiary, secondary or primary carbon. Unless stated
otherwise specifically in the specification, an alkylaminylalkyl
group is optionally substituted.
[0061] "Aminylcarbonyl" refers to a radical of the formula
--C(.dbd.O)NH.sub.2. Unless stated otherwise specifically in the
specification, an aminylcarbonyl group is optionally
substituted.
[0062] "Alkylaminylcarbonyl" refers to a radical of the formula
--C(.dbd.O)NR.sub.aR.sub.b, where R.sub.a and R.sub.b are each
independently H or alkyl, provided at least one of R.sub.a or
R.sub.b is alkyl. Unless stated otherwise specifically in the
specification, an alkylaminylcarbonyl group is optionally
substituted.
[0063] "Aminylcarbonylalkyl" refers to a radical of the formula
--R.sub.cC(.dbd.O)NR.sub.aR.sub.b, where R.sub.a and R.sub.b are
each independently H or alkyl and R.sub.c is alkylene. Unless
stated otherwise specifically in the specification, an
aminylcarbonylalkyl group is optionally substituted.
[0064] "Aminylcarbonycycloalkylalkyl" refers to a radical of the
formula --R.sub.cC(.dbd.O)NR.sub.aR.sub.b, where R.sub.a and
R.sub.b are each independently H or alkyl and R.sub.c is
cycloalkyl. Unless stated otherwise specifically in the
specification, an aminylcarbonylcycloalkyl group is optionally
substituted.
[0065] "Aryl" refers to a hydrocarbon ring system radical
comprising hydrogen, 6 to 18 carbon atoms and at least one aromatic
ring. For purposes of this invention, the aryl radical is a
monocyclic, bicyclic, tricyclic or tetracyclic ring system, which
may include fused or bridged ring systems. Aryl radicals include,
but are not limited to, aryl radicals derived from aceanthrylene,
acenaphthylene, acephenanthrylene, anthracene, azulene, benzene,
chrysene, fluoranthene, fluorene, as-indacene, s-indacene, indane,
indene, naphthalene, phenalene, phenanthrene, pleiadene, pyrene,
and triphenylene. Unless stated otherwise specifically in the
specification, the term "aryl" or the prefix "ar-" (such as in
"aralkyl") is meant to include aryl radicals that are optionally
substituted.
[0066] "Aralkyl" refers to a radical of the formula
--R.sub.b--R.sub.e where R.sub.b is an alkylene chain as defined
above and R.sub.c is one or more aryl radicals as defined above,
for example, benzyl, diphenylmethyl and the like. Unless stated
otherwise specifically in the specification, an aralkyl group is
optionally substituted.
[0067] "Carboxyalkyl" refers to a radical of the formula
--R.sub.b--R.sub.e where R.sub.b is an alkylene chain as defined
above and R.sub.c is a carboxy group as defined above. Unless
stated otherwise specifically in the specification, carboxyalkyl
group is optionally substituted.
[0068] "Cyanoalkyl" refers to a radical of the formula
--R.sub.b--R.sub.e where R.sub.b is an alkylene chain as defined
above and R.sub.c is a cyano group as defined above. Unless stated
otherwise specifically in the specification, a cyanoalkyl group is
optionally substituted.
[0069] "Cycloalkyl" or "carbocyclic ring" refers to a stable
non-aromatic monocyclic or polycyclic hydrocarbon radical
consisting solely of carbon and hydrogen atoms, which may include
fused or bridged ring systems, having from three to fifteen carbon
atoms, preferably having from three to ten carbon atoms, and which
is saturated or unsaturated and attached to the rest of the
molecule by a single bond. Monocyclic radicals include, for
example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,
cycloheptyl, and cyclooctyl. Polycyclic radicals include, for
example, adamantyl, norbornyl, decalinyl,
7,7-dimethyl-bicyclo[2.2.1]heptanyl, and the like. A "cycloalkenyl"
is a cycloalkyl comprising one or more carbon-carbon double bonds
within the ring. Unless otherwise stated specifically in the
specification, a cycloalkyl (or cycloalkenyl) group is optionally
substituted.
[0070] "Cyanocycloalkyl" refers to a radical of the formula
--R.sub.b--R.sub.e where R.sub.b is cycloalkylene chain and R.sub.c
is a cyano group as defined above. Unless stated otherwise
specifically in the specification, a cyanocycloalkyl group is
optionally substituted.
[0071] "Cycloalkylaminylcarbonyl" refers to a radical of the
formula --C(.dbd.O)NR.sub.aR.sub.b, where R.sub.a and R.sub.b are
each independently H or cycloalkyl, provided at least one of
R.sub.a or R.sub.b is cycloalkyl. Unless stated otherwise
specifically in the specification, n cycloalkylaminylcarbonyl group
is optionally substituted.
[0072] "Cycloalkylalkyl" refers to a radical of the formula
--R.sub.bR.sub.d where R.sub.b is an alkylene chain as defined
above and R.sub.d is a cycloalkyl radical as defined above. Unless
stated otherwise specifically in the specification, a
cycloalkylalkyl group is optionally substituted.
[0073] "Fused" refers to any ring structure described herein which
is fused to an existing ring structure in the compounds of the
invention. When the fused ring is a heterocyclyl ring or a
heteroaryl ring, any carbon atom on the existing ring structure
which becomes part of the fused heterocyclyl ring or the fused
heteroaryl ring is replaced with a nitrogen atom.
[0074] "Halo" or "halogen" refers to bromo, chloro, fluoro or
iodo.
[0075] "Haloalkyl" refers to an alkyl radical, as defined above,
that is substituted by one or more halo radicals, as defined above,
e.g., trifluoromethyl, difluoromethyl, trichloromethyl,
2,2,2-trifluoroethyl, 1,2-difluoroethyl, 3-bromo-2-fluoropropyl,
1,2-dibromoethyl, and the like. Unless stated otherwise
specifically in the specification, a haloalkyl group is optionally
substituted.
[0076] "Halolkoxy" refers to a radical of the formula --OR.sub.a
where R.sub.a is a haloalkyl radical as defined herein containing
one to twelve carbon atoms. Unless stated otherwise specifically in
the specification, a haloalkoxy group is optionally
substituted.
[0077] "Heterocyclyl" or "heterocyclic ring" refers to a stable 3-
to 18-membered non-aromatic ring radical which consists of two to
twelve carbon atoms and from one to six heteroatoms selected from
the group consisting of nitrogen, oxygen and sulfur. Unless stated
otherwise specifically in the specification, the heterocyclyl
radical is a monocyclic, bicyclic, tricyclic or tetracyclic ring
system, which may include fused or bridged ring systems; and the
nitrogen, carbon or sulfur atoms in the heterocyclyl radical is
optionally oxidized; the nitrogen atom is optionally quaternized;
and the heterocyclyl radical is partially or fully saturated.
Examples of such heterocyclyl radicals include, but are not limited
to, dioxolanyl, thienyl[1,3]dithianyl, decahydroisoquinolyl,
imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl,
morpholinyl, octahydroindolyl, octahydroisoindolyl,
2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl,
oxazolidinyl, piperidinyl, piperazinyl, 4-piperidonyl,
pyrrolidinyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl,
tetrahydrofuryl, trithianyl, tetrahydropyranyl, thiomorpholinyl,
thiamorpholinyl, 1-oxo-thiomorpholinyl, and
1,1-dioxo-thiomorpholinyl. Unless stated otherwise specifically in
the specification. "Heterocyclyloxy" refers to a heterocyclyl group
bound to the remainder of the molecule via an oxygen bond
(--O--).
[0078] "Heterocyclylaminyl" refers to a heterocyclyl group bound to
the remainder of the molecule via a nitrogen bond (--NR.sub.a--,
where R.sub.a is H or C.sub.1-C.sub.6 alkyl). Unless stated
otherwise specifically in the specification, a heterocyclyl,
heterocyclyloxy and/or hetercyclylaminyl group is optionally
substituted.
[0079] "N-heterocyclyl" refers to a heterocyclyl radical as defined
above containing at least one nitrogen and where the point of
attachment of the heterocyclyl radical to the rest of the molecule
is through a nitrogen atom in the heterocyclyl radical. Unless
stated otherwise specifically in the specification, an
N-heterocyclyl group is optionally substituted.
[0080] "Heterocyclylalkyl" refers to a radical of the formula
--R.sub.bR.sub.e where R.sub.b is an alkylene chain as defined
above and R.sub.e is a heterocyclyl radical as defined above, and
if the heterocyclyl is a nitrogen-containing heterocyclyl, the
heterocyclyl is optionally attached to the alkyl radical at the
nitrogen atom. Unless stated otherwise specifically in the
specification, a heterocyclylalkyl group is optionally
substituted.
[0081] "Heteroaryl" refers to a 5- to 14-membered ring system
radical comprising hydrogen atoms, one to thirteen carbon atoms,
one to six heteroatoms selected from the group consisting of
nitrogen, oxygen and sulfur, and at least one aromatic ring. For
purposes of this invention, the heteroaryl radical may be a
monocyclic, bicyclic, tricyclic or tetracyclic ring system, which
may include fused or bridged ring systems; and the nitrogen, carbon
or sulfur atoms in the heteroaryl radical may be optionally
oxidized; the nitrogen atom may be optionally quaternized. Examples
include, but are not limited to, azepinyl, acridinyl,
benzimidazolyl, benzothiazolyl, benzindolyl, benzodioxolyl,
benzofuranyl, benzooxazolyl, benzothiazolyl, benzothiadiazolyl,
benzo[b][1,4]dioxepinyl, 1,4-benzodioxanyl, benzonaphthofuranyl,
benzoxazolyl, benzodioxolyl, benzodioxinyl, benzopyranyl,
benzopyranonyl, benzofuranyl, benzofuranonyl, benzothienyl
(benzothiophenyl), benzotriazolyl,
benzo[4,6]imidazo[1,2-a]pyridinyl, carbazolyl, cinnolinyl,
dibenzofuranyl, dibenzothiophenyl, furanyl, furanonyl,
isothiazolyl, imidazolyl, indazolyl, indolyl, indazolyl,
isoindolyl, indolinyl, isoindolinyl, isoquinolyl, indolizinyl,
isoxazolyl, naphthyridinyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl,
oxiranyl, 1-oxidopyridinyl, 1-oxidopyrimidinyl, 1-oxidopyrazinyl,
1-oxidopyridazinyl, 1-phenyl-1H-pyrrolyl, phenazinyl,
phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl, purinyl,
pyrrolyl, pyrazolyl, pyridinyl, pyrazinyl, pyrimidinyl,
pyridazinyl, quinazolinyl, quinoxalinyl, quinolinyl, quinuclidinyl,
isoquinolinyl, tetrahydroquinolinyl, thiazolyl, thiadiazolyl,
triazolyl, tetrazolyl, triazinyl, and thiophenyl (i.e. thienyl).
"Heteroaryloxy" refers to a heteroaryl group bound to the remainder
of the molecule via an oxygen bond (--O--). "Heteroarylaminyl"
refers to a heteroaryl group bound to the remainder of the molecule
via a nitrogen bond (--NR.sub.a--, where R.sub.a is H or
C.sub.1-C.sub.6 alkyl). Unless stated otherwise specifically in the
specification, a heteroaryl, heteroaryloxy and/or heteroarylaminyl
group is optionally substituted.
[0082] "N-heteroaryl" refers to a heteroaryl radical as defined
above containing at least one nitrogen and where the point of
attachment of the heteroaryl radical to the rest of the molecule is
through a nitrogen atom in the heteroaryl radical. Unless stated
otherwise specifically in the specification, an N-heteroaryl group
is optionally substituted.
[0083] "Heteroarylalkyl" refers to a radical of the formula
--R.sub.bR.sub.f where R.sub.b is an alkylene chain as defined
above and R.sub.f is a heteroaryl radical as defined above. Unless
stated otherwise specifically in the specification, a
heteroarylalkyl group is optionally substituted.
[0084] "Hydroxylalkyl" refers to an alkyl group comprising at least
one hydroxyl substituent. The --OH substituent may be on a primary,
secondary or tertiary carbon. Unless stated otherwise specifically
in the specification, a hydroxylalkyl group is optionally
substituted.
[0085] "Thioalkyl" refers to a radical of the formula --SR.sub.a
where R.sub.a is an alkyl radical as defined above containing one
to twelve carbon atoms. Unless stated otherwise specifically in the
specification, a thioalkyl group is optionally substituted.
[0086] The term "substituted" used herein means any of the above
groups (e.g., alkyl, alkylene, alkylcycloalkyl, alkoxy,
alkoxyalkyl, alkoxycarbonyl, aryloxy, alkylaminyl,
alkylcarbonylaminyl, alkylaminylalkyl, aminylcarbonyl,
alkylaminylcarbonyl, aminylcarbonylalkyl,
aminylcarbonycycloalkylalkyl, thioalkyl, aryl, aralkyl,
carboxyalkyl, cyanoalkyl, cycloalkyl, cyanocycloalkyl,
cycloalkylaminylcarbonyl, cycloalkylalkyl, haloalkyl, haloalkoxy,
heterocyclyl, N-heterocyclyl, heterocyclylalkyl, heteroaryl,
N-heteroaryl and/or heteroarylalkyl) wherein at least one hydrogen
atom is replaced by a bond to a non-hydrogen atoms such as, but not
limited to: a halogen atom such as F, Cl, Br, and I; an oxygen atom
in groups such as hydroxyl groups, alkoxy groups, and ester groups;
a sulfur atom in groups such as thiol groups, thioalkyl groups,
sulfone groups, sulfonyl groups, and sulfoxide groups; a nitrogen
atom in groups such as amines, amides, alkylamines, dialkylamines,
arylamines, alkylarylamines, diarylamines, N-oxides, imides, and
enamines; a silicon atom in groups such as trialkylsilyl groups,
dialkylarylsilyl groups, alkyldiarylsilyl groups, and triarylsilyl
groups; and other heteroatoms in various other groups.
"Substituted" also means any of the above groups in which one or
more hydrogen atoms are replaced by a higher-order bond (e.g., a
double- or triple-bond) to a heteroatom such as oxygen in oxo,
carbonyl, carboxyl, and ester groups; and nitrogen in groups such
as imines, oximes, hydrazones, and nitriles. For example,
"substituted" includes any of the above groups in which one or more
hydrogen atoms are replaced with --NR.sub.gR.sub.h,
--NR.sub.gC(.dbd.O)R.sub.h, --NR.sub.gC(.dbd.O)NR.sub.gR.sub.h,
--NR.sub.gC(.dbd.O)OR.sub.h, --NR.sub.gSO.sub.2R.sub.h,
--OC(.dbd.O)N R.sub.gR.sub.h, --OR.sub.g, --SR.sub.g, --SOR.sub.g,
--SO.sub.2R.sub.g, --OSO.sub.2R.sub.g, --SO.sub.2OR.sub.g,
.dbd.NSO.sub.2R.sub.g, and --SO.sub.2NR.sub.gR.sub.h. "Substituted
also means any of the above groups in which one or more hydrogen
atoms are replaced with --C(.dbd.O)R.sub.g, --C(.dbd.O)OR.sub.g,
--C(.dbd.O)NR.sub.gR.sub.h, --CH.sub.2SO.sub.2R.sub.g,
--CH.sub.2SO.sub.2NR.sub.gR.sub.h. In the foregoing, R.sub.g and
R.sub.h are the same or different and independently hydrogen,
alkyl, alkoxy, alkylaminyl, thioalkyl, aryl, aralkyl, cycloalkyl,
cycloalkylalkyl, haloalkyl, heterocyclyl, N-heterocyclyl,
heterocyclylalkyl, heteroaryl, N-heteroaryl and/or heteroarylalkyl.
"Substituted" further means any of the above groups in which one or
more hydrogen atoms are replaced by a bond to an aminyl, cyano,
hydroxyl, imino, nitro, oxo, thioxo, halo, alkyl, alkoxy,
alkylaminyl, thioalkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl,
haloalkyl, heterocyclyl, N-heterocyclyl, heterocyclylalkyl,
heteroaryl, N-heteroaryl and/or heteroarylalkyl group. In addition,
each of the foregoing substituents may also be optionally
substituted with one or more of the above substituents.
[0087] "Electrophile" or "electrophilic moiety" is any moiety
capable of reacting with a nucleophile (e.g., a moiety having a
lone pair of electrons, a negative charge, a partial negative
charge and/or an excess of electrons, for example a --SH group).
Electrophiles typically are electron poor or comprise atoms which
are electron poor. In certain embodiments an electrophile contains
a positive charge or partial positive charge, has a resonance
structure which contains a positive charge or partial positive
charge or is a moiety in which delocalization or polarization of
electrons results in one or more atom which contains a positive
charge or partial positive charge. In some embodiments, the
electrophiles comprise conjugated double bonds, for example an
.alpha.,.beta.-unsaturated carbonyl or .alpha.,.beta.-unsaturated
thiocarbonyl compound.
[0088] The term "effective amount" or "therapeutically effective
amount" refers to that amount of a compound described herein that
is sufficient to effect the intended application including but not
limited to disease treatment, as defined below. The therapeutically
effective amount may vary depending upon the intended treatment
application (in vivo), or the subject and disease condition being
treated, e.g., the weight and age of the subject, the severity of
the disease condition, the manner of administration and the like,
which can readily be determined by one of ordinary skill in the
art. The term also applies to a dose that will induce a particular
response in target cells, e.g. reduction of platelet adhesion
and/or cell migration. The specific dose will vary depending on the
particular compounds chosen, the dosing regimen to be followed,
whether it is administered in combination with other compounds,
timing of administration, the tissue to which it is administered,
and the physical delivery system in which it is carried.
[0089] As used herein, "treatment" or "treating" refers to an
approach for obtaining beneficial or desired results with respect
to a disease, disorder or medical condition including but not
limited to a therapeutic benefit and/or a prophylactic benefit. By
therapeutic benefit is meant eradication or amelioration of the
underlying disorder being treated. Also, a therapeutic benefit is
achieved with the eradication or amelioration of one or more of the
physiological symptoms associated with the underlying disorder such
that an improvement is observed in the subject, notwithstanding
that the subject may still be afflicted with the underlying
disorder. In certain embodiments, for prophylactic benefit, the
compositions are administered to a subject at risk of developing a
particular disease, or to a subject reporting one or more of the
physiological symptoms of a disease, even though a diagnosis of
this disease may not have been made.
[0090] A "therapeutic effect," as that term is used herein,
encompasses a therapeutic benefit and/or a prophylactic benefit as
described above. A prophylactic effect includes delaying or
eliminating the appearance of a disease or condition, delaying or
eliminating the onset of symptoms of a disease or condition,
slowing, halting, or reversing the progression of a disease or
condition, or any combination thereof.
[0091] The term "co-administration," "administered in combination
with," and their grammatical equivalents, as used herein, encompass
administration of two or more agents to an animal, including
humans, so that both agents and/or their metabolites are present in
the subject at the same time. Co-administration includes
simultaneous administration in separate compositions,
administration at different times in separate compositions, or
administration in a composition in which both agents are
present.
[0092] "Pharmaceutically acceptable salt" includes both acid and
base addition salts.
[0093] "Pharmaceutically acceptable acid addition salt" refers to
those salts which retain the biological effectiveness and
properties of the free bases, which are not biologically or
otherwise undesirable, and which are formed with inorganic acids
such as, but are not limited to, hydrochloric acid, hydrobromic
acid, sulfuric acid, nitric acid, phosphoric acid and the like, and
organic acids such as, but not limited to, acetic acid,
2,2-dichloroacetic acid, adipic acid, alginic acid, ascorbic acid,
aspartic acid, benzenesulfonic acid, benzoic acid,
4-acetamidobenzoic acid, camphoric acid, camphor-10-sulfonic acid,
capric acid, caproic acid, caprylic acid, carbonic acid, cinnamic
acid, citric acid, cyclamic acid, dodecylsulfuric acid,
ethane-1,2-disulfonic acid, ethanesulfonic acid,
2-hydroxyethanesulfonic acid, formic acid, fumaric acid, galactaric
acid, gentisic acid, glucoheptonic acid, gluconic acid, glucuronic
acid, glutamic acid, glutaric acid, 2-oxo-glutaric acid,
glycerophosphoric acid, glycolic acid, hippuric acid, isobutyric
acid, lactic acid, lactobionic acid, lauric acid, maleic acid,
malic acid, malonic acid, mandelic acid, methanesulfonic acid,
mucic acid, naphthalene-1,5-disulfonic acid, naphthalene-2-sulfonic
acid, 1-hydroxy-2-naphthoic acid, nicotinic acid, oleic acid,
orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic
acid, pyroglutamic acid, pyruvic acid, salicylic acid,
4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid,
tartaric acid, thiocyanic acid, p-toluenesulfonic acid,
trifluoroacetic acid, undecylenic acid, and the like.
[0094] "Pharmaceutically acceptable base addition salt" refers to
those salts which retain the biological effectiveness and
properties of the free acids, which are not biologically or
otherwise undesirable. These salts are prepared from addition of an
inorganic base or an organic base to the free acid. Salts derived
from inorganic bases include, but are not limited to, the sodium,
potassium, lithium, ammonium, calcium, magnesium, iron, zinc,
copper, manganese, aluminum salts and the like. Preferred inorganic
salts are the ammonium, sodium, potassium, calcium, and magnesium
salts. Salts derived from organic bases include, but are not
limited to, salts of primary, secondary, and tertiary amines,
substituted amines including naturally occurring substituted
amines, cyclic amines and basic ion exchange resins, such as
ammonia, isopropylamine, trimethylamine, diethylamine,
triethylamine, tripropylamine, diethanolamine, ethanolamine,
deanol, 2-dimethylaminoethanol, 2-diethylaminoethanol,
dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine,
hydrabamine, choline, betaine, benethamine, benzathine,
ethylenediamine, glucosamine, methylglucamine, theobromine,
triethanolamine, tromethamine, purines, piperazine, piperidine,
N-ethylpiperidine, polyamine resins and the like. Particularly
preferred organic bases are isopropylamine, diethylamine,
ethanolamine, trimethylamine, dicyclohexylamine, choline and
caffeine.
[0095] The terms "antagonist" and "inhibitor" are used
interchangeably, and they refer to a compound having the ability to
inhibit a biological function of a target protein, whether by
inhibiting the activity or expression of the protein, such as KRAS,
HRAS or NRAS G12C. Accordingly, the terms "antagonist" and
"inhibitors" are defined in the context of the biological role of
the target protein. While preferred antagonists herein specifically
interact with (e.g. bind to) the target, compounds that inhibit a
biological activity of the target protein by interacting with other
members of the signal transduction pathway of which the target
protein is a member are also specifically included within this
definition. A preferred biological activity inhibited by an
antagonist is associated with the development, growth, or spread of
a tumor.
[0096] The term "agonist" as used herein refers to a compound
having the ability to initiate or enhance a biological function of
a target protein, whether by inhibiting the activity or expression
of the target protein. Accordingly, the term "agonist" is defined
in the context of the biological role of the target polypeptide.
While preferred agonists herein specifically interact with (e.g.
bind to) the target, compounds that initiate or enhance a
biological activity of the target polypeptide by interacting with
other members of the signal transduction pathway of which the
target polypeptide is a member are also specifically included
within this definition.
[0097] As used herein, "agent" or "biologically active agent"
refers to a biological, pharmaceutical, or chemical compound or
other moiety. Non-limiting examples include a simple or complex
organic or inorganic molecule, a peptide, a protein, an
oligonucleotide, an antibody, an antibody derivative, antibody
fragment, a vitamin derivative, a carbohydrate, a toxin, or a
chemotherapeutic compound. Various compounds can be synthesized,
for example, small molecules and oligomers (e.g., oligopeptides and
oligonucleotides), and synthetic organic compounds based on various
core structures. In addition, various natural sources can provide
compounds for screening, such as plant or animal extracts, and the
like.
[0098] "Signal transduction" is a process during which stimulatory
or inhibitory signals are transmitted into and within a cell to
elicit an intracellular response. A modulator of a signal
transduction pathway refers to a compound which modulates the
activity of one or more cellular proteins mapped to the same
specific signal transduction pathway. A modulator may augment
(agonist) or suppress (antagonist) the activity of a signaling
molecule.
[0099] An "anti-cancer agent", "anti-tumor agent" or
"chemotherapeutic agent" refers to any agent useful in the
treatment of a neoplastic condition. One class of anti-cancer
agents comprises chemotherapeutic agents. "Chemotherapy" means the
administration of one or more chemotherapeutic drugs and/or other
agents to a cancer patient by various methods, including
intravenous, oral, intramuscular, intraperitoneal, intravesical,
subcutaneous, transdermal, buccal, or inhalation or in the form of
a suppository.
[0100] The term "cell proliferation" refers to a phenomenon by
which the cell number has changed as a result of division. This
term also encompasses cell growth by which the cell morphology has
changed (e.g., increased in size) consistent with a proliferative
signal.
[0101] The term "selective inhibition" or "selectively inhibit"
refers to a biologically active agent refers to the agent's ability
to preferentially reduce the target signaling activity as compared
to off-target signaling activity, via direct or indirect
interaction with the target.
[0102] "Subject" refers to an animal, such as a mammal, for example
a human. The methods described herein can be useful in both human
therapeutics and veterinary applications. In some embodiments, the
subject is a mammal, and in some embodiments, the subject is
human.
[0103] "Mammal" includes humans and both domestic animals such as
laboratory animals and household pets (e.g., cats, dogs, swine,
cattle, sheep, goats, horses, rabbits), and non-domestic animals
such as wildlife and the like.
[0104] "Radiation therapy" means exposing a subject, using routine
methods and compositions known to the practitioner, to radiation
emitters such as alpha-particle emitting radionuclides (e.g.,
actinium and thorium radionuclides), low linear energy transfer
(LET) radiation emitters (i.e. beta emitters), conversion electron
emitters (e.g. strontium-89 and samarium-153-EDTMP, or high-energy
radiation, including without limitation x-rays, gamma rays, and
neutrons.
[0105] An "anti-cancer agent", "anti-tumor agent" or
"chemotherapeutic agent" refers to any agent useful in the
treatment of a neoplastic condition. One class of anti-cancer
agents comprises chemotherapeutic agents. "Chemotherapy" means the
administration of one or more chemotherapeutic drugs and/or other
agents to a cancer patient by various methods, including
intravenous, oral, intramuscular, intraperitoneal, intravesical,
subcutaneous, transdermal, buccal, or inhalation or in the form of
a suppository.
[0106] "Prodrug" is meant to indicate a compound that may be
converted under physiological conditions or by solvolysis to a
biologically active compound described herein (e.g., compound of
structure (I)). Thus, the term "prodrug" refers to a precursor of a
biologically active compound that is pharmaceutically acceptable.
In some aspects, a prodrug is inactive when administered to a
subject, but is converted in vivo to an active compound, for
example, by hydrolysis. The prodrug compound often offers
advantages of solubility, tissue compatibility or delayed release
in a mammalian organism (see, e.g., Bundgard, H., Design of
Prodrugs (1985), pp. 7-9, 21-24 (Elsevier, Amsterdam). A discussion
of prodrugs is provided in Higuchi, T., et al., "Pro-drugs as Novel
Delivery Systems," A.C.S. Symposium Series, Vol. 14, and in
Bioreversible Carriers in Drug Design, ed. Edward B. Roche,
American Pharmaceutical Association and Pergamon Press, 1987, both
of which are incorporated in full by reference herein. The term
"prodrug" is also meant to include any covalently bonded carriers,
which release the active compound in vivo when such prodrug is
administered to a mammalian subject. Prodrugs of an active
compound, as described herein, are typically prepared by modifying
functional groups present in the active compound in such a way that
the modifications are cleaved, either in routine manipulation or in
vivo, to the parent active compound. Prodrugs include compounds
wherein a hydroxy, amino or mercapto group is bonded to any group
that, when the prodrug of the active compound is administered to a
mammalian subject, cleaves to form a free hydroxy, free amino or
free mercapto group, respectively. Examples of prodrugs include,
but are not limited to, acetate, formate and benzoate derivatives
of a hydroxy functional group, or acetamide, formamide and
benzamide derivatives of an amine functional group in the active
compound and the like.
[0107] The term "in vivo" refers to an event that takes place in a
subject's body.
[0108] The invention disclosed herein is also meant to encompass
all pharmaceutically acceptable compounds of structure (I) being
isotopically-labelled by having one or more atoms replaced by an
atom having a different atomic mass or mass number. Examples of
isotopes that can be incorporated into the disclosed compounds
include isotopes of hydrogen, carbon, nitrogen, oxygen,
phosphorous, fluorine, chlorine, and iodine, such as .sup.2H,
.sup.3H, C, .sup.13C, .sup.14C, .sup.13N, .sup.15N, .sup.15O,
.sup.17O, .sup.18O, .sup.31P, .sup.32P, .sup.35S, .sup.18F,
.sup.36Cl, .sup.123I and .sup.125I, respectively. These
radiolabelled compounds could be useful to help determine or
measure the effectiveness of the compounds, by characterizing, for
example, the site or mode of action, or binding affinity to
pharmacologically important site of action. Certain
isotopically-labelled compounds of structure (I), for example,
those incorporating a radioactive isotope, are useful in drug
and/or substrate tissue distribution studies. The radioactive
isotopes tritium, i.e. .sup.3H, and carbon-14, i.e. .sup.14C, are
particularly useful for this purpose in view of their ease of
incorporation and ready means of detection.
[0109] Substitution with heavier isotopes such as deuterium, i.e.
.sup.2H, may afford certain therapeutic advantages resulting from
greater metabolic stability, for example, increased in vivo
half-life or reduced dosage requirements, and hence are preferred
in some circumstances.
[0110] Substitution with positron emitting isotopes, such as
.sup.11C, .sup.18F, .sup.15O and .sup.13N, can be useful in
Positron Emission Topography (PET) studies for examining substrate
receptor occupancy. Isotopically-labeled compounds of structure (I)
can generally be prepared by conventional techniques known to those
skilled in the art or by processes analogous to those described in
the Preparations and Examples as set out below using an appropriate
isotopically-labeled reagent in place of the non-labeled reagent
previously employed.
[0111] The invention disclosed herein is also meant to encompass
the in vivo metabolic products of the disclosed compounds. Such
products may result from, for example, the oxidation, reduction,
hydrolysis, amidation, esterification, and the like of the
administered compound, primarily due to enzymatic processes.
Accordingly, the invention includes compounds produced by a process
comprising administering a compound of this invention to a mammal
for a period of time sufficient to yield a metabolic product
thereof. Such products are typically identified by administering a
radiolabelled compound of the invention in a detectable dose to an
animal, such as rat, mouse, guinea pig, monkey, or to human,
allowing sufficient time for metabolism to occur, and isolating its
conversion products from the urine, blood or other biological
samples.
[0112] "Stable compound" and "stable structure" are meant to
indicate a compound that is sufficiently robust to survive
isolation to a useful degree of purity from a reaction mixture, and
formulation into an efficacious therapeutic agent.
[0113] Often crystallizations produce a solvate of the compound of
the invention. As used herein, the term "solvate" refers to an
aggregate that comprises one or more molecules of a compound of the
invention with one or more molecules of solvent. In some
embodiments, the solvent is water, in which case the solvate is a
hydrate. Alternatively, in other embodiments, the solvent is an
organic solvent. Thus, the compounds of the present invention may
exist as a hydrate, including a monohydrate, dihydrate,
hemihydrate, sesquihydrate, trihydrate, tetrahydrate and the like,
as well as the corresponding solvated forms. In some aspects, the
compound of the invention is a true solvate, while in other cases,
the compound of the invention merely retains adventitious water or
is a mixture of water plus some adventitious solvent.
[0114] "Optional" or "optionally" means that the subsequently
described event of circumstances may or may not occur, and that the
description includes instances where said event or circumstance
occurs and instances in which it does not. For example, "optionally
substituted aryl" means that the aryl radical may or may not be
substituted and that the description includes both substituted aryl
radicals and aryl radicals having no substitution.
[0115] A "pharmaceutical composition" refers to a formulation of a
compound of the invention and a medium generally accepted in the
art for the delivery of the biologically active compound to
mammals, e.g., humans. Such a medium includes all pharmaceutically
acceptable carriers, diluents or excipients therefor.
[0116] "Pharmaceutically acceptable carrier, diluent or excipient"
includes without limitation any adjuvant, carrier, excipient,
glidant, sweetening agent, diluent, preservative, dye/colorant,
flavor enhancer, surfactant, wetting agent, dispersing agent,
suspending agent, stabilizer, isotonic agent, solvent, or
emulsifier which has been approved by the United States Food and
Drug Administration as being acceptable for use in humans or
domestic animals.
[0117] The compounds of the invention, or their pharmaceutically
acceptable salts may contain one or more asymmetric centers and may
thus give rise to enantiomers, diastereomers, and other
stereoisomeric forms that are defined, in terms of absolute
stereochemistry, as (R)- or (S)- or, as (D)- or (L)- for amino
acids. The present invention is meant to include all such possible
isomers, as well as their racemic and optically pure forms.
Optically active (+) and (-), (R)- and (S)-, or (D)- and (L)-
isomers may be prepared using chiral synthons or chiral reagents,
or resolved using conventional techniques, for example,
chromatography and fractional crystallization. Conventional
techniques for the preparation/isolation of individual enantiomers
include chiral synthesis from a suitable optically pure precursor
or resolution of the racemate (or the racemate of a salt or
derivative) using, for example, chiral high pressure liquid
chromatography (HPLC). When the compounds described herein contain
olefinic double bonds or other centers of geometric asymmetry, and
unless specified otherwise, it is intended that the compounds
include both E and Z geometric isomers. Likewise, all tautomeric
forms are also intended to be included.
[0118] The present invention includes all manner of rotamers and
conformationally restricted states of a compound of the
invention.
[0119] A "stereoisomer" refers to a compound made up of the same
atoms bonded by the same bonds but having different
three-dimensional structures, which are not interchangeable. The
present invention contemplates various stereoisomers and mixtures
thereof and includes "enantiomers", which refers to two
stereoisomers whose molecules are non-superimposable mirror images
of one another.
[0120] A "tautomer" refers to a proton shift from one atom of a
molecule to another atom of the same molecule. The present
invention includes tautomers of any said compounds.
[0121] The chemical naming protocol and structure diagrams used
herein are a modified form of the I.U.P.A.C. nomenclature system,
using the ACD/Name Version 9.07 software program and/or ChemDraw
Ultra Version 11.0.1 software naming program (CambridgeSoft). For
complex chemical names employed herein, a substituent group is
typically named before the group to which it attaches. For example,
cyclopropylethyl comprises an ethyl backbone with a cyclopropyl
substituent. Except as described below, all bonds are identified in
the chemical structure diagrams herein, except for all bonds on
some carbon atoms, which are assumed to be bonded to sufficient
hydrogen atoms to complete the valency.
A. Treatment Methods
[0122] The present disclosure is generally directed to methods for
treatment of various cancers. The present inventors have discovered
that a combination of mutant specific KRAS, NRAS or HRAS G12C
inhibitory molecules with clinically relevant molecular targeted
drugs and/or chemotherapy agents is a surprisingly effective method
for treatment of certain cancers, for example cancers associated
with KRAS, NRAS or HRAS G12C mutant proteins (a "KRAS, HRAS or NRAS
G12C mutant cancer"). In various embodiments, inhibition of mutant
KRAS dramatically sensitizes cancer cells to the described
combination therapies, leading to robust cell death. Such
combination methods have the possibility to greatly improve the
outcomes for patients with tumors harboring the KRAS, NRAS or HRAS
G12C mutation.
[0123] Example 1 describes a strategy for identifying and assessing
potential targets that would benefit from a combination treatment
including a KRAS G12C inhibitor and a second agent that would
inhibit any cell signaling pathway that was hyperactivated or
maintained following KRAS G12C inhibition. Exemplary targets
identified include RTK, PI3K, mTOR, SRC, and JAK/STAT.
[0124] Examples 2-10 describe additional data obtained in support
of certain embodiments of the present invention. In Example 2, a
combination of an exemplary KRAS G12C inhibitor was used with one
of an RTK, a PI3K, an mTOR, an SRC, or a JAK inhibitor. The
synergistic effect of the combination was assessed by monitoring
both cell proliferation and apoptosis in the presence of each agent
alone and in the presence of the combination. This example was
carried out on a variety of mutant cell lines (H358, H1792, Calu-1,
SW1463, SW1573, MiaPaca2, NCI-H23) or control cell line (A549). The
proliferation data was combined with apoptosis data to assess the
synergistic effect of the compounds used in combination.
[0125] In Example 3, multiple KRAS G12C cell lines were evaluated
for evidence of apoptosis induction in the presence of KRAS G12C
inhibitors alone, or in combination with targeted agents (EGFR,
PI3K, IGF1R, and MEK inhibitors) or chemotherapeutic agents (Taxol,
Docetaxel, SN38 (active ingredient in Irinotecan)). In similar
fashion, Examples 4-10 each show an evaluation of a KRAS G12C
inhibitor used in combination with one of many different types of
inhibitors (e.g., EGFR, PI3K, MEK, SRC, JAK) for synergistic
pathway inhibition or induction of apoptosis in relevant cancer
cell lines.
[0126] Synergistic apoptosis induction was observed with treatment
of KRAS G12C mutated cell lines with combinations of a KRAS G12C
inhibitor and selected targeted and chemotherapeutic agents. In
many cases, the levels of apoptosis induced by these combinations
rivaled what is seen with high dose staurosporine or Taxol. As with
the single agent treatments, the effects varied by cell-type and by
the precise combination. The greatest synergy was observed when a
KRAS G12C inhibitor was combined with an EGFR inhibitor (e.g.,
erlotinib and afatinib), a PI3K inhibitor (e.g., GDC0941, BYL-719),
or the chemotherapeutic agent SN38 (active ingredient in
Irinotecan). Some positive results were also seen with MEK
inhibitors, IGF1R inhibitors, JAK inhibitors, SRC inhibitors, and
the chemotherapeutic agents Taxol, Docetaxel, and Paclitaxel.
[0127] Overall the data support that inhibition of mutant KRAS
activity can dramatically alter the sensitivity of KRAS mutant cell
lines to both targeted cancer therapies and, more broadly, active
chemotherapies. The levels of apoptosis observed with optimal
combinations are equal to potent apoptosis inducers and suggest
that nearly 100% cell killing is possible. In a clinical setting
this type of behavior should allow for optimal therapeutic
combinations to lead to dramatic tumor regression.
[0128] The effectiveness of different combinations varied with the
different cell lines despite sharing the KRAS G12C mutation. While
not wishing to be bound by theory, this is may be related to
differences in the genetic background and varied oncogene
addictions in each of these lines. FIG. 9 provides data for an
effective combination strategy for Calu-1 cells. Calu-1 cells were
generally resistant to single agent KRAS G12C treatment as well as
combinations with targeted kinase inhibitors. Comparison of
tyrosine kinase phosphorylation levels between H358 and Calu-1
cells revealed that Calu-1 cells have high levels of c-SRC
phosphorylation (FIG. 9A). Combination treatment of Calu-1 cells
with a KRAS G12C inhibitor and a SRC inhibitor (Dasatinib) revealed
a strong synergistic induction of apoptosis (FIG. 9B).
[0129] Given that KRAS is a centrally important oncogene that
universally leads to treatment-resistant cancer, we anticipate that
eliminating the oncogenic KRAS signaling will uncover enhanced
sensitivities to a broad range of cancer therapeutics beyond what
has been examined here.
[0130] Accordingly, in one embodiment a method for treating a KRAS,
HRAS or NRAS G12C mutant cancer is provided, the method comprising
administering an effective amount of a KRAS, HRAS or NRAS G12C
mutant modulating compound and an additional therapeutic agent to a
subject in need thereof. In certain embodiments, the cancer is a
KRAS G12C mutant cancer. The KRAS, HRAS or NRAS G12C mutant
modulating compound is not particularly limited provided the
compound modulates (e.g., inhibits) the activity of the KRAS, HRAS
or NRAS G12C mutant. Exemplary compounds for this purpose are
described herein in the section entitled "Compounds." In various
embodiments of the method, the additional therapeutic agent is an
epidermal growth factor receptor (EGFR) inhibitor,
phosphatidylinositol kinase (PI3K) inhibitor, insulin-like growth
factor receptor (IGF1R) inhibitor, Janus kinase (JAK) inhibitor, a
Met (MET) kinase inhibitor, a SRC family kinase (SFK) inhibitor, a
mitogen-activated protein kinase (MEK) inhibitor, an
extracellular-signal-regulated kinase (ERK) inhibitor, mechanistic
target of rapamycin (mTOR) inhibitor, a topoisomerase inhibitor
(such as irinotecan, or such as etoposide, or such as doxorubicin),
taxanes (such as anti-microtubule agents including paclitaxel and
docetaxel), anti-metabolite agents (such as 5-FU or such as
gemcitabine), alkylating agents (such as cisplatin or such as
cyclophosphamide), or a taxane.
[0131] In some other embodiments of the method, the additional
therapeutic agent is an epidermal growth factor receptor (EGFR)
inhibitor, phosphatidylinositol kinase (PI3K) inhibitor,
insulin-like growth factor receptor (IGF1R) inhibitor, Janus kinase
(JAK) inhibitor, a Met (MET) kinase inhibitor, a SRC family kinase
(SFK) inhibitor, a mitogen-activated protein kinase (MEK)
inhibitor, an extracellular-signal-regulated kinase (ERK)
inhibitor, a topoisomerase inhibitor (such as irinotecan, or such
as etoposide, or such as doxorubicin), taxanes (such as
anti-microtubule agents including paclitaxel and docetaxel),
anti-metabolite agents (such as 5-FU or such as gemcitabine),
alkylating agents (such as cisplatin or such as cyclophosphamide),
or a taxane.
[0132] In some embodiments, the additional therapeutic agent is an
epidermal growth factor receptor (EGFR) inhibitor, such as
erlotinib or such as afatinib. In some embodiments the additional
therapeutic agent is Iressa. In some embodiments the additional
therapeutic agent is a monoclonal antibody such as cetuximab
(Erbitux) or panitumumab (Vectibix). In some embodiments the GFR
inhibitor is a dual or pan-HER inhibitor. In other embodiments, the
additional therapeutic agent is a phosphatidylinositol-3 kinase
(PI3K) inhibitor, such as GDC0941, MLN1117, BYL719 (Alpelisib) or
BKM120 (Buparlisib). GDC0941 refers to
2-(1H-indazol-4-yl)-6-(4-methanesulfonyl-piperazin-1-ylmethyl)-4-morpholi-
n-4-yl-thieno[3,2-d]pyrimidine or a salt thereof (e.g., bismesylate
salt).
[0133] In still different embodiments, the additional therapeutic
agent is an insulin-like growth factor receptor (IGF1R) inhibitor.
For example, in some embodiments the insulin-like growth factor
receptor (IGF1R) inhibitor is NVP-AEW541. In other embodiments, the
additional therapeutic agent is IGOSI-906 (Linsitinib), BMS-754807,
or in other embodiments the additional therapeutic agent is a
neutralizing monoclonal antibodies specific to IGF1R such as
AMG-479 (ganitumab), CP-751,871 (figitumumab), IMC-A12
(cixutumumab), MK-0646 (dalotuzumab), and R-1507 (robatumumab).
[0134] In some other embodiments, the additional therapeutic agent
is a Janus kinase (JAK) inhibitor. In some embodiments, the
additional therapeutic agent is CYT387, GLPG0634, Baricitinib,
Lestaurtinib, momelotinib, Pacritinib, Ruxolitinib or TG101348 In
some other embodiments the additional therapeutic agent is an MET
kinase inhibitor, such as Crizotinib, tivantinib, AMG337,
cabozantinib, foretinib. In other embodiments the additional
therapeutic agent is a neutralizing monoclonal antibody to MET such
as onartuzumab.
[0135] In more embodiments, the additional therapeutic agent is a
SRC family non-receptor tyrosine kinase inhibitor. For example in
some embodiments the additional therapeutic agent is an inhibitor
of the subfamily of SRC family non-receptor tyrosine kinases.
Exemplary inhibitors in this respect include Dasatinib. Other
examples in this regard include Ponatinib, sarcatinib, and
bosutinib In yet different embodiments, the additional therapeutic
agent is a mitogen-activated protein kinase (MEK) inhibitor. In
some of these embodiments, the mitogen-activated protein kinase
(MEK) inhibitor is trametinib, selumetinib, cobimetinib, PD0325901,
or RO5126766. In other embodiments the MEK inhibitor is
GSK-1120212, also known as trametinib.
[0136] In yet different embodiments, the additional therapeutic
agent is an extracellular-signal-regulated kinase (ERK) inhibitor.
In some of these embodiments, the extracellular-signal-regulated
kinase (ERK) inhibitor is SCH722984 or GDC-0994.
[0137] In other embodiments, the additional therapeutic agent is a
protein kinase inhibitor, such as Staurosporine or Midostaurin. In
other embodiments the protein kinase inhibitor is isAfatinib,
Axitinib, Bevacizumab, Bostutinib, Cetuximab, Crizotinib,
Dasatinib, Erlotinib, Fostamatinib, Gefitinib, Imatinib, Lapatinib,
Lenvatinib, Ibrutinib, Nilotinib, Panitumumab, Pazopanib,
Pegaptanib, Ranibizumab, Ruxolitinib, Sorafenib, Sunitinib, SU6656,
Trastuzumab, Tofacitinib, Vandetanib, or Vemurafenib. In still more
embodiments, the additional therapeutic agent is a topoisomerase
inhibitor. In some of these embodiments, the topoisomerase
inhibitor is Irinotecan. In some more embodiments, the additional
therapeutic agent is a taxane. Exemplary taxanes include Taxol and
Docetaxel.
[0138] In still different embodiments, wherein the additional
therapeutic agent is an mTOR inhibitor, such as Rapamycin or
MLN0128.
[0139] The exact method for administering the compound and the
additional therapeutic agent will be apparent to one of ordinary
skill in the art. In some exemplary embodiments the compound and
the additional therapeutic agent are co-administered. In other
embodiments, the compound and the additional therapeutic agent are
separately administered.
[0140] In some embodiments, the compound and the additional
therapeutic agent are administered with the second agent
simultaneously or separately. This administration in combination
can include simultaneous administration of the two agents in the
same dosage form, simultaneous administration in separate dosage
forms, and separate administration. That is, the compound and any
of the additional therapeutic agents described herein can be
formulated together in the same dosage form and administered
simultaneously. Alternatively, the compound and any of the
additional therapeutic agents described herein can be
simultaneously administered, wherein both the agents are present in
separate formulations. In another alternative, the compound can be
administered just followed by and any of the additional therapeutic
agents described herein, or vice versa. In some embodiments of the
separate administration protocol, the compound and any of the
additional therapeutic agents described herein are administered a
few minutes apart, or a few hours apart, or a few days apart.
[0141] While not wishing to be bound by theory, it is believed that
the effectiveness of the presently described combination therapies
is related, at least in part, to the synergistic ability of the
individual components to induce apoptosis in cells comprising KRAS,
HRAS or NRAS G12C mutant protein. Accordingly, in certain
embodiments a method for inducing apoptosis in a cell population
comprising a KRAS, HRAS or NRAS G12C mutant protein is provided,
the method comprising administering an effective amount of a KRAS,
HRAS or NRAS G12C mutant modulating compound and an additional
therapeutic agent. Compounds and additional therapeutic agents
useful in such methods include any of these described herein.
[0142] The described methods also find utility for inhibiting tumor
metastasis, and in some embodiments, a method for inhibiting tumor
metastasis in a subject having a KRAS, HRAS or NRAS G12C mutant
cancer is provided, the method comprising administering an
effective amount of a KRAS, HRAS or NRAS G12C mutant modulating
compound and an additional therapeutic agent. Compounds and
additional therapeutic agents useful for inhibiting tumor
metastasis include any of these described herein.
[0143] The described methods are generally applicable to any type
of cancer. In certain embodiments the cancer is associated with a
KRAS, HRAS or NRAS G12C mutant protein. In some more specific
embodiments, the cancer is associated with a KRAS, HRAS or NRAS
G12C mutant protein. While many cancers are can be treated
according to the disclosed methods, some embodiments are directed
to treatment of hematological cancer, pancreatic cancer, MYH
associated polyposis, colorectal cancer or lung cancer.
[0144] KRAS, HRAS or NRAS G12C mutations have also been identified
in hematological malignancies (e.g., cancers that affect blood,
bone marrow and/or lymph nodes). Accordingly, certain embodiments
of the methods are directed to treatment of a hematological
malignancy. Such malignancies include, but are not limited to
leukemias and lymphomas. For example, the presently disclosed
combination therapy can be used for treatment of diseases such as
Acute lymphoblastic leukemia (ALL), Acute myelogenous leukemia
(AML), Chronic lymphocytic leukemia (CLL), small lymphocytic
lymphoma (SLL), Chronic myelogenous leukemia (CML), Acute monocytic
leukemia (AMoL) and/or other leukemias. In other embodiments, the
methods are useful for treatment of lymphomas such as all subtypes
of Hodgkin's lymphoma or non-Hodgkin's lymphoma.
[0145] Determining whether a tumor or cancer comprises a G12C KRAS,
HRAS or NRAS mutation can be undertaken by assessing the nucleotide
sequence encoding the KRAS, HRAS or NRAS protein, by assessing the
amino acid sequence of the KRAS, HRAS or NRAS protein, or by
assessing the characteristics of a putative KRAS, HRAS or NRAS
mutant protein. The sequence of wild-type human KRAS, HRAS or NRAS
is known in the art, (e.g., Accession No. NP203524).
[0146] Methods for detecting a mutation in a KRAS, HRAS or NRAS
nucleotide sequence are known by those of skill in the art. These
methods include, but are not limited to, polymerase chain
reaction-restriction fragment length polymorphism (PCR-RFLP)
assays, polymerase chain reaction-single strand conformation
polymorphism (PCR-SSCP) assays, real-time PCR assays, PCR
sequencing, mutant allele-specific PCR amplification (MASA) assays,
direct sequencing, primer extension reactions, electrophoresis,
oligonucleotide ligation assays, hybridization assays, TaqMan
assays, SNP genotyping assays, high resolution melting assays and
microarray analyses. In some embodiments, samples are evaluated for
G12C KRAS, HRAS or NRAS mutations by real-time PCR. In real-time
PCR, fluorescent probes, specific for the KRAS, HRAS or NRAS G12C
mutation, are used. When a mutation is present, the probe binds and
fluorescence is detected. In some embodiments, the KRAS, HRAS or
NRAS G12C mutation is identified using a direct sequencing method
of specific regions (e.g., exon 2 and/or exon 3) in the KRAS, HRAS
or NRAS gene. This technique will identify all possible mutations
in the region sequenced.
[0147] Methods for detecting a mutation in a KRAS, HRAS or NRAS
protein are known by those of skill in the art. These methods
include, but are not limited to, detection of a KRAS, HRAS or NRAS
mutant using a binding agent (e.g., an antibody) specific for the
mutant protein, protein electrophoresis and Western blotting, and
direct peptide sequencing.
[0148] Methods for determining whether a tumor or cancer comprises
a G12C KRAS, HRAS or NRAS mutation can use a variety of samples. In
some embodiments, the sample is taken from a subject having a tumor
or cancer. In some embodiments, the sample is taken from a subject
having a cancer or tumor. In some embodiments, the sample is a
fresh tumor/cancer sample. In some embodiments, the sample is a
frozen tumor/cancer sample. In some embodiments, the sample is a
formalin-fixed paraffin-embedded sample. In some embodiments, the
sample is processed to a cell lysate. In some embodiments, the
sample is processed to DNA or RNA.
[0149] In some embodiments, the disclosed methods are for treating
a hyperproliferative disorder in a subject that comprises
administering to said subject a therapeutically effective amount of
a KRAS, HRAS or NRAS G12C mutant modulating compound and an
additional therapeutic agent to a subject in need thereof. The
compounds and additional therapeutic reagents useful in this regard
include any of those described herein. In some embodiments, said
method relates to the treatment of cancer such as acute myeloid
leukemia, cancer in adolescents, adrenocortical carcinoma
childhood, AIDS-related cancers (e.g. Lymphoma and Kaposi's
Sarcoma), anal cancer, appendix cancer, astrocytomas, atypical
teratoid, basal cell carcinoma, bile duct cancer, bladder cancer,
bone cancer, brain stem glioma, brain tumor, breast cancer,
bronchial tumors, burkitt lymphoma, carcinoid tumor, atypical
teratoid, embryonal tumors, germ cell tumor, primary lymphoma,
cervical cancer, childhood cancers, chordoma, cardiac tumors,
chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia
(CML), chronic myleoproliferative disorders, colon cancer,
colorectal cancer, craniopharyngioma, cutaneous T-cell lymphoma,
extrahepatic ductal carcinoma in situ (DCIS), embryonal tumors, CNS
cancer, endometrial cancer, ependymoma, esophageal cancer,
esthesioneuroblastoma, ewing sarcoma, extracranial germ cell tumor,
extragonadal germ cell tumor, eye cancer, fibrous histiocytoma of
bone, gall bladder cancer, gastric cancer, gastrointestinal
carcinoid tumor, gastrointestinal stromal tumors (GIST), germ cell
tumor, gestational trophoblastic tumor, hairy cell leukemia, head
and neck cancer, heart cancer, liver cancer, Hodgkin's lymphoma,
hypopharyngeal cancer, intraocular melanoma, islet cell tumors,
pancreatic neuroendocrine tumors, kidney cancer, laryngeal cancer,
lip and oral cavity cancer, liver cancer, lobular carcinoma in situ
(LCIS), lung cancer, lymphoma, metastatic squamous neck cancer with
occult primary, midline tract carcinoma, mouth cancer multiple
endocrine neoplasia syndromes, multiple myeloma/plasma cell
neoplasm, mycosis fungoides, myelodysplastic syndromes,
myelodysplastic/myleoproliferative neoplasms, multiple myeloma,
merkel cell carcinoma, malignant mesothelioma, malignant fibrous
histiocytoma of bone and osteosarcoma, nasal cavity and paranasal
sinus cancer, nasopharyngeal cancer, neuroblastoma, non-Hodgkin's
lymphoma, non-small cell lung cancer (NSCLC), oral cancer, lip and
oral cavity cancer, oropharyngeal cancer, ovarian cancer,
pancreatic cancer, papillomatosis, paraganglioma, paranasal sinus
and nasal cavity cancer, parathyroid cancer, penile cancer,
pharyngeal cancer, pleuropulmonary blastoma, primary central
nervous system (CNS) lymphoma, prostate cancer, rectal cancer,
transitional cell cancer, retinoblastoma, rhabdomyosarcoma,
salivary gland cancer, skin cancer, stomach (gastric) cancer, small
cell lung cancer, small intestine cancer, soft tissue sarcoma,
T-Cell lymphoma, testicular cancer, throat cancer, thymoma and
thymic carcinoma, thyroid cancer, transitional cell cancer of the
renal pelvis and ureter, trophoblastic tumor, unusual cancers of
childhood, urethral cancer, uterine sarcoma, vaginal cancer, vulvar
cancer, or Viral-Induced cancer. In some embodiments, said method
relates to the treatment of a non-cancerous hyperproliferative
disorder such as benign hyperplasia of the skin (e. g., psoriasis),
restenosis, or prostate (e. g., benign prostatic hypertrophy
(BPH)).
[0150] In certain particular embodiments, the invention relates to
methods for treatment of lung cancers, the methods comprise
administering an effective amount of a KRAS, HRAS or NRAS G12C
mutant modulating compound and an additional therapeutic agent to a
subject in need thereof. In certain embodiments the lung cancer is
a non-small cell lung carcinoma (NSCLC), for example
adenocarcinoma, squamous-cell lung carcinoma or large-cell lung
carcinoma. In other embodiments, the lung cancer is a small cell
lung carcinoma. Other lung cancers treatable with the disclosed
compounds include, but are not limited to, glandular tumors,
carcinoid tumors and undifferentiated carcinomas.
[0151] Subjects that can be treated with the methods of this
invention include, for example, subjects that have been diagnosed
as having acute myeloid leukemia, acute myeloid leukemia, cancer in
adolescents, adrenocortical carcinoma childhood, AIDS-related
cancers (e.g. Lymphoma and Kaposi's Sarcoma), anal cancer, appendix
cancer, astrocytomas, atypical teratoid, basal cell carcinoma, bile
duct cancer, bladder cancer, bone cancer, brain stem glioma, brain
tumor, breast cancer, bronchial tumors, burkitt lymphoma, carcinoid
tumor, atypical teratoid, embryonal tumors, germ cell tumor,
primary lymphoma, cervical cancer, childhood cancers, chordoma,
cardiac tumors, chronic lymphocytic leukemia (CLL), chronic
myelogenous leukemia (CML), chronic myeloproliferative disorders,
colon cancer, colorectal cancer, craniopharyngioma, cutaneous
T-cell lymphoma, extrahepatic ductal carcinoma in situ (DCIS),
embryonal tumors, CNS cancer, endometrial cancer, ependymoma,
esophageal cancer, esthesioneuroblastoma, Ewing's sarcoma,
extracranial germ cell tumor, extragonadal germ cell tumor, eye
cancer, fibrous histiocytoma of bone, gall bladder cancer, gastric
cancer, gastrointestinal carcinoid tumor, gastrointestinal stromal
tumors (GIST), germ cell tumor, gestational trophoblastic tumor,
hairy cell leukemia, head and neck cancer, heart cancer, liver
cancer, Hodgkin's lymphoma, hypopharyngeal cancer, intraocular
melanoma, islet cell tumors, pancreatic neuroendocrine tumors,
kidney cancer, laryngeal cancer, lip and oral cavity cancer, liver
cancer, lobular carcinoma in situ (LCIS), lung cancer, lymphoma,
metastatic squamous neck cancer with occult primary, midline tract
carcinoma, mouth cancer multiple endocrine neoplasia syndromes,
multiple myeloma/plasma cell neoplasm, mycosis fungoides,
myelodysplastic syndromes, myelodysplastic/myeloproliferative
neoplasms, multiple myeloma, merkel cell carcinoma, malignant
mesothelioma, malignant fibrous histiocytoma of bone and
osteosarcoma, nasal cavity and paranasal sinus cancer,
nasopharyngeal cancer, neuroblastoma, non-Hodgkin's lymphoma,
non-small cell lung cancer (NSCLC), oral cancer, lip and oral
cavity cancer, oropharyngeal cancer, ovarian cancer, pancreatic
cancer, papillomatosis, paraganglioma, paranasal sinus and nasal
cavity cancer, parathyroid cancer, penile cancer, pharyngeal
cancer, pleuropulmonary blastoma, primary central nervous system
(CNS) lymphoma, prostate cancer, rectal cancer, transitional cell
cancer, retinoblastoma, rhabdomyosarcoma, salivary gland cancer,
skin cancer, stomach (gastric) cancer, small cell lung cancer,
small intestine cancer, soft tissue sarcoma, T-Cell lymphoma,
testicular cancer, throat cancer, thymoma and thymic carcinoma,
thyroid cancer, transitional cell cancer of the renal pelvis and
ureter, trophoblastic tumor, unusual cancers of childhood, urethral
cancer, uterine sarcoma, vaginal cancer, vulvar cancer, or
Viral-Induced cancer. In some embodiments subjects that are treated
according to the methods of the invention include subjects that
have been diagnosed as having a non-cancerous hyperproliferative
disorder such as benign hyperplasia of the skin (e. g., psoriasis),
restenosis, or prostate (e. g., benign prostatic hypertrophy
(BPH)).
[0152] The invention further provides methods of modulating a G12C
Mutant KRAS, HRAS or NRAS protein activity by contacting the
protein with an effective amount of a KRAS, HRAS or NRAS G12C
mutant modulating compound and an additional therapeutic agent.
Modulation can be inhibiting or activating protein activity. In
some embodiments, the invention provides methods of inhibiting
protein activity by contacting the G12C Mutant KRAS, HRAS or NRAS
protein with an effective amount of a KRAS, HRAS or NRAS G12C
mutant modulating compound and an additional therapeutic agent in
solution. In some embodiments, the invention provides methods of
inhibiting the G12C Mutant KRAS, HRAS or NRAS protein activity by
contacting a cell, tissue, organ that express the protein of
interest with a KRAS, HRAS or NRAS G12C mutant modulating compound
and an additional therapeutic agent. In some embodiments, the
invention provides methods of inhibiting protein activity in
subject including but not limited to rodents and mammal (e.g.,
human) by administering into the subject an effective amount of a
KRAS, HRAS or NRAS G12C mutant modulating compound and an
additional therapeutic agent. In some embodiments, the percentage
modulation exceeds 25%, 30%, 40%, 50%, 60%, 70%, 80%, or 90%. In
some embodiments, the percentage of inhibiting exceeds 25%, 30%,
40%, 50%, 60%, 70%, 80%, or 90%.
[0153] In some embodiments, the invention provides methods of
inhibiting KRAS, HRAS or NRAS G12C activity in a cell by contacting
said cell with an amount of a KRAS, HRAS or NRAS G12C mutant
modulating compound and an additional therapeutic agent sufficient
to inhibit the activity of KRAS, HRAS or NRAS G12C in said cell. In
some embodiments, the invention provides methods of inhibiting
KRAS, HRAS or NRAS G12C activity in a tissue by contacting said
tissue with an amount of a KRAS, HRAS or NRAS G12C mutant
modulating compound and an additional therapeutic agent sufficient
to inhibit the activity of KRAS, HRAS or NRAS G12C in said tissue.
In some embodiments, the invention provides methods of inhibiting
KRAS, HRAS or NRAS G12C activity in an organism by contacting said
organism with an amount of a KRAS, HRAS or NRAS G12C mutant
modulating compound and an additional therapeutic agent sufficient
to inhibit the activity of KRAS, HRAS or NRAS G12C in said
organism. In some embodiments, the invention provides methods of
inhibiting KRAS, HRAS or NRAS G12C activity in an animal by
contacting said animal with an amount of a KRAS, HRAS or NRAS G12C
mutant modulating compound and an additional therapeutic agent
sufficient to inhibit the activity of KRAS, HRAS or NRAS G12C in
said animal. In some embodiments, the invention provides methods of
inhibiting KRAS, HRAS or NRAS G12C activity in a subject by
contacting said subject with an amount of a KRAS, HRAS or NRAS G12C
mutant modulating compound and an additional therapeutic agent
sufficient to inhibit the activity of KRAS, HRAS or NRAS G12C in
said subject. In some embodiments, the invention provides methods
of inhibiting KRAS, HRAS or NRAS G12C activity in a human by
contacting said human with an amount of a KRAS, HRAS or NRAS G12C
mutant modulating compound and an additional therapeutic agent
sufficient to inhibit the activity of KRAS, HRAS or NRAS G12C in
said human. The present invention provides methods of treating a
disease mediated by KRAS, HRAS or NRAS G12C activity in a subject
in need of such treatment.
[0154] The additional therapeutic agent can be selected from any
number of therapeutic agents useful for treating cancer. Such
therapeutic agents can be approved for use in humans or
experimental. In some embodiments, the additional therapeutic agent
is selected from the group consisting of mitotic inhibitors,
alkylating agents, anti-metabolites, intercalating antibiotics,
growth factor inhibitors, cell cycle inhibitors, enzymes,
topoisomerase inhibitors, biological response modifiers,
anti-hormones, angiogenesis inhibitors, and anti-androgens.
[0155] In addition to the above examples, other non-limiting
examples of additional therapeutic agents useful in the described
methods are chemotherapeutic agents, cytotoxic agents, and
non-peptide small molecules such as Gleevec.RTM. (Imatinib
Mesylate), Velcade.RTM. (bortezomib), Casodex (bicalutamide),
Iressa.RTM. (gefitinib), and Adriamycin as well as a host of
chemotherapeutic agents. Non-limiting examples of chemotherapeutic
agents include alkylating agents such as thiotepa and
cyclosphosphamide (CYTOXAN.TM.); alkyl sulfonates such as busulfan,
improsulfan and piposulfan; aziridines such as benzodopa,
carboquone, meturedopa, and uredopa; ethylenimines and
methylamelamines including altretamine, triethylenemelamine,
triethylenephosphoramide, triethylenethiophosphaoramide and
trimethylolomelamine; nitrogen mustards such as chlorambucil,
chlornaphazine, cholophosphamide, estramustine, ifosfamide,
mechlorethamine, mechlorethamine oxide hydrochloride, melphalan,
novembichin, phenesterine, prednimustine, trofosfamide, uracil
mustard; nitrosureas such as carmustine, chlorozotocin,
fotemustine, lomustine, nimustine, ranimustine; antibiotics such as
aclacinomysins, actinomycin, authramycin, azaserine, bleomycins,
cactinomycin, calicheamicin, carabicin, carminomycin,
carzinophilin, Casodex.TM., chromomycins, dactinomycin,
daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin,
epirubicin, esorubicin, idarubicin, marcellomycin, mitomycins,
mycophenolic acid, nogalamycin, olivomycins, peplomycin,
potfiromycin, puromycin, quelamycin, rodorubicin, streptonigrin,
streptozocin, tubercidin, ubenimex, zinostatin, zorubicin;
anti-metabolites such as methotrexate and 5-fluorouracil (5-FU);
folic acid analogues such as denopterin, methotrexate, pteropterin,
trimetrexate; purine analogs such as fludarabine, 6-mercaptopurine,
thiamiprine, thioguanine; pyrimidine analogs such as ancitabine,
azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine,
doxifluridine, enocitabine, floxuridine, androgens such as
calusterone, dromostanolone propionate, epitiostanol, mepitiostane,
testolactone; anti-adrenals such as aminoglutethimide, mitotane,
trilostane; folic acid replenisher such as frolinic acid;
aceglatone; aldophosphamide glycoside; aminolevulinic acid;
amsacrine; bestrabucil; bisantrene; edatraxate; defofamine;
demecolcine; diaziquone; elfomithine; elliptinium acetate;
etoglucid; gallium nitrate; hydroxyurea; lentinan; lonidamine;
mitoguazone; mitoxantrone; mopidamol; nitracrine; pentostatin;
phenamet; pirarubicin; podophyllinic acid; 2-ethylhydrazide;
procarbazine; PSK.RTM.; razoxane; sizofiran; spirogermanium;
tenuazonic acid; triaziquone; 2,2',2''-trichlorotriethylamine;
urethan; vindesine; dacarbazine; mannomustine; mitobronitol;
mitolactol; pipobroman; gacytosine; arabinoside ("Ara-C");
cyclophosphamide; thiotepa; taxanes, e.g. paclitaxel (TAXOL.TM.,
Bristol-Myers Squibb Oncology, Princeton, N.J.) and docetaxel
(TAXOTERE.TM., Rhone-Poulenc Rorer, Antony, France); retinoic acid;
esperamicins; capecitabine; and pharmaceutically acceptable salts,
acids or derivatives of any of the above.
[0156] Also included as suitable chemotherapeutic cell conditioners
are anti-hormonal agents that act to regulate or inhibit hormone
action on tumors such as anti-estrogens including for example
tamoxifen, (Nolvadex.TM.), raloxifene, aromatase inhibiting
4(5)-imidazoles, 4-hydroxytamoxifen, trioxifene, keoxifene, LY
117018, onapristone, and toremifene (Fareston); and anti-androgens
such as flutamide, nilutamide, bicalutamide, leuprolide, and
goserelin; chlorambucil; gemcitabine; 6-thioguanine;
mercaptopurine; methotrexate; platinum analogs such as cisplatin
and carboplatin; vinblastine; platinum; etoposide (VP-16);
ifosfamide; mitomycin C; mitoxantrone; vincristine; vinorelbine;
navelbine; novantrone; teniposide; daunomycin; aminopterin; xeloda;
ibandronate; camptothecin-11 (CPT-11); topoisomerase inhibitor RFS
2000; difluoromethylornithine (DMFO). Where desired, the compounds
or pharmaceutical composition of the present invention can be used
in combination with commonly prescribed anti-cancer drugs such as
Herceptin.RTM., Avastin.RTM., Erbitux.RTM., Rituxan.RTM.,
Taxol.RTM., Arimidex.RTM., Taxotere.RTM., ABVD, AVICINE,
Abagovomab, Acridine carboxamide, Adecatumumab,
17-N-Allylamino-17-demethoxygeldanamycin, Alpharadin, Alvocidib,
3-Aminopyridine-2-carboxaldehyde thiosemicarbazone, Amonafide,
Anthracenedione, Anti-CD22 immunotoxins, Antineoplastic,
Antitumorigenic herbs, Apaziquone, Atiprimod, Azathioprine,
Belotecan, Bendamustine, BIBW 2992, Biricodar, Brostallicin,
Bryostatin, Buthionine sulfoximine, CBV (chemotherapy), Calyculin,
cell-cycle nonspecific antineoplastic agents, Dichloroacetic acid,
Discodermolide, Elsamitrucin, Enocitabine, Epothilone, Eribulin,
Everolimus, Exatecan, Exisulind, Ferruginol, Forodesine,
Fosfestrol, ICE chemotherapy regimen, IT-101, Imexon, Imiquimod,
Indolocarbazole, Irofulven, Laniquidar, Larotaxel, Lenalidomide,
Lucanthone, Lurtotecan, Mafosfamide, Mitozolomide, Nafoxidine,
Nedaplatin, Olaparib, Ortataxel, PAC-1, Pawpaw, Pixantrone,
Proteasome inhibitor, Rebeccamycin, Resiquimod, Rubitecan, SN-38,
Salinosporamide A, Sapacitabine, Stanford V, Swainsonine,
Talaporfin, Tariquidar, Tegafur-uracil, Temodar, Tesetaxel,
Triplatin tetranitrate, Tris(2-chloroethyl)amine, Troxacitabine,
Uramustine, Vadimezan, Vinflunine, ZD6126 or Zosuquidar.
[0157] This invention further relates to a method for using a KRAS,
HRAS or NRAS G12C mutant modulating compound and an additional
therapeutic agent, in combination with radiation therapy for
inhibiting abnormal cell growth or treating the hyperproliferative
disorder in the subject. Techniques for administering radiation
therapy are known in the art, and these techniques can be used in
the combination therapy described herein. The administration of the
a KRAS, HRAS or NRAS G12C mutant modulating compound and an
additional therapeutic agent in this combination therapy can be
determined as described herein.
[0158] Radiation therapy can be administered through one of several
methods, or a combination of methods, including without limitation
external-beam therapy, internal radiation therapy, implant
radiation, stereotactic radiosurgery, systemic radiation therapy,
radiotherapy and permanent or temporary interstitial brachytherapy.
The term "brachytherapy," as used herein, refers to radiation
therapy delivered by a spatially confined radioactive material
inserted into the body at or near a tumor or other proliferative
tissue disease site. The term is intended without limitation to
include exposure to radioactive isotopes (e.g. At-211, I-131,
I-125, Y-90, Re-186, Re-188, Sm-153, Bi-212, P-32, and radioactive
isotopes of Lu). Suitable radiation sources for use as a cell
conditioner of the present invention include both solids and
liquids. By way of non-limiting example, the radiation source can
be a radionuclide, such as I-125, I-131, Yb-169, Ir-192 as a solid
source, I-125 as a solid source, or other radionuclides that emit
photons, beta particles, gamma radiation, or other therapeutic
rays. The radioactive material can also be a fluid made from any
solution of radionuclide(s), e.g., a solution of I-125 or I-131, or
a radioactive fluid can be produced using a slurry of a suitable
fluid containing small particles of solid radionuclides, such as
Au-198, Y-90. Moreover, the radionuclide(s) can be embodied in a
gel or radioactive micro spheres.
[0159] Without being limited by any theory, the methods of the
present invention can render abnormal cells more sensitive to
treatment with radiation for purposes of killing and/or inhibiting
the growth of such cells. Accordingly, this invention further
relates to a method for sensitizing abnormal cells in a subject to
treatment with radiation which comprises administering to the
subject an amount of a KRAS, HRAS or NRAS G12C mutant modulating
compound and an additional therapeutic agent, which amount is
effective is sensitizing abnormal cells to treatment with
radiation. The amount of the KRAS, HRAS or NRAS G12C mutant
modulating compound and an additional therapeutic agent in this
method can be determined according to the means for ascertaining
effective amounts known in the art.
[0160] In other embodiments, the additional therapeutic agent is
selected from anti-angiogenesis agents, signal transduction
inhibitors, antiproliferative agents, glycolysis inhibitors, or
autophagy inhibitors.
[0161] Anti-angiogenesis agents, such as MMP-2
(matrix-metalloproteinase 2) inhibitors, MMP-9
(matrix-metalloproteinase 9) inhibitors, and COX-11 (cyclooxygenase
11) inhibitors, can be used as the additional therapeutic agent in
the methods described herein. Anti-angiogenesis agents include, for
example, rapamycin, temsirolimus (CCI-779), everolimus (RAD001),
sorafenib, sunitinib, and bevacizumab. Examples of useful COX-II
inhibitors include CELEBREX.TM. (alecoxib), valdecoxib, and
rofecoxib.
[0162] Examples of useful matrix metalloproteinase inhibitors are
described in WO 96/33172 (published Oct. 24,1996), WO 96/27583
(published Mar. 7,1996), European Patent Application No. 97304971.1
(filed Jul. 8,1997), European Patent Application No. 99308617.2
(filed Oct. 29, 1999), WO 98/07697 (published Feb. 26,1998), WO
98/03516 (published Jan. 29,1998), WO 98/34918 (published Aug. 13,
1998), WO 98/34915 (published Aug. 13, 1998), WO 98/33768
(published Aug. 6, 1998), WO 98/30566 (published Jul. 16, 1998),
European Patent Publication 606,046 (published Jul. 13,1994),
European Patent Publication 931, 788 (published Jul. 28,1999), WO
90/05719 (published May 31, 1990), WO 99/52910 (published Oct.
21,1999), WO 99/52889 (published Oct. 21, 1999), WO 99/29667
(published Jun. 17,1999), PCT International Application No.
PCT/IB98/01113 (filed Jul. 21,1998), European Patent Application
No. 99302232.1 (filed Mar. 25,1999), Great Britain Patent
Application No. 9912961.1 (filed Jun. 3, 1999), U.S. Provisional
Application No. 60/148,464 (filed Aug. 12,1999), U.S. Pat. No.
5,863,949 (issued Jan. 26,1999), U.S. Pat. No. 5,861,510 (issued
Jan. 19,1999), and European Patent Publication 780,386 (published
Jun. 25, 1997), all of which are incorporated herein in their
entireties by reference. Preferred MMP-2 and MMP-9 inhibitors are
those that have little or no activity inhibiting MMP-1. More
preferred, are those that selectively inhibit MMP-2 and/or AMP-9
relative to the other matrix-metalloproteinases (i. e., MAP-1,
MMP-3, MMP-4, MMP-5, MMP-6, MMP-7, MMP-8, MMP-10, MMP-11, MMP-12,
and MMP-13). Some specific examples of MMP inhibitors useful in the
invention are AG-3340, RO 32-3555, and RS 13-0830.
[0163] Autophagy inhibitors include, but are not limited to
chloroquine, 3-methyladenine, hydroxychloroquine (Plaquenil.TM.),
bafilomycin A1, 5-amino-4-imidazole carboxamide riboside (AICAR),
okadaic acid, autophagy-suppressive algal toxins which inhibit
protein phosphatases of type 2A or type 1, analogues of cAMP, and
drugs which elevate cAMP levels such as adenosine, LY204002,
N6-mercaptopurine riboside, and vinblastine. In addition, antisense
or siRNA that inhibits expression of proteins including but not
limited to ATG5 (which are implicated in autophagy), may also be
used.
B. Compounds
[0164] As noted above, embodiments of the present methods include
administration of a KRAS, HRAS or NRAS G12C mutant modulating
compound ("compound"). The compounds have activity as modulators of
KRAS, HRAS or NRAS G12C mutant protein activity. In some
embodiments, the compound is a KRAS, HRAS or NRAS G12C mutant
modulating compound. In an aspect, the compounds are capable of
selectively binding to and/or modulating a G12C mutant KRAS, HRAS
or NRAS protein. The compounds may modulate the G12C mutant KRAS,
HRAS or NRAS protein by reaction with an amino acid. While not
wishing to be bound by theory, the present applicants believe that,
in some embodiments, the compounds selectively react with the G12C
mutant KRAS, HRAS or NRAS proteins by forming a covalent bond with
the cysteine at the 12 position of a G12C mutant KRAS, HRAS or NRAS
protein. By binding to the Cystine 12, the compounds may lock the
switch II of the G12C mutant KRAS, HRAS or NRAS into an inactive
stage. This inactive stage may be distinct from those observed for
GTP and GDP bound KRAS, HRAS or NRAS. Some of the compounds may
also be able to perturb the switch I conformation. Some of the
compounds may favor the binding of the bound KRAS, HRAS or NRAS to
GDP rather than GTP and therefore sequester the KRAS, HRAS or NRAS
into an inactive KRAS, HRAS or NRAS GDP state. Because effector
binding to KRAS, HRAS or NRAS is highly sensitive to the
conformation of switch I and II, the irreversible binding of these
compounds may disrupt KRAS, HRAS or NRAS downstream signaling.
[0165] The present methods are not limited by the exact structure
of the KRAS, HRAS or NRAS G12C mutant modulating compound, provided
it has the above noted functionality (e.g., modulating KRAS, HRAS
or NRAS G12C mutant protein activity). Examples of compounds useful
in certain embodiments of the methods are provided in PCT Pub. Nos.
WO 2013/155223 and 2015/054572, the compounds and methods of which
are incorporated herein by reference in their entirety. Other
compounds useful in different embodiments of the method are
provided herein below.
1. Compounds of Structure (I)
[0166] As noted above, in one embodiment of the present invention,
compounds having activity as modulators of a G12C mutant KRAS, HRAS
or NRAS protein are provided, the compounds have the following
structure (I):
##STR00001##
or a pharmaceutically acceptable salt, tautomer, prodrug or
stereoisomer thereof, wherein:
[0167] A is CR.sup.1, CR.sup.2b, NR.sup.5 or S;
[0168] B is a bond, CR.sup.1 or CR.sup.2c
[0169] G.sup.1 and G.sup.2 are each independently N or CH;
[0170] W, X and Y are each independently N, NR.sup.5 or
CR.sup.6;
[0171] Z is a bond, N or CR.sup.6;
[0172] L.sup.1 is a bond or NR.sup.7;
[0173] L.sup.2 is a bond or alkylene;
[0174] R.sup.1 is H, cyano, halo, C.sub.1-C.sub.6alkyl,
C.sub.1-C.sub.6alkylaminyl, C.sub.3-C.sub.8 cycloalkyl,
C.sub.1-C.sub.6alkenyl or C.sub.3-C.sub.8cycloalkenyl,
heterocyclyl, heteroaryl, aryloxy or aryl;
[0175] R.sup.2a, R.sup.2b and R.sup.2C are each independently H,
halo, hydroxyl, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 haloalkyl,
C.sub.1-C.sub.6 alkoxy, C.sub.3-C.sub.8 cycloalkyl or aryl;
[0176] R.sup.3a and R.sup.3b are, at each occurrence, independently
H, --OH, --NH.sub.2, --CO.sub.2H, halo, cyano, C.sub.1-C.sub.6
alkyl, C.sub.1-C.sub.6 alkynyl, hydroxylalkly, aminylalkyl,
alkylaminylalkyl, cyanoalkyl, carboxyalkyl, aminylcarbonylalkyl or
aminylcarbonyl; or R.sup.3a and R.sup.3b join to form a carbocyclic
or heterocyclic ring; or R.sup.3a is H, --OH, --NH.sub.2,
--CO.sub.2H, halo, cyano, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6
alkynyl, hydroxylalkly, aminylalkyl, alkylaminylalkyl, cyanoalkyl,
carboxyalkyl, aminylcarbonylalkyl or aminylcarbonyl, and R.sup.3b
joins with R.sup.4b to form a carbocyclic or heterocyclic ring;
[0177] R.sup.4a and R.sup.4b are, at each occurrence, independently
H, --OH, --NH.sub.2, --CO.sub.2H, halo, cyano, C.sub.1-C.sub.6
alkyl, C.sub.1-C.sub.6 alkynyl, hydroxylalkly, aminylalkyl,
alkylaminylalkyl, cyanoalkyl, carboxyalkyl, aminylcarbonylalkyl or
aminylcarbonyl; or R.sup.4a and R.sup.4b join to form a carbocyclic
or heterocyclic ring; or R.sup.4a is H, --OH, --NH.sub.2,
--CO.sub.2H, halo, cyano, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6
alkynyl, hydroxylalkly, aminylalkyl, alkylaminylalkyl, cyanoalkyl,
carboxyalkyl, aminylcarbonylalkyl or aminylcarbonyl, and R.sup.4b
joins with R.sup.3b to form a carbocyclic or heterocyclic ring;
[0178] R.sup.5 is, at each occurrence, independently H,
C.sub.1-C.sub.6 alkyl or a bond to L.sup.1;
[0179] R.sup.6 is, at each occurrence, independently H, oxo, cyano,
cyanoalkyl, amino, aminylalkyl, aminylalkylaminyl, aminylcarbonyl,
alkylaminyl, haloalkylaminyl, hydroxylalkyaminyl, amindinylalkyl,
amidinylalkoxy, amindinylalkylaminyl, guanidinylalkyl,
guanidinylalkoxy, guanidinylalkylaminyl, C.sub.1-C.sub.6 alkoxy,
aminylalkoxy, alkylcarbonylaminylalkoxy, C.sub.1-C.sub.6 alkyl,
heterocyclyl, heterocyclyloxy, heterocyclylalkyloxy,
heterocyclylaminyl, heterocyclylalkylaminyl, heteroaryl,
heteroaryloxy, heteroarylalkyloxy, heteroarylaminyl,
heteroarylalkylaminyl, aryl, aryloxy, arylamino, arylalkylamino,
arylalkyloxy or a bond to L.sup.1;
[0180] R.sup.7 is H or C.sub.1-C.sub.6 alkyl;
[0181] m.sup.1 and m.sup.2 are each independently 1, 2 or 3;
[0182] indicates a single or double bond such that all valences are
satisfied; and
[0183] E is an electrophilic moiety capable of forming a covalent
bond with the cysteine residue at position 12 of a KRAS, HRAS or
NRAS G12C mutant protein,
[0184] wherein at least one of W, X, Y or Z is CR.sup.6 where
R.sup.6 is a bond to L.sup.1.
[0185] In some embodiments when R.sup.1, R.sup.2a, R.sup.2b and
R.sup.2c are all independently selected from H and halo, then X and
Z are both N and at least one of R.sup.3a, R.sup.3b, R.sup.4a or
R.sup.4b is not H, and provided that at least one of R.sup.2a,
R.sup.2b or R.sup.2c is not H when R.sup.1 is pyridyl.
[0186] In other embodiments, the compound has the following
structure (I):
##STR00002##
[0187] or a pharmaceutically acceptable salt, tautomer, prodrug or
stereoisomer thereof, wherein:
[0188] A is CR.sup.1, CR.sup.2b, NR.sup.7 or S;
[0189] B is a bond, CR.sup.1 or CR.sup.2c
[0190] G.sup.1 and G.sup.2 are each independently N or CH;
[0191] W, X and Y are each independently N, NR.sup.5 or
CR.sup.6;
[0192] Z is a bond, N or CR.sup.6a or Z is NH when Y is
C.dbd.O;
[0193] L.sup.1 is a bond or NR.sup.7;
[0194] L.sup.2 is a bond or alkylene;
[0195] R.sup.1 is H, cyano, halo, heterocyclyl, heteroaryl, aryloxy
or aryl;
[0196] R.sup.2a, R.sup.2b and R.sup.2C are each independently H,
halo, hydroxyl, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 haloalkyl,
C.sub.1-C.sub.6 alkoxy, C.sub.3-C.sub.8 cycloalkyl or aryl;
[0197] R.sup.3a and R.sup.3b are, at each occurrence, independently
H, --OH, --NH.sub.2, --CO.sub.2H, halo, cyano, C.sub.1-C.sub.6
alkyl, C.sub.1-C.sub.6 alkynyl, hydroxylalkly, aminylalkyl,
alkylaminylalkyl, cyanoalkyl, carboxyalkyl, aminocarbonylalkyl or
aminocarbonyl; or R.sup.3a and R.sup.3b join to form a carbocyclic
or heterocyclic ring; or R.sup.3a is H, --OH, --NH.sub.2,
--CO.sub.2H, halo, cyano, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6
alkynyl, hydroxylalkly, aminoalkyl, alkylaminylalkyl, cyanoalkyl,
carboxyalkyl, aminylcarbonylalkyl or aminylcarbonyl, and R.sup.3b
joins with R.sup.4b to form a carbocyclic or heterocyclic ring;
[0198] R.sup.4a and R.sup.4b are, at each occurrence, independently
H, --OH, --NH.sub.2, --CO.sub.2H, halo, cyano, C.sub.1-C.sub.6
alkyl, C.sub.1-C.sub.6 alkynyl, hydroxylalkly, aminylalkyl,
alkylaminylalkyl, cyanoalkyl, carboxyalkyl, aminocarbonylalkyl or
aminocarbonyl; or R.sup.4a and R.sup.4b join to form a carbocyclic
or heterocyclic ring; or R.sup.4a is H, --OH, --NH.sub.2,
--CO.sub.2H, halo, cyano, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6
alkynyl, hydroxylalkly, aminylalkyl, alkylaminoalkyl, cyanoalkyl,
carboxyalkyl, aminylcarbonylalkyl or aminylcarbonyl, and R.sup.4b
joins with R.sup.3b to form a carbocyclic or heterocyclic ring;
[0199] R.sup.5 is, at each occurrence, independently H,
C.sub.1-C.sub.6 alkyl or a bond to L.sup.1;
[0200] R.sup.6 is, at each occurrence, independently H, oxo, cyano,
cyanoalkyl, amino, aminylalkyl, aminylalkylaminyl, aminocarbonyl,
alkylaminyl, haloalkylamino, hydroxylalkyamino, amindinylalkyl,
amidinylalkoxy, amindinylalkylaminyl, guanidinylalkyl,
guanidinylalkoxy, guanidinylalkylaminyl, C.sub.1-C.sub.6 alkoxy,
aminylalkoxy, alkylcarbonylaminylalkoxy, C.sub.1-C.sub.6 alkyl,
heterocyclyl, heterocyclyloxy, heterocyclylalkyloxy,
heterocyclylamino, heterocyclylalkylamino, heteroaryl,
heteroaryloxy, heteroarylalkyloxy, heteroarylamino,
heteroarylalkylamino, aryl, aryloxy, arylamino, arylalkylamino,
arylalkyloxy or a bond to L.sup.1;
[0201] R.sup.6a is H, alkyl or a bond to L.sup.1;
[0202] R.sup.7 is H or C.sub.1-C.sub.6 alkyl
[0203] m.sup.1 and m.sup.2 are each independently 1, 2 or 3;
[0204] indicates a single or double bond such that all valences are
satisfied; and
[0205] E is an electrophilic moiety capable of forming a covalent
bond with the cysteine residue at position 12 of a KRAS, HRAS or
NRAS G12C mutant protein,
[0206] wherein at least one of W, X, Y or Z is CR.sup.6 where
R.sup.6 is a bond to L.sup.1 or at least one of W, X or Y is
NR.sup.5, wherein R.sup.5 is a bond to L.sup.1.
[0207] In some embodiments of the compound of structure (I), the
bond between W and X is a double bond. In other embodiments, the
bond between Y and Z is a double bond. In more embodiments, the
bond between A and B is a double bond. In still more embodiments,
the bonds between W and X, Y and Z and A and B are each double
bonds.
[0208] In some more embodiments of the foregoing compound of
structure (I):
[0209] A is CR.sup.1, CR.sup.2b, NR.sup.7 or S;
[0210] B is a bond, CR.sup.1 or CR.sup.2c
[0211] G.sup.1 and G.sup.2 are each independently N or CH;
[0212] W, X and Y are each independently N, NR.sup.5 or
CR.sup.6;
[0213] Z is a bond, N or CR.sup.6;
[0214] L.sup.1 is a bond or NR.sup.7;
[0215] L.sup.2 is a bond or alkylene;
[0216] R.sup.1 is H, cyano, halo, heterocyclyl, heteroaryl, aryloxy
or aryl;
[0217] R.sup.2a, R.sup.2b and R.sup.2C are each independently H,
halo, hydroxyl, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 haloalkyl
C.sub.3-C.sub.8 cycloalkyl or aryl;
[0218] R.sup.3a and R.sup.3b are, at each occurrence, independently
H, --OH, --NH.sub.2, --CO.sub.2H, halo, cyano, C.sub.1-C.sub.6
alkyl, C.sub.1-C.sub.6 alkynyl, hydroxylalkly, aminylalkyl,
alkylaminylalkyl, cyanoalkyl, carboxyalkyl, aminylcarbonylalkyl or
aminylcarbonyl; or R.sup.3a and R.sup.3b join to form a carbocyclic
or heterocyclic ring; or R.sup.3a is H, --OH, --NH.sub.2,
--CO.sub.2H, halo, cyano, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6
alkynyl, hydroxylalkly, aminylalkyl, alkylaminylalkyl, cyanoalkyl,
carboxyalkyl, aminylcarbonylalkyl or aminylcarbonyl, and R.sup.3b
joins with R.sup.4b to form a carbocyclic or heterocyclic ring;
[0219] R.sup.4a and R.sup.4b are, at each occurrence, independently
H, --OH, --NH.sub.2, --CO.sub.2H, halo, cyano, C.sub.1-C.sub.6
alkyl, C.sub.1-C.sub.6 alkynyl, hydroxylalkly, aminylalkyl,
alkylaminylalkyl, cyanoalkyl, carboxyalkyl, aminylcarbonylalkyl or
aminylcarbonyl; or R.sup.4a and R.sup.4b join to form a carbocyclic
or heterocyclic ring; or R.sup.4a is H, --OH, --NH.sub.2,
--CO.sub.2H, halo, cyano, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6
alkynyl, hydroxylalkly, aminylalkyl, alkylaminylalkyl, cyanoalkyl,
carboxyalkyl, aminylcarbonylalkyl or aminylcarbonyl, and R.sup.4b
joins with R.sup.3b to form a carbocyclic or heterocyclic ring;
[0220] R.sup.5 and R.sup.7 are each independently H or
C.sub.1-C.sub.6 alkyl;
[0221] R.sup.6 is, at each occurrence, independently H, oxo, cyano,
cyanoalkyl, amino, aminylcarbonyl, alkylaminyl, C.sub.1-C.sub.6
alkoxy, C.sub.1-C.sub.6 alkyl or a bond to L.sup.1;
[0222] m.sup.1 and m.sup.2 are each independently 1, 2 or 3;
[0223] indicates a single or double bond such that all valences are
satisfied; and
[0224] E is an electrophilic moiety capable of forming a covalent
bond with the cysteine residue at position 12 of a KRAS, HRAS or
NRAS G12C mutant protein,
[0225] wherein at least one of W, X, Y or Z is CR.sup.6 where
R.sup.6 is a bond to L.sup.1, and
[0226] provided that when R.sup.1, R.sup.2a, R.sup.2b and R.sup.2c
are all independently selected from H and halo, then X and Z are
both N and at least one of R.sup.3a, R.sup.3b, R.sup.4a or R.sup.4b
is not H, and provided that at least one of R.sup.2a, R.sup.2b or
R.sup.2c is not H when R.sup.1 is pyridyl.
[0227] In some other embodiments of the foregoing compound of
structure (I):
[0228] A is CR.sup.2b, NR.sup.7 or S;
[0229] B is a bond or CR.sup.2c
[0230] G.sup.1 and G.sup.2 are each independently N or CH;
[0231] W, X and Y are each independently N, NR.sup.5 or
CR.sup.6;
[0232] Z is a bond, N or CR.sup.6;
[0233] L.sup.1 is a bond or NR.sup.7;
[0234] L.sup.2 is a bond or alkylene;
[0235] R.sup.1 is cyano, C.sub.1-C.sub.6alkyl,
C.sub.1-C.sub.6alkylaminyl, C.sub.3-C.sub.8 cycloalkyl,
C.sub.1-C.sub.6alkenyl or C.sub.3-C.sub.8 cycloalkenyl,
heterocyclyl or aryl;
[0236] R.sup.2a, R.sup.2b and R.sup.2C are each independently H,
halo, C.sub.1-C.sub.6 alkyl or C.sub.3-C.sub.8 cycloalkyl;
[0237] R.sup.3a and R.sup.3b are, at each occurrence, independently
H, --OH, --NH.sub.2, --CO.sub.2H, halo, cyano, hydroxylalkly,
aminylalkyl, cyanoalkyl, carboxyalkyl or aminylcarbonyl; or
R.sup.3a and R.sup.3b join to form a carbocyclic or heterocyclic
ring; or R.sup.3a is H, --OH, --NH.sub.2, --CO.sub.2H, halo, cyano,
hydroxylalkly, aminylalkyl, cyanoalkyl, carboxyalkyl or
aminylcarbonyl, and R.sup.3b joins with R.sup.4b to form a
carbocyclic or heterocyclic ring;
[0238] R.sup.4a and R.sup.4b are, at each occurrence, independently
H, --OH, --NH.sub.2, --CO.sub.2H, halo, cyano, hydroxylalkly,
aminylalkyl, cyanoalkyl, carboxyalkyl or aminylcarbonyl; or
R.sup.4a and R.sup.4b join to form a carbocyclic or heterocyclic
ring; or R.sup.4a is H, --OH, --NH.sub.2, --CO.sub.2H, halo, cyano,
hydroxylalkly, aminylalkyl, cyanoalkyl, carboxyalkyl or
aminylcarbonyl, and R.sup.4b joins with R.sup.3b to form a
carbocyclic or heterocyclic ring;
[0239] R.sup.5 and R.sup.7 are each independently H or
C.sub.1-C.sub.6alkyl;
[0240] R.sup.6 is, at each occurrence, independently H, cyano,
amino, alkylaminyl, C.sub.1-C.sub.6alkoxy, C.sub.1-C.sub.6alkyl or
a bond to L.sup.1;
[0241] m.sup.1 and m.sup.2 are each independently 1, 2 or 3;
[0242] indicates a single or double bond such that all valences are
satisfied; and
[0243] E is an electrophilic moiety capable of forming a covalent
bond with the cysteine residue at position 12 of a KRAS, HRAS or
NRAS G12C mutant protein,
[0244] wherein at least one of W, X or Y is CR.sup.6 where R.sup.6
is a bond to L.sup.1.
[0245] In still other embodiments of the foregoing compound of
structure (I), R.sup.1 is H, cyano, halo, heterocyclyl, heteroaryl,
aryloxy or aryl.
[0246] The structure of E is not particularly limited provided it
is capable of forming a covalent bond with a nucleophile, such as
the cysteine residue at position 12 of a KRAS, HRAS or NRAS G12C
mutant protein. Accordingly, moieties which are capable of reaction
with (e.g., by covalent bond formation) a nucleophile are
preferred. In certain embodiments, E is capable of reacting in a
conjugate addition manner (e.g., 1.4-conjugate addition) with an
appropriately reactive nucleophile. In some embodiments, E
comprises conjugated pi bonds such that delocalization of electrons
results in at least one atom (e.g., a carbon atom) having a
positive charge, partial positive charge or a polarized bond. In
other embodiments, E comprises one or more bonds wherein the
electronegativity of the two atoms forming the bonds is
sufficiently different such that a partial positive charge (e.g.,
by polarization of the bond) resides on one of the atoms, for
example on a carbon atom. E moieties comprising carbon-halogen
bonds, carbon-oxygen bonds or carbon bonds to various leaving
groups known in the art are examples of such E moieties.
[0247] In certain embodiments of the foregoing, E has the following
structure:
##STR00003##
wherein:
[0248] represents a double or triple bond;
[0249] Q is --C(.dbd.O)--, --C(.dbd.NR.sup.8')--,
--NR.sup.8C(.dbd.O)--, --S(.dbd.O).sub.2-- or
--NR.sup.8S(.dbd.O).sub.2--;
[0250] R.sup.8 is H, C.sub.1-C.sub.6alkyl or hydroxylalkyl;
[0251] R.sup.8 is H, --OH, --CN or C.sub.1-C.sub.6alkyl; and
[0252] when is a double bond then R.sup.9 and R.sup.10 are each
independently H, cyano, carboxyl, C.sub.1-C.sub.6alkyl,
alkoxycarbonyl, aminylalkyl, alkylaminylalkyl, or hydroxylalkyl or
R.sup.9 and R.sup.10 join to form a carbocyclic or heterocyclic
ring;
[0253] when is a triple bond; then R.sup.9 is absent and R.sup.10
is H, C.sub.1-C.sub.6alkyl, aminylalkyl, alkylaminylakyyl or
hydroxylalkyl.
[0254] In certain embodiments when is a double bond then R.sup.9
and R.sup.10 are each independently H, cyano, C.sub.1-C.sub.6alkyl,
aminylalkyl, alkylaminylalkyl, or hydroxylalkyl or R.sup.9 and
R.sup.10 join to form a carbocyclic or heterocyclic ring.
[0255] In some of the foregoing embodiments, Q is-C(.dbd.O)--,
--NR.sup.8C(.dbd.O)--, --S(.dbd.O).sub.2-- or
--NR.sup.8S(.dbd.O).sub.2--.
[0256] In some other of the foregoing embodiments, Q is
--C(.dbd.NR.sup.8')--, wherein R.sup.8' is H, --OH, --CN or
C.sub.1-C.sub.6alkyl. For example, in some embodiments R.sup.8 is
H. In other embodiments, R.sup.8' is --CN. In other embodiments,
R.sup.8' is --OH.
[0257] In some embodiments, the compound has the following
structure (I'):
##STR00004##
wherein R' is R.sup.1 and R'' is R.sup.2c or R' is H and R'' is
R.sup.1.
[0258] In other embodiments, the compound has the following
structure (I'a):
##STR00005##
wherein:
[0259] represents a double or triple bond;
[0260] Q is --C(.dbd.O)--, --C(.dbd.NR.sup.8')--,
--NR.sup.8C(.dbd.O)--, --S(.dbd.O).sub.2-- or
--NR.sup.8S(.dbd.O).sub.2--;
[0261] R.sup.8 is H, C.sub.1-C.sub.6alkyl or hydroxylalkyl;
[0262] R.sup.8' is H, --OH, --CN or C.sub.1-C.sub.6alkyl;
[0263] when is a double bond then R.sup.9 and R.sup.10 are each
independently H, cyano, carboxyl, C.sub.1-C.sub.6alkyl,
alkoxycarbonyl, aminylalkyl, alkylaminylalkyl, heteroaryl or
hydroxylalkyl or R.sup.9 and R.sup.10 join to form a carbocyclic or
heterocyclic ring;
[0264] when is a triple bond then R.sup.9 is absent and R.sup.10 is
H, C.sub.1-C.sub.6alkyl, aminylalkyl, alkylaminylalkyl or
hydroxylalkyl; and
[0265] R' is R.sup.1 and R'' is R.sup.2c or R' is H and R'' is
R.sup.1.
[0266] In some of the foregoing embodiments of compound (I'a), Q is
Q is --C(.dbd.O)--, --NR.sup.8C(.dbd.O)--, --S(.dbd.O).sub.2-- or
--NR.sup.8S(.dbd.O).sub.2--.
[0267] In some other of the foregoing embodiments of compound
(I'a), Q is --C(.dbd.NR)--, wherein R.sup.8 is H, --OH, --CN or
C.sub.1-C.sub.6alkyl. For example, in some embodiments R.sup.8' is
H. In other embodiments, R.sup.8' is --CN. In other embodiments,
R.sup.8' is --OH.
[0268] In still more embodiments of the foregoing compounds, the
compound has one of the following structures (I'b), (I'c), (I'd) or
(I'e):
##STR00006##
[0269] In still more embodiments, the compound has one of the
following structures (I'f), (I'g), (I'h) or (I'i):
##STR00007##
[0270] In some embodiments of the compounds of structures (I'f),
(I'g), (I'h) or (I'i), R.sup.1 is aryl and R.sup.2c and R.sup.2b
are independently selected from H and halo, for example in some
further embodiments R.sup.1 is aryl and R.sup.2c and R.sup.2b are
independently selected from halo.
[0271] In different embodiments, the compound has one of the
following structures (I'j), (I'k), (I'l) or (I'm):
##STR00008##
[0272] In some embodiments of the compounds of structures (I'j),
(I'k), (I'l) or (I'm), R.sup.1 is aryl and R.sup.2a and R.sup.2b
are independently selected from H and halo, for example in some
further embodiments R.sup.1 is aryl and R.sup.2a and R.sup.2b are
independently selected from halo.
[0273] In other embodiments, the compound has the following
structure (I''):
##STR00009##
wherein R' is R.sup.1 and R'' is R.sup.2c or R' is H and R'' is
R.sup.1. For example, in some embodiments the compound has the
following structure (I''a):
##STR00010##
wherein:
[0274] represents a double or triple bond;
[0275] Q is --C(.dbd.O)--, --C(.dbd.NR.sup.8')--,
--NR.sup.8C(.dbd.O)--, --S(.dbd.O).sub.2-- or
--NR.sup.8S(.dbd.O).sub.2--;
[0276] R.sup.8 is H, C.sub.1-C.sub.6alkyl or hydroxylalkyl;
[0277] R.sup.8' is H, --OH, --CN or C.sub.1-C.sub.6alkyl;
[0278] when is a double bond then R.sup.9 and R.sup.10 are each
independently H, cyano, carboxyl, C.sub.1-C.sub.6alkyl,
alkoxycarbonyl, aminylalkyl, alkylaminylalkyl, heteroaryl or
hydroxylalkyl or R.sup.9 and R.sup.10 join to form a carbocyclic or
heterocyclic ring;
[0279] when is a triple bond then R.sup.9 is absent and R.sup.10 is
H, C.sub.1-C.sub.6alkyl, aminylalkyl, alkylaminylalkyl or
hydroxylalkyl; and
[0280] R' is R.sup.1 and R'' is R.sup.2c or R' is H and R'' is
R.sup.1.
[0281] In some of the foregoing embodiments of compound (I''a), Q
is Q is --C(.dbd.O)--, --NR.sup.8C(.dbd.O)--, --S(.dbd.O).sub.2--
or --NR.sup.8S(.dbd.O).sub.2--.
[0282] In some other of the foregoing embodiments of compound
(I''a), Q is --C(.dbd.NR)--, wherein R.sup.8' is H, --OH, --CN or
C.sub.1-C.sub.6alkyl. For example, in some embodiments R.sup.8' is
H. In other embodiments, R.sup.8' is --CN. In other embodiments,
R.sup.8' is --OH.
[0283] In other embodiments, the compound has one of the following
structures (I''b), (I''c), (I''d) or (I''e):
##STR00011##
[0284] In other embodiments, the compound has one of the following
structures (I''f), (I''g), (I''h) or (I''i):
##STR00012##
[0285] In some different embodiments, the compound has one of the
following structures (I''j), (I''k), (I''1) or (I''m):
##STR00013##
[0286] In other various embodiments, the compound has the following
structure (I''):
##STR00014##
wherein A is NH or S.
[0287] For example, in some embodiments, the compound has the
following structure (I'''a):
##STR00015##
wherein:
[0288] represents a double or triple bond;
[0289] Q is --C(.dbd.O)--, --C(.dbd.NR.sup.8')--,
--NR.sup.8C(.dbd.O)--, --S(.dbd.O).sub.2-- or
--NR.sup.8S(.dbd.O).sub.2--;
[0290] R.sup.8 is H, C.sub.1-C.sub.6alkyl or hydroxylalkyl;
[0291] R.sup.8 is H, --OH, --CN or C.sub.1-C.sub.6alkyl; and
[0292] when is a double bond then R.sup.9 and R.sup.10 are each
independently H, cyano, carboxyl, C.sub.1-C.sub.6alkyl,
alkoxycarbonyl, aminylalkyl, alkylaminylalkyl, heteroaryl or
hydroxylalkyl or R.sup.9 and R.sup.10 join to form a carbocyclic or
heterocyclic ring;
[0293] when is a triple bond then R.sup.9 is absent and R.sup.10 is
H, C.sub.1-C.sub.6alkyl, aminylalkyl, alkylaminylalkyl or
hydroxylalkyl; and
[0294] A is NH or S.
[0295] In some of the foregoing embodiments of compound (I'''a), Q
is Q is --C(.dbd.O)--, --NR.sup.8C(.dbd.O)--, --S(.dbd.O).sub.2--
or --NR.sup.8S(.dbd.O).sub.2--.
[0296] In some other of the foregoing embodiments of compound
(I'''a), Q is --C(.dbd.NR)--, wherein R.sup.8' is H, --OH, --CN or
C.sub.1-C.sub.6alkyl. For example, in some embodiments R.sup.8' is
H. In other embodiments, R.sup.8' is --CN. In other embodiments,
R.sup.8' is --OH.
[0297] In other embodiments, the compound has one of the following
structures (I'''b), (I'''c), (I'''d) or (I'''e):
##STR00016##
[0298] In still more embodiments, the compound has one of the
following structures (I'''f), (I'''g), (I'''h) or (I'''i):
##STR00017##
[0299] In certain embodiments of any of the foregoing, at least one
of G.sup.1 or G.sup.2 is N. In other embodiments, at least one of
W, X or Y is N or NR.sup.5. In other embodiments, at least one of
W, X or Y is N and at least one of W, X or Y is CR.sup.6. For
example, in some embodiments two of W, X and Y are N and one of W,
X and Y is CR.sup.6.
[0300] In some embodiments, at least one of W, X or Y is N or
NR.sup.5, wherein R.sup.5 is a bond to L.sup.1. In some other
embodiments, at least one of W, X or Y is N or CR.sup.6, wherein
R.sup.6 is a bond to L.sup.1.
[0301] For example, in some different embodiments, the compound has
one of the following structures:
##STR00018##
wherein
[0302] represents a double or triple bond;
[0303] Q is --C(.dbd.O)--, --C(.dbd.NR)--, --NR.sup.8C(.dbd.O)--,
--S(.dbd.O).sub.2-- or --NR.sup.8S(.dbd.O).sub.2--;
[0304] R.sup.8 is H, C.sub.1-C.sub.6alkyl or hydroxylalkyl;
[0305] R.sup.8 is H, --OH, --CN or C.sub.1-C.sub.6alkyl;
[0306] when is a double bond then R.sup.9 and R.sup.10 are each
independently H, cyano, carboxyl, C.sub.1-C.sub.6alkyl,
alkoxycarbonyl, aminylalkyl, alkylaminylalkyl, heteroaryl or
hydroxylalkyl or R.sup.9 and R.sup.10 join to form a carbocyclic or
heterocyclic ring; and
[0307] when is a triple bond then R.sup.9 is absent and R.sup.10 is
H, C.sub.1-C.sub.6alkyl, aminylalkyl, alkylaminylalkyl or
hydroxylalkyl.
[0308] In some embodiments of the compounds of structures (I'n),
(I'o) or (I'p), R.sup.1 is aryl or heteroaryl and R.sup.2a and
R.sup.2b are independently selected from H and halo, for example in
some further embodiments R.sup.1 is aryl or heteroaryl and R.sup.2a
and R.sup.2b are independently selected from halo, such as chloro
and fluoro. In some embodiments, R.sup.1 is aryl or heteroaryl,
R.sup.2a is chloro and R.sup.2b is fluoro. In other embodiments
R.sup.1 is aryl or heteroaryl, one of R.sup.2a or R.sup.2b is halo,
such as chloro or fluoro, and the other one of R.sup.2a or R.sup.2b
is H. In other embodiments of the foregoing, R.sup.6 is H, cyano,
cyanoalkyl, amino, or C.sub.1-C.sub.6 alkyl.
[0309] In other different embodiments, the bond between W and X Y
and Z are both single bonds. For example, in some embodiments the
compound has one of the following structures (I'''''a) or
(I'''''b):
##STR00019##
wherein:
[0310] represents a double or triple bond;
[0311] Q is --C(.dbd.O)--, --C(.dbd.NR.sup.8')--,
--NR.sup.8C(.dbd.O)--, --S(.dbd.O).sub.2-- or
--NR.sup.8S(.dbd.O).sub.2--;
[0312] R.sup.8 is H, C.sub.1-C.sub.6alkyl or hydroxylalkyl;
[0313] R.sup.8 is H, --OH, --CN or C.sub.1-C.sub.6alkyl;
[0314] when is a double bond then R.sup.9 and R.sup.10 are each
independently H, cyano, carboxyl, C.sub.1-C.sub.6alkyl,
alkoxycarbonyl, aminylalkyl, alkylaminylalkyl, heteroaryl or
hydroxylalkyl or R.sup.9 and R.sup.10 join to form a carbocyclic or
heterocyclic ring; and
[0315] when is a triple bond then R.sup.9 is absent and R.sup.10 is
H, C.sub.1-C.sub.6alkyl, aminylalkyl, alkylaminylalkyl or
hydroxylalkyl.
[0316] In some embodiments of the compounds of structures (I'''''a)
or (I'''''b), R.sup.1 is aryl or heteroaryl and R.sup.2a and
R.sup.2b are independently selected from H and halo, for example in
some further embodiments R.sup.1 is aryl or heteroaryl and R.sup.2a
and R.sup.2b are independently selected from halo, such as chloro
and fluoro. In some embodiments, R.sup.1 is aryl or heteroaryl,
R.sup.2a is chloro and R.sup.2b is fluoro. In other embodiments
R.sup.1 is aryl or heteroaryl, one of R.sup.2a or R.sup.2b is halo,
such as chloro or fluoro, and the other one of R.sup.2a or R.sup.2b
is H. In other embodiments of the foregoing, R.sup.6 is H, cyano,
cyanoalkyl, amino, or C.sub.1-C.sub.6 alkyl.
[0317] In yet more of any of the foregoing embodiments, E has the
following structure:
wherein:
##STR00020##
[0318] Q is --C(.dbd.O)--, --C(.dbd.NR.sup.8')--,
--NR.sup.8C(.dbd.O)--, --S(.dbd.O).sub.2-- or
--NR.sup.8S(.dbd.O).sub.2--;
[0319] R.sup.8 is H, C.sub.1-C.sub.6alkyl or hydroxylalkyl;
[0320] R.sup.8 is H, --OH, --CN or C.sub.1-C.sub.6alkyl; and
[0321] R.sup.9 and R.sup.10 are each independently H, cyano,
C.sub.1-C.sub.6alkyl, aminylalkyl, alkylaminylalkyl, or
hydroxylalkyl or R.sup.9 and R.sup.10 join to form a carbocyclic or
heterocyclic ring.
[0322] In some of the foregoing embodiments, Q is-C(.dbd.O)--,
--NR.sup.8C(.dbd.O)--, --S(.dbd.O).sub.2-- or
--NR.sup.8S(.dbd.O).sub.2--.
[0323] In some other of the foregoing embodiments, Q is
--C(.dbd.NR.sup.8')--, wherein R.sup.8' is H, --OH, --CN or
C.sub.1-C.sub.6 alkyl. For example, in some embodiments R.sup.8' is
H. In other embodiments, R.sup.8' is --CN. In other embodiments,
R.sup.8 is --OH.
[0324] In still other of any of the foregoing embodiments, E has
the following structure:
##STR00021##
wherein:
[0325] Q is --C(.dbd.O)--, --NR.sup.8C(.dbd.O)--,
--S(.dbd.O).sub.2-- or --NR.sup.8S(.dbd.O).sub.2--;
[0326] R.sup.8 is H, C.sub.1-C.sub.6alkyl or hydroxylalkyl; and
[0327] R.sup.10 is H, C.sub.1-C.sub.6alkyl, aminylalkyl,
alkylaminylalkyl or hydroxylalkyl.
[0328] In some embodiments ml is 1. In other embodiments m.sup.1 is
2. In still more embodiments, ml is 3. In different embodiments,
m.sup.2 is 1. In some other embodiments, m.sup.2 is 2. In yet still
more embodiments, m.sup.2 is 3.
[0329] In some other particular embodiments of any of the foregoing
compounds, ml is 1, and m.sup.2 is 1. In other embodiments, ml is 1
and, m.sup.2 is 2. In still other embodiments m.sup.1 is 2, and
m.sup.2 is 2. In more embodiments, ml is 1, and m.sup.2 is 3.
[0330] In any of the foregoing embodiments, G.sup.1 and G.sup.2 are
each independently selected from N and CH. In some embodiments, at
least one of G.sup.1 or G.sup.2 is N. In some embodiments, each of
G.sup.1 and G.sup.2 are N. In some embodiments, each of G.sup.1 and
G.sup.2 are N and m.sup.1 and m.sup.2 are each 2. In some other
embodiments, at least one of G.sup.1 or G.sup.2 is CH. In other
embodiments, each of G.sup.1 and G.sup.2 are CH.
[0331] Without wishing to be bound by theory, Applicants believe
correct selection of the R.sup.1 substituent may play a part in the
compounds' inhibitory activity (e.g., against KRAS, HRAS or NRAS
G12C). In some embodiments, R.sup.1 is aryl or heterocyclyl (e.g.,
heteroaryl or aliphatic heterocyclyl), each of which is optionally
substituted with one or more substituents. In some embodiments,
R.sup.1 is capable of reversible interaction with KRAS, HRAS or
NRAS G12C mutant protein. In some embodiments R.sup.1 has high
affinity towards KRAS, HRAS or NRAS and is highly specific towards
G12C KRAS, HRAS or NRAS. In some embodiments R.sup.1 is capable of
hydrophobic interaction with KRAS, HRAS or NRAS G12C. In some
embodiments R.sup.1 is able to form hydrogen bonds with various
residues of G12C KRAS, HRAS or NRAS protein.
[0332] In other of the foregoing embodiments, R.sup.1 is
heterocyclyl, heteroaryl or aryl.
[0333] In certain embodiments of any of the foregoing, R.sup.1 is
aryl. For example, in some embodiments R.sup.1 is phenyl. In other
embodiments, R.sup.1 is naphthyl. In some of these embodiments,
R.sup.1 is unsubstituted aryl, such as unsubstituted phenyl or
unsubstituted naphthyl. In other embodiments, R.sup.1 is
substituted with one or more substituents. In some of these
embodiments, the substituents are selected from halo, cyano,
hydroxyl, C.sub.1-C.sub.6alkyl, C.sub.1-C.sub.6alkoxy and
C.sub.3-C.sub.8cycloalkyl. In other more specific embodiments, the
substituents are selected from fluoro, chloro, bromo, hydroxyl,
methoxy and cyclopropyl.
[0334] In other embodiments, the R.sup.1 substituents are selected
from halo, cyano, cyanoC.sub.1-C.sub.6alkyl,
cyanoC.sub.3-C.sub.8cycloalkyl, hydroxyl, C.sub.1-C.sub.6alkyl,
C.sub.1-C.sub.6alkylcycloalky, C.sub.2-C.sub.6alkynyl,
C.sub.1-C.sub.6alkoxy, C.sub.1-C.sub.6haloalkoxy,
C.sub.1-C.sub.6alkylaminyl, C.sub.1-C.sub.6alkylcarbonylaminyl,
C.sub.1-C.sub.6 hydroxylalkyl, C.sub.1-C.sub.6haloalkyl,
C.sub.1-C.sub.6alkoxyalkyl, aminylsulfone, aminylcarbonyl,
aminylcarbonylC.sub.1-C.sub.6alkyl,
aminylcarbonylC.sub.3-C.sub.8cycloalkyl,
C.sub.1-C.sub.6alkylaminylcarbonyl,
C.sub.3-C.sub.8cycloalkylaminylcarbonyl,
C.sub.3-C.sub.8cycloalkylalkyl and C.sub.3-C.sub.8cycloalkyl,
C.sub.3-C.sub.8 fusedcycloalkyl and heteroaryl.
[0335] In still other embodiments, the R.sup.1 substituents are
selected from fluoro, chloro, bromo, cyano, hydroxyl,
hydroxylmethyl, methoxy, methoxymethyl, ethyl, isopropyl,
trifluoromethyl, aminylcarbonyl and cyclopropyl.
[0336] In still more embodiments, the R.sup.1 substituents are
selected from fluoro, chloro, bromo, cyano, hydroxyl,
hydroxylmethyl, methoxy, methoxymethyl, methyl, ethyl, isopropyl,
difluoromethyl, trifluoromethyl, aminylcarbonyl and
cyclopropyl.
[0337] In certain embodiments, R.sup.1 has one of the following
structures:
##STR00022## ##STR00023## ##STR00024## ##STR00025##
##STR00026##
[0338] In other of the foregoing embodiments, R.sup.1 has one of
the following structures:
##STR00027## ##STR00028##
[0339] In still other embodiments, R.sup.1 has one of the following
structures:
##STR00029## ##STR00030## ##STR00031## ##STR00032##
##STR00033##
[0340] In some different embodiments of any of the foregoing,
R.sup.1 is heteroaryl. In certain embodiments, R.sup.1 comprises
oxygen, sulfur, nitrogen or combinations thereof. In some of these
embodiments, R.sup.1 comprises sulfur or nitrogen. In certain
embodiments, R.sup.1 is thiophenyl, pyridinyl, pyridinonyl,
pyrimidinyl, benzooxazolyl, benzoisoxazolyl, benzodioxazolyl,
benzoimidazolyl, quinolinyl, quinolinonyl, dihydroquinolinonyl,
tetrahydroquinolinyl, quinazolinyl, indazolyl, .indolinonyl,
benzothiophenyl or dihydrobenzodioxinyl.
[0341] In some embodiments, R.sup.1 is substituted or unsubstituted
indazolyl. In some of these embodiments the indazolyl is
substituted with one or more C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6
alkoxy and/or halo groups. For example, in some embodiments, the
indazolyl is substituted with one or more methyl, methoxy, chloro
and/or fluoro groups.
[0342] For example, in some embodiments R.sup.1 is pyridinyl. In
some embodiments R.sup.1 is unsubstituted pyridinyl, for example
unsubstituted pyridin-4-yl or unsubstituted pyridin-3-yl. In other
embodiments R.sup.1 is thiophenyl. In some embodiments R.sup.1 is
unsubstituted thiophenyl, for example unsubstituted
thiophen-2-yl.
[0343] In other embodiments, R.sup.1 is substituted with one or
more substituents. For example, in some embodiments, the
substituents are selected from halo, C.sub.1-C.sub.6alkyl,
C.sub.1-C.sub.6alkoxy, or C.sub.2-C.sub.6alkenylcarbonylaminyl. In
some of these embodiments, the substituents are selected from halo
and C.sub.1-C.sub.6alkyl. In other embodiments, the substituents
are selected from fluoro, chloro, amino and methyl. For example, in
more specific embodiments, the substituents are selected from
chloro and methyl. In other embodiments at least one R1 substituent
is fluoro.
[0344] In some embodiments, R.sup.1 has one of the following
structures:
##STR00034## ##STR00035## ##STR00036##
[0345] In certain embodiments, R.sup.1 has one of the following
structures:
##STR00037## ##STR00038##
[0346] In some of the foregoing embodiments, R.sup.1 has one of the
following structures:
##STR00039##
[0347] In still other embodiments, R.sup.1 is aliphatic
heterocyclyl. In some embodiments the aliphatic heterocyclyl
comprises oxygen and/or nitrogen. In some further embodiments,
R.sup.1 is morpholinyl. For example, in some embodiments R.sup.1
has the following structure:
##STR00040##
[0348] In various embodiments of the foregoing, R.sup.1 is
unsubstituted.
[0349] In some of the foregoing embodiments, R.sup.2a is H. In
other embodiments, R.sup.2a is halo, for example in some
embodiments R.sup.2a is chloro or fluoro. In still other
embodiments of the foregoing, R.sup.2a is C.sub.1-C.sub.6alkyl. For
example, in some embodiments R.sup.2a is C.sub.3-C.sub.8
cycloalkyl, such as cyclopropyl.
[0350] In other embodiments of the foregoing compounds, R.sup.2b
and R.sup.2c, when present, are H. In different embodiments,
R.sup.2b and R.sup.2c, when present, are each independently halo.
In yet other embodiments, R.sup.2b, when present, is halo. In more
embodiments, R.sup.2c, when present, is halo. In certain of the
foregoing embodiments, halo is chloro or fluoro.
[0351] The Q moiety is typically selected to optimize the
reactivity (i.e., electrophilicity) of E. In certain of the
foregoing embodiments, Q is --C(.dbd.O)--. In other embodiments, Q
is --S(.dbd.O).sub.2--. In still more embodiments, Q is
--NR.sup.8C(.dbd.O)--. In still more different embodiments, Q is
--NR.sup.8S(.dbd.O).sub.2--.
[0352] In some of the immediately foregoing embodiments, R.sup.8 is
H. In other of these embodiments, R.sup.8 is hydroxylalkyl, for
example in some embodiments the hydroxylalkyl is
2-hydroxylalkyl.
[0353] In some embodiments, Q is --C(.dbd.NR)--, wherein R.sup.8'
is H, --OH, --CN or C.sub.1-C.sub.6alkyl. For example, in some
embodiments R.sup.8' is H. In other embodiments, R.sup.8 is
--CN.
[0354] In other embodiments, R.sup.8' is --OH.
[0355] In some of any one of the foregoing embodiments, at least
one of R.sup.9 or R.sup.10 is H. For example, in some embodiments
each of R.sup.9 and R.sup.10 are H.
[0356] In other of the foregoing embodiments, R.sup.10 is
alkylaminylalkyl. In some of these embodiments, R.sup.10 has the
following structure:
##STR00041##
[0357] In other embodiments, R.sup.10 is hydroxylalkyl, such as
2-hydroxylalkyl.
[0358] In some other different embodiments of the foregoing
embodiments, R.sup.9 and R.sup.10 join to form a carbocyclic ring.
For example, in some of these embodiments the carbocyclic ring is a
cyclopentene, cyclohexene or phenyl ring. In other embodiments, the
carbocyclic ring is a cyclopentene or cyclohexene ring. In other
embodiments, the carbocyclic ring is a phenyl ring, for example a
phenyl ring having the following structure:
##STR00042##
[0359] In some of any of the foregoing embodiments E is an
electrophile capable of bonding with a KRAS, HRAS or NRAS protein
comprising G12C mutation. In some embodiments, the electrophile E
is capable of forming an irreversible covalent bond with a G12C
mutant KRAS, HRAS or NRAS protein. In some cases, the electrophile
E may bind with the cysteine residue at the position 12 of a G12C
mutant KRAS, HRAS or NRAS protein. In various embodiments of any of
the foregoing, E has one of the following structures:
##STR00043## ##STR00044##
[0360] In other embodiments of any of the foregoing, E has one of
the following structures:
##STR00045##
[0361] In different embodiments, E has one of the following
structures:
##STR00046##
[0362] In some cases E has one of the following structures:
##STR00047##
wherein:
[0363] R.sup.8 is H or C.sub.1-C.sub.6alkyl;
[0364] R.sup.9 is H, cyano or C.sub.1-C.sub.6alkyl, or R.sup.9
joins with R.sup.10 to form a carbocycle;
[0365] R.sup.10 is H or C.sub.1-C.sub.6alkyl or R.sup.10 joins with
R.sup.9 to form a carbocycle and
[0366] R.sup.10a is H or C.sub.1-C.sub.6alkyl.
[0367] In some embodiments E is
##STR00048##
In some embodiments E is
##STR00049##
In some embodiments E is
##STR00050##
[0368] In some of any of the foregoing embodiments, L.sup.1 is a
bond. In other embodiments, L.sup.1 is NR.sup.7. For example, in
some of these embodiments, R.sup.7 is C.sub.1-C.sub.6alkyl. In
other embodiments, L.sup.1 is NH.
[0369] L.sup.2 can be selected to provide proper spacing and/or
orientation for the E group to form a bond with the KRAS, HRAS or
NRAS protein. In some of the foregoing embodiments, L.sup.2 is a
bond. In other of the foregoing embodiments, L.sup.2 is alkylene.
In some embodiments, the alkylene is substituted. In other
embodiments the alkylene is unsubstituted. For example, in some
embodiments L.sup.2 is CH.sub.2 or CH.sub.2CH.sub.2.
[0370] In certain embodiments, R.sup.3a and R.sup.3b are, at each
occurrence, independently H, --OH, --NH.sub.2, --CO.sub.2H, halo,
cyano, hydroxylalkly, aminylalkyl, cyanoalkyl, carboxyalkyl or
aminylcarbonyl, and R.sup.4a and R.sup.4b are, at each occurrence,
independently H, --OH, --NH.sub.2, --CO.sub.2H, halo, cyano,
hydroxylalkly, aminylalkyl, cyanoalkyl, carboxyalkyl or
aminylcarbonyl.
[0371] In other of the foregoing embodiments, R.sup.3a and R.sup.4a
are, at each occurrence, independently H, --OH, hydroxylalkly,
cyano, or aminylcarbonyl and R.sup.3b and R.sup.4b are H.
[0372] In certain other embodiments, R.sup.3a and R.sup.4a are H
and R.sup.3b and R.sup.4b are, at each occurrence, independently H,
--OH, --NH.sub.2, --CO.sub.2H, halo, cyano, hydroxylalkly,
aminylalkyl, cyanoalkyl, carboxyalkyl or aminylcarbonyl.
[0373] In any of the foregoing embodiments, at least one of
R.sup.3a, R.sup.3b, R.sup.4a or R.sup.4b is H. In some embodiments,
each of R.sup.3a, R.sup.3b, R.sup.4a and R.sup.4b are H.
[0374] In some embodiments, R.sup.3a is --OH, --NH.sub.2,
--CO.sub.2H, halo, cyano, hydroxylalkly, aminylalkyl, cyanoalkyl,
carboxyalkyl or aminylcarbonyl, and R.sup.3b, R.sup.4a and R.sup.4b
are H.
[0375] In other embodiments, R.sup.4a is --OH, --NH.sub.2,
--CO.sub.2H, halo, cyano, hydroxylalkly, aminylalkyl, cyanoalkyl,
carboxyalkyl or aminylcarbonyl, and R.sup.3a, R.sup.3b and R.sup.4b
are H.
[0376] In other embodiments, R.sup.3a is H, --OH, --NH.sub.2,
--CO.sub.2H, halo, cyano, hydroxylalkly, aminylalkyl, cyanoalkyl,
carboxyalkyl or aminylcarbonyl, and R.sup.3b joins with R.sup.4b to
form a carbocyclic or heterocyclic ring; In still more embodiments,
R.sup.4a is H, --OH, --NH.sub.2, --CO.sub.2H, halo, cyano,
hydroxylalkly, aminylalkyl, cyanoalkyl, carboxyalkyl or
aminylcarbonyl, and R.sup.4b joins with R.sup.3b to form a
carbocyclic or heterocyclic ring.
[0377] In other embodiments, R.sup.3a and R.sup.3b join to form a
carbocyclic or heterocyclic ring. In other embodiments, R.sup.4a
and R.sup.4b join to form a carbocyclic or heterocyclic ring.
[0378] In still other embodiments, R.sup.3a or R.sup.4a is
aminylcarbonyl. For example, in certain embodiments, the
aminylcarbonyl is
##STR00051##
In other embodiments, R.sup.3a or R.sup.4a is cyano. In other
embodiments, R.sup.3a or R.sup.4a is --OH. In other embodiments,
R.sup.3a or R.sup.4a is hydroxylalkyl, for example
hydroxylmethyl.
[0379] In some embodiments, R.sup.6 is, at each occurrence,
independently H, oxo, cyano, cyanoalkyl, aminyl, aminylalkyl,
aminylalkylaminyl, aminylcarbonyl, alkylaminyl, haloalkylaminyl,
hydroxylalkyaminyl, amindinylalkyl, amidinylalkoxy,
amindinylalkylaminyl, guanidinylalkyl, guanidinylalkoxy,
guanidinylalkylaminyl, C.sub.1-C.sub.6 alkoxy, aminylalkoxy,
alkylcarbonylaminylalkoxy, C.sub.1-C.sub.6 alkyl, heterocyclyl,
heterocyclyloxy, heterocyclylalkyloxy, heterocyclylaminyl,
heterocyclylalkylaminyl, heteroaryl, heteroaryloxy,
heteroarylalkyloxy, heteroarylaminyl, heteroarylalkylaminyl, aryl,
aryloxy, arylamino, arylalkylamino, arylalkyloxy or a bond to
L.sup.1.
[0380] Each of the foregoing R.sup.6 moieties may be substituted
with one or more substituents. For example, in some embodiments the
one or more substituents are aminyl (e.g., substituted or
substituted), alkylcarbonyl aminyl, hydroxyl, haloalkyl or
heterocycyclyl (e.g., substituted or substituted aliphatic
heterocycle or substituted or substituted heteroaryl). For example,
in some embodiments, the R.sup.6 moiety is C.sub.1-C.sub.6 alkyl,
C.sub.1-C.sub.6 alkoxy or alkylaminyl, which is further substituted
with alkylcarbonylaminyl, hydroxyl, --CN or haloalkyl. For example,
in some embodiments, R.sup.6 has one of the following
structures:
##STR00052##
wherein X is a bond, --O-- or --NR--; each R is independently H or
C.sub.1-C.sub.6alkyl and n is an integer from 0 to 6.
[0381] Various different R.sup.6 moieties are included in the scope
of the compounds. For example, in various embodiments, R.sup.6 is
H. In other embodiments, R.sup.6 is --CN. In more embodiments,
R.sup.6 is methoxy.
[0382] In various other embodiments, R.sup.6 is aminylalkyl,
aminylalkyloxy or aminylalkyaminyl. For example, in some
embodiments R.sup.6 has the following structures:
##STR00053##
wherein X is a bond, --O-- or --NR--; each R is independently H or
C.sub.1-C.sub.6alkyl and n is an integer from 0 to 6.
[0383] In other embodiments, R.sup.6 is amindinylalkyl,
amidinylalkoxy, amindinylalkylaminyl, guanidinylalkyl,
guanidinylalkoxy or guanidinylalkylaminyl. For example, in some
embodiments R.sup.6 has one of the following structures:
##STR00054##
wherein X is a bond, --O-- or --NR--; each R is independently H or
C.sub.1-C.sub.6alkyl and n is an integer from 0 to 6.
[0384] In other embodiments, R.sup.6 is heterocyclyl,
heterocyclyloxy, heterocyclylalkyloxy, heterocyclylaminyl,
heterocyclylalkylaminyl, heteroaryl, heteroaryloxy,
heteroarylalkyloxy, heteroarylaminyl or heteroarylalkylaminyl. For
example, in some embodiments R.sup.6 has one of the following
structures:
##STR00055## ##STR00056##
wherein X is a bond, --O-- or --NR--; each R is independently H or
C.sub.1-C.sub.6alkyl and n is an integer from 0 to 6.
[0385] In some of the foregoing embodiments, X is N. in other of
the foregoing embodiments, X is N. In other of the foregoing
embodiments, Z is N. In still more embodiments, X is N and Z is
N.
[0386] In some embodiments, Z is N and Y is N. In other
embodiments, X is N, Z is N, Y is CR.sup.6, wherein R.sup.6 is H
and W is CR.sup.6, wherein R.sup.6 is a bond to L.sup.1. In
different embodiments, Z is N and Y is CR.sup.6, wherein R.sup.6 is
H, W is CR.sup.6, wherein R.sup.6 is a bond to L.sup.1 and X is
CR.sup.6, wherein R.sup.6 is cyano, methoxy or amino.
[0387] In other embodiments, Z is N, X is CR.sup.6 and R.sup.6 is
cyano, Y is CR.sup.6, wherein R.sup.6 is H and W is CR.sup.6,
wherein R.sup.6 is a bond to L.sup.1.
[0388] In other embodiments, Y is N, Z is N, W is CR.sup.6, wherein
R.sup.6 is a bond to L.sup.1 and X is CR.sup.6, wherein R.sup.6 is
H.
[0389] In other of the foregoing embodiments, Z is a bond.
[0390] In certain embodiments, Y is NR.sup.5. In some of these
embodiments, R.sup.5 is C.sub.1-C.sub.6alkyl. In other embodiments,
R.sup.5 is H.
[0391] In still other embodiments, X or Y is CR.sup.6. In some of
these embodiments, R.sup.6 is, at each occurrence, independently H,
cyano, amino, C.sub.1-C.sub.6alkoxy or a bond to L.sup.1. In some
other of these embodiments, R.sup.6 is H. In other embodiments,
R.sup.6 is C.sub.1-C.sub.6alkoxy. In other embodiments, R.sup.6 is
cyano. In more embodiments, R.sup.6 is methoxy. In other
embodiments, R.sup.6 is amino.
[0392] In various different embodiments, the compound has one of
the structures set forth in Table 1 below:
TABLE-US-00001 TABLE 1 Exemplary Compounds of Structure (I) No.
Structure Name Method [M + H].sup.+ I-1 ##STR00057##
1-(4-(7-chloro-6-(2- chlorophenyl)quinazolin-
4-yl)piperazin-1-yl)prop- 2-en-1-one A 413.20 I-2 ##STR00058##
1-(4-(7-chloro-6-(2- chlorophenyl)quinazolin-
4-ylamino)piperidin-1- yl)prop-2-en-1-one A 427.25 I-3 ##STR00059##
1-(4-(6-chloro-5-(2- chlorophenyl)-1H- indazol-3-yl)piperazin-1-
yl)prop-2-en-1-one C 401.20 I-4 ##STR00060## 1-(4-(7-chloro-6-(4-
chlorophenyl)quinazolin- 4-yl)piperazin-1-yl)prop- 2-en-1-one B
413.25 I-5 ##STR00061## 1-(4-(7-chloro-6-(3-
chlorophenyl)quinazolin- 4-yl)piperazin-1-yl)prop- 2-en-1-one B
413.20 I-6 ##STR00062## 1-(4-(7-chloro-6-(2,4-
dichlorophenyl)quinazolin- 4-yl)piperazin-1- yl)prop-2-en-1-one B
447.20.sup.# I-7 ##STR00063## 1-(4-(7-chloro-6-(3,4-
dichlorophenyl)quinazolin- 4-yl)piperazin-1- yl)prop-2-en-1-one B
449.15 I-8 ##STR00064## 2-(4-(4-acryloylpiperazin-
1-yl)-7-chloroquinazolin- 6-yl)benzonitrile B 404.1 I-9
##STR00065## 1-(4-(7-chloro-6-(2,5- dichlorophenyl)quinazolin-
4-yl)piperazin-1- yl)prop-2-en-1-one B 448.45 I-10 ##STR00066##
1-(4-(7-chloro-6-(5- chloro-2- hydroxyphenyl)quinazolin-
4-yl)piperazin-1- yl)prop-2-en-1-one B 429.25 I-11 ##STR00067##
1-(4-(7-chloro-6-(4- chloro-2- hydroxyphenyl)quinazolin-
4-yl)piperazin-1- yl)prop-2-en-1-one B 429.25 I-12 ##STR00068##
1-(4-(7-chloro-6-(4- hydroxyphenyl)quinazolin- 4-yl)piperazin-1-
yl)prop-2-en-1-one B 395.25 I-13 ##STR00069## 1-(4-(7-chloro-6-(4-
chloro-2- methoxyphenyl)quinazolin- 4-yl)piperazin-1-
yl)prop-2-en-1-one B 443.30 I-14 ##STR00070## 1-(4-(7-chloro-6-(3-
hydroxyphenyl)quinazolin- 4-yl)piperazin-1- yl)prop-2-en-1-one B
395.25 I-15 ##STR00071## 1-(4-(7-chloro-6-(2-
hydroxyphenyl)quinazolin- 4-yl)piperazin-1- yl)prop-2-en-1-one B
395.25 I-16 ##STR00072## 4-(4-(4-acryloylpiperazin-
1-yl)-7-chloroquinazolin- 6-yl)benzonitrile B 404.3 I-17
##STR00073## 1-(4-(7-chloro-6-(pyridin- 4-yl)quinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one B 380.25 I-18 ##STR00074##
1-(4-(7-chloro-6- phenylquinazolin-4- yl)piperazin-1-yl)prop-2-
en-1-one B 379.25 I-19 ##STR00075## 3-(4-(4-acryloylpiperazin-
1-yl)-7-chloroquinazolin- 6-yl)benzonitrile B 404.25 I-20
##STR00076## 1-(4-(7-chloro-6-(pyridin- 3-yl)quinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one B 380.25 I-21 ##STR00077##
1-(4-(7-chloro-6- (thiophen-2- yl)quinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one B 385.25 I-22 ##STR00078##
1-(4-(5-(2-chlorophenyl)- 4a,7a-dihydrothieno[2,3- d]pyrimidin-4-
yl)piperazin-1-yl)prop-2- en-1-one H 385.20 I-23 ##STR00079##
1-(4-(7-chloro-6-(2- chloro-5- fluorophenyl)quinazolin-
4-yl)piperazin-1-yl)prop- 2-en-1-one B 431.20 I-24 ##STR00080##
1-(4-(6-chloro-7-(2- chlorophenyl)isoquinolin-
1-yl)piperazin-1-yl)prop- 2-en-1-one D 412.20 I-25 ##STR00081##
(E)-1-(4-(7-chloro-6-(2- chlorophenyl)quinazolin-
4-yl)piperazin-1-yl)-4- (dimethylamino)but-2-en- 1-one A 470.35
I-26 ##STR00082## 1-(4-(7-chloro-6-(5- methylthiophen-2-
yl)quinazolin-4- yl)piperazin-1-yl)prop-2- en-1-one B 399.20 I-27
##STR00083## 1-(4-(7-chloro-6-(2- chlorophenyl)quinolin-4-
yl)piperazin-1-yl)prop-2- en-1-one E 412.20 I-28 ##STR00084##
1-(4-(5-(2-chlorophenyl)- 7,7a-dihydro-4aH-
pyrrolo[2,3-d]pyrimidin- 4-yl)piperazin-1-yl)prop- 2-en-1-one J
368.25 I-29 ##STR00085## N-(1-(7-chloro-6-(2-
chlorophenyl)quinazolin- 4-yl)azetidin-3- yl)acrylamide B 399.20
I-30 ##STR00086## 1-(3-(7-chloro-6-(2- chlorophenyl)quinazolin-
4-ylamino)azetidin-1- yl)prop-2-en-1-one B 399.20 I-31 ##STR00087##
1-(4-(6-chloro-5-(2- chlorophenyl)-1H- indazol-3-
ylamino)piperidin-1- yl)prop-2-en-1-one C 413.40.sup.+ I-32
##STR00088## 1-(4-(7-chloro-6- morpholinoquinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one L 388.25 I-33 ##STR00089##
1-(4-(6-(2-chlorophenyl)- 7-fluoroquinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one B 397.20 I-34 ##STR00090##
1-(4-(7-chloro-6-(5- chlorothiophen-2- yl)quinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one B 419.15 I-35 ##STR00091##
1-(4-(8-(2- chlorophenyl)quinazolin- 2-yl)piperazin-1-yl)prop-
2-en-1-one I 379.1 I-36 ##STR00092## 1-(4-(7-chloro-6-(2-
chlorophenyl)quinazolin- 4-yl)piperidin-1-yl)prop- 2-en-1-one K
410.35.sup.+ I-37 ##STR00093## 1-(4-(6-chloro-7-(4-
chlorophenyl)isoquinolin- 1-yl)piperazin-1-yl)prop- 2-en-1-one D
412.20 I-38 ##STR00094## 1-(4-(6-chloro-7-(4- chloro-2-
hydroxyphenyl)isoquinolin- 1-yl)piperazin-1- yl)prop-2-en-1-one D
428.25 I-39 ##STR00095## 1-(4-(2-amino-7-chloro- 6-(4-
chlorophenyl)quinazolin- 4-yl)piperazin-1-yl)prop- 2-en-1-one F
428.3 I-40 ##STR00096## 1-(4-(6-(4-bromophenyl)-
7-chloroquinazolin-4- yl)piperazin-1-yl)prop-2- en-1-one B 459.25
I-41 ##STR00097## 1-(4-(7-cyclopropyl-6-(4- cyclopropylphenyl)
quinazolin-4-yl)piperazin-1- yl)prop-2-en-1-one B 425.25 I-42
##STR00098## 4-(4-acryloylpiperazin-1- yl)-7-chloro-6-(4-
chlorophenyl)quinoline- 3-carbonitrile G 437.25 I-43 ##STR00099##
1-(4-(7-chloro-6-(4- chlorophenyl)-2- methoxyquinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one F 465.30* I-44 ##STR00100##
1-acryloyl-4-(7-chloro-6- (4- chlorophenyl)quinazolin-
4-yl)piperazine-2- carboxamide A 454.35.sup.+ I-45 ##STR00101##
7-chloro-6-(4- chlorophenyl)-4-(4- (vinylsulfonyl)piperazin-
1-yl)quinazoline A 449.25 I-46 ##STR00102## 1-(4-(7-chloro-6-(4-
chlorophenyl)quinazolin- 4-yl)-2- (hydroxymethyl)piperazin-
1-yl)prop-2-en-1-one A 443.30 I-47 ##STR00103##
1-acryloyl-4-(7-chloro-6- (4- chlorophenyl)quinazolin-
4-yl)piperazine-2- carbonitrile A 438.25 I-48 ##STR00104##
1-acryloyl-4-(7- chloroquinazolin-4- yl)piperaine-2- carbonitrile A
328.2 I-49 ##STR00105## 1-acryloyl-4-(6-bromo-7-
chloroquinazolin-4- yl)piperazine-2- carbonitrile A 408.20 I-50
##STR00106## 1-(4-(7-chloro-6-(4- chlorophenyl)-2-
methylquinazolin-4- yl)piperazin-1-yl)prop-2- en-1-one M 427.35
I-51 ##STR00107## 1-acryloyl-4-(7-chloro-6- (thiophen-2-
yl)quinazolin-4- yl)piperazine-2- carbonitrile A 410.30 I-52
##STR00108## 1-acryloyl-4-(7-chloro-6- phenylquinazolin-4-
yl)piperazine-2- carbonitrile A 404.35 I-53 ##STR00109##
4-(4-acryloyl-3- cyanopiperazin-1-yl)-7- chloroquinazoline-6-
carbonitrile B 353.20 I-54 ##STR00110## (S)-1-acryloyl-4-(7-
chloro-6-(4- chlorophenyl)quinazolin- 4-yl)piperazine-2-
carboxamide A 456.30 I-55 ##STR00111## 1-acryloyl-4-(7-chloro-6-
cyclopropylquinazolin-4- yl)piperazine-2- carbonitrile B 368.25
I-56 ##STR00112## 1-acryloyl-4-(7-chloro-6- (4-chlorophenyl)-2-
methylquinazolin-4- yl)piperazine-2- carbonitrile M 452.30 I-57
##STR00113## 1-acryloyl-4-(quinazolin- 4-yl)piperazine-2-
carbonitrile A 294.20 I-58 ##STR00114## (R)-1-acryloyl-4-(7-
chloro-6-(4- chlorophenyl)quinazolin- 4-yl)piperazine-2-
carbonitrile A 438.20 I-59 ##STR00115## (S)-1-acryloyl-4-(7-
chloro-6-(4- chlorophenyl)quinazolin- 4-yl)piperazine-2-
carbonitrile A 438.25 I-60 ##STR00116## 1-(4-(7-chloro-6-(4-
chlorophenyl)quinazolin- 4-yl)-2- ((dimethylamino)methyl)
piperazin-1-yl)prop-2-en- 1-one A 470.35 I-61 ##STR00117##
1-acryloyl-4-(6- chloroisoquinolin-1- yl)piperazine-2- carbonitrile
D 327.20 I-62 ##STR00118## 1-(4-(7-chloro-6-(4-
chlorophenyl)quinazolin- 4-yl)-2-(2- hydroxyethyl)piperazin-
1-yl)prop-2-en-1-one A 457.35 I-63 ##STR00119##
(S)-1-(4-(7-chloro-6-(4- chlorophenyl)quinazolin- 4-yl)-2-
(hydroxymethyl)piperazin- 1-yl)prop-2-en-1-one A 443.30 I-64
##STR00120## (R)-1-acryloyl-4-(7- chloro-6-(4-
chlorophenyl)quinazolin- 4-yl)piperazine-2- carboxamide A 456.30
I-65 ##STR00121## (R)-1-(4-(7-chloro-6-(4- chlorophenyl)quinazolin-
4-yl)-2- (hydroxymethyl)piperazin- 1-yl)prop-2-en-1-one A 443.35
I-66 ##STR00122## (E)-4-(7-chloro-6-(4- chlorophenyl)quinazolin-
4-yl)-1-(4- (dimethylamino)but-2- enoyl)piperazine-2- carbonitrile
A 495.40 I-67 ##STR00123## 1-(4-(6-chloro-7- phenylquinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one B 379.30 I-68 ##STR00124##
1-(4-(6-chloro-7- cyclopropylquinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one A 343.25 I-69 ##STR00125##
2-(1-acryloyl-4-(7-chloro- 6-(4- chlorophenyl)quinazolin-
4-yl)piperazin-2- yl)acetamide A 470.35 I-70 ##STR00126##
2-(1-acryloyl-4-(7-chloro- 6-(4- chlorophenyl)quinazolin-
4-yl)piperain-2- yl)acetonitrile A 452.35 I-71 ##STR00127##
1-(4-(6-(4- chlorophenyl)quinazolin- 4-yl)piperazin-1-yl)prop-
2-en-1-one A 379.30 I-72 ##STR00128## 1-(4-(6-chloro-7-(2-
chlorophenyl)quinazolin- 4-yl)piperazin-1-yl)prop- 2-en-1-one A
413.25 I-73 ##STR00129## 1-(4-(6-chloro-7-(3-
chlorophenyl)quinazolin- 4-yl)piperazin-1-yl)prop- 2-en-1-one A
413.3 I-74 ##STR00130## 1-(4-(6-chloro-7-(2-
hydroxyphenyl)quinazolin- 4-yl)piperazin-1- yl)prop-2-en-1-one A
395.25 I-75 ##STR00131## 1-(4-(6-chloro-7-(3-
hydroxyphenyl)quinazolin- 4-yl)piperazin-1- yl)prop-2-en-1-one A
395.25 I-76 ##STR00132## 1-(4-(6-chloro-7- phenoxyquinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one L 395.25 I-77 ##STR00133##
1-(4-(6-chloro-7-(2- ethylphenyl)quinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one A 407.75 I-78 ##STR00134##
1-(4-(6-chloro-7-(4- chlorophenyl)quinazolin-
4-yl)piperazin-1-yl)prop- 2-en-1-one A 413.25 I-79 ##STR00135##
1-(4-(6-chloro-7-(3- ethylphenyl)quinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one A 407.30 I-80 ##STR00136##
1-(4-(6-chloro-7- (piperidin-1- yl)quinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one L 387.25 I-81 ##STR00137##
1-(4-(6-chloro-7-(2- fluorophenyl)quinazolin-
4-yl)piperazin-1-yl)prop- 2-en-1-one A 397.25 I-82 ##STR00138##
(E)-1-(4-(6-chloro-7- phenylquinazolin-4-
yl)piperazin-1-yl)-4- (dimethylamino)but-2-en- 1-one A 436.40 I-83
##STR00139## 1-(4-(6-chloro-7-(4- fluorophenyl)quinazolin-
4-yl)piperazin-1-yl)prop- 2-en-1-one A 397.25 I-84 ##STR00140##
1-(4-(6-chloro-7-(3- fluorophenyl)quinazolin-
4-yl)piperazin-1-yl)prop- 2-en-1-one A 397.25 I-85 ##STR00141##
2-(1-acryloyl-4-(6-chloro- 7-phenylquinazolin-4- yl)piperazin-2-
yl)acetonitrile A 418.30 I-86 ##STR00142## 1-(4-(6-cyclopropyl-7-
phenylquinazolin-4- yl)piperazin-1-yl)prop-2- en-1-one B 385.75
I-87 ##STR00143## 1-(4-(7-phenylquinazolin-
4-yl)piperazin-1-yl)prop- 2-en-1-one B 345.20 I-88 ##STR00144##
1-(4-(7-chloro-6- phenylisoquinolin-1- yl)piperazin-1-yl)prop-2-
en-1-one D 378.20 I-89 ##STR00145## N-(1-(6-chloro-7-
phenylquinazolin-4- yl)piperidin-4- yl)acrylamide B 393.25 I-90
##STR00146## 1-(4-(6-chloro-7-(pyridin- 3-yl)quinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one B 380.25 I-91 ##STR00147##
1-(4-(6-chloro-7- phenylquinolin-4- yl)piperazin-1-yl)prop-2-
en-1-one E 378.20 I-92 ##STR00148## 1-(4-(6-chloro-7-(pyridin-
2-yl)quinazolin-4- yl)piperazin-1-yl)prop-2- en-1-one B 380.25 I-93
##STR00149## 1-(4-(6-ethyl-7- phenylquinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one B 373.75 I-94 ##STR00150##
1-(4-(6-chloro-2- methoxy-7- phenylquinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one F 409.30 I-95 ##STR00151##
1-(4-(6-chloro-2-methyl- 7-phenylquinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one M 393.70 I-96 ##STR00152##
1-(3-(6-chloro-7- phenylquinazolin-4- ylamino)azetidin-1-
yl)prop-2-en-1-one A 365.20 I-97 ##STR00153## 1-(4-(6-chloro-7-(2-
methoxyphenyl)quinazolin- 4-yl)piperazin-1- yl)prop-2-en-1-one B
409.7 I-98 ##STR00154## 2-(4-(4-acryloylpiperazin-
1-yl)-6-chloroquinazolin- 7-yl)benzamide B 422.30 I-99 ##STR00155##
1-(4-(6-chloro-7-(2- isopropylphenyl)quinazolin- 4-yl)piperazin-1-
yl)prop-2-en-1-one B 421.35 I-100 ##STR00156## 1-(4-(6-chloro-7-(2-
(trifluoromethyl)phenyl) quinazolin-4-yl)piperazin-
1-yl)prop-2-en-1-one B 447.80 I-101 ##STR00157##
1-(4-(6-chloro-7-(2,5- dichlorophenyl)quinazolin- 4-yl)piperazin-1-
yl)prop-2-en-1-one B 447.25 I-102 ##STR00158##
1-(4-(6-chloro-7-(2,4- dichlorophenyl)quinazolin- 4-yl)piperazin-1-
yl)prop-2-en-1-one B 447.30 I-103 ##STR00159## 1-(4-(6-chloro-7-(2-
(methoxymethyl)phenyl) quinazolin-4-yl)piperazin-
1-yl)prop-2-en-1-one B 423.35 I-104 ##STR00160##
1-acryloyl-4-(6-chloro-7- phenylquinazolin-4- yl)piperazine-2-
carboxamide B 422.35 I-105 ##STR00161## 2-(4-(4-acryloylpiperazin-
1-yl)-6-chloroquinazolin- 7-yl)benzonitrile B 405.20 I-106
##STR00162## 2-(1-acryloyl-4-(6-chloro- 7-(2-
fluoorophenyl)quinazolin- 4-yl)piperazin-2- yl)acetonitrile B
437.30 I-107 ##STR00163## 2-(1-acryloyl-4-(6-chloro- 7-(2-
ethylphenyl)quinazolin-4- yl)piperazin-2- yl)acetonitrile B 446.35
I-108 ##STR00164## 1-(4-(6-chloro-7-(2- (hydroxymethyl)phenyl)
quinazolin-4-yl)piperazin- 1-yl)prop-2-en-1-one B 409.30 I-109
##STR00165## 2-(1-acryloyl-4-(6-chloro- 7-(2-
chlorophenyl)quinazolin- 4-yl)piperazin-2- yl)acetonitrile B 452.30
I-110 ##STR00166## 2-(1-acryloyl-4-(6-chloro- 7-(4-
chlorophenyl)quinazolin- 4-yl)piperazin-2- yl)acetonitrile B 452.25
I-111 ##STR00167## 2-(1-acryloyl-4-(6-chloro- 7-(4-
chlorophenyl)quinazolin- 4-yl)piperazin-2- yl)acetonitrile B 452.25
I-112 ##STR00168## 1-(4-(6-chloro-7-(2,4-
difluorophenyl)quinazolin- 4-yl)piperazin-1-yl)prop- 2-en-1-one B
415.0 I-113 ##STR00169## 1-(4-(6-chloro-7-(2,5-
difluorophenyl)quinazolin- 4-yl)piperazin-1-yl)prop- 2-en-1-one B
415.10 I-114 ##STR00170## 1-(4-(6-chloro-7-(4- chloro-2-
fluorophenyl)quinazolin- 4-yl)piperazin-1-yl)prop- 2-en-1-one B
431.05 I-115 ##STR00171## 1-(4-(6-chloro-7-(5- chloro-2-
fluorophenyl)quinazolin- 4-yl)piperazin-1-yl)prop- 2-en-1-one B
431.05 I-116 ##STR00172## 1-(4-(6-chloro-7-
phenylquinazolin-4-yl)-2- (hydroxymethyl)piperazin-
1-yl)prop-2-en-1-one B 409.25 I-117 ##STR00173##
1-(4-(6-chloro-7-(4- chloro-2- hydroxyphenyl)quinazolin-
4-yl)piperazin-1- yl)prop-2-en-1-one B 429.35 I-118 ##STR00174##
1-(4-(6-chloro-7-(5- chloro-2- hydroxyphenyl)quinazolin-
4-yl)piperain-1- yl)prop-2-en-1-one B 429.30 I-119 ##STR00175##
1-(4-(6-chloro-7-(4- fluoro-2- (trifluoromethyl)phenyl)
quinazolin-4-yl)piperazin- 1-yl)prop-2-en-1-one B 465.35 I-120
##STR00176## 1-acryloyl-4-(6-chloro-7- (2- fluorophenyl)quinazolin-
4-yl)piperazine-2- carboxamide B 440.30 I-121 ##STR00177##
1-acryloyl-4-(6-chloro-7- (2- (trifluoromethyl)phenyl)
quinazolin-4-yl)piperazine- 2-carboxamide B 490.40 I-122
##STR00178## 1-(4-(6-chloro-7-(5- fluoro-2-
hydroxyphenyl)quinazolin- 4-yl)piperazin-1- yl)prop-2-en-1-one B
413.30 I-123 ##STR00179## 1-(4-(6-chloro-7- (naphthalen-1-
yl)quinazolin-4- yl)piperazin-1-yl)prop-2- en-1-one B 429.35 I-124
##STR00180## 1-(4-(6-chloro-7-(2- (trifluoromethyl)phenyl)
quinazolin-4-yl)-2- methylpiperazin-1- yl)prop-2-en-1-one B 461.35
I-125 ##STR00181## 2-(1-acryloyl-4-(6-chloro- 7-(2-
(trifluoromethyl)phenyl) quinazolin-4-yl)piperazin-
2-yl)acetonitrile B 486.40 I-126 ##STR00182## 1-(4-(6-chloro-7-(2-
cyclopropylphenyl) quinazolin-4-yl)piperazin- 1-yl)prop-2-en-1-one
B 419.20 I-127 ##STR00183## 4-(4-acryloylpiperazin-1-
yl)-6-chloro-7-(2- fluorophenyl)quinoline-3- carbonitrile G 421.30
I-128 ##STR00184## 1-(4-(6-chloro-7-(2- chloro-5-
hydroxyphenyl)quinazolin- 4-yl)piperazin-1- yl)prop-2-en-1-one B
430.10 I-129 ##STR00185## 1-(4-(7-(benzo[d]oxazol-
7-yl)-6-chloroquinazolin- 4-yl)piperazin-1-yl)prop- 2-en-1-one B
420.10 I-130 ##STR00186## 3-(4-(4-acryloylpiperazin-
1-yl)-6-chloroquinazolin- 7-yl)benzonitrile B 404.10 I-131
##STR00187## 3-(4-(4-acryloylpiperazin- 1-yl)-6-chloroquinazolin-
7-yl)-2-fluoro-N,N- dimethylbenzamide B 468.10 I-132 ##STR00188##
1-(4-(6-chloro-7-(2,6- difluorophenyl)quinazolin-
4-yl)piperazin-1-yl)prop- 2-en-1-one B 415.3 I-133 ##STR00189##
1-(4-(6-chloro-7-(4- fluoro-2- hydroxyphenyl)quinazolin-
4-yl)piperazin-1- yl)prop-2-en-1-one B 413.30 I-134 ##STR00190##
1-(4-(6-chloro-7-(2- hydroxyphenyl)quinazolin- 4-yl)-2-
methylpiperazin-1- yl)prop-2-en-1-one B 409.30 I-135 ##STR00191##
1-(4-(6-chloro-7- (quinolin-5-yl)quinazolin-
4-yl)piperazin-1-yl)prop- 2-en-1-one B 430.30 I-136 ##STR00192##
1-(4-(6-chloro-7- (isoquinolin-5- yl)quinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one B 430.35 I-137 ##STR00193##
4-(4-acryloylpiperazin-1- yl)-7-(2- fluorophenyl)quinazoline-
6-carbonitrile B 388.30 I-138 ##STR00194## 1-(4-(6-chloro-7-(2-
fluoro-6- hydroxyphenyl)quinazolin- 4-yl)piperazin-1-
yl)prop-2-en-1-one B 413.25 I-139 ##STR00195##
2-(1-acryloyl-4-(6-chloro- 7-(2,4- difluorophenyl)quinazolin-
4-yl)piperazin-2- yl)acetonitrile B 454.30 I-140 ##STR00196##
1-(4-(6-chloro-7-(5- methyl-1H-indazol-4- yl)quinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one Q 433.15 I-141 ##STR00197##
1-(4-(6-chloro-7-(2- fluoro-5- (trifluoromethoxy)phenyl)
quinazolin-4- yl)piperazin-1-yl)prop-2- en-1-one B 481.10 I-142
##STR00198## 3-(4-(4-acryloylpiperazin- 1-yl)-6-chloroquinazolin-
7-yl)-N- cyclopropylbenzamide B 462.20 I-143 ##STR00199##
1-(3-(4-(4- acryloylpiperazin-1-yl)-6- chloroquinazolin-7-yl)-4-
fluorophenyl) cyclopropanecarbonitrile B 462.10 I-144 ##STR00200##
1-(4-(6-chloro-7-(1H- indazol-5-yl)quinazolin-
4-yl)piperazin-1-yl)prop- 2-en-1-one B 419.25 I-145 ##STR00201##
1-acryloyl-4-(6-chloro-7- (2,4- difluorophenyl)quinazolin-
4-yl)piperazine-2- carbonitrile B 440.30 I-146 ##STR00202##
1-acryloyl-4-(6-chloro-7- (2- hydroxyphenyl)
quinazolin-4-yl)piperazine-2- carbonitrile B 420.25 I-147
##STR00203## 1-(4-(6-chloro-7-(5- cyclopropyl-2-
fluorophenyl)quinazolin- 4-yl)piperazin-1-yl)prop- 2-en-1-one B
437.10 I-148 ##STR00204## 1-(4-(6-chloro-7-(5,6,7,8-
tetrahydronaphthalen-1- yl)quinazolin-4- yl)piperazin-1-yl)prop-2-
en-1-one B 433.20 I-149 ##STR00205## 1-(4-(7-(3-
aminobenzo[d]isoxazol- 4-yl)-6-chloroquinazolin-
4-yl)piperazin-1-yl)prop- 2-en-1-one B 435.30 I-150 ##STR00206##
1-(4-(7-(2-fluorophenyl)- 6- (trifluoromethyl)quinazolin-
4-yl)piperazin-1- yl)prop-2-en-1-one R 431.30 I-151 ##STR00207##
1-(4-acryloylpiperazin-1- yl)-7-chloro-6-(2,4-
difluorophenyl)quinoxalin- 2(1H)-one S 430.30 I-152 ##STR00208##
1-(4-(6-chloro-7-(1H- indazol-7-yl)quinazolin-
4-yl)piperazin-1-yl)prop- 2-en-1-one B 419.30 I-153 ##STR00209##
1-(4-(6-chloro-7-(2- hydroxynaphthalen-1- yl)quinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one Q 445.10 I-154 ##STR00210##
1-(4-(6-chloro-7-(2- ethynylphenyl))quinazolin-
4-yl)piperazin-1-yl)prop- 2-en-1-one B 403.25 I-155 ##STR00211##
3-(4-(4-acryloylpiperazin- 1-yl)-6-chloroquinazolin-
7-yl)-4-fluorobenzamide B 440.25 I-156 ##STR00212##
1-(4-(6-chloro-7-(2- (cyclopropylmethyl)phenyl) quinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one B 433.35 I-157 ##STR00213##
1-(4-(7-(2- (trifluoromethyl)phenyl) quinazolin-4-yl)piperazin-
1-yl)prop-2-en-1-one B 413.10 I-158 ##STR00214##
1-(4-(6-chloro-8-fluoro-7- (2- fluorophenyl)quinazolin-
4-yl)piperazin-1-yl)prop- 2-en-1-one O 415.25 I-159 ##STR00215##
1-(4-(6-chloro-7-(2- fluorophenyl)cinnolin-4-
yl)piperazin-1-yl)prop-2- en-1-one N 397.25 I-160 ##STR00216##
4-(4-(4-acryloylpiperazin- 1-yl)-6-chloroquinazolin-
7-yl)indolin-2-one B 434.25 I-161 ##STR00217## 2-(2-(4-(4-
acryloylpiperazin-1-yl)-6- chloroquinazolin-7- yl)phenyl)acetamide
B 436.1 I-162 ##STR00218## 1-(4-(6-chloro-7-(1H-
indazol-6-yl)quinazolin- 4-yl)piperazin-1-yl)prop- 2-en-1-one B
419.3 I-163 ##STR00219## 1-(4-(7-(2-fluorophenyl)-
6-hydroxyquinazolin-4- yl)piperazin-1-yl)prop-2- en-1-one A
379.25
I-164 ##STR00220## 1-(4-(7-(2- aminobenzo[d]oxazol-5-
yl)-6-chloroquinazolin-4- yl)piperazin-1-yl)prop-2- en-1-one B
435.25 I-165 ##STR00221## 1-(4-(7-(1H- benzo[d]imidazol-4-yl)-6-
chloroquinazolin-4- yl)piperazin-1-yl)prop-2- en-1-one B 419.30
I-166 ##STR00222## 1-(4-(6-(2- (trifluoromethyl)phenyl)
thieno[3,2-d]pyrimidin-4- yl)piperazin-1-yl)prop-2- en-1-one H
419.10 I-167 ##STR00223## 1-(4-(6-chloro-7-(1H-
indazol-4-yl)quinazolin- 4-yl)piperazin-1-yl)prop- 2-en-1-one B
419.30 I-168 ##STR00224## 2-(2-(4-(4- acryloylpiperazin-1-yl)-6-
chloroquinazolin-7- yl)phenyl)acetonitrile B 418.1 I-169
##STR00225## 1-(4-(6-chloro-7-(4- hydroxy-2-
(trifluoromethyl)phenyl) quinazolin-4-yl)piperazin-
1-yl)prop-2-en-1-one B 463.30 I-170 ##STR00226##
3-(4-(4-acryloylpiperazin- 1-yl)-6-chloroquinazolin-
7-yl)pyridin-2(1H)-one B 396.25 I-171 ##STR00227##
4-(4-acryloylpiperazin-1- yl)-6-chloro-7- (naphthalen-1-
yl)quinoline-3- carbonitrile P 453.30 I-172 ##STR00228##
4-(4-acryloylpiperazin-1- yl)-6-chloro-7-(2,4-
difluorophenyl)quinoline- 3-carbonitrile P 439.25 I-173
##STR00229## 4-(4-acryloylpiperazin-1- yl)-6-chloro-7-(2-
(trifluoromethyl)phenyl) quinoline-3-carbonitrile P 471.35 I-174
##STR00230## N-(3-(4-(4- acryloylpiperazin-1-yl)-6-
chloroquinazolin-7-yl)-4- fluorophenyl)acetamide B 454.10 I-175
##STR00231## 1-(2-(4-(4- acryloylpiperazin-1-yl)-6-
chloroquinazollin-7- yl)phenyl) cyclopropanecarbonitrile B 444.1
I-176 ##STR00232## 1-(2-(4-(4- acryloylpiperazin-1-yl)-6-
chloroquinazolin-7- yl)phenyl) cyclopropanecarboxamide B 462.2
I-177 ##STR00233## 1-(4-(4-acryloylpiperazin-
1-yl)-6-chloroquinazolin- 7-yl)-5-chloropyridin- 2(1H)-one T 430.20
I-178 ##STR00234## N-(4-(4-(4- acryloylpiperazin-1-yl)-6-
chloroquinazolin-7-yl)-5- methylpyrimidin-2- yl)acrylamide B 464.10
I-179 ##STR00235## 1-(4-(7-(2-amino-5- methylpyrimidin-4-yl)-6-
chloroquinazolin-4- yl)piperazin-1-yl)prop-2- en-1-one B 410.10
I-180 ##STR00236## 1-(4-(6-chloro-7,8'- biquinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one B 431.10 I-181 ##STR00237##
1-(4-(4-acryloylpiperazin- 1-yl)-6-chloroquinazolin-
7-yl)-4-chloropyridin- 2(1H)-one T 430.10 I-182 ##STR00238##
4-(4-acryloylpiperazin-1- yl)-6-chloro-7-(2-
hydroxyphenyl)quinoline- 3-carbonitrile P 419.15 I-183 ##STR00239##
1-(4-(7-(2-(1H-pyrazol-4- yl)phenyl)-6- chloroquinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one B 445.20 I-184 ##STR00240##
4-(4-acryloylpiperazin-1- yl)-6-chloro-7-(2-chloro- 5-
hydroxyphenyl)quinoline- 3-carbonitrile P 453.15 I-185 ##STR00241##
1-(4-(6-chloro-7- (thiophen-2- yl)quinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one B 385.10 I-186 ##STR00242##
1-(4-(6-chloro-7-(2- (thiazol-2- yl)phenyl)quinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one U 462.25 I-187 ##STR00243##
1-(4-(6-chloro-7-(2- (thiazol-5- yl)phenyl)quinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one U 462.25 I-188 ##STR00244##
1-(4-(6-chloro-7-(2- fluoro-5-(1H-pyrazol-4-
yl)phenyl)quinazolin-4- yl)piperazin-1-yl)prop-2- en-1-one B 463.20
I-189 ##STR00245## 4-(4-acryloylpiperazin-1- yl)-6-chloro-7-(2-
fluorophenyl)quinoline-3- carboxamide P 439.60 I-190 ##STR00246##
1-(4-(7-(2-amino-4- methylpyrimidin-5-yl)-6- chloroquinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one B 410.10 I-191 ##STR00247##
1-(4-(6-chloro-7-(2- methyl-5- (methylamino)phenyl)
quinazolin-4-yl)piperazin-1- yl)prop-2-en-1-one B 422.20 I-192
##STR00248## 2-(4-(4-acryloylpiperazin- 1-yl)-6-chloroquinazolin-
7-yl)-3-fluorobenzonitrile B 422.10 I-193 ##STR00249##
2-(4-(4-acryloylpiperazin- 1-yl)-6-chloroquinazolin-
7-yl)-5-fluorobenzamide B 440.20 I-194 ##STR00250##
1-(4-(6-chloro-7-(2- fluoro-6- methoxyphenyl)quinazolin-
4-yl)piperazin-1- yl)prop-2-en-1-one B 427.15 I-195 ##STR00251##
1-(4-(6-chloro-7-(2,4- difluorophenyl)quinazolin-
4-yl)-2-ethynylpiperazin- 1-yl)prop-2-en-1-one B 439.15 I-196
##STR00252## 4-(4-acryloylpiperazin-1- yl)-6-chloro-7-(2-fluoro- 5-
hydroxyphenyl)quinoline- 3-carbonitrile P 437.15 I-197 ##STR00253##
2-(4-(4-acryloylpiperazin- 1-yl)-6-chloroquinazolin-
7-yl)-4-fluorobenzamide B 440.20 I-198 ##STR00254## 1-(4-(7-
(benzo[b]thiophen-3-yl)- 6-chloroquinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one B 435.15 I-199 ##STR00255##
1-(4-(6-chloro-7-(2,3- difluoro-6- methoxyphenyl)quinazolin-
4-yl)piperazin-1- yl)prop-2-en-1-one B 445.1 I-200 ##STR00256##
1-(4-(6-chloro-7-(2,2- difluorobenzo[d][1,3]dioxol-
4-yl)quinazolin-4- yl)piperazin-1-yl)prop-2- en-1-one B 459.10
I-201 ##STR00257## 1-(4-(6-chloro-7-(2,3-
dihydrobenzo[b][1,4]dioxin- 5-yl)quinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one B 437.1 I-202 ##STR00258##
1-(4-(6-chloro-7-(2- methoxynaphthalen-1- yl)quinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one O 459.15 I-203 ##STR00259##
1-(4-(6-chloro-7-(2,3- difluoro-6- hydroxyphenyl)quinazolin-
4-yl)piperazin-1- yl)prop-2-en-1-one B 431.10 I-204 ##STR00260##
1-(4-(7-(2,4- difluorophenyl)-6- (trifluoromethyl)quinazolin-
4-yl)piperazin-1- yl)prop-2-en-1-one B 449.15 I-205 ##STR00261##
5-(4-(4-acryloylpiperazin- 1-yl)-6-chloroquinazolin- 7-yl)-3,4-
dihydroquinolin-2(1H)- one B 448.15 I-206 ##STR00262##
1-(4-(6-chloro-7-(2,4- difluoro-5- hydroxyphenyl)quinazolin-
4-yl)piperazin-1- yl)prop-2-en-1-one U 431.10 I-207 ##STR00263##
1-(4-(7-(2-chloro-5- hydroxyphenyl)-6-
(trilfluoromethyl)quinazolin- 4-yl)piperazin-1- yl)prop-2-en-1-one
R 463.15 I-208 ##STR00264## 1-(4-(7-(2-fluoro-6- hydroxyphenyl)-6-
(trifluoromethyl)quinazolin- 4-yl)piperazin-1- yl)prop-2-en-1-one R
447.20 I-209 ##STR00265## 1-(4-(6-chloro-8-fluoro-7- (2-
(trifluoromethyl)phenyl) quinazolin-4-yl)piperazin-
1-yl)prop-2-en-1-one O 465.15 I-210 ##STR00266##
1-(4-(6,8-dichloro-7-(2- fluorophenyl)quinazolin-
4-yl)piperazin-1-yl)prop- 2-en-1-one V 431.10 I-211 ##STR00267##
2-(4-(4-acryloylpiperazin- 1-yl)-6- (trifluoromethyl)quinazolin-
7-yl)benzamide R 456.15 I-212 ##STR00268##
1-(4-(6-(trifluoromethyl)- 7-(2- (trifluoromethyl)phenyl)
quinazolin-4-yl)piperazin- 1-yl)prop-2-en-1-one R 481.20 I-213
##STR00269## 2-(4-(4-acryloylpiperazin- 1-yl)-6-chloroquinazolin-
7-yl)benzenesulfonamide B 458.10 I-214 ##STR00270##
1-(4-(6-chloro-7- (quinolin-4-yl)quinazolin-
4-yl)piperazin-1-yl)prop- 2-en-1-one Q 430.10 I-215 ##STR00271##
1-(4-(6-chloro-3-ethynyl- 7-(2- fluorophenyl)quinolin-4-
yl)piperazin-1-yl)prop-2- en-1-one G 430.10 I-216 ##STR00272##
1-(4-(6-chloro-7-(3,6- difluoro-2- hydroxyphenyl)quinazolin-
4-yl)piperazin-1- yl)prop-2-en-1-one U 431.15 I-217 ##STR00273##
1-(4-(6-chloro-7-(2- chloro-5-hydroxyphenyl)- 8-fluoroquinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one O 447.05 I-218 ##STR00274##
1-(4-(7-(2- hydroxynaphthalen-1-yl)- 6-
(trifluoromethyl)quinazolin- 4-yl)piperazin-1- yl)prop-2-en-1-one R
479.20 I-219 ##STR00275## (E)-1-(4-(6-chloro-7-(2,4-
difluorophenyl)quinazolin- 4-yl)piperazin-1-yl)-4-
(dimethylamino)but-2-en- 1-one O 472.10 I-220 ##STR00276##
4-(4-acryloylpiperazin-1- yl)-6-chloro-7-(2-fluoro- 6-
hydroxyphenyl)quinoline- 3-carbonitrile P 437.15 I-221 ##STR00277##
1-(4-(6-chloro-7-(2,4- difluorophenyl)cinnolin-
4-yl)piperazin-1-yl)prop- 2-en-1-one N 415.10 I-222 ##STR00278##
1-(4-(6-chloro-7-(2-(1- methylcyclopropyl)phenyl) quinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one B 433.20 I-223 ##STR00279##
1-(4-(6-chloro-7-(1,2,3,4- tetrahydroquinolin-5- yl)quinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one B 434.15 I-224 ##STR00280##
1-(4-(6-chloro-7-(2,4- difluorophenyl)-8- fluoroquinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one O 433.10 I-225 ##STR00281##
1-(4-(6-chloro-7-(2- (trifluoromethyl)phenyl)
cinnolin-4-yl)piperazin-1- yl)prop-2-en-1-one N 447.05 I-226
##STR00282## 1-(4-(6-chloro-7-(1- methyl-1H-indazol-3-
yl)quinazolin-4- yl)piperazin-1-yl)prop-2- en-1-one B 433.05 I-227
##STR00283## 1-(4-(6-chloro-8-fluoro-7- (2-fluoro-6-
hydroxyphenyl)quinazolin- 4-yl)piperazin-1- yl)prop-2-en-1-one O
431.05 I-228 ##STR00284## (E)-1-(4-(6-chloro-7-(2-
fluorophenyl)quinazolin- 4-yl)piperazin-1-yl)-4-
(dimethylamino)but-2-en- 1-one B 454.15 I-229 ##STR00285##
(E)-1-(4-(6-chloro-8- fluoro-7-(2- fluorophenyl)quinazolin-
4-yl)piperazin-1-yl)-4- (dimethylamino)but-2-en- 1-one O 472.15
I-230 ##STR00286## (E)-4-(dimethylamino)-1- (4-(8-fluoro-6,7-bis(2-
fluorophenyl)quinazolin- 4-yl)piperazin-1-yl)but-2- en-1-one O
532.25 I-231 ##STR00287## 1-(4-(6-chloro-7-(2- fluoro-6-
hydroxyphenyl)cinnolin- 4-yl)piperazin-1-yl)prop- 2-en-1-one N
413.10 I-232 ##STR00288## 2-(4-(4-acryloylpiperazin-
1-yl)-6-chloroquinazolin- 7-yl)-3-fluorobenzamide Q 440.10 I-233
##STR00289## 1-(4-(6-chloro-7-(2- hydroxy-6-
(trifluoromethyl)phenyl) quinazolin-4-yl)piperazin-
1-yl)prop-2-en-1-one B 463.10 I-234 ##STR00290##
1-(4-(6-chloro-8-fluoro-7- (5-methyl-1H-indazol-4- yl)quinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one O 451.1 I-235 ##STR00291##
1-(4-(6-chloro-8-fluoro-7- (2-hydroxynaphthalen-1- yl)quinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one O 463.10 I-236 ##STR00292##
2-(4-(4-acryloylpiperazin- 1-yl)-6-chloro-8- fluoroquinazolin-7-
yl)benzamide O 440.10 I-237 ##STR00293## 1-(4-(7-(5-methyl-1H-
indazol-4-yl)-6- (trifluoromethyl)quinazolin- 4-yl)piperazin-1-
yl)prop-2-en-1-one R 467.20 I-238 ##STR00294## 1-(4-(6-chloro-7-(5-
methyl-1H-indazol-4- yl)cinnolin-4- yl)piperazin-1-yl)prop-2-
en-1-one N 433.10 I-239 ##STR00295## (E)-ethyl 4-(4-(6-chloro-
7-(2,4- difluorophenyl)quinazolin- 4-yl)piperazin-1-yl)-4-
oxobut-2-enoate O 487.10 I-240 ##STR00296##
8-(4-(4-acryloylpiperazin- 1-yl)-6-chloroquinazolin-
7-yl)quinolin-2(1H)-one U 446.10 I-241 ##STR00297##
(E)-2-(4-(6-chloro-7-(2- fluorophenyl)quinazolin-
4-yl)piperazine-1- carbonyl)-4-methylpent- 2-enenitrile B 464.10
I-242 ##STR00298## 4-(4-acryloylpiperain-1-
yl)-6-chloro-8-fluoro-7- (2- fluorophenyl)quinoline-3- carbonitrile
P 439.10 I-243 ##STR00299## 2-(1-acryloyl-4-(6-chloro-
8-fluoro-7-(2- (trifluoromethyl)phenyl) quinazolin-4-yl)piperazin-
2-yl)acetonitrile O 504.10
I-244 ##STR00300## 1-(4-(6-chloro-7-(5- methoxy-1H-indazol-4-
yl)quinazolin-4- yl)piperazin-1-yl)prop-2- en-1-one B 449.10 I-245
##STR00301## (E)-2-(4-(6-chloro-7-(2- fluorophenyl)quinazolin-
4-yl)piperazine-1- carbonyl)-3-(thiazol-5- yl)acrylonitrile B
505.10 I-246 ##STR00302## 4-(4-acryloylpiperazin-1-
yl)-6-chloro-8-fluoro-7- (2-fluoro-6- hydroxyphenyl)quinoline-
3-carbonitrile P 455.15 I-247 ##STR00303## 1-(4-(6-chloro-7-(2,4-
difluorophenyl)quinazolin- 4-yl)piperazin-1-yl)-4-
hydroxybut-2-yn-1-one O 443.1 I-248 ##STR00304##
(E)-4-(4-(6-chloro-7-(2,4- difluorophenyl)quinazolin-
4-yl)piperazin-1-yl)-4- oxobut-2-enoic acid O 459.05 I-249
##STR00305## 1-(4-(6-chloro-7-(3- methoxynaphthalen-1-
yl)quinazolin-4- yl)piperazin-1-yl)prop-2- en-1-one Q 459.1 I-250
##STR00306## 1-(4-(6-chloro-7-(2- hydroxy-5,6,7,8-
tetrahydronaphthalen-1- yl)quinazolin-4- yl)piperazin-1-yl)prop-2-
en-1-one O 449.10 I-251 ##STR00307## 1-(4-(6-chloro-7-(3-
hydroxynaphthalen-1- yl)quinazolin-4- yl)piperazin-1-yl)prop-2-
en-1-one Q 445.10 I-252 ##STR00308## 1-(4-(6-chloro-8-fluoro-7- (2-
fluorophenyl)quinazolin- 4-yl)-2-ethynylpiperazin-
1-yl)prop-2-en-1-one O 439.10 I-253 ##STR00309##
1-(4-(6-chloro-2-(2- (dimethylamino)ethylamino)-
8-fluoro-7-(5-methyl- 1H-indazol-4- yl)quinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one 537.4 I-254 ##STR00310##
1-(4-(6-chloro-2- ((dimethylamino) methylamino)-8-fluoro-7-(5-
methyl-1H-indazol-4- yl)quinazolin-4- yl)piperazin-1-yl)prop-2-
en-1-one 508.3 I-255 ##STR00311## 1-(4-(6-chloro-7-(5,6-
dimethyl-1H-indazol-7- yl)-8-fluoroquinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one 465.1 I-256 ##STR00312##
1-(4-(6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1- yl)quinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one 493.3 I-257 ##STR00313##
1-(4-(6-chloro-8-fluoro-7- (5-methyl-1H-indazol-4- yl)-2-
(methylamino)quinazolin- 4-yl)piperazin-1-yl)prop- 2-en-1-one 480.2
I-258 ##STR00314## 1-(4-(6-chloro-7-(2- hydroxynaphthalen-1-
yl)cinnolin-4- yl)piperazin-1-yl)prop-2- en-1-one 445.1 I-259
##STR00315## (E)-2-(4-(6-chloro-7-(2- fluorophenyl)quinazolin-
4-yl)piperazine-1- carbonyl)-3-(4- methyloxazol-2- yl)acrylonitrile
503.2 I-260 ##STR00316## 4-(4-acryloylpiperazin-1-
yl)-6-chloro-7-(2- hydroxynaphthalen-1- yl)quinoline-3-
carbonitrile 469.1 I-261 ##STR00317## (E)-2-(4-(6-chloro-7-(2-
fluorophenyl)quinazolin- 4-yl)piperazine-1-
carbonyl)-5-hydroxy-4,4- dimethylpent-2-enenitrile 494.4 I-262
##STR00318## 1-(4-(6-chloro-7-(6- methyl-1H-indazol-4-
yl)quinazolin-4- yl)piperazin-1-yl)prop-2- en-1-one 494.3 I-263
##STR00319## (Z)-4-(4-(6-chloro-7-(2,4- difluorophenyl)quinazolin-
4-yl)piperazin-1-yl)-4- oxobut-2-enenitrile 440.1 I-264
##STR00320## 1-(4-(6-chloro-7-(5- chloro-1H-indazol-7-
yl)quinazolin-4- yl)piperazin-1-yl)prop-2- en-1-one 454.1 I-265
##STR00321## 2-(4-(4-acryloylpiperazin- 1-yl)-6-chloroquinazolin-
7-yl)-3- hydroxybenzonitrile 420.1 I-266 ##STR00322##
1-(4-(6-chloro-7-(5- chloro-1H-indazol-4- yl)quinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one 453.1 I-267 ##STR00323##
1-(4-(6-chloro-7-(2- fluoro-5-(2- hydroxypropan-2-
yl)phenyl)quinazolin-4- yl)piperazin-1-yl)prop-2- en-1-one 455.1
I-268 ##STR00324## 1-(4-(6-chloro-7-(6- methyl-1H-indazol-7-
yl)quinazolin-4- yl)piperazin-1-yl)prop-2- en-1-one 433.2 I-269
##STR00325## 4-(4-acryloylpiperazin-1- yl)-6-chloro-8-fluoro-7-
(2-hydroxynaphthalen-1- yl)quinoline-3- carbonitrile 487.1 I-270
##STR00326## 4-(4-acryloylpiperazin-1- yl)-6-chloro-7-(5-methyl-
1H-indazol-4- yl)quinoline-3- carbonitrile 457.1 I-271 ##STR00327##
1-(4-(8-fluoro-7-(2- fluorophenyl)-6- (trifluoromethyl)quinazolin-
4-yl)piperazin-1- yl)prop-2-en-1-one 449.2 I-272 ##STR00328##
1-(4-(6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1- yl)quinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one 463.2 I-273 ##STR00329##
1-(4-(8-fluoro-7-(2- fluoro-6-hydroxyphenyl)- 6-
(trifluoromethyl)quinazolin- 4-yl)piperazin-1- yl)prop-2-en-1-one
465.2 I-274 ##STR00330## 1-(4-(6-chloro-8-fluoro-7-
(6-methyl-1H-indazol-7- yl)quinazolin-4- yl)piperazin-1-yl)prop-2-
en-1-one 451.2 I-275 ##STR00331## 1-(4-(6-chloro-8-fluoro-7-
(4-fluoro-2- (trifluoromethyl)phenyl) quinazolin-4-yl)piperazin-
1-yl)prop-2-en-1-one 483.2 I-276 ##STR00332##
1-(4-(7-(3-(1H-pyrazol-5- yl)phenyl)-6- chloroquinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one 445.2 I-277 ##STR00333##
1-(4-(6-chloro-7-(3,6- difluoro-2- hydroxyphenyl)-8-
fluoroquinazolin-4- yl)piperazin-1-yl)prop-2- en-1-one 449.1 I-278
##STR00334## 1-(4-(6-chloro-8-fluoro-7- (2-(2-hydroxypropan-2-
yl)phenyl)quinazolin-4- yl)piperazin-1-yl)prop-2- en-1-one 455.2
I-279 ##STR00335## 1-(4-(7-(2-fluoro-6- hydroxyphenyl)-6-
(trifluoromethyl)cinnolin- 4-yl)piperazin-1-yl)prop- 2-en-1-one
447.2 I-280 ##STR00336## 1-(4-(6-chloro-7-(2,4- difluoro-6-
hydroxyphenyl)-8- fluoroquinazolin-4- yl)piperazin-1-yl)prop-2-
en-1-one 449.1 I-281 ##STR00337## 1-(4-(6-chloro-8-fluoro-7-
(2-fluoro-5-(1H-imidazol- 4-yl)phenyl)quinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one 481.2 I-282 ##STR00338##
(E)-2-(4-(6-chloro-8- fluoro-7-(2-fluoro-6-
hydroxyphenyl)quinazolin- 4-yl)piperazine-1-
carbonyl)-4-methylpent- 2-enenitrile 498.2 I-283 ##STR00339##
(E)-2-(4-(6-chloro-8- fluoro-7-(2-fluoro-6-
hydroxyphenyl)quinazolin- 4-yl)piperazine-1-
carbonyl)-3-(thiazol-5- yl)acrylonitrile 539.2 I-284 ##STR00340##
(E)-2-(4-(6-chloro-8- fluoro-7-(2-fluoro-6-
hydroxyphenyl)quinazolin- 4-yl)piperazine-1-
carbonyl)-3-(pyridin-2- yl)acrylonitrile 533.2 I-285 ##STR00341##
4-(4-acryloylpiperazin-1- yl)-6-chloro-8-fluoro-7- (2-
(trifluoromethyl)phenyl) quinoline-3-carbonitrile 489.2 I-286
##STR00342## 1-(4-(6,8-dichloro-7-(2- methoxynaphthalen-1-
yl)quinazolin-4- yl)piperazin-1-yl)prop-2- en-1-one 494.1 I-287
##STR00343## 1-(4-(6-chloro-8-fluoro-7- (2-methoxy-6-
methylphenyl)quinazolin- 4-yl)piperazin-1-yl)prop- 2-en-1-one 441.2
I-288 ##STR00344## 1-(4-(6-chloro-8-fluoro-7- (1H-indol-3-
yl)quinazolin-4- yl)piperazin-1-yl)prop-2- en-1-one 436.1 I-289
##STR00345## 1-(4-(6-chloro-7-(2- chloro-6-hydroxyphenyl)-
8-fluoroquinazolin-4- yl)piperazin-1-yl)prop-2- en-1-one 448.0
I-290 ##STR00346## 1-(4-(6-chloro-7-(2- chloro-6-methylphenyl)-
8-fluoroquinazolin-4- yl)piperazin-1-yl)prop-2- en-1-one 427.1
I-291 ##STR00347## 4-(4-acryloylpiperazin-1-
yl)-6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1- yl)quinoline-3-
carbonitrile 487.1 I-292 ##STR00348## 1-(4-(7-(2,4-
difluorophenyl)-8-fluoro- 6-methylquinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one 413.2 I-293 ##STR00349##
4-(4-acryloylpiperazin-1- yl)-6-chloro-8-fluoro-7-
(5-methyl-1H-indazol-4- yl)quinoline-3- carbonitrile 475.2 I-294
##STR00350## 2-(1-acryloyl-4-(6-chloro- 8-fluoro-7-(5-methyl-1H-
indazol-4-yl)quinazolin- 4-yl)piperazin-2- yl)acetonitrile 490.2
I-295 ##STR00351## (E)-1-(4-(6-chloro-8- fluoro-7-(5-methyl-1H-
indazol-4-yl)quinazolin- 4-yl)piperazin-1-yl)-4-
(dimethylamino)but-2-en- 1-one 508.2 I-296 ##STR00352##
1-(4-(7-(2,4- difluorophenyl)-6,8- difluoroquinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one 417.22 I-297 ##STR00353##
1-(4-(6,8-difluoro-7-(5- methyl-1H-indazol-4- yl)quinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one 435.3 I-298 ##STR00354##
1-(4-(6,8-difluoro-7-(6- methyl-1H-indazol-7- yl)quinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one 435.3 I-299 ##STR00355##
1-(4-(6,8-difluoro-7-(2- fluoro-6- hydroxyphenyl)quinazolin-
4-yl)piperazin-1- yl)prop-2-en-1-one 415.3 I-300 ##STR00356##
1-(4-(6-chloro-7-(5- methyl-1H-indazol-4-yl)- 2-(tetrahydrofuran-3-
yloxy)quinazolin-4- yl)piperazin-1-yl)prop-2- en-1-one 519.3 I-301
##STR00357## (E)-1-(4-(6-chloro-8- fluoro-7-(2-fluoro-6-
hydroxyphenyl)quinazolin- 4-yl)piperazin-1-yl)-4-
(dimethylamino)but-2-en- 1-one 488.2 I-302 ##STR00358##
1-(4-(6-chloro-8- methoxy-7-(5-methyl-1H- indazol-4-yl)quinazolin-
4-yl)piperazin-1-yl)prop- 2-en-1-one 463.3 I-303 ##STR00359##
1-(4-(6,8-dichloro-7-(2- hydroxynaphthalen-1- yl)quinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one 479.1 I-304 ##STR00360##
1-(4-(6-chloro-8-fluoro-7- (5-methyl-1H-indazol-4-
yl)-2-(1H-pyrazol-4- yl)quinazolin-4- yl)piperazin-1-yl)prop-2-
en-1-one 517.0 I-305 ##STR00361## 1-(4-(7-(5-methyl-1H-
indazol-4-yl)-6- (trifluoromethyl)cinnolin-
4-yl)piperazin-1-yl)prop- 2-en-1-one 467.2 I-306 ##STR00362##
1-(4-(6-chloro-7-(2,4- difluorophenyl)-8- methoxyquinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one 445.2 I-307 ##STR00363##
1-(4-(6-chloro-7-(5- (difluoromethyl)-2- fluorophenyl)-8-
fluoroquinazolin-4- yl)piperazin-1-yl)prop-2- en-1-one 461.2 I-308
##STR00364## 4-(4-acryloylpiperazin-1- yl)-6-chloro-8-fluoro-7-
(6-methyl-1H-indazol-7- yl)quinoline-3- carbonitrile 475.1 I-309
##STR00365## 1-(4-(6-chloro-8-fluoro-7- (5-methyl-1H-indazol-4-
yl)-2-(1H-pyrazol-5- yl)quinazolin-4- yl)piperazin-1-yl)prop-2-
en-1-one 517.1 I-310 ##STR00366## (E)-6-chloro-4-(4-(4-
(dimethylamino)but-2- enoyl)piperazin-1-yl)-8- fluoro-7-(3-
hydroxynaphthalen-1- yl)quinoline-3- carbonitrile 544.2 I-311
##STR00367## (E)-4-amino-1-(4-(6- chloro-8-fluoro-7-(5-
methyl-1H-indazol-4- yl)quinazolin-4- yl)piperazin-1-yl)but-2-
en-1-one 480.2 I-312 ##STR00368## 1-(4-(6-chloro-7-(5-
methyl-1H-indazol-4-yl)- 2- (methylamino)quinazolin-
4-yl)piperazin-1-yl)prop- 2-en-1-one 462.3 I-313 ##STR00369##
1-(4-(6-chloro-7-(2- fluoro-6-hydroxyphenyl)-
8-methoxyquinazolin-4- yl)piperazin-1-yl)prop-2- en-1-one 443.2
I-314 ##STR00370## 1-(4-(6-chloro-2-(2- (dimethylamino)ethoxy)-
7-(5-methyl-1H-indazol- 4-yl)quinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one 520.4 I-315 ##STR00371##
(E)-1-(4-(6-chloro-8- fluoro-7-(5-methyl-1H-
indazol-4-yl)quinazoin- 4-yl)piperazin-1-yl)-4-
hydroxybut-2-en-1-one 481.3 I-316 ##STR00372## 1-(4-(6-chloro-7-(5-
methyl-1H-indazol-4-yl)- 2-(tetrahydro-2H-pyran-
3-yloxy)quinazolin-4- yl)piperazin-1-yl)prop-2- en-1-one 533.3
I-317 ##STR00373## (E)-6-chloro-4-(4-(4- (dimethylamino)but-2-
enoyl)piperazin-1-yl)-8- fluoro-7-(5-methyl-1H-
indazol-4-yl)quinoline-3- carbonitrile 532.3 I-318 ##STR00374##
1-(4-(6-chloro-7-(2- fluoro-6-hydroxyphenyl)- 5-
(trifluoromethyl)quinazolin- 4-yl)piperazin-1- yl)prop-2-en-1-one
481.1
I-319 ##STR00375## 1-(4-(2-amino-6-chloro- 7-(5-methyl-1H-indazol-
4-yl)quinazolin-4- yl)piperazin-1-yl)prop-2- en-1-one 448.2 I-320
##STR00376## 1-(4-(6-chloro-8-fluoro-7- (5-methyl-1H-indazol-4-
yl)quinazolin-4-yl)-2- (hydroxymethyl)piperazin-
1-yl)prop-2-en-1-one 481.2 I-321 ##STR00377##
1-(4-(6-chloro-7-(2,4- difluorophenyl)-8- hydroxyquinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one 431.1 I-322 ##STR00378##
1-(4-(6-chloro-8-fluoro-7- (5-methyl-1H-indazol-4-
yl)-2-(1-methyl-1H- pyrazol-4- ylamino)quinaolin-4-
yl)piperazin-1-yl)prop-2- en-1-one 546.2 I-323 ##STR00379##
1-(4-(6-chloro-2-(2- (dimethylamino)ethoxy)-
8-fluoro-7-(5-methyl-1H- indazol-4-yl)quinazolin-
4-yl)piperazin-1-yl)prop- 2-en-1-one 538.2 I-324 ##STR00380##
1-(4-(6-chloro-8-fluoro-7- (3-methyl-1H-indazol-7- yl)quinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one 451.1 I-325 ##STR00381##
(E)-1-(4-(6-chloro-8- fluoro-7-(3- hydroxynaphthalen-1-
yl)quuinazolin-4- yl)piperazin-1-yl)-4- (dimethylamino)but-2-en-
1-one 520.3 I-326 ##STR00382## 4-(4-acryloylpiperazin-1-
yl)-6-chloro-7-(5-methyl- 1H-indazol-4- yl)quinazolin-2(1H)-one
448.2 I-327 ##STR00383## 1-(4-(6-chloro-8-fluoro-7-
(5-methyl-1H-indazol-4- yl)quinazolin-4-yl)-2- methylpiperazin-1-
yl)prop-2-en-1-one 465.2 I-328 ##STR00384## (E)-1-(4-(6-chloro-8-
fluoro-7-(3- hydroxynaphthalen-1- yl)quinazolin-4-yl)-2-
methylpiperazin-1-yl)but- 2-en-1-one 477.2 I-329 ##STR00385##
1-(4-(6-chloro-2-(3- (dimethylamino)propoxy)-
7-(5-methyl-1H-indazol- 4-yl)quinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one 534.3 I-330 ##STR00386##
1-(4-(6-chloro-7-(5- methyl-1H-indazol-4-yl)- 2-(tetrahydrofuran-3-
ylamino)quinazolin-4- yl)piperazin-1-yl)prop-2- en-1-one 518.3
I-331 ##STR00387## 1-(4-(6-chloro-8-fluoro-7-
(5-fluoro-1H-indazol-4- yl)quinazolin-4- yl)piperazin-1-yl)prop-2-
en-1-one 455.1 I-332 ##STR00388## 1-(4-(6-chloro-8-fluoro-7-
(5-methyl-1H-indazol-4- yl)-2-(thiazol-5- yl)quinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one 534.1 I-333 ##STR00389##
(E)-1-(4-(6-chloro-8- fluoro-7-(5-methyl-1H-
indazol-4-yl)-2-(thiazol- 5-yl)quinazolin-4- yl)piperazin-1-yl)-4-
(dimethylamino)but-2-en- 1-one 591.1 I-334 ##STR00390##
1-(4-(6-chloro-8-fluoro-7- (5-methyl-1H-indazol-4- yl)quinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one 451.2 I-335 ##STR00391##
(E)-4-amino-1-(4-(6- chloro-8-fluoro-7-(3- hydroxynaphthalen-1-
yl)quinazolin-4- yl)piperazin-1-yl)but-2- en-1-one 492.2 I-336
##STR00392## 4-(4-acryloyl-3- methylpiperazin-1-yl)-6-
chloro-8-fluoro-7-(5- methyl-1H-indazol-4- yl)quinoline-3-
carbonitrile 489.2 I-337 ##STR00393## 1-(4-(6-chloro-7-(3-
difluoromethyl)naphthalen- 1-yl)-8- fluoroquinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one 497.2 I-338 ##STR00394##
1-(4-(6-chloro-2- (dimethylamino)-8- fluoro-7-(5-methyl-1H-
indazol-4-yl)quinazolin- 4-yl)piperazin-1-yl)prop- 2-en-1-one 494.4
I-339 ##STR00395## 1-(4-(6-chloro-2-(3- (dimethylamino)propoxy)-
8-fluoro-7-(5-methyl- 1H-indazol-4- yl)quinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one 552.2 I-340 ##STR00396##
1-(4-(6-chloro-8-fluoro-7- (3-fluoro-5-methyl-1H-
indazol-4-yl)quinazolin- 4-yl)piperazin-1-yl)prop- 2-en-1-one 469.1
I-341 ##STR00397## 1-(4-(6-chloro-2-(2- (dimethylamino)ethoxy)-
8-fluoro-7-(2-fluoro-6- hydroxyphenyl)quinazolin- 4-yl)piperazin-1-
yl)prop-2-en-1-one 518.2 I-342 ##STR00398##
1-(4-(6-chloro-8-fluoro-7- (5-methyl-1H-indazol-4- yl)-2-(2-
morpholinoethoxy) quinazolin-4-yl)piperazin-1- yl)prop-2-en-1-one
580.2 I-343 ##STR00399## 1-(4-(6-chloro-5-fluoro-7- (2-fluoro-6-
hydroxyphenyl)quinazolin- 4-yl)piperazin-1- yl)prop-2-en-1-one
431.2 I-344 ##STR00400## 1-(4-(6-chloro-2- (dimethylamino)-8-
fluoro-7-(3- hydroxynaphthalen-1- yl)quinazolin-4-
yl)piperazin-1-yl)prop-2- en-1-one 506.3 I-345 ##STR00401##
1-(4-(6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1- yl)-2-
(methylamnio)quinazolin- 4-yl)piperazin-1-yl)prop- 2-en-1-one 492.2
I-346 ##STR00402## 1-(4-(6-chloro-8-fluoro-7-
(3-hydroxynaphthalen-1- yl)quinazolin-4-yl)-2-
((dimethylamino)methyl) piperazin-1-yl)prop-2-en- 1-one 520.2 I-347
##STR00403## 1-(4-(6-chloro-8-fluoro-7- (5-methyl-1H-indazol-4-
yl)-2-(tetrahydrofuran-3- ylamino)quinaolin-4-
yl)piperazin-1-yl)prop-2- en-1-one 536.3 I-348 ##STR00404##
1-(4-(6-chloro-7-(3- hydroxynaphthalen-1-yl)-
8-methoxyquinazolin-4- yl)piperazin-1-yl)prop-2- en-1-one 475.3
I-349 ##STR00405## 1-(4-(6-chloro-8-fluoro-7-
(3-hydroxynaphthalen-1- yl)-2-(1-(2,2,2- trifluoroethyl)pyrrolidin-
3-ylamino)quinazolin-4- yl)piperazin-1-yl)prop-2- en-1-one 629.3
I-350 ##STR00406## 1-(4-(7-(5-methyl-1H- indazol-4-yl)-6-
(trifluoromethyl)cinnolin- 4-yl)piperazin-1-yl)prop- 2-en-1-one
467.3 I-351 ##STR00407## 1-(4-(7-(3- hydroxynaphthalen-1-yl)- 6-
(trifluoromethyl)cinnolin- 4-yl)piperazin-1-yl)prop- 2-en-1-one
479.2 352 ##STR00408## 1-(4-(6-chloro-8- fluoro-2-(2-(1-methyl-
1H-imidazol-2- yl)ethylamino)-7-(5- methyl-1H-indazol-4-
yl)quinazolin-4- yl)piperazin-1-yl)prop- 2-en-1-one F 575.2 353
##STR00409## (S)-1-(4-(6-chloro-8- fluoro-7-(2-fluoro-6-
hydroxyphenyl)quinazolin- 4-yl)piperazin-1- yl)prop-2-en-1-one O
431.2 354 ##STR00410## 1-(4-(6-chloro-8- fluoro-7-(5-methyl-1H-
indazol-4-yl)-2-(1- (2,2,2- trifluoroethyl)pyrrolidin- 3-
ylamino)quinazolin-4- yl)piperazin-1-yl)prop- 2-en-1-one AC 617.3
355 ##STR00411## 1-(4-(6-chloro-2-(2- (dimethylamino)ethylamino)-
8-fluoro-7-(3- hydroxynaphthalen-1- yl)quinazolin-4-
yl)piperazin-1-yl)prop- 2-en-1-one AC 549.3 356 ##STR00412##
1-(4-(6-chloro-8- fluoro-7-(5-methyl-1H- indazol-4-yl)-2-(2,2,2-
trifluoroethylamino) quinazolin-4-yl)piperazin-
1-yl)prop-2-en-1-one AC 548.3 357 ##STR00413## 1-(4-(6-chloro-8-
fluoro-7-(5-methyl-1H- indazol-4-yl)-2-(3- morpholinopropoxy)
quinazolin-4-yl)piperazin- 1-yl)prop-2-en-1-one F 594.2 358
##STR00414## 1-(4-(6-chloro-8- fluoro-7-(5-methyl-1H-
indazol-4-yl)-2- (2- (pyrrolidin-1- yl)ethoxy)quinazolin-4-
yl)piperazin-1-yl)prop- 2-en-1-one F 564.2 359 ##STR00415##
1-(4-(6-chloro-8- fluoro-7-(6-fluoro-3- methyl-1H-indazol-7-
yl)quinazolin-4- yl)piperazin-1-yl)prop- 2-en-1-one B 469.1 360
##STR00416## 1-(4-(6-chloro-8- fluoro-7-(3- hydroxynaphthalen-1-
yl)-2-(1- methylpyrrolidin-3- ylamino)quinazolin-4-
yl)piperazin-1-yl)prop- 2-en-1-one AC 561.4 361 ##STR00417##
1-(4-(6-chloro-2-((2- (dimethylamino)ethyl) (methyl)amino)-8-
fluoro-7-(3- hydroxynaphthalen-1- yl)quinazolin-4-
yl)piperazin-1-yl)prop- 2-en-1-one AC 563.4 362 ##STR00418##
1-(4-(6-chloro-7-(2- ((dimethylamino)methyl)- 6-fluorophenyl)-8-
fluoroquinazolin-4- yl)piperazin-1-yl)prop- 2-en-1-one O 472.3 363
##STR00419## 1-(4-(6-chloro-8- fluoro-7-(5-methyl-1H-
indazol-4-yl)-2-(1- methylpiperidin-4- ylamino)quinazolin-4-
yl)piperazin-1-yl)prop- 2-en-1-one AC 563.4 364 ##STR00420##
1-(4-(6-chloro-8- fluoro-7-(3- hydroxynaphthalen-1- yl)-2-(3,3,3-
trifluoropropylamino) quinazolin-4- yl)piperazin-1-yl)prop-
2-en-1-one AC 560.30 365 ##STR00421## 1-(4-(6-chloro-8-
fluoro-7-(5-methyl-1H- indazol-4-yl)-2- (tetrahydro-2H-pyran-
4-ylamino)quinazolin- 4-yl)piperazin-1- yl)prop-2-en-1-one AC
550.30 366 ##STR00422## N-(2-(4-(4- acryloylpiperazin-1-yl)-
6-chloro-8-fluoro-7-(5- methyl-1H-indazol-4- yl)quinazolin-2-
yloxy)etthyl)acetamide AC 552.35 367 ##STR00423##
1-(4-(6-chloro-2-(2- (dimethylamino)ethoxy)- 8-fluoro-7-(5-methyl-
1H-indazol-4- yl)quinazolin-4-yl)-2- methylpiperazin-1-
yl)prop-2-en-1-one F 552.30 368 ##STR00424## 1-(4-(6-chloro-8-
fluoro-7-(6-fluoro-1H- indazol-7- yl)quinazolin-4-
yl)piperazin-1-yl)prop- 2-en-1-one B 455.1 369 ##STR00425##
1-(4-(6-chloro-8- fluoro-2-(2-(1-methyl- 1H-imidazol-2-
yl)ethoxy)-7-(5-methyl- 1H-indazol-4- yl)quinazolin-4-
yl)piperazin-1-yl)prop- 2-en-1-one F 575.2 370 ##STR00426##
(R)-1-(4-(6-chloro-8- fluoro-7-(2-fluoro-6-
hydroxyphenyl)quinazolin- 4-yl)piperazin-1- yl)prop-2-en-1-one O
431.2 371 ##STR00427## 1-(4-(6-chloro-2-((2- (dimethylamino)ethyl)
(methyl)amino)-8- fluoro-7-(5-methyl-1H- indazol-4-
yl)quinazolin-4- yl)piperazin-1-yl)prop- 2-en-1-one AC 551.35 372
##STR00428## 1-(4-(6-chloro-8- fluoro-7-(5-methyl-1H-
indazol-4-yl)-2-(1- methylpyrrolidin-3- ylamino)quinazolin-4-
yl)piperazin-1-yl)prop- 2-en-1-one AC 549.30 373 ##STR00429##
1-(4-(6-chloro-8- fluoro-7-(5-methyl-1H- indazol-4-yl)-2-(2-(4-
methylpiperazin-1- yl)ethoxy)quinazolin-4- yl)piperazin-1-yl)prop-
2-en-1-one F 593.30 374 ##STR00430## 1-(4-(6-chloro-2-(2-
(dimethylamino)etthoxy)- 8-fluoro-7-(3- hydroxynaphthalen-1-
yl)quinazolin-4- yl)piperazin-1-yl)prop- 2-en-1-one F 550.25 375
##STR00431## 1-(4-(6-chloro-8- fluoro-7-(5-methyl-1H-
indazol-4-yl)-2-(1- methyl-1H-pyrazol-4- yloxy)quinazolin-4-
yl)piperazin-1-yl)prop- 2-en-1-one F 547.25 376 ##STR00432##
4-(4-acryloylpiperazin- 1-yl)-6-chloro-8-fluoro- 7-(3-
hydroxynaphthalen-1- yl)quinazoline-2- carbonitrile AI 488.15 *[M +
Na].sup.+ .sup.+[M - H].sup.- .sup.#[M] Prepared according to
methods analogous to those illustrated herein ##STR00433##
[0393] The compounds in Table 1 were each prepared and analyzed by
mass spectrometry and/or .sup.1H NMR. Experimental mass
spectrometry data is included in Table 1 above. Exemplary synthetic
procedures are described in more detail below and in the Examples.
General methods by which the compounds may be prepared are provided
below and indicated in Table 1 above.
[0394] It is understood that in the present description,
combinations of substituents and/or variables of the depicted
formulae are permissible only if such contributions result in
stable compounds.
[0395] It will also be appreciated by those skilled in the art that
in the processes for preparing the compounds described herein the
functional groups of intermediate compounds may need to be
protected by suitable protecting groups. Such functional groups
include, but are not limited to, hydroxy, amino, mercapto and
carboxylic acid.
[0396] Suitable protecting groups for hydroxy include trialkylsilyl
or diarylalkylsilyl (for example, t-butyldimethylsilyl,
t-butyldiphenylsilyl or trimethylsilyl), tetrahydropyranyl, benzyl,
and the like. Suitable protecting groups for amino, amidino and
guanidino include t-butoxycarbonyl, benzyloxycarbonyl, and the
like. Suitable protecting groups for mercapto include --C(O)--R''
(where R'' is alkyl, aryl or arylalkyl), p-methoxybenzyl, trityl
and the like. Suitable protecting groups for carboxylic acid
include alkyl, aryl or arylalkyl esters.
[0397] Protecting groups are optionally added or removed in
accordance with standard techniques, which are known to one skilled
in the art and as described herein. The use of protecting groups is
described in detail in Green, T. W. and P. G. M. Wutz, Protective
Groups in Organic Synthesis (1999), 3rd Ed., Wiley. As one of skill
in the art would appreciate, the protecting group may also be a
polymer resin such as a Wang resin, Rink resin or a
2-chlorotrityl-chloride resin.
[0398] It will also be appreciated by those skilled in the art,
although such protected derivatives of compounds of this invention
may not possess pharmacological activity as such, they may be
administered to a subject and thereafter metabolized in the body to
form compounds of the invention which are pharmacologically active.
Such derivatives may therefore be described as "prodrugs". All
prodrugs of compounds of this invention are included within the
scope of the invention.
[0399] Furthermore, all compounds of the invention which exist in
free base or acid form can be converted to their pharmaceutically
acceptable salts by treatment with the appropriate inorganic or
organic base or acid by methods known to one skilled in the
art.
[0400] Salts of the compounds of the invention can be converted to
their free base or acid form by standard techniques.
[0401] The following General Reaction Schemes illustrate exemplary
methods of making compounds of compounds of structure (I):
##STR00434##
or a pharmaceutically acceptable salt, tautomer or stereoisomer
thereof, wherein R.sup.1, R.sup.2a, R.sup.3a, R.sup.3b, R.sup.4a,
R.sup.4b, G.sup.1, G.sup.2, L.sup.1, L.sup.2, m.sup.1, m.sup.2, A,
B, W, X, Y, Z and E are as defined above. For ease of illustration,
many of the schemes which follow illustrate an "R.sup.2" moiety.
The R2 moiety is meant to include any one of R.sup.2a, R.sup.2b or
R.sup.2c. It is understood that one skilled in the art may be able
to make these compounds by similar methods or by combining other
methods known to one skilled in the art. It is also understood that
one skilled in the art would be able to make, in a similar manner
as described below, other compounds of structure (I) not
specifically illustrated below by using the appropriate starting
components and modifying the parameters of the synthesis as needed.
In general, starting components may be obtained from sources such
as Sigma Aldrich, Lancaster Synthesis, Inc., Maybridge, Matrix
Scientific, TCI, and Fluorochem USA, etc. or synthesized according
to sources known to those skilled in the art (see, for example,
Advanced Organic Chemistry: Reactions, Mechanisms, and Structure,
5th edition (Wiley, December 2000)) or prepared as described in
this invention.
##STR00435## ##STR00436##
[0402] Embodiments of the compound of structure (I) (e.g., compound
A-7) can be prepared according to General Reaction Scheme 1
("Method A"), wherein R.sup.1, R.sup.2, R.sup.3a, R.sup.3bR.sup.4a,
R.sup.4b, R.sup.9, R.sup.10, Q, m.sup.1 and m.sup.2 are as defined
herein above. As shown in General Reaction Scheme 1, compounds of
structure A-1 can be purchased from commercial sources or prepared
according to methods familiar to one of ordinary skill in the art.
Reaction of A-1 under Suzuki conditions yields A-2. Reaction of
compounds of structure A-2 with formamide or other suitable
reagents, such as formamidine acetate or trimethyl orthoformate,
yields quinazolines of structure A-3. A-3 is chlorinated under
appropriate conditions (e.g., SOCl.sub.2, POCl.sub.3/PCl.sub.5 or
POCl.sub.3) to yield chloroquinazoline A-4. Reaction of A-4 with an
appropriately protected heterocycle under basic conditions yields
A-5. Appropriate protecting groups include butyloxycarbonyl (BOC)
as depicted in General reaction Scheme 1, as well as other
protecting groups known in the art. Deprotection of A-5 followed by
acylation with an acid chloride (or sulfonyl chloride) or acid and
appropriate activating reagents yields A-7.
##STR00437##
[0403] Alternatively, embodiments of the compound of structure (I)
(e.g., compound A-7) can be prepared according to General Reaction
Scheme 2 ("Method B"), wherein R.sup.1, R.sup.2, R.sup.3a,
R.sup.3b, R.sup.4a, R.sup.4b, R.sup.9, R.sup.10, Q, m.sup.1 and
m.sup.2 are as defined herein above. Compounds of structure A-1 are
prepared or purchased as described above. Treatment of A-1 with
formamide or other suitable reagents, such as formamidine acetate
or trimethyl orthoformate, yields quinazolines of structure B-1.
B-1 can then be chlorinated to yield B-2 and reacted with an
appropriately protected heterocycle under basic conditions to yield
B-3 as described above for Method A. Suzuki coupling then yields
A-5 which can be converted to A-7 as described in Method A
above.
##STR00438## ##STR00439##
[0404] Other embodiments of the compound of structure (I) (e.g.,
compound C-6) can be prepared according to General Reaction Scheme
3 ("Method C"), wherein R.sup.1, R.sup.2, R.sup.3a, R.sup.3b,
R.sup.4a, R.sup.4b, R.sup.9, R.sup.10, Q, m.sup.1 and m.sup.2 are
as defined herein above. As shown in General Reaction Scheme 3,
compounds of structure C-1, which can be purchased from commercial
sources or prepared according to well-known procedures, are reacted
with tosyl hydrazine to yield C-2. Chlorination of C-2 with an
appropriate reagent(s), such as thionyl chloride, then yields C-3
which can be reacted under basic conditions with an appropriately
protected heterocycle (PG=protecting group or C.sub.1-C.sub.6alkyl)
to yield indazole C-4. The tosyl group is removed from C-4 by
treatment with sodium hydroxide in THF/H.sub.2O to yield C-5.
Removal of the nitrogen protecting group and acylation or
thioacylation as described in Method A then yields the desired
compound C-6.
##STR00440## ##STR00441##
[0405] Other embodiments of the compound of structure (I) (e.g.,
compound D-9) can be prepared according to General Reaction Scheme
4 ("Method D"), wherein R.sup.1, R.sup.2, R.sup.3a, R.sup.3b,
R.sup.4a, R.sup.4b, R.sup.9, R.sup.10, Q, m.sup.1 and m.sup.2 are
as defined herein above. As shown in General Reaction Scheme 4,
benzaldehyde D-1 is treated under reductive amination conditions to
yield D-2. Formation of the tosyl-protected amine (D-3) followed by
treatment with an appropriate Lewis acid (e.g., AlCl.sub.3) yields
isoquinoline D-4. Oxidation of D-4 with meta-chloroperbenzoic acid
(mCPBA) yields D-5 which can be chlorinated by treatment with an
appropriate reagent, such as POCl.sub.3. Chloride D-6 is then
treated in a manner analogous to that described for Method B to
yield D-9.
##STR00442## ##STR00443##
[0406] Other embodiments of the compound of structure (I) (e.g.,
compound E-9) can be prepared according to General Reaction Scheme
5 ("Method E"), wherein R.sup.1, R.sup.2, R.sup.3a, R.sup.3b,
R.sup.4a, R.sup.4b, R.sup.9, R.sup.10, Q, m.sup.1 and m.sup.2 are
as defined herein above. As shown in General Reaction Scheme 5,
aniline E-1, which can be purchased from commercial sources or
prepared via well-known procedures, can be reacted with diethyl
2-(ethoxymethylene)malonate to yield E-2. E-2 can then be cyclized
by heating in an appropriate high-boiling solvent (e.g., Ph.sub.2O)
to yield quinolone E-3. Saponification of E-3 followed by
decarboxylation yields E-4 and E-5, respectively. E-5 is then
treated in a manner analogous to that described for Method B to
yield E-9.
##STR00444## ##STR00445##
[0407] Other embodiments of the compound of structure (I) (e.g.,
compound F-6) can be prepared according to General Reaction Scheme
6 ("Method F"), wherein R.sup.1, R.sup.2, R.sup.3a, R.sup.3b,
R.sup.4a, R.sup.4b, R.sup.9, R.sup.10, Q, m.sup.1 and m.sup.2 are
as defined herein above. As shown in General Reaction Scheme 6, A-1
is cyclized to quinazolinedione F-1 by treatment with urea.
Chlorination of F-1 by treatment with POCl.sub.3 followed by
reaction with a protected heterocycle yield F-2 and F-3,
respectively. The R.sup.6 substituent is installed by S.sub.NAr
reaction of G-3 with LG-R.sup.6, wherein LG is an appropriate
leaving group. For example, where R.sup.6 is cyano or alkoxy, LG is
sodium or another appropriate action. The general procedures
described above with respect to Method B can then be employed to
yield F-6.
##STR00446##
[0408] Other embodiments of the compound of structure (I) (e.g.,
compound G-4) can be prepared according to General Reaction Scheme
7 ("Method G"), wherein R.sup.1, R.sup.2, R.sup.3a, R.sup.3b,
R.sup.4a, R.sup.4b, R.sup.9, R.sup.10, Q, m.sup.1 and m.sup.2 are
as defined herein above. As shown in General Reaction Scheme 7,
aniline E-1 is treated under Suzuki conditions to install the R-1
substituent. G-1 is then heated in toluene with an appropriately
substituted unsaturated ester to yield G-2. Cyclization of G-2 to
hydroxyquinoline G-3 is accomplished by heating in a high boiling
solvent (e.g., Ph.sub.2O) for an appropriate amount of time.
Following the general procedures outlined in Method A then yields
G-4.
##STR00447##
[0409] Other embodiments of the compound of structure (I) (e.g.,
compound H-3) can be prepared according to General Reaction Scheme
8 ("Method H"), wherein R.sup.1, R.sup.2, R.sup.3a, R.sup.3b,
R.sup.4a, R.sup.4b, R.sup.9, R.sup.10, Q, m.sup.1 and m.sup.2 are
as defined herein above. Referring to General Reaction Scheme 8,
thienopyrimidine H-1 can be prepared according to well-known
procedures or purchased from commercial sources. H-1 is treated
with an appropriately protected heterocycle under basic conditions
to yield H-2. Deprotection followed by acylation or thioacylation
according to the procedures described above then yields H-3.
##STR00448##
[0410] Other embodiments of the compound of structure (I) (e.g.,
compound I-4) can be prepared according to General Reaction Scheme
9 ("Method I"), wherein R.sup.1, R.sup.2, R.sup.3a, R.sup.3b,
R.sup.4a, R.sup.4b, R.sup.9, R.sup.10, Q, m.sup.1 and m.sup.2 are
as defined herein above. Referring to General Reaction Scheme 9,
quinazoline I-1 can be prepared according to well-known procedures
or purchased from commercial sources. I-1 is treated with an
appropriately protected heterocycle under basic conditions to yield
I-2. Suzuki reaction of I-2 with an appropriate reagent to install
the R.sup.1 moiety results in I-3. I-3 is then deprotected and
acylated (or thioacylated) according to the procedures described
above to yield I-4.
##STR00449##
[0411] Other embodiments of the compound of structure (I) (e.g.,
compound J-6) can be prepared according to General Reaction Scheme
10 ("Method J"), wherein R.sup.1, R.sup.2, R.sup.3a, R.sup.3b,
R.sup.4a, R.sup.4b, R.sup.9, R.sup.10, Q, m.sup.1 and m.sup.2 are
as defined herein above. Referring to General Reaction Scheme 10,
pyrrolopyrimidinone J-1 can be prepared according to well-known
procedures or purchased from commercial sources. J-1 is chlorinated
with an appropriate reagent (e.g., POCl3) to yield J-2 which is
then iodinated with an appropriate reagent, such as
N-iodosuccinimide (NIS) to yield J-3. Protection of J-3 followed by
Suzuki reaction yields J-5. J-5 is then treated according to the
procedures described above to yield J-6.
##STR00450##
[0412] Other embodiments of the compound of structure (I) (e.g.,
compound K-5) can be prepared according to General Reaction Scheme
11 ("Method K"), wherein R.sup.1, R.sup.2, R.sup.3a, R.sup.3b,
R.sup.4a, R.sup.4b, R.sup.9, R.sup.10, Q, m.sup.1 and m.sup.2 are
as defined herein above. Referring to General Reaction Scheme 11,
quinazoline K-1 can be prepared according to well-known procedures
or purchased from commercial sources. K-1 is reacted with an
appropriate ester under basic conditions to form the requisite
carbon-carbon bond. K-2 is then decarboxylated to yield K-3. Suzuki
reaction, deprotection and acylation or thioacylation are then
carried out as described in the above schemes to yield K-5.
##STR00451##
[0413] Other embodiments of the compound of structure (I) (e.g.,
compound L-2) can be prepared according to General Reaction Scheme
12 ("Method L"), wherein R.sup.1, R.sup.2, R.sup.3a, R.sup.3b,
R.sup.4a, R.sup.4b, R.sup.9, R.sup.10, Q, m.sup.1 and m.sup.2 are
as defined herein above. Specifically, compounds wherein R.sup.1 is
an N-heterocycle can be efficiently prepared according to Method L.
Referring to General Reaction Scheme 12, compound B-3 is prepared
according to Method B and treated under Buchwald conditions (where
R.sup.1--H is a N-heterocycle or alkylamino) to yield L-1. Methods
for Buchwald reactions are well-known in the art. L-1 is then
converted to L-2 according to the above general procedures.
##STR00452##
[0414] Other embodiments of the compound of structure (I) (e.g.,
compound M-3) can be prepared according to General Reaction Scheme
13 ("Method M"), wherein R.sup.1, R.sup.2, R.sup.3a, R.sup.3b,
R.sup.4a, R.sup.4b, R.sup.6, R.sup.9, R.sup.10, Q, m.sup.1 and
m.sup.2 are as defined herein above. Referring to General Reaction
Scheme 13, compound A-1 is reacted with an appropriate nitrile
(R.sup.6CN) to form compound M-1. In this regard, R.sup.6 may be
any of the R.sup.6 moieties described herein, for example alkyl.
M-1 is chlorinated by reaction with an appropriate reagent such as
thionyl chloride. Compound M-3 is then prepared according to the
general procedures outlined herein, for example the procedures of
General Reaction Scheme 2.
##STR00453##
[0415] Embodiments of the compound of structure (I) (e.g., compound
N-7) can be prepared according to General Reaction Scheme 14
("Method N"), wherein R.sup.1, R.sup.2, R.sup.3a, R.sup.3b,
R.sup.4a, R.sup.4b, R.sup.9, R.sup.10, Q, m.sup.1 and m.sup.2 are
as defined herein above. As shown in General Reaction Scheme 14,
compounds of structure N-1 can be purchased from commercial sources
or prepared according to methods familiar to one of ordinary skill
in the art. Compound N-1 is reacted with methylnitrile to form
compound N-2. Reaction of N-2 with sodium nitrite under acidic
conditions yields cinnolines of structure N-3. N-3 is chlorinated
under appropriate conditions (e.g., SOCl.sub.2,
POCl.sub.3/PCl.sub.5 or POCl.sub.3) to yield the chlorocinnoline
N-4. Reaction of N-4 with an appropriately protected heterocycle
under basic conditions yields N-5. Appropriate protecting groups
include butyloxycarbonyl (BOC) as depicted in General reaction
Scheme 1, as well as other protecting groups known in the art.
Suzuki reaction of N-5 with an appropriate reagent to install the
R.sup.1 moiety results in N-6. Deprotection of N-6 followed by
acylation with an acid chloride (or sulfonyl chloride) or acid and
appropriate activating reagents yields N-7.
##STR00454## ##STR00455##
[0416] Embodiments of the compound of structure (I) (e.g., compound
O-11) can be prepared according to General Reaction Scheme 15
("Method O"), wherein R.sup.1, R.sup.2b, R.sup.3a, R.sup.3b,
R.sup.4a, R.sup.4b, R.sup.9, R.sup.10, Q, m.sup.1 and m.sup.2 are
as defined herein above. As shown in General Reaction Scheme 15,
compounds of structure O-1 can be purchased from commercial sources
or prepared according to methods familiar to one of ordinary skill
in the art. Compound O-1 is reduced to form compound O-2. Reaction
of O-2 with 2,2,2-trichloroethane-1,1-diol under acidic conditions,
then hydroxylamine hydrochloride, yields O-3. O-3 is cyclized in
the presence of acid to yield O-4. O-4 is reacted in the presence
H.sub.2O.sub.2 under basic conditions to yield O-5. O-5 is
chlorinated using N-chlorosuccinimide to yield O-6. Reaction of O-6
with formamide or other suitable reagents such as formamidine
acetate or trimethyl orthoformate yields the quinazolin-4(3H)-one,
O-7. O-7 is chlorinated under appropriate conditions (e.g.,
SOCl.sub.2, POCl.sub.3/PCl.sub.5 or POCl.sub.3) to yield the
chloroquinazoline, O-8. Reaction of O-8 with an appropriately
protected heterocycle under basic conditions yields O-9.
Appropriate protecting groups include butyloxycarbonyl (BOC) as
depicted in General reaction Scheme 1, as well as other protecting
groups known in the art. Suzuki reaction of O-9 with an appropriate
reagent to install the R.sup.1 moiety results in O-10. Deprotection
of O-10 followed by acylation with an acid chloride (or sulfonyl
chloride) or acid and appropriate activating reagents yields
O-11.
##STR00456## ##STR00457## ##STR00458##
[0417] Embodiments of the compound of structure (I) (e.g., compound
P-10) can be prepared according to General Reaction Scheme 16
("Method P"), wherein R.sup.1, R.sup.2b, R.sup.3aR.sup.3b,
R.sup.4a, R.sup.4b, R.sup.9, R.sup.10, Q, m.sup.1 and m.sup.2 are
as defined herein above. As shown in General Reaction Scheme 16,
compound 0-2 is chlorinated using N-chlorosuccinimide to yield P-1.
Reaction of P-1 with diethyl-2-(ethoxymethylene)malonate yields
P-2. P-2 is then cyclized by heating in an appropriate high-boiling
solvent (e.g. Ph.sub.2O) to yield the quinolone, P-3. P-3 is
chlorinated under appropriate conditions (e.g., SOCl.sub.2,
POCl.sub.3/PCl.sub.5 or POCl.sub.3) to yield the chloroquinolone,
P-4. Reaction of P-4 with an appropriately protected heterocycle
under basic conditions yields P-5. Appropriate protecting groups
include butyloxycarbonyl (BOC) as depicted in General reaction
Scheme 1, as well as other protecting groups known in the art.
Saponification of P-5 followed by amidation yields P-6 and P-7,
respectively. Suzuki reaction of P-7 with an appropriate reagent to
install the R.sup.1 moiety results in P-8. Deprotection of P-8
followed by acylation with an acid chloride (or sulfonyl chloride)
or acid and appropriate activating reagents yields P-9. Reaction of
P-9 in the presence of acid yielded P-10.
##STR00459##
[0418] Embodiments of the compound of structure (I) (e.g., compound
Q-2) can be prepared according to General Reaction Scheme 16
("Method Q"), wherein R.sup.1, R.sup.2b, R.sup.3aR.sup.3b,
R.sup.4a, R.sup.4b, R.sup.9, R.sup.10, Q, m.sup.1 and m.sup.2 are
as defined herein above. As shown in General Reaction Scheme 17,
deprotection of compound 0-9 followed by acylation with an acid
chloride (or sulfonyl chloride) or acid and appropriate activating
reagents yields Q-1. Suzuki reaction of Q-1 with an appropriate
reagent to install the R.sup.1 moiety results in Q-2. One skilled
in the art will recognize that certain modifications to the above
schemes are possible to prepare different embodiments of compounds
of structure (I). For example, for ease of illustration, most of
the above schemes depict preparation of compounds of structure (I)
wherein L.sup.1 is a bond. However, one of ordinary skill in the
art will readily recognize that compounds wherein L.sup.1 is
NR.sup.7 can be prepared by substituting a heterocycle having the
following structure (see e.g., Method C):
##STR00460##
where R is H, a protecting group or C.sub.1-C.sub.6 alkyl.
2. Compounds of Structure (II)
[0419] In still other embodiments, the compound used in combination
with one or more additional therapeutic agent has the following
structure (II):
##STR00461##
or a pharmaceutically acceptable salt, tautomer, stereoisomer or
prodrug thereof, wherein:
[0420] R.sup.1 is aryl or heteroaryl;
[0421] R.sup.30a and R.sup.30b are, at each occurrence,
independently H, --OH, --NH.sub.2, --CO.sub.2H, cyano, cyanoalkyl,
C.sub.1-C.sub.6alkyl, C.sub.3-C.sub.8cycloalkyl, hydroxylalkyl,
aminoalkyl, carboxylalkyl or aminocarbonyl; or R.sup.30a and
R.sup.30b join to form a carbocyclic or heterocyclic ring; or
R.sup.30a is H, --OH, --NH.sub.2, --CO.sub.2H, cyano,
C.sub.1-C.sub.6alkyl, C.sub.3-C.sub.8cycloalkyl, hydroxylalkyl,
aminoalkyl, carboxylalkyl or aminocarbonyl and R.sup.30b joins with
R.sup.31b to form a carbocyclic or heterocyclic ring;
[0422] R.sup.31a and R.sup.31b are, at each occurrence,
independently H, --OH, --NH.sub.2, --CO.sub.2H, cyano, cyanoalkyl,
C.sub.1-C.sub.6alkyl, C.sub.3-C.sub.8cycloalkyl, hydroxylalkyl,
aminoalkyl, carboxylalkyl or aminocarbonyl; or R.sup.31a and
R.sup.31b join to form a carbocyclic or heterocyclic ring; or
R.sup.31a is H, --OH, --NH.sub.2, --CO.sub.2H, cyano,
C.sub.1-C.sub.6alkyl, C.sub.3-C.sub.8cycloalkyl, hydroxylalkyl,
aminoalkyl, carboxylalkyl or aminocarbonyl and R.sup.31b joins with
R.sup.30b to form a carbocyclic or heterocyclic ring;
[0423] R.sup.32a and R.sup.32b are, at each occurrence,
independently H, --OH, --NH.sub.2, --CO.sub.2H, cyano, cyanoalkyl,
C.sub.1-C.sub.6alkyl, C.sub.3-C.sub.8cycloalkyl, hydroxylalkyl,
aminoalkyl, carboxylalkyl or aminocarbonyl; or R.sup.32a and
R.sup.32b join to form a carbocyclic or heterocyclic ring; or
R.sup.32a is H, --OH, --NH.sub.2, --CO.sub.2H, cyano,
C.sub.1-C.sub.6alkyl, C.sub.3-C.sub.8cycloalkyl, hydroxylalkyl,
aminoalkyl, carboxylalkyl or aminocarbonyl and R.sup.32b joins with
R.sup.33b to form a carbocyclic or heterocyclic ring;
[0424] R.sup.33a and R.sup.33b are, at each occurrence,
independently H, --OH, --NH.sub.2, --CO.sub.2H, cyano, cyanoalkyl,
C.sub.1-C.sub.6alkyl, C.sub.3--C cycloalkyl, hydroxylalkyl,
aminoalkyl, carboxylalkyl or aminocarbonyl; or R.sup.33a and
R.sup.33b join to form a carbocyclic or heterocyclic ring; or
R.sup.33a is H, --OH, --NH.sub.2, --CO.sub.2H, cyano,
C.sub.1-C.sub.6alkyl, C.sub.3-C.sub.8cycloalkyl, hydroxylalkyl,
aminoalkyl, carboxylalkyl or aminocarbonyl and R.sup.33b joins with
R.sup.32b to form a carbocyclic or heterocyclic ring;
[0425] L.sup.1 is carbonyl, --NHC(.dbd.O)--, alkylene, alkenylene,
heteroalkylene, heterocycloalkylene, heteroarylene,
alkylenecarbonyl, alkenylenecarbonyl, heteroalkylenecarbonyl,
heterocycloalkylenecarbonyl or heteroarylenecarbonyl;
[0426] L.sup.2 is a bond or alkylene;
[0427] G.sup.1, G.sup.2, G.sup.3 and G.sup.4 are each independently
N or CR, where R is H, cyano, halo or C.sub.1-C.sub.6alkyl;
[0428] n.sup.1, n.sup.2, n.sup.3 and n.sup.4 are each independently
1, 2 or 3; and
[0429] E is an electrophilic moiety capable of forming a covalent
bond with the cysteine residue at position 12 of a KRAS, HRAS or
NRAS G12C mutant protein.
[0430] In some embodiments of the compounds of structure II,
L.sup.1 is carbonyl, --NHC(.dbd.O)--, alkylene, heteroalkylene,
alkylenecarbonyl or heteroalkylenecarbonyl;
[0431] In some other embodiments, the compound has the following
structure (IIa):
##STR00462##
wherein:
[0432] L.sup.1a is a bond, --NH--, alkylene, alkenylene,
heteroalkylene, heterocycloalkylene or heteroarylene.
[0433] In other embodiments of compound (IIa), L.sup.1a is a bond,
--NH--, alkylene or heteroalkylene.
[0434] In some more embodiments, the compound has the following
structure (IIb):
##STR00463##
wherein:
[0435] Q is --C(.dbd.O)--, --NR.sup.34C(.dbd.O)--,
--S(.dbd.O).sub.2-- or --NR.sup.34S(.dbd.O).sub.2--;
[0436] R.sup.34 is H, C.sub.1-C.sub.6alkyl or hydroxylalkyl;
[0437] is a carbon-carbon double bond or a carbon-carbon triple
bond; and
[0438] R.sup.35 and R.sup.36 are each independently H, cyano,
C.sub.1-C.sub.6alkyl, aminoalkyl, alkylaminoalkyl, or hydroxylalkyl
or R.sup.35 and R.sup.36 join to form a carbocyclic or heterocyclic
ring when is a double bond; or R.sup.35 is absent and R.sup.36 is
H, C.sub.1-C.sub.6alkyl, aminoalkyl, alkylaminoalkyl or
hydroxylalkyl when is a triple bond.
[0439] In some different embodiments, the compound has one of the
following structures (IIc), (IId), (IIe) or (IIf):
##STR00464## ##STR00465##
[0440] In still other embodiments, wherein the compound has one of
the following structures (IIg), (IIh), (IIi) or (IIj):
##STR00466##
[0441] In some other embodiments, the compound has one of the
following structures (Ilk), (IIl), (IIm), (IIn); (IIo) or
(IIp):
##STR00467## ##STR00468##
[0442] In various other embodiments, R.sup.1 is aryl. For example,
in some embodiments the aryl is bicyclic, such as a fused bicyclic
aryl. In some more specific embodiments, the aryl is naphthyl.
[0443] In various other embodiments, the aryl is monocyclic. For
example, in some embodiments the aryl is phenyl.
[0444] In some of the foregoing embodiments, the aryl is
unsubstituted. In other of the foregoing embodiments, the aryl is
substituted with one or more substituents. For example, in some
embodiments the substituents are selected from halo, hydroxyl,
cyano, aminocarbonyl, formyl, C.sub.1-C.sub.6alkyl,
C.sub.1-C.sub.6alkylsulfonyl, C.sub.1-C.sub.6haloalkyl,
C.sub.3-C.sub.8cycloalkyl, C.sub.1-C.sub.6alkoxy,
C.sub.1-C.sub.6hydroxylalkyl, C.sub.1-C.sub.6alkoxyalkyl,
C.sub.1-C.sub.6aminoalkyl, aliphatic heterocyclyl, heteroaryl and
aryl.
[0445] In other embodiments, the aryl substituents are selected
from fluoro, chloro, bromo, iodo, hydroxyl, cyano, methyl, ethyl,
isopropyl, methylsulfonyl, methoxy, aminocarbonyl, trifluoromethyl,
2,2,2-trifluorethyl, cyclobutyl, cyclopropyl and phenyl, wherein
the cyclopropyl and phenyl are optionally substituted with one or
more substituents selected from C.sub.1-C.sub.6alkyl, halo,
hydroxyl and cyano
[0446] In some different embodiments, the substituents are selected
from fluoro, chloro, bromo, iodo, hydroxyl, cyano, methyl, ethyl,
methylsulfonyl, methoxy, aminocarbonyl, trifluoromethyl,
cyclopropyl and phenyl, wherein the cyclopropyl and phenyl are
optionally substituted with one or more substituents selected from
halo, hydroxyl and cyano.
[0447] In other exemplary embodiments, the aryl substituents are
selected from fluoro, chloro, bromo, iodo, hydroxyl, methyl, ethyl,
cyclobutyl and cyclopropyl, wherein the cyclopropyl is optionally
substituted with one or more substituents selected from
C.sub.1-C.sub.6alkyl, halo, hydroxyl and cyano
[0448] In some more embodiments, the substituents are selected from
fluoro, chloro, bromo, iodo, hydroxyl, methyl, ethyl and
cyclopropyl, wherein the cyclopropyl is optionally substituted with
one or more substituents selected from halo, hydroxyl and
cyano.
[0449] In still more embodiments, the substituents are selected
from fluoro, chloro, bromo, hydroxyl and cyclopropyl, wherein the
cyclopropyl is optionally substituted with one or more substituents
selected from C.sub.1-C.sub.6alkyl, halo, hydroxyl and cyano.
[0450] In some more specific embodiments, the substituents are
selected from fluoro, chloro, bromo, hydroxyl and cyclopropyl,
wherein the cyclopropyl is optionally substituted with one or more
substituents selected from halo, hydroxyl and cyano. For example,
in some embodiments the cyclopropyl comprises a geminal difluoro
substitution.
[0451] In still other embodiments, R.sup.1 has one of the following
structures:
##STR00469## ##STR00470## ##STR00471##
[0452] In still other embodiments, R.sup.1 has one of the following
structures:
##STR00472## ##STR00473## ##STR00474##
[0453] In still other embodiments, R.sup.1 is heteroaryl. For
example, in some embodiments the heteroaryl is bicyclic, such as a
fused bicyclic heteroaryl.
[0454] In some more embodiments, the heteroaryl is monocyclic.
[0455] In some of the foregoing embodiments, the heteroaryl
comprises nitrogen, sulfur or a combination thereof. For example,
in some embodiments the heteroaryl is dihydroquinoxalinyl,
indoleyl, benzoimidazolyl, pyridinyl or thiazolyl.
[0456] In some embodiments, the heteroaryl is unsubstituted. In
some other embodiments, the heteroaryl is substituted with one or
more substituents. In some embodiments, the substituents are
selected from C.sub.1-C.sub.6alkyl, halo and oxo. For example, in
some embodiments the substituents are selected from halo and oxo.
In other embodiments, the substituents are selected from ethyl and
chloro. In some more specific embodiments, the substituents are
chloro.
[0457] In some embodiments of the forgoing compounds of structure
(II), R.sup.1 has one of the following structures:
##STR00475##
wherein R.sup.1a is, at each occurrence, independently H,
C.sub.1-C.sub.6alkyl or halo.
[0458] In various other embodiments, R.sup.1 has one of the
following structures:
##STR00476##
wherein R.sup.1a is, at each occurrence, independently H or
halo.
[0459] In still other embodiments of structure (II), R.sup.1 has
one of the following structures:
##STR00477##
[0460] In some embodiments, Q is --C(.dbd.O)--. In some other
embodiments, Q is --S(.dbd.O).sub.2--. In still other embodiments,
Q is --NR.sup.34C(.dbd.O)--. In still more other embodiments, Q is
--NR.sup.34S(.dbd.O).sub.2--.
[0461] In some more specific embodiments, R.sup.34 is H. For
example, in some embodiments R.sup.34 is hydroxylalkyl, such as
2-hydroxylalkyl.
[0462] In other of the foregoing embodiments, at least one of
R.sup.35 or R.sup.36 is H. For example, in some embodiment search
of R.sup.35 and R.sup.36 are H.
[0463] In various other embodiments, R.sup.36 is alkylaminoalkyl.
For example, in some embodiments R.sup.36 has the following
structure:
##STR00478##
[0464] In some different embodiments, R.sup.36 is hydroxylalkyl,
for example 2-hydroxylalkyl
[0465] In various other embodiments, R.sup.35 and R.sup.36 join to
form a ring. In some of these embodiments, the ring is a
cyclopentene, cyclohexene or phenyl ring.
[0466] In other of the foregoing embodiments, E has one of the
following structures:
##STR00479##
[0467] In some embodiments, E is
##STR00480##
[0468] In some more of the foregoing embodiments, L.sup.1 is
heteroalkylene. In some more embodiments, the heteroalkylene is
unsubstituted. In some different embodiments, the heteroalkylene is
substituted.
[0469] In various other embodiments, L is aminoalkylene. For
example, in some embodiments L.sup.1 is --CH.sub.2CH.sub.2NH--.
[0470] In other embodiments of the foregoing, L.sup.1 is
heterocycloalkylene or heteroarylene. In some embodiments, the
heterocycloalkylene or heteroarylene is unsubstituted. In other
embodiments, the heterocycloalkylene or heteroarylene is
substituted. In some further embodiments, L.sup.1 has one of the
following structures:
##STR00481##
[0471] In some different embodiments, L.sup.1a is a bond.
[0472] In some embodiments, L.sup.1a is alkylene, alkenylene,
heteroalkylene or heterocycloalkylene. In some other embodiments,
L.sup.1a is alkylene or heteroalkylene. In some of these
embodiments, L.sup.1a is substituted alkylene. In various other
embodiments, L.sup.1a is unsubstituted alkylene. For example, in
some embodiments L.sup.1a is
##STR00482##
[0473] In some different embodiments, L.sup.1a is substituted
heteroalkylene. In some other embodiments, L.sup.1a is
unsubstituted heteroalkylene. In some of the foregoing embodiments,
L.sup.1a is aminoalkylene or thioalkylene, for example
aminoalkylene. For example, in some embodiments L.sup.1a has one of
the following structures:
##STR00483##
[0474] In other embodiments, L.sup.1a is
##STR00484##
[0475] In other embodiments, L.sup.1a is substituted alkenylene. In
different embodiments, L.sup.1a is unsubstituted alkenylene. In
some more specific embodiments, L.sup.1a has the following
structure:
##STR00485##
[0476] In yet other embodiments, L.sup.1a is substituted
heterocycloalkylene. In some other embodiments, L.sup.1a is
unsubstituted heterocycloalkylene. For Example, in some
embodiments, L.sup.1a has the following structure:
##STR00486##
[0477] In some of the foregoing embodiments, L.sup.2 is a bond.
[0478] In various other embodiments, L.sup.2 is substituted
alkylene. In still other embodiments, L.sup.2 is unsubstituted
alkylene.
[0479] In various embodiments of any of the foregoing compounds of
structure (II):
[0480] R.sup.30a and R.sup.30b are, at each occurrence,
independently H, --OH, --NH.sub.2, --CO.sub.2H, cyano, cyanoalkyl,
C.sub.1-C.sub.6alkyl, C.sub.3-C.sub.8cycloalkyl, hydroxylalkyl,
aminoalkyl, carboxylalkyl or aminocarbonyl;
[0481] R.sup.31a and R.sup.31b are, at each occurrence,
independently H, --OH, --NH.sub.2, --CO.sub.2H, cyano, cyanoalkyl,
C.sub.1-C.sub.6alkyl, C.sub.3-C.sub.8cycloalkyl, hydroxylalkyl,
aminoalkyl, carboxylalkyl or aminocarbonyl;
[0482] R.sup.32a and R.sup.32b are, at each occurrence,
independently H, --OH, --NH.sub.2, --CO.sub.2H, cyano, cyanoalkyl,
C.sub.1-C.sub.6alkyl, C.sub.3-C.sub.8cycloalkyl, hydroxylalkyl,
aminoalkyl, carboxylalkyl or aminocarbonyl; and
[0483] R.sup.33a and R.sup.33b are, at each occurrence,
independently H, --OH, --NH.sub.2, --CO.sub.2H, cyano, cyanoalkyl,
C.sub.1-C.sub.6alkyl, C.sub.3-C.sub.8cycloalkyl, hydroxylalkyl,
aminoalkyl, carboxylalkyl or aminocarbonyl.
[0484] In other embodiments, R.sup.30a, R.sup.3b, R.sup.31a,
R.sup.31b, R.sup.32a, R.sup.32b, R.sup.33a and R.sup.33b are
selected from H, C.sub.1-C.sub.6alkyl, hydroxylalkyl, cyano,
cyanoalkyl and aminocarbonyl, for example H, C.sub.1-C.sub.6alkyl,
hydroxylalkyl, cyano, and aminocarbonyl or in other embodiments H,
C.sub.1-C.sub.6alkyl and hydroxylalkyl.
[0485] In some of the foregoing embodiments, at least one of
R.sup.30a, R.sup.30b, R.sup.31a, R.sup.31bR.sup.32a, R.sup.32b,
R.sup.33a or R.sup.33b is H. For example, in some embodiments each
of R.sup.30a, R.sup.30b, R.sup.31a, R.sup.31b, R.sup.32a,
R.sup.32b, R.sup.33a or R.sup.33b is H.
[0486] In some other of the foregoing embodiments, at least one of
R.sup.3a, R.sup.30b, R.sup.31aR.sup.31b, R.sup.32a, R.sup.32b,
R.sup.33a or R.sup.33b is hydroxylalkyl.
[0487] In still other of the foregoing embodiments, at least one of
R.sup.30a, R.sup.30b, R.sup.31aR.sup.31b, R.sup.32a, R.sup.32b,
R.sup.33a or R.sup.33b is cyano.
[0488] In still more of the foregoing embodiments of compound (II),
at least one of R.sup.30a, R.sup.30b, R.sup.31a, R.sup.31b,
R.sup.32a, R.sup.32b, R.sup.33a Or R.sup.33b is aminocarbonyl.
[0489] In other embodiments, at least one of R.sup.30a, R.sup.30b,
R.sup.31a, R.sup.31b, R.sup.32a, R.sup.32b, R.sup.33a or R.sup.33b
is C.sub.1-C.sub.6alkyl.
[0490] In some embodiments, R.sup.30a and R.sup.30b join to form a
carbocyclic or heterocyclic ring. In different embodiments,
R.sup.31a and R.sup.31b join to form a carbocyclic or heterocyclic
ring. In more embodiments, R.sup.32a and R.sup.32b join to form a
carbocyclic or heterocyclic ring. In yet other embodiments,
R.sup.33a and R.sup.33b join to form a carbocyclic or heterocyclic
ring.
[0491] In even other embodiments, R.sup.30a is H, --OH, --NH.sub.2,
--CO.sub.2H, cyano, C.sub.1-C.sub.6alkyl,
C.sub.3-C.sub.8cycloalkyl, hydroxylalkyl, aminoalkyl, carboxylalkyl
or aminocarbonyl and R.sup.30b joins with R.sup.31b to form a
carbocyclic or heterocyclic ring.
[0492] In more embodiments, R.sup.31a is H, --OH, --NH.sub.2,
--CO.sub.2H, cyano, C.sub.1-C.sub.6alkyl,
C.sub.3-C.sub.8cycloalkyl, hydroxylalkyl, aminoalkyl, carboxylalkyl
or aminocarbonyl and R.sup.31b joins with R.sup.30b to form a
carbocyclic or heterocyclic ring.
[0493] In other embodiments, R.sup.32a is H, --OH, --NH.sub.2,
--CO.sub.2H, cyano, C.sub.1-C.sub.6alkyl,
C.sub.3-C.sub.8cycloalkyl, hydroxylalkyl, aminoalkyl, carboxylalkyl
or aminocarbonyl and R.sup.32b joins with R.sup.33b to form a
carbocyclic or heterocyclic ring.
[0494] In still more embodiments, R.sup.33a is H, --OH, --NH.sub.2,
--CO.sub.2H, cyano, C.sub.1-C.sub.6alkyl,
C.sub.3-C.sub.8cycloalkyl, hydroxylalkyl, aminoalkyl, carboxylalkyl
or aminocarbonyl and R.sup.33b joins with R.sup.32b to form a
carbocyclic or heterocyclic ring.
[0495] In some other embodiments, the compound is selected from a
compound in Table 2.
TABLE-US-00002 TABLE 2a Representative Compounds of Structure (II)
No. Structure Name II-1 ##STR00487## 1-(3-(4-(2-(4,5- dichloro-2-
hydroxyphenyl- amino)acetyl) piperazin-1- yl)azetidin-1-
yl)prop-2-en-1- one II-2 ##STR00488## 1-(3-(4-(2-(4,5- dichloro-2-
hydroxyphenyl- amino)acetyl)piperazin- 1-yl)pyrrolidin-1-
yl)prop-2-en-1- one II-3 ##STR00489## N-(1'-(2-(4,5- dichloro-2-
hydroxyphenyl- amino)acetyl)- 1,3'-biazetidin-3- yl)acrylamide II-4
##STR00490## 1-(4-(1-(2-(4,5- dichloro-2- hydroxyphenyl-
amino)acetyl)pyrrolidin- 3- yl)piperazin-1- yl)prop-2-en-1- one
II-5 ##STR00491## 1-(3-(4-(2-(2,4- dichloro-5- methoxyphenyl-
amino)acetyl) piperazin-1- yl)azetidin-1- yl)prop-2-en-1- one II-6
##STR00492## 1-(3-(4-(3-(4,5- dichloro-2- hydroxyphenyl)
propanoyl)piperazin- 1-yl)azetidin-1- yl)prop-2-en-1- one II-7
##STR00493## 1-(3-(4-(2-(4- chloro-2- hydroxy-5- methylphenyl-
amino)acetyl) piperazin-1- yl)pyrrolidin-1- yl)prop-2-en-1- one
II-8 ##STR00494## 1-(3-(4-(2-(5- chloro-2- hydroxy-4- methylphenyl-
amino)acetyl)piperazin- 1-yl)pyrrolidin- 1-yl)prop-2-en-1- one II-9
##STR00495## 5-(2-(4-(1- acryloylazetidin- 3-yl)piperazin-1- yl)-2-
oxoethylamino)- 2,4- dichlorobenzo- nitrile II-10 ##STR00496##
2-(2-(4-(1- acryloylazetidin- 3-yl)piperazin-1- yl)-2-
oxoethylamino)- 4,5- dichlorobenzamide II-11 ##STR00497##
1-(3-(4-(2-(4- chloro-2- hydroxy-5- iodophenylamino)
acetyl)piperazin- 1- yl)pyrrolidin-1- yl)prop-2-en-1- one II-12
##STR00498## 1-(3-(4-(2-(4- chloro-5-ethyl-2- hydroxyphenyl-
amino)acetyl)piperazin-1- yl)pyrrolidin-1- yl)prop-2-en-1- one
II-13 ##STR00499## 1-(3-(4-(2',5',6- trichloro-4- methoxybiphenyl-
carbonyl)piperazin- 1-yl)azetidin-1- yl)prop-2-en-1- one II-14
##STR00500## 1-(3-(4-(2-(5- chloro-4-fluoro-2- hydroxyphenyl-
amino)acetyl)piperazin-1- yl)azetidin- 1-yl)prop-2-en-1-one II-15
##STR00501## 1-(3-(4-(2-(4,5- dichloro-2- hydroxyphenyl-
amino)ethyl)piperazin- 1-yl)azetidin-1- yl)prop-2-en-1-one II-16
##STR00502## 4-(2-(4-(1- acryloylazetidin- 3-yl)piperazin-1-
yl)-2-oxoethyl)- 3,4- dihydroquinoxalin- 2(1H)-one II-17
##STR00503## 1-(1- acryloylazetidin- 3-yl)-N-(4,5- dichloro-2-
hydroxybenzyl) piperidine-4- carboxamide II-18 ##STR00504##
1-(3-(4-(2-(5- bromo-4-chloro- 2- hydroxyphenyl-
amino)acetyl)piperazin- 1-yl)azetidin- 1-yl)prop-2-en-1- one II-19
##STR00505## 1-(3-(4-(2-(5- chloro-2- hydroxyphenyl-
amino)acetyl)piperazin- 1-yl)azetidin-1- yl)prop-2-en-1- one II-20
##STR00506## (E)-1-(3-(4-((4,5- dichloro-2- hydroxyphenyl)
glycyl)piperazin-1- yl)azetidin-1-yl)- 4- (dimethylamino)
but-2-en-1-one II-21 ##STR00507## 1-(3-(4-(1- acryloylazetidin-
3-yl)piperazin-1- yl)-3- oxopropyl)-5,6- dichloro-1H-
benzo[d]imidazol- 2(3H)-one II-22 ##STR00508## 1-(3-(4-(2-(2,4,5-
trichlorophenyl- amino)acetyl)piperazin- 1-yl)azetidin-1-
yl)prop-2-en-1- one II-23 ##STR00509## 1-(3-(4-(2-(2,4- dichloro-5-
hydroxyphenyl- amino)acetyl)piperazin- 1-yl)azetidin-1-
yl)prop-2-en-1- one II-24 ##STR00510## 1-(3-(4-(2- (naphthalen-1-
yl)acetyl)piperazin- 1-yl)azetidin-1- yl)prop-2-en-1- one II-25
##STR00511## 1-(3-(4-(2-(1H- indol-3- yl)acetyl)piperazin-
1-yl)azetidin- 1-yl)prop-2-en- 1-one II-26 ##STR00512##
1-(3-(4-(2-(4,5- dichloro-2- (trifluoromethyl) phenylamino)
acetyl)piperazin-1- yl)azetidin-1- yl)prop-2-en-1- one II-27
##STR00513## 1-(3-(4-(2-(3,4- dichloro-5- hydroxyphenyl-
amino)acetyl) piperazin-1- yl)azetidin-1- yl)prop-2-en-1- one II-28
##STR00514## 1-(3-(4-(2-(4- bromo-5-chloro- 2- hydroxyphenyl-
amino)acetyl) piperazin-1-yl)azetidin- 1-yl)prop-2-en-1- one II-29
##STR00515## 1-(3-(4-(2-(1H- indol-1- yl)acetyl)piperazin-
1-yl)azetidin- 1-yl)prop-2-en- 1-one II-30 ##STR00516##
1-(3-(4-(2-(5,6- dichloro-1H- indol-1- yl)acetyl)piperazin-
1-yl)azetidin-1- yl)prop-2-en-1- one II-31 ##STR00517##
1-(3-(4-(2-(4,5- dichloro-2- hydroxyphenyl- amino)propanoyl)
piperazin-1- yl)azetidin-1- yl)prop-2-en-1- one II-32 ##STR00518##
1-(3-(4-(2-(4- chloro-2- hydroxy-5- methylphenyl-
amino)acetyl)piperazin- 1-yl)azetidin-1- yl)prop-2-en-1- one II-33
##STR00519## 1-(3-(4-(2-(3- chloro-5- hydroxyphenyl- amino)acetyl)
piperazin-1- yl)azetidin-1- yl)prop-2-en-1- one II-34 ##STR00520##
1-(3-(4-(2-(2- hydroxy-5- (methylsulfonyl) phenylamino)acetyl)
piperazin-1- yl)azetidin-1- yl)prop-2-en-1- one II-35 ##STR00521##
1-(3-(4-(2-(4- chloro-5- cyclopropyl-2- hydroxyphenyl)
amino)acetyl)piperazin-1- yl)azetidin-1- yl)prop-2-en-1- one II-36
##STR00522## N-(1'-(2-(5- bromo-4-chloro-2- hydroxyphenyl-
amino)acetyl)-1,3'- biazetidin-3- yl)acrylamide II-37 ##STR00523##
1-(3-(4-(2-(4- chloro-2- methoxy-5- (trifluoromethyl) phenylamino)
acetyl)piperazin-1- yl)azetidin-1- yl)prop-2-en-1- one II-38
##STR00524## 1-(3-(4-(2-(5- chlorothiazol-2- ylamino)acetyl)
piperazin-1- yl)azetidin-1- yl)prop-2-en-1- one II-39 ##STR00525##
1-(4-(1-(2-(4,5- dichloro-2- hydroxyphenyl- amino)acetyl)
azetidin-3- yl)piperazin-1- yl)prop-2-en-1- one II-40 ##STR00526##
2-(4,5-dichloro-2- hydroxyphenyl- amino)-1-(3-(4- (vinylsulfonyl)
piperazin-1- yl)azetidin-1- yl)ethanone II-41 ##STR00527##
2-(4,5-dichloro- 2- hydroxyphenyl- amino)-1-(4-(1- (vinylsulfonyl)
pyrrolidin-3- yl)piperazin-1- yl)ethanone II-42 ##STR00528##
4-(2-(4-(1- acryloylazetidin- 3-yl)piperazin-1- yl)-2-oxoethyl)-
3,4- dihydroquinoxalin- 2(1H)-one II-43 ##STR00529## 1-(3-(4-(2-(3-
hydroxynaphthalen- 2-ylamino)acetyl) piperazin-1- yl)azetidin-1-
yl)prop-2-en-1- one II-44 ##STR00530## 5-(2-(4-(1-
acryloylazetidin- 3-yl)piperazin-1- yl)-2- oxoethylamino)- 2,3-
dichlorobenzamide II-45 ##STR00531## N-(1'-(2-(4- chloro-5-
cyclopropyl-2- hydroxyphenyl- amino)acetyl)- 1,3'-biazetidin-3-
yl)acrylamide II-46 ##STR00532## 5-(2-(4-(1- acryloylazetidin-
3-yl)piperazin-1- yl)-2- oxoethylamino)- 2-chloro-4-
methoxybenzaldehyde II-47 ##STR00533## 1-(3-(4-(4- chloro-5-
cyclopropyl-2- methoxybenzoyl) piperazin-1- yl)azetidin-1-
yl)prop-2-en-1- one II-48 ##STR00534## 1-(3-(4-(2',5',6-
trichloro-4- hydroxybiphenyl- carbonyl)piperazin- 1-yl)azetidin-1-
yl)prop-2-en-1- one II-49 ##STR00535## 1-(4-(1- acryloylazetidin-
3-yl)piperazin-1- yl)-2-(4,5- dichloro-2- hydroxyphenyl-
amino)butan-1- one II-50 ##STR00536## 1-(3-(4-(2-(4- chloro-2-
hydroxy-5- isopropylphenyl- amino)acetyl) piperazin-1-
yl)azetidin-1- yl)prop-2-en-1- one II-51 ##STR00537## 1-(1-
acryloylazetidin- 3-yl)-4-(2-(4- chloro-5- cyclopropyl-2-
hydroxyphenyl- amino)acetyl) piperazine-2- carboxamide II-52
##STR00538## 1-(3-(4-(2-(5- chloro-4-ethyl-2- hydroxyphenyl-
amino)acetyl)piperazin- 1-yl)azetidin- 1-yl)prop-2-en-1- one II-53
##STR00539## 1-(3-(4-(2-(4- chloro-5- cyclobutyl-2- hydroxyphenyl-
amino)acetyl) piperazin-1- yl)azetidin-1- yl)prop-2-en-1- one II-54
##STR00540## 1-(3-(4-(2-(4- chloro-5- cyclopropyl-2- hydroxyphenyl-
amino)acetyl)-2- (hydroxymethyl) piperazin-1- yl)azetidin-1-
yl)prop-2-en-1- one II-55 ##STR00541## 1-(3-(4-(2-(4-
chloro-5-ethyl-2- hydroxyphenyl- amino)acetyl) piperazin-1-
yl)azetidin-1- yl)prop-2-en-1- one II-56 ##STR00542##
1-(3-(4-(2-(4- chloro-5-(2,2- difluorocyclopropyl)-2-
hydroxyphenyl- amino)acetyl) piperazin-1-yl)azetidin-
1-yl)prop-2-en-1- one II-57 ##STR00543## 1-(3-(4-(2-(4- chloro-2-
hydroxy-5- (2,2,2- trifluoroethyl) phenylamino) acetyl)piperazin-1-
yl)azetidin-1- yl)prop-2-en-1- one II-58 ##STR00544##
1-(3-(4-(3-(4- chloro-5- cyclopropyl-2- hydroxyphenyl)
propanoyl)piperazin- 1-yl)azetidin-1- yl)prop-2-en-1- one II-59
##STR00545## 1-(3-(4-(2-(4- chloro-5- cyclobutyl-2- hydroxyphenyl-
amino)propanoyl) piperazin-1- yl)azetidin-1- yl)prop-2-en-1- one
II-60 ##STR00546## 1-(3-(4-(2-(4- chloro-5- cyclopropyl-2-
hydroxyphenyl- amino)acetyl)-2- methylpiperazin- 1-yl)azetidin-1-
yl)prop-2-en-1- one II-61 ##STR00547## 1-(3-(4-(2-(5,6-
dichloro-1H- indol-3- yl)acetyl)piperazin- 1-yl)azetidin-1-yl)
prop-2-en-1-one II-62 ##STR00548## (E)-1-(4-(1- acryloylazetidin-
3-yl)piperazin-1- yl)-3-(4-chloro-5- cyclopropyl-2- hydroxyphenyl)
prop-2-en-1-one II-63 ##STR00549## (S)-1-(3-(4-(2- (4-chloro-5-
cyclopropyl-2- hydroxyphenyl- amino)acetyl)-2- (hydroxymethyl)
piperazin-1- yl)azetidin-1- yl)prop-2-en-1- one II-64 ##STR00550##
1-(3-(4-(2-(4- chloro-2- hydroxy-5-(1- methylcyclopropyl)
phenylamino) acetyl)piperazin-1- yl)azetidin-1- yl)prop-2-en-1- one
II-65 ##STR00551## 1-(3-(4-(2-(4- chloro-5- cyclopropyl-2-
methoxyphenyl- thio)acetyl)piperazin- 1-yl)azetidin-1-
yl)prop-2-en-1- one II-66 ##STR00552## 4-(1- acryloylazetidin-
3-yl)-N-(5- bromo-4-chloro- 2- hydroxybenzyl) piperazine-1-
carboxamide II-67 ##STR00553## (S)-1-(3-(4-(2- (4-chloro-5-
cyclopropyl-2- hydroxyphenyl- amino)propanoyl) piperazin-1-
yl)azetidin-1- yl)prop-2-en-1- one II-68 ##STR00554##
(R)-1-(3-(4-(2-(4- chloro-5- cyclopropyl-2- hydroxyphenyl-
amino)propanoyl) piperazin-1- yl)azetidin-1- yl)prop-2-en-1- one
II-69 ##STR00555## (S)-1-(3-(4-(2- (4-chloro-5- ethyl-2-
hydroxyphenyl- amino)propanoyl) piperazin-1- yl)azetidin-1-
yl)prop-2-en-1- one II-70 ##STR00556## (R)-1-(3-(4-(2-(4-
chloro-5-ethyl-2- hydroxyphenyl- amino)propanoyl) piperazin-1-
yl)azetidin-1- yl)prop-2-en-1- one II-71 ##STR00557## 2-(2-(4-(1-
acryloylazetidin- 3-yl)piperazin-1- yl)-2- oxoethylamino)-
5-chloro-4- cyclopropyl benzonitrile II-72 ##STR00558##
1-(3-(4-(3-(4- chloro-5-ethyl-2- hydroxyphenyl)- 1H-pyrazol-5-
yl)piperazin-1- yl)azetidin-1- yl)prop-2-en-1- one II-73
##STR00559## 1-(3-(4-(1-(4- chloro-5- cyclopropyl-2- methoxyphenyl)
pyrrolidine-2- carbonyl)piperazin-1- yl)azetidin-1-yl) prop-2-en-
1-one II-74 ##STR00560## 1-(3-(4-(2-(5- chloro-4- ethylpyridin-2-
ylamino)acetyl) piperazin-1- yl)azetidin-1- yl)prop-2-en-1- one
II-75 ##STR00561## 1-(3-(4-(2-(4,5- dichloro-7- methoxy-1H-
indol-1- yl)acetyl)piperazin- 1-yl)azetidin- 1-yl)prop-2-en- 1-one
II-76 ##STR00562## 1-(3-(4-(1-(4- chloro-5-ethyl-2- methoxyphenyl)
piperidin-3- yl)piperazin-1- yl)azetidin-1- yl)prop-2-en-1- one
[0496] Compounds of structure II are prepared according to
procedures well-known or derivable by one of ordinary skill in the
art, for example by procedures analogous to those exemplified in
the examples provided below. Each of the compounds in Table 2a was
prepared in such a manner and analyzed by mass spectrometry and/or
.sup.1H NMR. The mass spectrum ([M+H.sup.+] or [M+Na.sup.+]) and/or
NMR spectrum was found to be consistent with the structures in
Table 2a. Mass spectrometry data for the compounds in Table 2a are
provided in Table 2b.
TABLE-US-00003 TABLE 2b Experimental Mass Spectral Data for
Compounds in Table 2a [M + No. H.sup.+] No. [M + H.sup.+] No. [M +
H.sup.+] No. [M + H.sup.+] II-1 411.30 II-2 425.25 II-3 399.20 II-4
427.30 II-5 449.25* II-6 410.25 II-7 429.35* II-8 407.35 II-9
422.25 II-10 462.25* II-11 519.25 II-12 443.30* II-13 532.25* II-14
395.30 II-15 399.25 II-16 419.25 II-17 434.25* II-18 457.35 II-19
379.30 II-20 470.35 II-21 450.35 II-22 433.05 II-23 435.25* II-24
386.25* II-25 351.35 II-26 487.30* II-27 413.30 II-28 479.20* II-29
353.30 II-30 421.30 II-31 449.25* II-32 393.30 II-33 377.30.sup.+
II-34 423.35 II-35 441.30* II-36 445.20 II-37 461.30 II-38
368.30.sup.+ II-39 411.20 II-40 447.25.sup.+ II-41 463.20 II-42
382.40.sup.+ II-43 417.35* II-44 440.30 II-45 405.35 II-46 421.30
II-47 404.35 II-48 494.30 II-49 441.30 II-50 421.35 II-51 462.45
II-52 407.40 II-53 433.40 II-54 449.35 II-55 407.30 II-56 455.20
II-57 461.40 II-58 418.40 II-59 447.40 II-60 433.45 II-61 421.25
II-62 416.35 II-63 449.40 II-64 433.35 II-65 451.30 II-66 459.25
II-67 433.20 II-68 433.40 II-69 421.35 II-70 421.35 II-71 428.35
II-72 416.35 II-73 473.90 II-74 392.30 II-75 451.30 II-76 447.85
*[M + Na].sup.+ .sup.+[M - H].sup.-
[0497] General Reaction Scheme 18 illustrates an exemplary
procedure for preparing compounds of structure (II).
##STR00563##
[0498] Referring to General Reaction Scheme I, (II') and (II'') are
available from commercial sources and/or are easily prepared
according to procedures known in the art. All variables on (II')
and (II''), with the exception of M.sup.1, are as defined above. In
some procedures, M.sup.1 is NH. Briefly, an appropriately
substituted acid (II') is activated and reacted with an
appropriately substituted heterocycle (II'') under appropriate
coupling conditions. The L.sup.2-E moiety may be present in (II'')
as illustrated or may be installed after coupling For example
L.sup.2-E may be installed before or after coupling via acylation
(or thioacylation) using a reagent such as an acid chloride or
thionyl chloride.
[0499] It should be noted that variations of the above procedure
are possible, some of which are exemplified in the examples. For
example, in some procedures (II'') is monocyclic and the second
cyclic moiety is added after the coupling step. In other
procedures, the acid moiety is present on the cyclic moiety (II'')
and R.sup.1 is appropriately substituted with a nucleophilic moiety
to enable coupling to form (IIa).
[0500] Various other options are available to one of ordinary skill
in the art to add various substituents and or modify or reorder the
above described steps to arrive at different embodiments of
compounds of structure II. It should also be noted that various
substitutions on (II') and/or (II'') can be present during the
coupling step (in protected or unprotected form) or the
substituents can be added after (II') and (II'') are coupled.
Methods for inclusion of these substituents are known in the
art.
[0501] It is understood that although an exemplary procedure for
prepare (IIa) is provided above, other compounds of structure (II)
can be prepared by analogous methods. For example, the carbonyl of
(IIa) may be reduced to form compounds of structure (II) wherein
L.sup.1 does not comprise a carbonyl. Embodiments wherein L.sup.1
is heterocycloalkylene or heteroarylene can be prepared from
analogous methods, for example by use of Buchwald chemistry to
include the heterocycloalkylene or heteroarylene portion. Other
methods for preparation of different compounds of structure (II)
are known in the art.
[0502] Briefly, an appropriately substituted acid is reacted with
an appropriately substituted heterocycle under amide coupling
conditions. Acylation (or thioacylation) using a reagent such as an
acid chloride or thionyl chloride results in compounds of structure
(II). Various options are available to one of ordinary skill in the
art to add various substituents and/or modify or reorder the above
described steps to arrive at different embodiments of compounds of
structure (II). The appropriate acid is purchased commercially or
made according to well-known procedures.
3. Compounds of Structure (III)
[0503] In still other embodiments, the compound used in combination
with one or more additional therapeutic agent has the following
structure (III)
##STR00564##
or a pharmaceutically acceptable salt, tautomer, stereoisomer or
prodrug thereof, wherein:
[0504] A is CR.sup.37b, N or NR.sup.38a;
[0505] B is CR.sup.37c, N, NR.sup.38b or S
[0506] C is CR.sup.37d, N, NR.sup.38c Or S
[0507] G.sup.3 and G.sup.4 are each independently N or CR, wherein
R is H, cyano, halo or C.sup.1-C.sup.6alkyl;
[0508] L.sup.1a is a bond, --NH--, alkylene or heteroalkylene
[0509] L.sup.2 is a bond or alkylene;
[0510] R.sup.32a and R.sup.32b are, at each occurrence,
independently H, --OH, --NH.sub.2, --CO.sub.2H, cyano, cyanoalkyl,
C.sub.1-C.sub.6alkyl, C.sub.3-C.sub.8cycloalkyl, hydroxylalkyl,
aminoalkyl, carboxylalkyl or aminocarbonyl; or R.sup.32a and
R.sup.32b join to form a carbocyclic or heterocyclic ring; or
R.sup.32a is H, --OH, --NH.sub.2, --CO.sub.2H, cyano,
C.sub.1-C.sub.6alkyl, C.sub.3-C.sub.8cycloalkyl, hydroxylalkyl,
aminoalkyl, carboxylalkyl or aminocarbonyl and R.sup.32b joins with
R.sup.33b to form a carbocyclic or heterocyclic ring;
[0511] R.sup.33a and R.sup.33b are, at each occurrence,
independently H, --OH, --NH.sub.2, --CO.sub.2H, cyano, cyanoalkyl,
C.sub.1-C.sub.6alkyl, C.sub.3-C.sub.8cycloalkyl, hydroxylalkyl,
aminoalkyl, carboxylalkyl or aminocarbonyl; or R.sup.33a and
R.sup.33b join to form a carbocyclic or heterocyclic ring; or
R.sup.33a is H, --OH, --NH.sub.2, --CO.sub.2H, cyano,
C.sub.1-C.sub.6alkyl, C.sub.3-C.sub.8cycloalkyl, hydroxylalkyl,
aminoalkyl, carboxylalkyl or aminocarbonyl and R.sup.33b joins with
R.sup.32b to form a carbocyclic or heterocyclic ring;
[0512] R.sup.37a, R.sup.37b, R.sup.37c, R.sup.37d and R.sup.37e are
each independently H, halo, oxo, hydroxyl, cyano, aminocarbonyl,
formyl, C.sub.1-C.sub.6alkyl, C.sub.1-C.sub.6alkylsulfonyl,
C.sub.1-C.sub.6haloalkyl, C.sub.3-C.sub.8cycloalkyl,
C.sub.1-C.sub.6alkoxy, C.sub.1-C.sub.6hydroxylalkyl,
C.sub.1-C.sub.6alkoxyalkyl, C.sub.1-C.sub.6aminoalkyl, heterocyclyl
or aryl;
[0513] R.sup.38a, R.sup.38b and R.sup.38c are each independently H,
C.sub.1-C.sub.6alkyl or aryl;
[0514] n.sup.3 and n.sup.4 are each independently 1, 2 or 3
[0515] m is 0 or 1;
[0516] is a single or double bond such that all valences are
satisfied; and
[0517] E is an electrophilic moiety capable of forming a covalent
bond with the cysteine residue at position 12 of a KRAS, HRAS or
NRAS G12C mutant protein.
[0518] In various other embodiments, the compound has one of the
following structures (IIIa), (IIIb), (IIIc), (IIId), (IIIe), (IIIf)
or (IIIg):
##STR00565##
[0519] In some different embodiments, the compound has one of the
following structures (IIIa'), (IIIb'), (IIIc'), (IIId'), (IIIe'),
(IIIf') or (IIIg'):
##STR00566## ##STR00567##
wherein:
[0520] Q is --C(.dbd.O)--, --NR.sup.34C(.dbd.O)--,
--S(.dbd.O).sub.2-- or --NR.sup.34S(.dbd.O).sub.2--;
[0521] R.sup.34 is H, C.sub.1-C.sub.6alkyl or hydroxylalkyl;
[0522] is a carbon-carbon double bond or a carbon-carbon triple
bond; and
[0523] R.sup.35 and R.sup.36 are each independently H, cyano,
C.sub.1-C.sub.6alkyl, aminoalkyl, alkylaminoalkyl, or hydroxylalkyl
or R.sup.35 and R.sup.36 join to form a carbocyclic or heterocyclic
ring when is a double bond; or R.sup.35 is absent and R.sup.36 is
H, C.sub.1-C.sub.6alkyl, aminoalkyl, alkylaminoalkyl or
hydroxylalkyl when is a triple bond.
[0524] In some specific embodiments of the foregoing compounds of
structure (III), and substructures thereof, R.sup.37a is halo, aryl
or heteroaryl. In further such embodiments, R.sup.35 and R.sup.36
are each H.
[0525] In various other embodiments, G.sup.3 is N and G.sup.4 is
CR, for example CH.
[0526] In some different embodiments, G.sup.3 is CR, for example,
CH, and G.sup.4 is N.
[0527] In still other embodiments, G.sup.3 is N and G.sup.4 is
N.
[0528] In various other embodiments, n.sup.3 is 2 and n.sup.4 is 2.
In still other embodiments, n.sup.3 is 1 and n.sup.4 is 1. In some
more embodiments, n.sup.3 is 2 and n.sup.4 is 1.
[0529] In other of the foregoing embodiments, R.sup.37a, R.sup.37b,
R.sup.37c, R.sup.37d and R.sup.37e are each independently H, --OH,
halo, oxo, C.sub.1-C.sub.6alkyl, C.sub.1-C.sub.6alkoxy,
heterocyclyl or aryl.
[0530] In still other embodiments, R.sup.37a, R.sup.37b, R.sup.37c,
R.sup.37d and R.sup.37e are each independently H, --OH, fluoro,
chloro, bromo, iodo, oxo, methyl, methoxy, heteroaryl or aryl.
[0531] In some embodiments, R.sup.37a or R.sup.37e is aryl. In some
more specific embodiments, R.sup.37a is aryl, such as phenyl.
[0532] In some different embodiments, the aryl is unsubstituted. In
some other embodiments, the aryl is substituted. For example, in
some embodiments the aryl is substituted with one or more halo
substituents. In some of these embodiments, the halo substituents
are selected from fluoro and chloro.
[0533] In still other embodiments, R.sup.37a is heteroaryl. In some
of these embodiments, the heteroaryl is unsubstituted. In various
other embodiments, the heteroaryl is substituted. In some more
embodiments, the heteroaryl comprises nitrogen, sulfur or a
combination thereof.
[0534] In some more specific embodiments, the heteroaryl is
thiophenyl.
[0535] In other of the foregoing embodiments, R.sup.37a is halo.
For example, in some embodiments halo is chloro, bromo or iodo.
[0536] In some embodiments, R.sup.37a or R.sup.37e has one of the
following structures:
##STR00568##
[0537] In still other embodiments, R.sup.37a has one of the
following structures:
##STR00569##
[0538] In various different embodiments, R.sup.38a, R.sup.38b and
R.sup.38c are each independently H or aryl. In still other
embodiments, R.sup.38a, R.sup.38b and R.sup.38c are each
independently H.
[0539] In some other different embodiments, R.sup.38c is aryl. For
example, in some embodiments the aryl is substituted with one or
more halo substituents. In some of these embodiments, halo is
chloro.
[0540] In some other embodiments of the compounds of structure
(III), Q is --C(.dbd.O)--. In some other embodiments, Q is
--S(.dbd.O).sub.2--. In still other embodiments, Q is
--NR.sup.34C(.dbd.O)--. In still more other embodiments, Q is
--NR.sup.34S(.dbd.O).sub.2--.
[0541] In some more specific embodiments, R.sup.34 is H. For
example, in some embodiments R.sup.34 is hydroxylalkyl, such as
2-hydroxylalkyl.
[0542] In other of the foregoing embodiments, at least one of
R.sup.35 or R.sup.36 is H. For example, in some embodiment search
of R.sup.35 and R.sup.36 are H.
[0543] In various other embodiments, R.sup.36 is alkylaminoalkyl.
For example, in some embodiments R.sup.36 has the following
structure:
##STR00570##
[0544] In some different embodiments, R.sup.36 is hydroxylalkyl,
for example 2-hydroxylalkyl In various other embodiments, R.sup.35
and R.sup.36 join to form a ring. In some of these embodiments, the
ring is a cyclopentene, cyclohexene or phenyl ring.
[0545] In other of the foregoing embodiments, E has one of the
following structures:
##STR00571##
[0546] In some embodiments, E is
##STR00572##
[0547] In some more of the foregoing embodiments, L.sup.1 is
heteroalkylene. In some more embodiments, the heteroalkylene is
unsubstituted. In some different embodiments, the heteroalkylene is
substituted.
[0548] In various other embodiments, L is aminoalkylene. For
example, in some embodiments L.sup.1 is --CH.sub.2CH.sub.2NH--.
[0549] In some different embodiments, L.sup.1a is a bond.
[0550] In some embodiments, L.sup.1a is alkylene, alkenylene,
heteroalkylene or heterocycloalkylene. In some other embodiments,
L.sup.1a is alkylene or heteroalkylene. In some of these
embodiments, L.sup.1a is substituted alkylene. In various other
embodiments, L.sup.1a is unsubstituted alkylene. For example, in
some embodiments L.sup.1a is
##STR00573##
[0551] In some different embodiments, L.sup.1a is substituted
heteroalkylene. In some other embodiments, L.sup.1a is
unsubstituted heteroalkylene. In some of the foregoing embodiments,
L.sup.1a is aminoalkylene or thioalkylene, for example
aminoalkylene. For example, in some embodiments L.sup.1a has one of
the following structures:
##STR00574##
[0552] In other embodiments, L.sup.1a is
##STR00575##
[0553] In other embodiments, L.sup.1a is substituted alkenylene. In
different embodiments, L.sup.1a is unsubstituted alkenylene. In
some more specific embodiments, L.sup.1a has the following
structure:
##STR00576##
[0554] In yet other embodiments, L.sup.1a is substituted
heterocycloalkylene. In some other embodiments, L.sup.1a is
unsubstituted heterocycloalkylene. For Example, in some
embodiments, L.sup.1a has the following structure:
##STR00577##
[0555] In some of the foregoing embodiments, L.sup.2 is a bond.
[0556] In various other embodiments, L.sup.2 is substituted
alkylene. In still other embodiments, L.sup.2 is unsubstituted
alkylene.
[0557] In some embodiments of any of the foregoing compounds of
structure (III):
[0558] R.sup.32a and R.sup.32b are, at each occurrence,
independently H, --OH, --NH.sub.2, --CO.sub.2H, cyano, cyanoalkyl,
C.sub.1-C.sub.6alkyl, C.sub.3-C.sub.8cycloalkyl, hydroxylalkyl,
aminoalkyl, carboxylalkyl or aminocarbonyl; and
[0559] R.sup.33a and R.sup.33b are, at each occurrence,
independently H, --OH, --NH.sub.2, --CO.sub.2H, cyano, cyanoalkyl,
C.sub.1-C.sub.6alkyl, C.sub.3-C.sub.8cycloalkyl, hydroxylalkyl,
aminoalkyl, carboxylalkyl or aminocarbonyl.
[0560] In other embodiments, R.sup.32a, R.sup.32b, R.sup.33a or
R.sup.33b are selected from H, C.sub.1-C.sub.8alkyl, hydroxylalkyl,
cyano, cyanoalkyl and aminocarbonyl, for example H, hydroxyl alkyl
and cyano.
[0561] In other of the foregoing embodiments, at least one of
R.sup.32a, R.sup.32b, R.sup.33a or R.sup.33b is H. For example, in
some embodiments each of R.sup.32a, R.sup.32b, R.sup.33a or
R.sup.33b is H.
[0562] In other of the foregoing embodiments, at least one of
R.sup.32a, R.sup.32b, R.sup.33a or R.sup.33b is hydroxylalkyl.
[0563] In still other embodiments, at least one of R.sup.32a,
R.sup.32b, R.sup.33a or R.sup.33b is cyano.
[0564] In some other different embodiments, least one of R.sup.32a,
R.sup.32b, R.sup.33a or R.sup.33b is aminocarbonyl.
[0565] In some embodiments, R.sup.32a and R.sup.32b join to form a
carbocyclic or heterocyclic ring. In other embodiments, R.sup.33a
and R.sup.33b join to form a carbocyclic or heterocyclic ring.
[0566] In different embodiments, R.sup.32a is H, --OH, --NH.sub.2,
--CO.sub.2H, cyano, C.sub.1-C.sub.6alkyl,
C.sub.3-C.sub.8cycloalkyl, hydroxylalkyl, aminoalkyl, carboxylalkyl
or aminocarbonyl and R.sup.32b joins with R.sup.33b to form a
carbocyclic or heterocyclic ring.
[0567] In still other embodiments, R.sup.33a is H, --OH,
--NH.sub.2, --CO.sub.2H, cyano, C.sub.1-C.sub.6alkyl,
C.sub.3-C.sub.8cycloalkyl, hydroxylalkyl, aminoalkyl, carboxylalkyl
or aminocarbonyl and R.sup.33b joins with R.sup.32b to form a
carbocyclic or heterocyclic ring.
[0568] In some more specific embodiments, the compound of structure
(III) is selected from a compound in Table 3.
TABLE-US-00004 TABLE 3 Representative Compounds of Structure (III)
No. Structure Name [M + H].sup.+ VI-1 ##STR00578##
1-(4-(2',6-dichloro-4- methoxybiphenylcarbonyl) piperazin-1-
yl)prop-2-en-1-one 441.20* VI-2 ##STR00579## 1-(4-(4-chloro-5-iodo-
2- methoxybenzoyl) piperazin-1-yl)prop-2-en-1- one 457.05* VI-3
##STR00580## 1-(4-(2',6-dichloro-4- hydroxybiphenyl-
carbonyl)piperazin-1- yl)prop-2-en-1-one 427.15 VI-4 ##STR00581##
1-(4-2',6-dichloro-5'- fluoro-4- methoxybiphenylcarbonyl)
piperazin-1- yl)prop-2-en-1-one 459.15 VI-5 ##STR00582##
1-(4-(2',5',6-trichloro- 4- methoxybiphenylcarbonyl) piperazin-1-
yl)prop-2-en-1-one 453.15 VI-6 ##STR00583## 1-(2-(hydroxymethyl)-
4-(2',5',6-trichloro-4- methoxybiphenylcarbonyl) piperazin-1-
yl)prop-2-en-1-one 483.20.sup.# VI-7 ##STR00584##
1-(3-(hydroxymethyl)- 4-(2',5',6-trichloro-4-
methoxybiphenylcarbonyl) piperazin-1- yl)prop-2-en-1-one 485.20
VI-8 ##STR00585## 1-(4-(4,5-dichloro-2- hydroxybenzoyl)piperazin-1-
yl)prop-2-en-1-one 327.15 VI-9 ##STR00586## 1-(4-(5-bromo-4-
chloro-2- hydroxybenzoyl)piperazin-1- yl)prop-2-en-1-one 388* VI-10
##STR00587## (E)-1-(4-(2',6- dichloro-4- methoxybiphenylcarbonyl)
piperazin-1-yl)-4- (dimethylamino) but-2-en-1-one 476.23 VI-11
##STR00588## 1-(4-(3-(2- chlorophenyl)-1H- pyrazole-5-
carbonyl)piperazin-1- yl)prop-2-en-1-one 467.20* VI-12 ##STR00589##
1-(4-(2',6- dichlorobiphenylcarbonyl) piperazin-1-
yl)prop-2-en-1-one 389.20 VI-13 ##STR00590## N-(1-(2',6-dichloro-4-
hydroxybiphenyl- carbonyl)azetidin-3- yl)acrylamide 389.30 VI-14
##STR00591## N-(1-acryloylazetidin- 3-yl)-3-(2-chlorophenyl)-1H-
pyrazole-5- carboxamide 331.15 VI-15 ##STR00592## N-(1-(3-(2-
chlorophenyl)-1H- pyrazole-5- carbonyl)azetidin-3- yl)acrylamide
331.15 VI-16 ##STR00593## 1-(4-(5-(thiophen-2- yl)-1H-pyrazole-3-
carbonyl)piperazin-1- yl)prop-2-en-1-one 339.15 VI-17 ##STR00594##
1-(4-(2',5',6-trichloro- 4- hydroxybiphenylcarbonyl) piperazin-1-
yl)prop-2-en-1-one .sup. 438.30.sup.+ VI-18 ##STR00595##
1-(4-(2-(2- chlorophenyl)thiazole- 4-carbonyl)piperazin-
1-yl)prop-2-en-1-one .sup. 384.15.sup.+ VI-19 ##STR00596##
1-(4-(4-(2- chlorophenyl)thiazole- 2-carbonyl)piperazin-
1-yl)prop-2-en-1-one .sup. 384.15.sup.+ VI-20 ##STR00597##
1-(4-(5-chloro-2-(2- chlorophenyl)thiazole- 4-carbonyl)piperazin-
1-yl)prop-2-en-1-one 418.10* VI-21 ##STR00598## 1-(4-(4-(2-
chlorophenyl)-1H- pyrrole-2- carbonyl)piperazin-1-
yl)prop-2-en-1-one 344.20 VI-22 ##STR00599## 1-(4-(4-(2-
chlorophenyl)-5- methylthiazole-2- carbonyl)piperazin-1-
yl)prop-2-en-1-one 376.25 VI-23 ##STR00600## 1-(4-(4-(2-
chlorophenyl)-5- methyl-1H-pyrrole-2- carbonyl)piperazin-1-
yl)prop-2-en-1-one .sup. 356.35.sup.+ VI-24 ##STR00601##
(E)-1-(4-(2',6- dichloro-4- hydroxybiphenylcarbonyl)
piperazin-1-yl)-4- (dimethylamino)but- 2-en-1-one 462.35 VI-25
##STR00602## 1-(4-(2'-chloro-5- hydroxybiphenylcarbonyl)
piperazin-1- yl)prop-2-en-1-one 371.25 VI-26 ##STR00603##
1-(4-(2'-chloro-4- hydroxybiphenyl- carbonyl)piperazin-
1-yl)prop-2-en-1-one 371.25 VI-27 ##STR00604##
1-(4-(2',6-dichloro-5- hydroxybiphenylcarbonyl) piperazin-1-
yl)prop-2-en-1-one .sup. 403.35.sup.+ VI-28 ##STR00605## 5-(4-
acryloylpiperazine-1- carbonyl)-1-(2,5- dichlorophenyl)-4-
hydroxypyridin- 2(1H)-one 422.06 VI-29 ##STR00606##
1-(4-(6-chloro-4- hydroxybiphenylcarbonyl)- piperazin-1-
yl)prop-2-en-1-one 371.25 VI-30 ##STR00607## 1-(4-(5-chloro-4-(2-
chlorophenyl)-1H- pyrrole-2- carbonyl)piperazin-1-
yl)prop-2-en-1-one .sup. 378.25.sup.+ VI-31 ##STR00608##
1-(4-(4-(2,5- dichlorophenyl)-1H- pyrrole-2- carbonyl)piperazin-1-
yl)prop-2-en-1-one 378.20 VI-32 ##STR00609## 1-(4-(4-(2,4-
dichlorophenyl)-1H- pyrrole-2- carbonyl)piperazin-1-
yl)prop-2-en-1-one 377.90 VI-33 ##STR00610## N-(1-(3,4-
dichlorobenzoyl) piperidin-4- yl)ethenesulfonamide 363.02 VI-34
##STR00611## (3,4- dichlorophenyl)(4- (vinylsulfonyl)piperazin-
1-yl)methanone 349.04 VI-35 ##STR00612## (S)-N-(1-(3,4-
dichlorobenzoyl) piperidin-3- yl)ethenesulfonamide 363.06 VI-36
##STR00613## 1-(4-(3,4- dichlorobenzoyl)
piperazin-1-yl)prop-2-en-1- one 312.99 VI-37 ##STR00614##
1-acryloyl-4-(4',6- dichloro-4- hydroxybiphenyl-
carbonyl)piperazine-2- carbonitrile 430.30 VI-38 ##STR00615##
1-acryloyl-4-(2',5',6- trichloro-4- hydroxybiphenyl-
carbonyl)piperazine-2- carbonitrile 464.30 VI-39 ##STR00616##
1-acryloyl-4-(4,5- dichloro-2- hydroxybenzoyl)
piperazine-2-carbonitrile 354.15 VI-40 ##STR00617##
1-(4-(2-chloro-5- hydroxybiphenylcarbonyl) piperazin-1-
yl)prop-2-en-1-one 371.25 VI-41 ##STR00618## 1-(4-(2,2'-dichloro-5-
hydroxybiphenylcarbonyl) piperazin-1- yl)prop-2-en-1-one 405.25
VI-42 ##STR00619## 1-(4-(2,4'-dichloro-5- hydroxybiphenylcarbonyl)
piperazin-1- yl)prop-2-en-1-one 405.20 VI-43 ##STR00620##
1-(4-(2,3'-dichloro-5- hydroxybiphenylcarbonyl) piperazin-1-
yl)prop-2-en-1-one 405.25 *[M + Na].sup.+ .sup.+[M - H].sup.-
.sup.#[M]
[0569] Compounds of structure III are prepared according to
procedures well-known or derivable by one of ordinary skill in the
art, for example by procedures analogous to those exemplified in
the examples provided below. Each of the compounds in Table 3 was
prepared in such a manner and analyzed by mass spectrometry and/or
.sup.1H NMR. The mass spectrum ([M+H.sup.+] or [M+Na+]) and/or NMR
spectrum was found to be consistent with the structures in Table
3.
[0570] General Reaction Scheme 19 illustrates an exemplary
procedure for preparing compounds of structure (III).
##STR00621##
[0571] Referring to General Reaction Scheme 19, (III') and (III'')
are available from commercial sources and/or are easily prepared
according to procedures known in the art.
[0572] All variables on (III') and (III''), with the exception of
M.sup.1 and M.sup.2, are as defined above for compounds of
structure (III). In some procedures, M.sup.1 is NH and M.sup.2 is
absent. In other procedures M.sup.1 is N or CH and M.sup.2 is a
precursor to L.sup.1a which reacts with an activated acid. For
example, in various procedures M.sup.2 is NH.sub.2, aminoalkyl or
other heterosubstituted alkyl. Embodiments where M.sup.2 comprises
a carbanion (or M.sup.1 is a carbanion) are also contemplated such
that L.sup.1 is alkylene. Briefly, an appropriately substituted
acid (III') is activated and reacted with an appropriately
substituted heterocycle (III'') under appropriate coupling
conditions. The L.sup.2-E moiety may be present in (III'') as
illustrated or may be installed after coupling For example L2-E may
be installed before or after coupling via acylation (or
thioacylation) using a reagent such as an acid chloride or thionyl
chloride.
[0573] It should be noted that variations of the above procedure
are possible, some of which are exemplified in the examples. For
example, in some procedures, the acid moiety is present on the
cyclic moiety (III') and (III') is appropriately substituted with a
nucleophilic moiety to enable coupling to form (III). Other methods
of bond formation, which do not require reaction of an activated
acid, are also available for preparation of the compounds. It
should also be noted that various substitutions on (III') and/or
(III'') can be present during the coupling step (in protected or
unprotected form) or the substituents can be added after (III') and
(III'') are coupled. Methods for inclusion of these substituents
are known in the art.
[0574] Various options are available to one of ordinary skill in
the art to add various substituents and or modify or reorder the
above described steps to arrive at different embodiments of
compounds of structure III. The appropriate acid is purchased
commercially or made according to well-known procedures.
[0575] It will also be appreciated by those skilled in the art that
in the processes described herein (e.g., General Reaction Scheme I
and II and the below examples) the functional groups of
intermediate compounds may need to be protected by suitable
protecting groups. Such functional groups include hydroxy, amino,
mercapto and carboxylic acid. Suitable protecting groups for
hydroxy include trialkylsilyl or diarylalkylsilyl (for example,
t-butyldimethylsilyl, t-butyldiphenylsilyl or trimethylsilyl),
tetrahydropyranyl, benzyl, and the like. Suitable protecting groups
for amino, amidino and guanidino include t-butoxycarbonyl,
benzyloxycarbonyl, and the like. Suitable protecting groups for
mercapto include --C(O)--R'' (where R'' is alkyl, aryl or
arylalkyl), p-methoxybenzyl, trityl and the like. Suitable
protecting groups for carboxylic acid include alkyl, aryl or
arylalkyl esters. Protecting groups may be added or removed in
accordance with standard techniques, which are known to one skilled
in the art and as described herein.
[0576] The use of protecting groups is described in detail in
Green, T. W. and P. G. M. Wutz, Protective Groups in Organic
Synthesis (1999), 3rd Ed., Wiley. As one of skill in the art would
appreciate, the protecting group may also be a polymer resin such
as a Wang resin, Rink resin or a 2-chlorotrityl-chloride resin.
[0577] It will also be appreciated by those skilled in the art,
although such protected derivatives of compounds of this invention
may not possess pharmacological activity as such, they may be
administered to a subject and thereafter metabolized in the body to
form compounds of the invention which are pharmacologically active.
Such derivatives may therefore be described as "prodrugs". All
prodrugs of compounds of this invention are included within the
scope of the invention.
[0578] It is also understood that one skilled in the art may be
able to make these compounds by similar methods or by combining
other methods known to one skilled in the art. It is further
understood that one skilled in the art would be able to make, in a
similar manner as described below, other compounds of the invention
not specifically illustrated below by using the appropriate
starting components and modifying the parameters of the synthesis
as needed. In general, starting components may be obtained from
sources such as Sigma Aldrich, Lancaster Synthesis, Inc.,
Maybridge, Matrix Scientific, TCI, and Fluorochem USA, etc. or
synthesized according to sources known to those skilled in the art
(see, for example, Advanced Organic Chemistry: Reactions,
Mechanisms, and Structure, 5th edition (Wiley, December 2000)) or
prepared as described in this invention.
[0579] In some other embodiments, the compound has one of the
following structures:
##STR00622## ##STR00623##
C. Pharmaceutical Compositions
[0580] Other embodiments are directed to pharmaceutical
compositions. The pharmaceutical composition comprises any one (or
more) of the foregoing compounds, any one of the foregoing addition
therapeutic agents and a pharmaceutically acceptable carrier.
[0581] In some embodiments, the pharmaceutical composition is
formulated for oral administration. In other embodiments, the
pharmaceutical composition is formulated for injection.
[0582] Suitable routes of administration include, but are not
limited to, oral, intravenous, rectal, aerosol, parenteral,
ophthalmic, pulmonary, transmucosal, transdermal, vaginal, optical,
nasal, and topical administration. In addition, by way of example
only, parenteral delivery includes intramuscular, subcutaneous,
intravenous, intramedullary injections, as well as intrathecal,
direct intraventricular, intraperitoneal, intralymphatic, and
intranasal injections.
[0583] In certain embodiments, the compound and/or additional
therapeutic agent as described herein are administered in a local
rather than systemic manner, for example, via injection of the
compound and/or additional therapeutic agent directly into an
organ, often in a depot preparation or sustained release
formulation. In specific embodiments, long acting formulations are
administered by implantation (for example subcutaneously or
intramuscularly) or by intramuscular injection. Furthermore, in
other embodiments, the compound and additional therapeutic agent
are delivered in a targeted drug delivery system, for example, in a
liposome coated with organ-specific antibody. In such embodiments,
the liposomes are targeted to and taken up selectively by the
organ. In yet other embodiments, the compound and/or additional
therapeutic agent is provided in the form of a rapid release
formulation, in the form of an extended release formulation, or in
the form of an intermediate release formulation. In yet other
embodiments, the compound and/or additional therapeutic agent
described herein is administered topically.
[0584] The compound and additional therapeutic agent according to
the invention are effective over a wide dosage range. For example,
in the treatment of adult humans, dosages from 0.01 to 1000 mg,
from 0.5 to 100 mg, from 1 to 50 mg per day, and from 5 to 40 mg
per day are examples of dosages that are used in some embodiments.
An exemplary dosage is 10 to 30 mg per day. The exact dosage will
depend upon the route of administration, the form in which the
compound and additional therapeutic agent are administered, the
subject to be treated, the body weight of the subject to be
treated, and the preference and experience of the attending
physician.
[0585] In some embodiments, the compound and/or additional
therapeutic agent is administered in a single dose. Typically, such
administration will be by injection, e.g., intravenous injection,
in order to introduce the agent quickly. However, other routes are
used as appropriate. A single dose of the compound and/or
additional therapeutic agent of the invention may also be used for
treatment of an acute condition.
[0586] In some embodiments, the compound and/or additional
therapeutic agent of the invention is administered in multiple
doses. In some embodiments, dosing is about once, twice, three
times, four times, five times, six times, or more than six times
per day. In other embodiments, dosing is about once a month, once
every two weeks, once a week, or once every other day. In another
embodiment the compound and additional therapeutic agent are
administered together about once per day to about 6 times per day.
In another embodiment the administration of the compound and
additional therapeutic agent continues for less than about 7 days.
In yet another embodiment the administration continues for more
than about 6, 10, 14, 28 days, two months, six months, or one year.
In some cases, continuous dosing is achieved and maintained as long
as necessary.
[0587] Administration of the compound and additional therapeutic
agent may continue as long as necessary. In some embodiments, the
compound and/or additional therapeutic agent are administered for
more than 1, 2, 3, 4, 5, 6, 7, 14, or 28 days. In some embodiments,
the compound and additional therapeutic agent are administered for
less than 28, 14, 7, 6, 5, 4, 3, 2, or 1 day. In some embodiments,
the compound and additional therapeutic agent is administered
chronically on an ongoing basis, e.g., for the treatment of chronic
effects.
[0588] In some embodiments, the compound and/or additional
therapeutic agent are administered in dosages. It is known in the
art that due to intersubjective variability in compound
pharmacokinetics, individualization of dosing regimen is necessary
for optimal therapy. Dosing for a compound of the invention may be
found by routine experimentation in light of the instant
disclosure.
[0589] In some embodiments, the compound and/or additional
therapeutic agent described herein are formulated into
pharmaceutical compositions. In specific embodiments,
pharmaceutical compositions are formulated in a conventional manner
using one or more physiologically acceptable carriers comprising
excipients and auxiliaries which facilitate processing of the
active compounds into preparations which can be used
pharmaceutically. Proper formulation is dependent upon the route of
administration chosen. Any pharmaceutically acceptable techniques,
carriers, and excipients are used as suitable to formulate the
pharmaceutical compositions described herein: Remington: The
Science and Practice of Pharmacy, Nineteenth Ed (Easton, Pa.: Mack
Publishing Company, 1995); Hoover, John E., Remington's
Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa. 1975;
Liberman, H. A. and Lachman, L., Eds., Pharmaceutical Dosage Forms,
Marcel Decker, New York, N.Y., 1980; and Pharmaceutical Dosage
Forms and Drug Delivery Systems, Seventh Ed. (Lippincott Williams
& Wilkins 1999).
[0590] A pharmaceutical composition, as used herein, refers to a
mixture of compound and/or additional therapeutic agent with other
chemical components, such as carriers, stabilizers, diluents,
dispersing agents, suspending agents, thickening agents, and/or
excipients. In certain embodiments, the pharmaceutical composition
facilitates administration of the compound to an organism. In some
embodiments, practicing the methods of treatment or use provided
herein, therapeutically effective amounts of compounds of the
compound and/or additional therapeutic agent provided herein are
administered in a pharmaceutical composition to a subject having a
disease, disorder or medical condition to be treated. In specific
embodiments, the subject is a human. In certain embodiments,
therapeutically effective amounts vary depending on the severity of
the disease, the age and relative health of the subject, the
potency of the compound used and other factors.
[0591] In one embodiment, the compound and/or additional
therapeutic agent is formulated in an aqueous solution. In specific
embodiments, the aqueous solution is selected from, by way of
example only, a physiologically compatible buffer, such as Hank's
solution, Ringer's solution, or physiological saline buffer. In
other embodiments, compound and/or additional therapeutic agent are
formulated for transmucosal administration. In specific
embodiments, transmucosal formulations include penetrants that are
appropriate to the barrier to be permeated. In still other
embodiments wherein the compound and/or additional therapeutic
agent described herein are formulated for other parenteral
injections, appropriate formulations include aqueous or non-aqueous
solutions.
[0592] In specific embodiments, such solutions include
physiologically compatible buffers and/or excipients.
[0593] In another embodiment, the compound and/or additional
therapeutic agent described herein are formulated for oral
administration. The compound and/or additional therapeutic agent
described herein are formulated by combining the components with,
e.g., pharmaceutically acceptable carriers or excipients. In
various embodiments, the compound and/or additional therapeutic
agent described herein are formulated in oral dosage forms that
include, by way of example only, tablets, powders, pills, dragees,
capsules, liquids, gels, syrups, elixirs, slurries, suspensions and
the like.
[0594] In certain embodiments, pharmaceutical preparations for oral
use are obtained by mixing one or more solid excipient with the
compound and/or additional therapeutic agent described herein,
optionally grinding the resulting mixture, and processing the
mixture of granules, after adding suitable auxiliaries, if desired,
to obtain tablets or dragee cores. Suitable excipients are, in
particular, fillers such as sugars, including lactose, sucrose,
mannitol, or sorbitol; cellulose preparations such as: for example,
maize starch, wheat starch, rice starch, potato starch, gelatin,
gum tragacanth, methylcellulose, microcrystalline cellulose,
hydroxypropylmethylcellulose, sodium carboxymethylcellulose; or
others such as: polyvinylpyrrolidone (PVP or povidone) or calcium
phosphate. In specific embodiments, disintegrating agents are
optionally added.
[0595] Disintegrating agents include, by way of example only,
cross-linked croscarmellose sodium, polyvinylpyrrolidone, agar, or
alginic acid or a salt thereof such as sodium alginate.
[0596] In one embodiment, dosage forms, such as dragee cores and
tablets, are provided with one or more suitable coating. In
specific embodiments, concentrated sugar solutions are used for
coating the dosage form. The sugar solutions optionally contain
additional components, such as by way of example only, gum arabic,
talc, polyvinylpyrrolidone, carbopol gel, polyethylene glycol,
and/or titanium dioxide, lacquer solutions, and suitable organic
solvents or solvent mixtures. Dyestuffs and/or pigments are also
optionally added to the coatings for identification purposes.
Additionally, the dyestuffs and/or pigments are optionally utilized
to characterize different combinations of active compound
doses.
[0597] In certain embodiments, therapeutically effective amounts of
the compound and/or additional therapeutic agent described herein
are formulated into other oral dosage forms. Oral dosage forms
include push-fit capsules made of gelatin, as well as soft, sealed
capsules made of gelatin and a plasticizer, such as glycerol or
sorbitol. In specific embodiments, push-fit capsules contain the
active ingredients in admixture with one or more filler. Fillers
include, by way of example only, lactose, binders such as starches,
and/or lubricants such as talc or magnesium stearate and,
optionally, stabilizers. In other embodiments, soft capsules
contain one or more active compound that is dissolved or suspended
in a suitable liquid. Suitable liquids include, by way of example
only, one or more fatty oil, liquid paraffin, or liquid
polyethylene glycol. In addition, stabilizers are optionally
added.
[0598] In other embodiments, therapeutically effective amounts of
the compound and/or additional therapeutic agent described herein
are formulated for buccal or sublingual administration.
Formulations suitable for buccal or sublingual administration
include, by way of example only, tablets, lozenges, or gels. In
still other embodiments, the compound and/or additional therapeutic
agent described herein are formulated for parental injection,
including formulations suitable for bolus injection or continuous
infusion. In specific embodiments, formulations for injection are
presented in unit dosage form (e.g., in ampoules) or in multi-dose
containers. Preservatives are, optionally, added to the injection
formulations. In still other embodiments, the pharmaceutical
compositions are formulated in a form suitable for parenteral
injection as sterile suspensions, solutions or emulsions in oily or
aqueous vehicles. Parenteral injection formulations optionally
contain formulatory agents such as suspending, stabilizing and/or
dispersing agents. In specific embodiments, pharmaceutical
formulations for parenteral administration include aqueous
solutions of the compound and/or additional therapeutic agent in
water-soluble form. In additional embodiments, suspensions of the
compound and/or additional therapeutic agent are prepared as
appropriate oily injection suspensions. Suitable lipophilic
solvents or vehicles for use in the pharmaceutical compositions
described herein include, by way of example only, fatty oils such
as sesame oil, or synthetic fatty acid esters, such as ethyl oleate
or triglycerides, or liposomes. In certain specific embodiments,
aqueous injection suspensions contain substances which increase the
viscosity of the suspension, such as sodium carboxymethyl
cellulose, sorbitol, or dextran. Optionally, the suspension
contains suitable stabilizers or agents which increase the
solubility of the compound and/or additional therapeutic agent to
allow for the preparation of highly concentrated solutions.
Alternatively, in other embodiments, the active ingredient is in
powder form for constitution with a suitable vehicle, e.g., sterile
pyrogen-free water, before use.
[0599] In still other embodiments, the compound and/or additional
therapeutic agent are administered topically. The compound and/or
additional therapeutic agent described herein are formulated into a
variety of topically administrable compositions, such as solutions,
suspensions, lotions, gels, pastes, medicated sticks, balms, creams
or ointments. Such pharmaceutical compositions optionally contain
solubilizers, stabilizers, tonicity enhancing agents, buffers and
preservatives.
[0600] In yet other embodiments, the compound and/or additional
therapeutic agent are formulated for transdermal administration. In
specific embodiments, transdermal formulations employ transdermal
delivery devices and transdermal delivery patches and can be
lipophilic emulsions or buffered, aqueous solutions, dissolved
and/or dispersed in a polymer or an adhesive. In various
embodiments, such patches are constructed for continuous,
pulsatile, or on demand delivery of pharmaceutical agents. In
additional embodiments, the transdermal delivery of the compound
and/or additional therapeutic agent is accomplished by means of
iontophoretic patches and the like. In certain embodiments,
transdermal patches provide controlled delivery of the compound
and/or additional therapeutic agent. In specific embodiments, the
rate of absorption is slowed by using rate-controlling membranes or
by trapping the compound within a polymer matrix or gel. In
alternative embodiments, absorption enhancers are used to increase
absorption. Absorption enhancers or carriers include absorbable
pharmaceutically acceptable solvents that assist passage through
the skin. For example, in one embodiment, transdermal devices are
in the form of a bandage comprising a backing member, a reservoir
containing the compound optionally with carriers, optionally a rate
controlling barrier to deliver the compound to the skin of the host
at a controlled and predetermined rate over a prolonged period of
time, and means to secure the device to the skin.
[0601] In other embodiments, the compound and/or additional
therapeutic agent are formulated for administration by inhalation.
Various forms suitable for administration by inhalation include,
but are not limited to, aerosols, mists or powders. Pharmaceutical
compositions of compound and/or additional therapeutic agent are
conveniently delivered in the form of an aerosol spray presentation
from pressurized packs or a nebulizer, with the use of a suitable
propellant (e.g., dichlorodifluoromethane, trichlorofluoromethane,
dichlorotetrafluoroethane, carbon dioxide or other suitable gas).
In specific embodiments, the dosage unit of a pressurized aerosol
is determined by providing a valve to deliver a metered amount. In
certain embodiments, capsules and cartridges of, such as, by way of
example only, gelatin for use in an inhaler or insufflator is
formulated containing a powder mix of the compound and a suitable
powder base such as lactose or starch.
[0602] In still other embodiments, the compound and/or additional
therapeutic agent are formulated in rectal compositions such as
enemas, rectal gels, rectal foams, rectal aerosols, suppositories,
jelly suppositories, or retention enemas, containing conventional
suppository bases such as cocoa butter or other glycerides, as well
as synthetic polymers such as polyvinylpyrrolidone, PEG, and the
like. In suppository forms of the compositions, a low-melting wax
such as, but not limited to, a mixture of fatty acid glycerides,
optionally in combination with cocoa butter is first melted.
[0603] In certain embodiments, pharmaceutical compositions are
formulated in any conventional manner using one or more
physiologically acceptable carriers comprising excipients and
auxiliaries which facilitate processing of the active compounds
into preparations which can be used pharmaceutically. Proper
formulation is dependent upon the route of administration chosen.
Any pharmaceutically acceptable techniques, carriers, and
excipients are optionally used as suitable. Pharmaceutical
compositions comprising the compound and/or additional therapeutic
agent are manufactured in a conventional manner, such as, by way of
example only, by means of conventional mixing, dissolving,
granulating, dragee-making, levigating, emulsifying, encapsulating,
entrapping or compression processes.
[0604] Pharmaceutical compositions include at least one
pharmaceutically acceptable carrier, diluent or excipient and at
least the compound and/or additional therapeutic agent, described
herein as an active ingredient. The active ingredient is in
free-acid or free-base form, or in a pharmaceutically acceptable
salt form. In addition, the methods and pharmaceutical compositions
described herein include the use of N-oxides, crystalline forms
(also known as polymorphs), as well as active metabolites of these
compounds having the same type of activity. All tautomers of the
compound and/or additional therapeutic agent described herein are
included within the scope of the compounds presented herein.
Additionally, the compound and/or additional therapeutic agent
described herein encompass unsolvated as well as solvated forms
with pharmaceutically acceptable solvents such as water, ethanol,
and the like. The solvated forms of the compound and/or additional
therapeutic agent presented herein are also considered to be
disclosed herein. In addition, the pharmaceutical compositions
optionally include other medicinal or pharmaceutical agents,
carriers, adjuvants, such as preserving, stabilizing, wetting or
emulsifying agents, solution promoters, salts for regulating the
osmotic pressure, buffers, and/or other therapeutically valuable
substances.
[0605] Methods for the preparation of compositions comprising the
compound and/or additional therapeutic agent described herein
include formulating the compound and/or additional therapeutic
agent with one or more inert, pharmaceutically acceptable
excipients or carriers to form a solid, semi-solid or liquid. Solid
compositions include, but are not limited to, powders, tablets,
dispersible granules, capsules, cachets, and suppositories. Liquid
compositions include solutions in which a compound is dissolved,
emulsions comprising a compound, or a solution containing
liposomes, micelles, or nanoparticles comprising a compound as
disclosed herein. Semi-solid compositions include, but are not
limited to, gels, suspensions and creams. The form of the
pharmaceutical compositions described herein include liquid
solutions or suspensions, solid forms suitable for solution or
suspension in a liquid prior to use, or as emulsions. These
compositions also optionally contain minor amounts of nontoxic,
auxiliary substances, such as wetting or emulsifying agents, pH
buffering agents, and so forth.
[0606] In some embodiments, pharmaceutical composition comprising
the compound and/or additional therapeutic agent illustratively
takes the form of a liquid where the agents are present in
solution, in suspension or both. Typically when the composition is
administered as a solution or suspension a first portion of the
agent is present in solution and a second portion of the agent is
present in particulate form, in suspension in a liquid matrix. In
some embodiments, a liquid composition includes a gel formulation.
In other embodiments, the liquid composition is aqueous.
[0607] In certain embodiments, useful aqueous suspensions contain
one or more polymers as suspending agents. Useful polymers include
water-soluble polymers such as cellulosic polymers, e.g.,
hydroxypropyl methylcellulose, and water-insoluble polymers such as
cross-linked carboxyl-containing polymers. Certain pharmaceutical
compositions described herein comprise a mucoadhesive polymer,
selected for example from carboxymethylcellulose, carbomer (acrylic
acid polymer), poly(methylmethacrylate), polyacrylamide,
polycarbophil, acrylic acid/butyl acrylate copolymer, sodium
alginate and dextran.
[0608] Useful pharmaceutical compositions also, optionally, include
solubilizing agents to aid in the solubility of the compound and/or
additional therapeutic agent. The term "solubilizing agent"
generally includes agents that result in formation of a micellar
solution or a true solution of the agent. Certain acceptable
nonionic surfactants, for example polysorbate 80, are useful as
solubilizing agents, as can ophthalmically acceptable glycols,
polyglycols, e.g., polyethylene glycol 400, and glycol ethers.
[0609] Furthermore, useful pharmaceutical compositions optionally
include one or more pH adjusting agents or buffering agents,
including acids such as acetic, boric, citric, lactic, phosphoric
and hydrochloric acids; bases such as sodium hydroxide, sodium
phosphate, sodium borate, sodium citrate, sodium acetate, sodium
lactate and tris-hydroxymethylaminomethane; and buffers such as
citrate/dextrose, sodium bicarbonate and ammonium chloride. Such
acids, bases and buffers are included in an amount required to
maintain pH of the composition in an acceptable range.
[0610] Additionally, useful compositions also, optionally, include
one or more salts in an amount required to bring osmolality of the
composition into an acceptable range. Such salts include those
having sodium, potassium or ammonium cations and chloride, citrate,
ascorbate, borate, phosphate, bicarbonate, sulfate, thiosulfate or
bisulfite anions; suitable salts include sodium chloride, potassium
chloride, sodium thiosulfate, sodium bisulfite and ammonium
sulfate.
[0611] Other useful pharmaceutical compositions optionally include
one or more preservatives to inhibit microbial activity. Suitable
preservatives include mercury-containing substances such as merfen
and thiomersal; stabilized chlorine dioxide; and quaternary
ammonium compounds such as benzalkonium chloride,
cetyltrimethylammonium bromide and cetylpyridinium chloride.
[0612] Still other useful compositions include one or more
surfactants to enhance physical stability or for other purposes.
Suitable nonionic surfactants include polyoxyethylene fatty acid
glycerides and vegetable oils, e.g., polyoxyethylene (60)
hydrogenated castor oil; and polyoxyethylene alkylethers and
alkylphenyl ethers, e.g., octoxynol 10, octoxynol 40.
[0613] Still other useful compositions include one or more
antioxidants to enhance chemical stability where required. Suitable
antioxidants include, by way of example only, ascorbic acid and
sodium metabisulfite.
[0614] In certain embodiments, aqueous suspension compositions are
packaged in single-dose non-re-closeable containers. Alternatively,
multiple-dose re-closable containers are used, in which case it is
typical to include a preservative in the composition.
[0615] In alternative embodiments, other delivery systems for
hydrophobic pharmaceutical compounds are employed. Liposomes and
emulsions are examples of delivery vehicles or carriers useful
herein. In certain embodiments, organic solvents such as
N-methylpyrrolidone are also employed. In additional embodiments,
the compounds described herein are delivered using a
sustained-release system, such as semipermeable matrices of solid
hydrophobic polymers containing the therapeutic agent. Various
sustained-release materials are useful herein. In some embodiments,
sustained-release capsules release the compounds for a few weeks up
to over 100 days. Depending on the chemical nature and the
biological stability of the therapeutic reagent, additional
strategies for protein stabilization are employed.
[0616] In certain embodiments, the formulations described herein
comprise one or more antioxidants, metal chelating agents, thiol
containing compounds and/or other general stabilizing agents.
Examples of such stabilizing agents, include, but are not limited
to: (a) about 0.5% to about 2% w/v glycerol, (b) about 0.1% to
about 1% w/v methionine, (c) about 0.1% to about 2% w/v
monothioglycerol, (d) about 1 mM to about 10 mM EDTA, (e) about
0.01% to about 2% w/v ascorbic acid, (f) 0.003% to about 0.02% w/v
polysorbate 80, (g) 0.001% to about 0.05% w/v. polysorbate 20, (h)
arginine, (i) heparin, (j) dextran sulfate, (k) cyclodextrins, (1)
pentosan polysulfate and other heparinoids, (m) divalent cations
such as magnesium and zinc; or (n) combinations thereof.
[0617] In some embodiments, the concentration of the compound
and/or additional therapeutic agent provided in the pharmaceutical
compositions of the present invention is less than 100%, 90%, 80%,
70%, 60%, 50%, 40%, 30%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%,
12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.4%,
0.3%, 0.2%, 0.1%, 0.09%, 0.08%, 0.07%, 0.06%, 0.05%, 0.04%, 0.03%,
0.02%, 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%,
0.003%, 0.002%, 0.001%, 0.0009%, 0.0008%, 0.0007%, 0.0006%,
0.0005%, 0.0004%, 0.0003%, 0.0002%, or 0.0001% w/w, w/v or v/v.
[0618] In some embodiments, the concentration of the compound
and/or additional therapeutic agent is greater than 90%, 80%, 70%,
60%, 50%, 40%, 30%, 20%, 19.75%, 19.50%, 19.25% 19%, 18.75%,
18.50%, 18.25% 18%, 17.75%, 17.50%, 17.25% 17%, 16.75%, 16.50%,
16.25% 16%, 15.75%, 15.50%, 15.25% 15%, 14.75%, 14.50%, 14.25% 14%,
13.75%, 13.50%, 13.25% 13%, 12.75%, 12.50%, 12.25% 12%, 11.75%,
11.50%, 11.25% 11%, 10.75%, 10.50%, 10.25% 10%, 9.75%, 9.50%, 9.25%
9%, 8.75%, 8.50%, 8.25% 8%, 7.75%, 7.50%, 7.25% 7%, 6.75%, 6.50%,
6.25% 6%, 5.75%, 5.50%, 5.25% 5%, 4.75%, 4.50%, 4.25%, 4%, 3.75%,
3.50%, 3.25%, 3%, 2.75%, 2.50%, 2.25%, 2%, 1.75%, 1.50%, 125%, 1%,
0.5%, 0.4%, 0.3%, 0.2%, 0.1%, 0.09%, 0.08%, 0.07%, 0.06%, 0.05%,
0.04%, 0.03%, 0.02%, 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%,
0.004%, 0.003%, 0.002%, 0.001%, 0.0009%, 0.0008%, 0.0007%, 0.0006%,
0.0005%, 0.0004%, 0.0003%, 0.0002%, or 0.0001% w/w, w/v, or
v/v.
[0619] In some embodiments, the concentration of the compound
and/or additional therapeutic agent is in the range from
approximately 0.0001% to approximately 50%, approximately 0.001% to
approximately 40%, approximately 0.01% to approximately 30%,
approximately 0.02% to approximately 29%, approximately 0.03% to
approximately 28%, approximately 0.04% to approximately 27%,
approximately 0.05% to approximately 26%, approximately 0.06% to
approximately 25%, approximately 0.07% to approximately 24%,
approximately 0.08% to approximately 23%, approximately 0.09% to
approximately 22%, approximately 0.1% to approximately 21%,
approximately 0.2% to approximately 20%, approximately 0.3% to
approximately 19%, approximately 0.4% to approximately 18%,
approximately 0.5% to approximately 17%, approximately 0.6% to
approximately 16%, approximately 0.7% to approximately 15%,
approximately 0.8% to approximately 14%, approximately 0.9% to
approximately 12%, approximately 1% to approximately 10% w/w, w/v
or v/v.
[0620] In some embodiments, the concentration of the compound
and/or additional therapeutic agent is in the range from
approximately 0.001% to approximately 10%, approximately 0.01% to
approximately 5%, approximately 0.02% to approximately 4.5%,
approximately 0.03% to approximately 4%, approximately 0.04% to
approximately 3.5%, approximately 0.05% to approximately 3%,
approximately 0.06% to approximately 2.5%, approximately 0.07% to
approximately 2%, approximately 0.08% to approximately 1.5%,
approximately 0.09% to approximately 1%, approximately 0.1% to
approximately 0.9% w/w, w/v or v/v.
[0621] In some embodiments, the amount of the compound and/or
additional therapeutic agent is equal to or less than 10 g, 9.5 g,
9.0 g, 8.5 g, 8.0 g, 7.5 g, 7.0 g, 6.5 g, 6.0 g, 5.5 g, 5.0 g, 4.5
g, 4.0 g, 3.5 g, 3.0 g, 2.5 g, 2.0 g, 1.5 g, 1.0 g, 0.95 g, 0.9 g,
0.85 g, 0.8 g, 0.75 g, 0.7 g, 0.65 g, 0.6 g, 0.55 g, 0.5 g, 0.45 g,
0.4 g, 0.35 g, 0.3 g, 0.25 g, 0.2 g, 0.15 g, 0.1 g, 0.09 g, 0.08 g,
0.07 g, 0.06 g, 0.05 g, 0.04 g, 0.03 g, 0.02 g, 0.01 g, 0.009 g,
0.008 g, 0.007 g, 0.006 g, 0.005 g, 0.004 g, 0.003 g, 0.002 g,
0.001 g, 0.0009 g, 0.0008 g, 0.0007 g, 0.0006 g, 0.0005 g, 0.0004
g, 0.0003 g, 0.0002 g, or 0.0001 g.
[0622] In some embodiments, the amount of the compound and/or
additional therapeutic agent is more than 0.0001 g, 0.0002 g,
0.0003 g, 0.0004 g, 0.0005 g, 0.0006 g, 0.0007 g, 0.0008 g, 0.0009
g, 0.001 g, 0.0015 g, 0.002 g, 0.0025 g, 0.003 g, 0.0035 g, 0.004
g, 0.0045 g, 0.005 g, 0.0055 g, 0.006 g, 0.0065 g, 0.007 g, 0.0075
g, 0.008 g, 0.0085 g, 0.009 g, 0.0095 g, 0.01 g, 0.015 g, 0.02 g,
0.025 g, 0.03 g, 0.035 g, 0.04 g, 0.045 g, 0.05 g, 0.055 g, 0.06 g,
0.065 g, 0.07 g, 0.075 g, 0.08 g, 0.085 g, 0.09 g, 0.095 g, 0.1 g,
0.15 g, 0.2 g, 0.25 g, 0.3 g, 0.35 g, 0.4 g, 0.45 g, 0.5 g, 0.55 g,
0.6 g, 0.65 g, 0.7 g, 0.75 g, 0.8 g, 0.85 g, 0.9 g, 0.95 g, 1 g,
1.5 g, 2 g, 2.5, 3 g, 3.5, 4 g, 4.5 g, 5 g, 5.5 g, 6 g, 6.5 g, 7 g,
7.5 g, 8 g, 8.5 g, 9 g, 9.5 g, or 10 g.
[0623] In some embodiments, the amount of the compound and/or
additional therapeutic agent is in the range of 0.0001-10 g,
0.0005-9 g, 0.001-8 g, 0.005-7 g, 0.01-6 g, 0.05-5 g, 0.1-4 g,
0.5-4 g, or 1-3 g.
D. Kits/Articles of Manufacture
[0624] For use in the therapeutic applications described herein,
kits and articles of manufacture are also provided. In some
embodiments, a kit comprising a KRAS, HRAS or NRAS G12C mutant
modulating compound, an additional therapeutic agent and directions
for use of the compound and the additional therapeutic agent for
treatment of cancer is provided. The compound and the additional
therapeutic agent can be selected from any of those described
herein.
[0625] In some embodiments, such kits comprise a carrier, package,
or container that is compartmentalized to receive one or more
containers such as vials, tubes, and the like, each of the
container(s) comprising one of the separate elements to be used in
a method described herein. Suitable containers include, for
example, bottles, vials, syringes, and test tubes. The containers
are formed from a variety of materials such as glass or
plastic.
[0626] The articles of manufacture provided herein contain
packaging materials. Packaging materials for use in packaging
pharmaceutical products include those found in, e.g., U.S. Pat.
Nos. 5,323,907, 5,052,558 and 5,033,252. Examples of pharmaceutical
packaging materials include, but are not limited to, blister packs,
bottles, tubes, inhalers, pumps, bags, vials, containers, syringes,
bottles, and any packaging material suitable for a selected
formulation and intended mode of administration and treatment. For
example, the container(s) includes the compound and/or additional
therapeutic agent described herein, optionally in a separate
composition or in a combined composition. The container(s)
optionally have a sterile access port (for example the container is
an intravenous solution bag or a vial having a stopper pierceable
by a hypodermic injection needle). Such kits optionally comprising
the compound and/or additional therapeutic agent with an
identifying description or label or instructions relating to its
use in the methods described herein.
[0627] For example, a kit typically includes one or more additional
containers, each with one or more of various materials (such as
reagents, optionally in concentrated form, and/or devices)
desirable from a commercial and user standpoint for use of a
compound described herein. Non-limiting examples of such materials
include, but not limited to, buffers, diluents, filters, needles,
syringes; carrier, package, container, vial and/or tube labels
listing contents and/or instructions for use, and package inserts
with instructions for use. A set of instructions will also
typically be included. A label is optionally on or associated with
the container. For example, a label is on a container when letters,
numbers or other characters forming the label are attached, molded
or etched into the container itself, a label is associated with a
container when it is present within a receptacle or carrier that
also holds the container, e.g., as a package insert. In addition, a
label is used to indicate that the contents are to be used for a
specific therapeutic application. In addition, the label indicates
directions for use of the contents, such as in the methods
described herein. In certain embodiments, the pharmaceutical
compositions are presented in a pack or dispenser device which
contains one or more unit dosage forms containing the compound
and/or additional therapeutic agent. The pack for example contains
metal or plastic foil, such as a blister pack. Or, the pack or
dispenser device is accompanied by instructions for administration.
Or, the pack or dispenser is accompanied with a notice associated
with the container in form prescribed by a governmental agency
regulating the manufacture, use, or sale of pharmaceuticals, which
notice is reflective of approval by the agency of the form of the
drug for human or veterinary administration. Such notice, for
example, is the labeling approved by the U.S. Food and Drug
Administration for prescription drugs, or the approved product
insert. In some embodiments, compositions containing a compound
provided herein formulated in a compatible pharmaceutical carrier
are prepared, placed in an appropriate container, and labeled for
treatment of an indicated condition.
EXAMPLES
Example 1
Strategy for Identifying Signaling Pathways Maintained or
Hyper-Activated Following Kras G12C Inhibition
[0628] Pathways suggesting the need for KRAS G12C targeting
compounds in combination with other cancer therapeutics were
assessed as follows. Western blots were used to analyze downstream
KRAS signaling nodes (AKT, ERK, S6, EGFR/HER2, SRC, MET) and a
transcription factor (STAT3). An assessment following treatment of
mutant cells (H358, Calu-1, MiaPaca2, NCI-H23, SW1463, H1792) with
an exemplary KRAS G12C inhibitor (compound I-272) for 48 hours
showed sustained or induced pathways suggesting targeting with RTK,
PI3K, mTOR, SRC, and JAK/STAT inhibitors (FIG. 1). Sustained or
induced pathways were assessed by comparing untreated cell lines
with those treated by compound I-272.
[0629] Specifically, H358 cell lines showed EGFR and HER2
induction, suggesting additional targeting with RTK inhibitors.
Calu-1 and MiaPaca2 cell lines showed refractory PI3K and STAT3,
both with induction, high SRC levels and c-MET induction suggesting
additional targeting with RTK, PI3K, SRC or JAK/STAT inhibitors.
NCI-H23, SW1463, and H1792 cell lines showed refractory PI3K with
induction, refractory ERK, S6 and STAT3, high SRC levels, c-MET
induction, EGFR induction and other sustained RTKs suggesting
additional targeting with RTK, PI3K, SRC, mTOR, or JAK/STAT.
Example 2
Exemplary Kras G12C Inhibitor in Combination with One of RTK, PI3K,
mTOR, SRC or JAK/STAT Inhibitors
[0630] Cancer therapeutics to target specific pathways induced or
sustained by treatment of KRAS G12C cells with an exemplary KRAS
G12C inhibitor (compound I-272) were assessed and demonstrated as
follows. Therapeutics that are RTK, PI3K, mTOR, SRC or JAK/STAT
inhibitors were selected based on data obtained from testing
described in EXAMPLE 1. Cancer therapeutics were selected based on
their potential to have a synergistic effect on targeted cell
lines. Their synergistic effect was assessed and demonstrated as
follows. Comprehensive growth inhibition combination tests were
performed on mutant cell lines (H358, H1792, Calu-1, SW1463,
SW1573, MiaPaca2, NCI-H23) or control cell line (A549) were tested
with an exemplary G12C inhibitor (compound I-272, dosing range of
0.063 .mu.M-2.0M) alone or with compound I-272 (dosing range of
0.063 .mu.M-2.0 .mu.M) in combination with one of erlotinib (EGFR
inhibitor, dose range of 0.16 .mu.M-5.0 .mu.M), GDC0941 (PI3K
inhibitor, dose range of 0.16 .mu.M-5.0 .mu.M), Dasatinib (SRC
inhibitor, dose range 9.3 nM-300 nM), momelotinib (JAK inhibitor,
dose range of 0.16 .mu.M-5.0 .mu.M), or trametinib (MEK inhibitor,
dose range of 1.5 nM-50 nM). Data generated from 3 day
proliferation assays were assessed by luminescence (n=3) and
percentage of growth inhibition was plotted to create an index of
growth inhibition. The growth inhibition index was then color coded
to show areas where combinations tended to produce increased
percentage growth inhibition. That data set was then converted
using the BLISS synergy index to show synergistic combination
treatments.
[0631] Caspase activity in multiple KRAS G12C mutant cells lines
(H358, H2122, H1792, Calu-1, SW1453, SW1573, MiaPaca2, NCI-H23) or
control cell line (A549) was tested with an exemplary G12C
inhibitor (compound I-272, dose range of 0.063 .mu.M-2.0 .mu.M)
alone or compound I-272 (dose range of 0.063 .mu.M-2.0 .mu.M) in
combination with one of erlotinib (EGFR inhibitor, dose range of
0.16 .mu.M-5.0 .mu.M), GDC0941 (PI3K inhibitor, dose range of 0.16
.mu.M-5.0 .mu.M), Dasatinib (SRC inhibitor, dose range 9.3 nM-300
nM), momelotinib (JAK inhibitor, dose range of 0.16 .mu.M-5.0
.mu.M), or trametinib (MEK inhibitor, dose range of 1.5 nM-50 nM).
Caspase activity was measured using a standard caspase activity
luminescence assay (Capase-Glo, Promega) at 6, 24, and 48 hours
(n>7).
[0632] The maximal caspase activity achieved between 6-48 hours was
reported for each cell line and plotted. The plot was then color
coded to show the combinations that had a synergistic effect. The
color coding produced an apoptosis induction index that was used to
assess the combination of therapies. In the H358 cell line,
combinations that induced superior apoptosis compared to treatment
with a single compound or agent include compound I-272 in
combination with one of an EGFR inhibitor, a PI3K inhibitor, a
JAK/TBK1 inhibitor, or an IGF1R inhibitor. In the H2122 cell line,
combinations that induced superior apoptosis compared to treatment
with a single compound or agent include compound I-272 in
combination with one of an EGFR inhibitor, a MEK inhibitor, a PI3K
inhibitor, or an IGF1R inhibitor. In the H1792 cell line,
combinations that induced superior apoptosis compared to treatment
with a single compound or agent include compound I-272 in
combination with one of a JAK/TBK1 inhibitor, a SRC inhibitor, an
EGFR inhibitor, or an IGF1R inhibitor. In the Calu-1 cell line,
combinations that induced superior apoptosis compared to treatment
with a single compound or agent include compound I-272 in
combination with an SRC inhibitor. In the SW1453 cell line,
combinations that induced superior apoptosis compared to treatment
with a single compound or agent include compound I-272 in
combination with one of an EGFR inhibitor or a MEK inhibitor. In
the SW1573 cell line, combinations that induced superior apoptosis
compared to treatment with a single compound or agent include
compound I-272 in combination with an SRC inhibitor. In the
MiaPaca2 cell line, combinations that induced superior apoptosis
compared to treatment with a single compound or agent include
compound I-272 in combination with one of a PI3K inhibitor, a
JAK/TBK1 inhibitor, or an SRC inhibitor.
[0633] No levels of apoptotic induction were observed for any
combination of a KRAS G12C inhibitor with any one chemotherapeutic
agent tested in A549. While not wishing to be bound by theory, it
is believed that this data indicates that the synergistic effects
in H358, H2122, H1792, Calu-1, SW1453, SW1573, MiaPaca2, and
NCI-H23 cell lines are mediated by KRAS G12C specific
inhibition.
[0634] Using the BLISS synergy index data and the apoptosis
induction index data in concert, the best combinations of an
exemplary G12C inhibitor (compound I-272) and one other
chemotherapeutic agent was selected for each cell line. In the H358
cell line, the combination of I-272 with one of erlotinib (EGFR
inhibitor) or GDC0941 (PI3K inhibitor) was selected. In the H1792
cell line, the combination of I-272 with one of dasatinib (SRC
inhibitor) or momelotinib (JAK inhibitor) was selected. In cell
line Calu-1, the combination of I-272 with dasatinib (SRC
inhibitor) was selected. In cell line SW1463, the combination of
I-272 and one of erlotinib (EGFR inhibitor) or GDC0941 (PI3K
inhibitor) was selected. In the cell line SW1573, the combination
of I-272 and one of GDC0941 (PI3K inhibitor) or dasatinib (SRC
inhibitor) was selected. In the cell line MiaPaca2, the combination
of I-272 and one of GDC0941 (PI3K inhibitor) or momelotinib (JAK
inhibitor) was selected. In the cell line NCI-H23, the combination
of I-272 and one of dasatinib (SRC inhibitor) or momelotinib (JAK
inhibitor) was selected. The most frequently synergistic pairs
observed across the cell lines showed combinations of I-272 and one
of a PI3K inhibitor, a SRC inhibitor, or an EGFR inhibitor.
Example 3
Exemplary Kras G12C Inhibitor Used in Combination with One of an
EGFR Inhibitor or PI3K Inhibitor for Synergistic Induction of
Apoptosis
[0635] The effectiveness of KRAS G12C targeting compounds in
combination with other cancer therapeutics was assessed and
demonstrated as follows. Western blots were used to analyze
downstream KRAS signaling nodes (AKT, ERK, RSK, S6) and a marker of
apoptosis (cleaved PARP). Treatment of H358 cells (KRAS G12C) with
exemplary KRAS G12C inhibitors (compounds II-64, I-153 and I-158)
alone for 24 hours causes clear and nearly complete inhibition of
p-ERK, p-RSK, and p-S6, with partial inhibition of p-AKT (FIG. 2A).
However, minimal cleaved PARP is seen, suggesting low levels of
apoptosis (FIG. 2A, lanes 2-4 compared to lane 1). Likewise,
treatment with erlotinib (EGFR inhibitor, lane 5) or GDC0941 (class
I PI3K inhibitor, FIG. 2A, lane 9) alone does not induce robust
apoptosis based on cleaved PARP levels (FIG. 2A, lanes 5 and 9
compared to lane 1). Combination treatment with a KRAS G12C
inhibitor and either erlotinib (FIG. 2A, lanes 6-8) or GDC0941
(FIG. 2A, lanes 9-11) leads to greatly enhanced apoptosis based on
cleaved PARP levels.
[0636] As a control, a non-G12C cell line (i.e., A549) was
subjected to the same single agent and combination treatments (FIG.
2B). The KRAS G12C inhibitors show no single agent or
additive/synergistic effects in this line. While not wishing to be
bound by theory, it is believed that this data indicates that the
synergistic effects in H358 cells are mediated by KRAS G12C
specific inhibition.
[0637] The ability of KRAS G12C targeting compounds in combination
with other cancer therapeutics in inducing apoptosis was assessed
and demonstrated as follows. Caspase activity in multiple KRAS G12C
mutant cells lines (H358, H2122, H2030, H1792, Calu-1, MiaPaca2,
and NCI-H23) or control cell lines (A549 G12S, A375 KRAS WT,
NCI-H411 KRAS G12V, and the HCT115 G13D) was tested with compound
II-64 alone or compound II-64 in combination with one of erlotinib,
afatinib, PI3K (GDC0941), docetaxel, SN38 (active metabolite of
irinotecan), Taxol, IGFIRi (NVP-AEW541), or MEKi (trametinib).
Caspase activity was measured using a standard caspase activity
luminescence assay (Capase-Glo, Promega). Taxol (paclitaxel) was
used as a positive control.
[0638] Three out of seven tested KRAS G12C mutant cell lines
exhibited synergistic induction of apoptosis when a KRAS G12C
inhibitor was combined with an EGFR inhibitor (erlotinib, FIG. 3A).
Four out of seven tested KRAS G12C mutant cell lines exhibit
synergistic induction of apoptosis when a KRAS G12C inhibitor is
combined with a PI3K inhibitor (GDC0941, FIG. 3C). No effect of
KRAS G12C inhibitor is observed in cell line without the KRAS G12C
mutation (FIGS. 3B, 3D). Compound II-64, erlotinib, or GDC0941
alone did not induce significant caspase activity.
[0639] The ability of KRAS G12C targeting compounds in combination
with other cancer therapeutics in inhibiting Ras-mediated cell
cycle progression and induced apoptosis was assessed and
demonstrated as follows. Flow cytometry was used to evaluate KRAS
G12C inhibitor combination treatments. Treatment of H358 cells with
compound II-64, erlotinib, or GDC0941 as single agents leads to G1
arrest with low to modest induction of apoptosis (FIG. 4A,
sub-diploid cell population 8.5-17.8%). Combination treatments
dramatically increase the fraction of apoptotic cells (sub diploid
cell population 40-65%). Similar results are observed for
additional KRAS G12C cell lines (H1792, H2122, SW1573; FIG.
4B).
[0640] Next, flow cytometry was used to evaluate a KRAS G12C
inhibitor in combination with the chemotherapeutic agents
paclitaxel or docetaxel. Synergistic increases in apoptotic
(sub-diploid) H358 cells were observed when compound II-64 was
combined with paclitaxel (FIG. 5), docetaxel (FIG. 6), and SN38
(active form of irinotecan, FIG. 7).
[0641] The ability KRAS G12C targeting compounds in combination
with other cancer therapeutics was assessed and demonstrated as
follows. Calu-1 cells are generally resistant to single agent KRAS
G12C inhibitor as well as combinations with the targeted agents
tested in previous studies disclosed herein (EGFRi, MEKi, PI3Ki,
IGF1Ri; FIG. 7B). Evaluation of phospho-tyrosine levels on a panel
of tyrosine kinases revealed relatively high levels of SRC
phosphorylation in Calu-1 cells (FIG. 8A). Treatment of Calu-1
cells with a KRAS G12C inhibitor (compound I-272) and a SRC
inhibitor (Dasatinib) lead to high levels of apoptosis induction
(FIG. 8B).
Example 4
Exemplary KRAS G12C Inhibitor in Combination with an EGFR, MEK or
Class I PI3K INHIBITOR FOR SYNERGISTIC INDUCTION OF APOPTOSIS
[0642] The effectiveness of KRAS G12C targeting compounds in
combination with other cancer therapeutics was assessed and
demonstrated as follows. Western blots were used to analyze
downstream KRAS signaling nodes (AKT, ERK, RSK, S6) and a marker of
apoptosis (cleaved PARP). Treatment of H358 cells (KRAS G12C) with
an exemplary KRAS G12C inhibitor (compound I-74) alone at a
concentration of 15 .mu.M for 24 hours causes clear and nearly
complete inhibition of p-ERK, p-RSK, and p-S6 (FIG. 9, left panel).
However, minimal cleaved PARP is seen, suggesting low levels of
apoptosis (FIG. 9, left panel, lane 2 compared to lane 9).
Likewise, treatment with erlotinib (EGFR inhibitor, 5 .mu.M, FIG.
9, left panel, lane 3), PD0325901 (MEK inhibitor, 100 nM, FIG. 9,
left panel, lane 5), GDC0941 (class I PI3K inhibitor, 1 .mu.M, FIG.
9, left panel, lane 7) alone does not induce robust apoptosis based
on cleaved PARP levels (FIG. 9, left panel, lanes 3, 5, and 7
compared to lane 9). Combination treatment of H358 cells with
compound I-74, at a concentration of 15 .mu.M, and erlotinib (EGFR
inhibitor, 5 .mu.M, FIG. 9, left panel, lane 4) leads to greatly
enhanced apoptosis based on cleaved PARP levels. An increase in
apoptosis based on cleaved PARP levels is also observed when
compound I-74 (15 .mu.M) is used in combination with PD0325901 (MEK
inhibitor, 100 nM, FIG. 9, left panel, lane 6) or GDC0941 (PI3K
inhibitor, 1 .mu.M, FIG. 9, left panel, lane 8) to treat H358
cells. Taxol (paclitaxel) was used as a positive control.
[0643] As a control, a non-G12C cell line (A549) was subjected to
the same single agent and combination treatments (FIG. 9, right
panel). The KRAS G12C inhibitors show no single agent or
additive/synergistic effects in this line. While not wishing to be
bound by theory, it is believed that this data indicates that the
synergistic effects in H358 cells are mediated by KRAS G12C
specific inhibition.
[0644] Treatment of H358 cells with compound II-74, erlotinib,
PD0325901, or GDC0941 as single agents leads to G1 arrest with low
to modest induction of apoptosis (FIG. 9, left panel), while
combination treatments dramatically increase the fraction of
apoptotic cells.
Example 5
Exemplary KRAS G12C Inhibitor Used in Combination with Pan-PI3K and
Selective PI3Ka for Synergistic Induction of Apoptosis
[0645] Western blots were used to analyze downstream KRAS signaling
nodes (AKT, ERK, RSK, S6) and a marker of apoptosis (cleaved PARP).
Treatment of SW1573 cells with an exemplary KRAS G12C inhibitor
(compound II-64) alone for 24 hours causes inhibition of p-ERK,
p-RSK, and KRAS-GTP (FIG. 10). However, minimal cleaved PARP is
seen, suggesting low levels of apoptosis (FIG. 10, lanes 2-3
compared to lane 1). Likewise, treatment with GDC0941 (class I PI3K
inhibitor, FIG. 10, lane 4) and BYL-719 (selective PI3Ka inhibitor,
FIG. 10, lane 7) alone does not induce robust apoptosis based on
cleaved PARP levels (FIG. 10, lanes 7 compared to lane 1).
Combination treatment with a KRAS G12C inhibitor and either GDC0941
(class I PI3K inhibitor, lanes 5-6) or BYL-719 (selective PI3Ka
inhibitor, FIG. 10, lanes 8-9) leads to greatly enhanced apoptosis
based on cleaved PARP levels. Taxol (paclitaxel) was used as a
positive control. Pan PI3K and selective PI3Ka inhibitors induce
equivalent synergistic apoptosis, measured by cleavage of PARP,
when one is used in combination with a selective KRAS-G12C
inhibitor.
Example 6
Exemplary Kras G12C Inhibitor in Combination of One of an EGFR,
EGFR/HER2, or PI3K Inhibitor for Synergistic Apoptosis and Pathway
Inhibition
[0646] Western blots were used to analyze downstream KRAS signaling
nodes (AKT, ERK, RSK, S6) and a marker of apoptosis (cleaved PARP).
Treatment of H358 cells (KRAS G12C) with an exemplary G12C
inhibitor (compound II-64) alone at 2.5 .mu.M for 24 hours causes
clear and nearly complete inhibition of p-ERK and p-S6, with
partial inhibition of p-AKT (FIG. 11). However, minimal c-caspase 3
is seen, suggesting low levels of apoptosis (FIG. 11, lanes 2
compared to lane 1). Likewise, treatment with erlotinib (EGFR
inhibitor, lane 3), afatinib (MEK inhibitor, FIG. 11, lane 5)
GDC0941 (class I PI3K inhibitor, FIG. 11, lane 7), and selumetinib
(MEK inhibitor, FIG. 11, lane 9) alone does not induce robust
apoptosis based on c-caspase 3 levels (FIG. 11, lanes 3, 5, 7, and
9 compared to lane 1). However, combination treatment with compound
I-64 and either erlotinib (EGFR inhibitor, FIG. 11, lane 4),
afatinib (MEK inhibitor, FIG. 11, lane 6), GDC0941 (class I PI3K
inhibitor, FIG. 11, lane 8), or selumetinib (MEK inhibitor, FIG.
11, lane 10) leads to greatly enhanced apoptosis based on c-caspase
3 levels. Caspase activity was measured using a standard caspase
activity luminescence assay (Capase-Glo, Promega).
Example 7
Exemplary KRAS G12C Inhibitor Used in Combination with an SRC
Inhibitor for Synergistic Induction of Apoptosis of Calu-1
Cells
[0647] Western blots were used to analyze downstream KRAS signaling
nodes (AKT, ERK, RSK, S6) and a marker of apoptosis (cleaved PARP).
Treatment of Calu-1 cells with an exemplary KRAS G12C inhibitor
(compound I-272) alone for 24 hours at concentrations of 30, 100,
300, and 1000 nM causes clear and, at higher concentrations, nearly
complete inhibition of p-ERK and p-RSK (FIG. 12). However, minimal
cleaved PARP is seen, suggesting low levels of apoptosis (FIG. 12,
lanes 2-5 compared to lane 1). Likewise, treatment with dasatinib
(SRC inhibitor, FIG. 12, lane 6) alone does not induce robust
apoptosis based on cleaved PARP levels (FIG. 12, lane 6 compared to
lane 1).
[0648] In contrast, combination treatment with compound I-272
(dosed at 30, 100, 300, and 1000 nM, FIG. 12, lanes 7-10
respectively) and Dasatinib (SRC inhibitor, dosed at 150 nM, FIG.
12, lanes 7-10) leads to greatly enhanced apoptosis based on
cleaved PARP levels. In addition, the combination treatment shows
clear and nearly complete inhibition of p-SRC, p-AKT, p-ERK, p-RSK,
and p-S6.
Example 8
Exemplary Kras G12C Inhibitor Used in Combination with an SRC
Inhibitor for Synergistic Induction of Apoptosis
[0649] Western blots were used to analyze downstream KRAS signaling
nodes (AKT, ERK, RSK, S6) and a marker of apoptosis (cleaved PARP).
Treatment of mutant cell lines (H358, NCI-H23, SW1463, H1792,
Calu-1, SW1573) or a control cell line (A549) was tested with an
exemplary KRAS G12C inhibitor (compound I-272, 1 .mu.M) alone or in
combination with one of dasatinib ("Das," SRC inhibitor, 100 nM) or
Saracatinib ("Sarc," SRC inhibitor, 2 .mu.M). Treatment of mutant
cell lines with KRAS G12C inhibitor, compound I-272, alone for 24
hours causes clear and, in some cases (H358, SW1463, Calu-1),
nearly complete inhibition of p-ERK. The same treatment also shows
inhibition of p-S6 in H358, SW1463, H1792, and Calu-1 cell lines
(FIG. 13).
[0650] However, minimal cleaved PARP is seen, suggesting low levels
of apoptosis. Likewise, treatment with Dasatinib or Sarcatinib
alone does not induce robust apoptosis based on cleaved PARP
levels. In contrast, combination treatment with a KRAS G12C
inhibitor (compound I-272) and either dasatinib or Sarcatinib leads
to greatly enhanced apoptosis based on cleaved PARP levels.
Additionally, the combination of compound I-272 (1 .mu.M) with
dasatinib fully reduces p-S6 in the Calu-1 and SW1573 cell lines.
As a control, a non-G12C cell line (A549) was subjected to the same
single agent and combination treatments. The KRAS G12C inhibitors
show no single agent or additive/synergistic effects in this line.
While not wishing to be bound by theory, it is believed that this
data indicates that the synergistic effects in mutant cell lines
(i.e., H358, NCI-H23, SW1463, H1792, Calu-1, SW1573) are mediated
by KRAS G12C specific inhibition.
[0651] FIG. 14 provides densitometry data for the gels of FIG.
13.
Example 9
Assessing and Demonstrating the Mechanism of Action for Synergistic
Induction of Apoptosis of an Exemplary KRAS G12C Inhibitor Used in
Combination with a JAK Inhibitor in Multiple Mutant Cell Lines
[0652] Western Blots were Used to Analyze Downstream KRAS Signaling
Nodes (AKT, ERK, S6), a transcription factor (STAT3), and a marker
of apoptosis (cleaved PARP). Gel data is provided in FIG. 15.
Treatment of mutant cell lines (H358, H1792, MiaPaca2) or a control
cell line (A549) was tested with an exemplary KRAS G12C inhibitor
(compound I-272, 0.3 and 1 .mu.M) alone or in combination with
momelotinib (JAK inhibitor, 5 .mu.M). Treatment of mutant cell
lines with KRAS G12C inhibitor, compound I-272, alone for 24 hours
causes some and nearly complete inhibition of p-ERK (in H1792 and
H358 respectfully), some inhibition of p-ANK (in H358), some to
nearly complete inhibition of S6 (in H358) as well as some
induction of apoptosis as observed by the presence of cleaved PARP
(in H358 and MiaPaca2). Treatment with I-272 alone also induces
STAT3 in certain cell lines (slight in H358 and H1792, strong in
MiaPaca2 and A549). The combination of compound I-272 (0.3 and 1
.mu.M) and momelotinib (JAK inhibitor, 5 .mu.M) induces apoptosis
in a broad range of G12C positive cell lines and potentiates
inhibition of p-AKT and p-S6. Specifically, when compound I-272 is
used in combination with momelotinib (JAK inhibitor) PARP is
detected in H358, H1792, and MiaPaca2.
[0653] As a control, a non-G12C cell line (A549) was subjected to
the same single agent and combination treatments. The KRAS G12C
inhibitors show no single agent or additive/synergistic effects in
this line. While not wishing to be bound by theory, it is believed
that this data indicates that the synergistic effects in the H358,
H1792, and MiaPaca2 cell lines are mediated by KRAS G12C specific
inhibition.
Example 10
Assessing and Demonstrating the Mechanism of Action for Synergistic
Induction of Apoptosis of an Exemplary Kras G12C Inhibitor Used in
Combination with a JAK Inhibitor in Mutant Cell Lines
[0654] Western blots were used to analyze downstream KRAS signaling
nodes (AKT, ERK, S6), transcription factors (STAT3,
I.kappa.B.alpha.), protein marker (TBK1) and a marker of apoptosis
(cleaved PARP). Treatment of mutant cell lines (NCI-H23, SW1573)
were tested with an exemplary KRAS G12C inhibitor (compound I-272,
1 .mu.M) alone or in combination with one of ruxolitinib (JAK 1/2
inhibitor, 1 and 5 .mu.M) or momelotinib (JAK1/2, TBK1, IKKe
inhibitor, 1 and 5 .mu.M). Treatment of mutant cell lines with KRAS
G12C inhibitor, compound I-272, alone for 24 hours causes clear and
nearly complete inhibition of p-ERK. The same treatment also shows
inhibition of p-S6 and p-AKT in NCI-H23. However, minimal cleaved
PARP is seen, suggesting low levels apoptosis (FIG. 16). Likewise,
treatment with ruxolitinib (JAK1/2 inhibitor, FIG. 16, lane 2-3) or
momelotinib (JAK1/2, TBK1, IKKe inhibitor, FIG. 16, lane 7-8) alone
does not induce robust apoptosis based on cleaved PARP levels (FIG.
16, lanes 2-3 and 7-8 compared to lane 1). Combination treatment
with either ruxolitinib (JAK1/2, TBK1, IKKe inhibitor, FIG. 16,
lane 5-6) or momelotinib (JAK1/2, TBK1, IKKe inhibitor, FIG. 16,
lane 9-10) leads to greatly enhanced apoptosis based on PARP
levels. Further, there is synergistic p-S6 inhibition in each of
the combinations. Synergistic apoptosis strongly occurred when TBK1
and non-canonical NF.kappa.B signaling is inhibited. This effect
suggests the mechanism of action of apoptosis in these combination
treatments is independent from JAK/STAT.
[0655] The following synthetic examples are provided for exemplary
purposes. Other compounds of structures (I), (II) and (III) were
prepared according to analogous procedures.
Example 11
Synthesis of
1-(4-(7-chloro-6-(2-chlorophenyl)quinazolin-4-yl)piperazin-1-yl)prop-2-en-
-1-one (I-1)
##STR00624##
[0656] Compound I-1 was prepared according to Method A as described
below:
Methyl 2-amino-5-(2-chlorophenyl)-4-chlorobenzoate
[0657] A mixture of methyl 2-amino-5-bromo-4-chlorobenzoate (1.2 g,
4.54 mmol), 2-chlorophenylboronic acid (0.85 g, 5.44 mmol),
Na.sub.2CO.sub.3 (1.44 g, 13.61 mmol), and Pd(PPh.sub.3).sub.4(0.52
g, 0.45 mmol) in 1,4-dioxane (30 mL) and water (6 mL) was stirred
at 75.degree. C. under argon for 16 h. The mixture was allowed to
cool to room temperature (RT), and concentrated in vacuo. The
residue was purified by flash column chromatography on silica gel
(ethyl acetate/petroleum=8:1) to afford the desired product (1.22
g, 91% yield) as a yellow solid.
7-Chloro-6-(2-chlorophenyl)quinazolin-4-ol
[0658] A mixture of methyl
2-amino-5-(2-chlorophenyl)-4-chlorobenzoate (342 mg, 1.16 mmol),
CH(OMe).sub.3 (306 mg, 2.89 mmol), and NH.sub.4OAc (223 mg, 2.89
mmol) in MeOH (1 mL) in a sealed tube was stirred at 130.degree. C.
for 4.5 h. The mixture was allowed to cool to RT, and concentrated
in vacuo. The residue was purified by flash column chromatography
on silica gel eluting with DCM and MeOH (40:1) to yield the desired
product (277 mg, 82% yield) as a white solid. ESI-MS m/z: 289.2
[M-H].sup.-.
4,7-Dichloro-6-(2-chlorophenyl)quinazoline
[0659] A mixture of 7-chloro-6-(2-chlorophenyl)quinazolin-4-ol (277
mg, 0.95 mmol), PCl.sub.5 (397 mg, 1.90 mmol) and POCl.sub.3 (16
mL) was stirred at reflux for 20 h. The mixture was allowed to cool
to RT, and then concentrated in vacuo to yield the crude product
(1.19 g) as dark oil which was used directly in next step without
further purification.
tert-Butyl-4-(7-chloro-6-(2-chlorophenyl)quinazolin-4-yl)piperazine-1-carb-
oxylate
[0660] The above obtained crude
4,7-dichloro-6-(2-chlorophenyl)quinazoline (1.19 g) was added to
the mixture of tert-butyl piperazine-1-carboxylate (5 g, 26.9 mmol)
and Et.sub.3N (7.76 g, 76.8 mmol) in DCM (200 mL) at 0.degree. C.
and the resulting mixture was stirred at the same temperature for 1
h. The mixture was poured into water (500 mL) and brine (100 mL),
and then dichloromethane (DCM) (200 mL) was added. The mixture was
filtered through filter paper. The organic layer was separated,
dried over Na.sub.2SO.sub.4 and concentrated in vacuo. The residue
was purified by flash column chromatography on silica gel eluting
with DCM and MeOH (30:1) to yield the desired product (184 mg, 42%
yield, 2 steps) as a light yellow oil. ESI-MS m/z: 459.3
[M+H].sup.+.
1-(4-(7-Chloro-6-(2-chlorophenyl)quinazolin-4-yl)piperazin-1-yl)prop-2-en--
1-one
[0661] A mixture of
tertbutyl-4-(7-chloro-6-(2-chlorophenyl)quinazolin-4-yl)piperazine-1-carb-
oxylate (184 mg, 0.40 mmol) and HCl in MeOH (20 mL) was stirred at
RT for 1 h. The mixture was concentrated in vacuo to yield the
crude product (176 mg) as a yellow solid which was used directly in
next step without further purification.
1-(4-(7-Chloro-6-(2-chlorophenyl)quinazolin-4-yl)piperazin-1-yl)prop-2-en--
1-one (1)
[0662] The above obtained crude
1-(4-(7-chloro-6-(2-chlorophenyl)quinazolin-4-yl)piperazin-1-yl)prop-2-en-
-1-one (17 6 mg) was dissolved in Et.sub.3N (450 mg, 4.45 mmol) and
DCM (30 mL) and cooled to 0.degree. C., acryloyl chloride (44 mg,
0.49 mmol) in DCM (50 mL) was added to the mixture. The resulting
mixture was allowed to warm to RT and stirred at RT for 1.5 h. The
reaction mixture was quenched with saturated NaHCO.sub.3 aqueous
solution, and then extracted with ethyl acetate. The organic layer
was washed with saturated NaHCO.sub.3 solution and brine, dried
over Na.sub.2SO.sub.4 and concentrated in vacuo.
[0663] The residue was purified by flash column chromatography on
silica gel eluting with DCM and MeOH (30:1) to yield the desired
product (82 mg, 50% yield, 2 steps) as a yellow solid. .sup.1H NMR
(400 MHz, DMSO-d6) .delta.: 8.75 (s, 1H), 8.03 (s, 1H), 7.96 (s,
1H), 7.62-7.49 (m, 4H), 6.81 (dd, J=10.4, 16.4 Hz, 1H), 6.15 (dd,
J=16.4, 2.4 Hz, 1H), 5.71 (dd, J=10.4, 2.0 Hz, 1H), 3.87-3.72 (m,
8H). ESI-MS m/z: 413.2 [M+H].sup.+.
Example 12
Synthesis of
1-(4-(7-chloro-6-phenylquinazolin-4-yl)piperazin-1-yl)prop-2-en-1-one
(I-18)
##STR00625##
[0664] Compound I-18 was prepared according to Method B as
described below:
6-Bromo-7-chloroquinazolin-4-ol
[0665] A mixture of methyl 2-amino-5-bromo-4-chlorobenzoate (1 g,
3.95 mmol) and NH.sub.2CHO (20 mL) was stirred at 200.degree. C.
for 3 h. The mixture was allowed to cool to RT and quenched with
water. The solid precipitate was collected by filtration and dried
in vacuo to yield the desired product (669 mg, 66% yield) as a
brown solid.
6-Bromo-4,7-dichloroquinazoline
[0666] A mixture of 6-bromo-7-chloroquinazolin-4-ol (669 mg, 2.59
mmol), PCl.sub.5 (1.6 g, 7.78 mmol) and POCl.sub.3 (15 mL) was
stirred at reflux for 16 h. The mixture was allowed to cool to RT
and then concentrated in vacuo to yield the desired product as dark
oil which was used directly in next step without further
purification.
tert-Butyl
4-(6-bromo-7-chloroquinazolin-4-yl)piperazine-1-carboxylate
[0667] The above obtained crude 6-bromo-4,7-dichloroquinazoline was
added to the mixture of tert-butyl piperazine-1-carboxylate (4.82
g, 25.9 mmol) and Et.sub.3N (2.62 g, 25.9 mmol) in DCM (70 mL). The
resulting mixture was stirred at RT for 2 h and then was quenched
with saturated NaHCO.sub.3 aqueous solution. The mixture was
extracted with DCM, washed with saturated NaHCO.sub.3 aqueous
solution and brine, dried over Na.sub.2SO.sub.4 and concentrated in
vacuo. The residue was purified by flash column chromatography on
silica gel eluting with ethyl acetate and petroleum ether (4:1) to
yield the desired product (631 mg, 57% yield, 2 steps) as a yellow
solid. ESI-MS m/z: 429.3 [M+H].sup.+.
tert-Butyl
4-(7-chloro-6-phenylquinazolin-4-yl)piperazine-1-carboxylate
[0668] A mixture of tert-butyl
4-(6-bromo-7-chloroquinazolin-4-yl)piperazine-1-carboxylate (200
mg, 0.47 mmol), phenylboronic acid (115 mg, 0.94 mmol),
Na.sub.2CO.sub.3 solution (2.0 .mu.M, 0.71 mL, 1.41 mmol),
Pd(PPh.sub.3).sub.4(109 g, 0.094 mmol) in 1,4-dioxane (10 mL) was
stirred at reflux under argon for 16 h. The mixture was allowed to
cool to RT, diluted with ethyl acetate, and then washed with
H.sub.2O and brine. The organic layer was dried over
Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was
purified by flash column chromatography on silica gel eluting with
ethyl acetate and petroleum ether (1:4) to yield the desired
product (120 mg, 60% yield) as a yellow oil. ESI-MS m/z: 425.4
[M+H].sup.+.
1-(4-(7-Chloro-6-phenylquinazolin-4-yl)piperazin-1-yl)prop-2-en-1-one
[0669] The title compound was prepared from tert-butyl
4-(7-chloro-6-phenylquinazolin-4-yl)piperazine-1-carboxylate in two
steps following the procedure described in Example 2. .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta.: 8.74 (s, 1H), 8.15 (s, 1H), 7.83 (s,
1H), 7.50-7.45 (m, 5H), 6.58 (dd, J=16.8, 10.4 Hz, 1H), 6.36 (dd,
J=16.4, 1.6 Hz, 1H), 5.77 (dd, J=10.4, 2.0 Hz, 1H), 3.92-3.81 (m,
8H). ESI-MS m/z: 379.3 [M+H].sup.+.
Example 13
Synthesis of
1-(4-(6-chloro-5-(2-chlorophenyl)-1h-indazol-3-ylamino)piperidin-1-yl)pro-
p-2-en-1-one (I-31)
##STR00626##
[0670] Compound I-31 was prepared according to Method C as
described below:
4-Methyl-N'-(2',4,6-trichlorobiphenylcarbonyl)benzenesulfonohydrazide
[0671] To a stirred solution of 2',4,6-trichlorobiphenyl-3-carbonyl
chloride (5.5 g) in toluene at RT, NH.sub.2NHTs (3.8 g, 20.3 mmol)
was added and the resulting mixture was stirred at 75.degree. C.
overnight. The mixture was allowed to cool to RT. The solid was
collected by filtration and dried in vacuo to afford the desired
product (6 g, 75% yield) as a white solid.
2',4,6-Trichloro-N'-tosylbiphenyl-3-carbohydrazonoyl chloride
[0672] A solution of
4-methyl-N'-(2',4,6-trichlorobiphenylcarbonyl)benzenesulfonohydrazide
(2.3 g, 4.5 mmol) in SOCl.sub.2 (5.8 g, 45 mmol) was stirred at
75.degree. C. for 4 h. The mixture was allowed to cool to RT, and
then petroleum ether was added. The resulting mixture was stirred
at 0.degree. C. for 1 h. The precipitate was collected by
filtration and dried in vacuo to afford the desired product (1.6 g,
67% yield) as a white solid.
tert-Butyl
4-((6-chloro-5-(2-chlorophenyl)-1-tosyl-1H-indazol-3-yl)(4-meth-
oxybenzyl)amino) piperidine-1-carboxylate
[0673] To a stirred solution of
2',4,6-trichloro-N'-tosylbiphenyl-3-carbohydrazonoyl chloride (1.6
g, 3.4 mmol) in 100 mL of NMP at RT, tert-butyl
4-(4-methoxybenzylamino)piperidine-1-carboxylate (1.1 g, 3.4 mmol)
was added followed by K.sub.2CO.sub.3 (1.4 g, 10.2 mmol). The
reaction mixture was stirred at 40.degree. C. overnight. The
mixture was allowed to cool to RT, and partitioned between water
and ethyl acetate. The organic layer was dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The residue
was purified by flash column chromatography on silica gel (1-20%
ethyl acetate/petroleum ether) to afford the desired product (550
mg, 23% yield) as a white solid.
tert-Butyl
4-((6-chloro-5-(2-chlorophenyl)-1H-indazol-3-yl)(4-methoxybenzy-
l)amino) piperidine-1-carboxylate
[0674] To a stirred solution of tert-butyl
4-((6-chloro-5-(2-chlorophenyl)-1-tosyl-1H-indazol-3-yl)(4-methoxybenzyl)-
amino) piperidine-1-carboxylate (550 mg, 0.75 mmol) in THF (20 mL)
and water (5 mL) at RT, NaOH (75 mg, 1.87 mmol) was added, and the
resulting mixture was stirred at reflux overnight. The reaction
mixture was cooled to RT and partitioned between water and ethyl
acetate. The organic layer was dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The residue
was purified by flash column chromatography on silica gel (1-10%
ethyl acetate/petroleum ether) to afford the desired product (100
mg, 23% yield) as a white solid. ESI-MS m/z: 581.5 [M+H].sup.+.
6-Chloro-5-(2-chlorophenyl)-N-(piperidin-4-yl)-1H-indazol-3-amine
[0675] A solution of tert-butyl
4-((6-chloro-5-(2-chlorophenyl)-1H-indazol-3-yl)(4-methoxybenzyl)amino)
piperidine-1-carboxylate (100 mg, 0.17 mmol) in 5 mL of TFA was
stirred at reflux for 2 h. The reaction mixture was allowed to cool
to RT and then partitioned between saturated NaHCO.sub.3 aqueous
solution and ethyl acetate. The organic layer was dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo to
afford the desired product (62 mg) as a yellow solid. The crude
product was used directly in the next step without further
purification.
1-(4-(6-Chloro-5-(2-chlorophenyl)-1H-indazol-3-ylamino)piperidin-1-yl)prop-
-2-en-1-one
[0676] To a stirred solution of acrylic acid (12.4 mg, 0.17 mmol)
in 5 mL of DMF at RT,
6-chloro-5-(2-chlorophenyl)-N-(piperidin-4-yl)-1H-indazol-3-amine
(62 mg, 0.17 mmol), HOBT (30 mg, 0.22 mmol), EDCI (42 mg, 0.22
mmol), and TEA (52 mg, 0.51 mmol) were added sequentially. The
reaction mixture was stirred at RT overnight. The mixture was
partitioned between brine and ethyl acetate. The organic layer was
dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in
vacuo. The residue was purified by prep-HPLC to give the desired
product (2 mg, 3% yield) as a white solid. .sup.1H NMR (300 MHz,
DMSO-d6) .delta.: 11.67 (s, 1H), 7.73 (s, 1H), 7.56-7.58 (m, 1H),
7.41-7.47 (m, 2H), 7.42 (s, 1H), 7.36-7.39 (m, 1H), 6.80-6.87 (m,
1H), 6.07 (dd, J=2.5, 16.7 Hz, 1H), 6.04 (d, J=7.3 Hz, 1H), 5.65
(dd, J=2.4, 10.4 Hz, 1H), 4.23 (d, J=12.3 Hz, 1H), 3.98 (d, J=13.6
Hz, 1H), 3.76-3.80 (m, 1H), 3.26 (t, J=13.0 Hz, 1H), 2.97 (t,
J=10.2 Hz, 1H), 2.06 (m, 2H), 1.38 (m, 2H). ESI-MS m/z: 415.1
[M+H].sup.+.
Example 14
Synthesis of
1-(4-(6-chloro-7-(2-chlorophenyl)isoquinolin-1-yl)piperazin-1-yl)prop-2-e-
n-1-one (I-24)
##STR00627## ##STR00628##
[0677] Compound I-24 was prepared according to Method D as
described below:
N-(3-Bromo-4-chlorobenzyl)-2,2-diethoxyethanamine
[0678] To a solution of 3-bromo-4-chlorobenzaldehyde (10.0 g, 45
mmol) and 2,2-diethoxyethanamine (6.68 g, 50 mmol) in 200 mL of DCM
at RT, 0.5 mL of AcOH was added and the resulting mixture was
stirred at RT for 30 min. To this mixture, NaCNBH.sub.3 (8.1 g, 135
mmol) was added in portions and then stirred at RT overnight. The
reaction mixture was portioned between water and DCM. The organic
layer was washed with water (80 mL.times.2) and brine, dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo to
afford the desired product (11 g, 72% yield) as an oil. The crude
product obtained was used directly in the next step without further
purification.
N-(3-Bromo-4-chlorobenzyl)-2,2-diethoxy-N-tosylethanamine
[0679] To a solution of
N-(3-bromo-4-chlorobenzyl)-2,2-diethoxyethanamine (11 g, 33 mmol)
in 100 mL of DCM, pyridine (10 mL) was added and the resulting
mixture was cooled to 0.degree. C. To this mixture, a solution of
4-methylbenzene-1-sulfonyl chloride (6.8 g, 36 mmol) in 50 mL of
DCM was added dropwise. The reaction mixture was allowed to warm to
RT and stirring was continued until conversion was completed. The
reaction mixture was washed twice with HCl aqueous solution (2 M),
sodium bicarbonate solution and brine. The organic layer was dried
over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in
vacuo. The residue was purified by flash column chromatography on
silica gel (5-20% ethyl acetate/petroleum ether) to afford the
desired product (12.5 g, 78% yield). ESI-MS m/z: 490.2
[M+H].sup.+.
7-Bromo-6-chloroisoquinoline
[0680] AlCl.sub.3 (14.9 g) was suspended in DCM at RT, a solution
of N-(3-bromo-4-chlorobenzyl)-2,2-diethoxy-N-tosylethanamine (11.0
g, 22.5 mmol) in 75 mL of DCM was added and the resulting mixture
was stirred overnight. The mixture was poured into ice water, and
extracted with DCM. The combined organic layer was dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The
residue was purified by flash column chromatography on silica gel
(10-40% ethyl acetate/petroleum ether) to afford the desired
product (5 g, 92.5% yield) as a white solid. ESI-MS m/z: 242
[M+H].sup.+.
7-Bromo-6-chloroisoquinoline2-oxide
[0681] To a solution of 7-bromo-6-chloroisoquinoline (5.5 g, 22.8
mmol) in 100 mL of DCM at RT, was added m-chloroperbenzoic acid
(70%, 5.88 g, 34.2 mmol) and the resulting mixture was stirred at
RT overnight. The precipitate was filtered off and rinsed with DCM.
The filtrate was washed with sodium bicarbonate solution. The
layers were separated and the aqueous layer was extracted with DCM.
The combined organic layer was dried with anhydrous
Na.sub.2SO.sub.4 and concentrated in vacuo to afford the desired
product (4.6 g, 79% yield). The crude product was used directly in
the next step without further purification. ESI-MS m/z: 258.2
[M+H].sup.+.
1-(4-(6-chloro-7-(2-chlorophenyl)isoquinolin-1-yl)piperazin-1-yl)prop-2-en-
-1-one
[0682] The title compound was prepared from
7-Bromo-6-chloroisoquinoline2-oxide in five steps following the
procedure described in Example 2. .sup.1H NMR (400 MHz, DMSO-d6)
.delta.: 8.22-8.21 (m, 2H), 8.00 (s, 1H), 7.65-7.47 (m, 5H), 6.87
(dd, J=16.9, 10.5 Hz, 1H), 6.16 (dd, J=16.7, 1.7 Hz, 1H), 5.72 (dd,
J=10.3, 2.1 Hz, 1H), 3.83 (m, 4H), 3.37 (m, 4H). ESI-MS m/z: 412.2
[M+H].sup.+.
Example 15
Synthesis of
1-(4-(7-chloro-6-(2-chlorophenyl)quinolin-4-yl)piperazin-1-yl)prop-2-en-1-
-one (I-27)
##STR00629##
[0683] Compound I-27 was prepared according to Method E as
described below:
Diethyl 2-((3-chloro-4-iodophenylamino)methylene)malonate
[0684] 3-Chloro-4-iodoaniline (3.0 g, 11.8 mmol) and diethyl
2-(ethoxymethylene)malonate (12.78 g, 59.2 mmol) were mixed in a
100 mL single neck flask, and the resulting mixture was heated to
120.degree. C. and stirred for 2.5 h. The mixture was allowed to
cool to RT and purified by flash column chromatography on silica
gel (10-20% ethyl acetate/petroleum ether) to afford the desired
product (3.93 g) as a white solid. ESI-MS m/z: 422.1
[M-H].sup.-.
Ethyl 7-chloro-4-hydroxy-6-iodoquinoline-3-carboxylate
[0685] (E)-diethyl 2-(((3-chloro-4-iodophenyl)imino)methyl)
malonate (2.0 g, 4.73 mmol) was suspended in 30 mL of Ph.sub.2O.
The mixture was stirred at 250.degree. C. for 4 h. The mixture was
allowed to cool to RT and then 100 mL of petroleum ether was added.
The white solid was collected by filtration and rinsed with
petroleum ether (100 mL) to afford the desired product (1.20 g) as
a white solid.
7-Chloro-4-hydroxy-6-iodoquinoline-3-carboxylic acid
[0686] Ethyl 7-chloro-4-hydroxy-6-iodoquinoline-3-carboxylate (1.2
g, 3.18 mmol) was suspended in 10% NaOH aqueous solution (50 mL).
The mixture was stirred at reflux for 3.5 h. The white solid was
slowly dissolved in NaOH solution. After the mixture turned to a
colorless phase, it was kept heating for additional 1 h. The
mixture was allowed to cool to RT, and the white solid was
separated out. The mixture was acidified with con. HCl to adjust
the pH to 2. The white precipitate was collected by filtration and
rinsed with petroleum ether to afford the desired product (1.13 g)
as a white solid.
7-Chloro-6-iodoquinolin-4-ol
[0687] 7-Chloro-4-hydroxy-6-iodoquinoline-3-carboxylic acid (1.134
g, 3.25 mmol) was suspended in 40 mL of Ph.sub.2O. The mixture was
stirred at 250.degree. C. for 3.5 h. The mixture was allowed to
cool to RT and 100 mL of petroleum ether was added. The solid was
collected by filtration, and rinsed with petroleum ether to afford
the desired product (0.92 g) as a white solid.
4,7-Dichloro-6-iodoquinoline
[0688] 7-Chloro-6-iodoquinolin-4-ol (591 mg, 1.94 mmol) was
dissolved in 40 mL of POCl.sub.3 and the mixture was stirred at
reflux for 3 h. The mixture was allowed to cool to RT and
concentrated in vacuo. The residue was poured into a solution of
Et.sub.3N (2.93 g, 29.03 mmol, 15 eq.) in 40 mL of DCM at 0.degree.
C. The mixture was partitioned between ethyl acetate and brine. The
organic layer was dried and concentrated in vacuo. The residue was
purified by flash column chromatography on silica gel (40% ethyl
acetate/petroleum ether) to afford the desired product (895 mg) as
a solid. ESI-MS m/z: 323.9 [M+H].sup.+.
tert-Butyl
4-(7-chloro-6-iodoquinolin-4-yl)piperazine-1-carboxylate
[0689] 4,7-Dichloro-6-iodoquinoline (200 mg, 0.62 mmol) was mixed
with tert-butyl piperazine-1-carboxylate (172 mg, 0.93 mmol) and
Et.sub.3N (250 mg, 2.47 mmol) in 15 mL DMSO. The resulting mixture
was stirred at 80.degree. C. under argon for 16 h. The mixture was
poured into 250 mL of water and 50 mL of brine, and then extracted
with ethyl acetate. The combined organic layer was washed with
brine, dried over Na.sub.2SO.sub.4, and concentrated in vacuo. The
residue was purified by flash column chromatography on silica gel
(20-30% ethyl acetate/petroleum ether) to afford the desired
product (132 mg). ESI-MS m/z: 374.2 [M+H].sup.+.
tert-Butyl
4-(7-chloro-6-(2-chlorophenyl)quinolin-4-yl)piperazine-1-carbox-
ylate
[0690] tert-Butyl
4-(7-chloro-6-iodoquinolin-4-yl)piperazine-1-carboxylate (130 mg,
0.28 mmol) was mixed with (2-chlorophenyl)boronic acid (109 mg,
0.33 mmol), Pd(PPh.sub.3).sub.4(32 mg, 0.028 mmol) and
Na.sub.2CO.sub.3 (88 mg, 0.83 mmol) in 1,4-dioxane (20 mL) and
water (4 mL). The mixture was stirred at 70.degree. C. under argon
for 4 h. The mixture was allowed to cool to RT and concentrated in
vacuo. The residue was purified by flash column chromatography on
silica gel (30-40% ethyl acetate/petroleum ether) to afford the
desired product (100 mg). ESI-MS m/z: 458.3 [M+H].sup.+.
1-(4-(7-Chloro-6-(2-chlorophenyl)quinolin-4-yl)piperazin-1-yl)prop-2-en-1--
one
[0691] tert-butyl
4-(7-chloro-6-(2-chlorophenyl)quinolin-4-yl)piperazine-1-carboxylate
(100 mg, 0.22 mmol) was dissolved in 20% MeOH--HCl solution (20
mL). The mixture was stirred at RT for 1 h. The mixture was
concentrated in vacuo to yield a yellow solid salt (124 mg). The
yellow salt (124 mg, 0.32 mmol) was dissolved in 30 mL of DCM in
the presence of Et.sub.3N (191 mg, 1.89 mmol). The mixture was
cooled to 0.degree. C. and then a solution of acryloyl chloride (32
mg, 0.35 mmol) in DCM (2 mL) was added dropwise. The mixture was
stirred at 0.degree. C. for 30 min. The mixture was concentrated in
vacuo and the residue was purified by flash column chromatography
on silica gel (50-100% ethyl acetate/petroleum ether) to afford the
desired product (35 mg). .sup.1H NMR (300 MHz, DMSO-d6) .delta.:
8.78-8-79 (m, 1H), 8.17 (s, 1H), 7.96 (s, 1H), 7.65-7.51 (m, 4H),
7.10-7.09 (m, 1H), 6.87 (dd, J=16.4, 10.4 Hz, 1H), 6.15 (d, J=16.4
Hz, 1H), 5.71 (d, J=10.4 Hz, 1H), 3.81 (br s, 4H), 3.22 (br s, 4H).
ESI-MS m/z: 412.2 [M+H].sup.+.
Example 16
Synthesis of
4-(4-acryloylpiperazin-1-yl)-7-chloro-6-(4-chlorophenyl)quinoline-3-carbo-
nitrile (I-42)
##STR00630##
[0692] Compound I-42 was prepared according to Method G as
described below:
3-Chloro-4-(4-chlorophenyl)benzenamine
[0693] A mixture of 3-chloro-4-iodobenzenamine (500 mg, 1.97 mmol),
4-chlorophenylboronic acid (324 mg, 2.07 mmol), Na.sub.2CO.sub.3
(627 mg, 5.92 mmol) and Pd(PPh.sub.3).sub.4(228 mg, 0.20 mmol) in
1,4-dioxane (21 mL) and H.sub.2O (4 mL) was stirred at 80.degree.
C. under argon for 16 h. The mixture was allowed to cool to RT and
concentrated in vacuo. The residue was purified by flash column
chromatography on silica gel (ethyl acetate/petroleum ether=5/1) to
afford the desired product (424 mg, 91% yield) as a yellow
solid.
(E)-Ethyl
3-(3-chloro-4-(4-chlorophenyl)-phenylamino)-2-cyanoacrylate
[0694] A mixture of 3-chloro-4-(4-chlorophenyl)benzenamine (250 mg,
1.05 mmol) and (E)-ethyl 2-cyano-3-ethoxyacrylate (186 mg, 1.10
mmol) was stirred at 100.degree. C. for 2 h and then at 130.degree.
C. for 4 h. The mixture was allowed to cool to RT and concentrated
in vacuo. The residue was triturated with ethyl acetate to afford
the desired product (219 mg, 55% yield) as a white solid. ESI-MS
m/z: 359.1 [M-H].sup.-.
7-chloro-6-(4-chlorophenyl)-4-hydroxyquinoline-3-carbonitrile
[0695] A mixture of
(E)-Ethyl-3-(3-chloro-4-(4-chlorophenyl)-phenylamino)-2-cyanoacrylate
(219 mg, 0.608 mmol) in Ph.sub.2O (8 mL) was stirred at 253.degree.
C. for 4 h. The mixture was cooled to RT and poured into petroleum
ether (20 mL). The precipitate was collected by filtration and
washed with petroleum ether (50 mL.times.2) to yield the desired
product (65 mg, 34% yield) as a brown solid.
4-(4-acryloylpiperazin-1-yl)-7-chloro-6-(4-chlorophenyl)quinoline-3-carbon-
itrile
[0696] The title compound was prepared from
7-chloro-6-(4-chlorophenyl)quinolin-4-ol in four steps according to
the procedure described in Example 2. .sup.1H NMR (400 MHz,
DMSO-d6) .delta.: 8.84 (s, 1H), 8.21 (s, 1H), 8.05 (s, 1H),
7.66-7.59 (m, 4H), 6.88 (dd, J=16.8, 10.4 Hz, 1H), 6.17 (dd,
J=16.8, 2.0 Hz, 1H), 5.74 (dd, J=10.4, 2.0 Hz, 1H), 3.83-3.74 (m,
8H). ESI-MS m/z: 437.2 [M+H].sup.+.
Example 17
Synthesis of
1-(4-(5-(4-chlorophenyl)thieno[2,3-d]pyrimidin-4-yl)piperazin-1-yl)prop-2-
-en-1-one (I-22)
##STR00631##
[0697] Compound I-22 was prepared according to Method H as
described below:
tert-Butyl
4-(5-(4-chlorophenyl)thieno[2,3-d]pyrimidin-4-yl)piperazine-1-c-
arboxylate
[0698] A solution of
4-chloro-5-(4-chlorophenyl)thieno[2,3-d]pyrimidine (180 mg, 0.64
mmol), tert-butyl piperazine-1-carboxylate (119 mg, 0.64 mmol) and
diisopropyl amine in THF (6 mL) was stirred at RT overnight. The
mixture was partitioned between DCM and water. The organic layer
was dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuo
to afford the desired product which was used directly in the next
step without further purification.
5-(4-Chlorophenyl)-4-(piperazin-1-yl)thieno[2,3-d]pyrimidine
hydrochloride
[0699] To a suspension of tert-butyl
4-(5-(4-chlorophenyl)thieno[2,3-d]pyrimidin-4-yl)piperazine-1-carboxylate
obtained from the previous step in 1,4-dioxane (10 mL) and MeOH (5
mL), was added a solution of HCl in 1,4-dioxane (4 .mu.M, 1.0 mL).
The mixture was stirred at RT overnight. The mixture was
concentrated in vacuo and the residue was used directly in the next
step without further purification.
1-(4-(5-(4-Chlorophenyl)thieno[2,3-d]pyrimidin-4-yl)piperazin-1-yl)prop-2--
en-1-one
[0700] To a solution of
5-(4-Chlorophenyl)-4-(piperazin-1-yl)thieno[2,3-d]pyrimidine
hydrochloride obtained above in DCM (10 mL) at 0.degree. C.,
Et.sub.3N (0.2 mL) was added followed by acryloyl chloride. The
resulting mixture was allowed to warm to RT and stirred for 1 h.
The mixture was partitioned between DCM and water. The organic
layer was dried over Na.sub.2SO.sub.4, filtered and concentrated in
vacuo. The residue was purified via Isolera One (silica cartridge,
0-60% ethyl acetate/hexanes) to afford the desired product (27.5
mg). .sup.1H NMR (300 MHz, CDCl.sub.3), .delta.: 8.64 (s, 1H),
7.35-7.48 (m, 4H), 7.30 (s, 1H), 6.42-6.60 (m, 1H), 6.26 (d, J=24
Hz, 1H), 5.69 (d, J=10.5 Hz, 1H), 3.10-3.35 (m, 8H). ESI-MS m/z:
385.0 [M+H].sup.+
Example 18
Synthesis of
1-(4-(8-(2-chlorophenyl)quinazolin-2-yl)piperazin-1-yl)prop-2-en-1-one
(I-35)
##STR00632##
[0701] Compound I-35 was prepared according to Method I as
described below: tert-Butyl
4-(8-bromoquinazolin-2-yl)piperazine-1-carboxylate
[0702] The title compound was prepared from
8-bromo-2-chloroquinazoline according to the procedure described in
step 1 in Example 8.
tert-Butyl
4-(8-(2-chlorophenyl)quinazolin-2-yl)piperazine-1-carboxylate
[0703] A mixture of tert-Butyl
4-(8-bromoquinazolin-2-yl)piperazine-1-carboxylate (250 mg, 0.64
mmol), 2-chlorophenylbronic acid (110 mg, 1.1 mmol) and
Pd(dppf)Cl.sub.2.CH.sub.2Cl.sub.2 (50 mg) in a mixture of
1,4-dioxane (6 mL) and sat. NaHCO.sub.3 solution (3 mL) was stirred
at 100.degree. C. for 1 h. The mixture was allowed to cool to RT,
and partitioned between water and ethyl acetate. The organic layer
was dried over Na.sub.2SO.sub.4, filtered and concentrated in
vacuo. The residue was purified via Isolera One (silica cartridge,
0-60% ethyl acetate/hexanes) to afford the desired product.
1-(4-(8-(2-Chlorophenyl)quinazolin-2-yl)piperazin-1-yl)prop-2-en-1-one
[0704] The title compound was prepared from tert-Butyl
4-(8-(2-chlorophenyl)quinazolin-2-yl)piperazine-1-carboxylate
according to the procedure described in steps 2 and 3 in Example 8.
.sup.1H NMR (300 MHz, CDCl.sub.3) .delta.: 9.07 (s, 1H), 7.74 (dd,
J=8.0, 1.6 Hz, 1H), 7.67 (dd, J=6.8, 1.2 Hz, 1H), 7.46-7.56 (m,
1H), 7.39-7.42 (m, 4H), 6.58 (dd, J=16.8, 10.8 Hz, 1H), 6.32 (dd,
J=16.8, 2.0 Hz, 1H), 5.71 (dd, J=10.6, 1.9 Hz, 1H), 3.8-3.9 (br.,
4H), 3.68-3.78 (br., 2H), 3.55-3.62 (br., 2H). ESI-MS m/z: 379.1
[M+H].sup.+.
Example 19
Synthesis of
1-(4-(5-(2-chlorophenyl)-7h-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)p-
rop-2-en-1-one (I-28)
##STR00633##
[0706] Compound I-28 was prepared according to Method J as
described below:
4-Chloro-7H-pyrrolo[2,3]pyrimidine
[0707] A mixture of 1H-pyrrolo[2,3-d]pyrimidin-4(7H)-one (2.5 g.
18.6 mmol) in 46 mL of POCl.sub.3 was stirred at reflux for 5 h.
The mixture was allowed to cool to RT and then concentrated in
vacuo to remove the excess amount of POCl.sub.3. Ice was added to
the residue and the mixture was stirred at RT for 10 min. The
aqueous layer was extracted with diethyl ether. The organic layer
was dried over MgSO.sub.4, filtered, and concentrated in vacuo to
afford the desired product (1.5 g, 54% yield) as an off-white
solid.
4-Chloro-5-iodo-7H-pyrrolo[2,3]pyrimidine
[0708] 4-Chloro-7H-pyrrolo[2,3-d]pyrimidine (1.8 g 11.9 mmol) and
N-iodosuccinamide (3 g, 13.1 mmol) were mixed in a round bottomed
flask. The flask was dried under high vacuum for 5 h and then
back-filled with argon. To this mixture, dry DMF (100 mL) was added
and the resulting mixture was stirred in the dark for 20 h. The
reaction was quenched with methanol and concentrated in vacuo. The
residue was diluted with 150 mL of DCM and washed with water (200
mL), saturated aqueous sodium sulfite (200 mL), and brine (100 mL).
The organic layer was dried over MgSO.sub.4, filtered, and
concentrated in vacuo. The residue was purified by flash column
chromatography on silica gel (50% ethyl acetate/hexanes) to afford
the desired product (3.1 g, 95% yield) as a white solid. ESI-MS
m/z: 279.5 [M+H].sup.+.
4-Chloro-5-iodo-7benzenesulfonyl-pyrrolo[2,3-d]pyrimidine
[0709] To a solution of 4-chloro-5-iodo-7H-pyrrolo[2,3]pyrimidine
(280 mg, 1 mmol) in DMF (5 mL) at 0.degree. C., NaH (60%, 52 mg,
1.3 mmol) was added and the resulting mixture was stirred at
0.degree. C. for 30 min. To this mixture, benzenesulfonyl chloride
(194 mg, 1.1 mmol) was added. The mixture was then stirred at RT
for 2 h. The mixture was partitioned between ethyl acetate and
water. The organic layer was dried over MgSO.sub.4, filtered, and
concentrated in vacuo. The residue was purified by flash column
chromatography on silica gel to afford the desired product (300 mg,
71.6% yield).
4-Chloro-5-(2-chlorophenyl)-7H-pyrrolo[2,3-d]pyrimidine
[0710] To a solution of
4-chloro-5-iodo-7benzenesulfonyl-pyrrolo[2,3-d]pyrimidine (300 mg,
0.71 mmol) and 2-chlorophenylboronic acid (167 mg, 1.07 mmol) in
1,4-dioxane (15 mL) and water (3 mL), Pd(PPh.sub.3).sub.4(60 mg)
and Na.sub.2CO.sub.3 (227 mg, 2.14 mmol) were added. The mixture
was stirred at 80.degree. C. overnight. The mixture was allowed to
cool to RT and concentrated in vacuo. The residue was purified by
flash column chromatography on silica gel to afford the desired
product (120 mg, 63% yield). ESI-MS m/z: 262.2 [M-H].sup.-.
tert-butyl-4-(5-(2-chlorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-
e-1-carboxylate
[0711] To a solution of
4-chloro-5-(2-chlorophenyl)-7H-pyrrolo[2,3-d]pyrimidine (120 mg,
0.45 mmol) and tert-butyl piperazine-1-carboxylate (254 mg, 1.36
mmol) in 1,4-dioxane (15 mL), DIEA (293 mg, 2.27 mmol) was added.
The mixture was stirred at 100.degree. C. overnight. The mixture
was concentrated in vacuo and the residue was purified by flash
column chromatography on silica gel to afford the desired product
(120 mg, 64% yield).
1-(4-(5-(2-Chlorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)pr-
op-2-en-1-one
[0712] The title compound was prepared from
tert-butyl-4-(5-(2-chlorophenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazi-
ne-1-carboxylate in two steps according to the procedure described
in Example 2. .sup.1H NMR (400 MHz, DMSO-d6) .delta.: 8.5 (s, 1H),
7.5 (m, 1H), 7.4 (m, 3H), 7.3 (s, 2H), 6.5 (m, 1H), 6.3 (m, 1H),
5.7 (m, 1H), 3.4 (m, 8H). ESI-MS m/z: 368.3 [M+H].sup.+.
Example 20
Synthesis of
1-(4-(2-amino-7-chloro-6-(4-chlorophenyl)quinazolin-4-yl)piperazin-1-yl)p-
rop-2-en-1-one (I-39) and
1-(4-(7-chloro-6-(4-chlorophenyl)-2-methoxyquinazolin-4-yl)piperazin-1-yl-
)prop-2-en-1-one (I-43)
##STR00634##
[0714] Compounds 1-39 and 1-43 were prepared according to Method F
as described below:
6-Bromo-7-chloroquinazoline-2,4-diol
[0715] A mixture of methyl 2-amino-5-bromo-4-chlorobenzoate (3.0 g,
11.34 mmol) and urea (1.36 g, 22.68 mmol, 2 eq.) was stirred at
200.degree. C. for 3 h. The mixture was allowed to cool to RT,
triturated with ethyl acetate and dried to afford the desired
product (2.39 g) as a brown solid.
6-Bromo-2,4,7-trichloroquinazoline
[0716] The mixture of 6-bromo-7-chloroquinazoline-2,4-diol (1.1 g,
6.79 mmol) in 30 mL of POCl.sub.3 was stirred at reflux for 2 days.
The mixture was allowed to cool to RT and concentrated in vacuo to
remove POCl.sub.3. The residue was poured into a solution of
Et.sub.3N (13.7 g, 20 eq.) in 30 mL of DCM at 0.degree. C. The
mixture was partitioned between ethyl acetate and brine. The
organic layer was dried over Na.sub.2SO.sub.4, filtered and
concentrated in vacuo. The residue was purified by flash
chromatography on silica gel (5-10% ethyl acetate/petroleum ether)
to afford the desired product (474 mg) as a yellow solid.
tert-Butyl-4-(6-bromo-2,7-dichloroquinazolin-4-yl)piperazine-1-carboxylate
[0717] To a solution of tert-butyl piperazine-1-carboxylate (123
mg, 0.66 mmol) in DMF (10 mL) at RT, DIEA (94 mg, 0.72 mmol) was
added followed by 6-bromo-2,4,7-trichloroquinazoline (206 mg, 0.66
mmol). The resulting mixture was stirred at 50.degree. C. for 40
min. The mixture was allowed to cool to RT and partitioned between
water and ethyl acetate. The organic layer was washed with brine,
dried over Na.sub.2SO.sub.4 and concentrated. The residue was
purified by flash column chromatography on silica gel (5% ethyl
acetate/petroleum ether) to afford the desired product (222 mg) as
a yellow solid. ESI-MS m/z: 463.2 [M+H].sup.+.
tert-Butyl
4-(6-bromo-7-chloro-2-methoxyquinazolin-4-yl)piperazine-1-carbo-
xylate
[0718] To a solution of NaOMe (26 mg, 0.476 mmol) in MeOH (20 mL),
tert-butyl-4-(6-bromo-2,7-dichloroquinazolin-4-yl)piperazine-1-carboxylat-
e (110 mg, 0.238 mmol) was added. The mixture was stirred at
60.degree. C. under argon for 40 min. The mixture was quenched by
water (1.0 mL) and then concentrated in vacuo. The residue was
purified by flash column chromatography on silica gel (10-20% ethyl
acetate/petroleum ether) to afford the desired product (55 mg) as a
yellow solid. ESI-MS m/z: 459.2 [M+H].sup.+.
tert-Butyl-4-(7-chloro-6-(4-chlorophenyl)-2-methoxyquinazolin-4-yl)piperaz-
ine-1-carboxylate
[0719] The mixture of tert-butyl
4-(6-bromo-7-chloro-2-methoxyquinazolin-4-yl)piperazine-1-carboxylate
(85 mg, 0.19 mmol), (4-chlorophenyl)boronic acid (35 mg, 0.22
mmol), Pd(PPh.sub.3).sub.4(22 mg, 0.019 mmol), Na.sub.2CO.sub.3 (60
mg, 0.56 mmol) in dioxane (20 mL) and water (2 mL) was stirred at
80.degree. C. under argon for 16 h. The mixture was allowed to cool
to RT and concentrated in vacuo. The residue was purified by flash
column chromatography on silica gel (10-20% ethyl acetate/petroleum
ether) followed by Prep-TLC to afford the desired product (100 mg)
as a white solid. ESI-MS m/z: 489.4 [M+H].sup.+.
1-(4-(7-Chloro-6-(4-chlorophenyl)-2-methoxyquinazolin-4-yl)piperazin-1-yl)-
prop-2-en-1-one
[0720]
tert-Butyl-4-(7-chloro-6-(4-chlorophenyl)-2-methoxyquinazolin-4-yl)-
piperazine-1-carboxylate (100 mg, 0.20 mmol) was dissolved in 20 mL
of 20% HCl methanol solution. The mixture was stirred at RT for 1 h
and then concentrated in vacuo to yield a yellow solid salt (90
mg).
[0721] The above yellow solid (90 mg, 0.21 mmol) was dissolved in
30 mL of DCM with Et.sub.3N (129 mg, 1.27 mmol). The mixture was
cooled to 0.degree. C. and then added dropwise to a solution of
acryloyl chloride (23 mg, 0.25 mmol) in DCM (2 mL). The resulting
mixture was stirred at 0.degree. C. for 30 min. The mixture was
poured into H.sub.2O (100 mL), sat. NaHCO.sub.3 (50 mL) and brine
(50 mL), and then extracted with ethyl acetate. The organic layer
was washed with brine, dried over Na.sub.2SO.sub.4 and
concentrated. The residue was purified by Prep-TLC followed by
Prep-HPLC to afford the desired product (8 mg) as a white solid.
ESI-MS m/z: 443.2 [M+H].sup.+.
tert-Butyl-4-(2-amino-6-bromo-7-chloroquinazolin-4-yl)piperazine-1-carboxy-
late
[0722] The mixture of tert-butyl
4-(6-bromo-2,7-dichloroquinazolin-4-yl)piperazine-1-carboxylate in
sat. NH.sub.3-EtOH (4 mL) in a sealed tube was stirred at
100.degree. C. for 16 h. The mixture was allowed to cool to RT and
concentrated in vacuo. The residue was purified by flash column
chromatography on silica gel (20-30% ethyl acetate/petroleum ether)
to afford the desired product (70 mg) as a white solid.
tert-Butyl-4-(2-amino-7-chloro-6-(4-chlorophenyl)quinazolin-4-yl)piperazin-
e-1-carboxylate
[0723] The mixture of
tert-butyl-4-(2-amino-6-bromo-7-chloroquinazolin-4-yl)piperazine-1-carbox-
ylate (70 mg, 0.16 mmol), (4-chlorophenyl)boronic acid (29 mg, 0.19
mmol), Pd(PPh.sub.3).sub.4(18 mg, 0.019 mmol), and Na.sub.2CO.sub.3
(50 mg, 0.48 mmol) in dioxane (20 mL) and water (2 mL) was stirred
at 80.degree. C. under argon for 16 h. The mixture was allowed to
cool to RT and then concentrated in vacuo. The residue was purified
by flash column chromatography on silica gel (10-20% ethyl
acetate/petroleum ether) followed by Prep-TLC to afford the desired
product (70 mg) as a red solid. ESI-MS m/z: 474.5[M+H].sup.+.
1-(4-(2-Amino-7-chloro-6-(4-chlorophenyl)quinazolin-4-yl)piperazin-1-yl)pr-
op-2-en-1-one
[0724]
tert-Butyl-4-(2-amino-7-chloro-6-(4-chlorophenyl)quinazolin-4-yl)pi-
perazine-1-carboxylate (70 mg, 0.15 mmol) was dissolved in 20% HCl
methanol solution (20 mL) and the resulting mixture was stirred at
RT for 1 h. The mixture was concentrated to afford the desired
product (70 mg) as a yellow solid salt.
[0725] The mixture of above obtained yellow solid (70 mg, 0.21
mmol), acrylic acid (18 mg, 0.25 mmol), EDCI (73 mg, 0.381 mmol)
and HOBT (52 mg, 0.381 mmol) in 10 mL of DMF at 0.degree. C., a
solution of Et.sub.3N (120 mg, 1.2 mmol) in DCM (2 mL) was added
dropwise. The resulting mixture was stirred at 0.degree. C. for 30
min and at RT for 1.5h. The mixture was poured into water (100 mL),
sat. NaHCO.sub.3 (50 mL) and brine (50 mL), and then extracted with
ethyl acetate. The organic layer was washed with brine, dried over
Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was
purified by flash column chromatography on silica gel to yield the
desired product (5 mg) as a gray solid. ESI-MS m/z: 428.3
[M+H].sup.+.
Example 21
Synthesis of
1-(4-(7-chloro-6-(2-chlorophenyl)quinazolin-4-yl)piperidin-1-yl)prop-2-en-
-1-one (I-36)
##STR00635##
[0726] Compound I-36 was prepared according to Method K as
described below:
1-tert-Butyl 4-methyl
4-(6-bromo-7-chloroquinazolin-4-yl)piperidine-1,4-dicarboxylate
[0727] To a stirred solution of tert-butyl methyl
piperidine-1,4-dicarboxylate (3.3 g, 13.5 mmol) in anhydrous THF
(30 mL) at 0.degree. C. under nitrogen, LiHMDS (15 mL, 15 mmol) was
added and the resulting mixture was stirred at 0.degree. C. for 1
h. To this mixture, a solution of 6-bromo-4,7-dichloroquinazoline
(748 mg, 2.7 mmol) in THF (5 mL) was added and the resulting
mixture was stirred at room temperature for 4 h. The mixture was
quenched with ice-water and partitioned between water and ethyl
acetate. The organic layer was dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The residue
was purified by flash column chromatography on silica gel (1-10%
ethyl acetate/petroleum ether) to afford the desired product (580
mg, 37% yield) as a white solid.
tert-Butyl
4-(6-bromo-7-chloroquinazolin-4-yl)piperidine-1-carboxylate
[0728] To a solution of 1-tert-butyl 4-methyl
4-(6-bromo-7-chloroquinazolin-4-yl)piperidine-1,4-dicarboxylate
(483 mg, 1.2 mmol) in DMSO (10 mL), LiCl (103 mg, 2.4 mmol) and
water (65 mg, 3.6 mmol) were added, and the rusting mixture was
stirred at 110.degree. C. for 16 h. The mixture was allowed to cool
to room temperature and partitioned between water and ethyl
acetate. The organic layer was dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The residue
was purified by flash column chromatography on silica gel (1-20%
ethyl acetate/petroleum ether) to afford the desired product (170
mg, 33% yield) as a white solid.
tert-Butyl
4-(7-chloro-6-(2-chlorophenyl)quinazolin-4-yl)piperidine-1-carb-
oxylate
[0729] A mixture of tert-butyl
4-(6-bromo-7-chloroquinazolin-4-yl)piperidine-1-carboxylate (230
mg, 0.59 mmol), 2-chlorophenylboronic acid (138 mg, 0.88 mmol),
Pd(PPh.sub.3).sub.4(69 mg, 0.06 mmol) and Na.sub.2CO.sub.3 (188 mg,
106 mmol) in 1,4-dioxane (10 mL) under argon was stirred at
100.degree. C. for 16 h. The mixture was allowed to cool to room
temperature, and partitioned between water and ethyl acetate. The
organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered
and concentrated in vacuo. The residue was purified by flash column
chromatography on silica gel (1-20% ethyl acetate/petroleum ether)
to afford the desired product (160 mg, 65% yield) as a white
solid.
1-(4-(7-Chloro-6-(2-chlorophenyl)quinazolin-4-yl)piperidin-1-yl)prop-2-en--
1-one (I-36)
[0730] The title compound was prepared from tert-butyl
4-(7-chloro-6-(2-chlorophenyl)
quinazolin-4-yl)piperidine-1-carboxylate according to the procedure
described in steps 5 and 6 in Example 2. .sup.1H NMR (400 MHz,
DMSO-d6) .delta.: 9.28 (s, 1H), 8.55 (s, 1H), 8.27 (s, 1H), 7.70
(m, 2H), 7.53-7.68 (m, 2H), 6.82-6.88 (m, 1H), 6.10 (dd, J=2.5,
16.8 Hz, 1H), 5.68 (dd, J=2.3, 10.3 Hz, 1H), 4.55 (d, J=12.2 Hz,
1H), 4.09-4.16 (m, 2H), 3.32 (t, J=12.2 Hz, 1H), 2.89 (t, J=12.1
Hz, 1H), 1.72-1.93 (m, 4H). ESI-MS m/z: 410.35 [M-H].sup.-.
Example 22
Synthesis of
7-chloro-6-(4-chlorophenyl)-4-(4-(vinylsulfonyl)piperazin-1-yl)quinazolin-
e (I-45)
##STR00636##
[0731] Compound I-45 was prepared according to the general
procedures of Method A as described below:
tert-Butyl
4-(7-chloro-6-(4-chlorophenyl)quinazolin-4-yl)piperazine-1-carb-
oxylate
[0732] The title compound was prepared from tert-butyl
4-(6-bromo-7-chloroquinazolin-4-yl)piperazine-1-carboxylate and
4-chlorophenylboronic acid according to the procedure described in
step 4 in Example 3.
tert-Butyl4-(7-chloro-6-(4-chlorophenyl)quinazolin-4-yl)piperazine-1-carbo-
xylate
[0733] A solution of
tert-butyl4-(7-chloro-6-(4-chlorophenyl)quinazolin-4-yl)piperazine-1-carb-
oxylate (500 mg, 1.09 mmol) in HCl/MeOH (10 mL, 28.6 mmol) was
stirred at room temperature for 30 min. The mixture was
concentrated in vacuo to afford the crude product.
7-Chloro-6-(4-chlorophenyl)-4-(4-(vinylsulfonyl)piperazin-1-yl)quinazoline
[0734] The above obtained crude product was dissolved with DCM (15
mL) and cooled to 0.degree. C. To this mixture,
2-chloroethanesulfonyl chloride (213.2 mg, 1.31 mmol) and Et.sub.3N
(1.5 mL, 10.9 mmol) were added and the resulting mixture was
stirred at 0.degree. C. for 10 min. The mixture was quenched with
ice-water and partitioned between water and ethyl acetate. The
organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered
and concentrated in vacuo. The residue was purified by prep-HPLC to
afford the desired product (3 mg, 0.6% yield). .sup.1H-NMR (400 M
Hz, CDCl.sub.3) .delta.: 8.78 (s, 1H), 8.08 (s, 1H), 7.75 (s, 1H),
7.49 (d, J=8.4 Hz, 2H), 7.42 (d, J=8.4 Hz, 2H), 6.46 (dd, J=10,
16.8 Hz, 1H), 6.31 (d, J=16.8 Hz, 1H), 6.11 (d, J=9.6 Hz, 1H), 3.91
(t, J=4.8 Hz, 4H), 3.35 (t, J=4.8 Hz, 4H). ESI-MS m/z: 449.25
[M+H].sup.+.
Example 23
Synthesis of
1-(4-(7-chloro-6-(4-chlorophenyl)quinazolin-4-yl)-2-(hydroxymethyl)pipera-
zin-1-yl)prop-2-en-1-one (I-46)
##STR00637##
[0735] Compound I-46 was prepared according to the general
procedures of Method A as described below:
4,7-Dichloro-6-(4-chlorophenyl)quinazoline
[0736] The title compound was prepared from
2-amino-5-bromo-4-chlorobenzoate according to the procedure
described in steps 1, 2 and 3 in Example 2.
tert-Butyl
4-(7-chloro-6-(4-chlorophenyl)quinazolin-4-yl)-2-(hydroxymethyl-
)piperazine-1-carboxylate
[0737] The above obtained crude
4,7-dichloro-6-(4-chlorophenyl)quinazoline (200 mg, 0.464 mmol) was
added to the mixture of tert-butyl
2-(hydroxymethyl)piperazine-1-carboxylate (210 mg, 0.968 mmol) and
DIEA (418 mg, 3.24 mmol) in 1,4-dioxane (20 mL) at room temperature
and the resulting mixture was stirred at 80.degree. C. for 3 h. The
mixture was allowed to cool to room temperature and then
concentrated in vacuo. The residue was purified by flash column
chromatography on silica gel (DCM/MeOH=30:1) to afford the desired
product (110 mg, 35% yield) as a light yellow oil. ESI-MS m/z:
498.9 [M+H].sup.+.
(4-(7-Chloro-6-(4-chlorophenyl)quinazolin-4-yl)piperazin-2-yl)methanol
hydrochloride
[0738] A mixture of
4-(7-chloro-6-(4-chlorophenyl)quinazolin-4-yl)-2-(hydroxymethyl)piperazin-
e-1-carboxylate (110 mg, 0.225 mmol) and HCl in MeOH (10 mL, 28.6
mmol) was stirred at room temperature for 1 h. The mixture was
concentrated in vacuo to afford the crude product (106 mg) as a
yellow solid which was used directly in next step without further
purification.
1-(4-(7-Chloro-6-(4-chlorophenyl)quinazolin-4-yl)-2-(hydroxymethyl)piperaz-
in-1-yl)prop-2-en-1-one
[0739] To a stirred solution of above obtained yellow solid (106
mg, 0.225 mmol) in DMF (5 mL) at room temperature, acrylic acid (19
mg, 0.27 mmol), BOP (149 mg, 0.338 mmol) and DIEA (203 mg, 1.58
mmol) were added and the resulting mixture was stirred at room
temperature for 30 min. The mixture was poured into saturated
aqueous NaHCO.sub.3 solution (50 mL), and then extracted with ethyl
acetate. The organic layer was washed with brine, dried over
Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was
purified by flash column chromatography on silica gel
(DCM/MeOH=20:1) to afford the desired product (20 mg, 20% yield, 2
steps) as a solid. .sup.1H NMR (400 MHz, DMSO-d6) .delta.: 8.7 (s,
1H), 8.2 (d, J=2.8 Hz, 1H), 8.0 (s, 1H), 7.5 (m, 4H), 6.8 (dd,
J=10.4, 16.4 Hz, 1H), 6.1 (d, J=17 Hz, 1H), 5.7 (dd, J=2.4, 10.4
Hz, 1H), 5.0 (m, 1H), 4.3 (m, 2H), 4.2 (m, 2H), 3.6 (m, 3H), 2.5
(s, 2H). ESI-MS m/z: 443.30 [M+H].sup.+.
Example 24
Synthesis of
1-acryloyl-4-(7-chloro-6-(4-chlorophenyl)quinazolin-4-yl)piperazine-2-car-
bonitrile (I-47)
##STR00638##
[0740] Compound I-47 was prepared according to the general
procedures of Method A as described below:
4-(7-Chloro-6-(4-chlorophenyl)quinazolin-4-yl)piperazine-2-carboxamide
[0741] The crude 4,7-dichloro-6-(4-chlorophenyl)quinazoline (310
mg, 1 mmol) was added to the mixture of piperazine-2-carboxamide
(249 mg, 1.5 mmol) and DIEA (645 mg, 5 mmol) in 1,4-dioxane (20 mL)
at room temperature and the resulting mixture was stirred at
80.degree. C. for 2 h. The mixture was allowed to cool to room
temperature and then concentrated in vacuo. The residue was used in
the next step without further purification. ESI-MS m/z: 402.3
[M+H].sup.+.
tert-Butyl
2-carbamoyl-4-(7-chloro-6-(4-chlorophenyl)quinazolin-4-yl)piper-
azine-1-carboxylate
[0742] To a solution of the above obtained crude product
4-(7-chloro-6-(4-chlorophenyl)quinazolin-4-yl)piperazine-2-carboxamide
in DCM (20 mL) at room temperature, Et.sub.3N (152 mg, 1.5 mmol)
and di-tert-butyl dicarbonate (262 mg, 1.2 mmol) were added. The
mixture was stirred at room temperature for 3 h. The mixture was
concentrated in vacuo and the residue was purified by flash column
chromatography on silica gel (DCM/MeOH=30:1) to afford the desired
product (60 mg, 12% yield) as a solid. ESI-MS m/z: 502.4
[M+H].sup.+.
tert-Butyl
4-(7-chloro-6-(4-chlorophenyl)quinazolin-4-yl)-2-cyanopiperazin-
e-1-carboxylate
[0743] To a solution of tert-butyl
2-carbamoyl-4-(7-chloro-6-(4-chlorophenyl)quinazolin-4-yl)piperazine-1-ca-
rboxylate (60 mg, 0.12 mmol) and Et.sub.3N (48 mg, 0.48 mmol) in
DCM (20 mL) at 0.degree. C., TFAA (50 mg, 0.24 mmol) and the
resulting mixture was stirred at room temperature for 1 h. The
reaction mixture was quenched with saturated NaHCO.sub.3 solution,
and then extracted with DCM. The organic layer was washed with
saturated NaHCO.sub.3 solution and brine, dried over
Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was
purified by flash column chromatography on silica gel
(DCM/MeOH=50:1) to afford the desired product (50 mg, 86% yield) as
a solid. ESI-MS m/z: 484.4 [M+H].sup.+.
1-Acryloyl-4-(7-chloro-6-(4-chlorophenyl)quinazolin-4-yl)piperazine-2-carb-
onitrile
[0744] The title compound was prepared from tert-butyl
4-(7-chloro-6-(4-chlorophenyl)quinazolin-4-yl)-2-cyanopiperazine-1-carbox-
ylate according to the procedure described in steps 5 and 6 in
Example 2. .sup.1H NMR (400 MHz, DMSO-d6) .delta.: 8.7 (s, 1H), 8.1
(s, 1H), 8.0 (d, J=2.0 Hz, 1H), 7.5 (m, 4H), 6.8 (dd, J=10.4, 16.8
Hz, 1H), 6.3 (dd, J=1.6, 16.8 Hz, 1H), 5.8 (dd, J=1.6, 10.4 Hz,
1H), 4.6 (m, 1H), 4.3 (m, 3H), 3.6 (m, 2H), 3.4 (s, 1H). ESI-MS
m/z: 438.25 [M+H].sup.+.
Example 25
Synthesis of
1-(4-(7-chloro-6-(4-chlorophenyl)-2-methylquinazolin-4-yl)piperazin-1-yl)-
prop-2-en-1-one (I-50)
##STR00639##
[0745] Compound I-50 was prepared according to the general
procedures of Method M as described below:
6-Bromo-7-chloro-2-methylquinazolin-4-ol
[0746] To a solution of methyl 2-amino-5-bromo-4-chlorobenzoate
(1.0 g, 3.781 mmol) in MeCN (35 mL) at RT, dry hydrogen chloride
was added continuously for 20 min. The resulting mixture was
stirred at reflux for 2 h. The mixture was allowed to cool to RT
and poured into saturated NaHCO.sub.3 solution. The white solid was
filtered, and the filtrate was extracted with ethyl acetate. The
filtrate cake and organic layer was combined and dried over
Na.sub.2SO.sub.4, concentrated in vacuo to afford the crude product
(1.62 g) as a white solid. ESI-MS m/z: 273.3 [M+H].sup.+.
6-Bromo-4,7-dichloro-2-methylquinazoline
[0747] The mixture of 6-bromo-7-chloro-2-methylquinazolin-4-ol (500
mg, 1.828 mmol) in 30 mL of SOCl.sub.2 was stirred at reflux for 16
h. The mixture was allowed to cool to RT and concentrated in vacuo.
The residue was purified through silica chromatography (5-10% ethyl
acetate/petroleum ether) to afford the desired product (180 mg, 34%
yield) as a yellow solid.
tert-Butyl
4-(6-bromo-7-chloro-2-methylquinazolin-4-yl)piperazine-1-carbox-
ylate
[0748] To a solution of tert-butyl piperazine-1-carboxylate (76 mg,
0.410 mmol) in i-PrOH (10 mL) at RT,
6-bromo-4,7-dichloro-2-methylquinazoline (60 mg, 0.205 mmol) was
added. The resulting mixture was stirred at reflux for 40 min. The
mixture was allowed to cool to RT and partitioned between water and
ethyl acetate. The organic layer was washed with saturated
NaHCO.sub.3 and brine, dried over Na.sub.2SO.sub.4 and
concentrated. The residue was purified by flash column
chromatography on silica gel (5% ethyl acetate/petroleum ether) to
afford the desired product (53 mg, 59% yield) as a yellow
solid.
1-(4-(7-Chloro-6-(4-chlorophenyl)-2-methylquinazolin-4-yl)piperazin-1-yl)p-
rop-2-en-1-one
[0749] The title compound was prepared from tert-butyl
4-(6-bromo-7-chloro-2-methylquinazolin-4-yl)piperazine-1-carboxylate
in three steps according to the procedure described in Example 3.
.sup.1H NMR (400 MHz, DMSO-d6) .delta.: 7.92 (s, 2H), 7.59 (m, 4H),
6.84-6.77 (dd, J=10.4, 16.8 Hz, 1H), 6.17-6.36 (m, 1H), 5.74-5.71
(m, 1H), 3.85-3.72 (m, 8H), 2.54 (s, 3H). ESI-MS m/z: 428.3
[M+H].sup.+.
Example 26
Synthesis of
1-acryloyl-4-(7-chloro-6-(4-chlorophenyl)-2-methylquinazolin-4-yl)piperaz-
ine-2-carbonitrile (I-56)
##STR00640## ##STR00641##
[0750] Compound I-56 was prepared according to the general
procedures of Method M as described below:
1-tert-butyl 2-methyl
4-(6-Bromo-7-chloro-2-methylquinazolin-4-yl)piperazine-1,2-dicarboxylate
[0751] To a solution of 6-bromo-4,7-dichloro-2-methylquinazoline
(435 mg, 1.49 mmol) and 1-tert-butyl 2-methyl
piperazine-1,2-dicarboxylate (437 mg, 1.79 mmol) in 1,4-dioxane (30
mL), DIEA (769 mg, 5.96 mmol) was added. The mixture was stirred at
80.degree. C. for 1.5 h. The mixture was allowed to cool to RT and
partitioned between water and ethyl acetate. The organic layer was
washed with brine, dried over Na.sub.2SO.sub.4 and concentrated.
The residue was purified by flash column chromatography on silica
gel (5-50% ethyl acetate/petroleum ether) to afford the desired
product (224 mg, 30% yield) as a yellow solid.
4-(6-Bromo-7-chloro-2-methylquinazolin-4-yl)-1-(tert-butoxycarbonyl)pipera-
zine-2-carboxylic acid
[0752] To a solution of 1-tert-butyl 2-methyl
4-(6-bromo-7-chloro-2-methylquinazolin-4-yl)piperazine-1,2-dicarboxylate
(224 mg, 0.448 mmol) in THF (15 mL) and H.sub.2O (5 mL),
LiOH.H.sub.2O (114 mg, 2.690 mmol) was added and the resulting
mixture was stirred at RT for 1 h. The mixture was diluted with
H.sub.2O, acidified with HCl to adjust pH to 4 and then extracted
with ethyl acetate. The organic layer was washed with brine, dried
over Na.sub.2SO.sub.4 and concentrated in vacuo to afford the
desired product (211 mg, 97% yield) as a yellow solid.
tert-Butyl
4-(6-bromo-7-chloro-2-methylquinazolin-4-yl)-2-carbamoylpiperaz-
ine-1-carboxylate
[0753] To a solution of
4-(6-bromo-7-chloro-2-methylquinazolin-4-yl)-1-(tert-butoxycarbonyl)piper-
azine-2-carboxylic acid (221 mg, 0.435 mmol) and Et.sub.3N (176 mg,
1.738 mmol) in THF (35 mL) at -5.degree. C., ethyl chloroformate
(51 mg, 0.465 mmol) was added. The mixture was stirred at
-5.degree. C. for 40 min and NH.sub.3.H.sub.2O (30%, 507 mg, 4.346
mmol) was added. The resulting mixture was kept stirring for 5 min
at 0.degree. C. The mixture was partitioned between water and ethyl
acetate. The organic layer was washed with brine, dried over
Na.sub.2SO.sub.4 and concentrated. The residue was purified by
flash column chromatography on silica gel (3%
methanol/dichloromethane) to afford the desired product (179 mg,
85% yield) as a yellow solid. ESI-MS m/z: 484.3 [M+H].sup.+.
tert-Butyl
2-carbamoyl-4-(7-chloro-6-(4-chlorophenyl)-2-methylquinazolin-4-
-yl)piperazine-1-carboxylate
[0754] A mixture of tert-butyl
4-(6-bromo-7-chloro-2-methylquinazolin-4-yl)-2-carbamoylpiperazine-1-carb-
oxylate (179 mg, 0.371 mmol), (4-chlorophenyl)boronic acid (67 mg,
0.426 mmol), Pd(PPh.sub.3).sub.4(51 mg, 0.0445 mmol) and
Na.sub.2CO.sub.3 (118 mg, 1.113 mmol) in 1,4-dioxane (25 mL) was
stirred at 85.degree. C. for 16 h under argon. The mixture was
allowed to cool to RT and concentrated in vacuo. The residue was
purified by flash column chromatography on silica gel (3%
methanol/dichloromethane) to afford the desired product (181 mg,
95% yield) as a brown solid. ESI-MS m/z: 517.4 [M+H].sup.+.
tert-Butyl
4-(7-chloro-6-(4-chlorophenyl)-2-methylquinazolin-4-yl)-2-cyano-
piperazine-1-carboxylate
[0755] To a solution of tert-butyl
2-carbamoyl-4-(7-chloro-6-(4-chlorophenyl)-2-methylquinazolin-4-yl)pipera-
zine-1-carboxylate (100 mg, 0.194 mmol) and Et.sub.3N (78 mg, 0.775
mmol) in DCM (30 mL) at 0.degree. C., TFAA (162 mg, 0.776 mmol) was
added and the resulting mixture was stirred at RT for 1 h. The
reaction mixture was quenched with saturated NaHCO.sub.3 solution,
and then extracted with dichloromethane. The organic layer was
washed with saturated NaHCO.sub.3 solution and brine, dried over
Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was
purified by flash column chromatography on silica gel (petroleum
ether/ethyl acetate=2:1) to afford the desired product (58 mg, 60%
yield) as a yellow solid. ESI-MS m/z: 499.4[M+H].sup.+.
1-Acryloyl-4-(7-chloro-6-(4-chlorophenyl)-2-methylquinazolin-4-yl)piperazi-
ne-2-carbonitrile
[0756] tert-Butyl
4-(7-chloro-6-(4-chlorophenyl)-2-methylquinazolin-4-yl)-2-cyanopiperazine-
-1-carboxylate (100 mg, 0.194 mmol) was dissolved in 20 mL of 20%
HCl/Et.sub.2O solution. The mixture was stirred at RT for 30 min
and then concentrated in vacuo to yield a solid salt (44 mg, 87%
yield). The above solid (44 mg, 0.101 mmol) was dissolved in 25 mL
of DCM with Et.sub.3N (51 mg, 0.505 mmol). The mixture was cooled
to 0.degree. C. and then a solution of acryloyl chloride (10 mg,
0.111 mmol) in dichloromethane (2 mL) was added. The resulting
mixture was stirred at 0.degree. C. for 40 min. The mixture was
extracted with ethyl acetate. The organic layer was washed with
saturated NaHCO.sub.3 and brine, dried over Na.sub.2SO.sub.4 and
concentrated. The residue was purified with silica chromatography
(petroleum ether/ethyl acetate=2:1) to afford the desired product
(24 mg, 52% yield) as a white solid. .sup.1H NMR (400 MHz, DMSO-d6)
.delta.: 8.01 (d, J=6.4 Hz, 2H), 7.63 (q, J=8.4, 20.4 Hz, 4H), 6.90
(dd, J=10.4, 16.4 Hz, 1H), 6.30 (m, 1H), 5.68 (s, 1H), 4.60 (m,
1H), 4.32 (m, 2H), 3.57 (m, 2H), 2.59 (s, 3H), 3.36 (m, 1H). ESI-MS
m/z: 453.3 [M+H].sup.+.
Example 27
Synthesis of
1-(4-(7-chloro-6-(4-chlorophenyl)quinazolin-4-yl)-2-(2-hydroxyethyl)piper-
azin-1-yl)prop-2-en-1-one (I-62)
##STR00642##
[0757] Compound I-62 was prepared according to the general
procedures of Method A as described below:
Methyl 2-(3-oxopiperazin-2-yl)acetate
[0758] To a solution of dimethyl maleate (4.0 g, 27.78 mmol) in
propan-2-ol (40 mL) at RT, ethane-1,2-diamine (1.167 g, 27.78 mmol)
was added. The resulting mixture was stirred at 55.degree. C. for
16 h and concentrated in vacuo. The residue was washed by a mixture
of ethyl acetate/petroleum ether=1:1 to afford the desired product
(2.8 g, 59% yield) as a white solid.
2-(Piperazin-2-yl)ethanol
[0759] To a solution of methyl 2-(3-oxopiperazin-2-yl)acetate (1.82
g, 10.58 mmol) in THF (150 mL) at 0.degree. C., LiAlH.sub.4 (2.01
g, 52.9 mmol) was added. The resulting mixture was stirred at
reflux for 16 h. Then the mixture was cooled to RT. It was quenched
with 10H.sub.2O.Na.sub.2SO.sub.4 and filtered, washed with ethyl
acetate. The filtrated was dried over Na.sub.2SO.sub.4 and
concentrated in vacuo to afford the desired product (674 mg, 49%
yield) as a yellow oil.
2-(4-(7-Chloro-6-(4-chlorophenyl)quinazolin-4-yl)piperazin-2-yl)ethanol
[0760] A mixture of 4,7-dichloro-6-(4-chlorophenyl)quinazoline (150
mg, 0.48 mmol), 2-(piperazin-2-yl)ethanol (187 mg, 1.44 mmol),
Et.sub.3N (0.33 mL, 2.4 mmol), in 1,4-dioxane (5 mL) was stirred at
80.degree. C. for 30 min. The mixture was allowed to cool to RT,
quenched with saturated NaHCO.sub.3 solution and then extracted
with ethyl acetate. The organic layer was washed with brine, dried
over Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The
residue was purified by flash column chromatography on silica gel
(methanol/dichloroethane=1:30) to afford the desired product (121
mg, 63% yield) as a colorless oil. ESI-MS m/z: 403.3
[M+H].sup.+.
1-(4-(7-Chloro-6-(4-chlorophenyl)quinazolin-4-yl)-2-(2-hydroxyethyl)pipera-
zin-1-yl)prop-2-en-1-one
[0761] To a solution of
2-(4-(7-chloro-6-(4-chlorophenyl)quinazolin-4-yl)piperazin-2-yl)ethanol
(123 mg, 0.305 mmol), acrylic acid (24 mg, 0.336 mmol), BOP (270
mg, 0.61 mmol) in DMF (5 mL) at -30.degree. C., DIEA (157 mg, 1.22
mmol) was added. The resulting mixture was warmed to 0.degree. C.
over 1 h, quenched with saturated NaHCO.sub.3 solution, and then
extracted with ethyl acetate. The organic layer was washed with
saturated NaHCO.sub.3 solution and brine, dried over
Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was
purified by Pre-HPLC to afford the desired product (16 mg, 12%
yield) as a light-yellow oil. .sup.1H NMR (400 MHz, DMSO-d6)
.delta.: 8.64 (s, 1H), 8.01 (s, 1H), 7.99 (s, 1H), 7.64-7.57 (m,
4H), 6.89-6.78 (m, 1H), 6.17-6.13 (m, 1H), 5.72 (dd, J=2.4, 10.4
Hz, 1H), 4.72-4.58 (m, 2H), 4.38-4.29 (m, 4H), 4.06-3.99 (m, 1H),
3.67-3.60 (m, 2H), 1.79-1.68 (m, 2H). ESI-MS m/z: 457.4
[M+H].sup.+.
Example 28
Synthesis of
2-(1-acryloyl-4-(7-chloro-6-(4-chlorophenyl)quinazolin-4-yl)piperazin-2-y-
l)acetonitrile (I-70)
##STR00643##
[0762] Compound I-70 was prepared according to the general
procedures of Method A as described below:
Dibenzyl 2-(2-hydroxyethyl)piperazine-1,4-dicarboxylate
[0763] To a solution of 2-(piperazin-2-yl)ethanol (2.0 g, 15.4
mmol) in THF (48 mL), H.sub.2O (32 mL) and saturated NaHCO.sub.3
(32 mL) at 0.degree. C., Cbz-Cl (5.5 g, 32.3 mmol) was added
dropwise. The mixture was stirred at 0.degree. C. for 2 h and at RT
for 16 h. The mixture was diluted with brine, extracted with
dichloromethane. The organic layer was washed with brine, dried
over Na.sub.2SO.sub.4 and concentrated. The residue was purified by
flash column chromatography on silica gel (25%-50% ethyl
acetate/petroleum ether) to afford the desired product (1.454 g,
23% yield) as a colorless oil. ESI-MS m/z: 399.4 [M+H].sup.+.
2-(1,4-Bis((benzyloxy)carbonyl)piperazin-2-yl)acetic acid
[0764] To a solution of dibenzyl
2-(2-hydroxyethyl)piperazine-1,4-dicarboxylate (515 mg, 1.294 mmol)
in acetone (30 mL), Jones reagent (1.48 mL, 3.88 mmol, 2.6 M) was
added dropwise at 0.degree. C., which was stirred at RT for 1 h.
The mixture was quenched with i-PrOH (2 mL) and filtered through
celite. The filtrate was extracted with ethyl acetate. The organic
layer was washed with brine, dried over Na.sub.2SO.sub.4 and
concentrated to afford the crude product (545 mg) as a colorless
oil. ESI-MS m/z: 413.2 [M+H].sup.+.
Dibenzyl 2-(2-amino-2-oxoethyl)piperazine-1,4-dicarboxylate
[0765] To a solution of
2-(1,4-bis((benzyloxy)carbonyl)piperazin-2-yl)acetic acid (545 mg,
1.323 mmol) and Et.sub.3N (535 mg, 5.292 mmol) in THF (20 mL),
ethyl chloroformate (154 mg, 1.415 mmol) was added at -10 OC and
stirred at this temperature for 40 min. Then the mixture was added
NH.sub.3.H.sub.2O (1.984 g, 15.87 mmol) at -10.degree. C. and
stirred for 20 min at -10.degree. C. The mixture was partitioned
between water and ethyl acetate. The organic layer was washed with
brine, dried over Na.sub.2SO.sub.4 and concentrated. The residue
was purified by flash column chromatography on silica gel (2%
methanol/dichloromethane) to afford the desired product (393 mg,
72% yield) as a colorless oil. ESI-MS m/z: 412.3 [M+H].sup.+.
2-(Piperazin-2-yl)acetamide
[0766] A mixture of dibenzyl
2-(2-amino-2-oxoethyl)piperazine-1,4-dicarboxylate (385 mg, 0.937
mmol), Pd/C (10%, 40 mg) and MeOH (30 mL) was stirred at 40.degree.
C. for 2.5 h under H.sub.2 (1 atm). The mixture was filtered
through celite and concentrated to afford the crude product (188
mg) as a colorless oil.
2-(4-(7-Chloro-6-(4-chlorophenyl)quinazolin-4-yl)piperazin-2-yl)acetamide
[0767] A mixture of 4,7-dichloro-6-(4-chlorophenyl)quinazoline (313
mg, 1.315 mmol), 2-(piperazin-2-yl)acetamide (188 mg, 1.315 mmol),
DIEA (848 mg, 6.575 mmol) and 1,4-dioxane (30 mL) at 100.degree. C.
for 5 h. The mixture was allowed to cool to RT and concentrated in
vacuo. The residue was purified by flash column chromatography on
silica gel (5-20% methanol/dichloromethane) to afford the desired
product (78 mg, 14% yield) as a brown solid. ESI-MS m/z: 417.3
[M+H].sup.+.
2-(1-Acryloyl-4-(7-chloro-6-(4-chlorophenyl)quinazolin-4-yl)piperazin-2-yl-
)acetamide
[0768] A mixture of
2-(4-(7-chloro-6-(4-chlorophenyl)quinazolin-4-yl)piperazin-2-yl)acetamide
(78 mg, 0.1875 mmol), Et.sub.3N (76 mg, 0.750 mmol) and
dichloromethane (30 mL) at 0.degree. C., a solution of acryloyl
chloride (21 mg, 0.225 mmol) in dichloromethane (2 mL) was added
dropwise. The resulting mixture was stirred at 0.degree. C. for 40
min. The mixture was quenched with saturated NaHCO.sub.3 and
extracted with ethyl acetate. The organic layer was washed with
brine, dried over Na.sub.2SO.sub.4 and concentrated. The residue
was purified with column chromatography on silica gel (2.5-4%
methanol in dichloromethane) to afford the desired product (32 mg,
36% yield) as a white solid. .sup.1H NMR (400 MHz, DMSO-d6)
.delta.: 8.74 (s, 1H), 8.07 (s, 1H), 7.80 (s, 1H), 7.50-7.42 (dd,
J=8.8, 14.4 Hz, 1H), 6.79-6.24 (m, 3H), 5.83 (m, 1H), 5.36-5.14 (m,
2H), 4.72-4.49 (m, 2H, 4.32 (m, 1H), 3.99-3.49 (m, 3H), 3.07-2.44
(m, 3H). ESI-MS m/z: 470.2 [M+H].sup.+.
2-(1-Acryloyl-4-(7-chloro-6-(4-chlorophenyl)quinazolin-4-yl)piperazin-2-yl-
)acetonitrile
[0769] To a solution of
2-(1-acryloyl-4-(7-chloro-6-(4-chlorophenyl)quinazolin-4-yl)piperazin-2-y-
l)acetamide (25 mg, 0.0533 mmol) and Et.sub.3N (27 mg, 0.267 mmol)
in DCM (10 mL) at 0.degree. C., TFAA (46 mg, 0.214 mmol) and the
resulting mixture was stirred at RT for 20 min. The reaction
mixture was quenched with saturated NaHCO.sub.3 solution, and then
extracted with dichloromethane. The organic layer was washed with
saturated NaHCO.sub.3 solution and brine, dried over
Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was
purified by flash column chromatography on silica gel (2.5%
methanol in dichloromethane) to afford the desired product (21 mg,
87% yield) as a white solid. .sup.1H NMR (400 MHz, DMSO-d6)
.delta.: 8.67 (s, 1H), 8.06 (m, 2H), 7.70 (s, 4H), 6.88 (m, 1H),
6.20 (d, J=10.0 Hz, 1H), 5.76 (s, 1H), 4.97 (m, 1H), 4.30 (m, 4H),
3.75 (m, 2H), 2.99 (m, 2H). ESI-MS m/z: 453.3 [M+H].sup.+.
Example 29
Synthesis of
4-(4-acryloyl-3-cyanopiperazin-1-yl)-7-chloroquinazoline-6-carbonitrile
(53)
##STR00644## ##STR00645##
[0770] Compound I-53 was prepared according to the general
procedures of Method B as described below:
1-tert-Butyl 2-methyl
4-(6-bromo-7-chloroquinazolin-4-yl)piperazine-1,2-dicarboxylate
[0771] A mixture of 6-bromo-4,7-dichloroquinazoline (300 mg, 1.08
mmol), tert-butyl methyl piperazine-1,2-dicarboxylate (395 mg, 1.62
mmol), DIEA (836 mg, 6.48 mmol) in 1,4-dioxane (8 mL) was stirred
at 80.degree. C. for 1 h. The mixture was allowed to cool to RT,
quenched with saturated NaHCO.sub.3 solution and then extracted
with ethyl acetate. The organic layer was washed with brine, dried
over Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The
residue was purified by flash column chromatography on silica gel
(ethyl acetate/petroleum ether=1:5) to afford the desired product
(367 mg, 70% yield) as a white solid.
1-(tert-Butoxycarbonyl)-4-(6-bromo-7-chloroquinazolin-4-yl)piperazine-2-ca-
rboxylic acid
[0772] To a solution of 1-tert-butyl 2-methyl
4-(6-bromo-7-chloroquinazolin-4-yl)piperazine-1,2-dicarboxylate
(100 mg, 0.206 mmol) in THF (2 mL), MeOH (2 mL) and water (2 mL),
LiOH.H.sub.2O (165 mg, 4.12 mmol) was added and the resulting
mixture was stirred at RT for 1 h. The mixture was washed with 20%
ethyl acetate/petroleum ether. The aqueous layer was acidified with
aqueous HCl (1 N) to adjust pH to 5 and extracted with ethyl
acetate. The organic layer was dried over MgSO.sub.4, filtered, and
concentrated in vacuo to afford the desired product (65 mg, 67%
yield).
tert-Butyl
4-(6-bromo-7-chloroquinazolin-4-yl)-2-carbamoylpiperazine-1-car-
boxylate
[0773] To a mixture of
1-(tert-butoxycarbonyl)-4-(6-bromo-7-chloroquinazolin-4-yl)piperazine-2-c-
arboxylic acid (65 mg, 0.14 mmol), Et.sub.3N (0.11 mL, 0.77 mmol)
in THF (4 mL) and DMF (2 mL) at 0.degree. C., ethyl chloroformate
(83 mg, 0.77 mmol) was added. The resulting mixture was stirred at
0.degree. C. for 1 h and NH.sub.3.H.sub.2O (1 mL, 15 N) was added.
Then the mixture was warmed to RT and stirred for another 1 h. It
was quenched with saturated NaHCO.sub.3 solution and then extracted
with ethyl acetate. The organic layer was washed with brine, dried
over Na.sub.2SO.sub.4, filtered and concentrated in vacuo to afford
the crude product (77 mg) as a yellow solid. ESI-MS m/z: 471.4
[M+H].sup.+.
tert-Butyl2-carbamoyl-4-(7-chloro-6-cyanoquinazolin-4-yl)piperazine-1-carb-
oxylate
[0774] A mixture of tert-butyl
4-(6-bromo-7-chloroquinazolin-4-yl)-2-carbamoylpiperazine-1-carboxylate
(200 mg, 0.43 mmol), PdCl.sub.2(dppf) (31 mg, 0.043 mmol),
Zn(CN).sub.2 (80 mg, 0.68 mmol) and DMF (20 mL) was stirred at
reflux for 5 h. The mixture was allowed to cool to room
temperature, and partitioned between ethyl acetate and water. The
organic layer was washed with brine, dried over Na.sub.2SO.sub.4
and concentrated. The residue was purified by flash column
chromatography on silica gel (1-2% methanol/dichloromethane) to
afford the desired product (140 mg, 79% yield) as a solid. ESI-MS
m/z: 417.3 [M+H].sup.+.
4-(7-Chloro-6-cyanoquinazolin-4-yl)piperazine-2-carboxamide
[0775] A solution of
tert-butyl2-carbamoyl-4-(7-chloro-6-cyanoquinazolin-4-yl)piperazine-1-car-
boxylate (140 mg, 0.34 mmol) in dichloromethane (20 mL) at RT, TFA
(2 mL) was added. The resulting mixture was stirred at RT for 2 h.
The mixture was concentrated in vacuo to afford the crude product
(100 mg) which was used directly in the next step without further
purification.
1-Acryloyl-4-(7-chloro-6-cyanoquinazolin-4-yl)piperazine-2-carboxamide
[0776] A mixture of
4-(7-chloro-6-cyanoquinazolin-4-yl)piperazine-2-carboxamide (100
mg, 0.32 mmol), Et.sub.3N (96 mg, 0.96 mmol) in dichloromethane (10
mL) at 0.degree. C., acryloyl chloride (35 mg, 0.384 mmol) was
added. The resulting mixture was stirred at RT for 0.5 h, poured
into water and then extracted with dichloromethane. The organic
layer was washed with brine, dried over Na.sub.2SO.sub.4 and
concentrated. The residue was purified by flash column
chromatography on silica gel (1-2% methanol/dichloromethane) to
afford the desired product (50 mg, 43% yield) as a solid. ESI-MS
m/z: 371.3 [M+H].sup.+.
4-(4-Acryloyl-3-cyanopiperazin-1-yl)-7-chloroquinazoline-6-carbonitrile
[0777] A mixture of
1-acryloyl-4-(7-chloro-6-cyanoquinazolin-4-yl)piperazine-2-carboxamide
(50 mg, 0.14 mmol) and Et.sub.3N (82 mg, 0.81 mmol) in DCM (10 mL)
at RT, trifluoroacetic anhydride (117.6 mg, 0.56 mmol) was added.
The resulting mixture was stirred at RT for 0.5 h and poured into
water and then extracted with dichloromethane. The organic layer
was washed with brine, dried over Na.sub.2SO.sub.4 and
concentrated. The residue was purified by flash column
chromatography on silica gel (1-3% methanol/dichloromethane) to
afford the desired product (15 mg, 32% yield). .sup.1H NMR (400
MHz, DMSO-d6) .delta.: 8.79 (s, 1H), 8.77 (s, 1H), 8.16 (s, 1H),
6.92-6.85 (m, 1H), 6.32-6.28 (m, 1H), 5.91-5.88 (m, 1H), 5.68 (s,
1H), 4.73-4.70 (d, J=14 Hz, 1H), 4.46-4.43 (d, J=13.2 Hz, 1H),
4.25-4.22 (d, J=12.8 Hz, 1H), 3.82-3.74 (m, 2H), 3.59-3.56 (m, 1H).
ESI-MS m/z: 353.2 [M+H].sup.+.
Example 30
Synthesis of
1-acryloyl-4-(7-chloro-6-cyclopropylquinazolin-4-yl)piperazine-2-carbonit-
rile (55)
##STR00646##
[0778] Compound I-55 was prepared according to the general
procedures of Method B as described below:
tert-Butyl
2-carbamoyl-4-(7-chloro-6-cyclopropylquinazolin-4-yl)piperazine-
-1-carboxylate
[0779] A mixture of tert-butyl
4-(6-bromo-7-chloroquinazolin-4-yl)-2-carbamoylpiperazine-1-carboxylate
(200 mg, 0.414 mmol), cyclopropylboronic acid (44 mg, 0.51 mmol),
K.sub.3PO.sub.4.3H.sub.2O (270 mg, 1.272 mmol), Pd(OAc).sub.2 (18
mg, 0.08 mmol) and tricyclohexyl phosphine (22 mg, 0.08 mmol) in
toluene (10 mL) and water (1 mL) was stirred at reflux under argon
for 16 h. The solvent was removed, and the residue was purified by
flash column chromatography on silica gel
(dichloromethane/methanol=50:1) to afford the desired product (100
mg, 56% yield) as a solid. ESI-MS m/z: 432.4 [M+H].sup.+.
Acryloyl-4-(7-chloro-6-cyclopropylquinazolin-4-yl)piperazine-2-carboxamide
[0780] The title compound was prepared from tert-butyl
2-carbamoyl-4-(7-chloro-6-cyclopropylquinazolin-4-yl)piperazine-1-carboxy-
late in two steps following the procedure described in Example
1.
Acryloyl-4-(7-chloro-6-cyclopropylquinazolin-4-yl)piperazine-2-carboxamide
[0781] To a solution of
1-acryloyl-4-(7-chloro-6-cyclopropylquinazolin-4-yl)piperazine-2-carboxam-
ide (17 mg, 0.044 mmol) and Et.sub.3N (18 mg, 0.176 mmol) in DCM (5
mL) at 0.degree. C., TFAA (18 mg, 0.088 mmol) was added and the
resulting mixture was stirred at RT for 1 h. The reaction mixture
was quenched with saturated NaHCO.sub.3 solution, and then
extracted with dichloromethane. The organic layer was washed with
saturated NaHCO.sub.3 solution and brine, dried over
Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was
purified by flash column chromatography on silica gel
(dichloromethane/methanol=50:1) to afford the desired product (10
mg, 62% yield) as a solid. .sup.1H NMR (400 MHz, CDCl3) .delta.:
8.8 (s, 1H), 8.0 (s, 1H), 7.7 (s, 1H), 6.6 (dd, J=10.0, 16.4 Hz,
1H), 6.5 (d, J=16.4 Hz, 1H), 6.0 (dd, J=2.0, 10.4 Hz, 1H), 6.0-5.9
(m, 1H), 4.4 (dd, J=2, 13.2 Hz, 1H), 4.3-4.1 (m, 2H), 3.9-3.8 (m,
1H), 3.3-3.1 (m, 2H), 2.4-2.3 (m, 1H), 1.2-1.1 (m, 2H), 1.0-0.9 (m,
2H). ESI-MS m/z: 368.3 [M+H].sup.+.
Example 31
Synthesis of
(s)-1-acryloyl-4-(7-chloro-6-(4-chlorophenyl)quinazolin-4-yl)piperazine-2-
-carboxamide (I-54)
##STR00647##
[0782] Compound I-54 was prepared according to the general
procedures of Method A as described below:
(S)-Methyl piperazine-2-carboxylate hydrochloride
[0783] A mixture of (S)-tert-butyl methyl
piperazine-1,3-dicarboxylate (366 mg, 1.5 mmol) and HCl in MeOH (20
mL, 2.9 M) was stirred at RT for 1 h. The mixture was concentrated
in vacuo to yield the crude product (270 mg) as a yellow solid
which was used directly in next step without further
purification.
(S)-1-tert-Butyl 2-methyl
4-(7-chloro-6-(4-chlorophenyl)quinazolin-4-yl)piperazine-1,2-dicarboxylat-
e
[0784] To the mixture of above obtained crude (S)-methyl
piperazine-2-carboxylate hydrochloride,
4,7-dichloro-6-(4-chlorophenyl)quinazoline (310 mg, 1 mmol), DIEA
(1.29 g, 10 mmol) and 1,4-dioxane (20 mL) was stirred for 1 h at
80.degree. C. Then mixture was cooled to RT and di-tert butyl
dicarbonate (327 mg, 1.5 mmol) was added. The mixture was stirred
for 16 h and quenched with saturated NaHCO.sub.3 solution and then
extracted with ethyl acetate. The organic layer was washed with
brine, dried over Na.sub.2SO.sub.4, filtered and concentrated in
vacuo. The residue was purified by flash column chromatography on
silica gel (methanol/dichloroethane=1:50) to afford the desired
product (300 mg, 58% yield, 2 steps) as a solid oil. ESI-MS m/z:
517.5 [M+H].sup.+.
(S)-1-(tert-Butoxycarbonyl)-4-(7-chloro-6-(4-chlorophenyl)quinazolin-4-yl)-
piperazine-2-carboxylic acid
[0785] To a solution of (S)-1-tert-butyl 2-methyl
4-(7-chloro-6-(4-chlorophenyl)quinazolin-4-yl)piperazine-1,2-dicarboxylat-
e (300 mg, 0.58 mmol) in mixture of 1:1 tetrahydrofuran and water
(20 mL) at RT, LiOH.H.sub.2O (49 mg, 1.16 mmol) were added and the
resulting mixture was stirred for 1 h and then acidified with
aqueous HCl (1 N) to adjust the pH to 3-5. The mixture was
extracted with ethyl acetate. The combined organic layer was washed
with brine, dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated in vacuo to afford the crude product (230 mg) which
was used directly in the next step without further
purification.
(S)-tert-Butyl
2-carbamoyl-4-(7-chloro-6-(4-chlorophenyl)quinazolin-4-yl)piperazine-1-ca-
rboxylate
[0786] To a mixture of
(S)-1-(tert-butoxycarbonyl)-4-(7-chloro-6-(4-chlorophenyl)quinazolin-4-yl-
)piperazine-2-carboxylic acid (230 mg, 0.46 mmol), Et.sub.3N (139
mg, 1.37 mmol) in THF (5 mL) at 0.degree. C., ethyl chloroformate
(148 mg, 1.37 mmol) was added. The resulting mixture was stirred at
0.degree. C. for 1 h, then Ammonium hydroxide (1 mL, 15 N) was
added and kept stirring for another 1 h at RT. The mixture was
extracted with ethyl acetate dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated in vacuo. The residue was purified by
flash column chromatography on silica gel
(dichloromethane/methanol=50:1) to afford the desired product (150
mg, 65% yield) as a solid. ESI-MS m/z: 502.4 [M+H].sup.+.
(S)-1-Acryloyl-4-(7-chloro-6-(4-chlorophenyl)quinazolin-4-yl)piperazine-2--
carboxamide
[0787] The title compound was prepared from (S)-tert-butyl
2-carbamoyl-4-(7-chloro-6-(4-chlorophenyl)quinazolin-4-yl)piperazine-1-ca-
rboxylate in 2 steps according to the procedure described in
Example 2. .sup.1H NMR (400 MHz, DMSO-d6) .delta.: 8.7 (s, 1H), 8.3
(d, J=8.0 Hz, 1H), 8.0 (s, 1H), 7.8-7.5 (m, 5H), 7.4-7.2 (m, 1H),
6.9-6.6 (m, 1H), 6.2 (d, J=2.4, 17.6 Hz, 1H), 5.8-5.7 (m, 1H),
5.0-4.8 (m, 1H), 4.7 (d, J=13.2 Hz, 1H), 4.2-4.0 (m, 2H), 3.9-3.8
(m, 1H), 3.7-3.5 (m, 1H), 3.5-3.4 (m, 1H). ESI-MS m/z: 456.3
[M+H].sup.+.
Example 32
Synthesis of
(S)-1-acryloyl-4-(7-chloro-6-(4-chlorophenyl)quinazolin-4-yl)piperazine-2-
-carbonitrile (I-59)
##STR00648##
[0788] Compound I-59 was prepared according to the general
procedures of Method A as described below:
(S)-1-Acryloyl-4-(7-chloro-6-(4-chlorophenyl)quinazolin-4-yl)piperazine-2--
carbonitrile
[0789] To a solution of
(S)-1-acryloyl-4-(7-chloro-6-(4-chlorophenyl)quinazolin-4-yl)piperazine-2-
-carboxamide (23 mg, 0.05 mmol) and Et.sub.3N (20 mg, 0.2 mmol) in
DCM (5 mL) at 0.degree. C., trifluoroacetic anhydride (21 mg, 0.1
mmol) and the resulting mixture was stirred at RT for 1 h. The
reaction mixture was quenched with saturated NaHCO.sub.3 solution,
and then extracted with dichloromethane. The organic layer was
washed with saturated NaHCO.sub.3 solution and brine, dried over
Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was
purified by flash column chromatography on silica gel
(dichloromethane/methanol=50:1) to afford the desired product (15
mg, 68% yield) as a solid. .sup.1H NMR (400 MHz, DMSO-d6) .delta.:
8.7 (s, 1H), 8.1 (s, 1H), 8.0 (s, 1H), 7.5 (m, 4H), 6.8 (dd,
J=10.4, 16.4 Hz, 1H), 6.3 (dd, J=2.0, 17.2 Hz, 1H), 5.8 (dd, J=2.0,
10.8 Hz, 1H), 5.7 (m, 1H), 4.6 (d, J=14.0 Hz, 3H), 4.3 (m, 2H), 3.6
(m, 2H). ESI-MS m/z: 438.3 [M+H].sup.+.
Example 33
Synthesis of
(s)-1-(4-(7-chloro-6-(4-chlorophenyl)quinazolin-4-yl)-2-(hydroxymethyl)pi-
perazin-1-yl)prop-2-en-1-one (I-63)
##STR00649##
[0790] Compound I-63 was prepared according to the general
procedures of Method A as described below:
(S)-tert-Butyl
4-(7-chloro-6-(4-chlorophenyl)quinazolin-4-yl)-2-(hydroxymethyl)piperazin-
e-1-carboxylate
[0791] To a solution of (S)-1-tert-butyl 2-methyl
4-(7-chloro-6-(4-chlorophenyl)quinazolin-4-yl)piperazine-1,2-dicarboxylat-
e (200 mg, 0.387 mmol) in EtOH (10 mL) was added CaCl.sub.2 (215
mg, 1.933 mmol) and NaBH.sub.4 (74 mg, 1.933 mmol) at 0.degree. C.
The mixture was stirred at RT for 16 h. The mixture was filtered,
and washed by ethanol. The mixture was concentrated in vacuo, and
the residue was purified by flash column chromatography on silica
gel (dichloromethane/methanol=50:1) to afford the desired product
(80 mg, 42% yield) as a solid. ESI-MS m/z: 489.4 [M+H].sup.+.
1-((S)-4-(7-Chloro-6-(4-chlorophenyl)quinazolin-4-yl)-2-(hydroxymethyl)pip-
erazin-1-yl)prop-2-en-1-one
[0792] The title compound was prepared from (S)-tert-butyl
4-(7-chloro-6-(4-chlorophenyl)quinazolin-4-yl)-2-(hydroxymethyl)piperazin-
e-1-carboxylate in two steps according to the procedure described
in Example 14. .sup.1H NMR (400 MHz, DMSO-d6) .delta.: 8.7 (s, 1H),
8.3-8.1 (m, 1H), 8.0 (s, 1H), 7.7-7.5 (m, 4H), 6.8 (dd, J=10.4,
16.4 Hz, 1H), 6.1 (d, J=16 Hz, 1H), 5.8 (dd, J=2, 10.4 Hz, 1H),
5.1-4.9 (m, 1H), 4.3-4.1 (m, 4H), 4.2 (m, 2H), 3.7-3.5 (m, 4H).
ESI-MS m/z: 443.3 [M+H].sup.+.
Example 34
Synthesis of
1-(4-(6-chloro-7-phenylquinazolin-4-yl)piperazin-1-yl)prop-2-en-1-one
(I-67)
##STR00650##
[0793] Compound I-67 was prepared according to the general
procedures of Method B as described below:
7-Bromo-6-chloroquinazolin-4-ol
[0794] To a solution of 2-amino-4-bromo-5-chlorobenzoic acid (500
mg, 2 mmol) in EtOH (20 mL) at RT, formamidine acetate (620 mg, 6
mmol) was added. The mixture was reflux for 16 hour. The mixture
was concentrated in vacuo, and the residue was washed by saturated
NaHCO.sub.3 aqueous solution, and a mixture of ethyl
acetate/petroleum ether=1:2. The solid was dried in vacuo to get
the product (520 mg, 100% yield) which was used directly in next
step without further purification. ESI-MS m/z: 259.0
[M+H].sup.+.
7-Bromo-4,6-dichloroquinazoline
[0795] To a solution of 7-bromo-6-chloroquinazolin-4-ol (520 mg, 2
mmol) in thionyl chloride (15 mL) was added one drop of DMF. The
mixture was reflux for 16 h. The mixture was concentrated in vacuo,
the residue was used directly in next step without further
purification.
1-(4-(6-Chloro-7-phenylquinazolin-4-yl)piperazin-1-yl)prop-2-en-1-one
[0796] The title compound was prepared from
7-bromo-4,6-dichloroquinazoline in four steps according to the
procedure described in Example 3. .sup.1H NMR (400 MHz, DMSO)
.delta.: 8.7 (s, 1H), 8.2 (s, 1H), 7.8 (s, 1H), 7.6-7.4 (m, 5H),
6.85 (dd, J=10.8, 16.8 Hz, 1H), 6.2 (d, J=16.8 Hz, 1H), 5.75 (d,
J=10 Hz, 1H), 3.9-3.7 (m, 8H). ESI-MS m/z: 379.3 [M+H].sup.+.
Example 35
Synthesis of
1-(4-(7-chloro-6-(4-chlorophenyl)quinazolin-4-yl)-2-((dimethylamino)methy-
l)piperazin-1-yl)prop-2-en-1-one (I-60)
##STR00651##
[0797] Compound I-60 was prepared according to the general
procedures of Method A as described below:
di-tert-Butyl 2-(dimethylcarbamoyl)piperazine-1,4-dicarboxylate
[0798] A mixture of
1,4-bis(tert-butoxycarbonyl)piperazine-2-carboxylic acid (5 g,
15.13 mmol), dimethylamine hydrochloride (1.3 g, 15.13 mmol), EDCI
(4.3 g, 22.7 mmol), HOBt (3.1 g, 22.7 mmol) and DMF (100 mL) at
0.degree. C., Et.sub.3N (4.6 g, 45.39 mmol) was added. The mixture
was then warmed to RT and kept stirring for 2 h. The reaction
mixture was poured into water, extracted with ethyl acetate, the
combined organic layer was washed with NaHCO.sub.3 solution, brine
and dried over Na.sub.2SO.sub.4 and concentrated. The residue was
washed with petroleum ether to afford the desired product (3.64 g,
67% yield).
N,N-Dimethylpiperazine-2-carboxamide dihydrochloride
[0799] A mixture of the above obtained crude of di-tert-butyl
2-(dimethylcarbamoyl)piperazine-1,4-dicarboxylate, HCl in MeOH (50
mL, 2.9 M) was stirred at RT for 1 h, evaporated the solvent to
afford the crude product (2.4 g).
N,N-Dimethyl-1-(piperazin-2-yl)methanamine
[0800] A mixture of the above obtained crude of
N,N-dimethylpiperazine-2-carboxamide dihydrochloride (2.4 g, 10.43
mmol) and THF (50 mL) at -40.degree. C., LiAlH.sub.4 (1.6 g, 41.73
mmol) was added slowly. The mixture was heated to reflux for 3 h
and cooled to RT. It was quenched with 10H.sub.2O.Na.sub.2SO.sub.4
and filtered, washed with ethyl acetate. The filtrated was dried
over Na.sub.2SO.sub.4 and concentrated in vacuo to afford the
desired product (693 mg, 47% yield).
1-(4-(7-Chloro-6-(4-chlorophenyl)quinazolin-4-yl)piperazin-2-yl)-N,N-dimet-
hylmethanamine
[0801] A mixture of N,N-dimethyl-1-(piperazin-2-yl)methanamine (200
mg, 0.68 mmol), 4,7-dichloro-6-(4-chlorophenyl)quinazoline (111 mg,
0.77 mmol), DIEA (397 mg, 3.08 mmol) and dioxane (10 mL) was
stirred at 80.degree. C. for 30 min. The mixture was allowed to
cool to RT, quenched with saturated NaHCO.sub.3 solution and then
extracted with ethyl acetate. The organic layer was washed with
brine, dried over Na.sub.2SO.sub.4, filtered and concentrated in
vacuo. The residue was purified by flash column chromatography on
silica gel (methanol/dichloroethane=1:20) to afford the desired
product (78 mg, 30% yield). ESI-MS m/z: 416.3 [M+H].sup.+.
1-(4-(7-Chloro-6-(4-chlorophenyl)quinazolin-4-yl)-2-((dimethylamino)methyl-
)piperazin-1-yl)prop-2-en-1-one
[0802] A mixture of
1-(4-(7-chloro-6-(4-chlorophenyl)quinazolin-4-yl)piperazin-2-yl)-N,N-dime-
thylmethanamine (78 mg, 0.19 mmol), Et.sub.3N (58 mg, 0.57 mmol)
and dichloromethane (15 mL) at 0.degree. C., acryloyl chloride (20
mg, 0.22 mmol) was added. The reaction was stirred at RT for 30 min
and quenched with water, extracted with dichloromethane. The
organic layer was washed with water and brine, anhydrous sodium
sulfate, filtered and concentrated in vacuo. The residue was
purified by flash column chromatography on silica gel
(dichloromethane/methanol=30:1) to afford the desired product (32
mg, 36% yield). .sup.1H NMR (400 MHz, DMSO-d6) .delta.: 8.70 (s,
1H), 8.57-8.56 (bs, 1H), 8.03 (s, 1H), 7.61-7.53 (m, 4H), 6.83-6.80
(m, 1H), 6.17-6.13 (m, 1H), 5.75-5.72 (m, 1H), 4.76-4.74 (m, 0.5H),
4.70-4.57 (m, 1H), 4.36-3.29 (m, 2H), 4.11-4.08 (m, 0.5H), 3.46 (m,
1H), 3.27-3.11 (m, 2H), 2.93-2.84 (m, 1H), 1.99-1.94 (m, 1H), 1.87
(s, 6H). ESI-MS m/z: 470.4 [M+H].sup.+.
Example 36
Synthesis of
1-acryloyl-4-(6-chloroisoquinolin-1-yl)piperazine-2-carbonitrile
(I-61)
##STR00652##
[0803] Compound I-61 was prepared according to the general
procedures of Method D as described below:
6-Chloroisoquinoline 2-oxide
[0804] To a stirred solution 6-chloroisoquinoline (1.0 g, 6.1 mmol)
in dichloromethane (20 mL) at RT, 3-chlorobenzoperoxoic acid (1.57
g, 9.2 mmol) was added. The reaction mixture was stirred at RT for
2 h. The precipitate was filtered off and washed with
dichloromethane, the filtrate was washed twice with NaHCO.sub.3
solution. The organic layer was dried with Na.sub.2SO.sub.4 and
concentrated in vacuo to afford the desired product (1.05 g, 96%
yield) as a white solid. ESI-MS m/z: 180.2 [M+H].sup.+.
1,6-Dichloroisoquinoline
[0805] A mixture of 6-chloroisoquinoline 2-oxide (1.0 g, 5.58 mmol)
and POCl.sub.3 (10 mL) was heated to reflux for 4 h. After cooled
down to RT, the reaction mixture was poured into ice-water, and
extracted with dichloromethane. The organic layer was dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo to
afford the desired crude product which was used in the next step
without further purification.
4-(6-Chloroisoquinolin-1-yl)piperazine-2-carboxamide
[0806] To a stirred solution of 1,6-dichloroisoquinoline (500 mg,
2.56 mmol) in DMSO (5 mL) at RT, piperazine-2-carboxamide (425.6
mg, 2.56 mmol) and K.sub.2CO.sub.3 (1.05 g, 7.68 mmol). The
reaction mixture was heated at 80.degree. C. for 5 h. The reaction
mixture was diluted with ethyl acetate and washed with brine. The
organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered
and concentrated in vacuo. The residue was purified by flash column
chromatography on silica gel (ethyl acetate/petroleum ether=1:5) to
afford the desired product (80 mg, 12% yield). ESI-MS m/z:
291[M+H].sup.+.
Acryloyl-4-(6-chloroisoquinolin-1-yl)piperazine-2-carboxamide
[0807] To a mixture of
4-(6-chloroisoquinolin-1-yl)piperazine-2-carboxamide (50 mg, 0.172
mmol), triethylamine (52.1 mg, 0.51 mmol) in dichloromethane (20
mL), acryloyl chloride (15.6 mg, 0.172 mmol) in dichloromethane (1
mL) was added dropwise. The reaction mixture was stirred at RT for
30 min, poured into water, and extracted with dichloromethane. The
organic layer was washed with water and brine, dried over
Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was
purified by flash column chromatography on silica gel
(dichloromethane/methanol=100:1) to afford the desired product (45
mg, 76.3% yield). ESI-MS m/z: 345 [M+H].sup.+.
Acryloyl-4-(6-chloroisoquinolin-1-yl)piperazine-2-carbonitrile
[0808] To a mixture of
1-acryloyl-4-(6-chloroisoquinolin-1-yl)piperazine-2-carboxamide (40
mg, 0.116 mmol), triethylamine (46.8 mg, 0.46 mmol) in DCM (5 mL)
at 0.degree. C., trifluoroacetic anhydride (50 mg, 0.233 mmol) was
added. The reaction mixture was warmed to RT over 1 h, poured into
water and extracted with dichloromethane. The organic layer was
washed with water and brine, dried over Na.sub.2SO.sub.4 and
concentrated in vacuo. The residue was purified by flash column
chromatography on silica gel (dichloromethane/methanol=100:1) to
afford the desired product (20 mg, 53% yield). .sup.1H NMR (400
MHz, DMSO-d6) .delta.: 8.25 (m, 1H), 8.22 (m, 1H), 8.11 (s, 1H),
7.71 (m, 1H), 7.52 (m, 1H), 6.96 (dd, J=10.5, 16.9 Hz, 1H), 6.32
(dd, J=1.7, 16.7 Hz, 1H), 5.90 (dd, J=1.7, 16.7 Hz, 1H), 5.79 (m,
1H), 4.34 (m, 1H), 3.99 (m, 1H), 3.79 (m, 1H), 3.66 (m, 1H), 3.16
(m, 1H), 2.97 (m, 1H). ESI-MS m/z: 327 [M+H].sup.+.
Example 37
Synthesis of
(e)-4-(7-chloro-6-(4-chlorophenyl)quinazolin-4-yl)-1-(4-(dimethylamino)bu-
t-2-enoyl)piperazine-2-carbonitrile (I-66)
##STR00653##
[0809] Compound I-66 was prepared according to the general
procedures of Method A as described below:
4-(7-Chloro-6-(4-chlorophenyl)quinazolin-4-yl)piperazine-2-carboxamide
[0810] A mixture of 4,7-dichloro-6-(4-chlorophenyl)quinazoline (769
mg, 2.48 mmol), piperazine-2-carboxamide dihydrochloride (498 mg,
2.48 mmol), DIPEA (3.2 g, 24.8 mmol) and 1,4-dioxane (20 mL) was
stirred at 80.degree. C. for 16 h. The mixture was allowed to cool
to RT, quenched with saturated NaHCO.sub.3 solution and then
extracted with ethyl acetate. The organic layer was washed with
brine, dried over Na.sub.2SO.sub.4, filtered and concentrated in
vacuo. The residue was purified by flash column chromatography on
silica gel (methanol/dichloroethane=1:20) to afford the desired
product (486 mg, 48.7% yield).
(E)-4-(7-Chloro-6-(4-chlorophenyl)quinazolin-4-yl)-1-(4-(dimethylamino)but-
-2-enoyl)piperazine-2-carboxamide
[0811] To a mixture of
4-(7-chloro-6-(4-chlorophenyl)quinazolin-4-yl)piperazine-2-carboxamide
(100 mg, 0.26 mmol), BOP (256.6 mg, 0.58 mmol),
(E)-4-(dimethylamino)but-2-enoic acid (48 mg, 0.58 mmol) in
dichloromethane (10 ml) at RT, DIEA (108.6 mg, 0.78 mmol) was
added. The mixture was stirred for 30 min, extracted with ethyl
acetate. The organic layer was washed with brine, dried over
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The residue
was purified by flash column chromatography on silica gel
(methanol/dichloroethane=1:10) to afford the desired product (50
mg, 39% yield). ESI-MS m/z: 513.3 [M+H].sup.+
(E)-4-(7-Chloro-6-(4-chlorophenyl)quinazolin-4-yl)-1-(4-(dimethylamino)but-
-2-enoyl)piperazine-2-carbonitrile
[0812] To a solution of
(E)-4-(7-chloro-6-(4-chlorophenyl)quinazolin-4-yl)-1-(4-(dimethylamino)bu-
t-2-enoyl)piperazine-2-carboxamide (50 mg, 0.10 mmol) and Et.sub.3N
(0.05 mL, 0.40 mmol) in DCM (10 mL) at 0.degree. C., TFAA (51 mg,
0.20 mmol) and the resulting mixture was stirred at RT for 1 h. The
reaction mixture was quenched with saturated NaHCO.sub.3 solution,
and then extracted with dichloromethane. The organic layer was
washed with saturated NaHCO.sub.3 solution and brine, dried over
Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was
purified by flash column chromatography on silica gel
(dichloromethane/methanol=20:1) to afford the desired product (14
mg, 29% yield) as a solid. .sup.1H NMR (400 MHz, DMSO-d6) .delta.:
8.76 (s, 1H), 8.08 (d, J=16 Hz, 2H), 7.61 (dd, J=8, 24 Hz, 4H),
6.78-6.72 (m, 2H), 5.67 (s, 1H), 4.62 (d, J=14.4 Hz, 1H), 4.36-4.26
(m, 2H), 3.63 (d, J=12.4 Hz, 1H), 3.21 (s, 2H), 3.03 (d, J=6.4 Hz,
2H), 2.26 (s, 1H). ESI-MS m/z: 495.4 [M+H].sup.+.
Example 38
Synthesis of
1-(3-(4-(2-(4,5-dichloro-2-hydroxyphenylamino)acetyl)piperazin-1-yl)azeti-
din-1-yl)prop-2-en-1-one
##STR00654##
[0813] tert-Butyl
3-(4-(2-(4,5-dichloro-2-hydroxyphenylamino)acetyl)piperazin-1-yl)azetidin-
e-1-carboxylate
[0814] A mixture of 2-(4,5-dichloro-2-hydroxyphenylamino)acetic
acid (500 mg, 2.12 mmol), tert-butyl
3-(piperazin-1-yl)azetidine-1-carboxylate (565 mg, 2.34 mmol),
EDCI.HCl (488 mg, 2.54 mmol), HOBt (343 mg, 2.54 mmol), Et.sub.3N
(428 mg, 4.24 mmol) in DMF (20 mL) was stirred at room temperature
for 15 h. The mixture was poured into water and extracted with
ethyl acetate. The organic layer was washed withed saturated
aqueous NaHCO.sub.3 solution and brine, dried over Na.sub.2SO.sub.4
and concentrated in vacuo. The residue was purified by flash column
chromatography on silica gel (DCM/MeOH=30:1) to afford the desired
product (300 mg, 31% yield). ESI-MS m z: 457.4 [M-H].sup.-.
2-(4,5-Dichloro-2-hydroxyphenylamino)-1-(4-(azetidin-3-yl)piperazin-1-yl)e-
thanone hydrochloride
[0815] A mixture of tert-butyl
3-(4-(2-(4,5-dichloro-2-hydroxyphenylamino)acetyl)piperazin-1-yl)azetidin-
e-1-carboxylate (150 mg, 0.33 mmol) in HCl-MeOH (20 mL, 57 mmol)
was stirred at room temperature for 1 h. The mixture was
concentrated in vacuo to afford the crude product (130 mg) which
was used directly in the next step without further
purification.
1-(3-(4-(2-(4,5-Dichloro-2-hydroxyphenylamino)acetyl)piperazin-1-yl)azetid-
in-1-yl)prop-2-en-1-one
[0816]
2-(4,5-dichloro-2-hydroxyphenylamino)-1-(4-(azetidin-3-yl)piperazin-
-1-yl)ethanone hydrochloride (120 mg, 0.30 mmol) was added to the
mixture of Et.sub.3N (0.2 mL, 1.44 mmol) in DCM (10 mL) followed by
addition of DMF (1 drop). The mixture was stirred for 5 min and
then acryloyl chloride (27 mg, 0.30 mmol) was added. The resulting
mixture was stirred at room temperature for 1 h, poured into water
and then extracted with MeOH/DCM. The organic layer was washed with
brine, dried over Na.sub.2SO.sub.4 and concentrated. The residue
was purified by flash column chromatography on silica gel
(DCM/MeOH/NH.sub.3.H.sub.2O=50:1:0.1 to 20:1:0.2) to afford the
desired product (30 mg, 24% yield). .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta.: 10.17 (s, 1H), 6.78 (s, 1H), 6.71 (s, 1H),
6.30 (dd, J=10.4, 17.2 Hz, 1H), 6.09 (dd, J=2.0, 17.2 Hz, 1H), 5.67
(dd, J=2.4, 10.4 Hz, 1H), 5.32 (t, J=4.4 Hz, 1H), 4.26-4.22 (m,
1H), 4.11-4.04 (m, 1H), 3.93-3.91 (m, 3H), 3.79-3.75 (m, 3H),
3.52-3.51 (m, 4H), 3.19-3.16 (m, 1H), 2.36-2.30 (m, 4H). ESI-MS
m/z: 411.2 [M-H].sup.-.
Example 39
Synthesis of
N-(1'-(2-(4,5-dichloro-2-hydroxyphenylamino)acetyl)-1,3'-biazetidin-3-yl)-
acrylamide
##STR00655##
[0817] tert-Butyl
3-(3-(acrylamido)azetidin-1-yl)azetidine-1-carboxylate
[0818] To a mixture of N-(azetidin-3-yl)acrylamide hydrochloride
(500 mg, 3.40 mmol), tert-butyl 3-oxoazetidine-1-carboxylate (684
mg, 4.0 mmol), Et.sub.3N (343 mg, 3.40 mmol) and AcOH (100 mg,
0.167 mmol) in DCM (20 mL), NaBH(OAc).sub.3 (2.16 g, 10.2 mmol) was
added, and the resulting mixture was stirred at room temperature
for 16 h. The mixture was poured into water and extracted with
ethyl acetate. The organic layer was washed with saturated
NaHCO.sub.3 solution and brine, dried over Na.sub.2SO.sub.4 and
concentrated in vacuo. The residue was purified by flash column
chromatography on silica gel (DCM/MeOH=100:1 to 20:1) to afford the
desired product (300 mg, 31% yield).
N-(1-(Azetidin-3-yl)azetidin-3-yl)acrylamide hydrochloride
[0819] A mixture of tert-butyl
3-(3-(acrylamido)azetidin-1-yl)azetidine-1-carboxylate (300 mg,
1.07 mmol) in HCl-MeOH (30 mL, 86 mmol) was stirred at room
temperature for 1 h. The mixture was concentrated in vacuo to
afford the crude product (250 mg) which was used directly in the
next step without further purification.
N-(1-(1-(2-(4,5-Dichloro-2-hydroxyphenylamino)acetyl)azetidin-3-yl)azetidi-
n-3-yl)acrylamide
[0820] A mixture of 2-(4,5-dichloro-2-hydroxyphenylamino)acetic
acid (120 mg, 0.51 mmol), EDCI.HCl (147 mg, 0.77 mmol), HOBt (83
mg, 0.61 mmol), Et.sub.3N (154 mg, 1.53 mmol) in DMF (20 mL) was
stirred at room temperature for 5 min and then
N-(1-(azetidin-3-yl)azetidin-3-yl)acrylamide hydrochloride (150 mg,
0.69 mmol) was added. The resulting mixture was stirred at room
temperature for 15 h. The mixture was poured into water and
extracted with ethyl acetate. The organic layer was washed with
saturated NaHCO.sub.3 solution and brine, dried over
Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was
purified by flash column chromatography on silica gel
(DCM/MeOH/NH.sub.3.H.sub.2O=100:10:1.5) to afford the desired
product (6 mg, 3% yield). .sup.1H NMR (400 MHz, DMSO-d6) .delta.:
10.19 (s, 1H), 8.59 (d, J=8.0 Hz, 1H), 6.79 (s, 1H), 6.55 (s, 1H),
6.20 (dd, J=10.0, 16.8 Hz, 1H), 6.09 (dd, J=2.0, 16.8 Hz, 1H), 5.62
(dd, J=2.0, 9.6 Hz, 1H), 5.20 (t, J=4.0 Hz, 1H), 4.40-4.35 (m, 1H),
4.19-4.15 (m, 1H), 3.96-3.88 (m, 2H), 3.73-3.69 (m, 3H), 3.53-3.45
(m, 3H), 3.00-2.96 (m, 2H). ESI-MS m/z: 399.2 [M+H].sup.+.
Example 40
Synthesis of
1-(2-(2-(4,5-dichloro-2-hydroxyphenylamino)acetyl)-2,6-diazaspiro[3.4]oct-
an-6-yl)prop-2-en-1-one
##STR00656##
[0821] 2,6-Diaza-spiro[3.4]octane-6-acryloyl-2-carboxylic acid
tert-butyl ester
[0822] To a mixture of 2,6-diaza-spiro[3.4]octane-2-carboxylic acid
tert-butyl ester (80 mg, 0.38 mmol), Et.sub.3N (0.2 mL, 1.44 mmol)
in DCM (20 mL), acryloyl chloride (34 mg, 0.38 mmol) was added and
the resulting mixture was stirred at room temperature for 1 h. The
mixture was poured into water and extracted with ethyl acetate. The
organic layer was washed with saturated NaHCO.sub.3 solution and
brine, dried over Na.sub.2SO.sub.4 and concentrated in vacuo. The
residue was purified by flash column chromatography on silica gel
(DCM/MeOH=40:1) to afford the desired product (50 mg, 50% yield).
ESI-MS m/z: 289.2 [M+Na].sup.+.
1-(2,6-Diazaspiro[3.4]octan-6-yl)prop-2-en-1-one
[0823] A mixture of
2,6-diaza-spiro[3.4]octane-6-acryloyl-2-carboxylic acid tert-butyl
ester (50 mg, 0.19 mmol) in HCl/MeOH (10 mL, 29 mmol) was stirred
at room temperature for 1 h. The mixture was concentrated in vacuo
to afford the crude product (40 mg) which was used directly in the
next step without further purification.
1-(2-((4,5-Dichloro-2-hydroxyphenyl)glycyl)-2,6-diazaspiro[3.4]octan-6-yl)-
prop-2-en-1-one
[0824] The mixture of 2-(4,5-dichloro-2-hydroxyphenylamino)acetic
acid (47 mg, 0.2 mmol),
1-(2,6-diazaspiro[3.4]octan-6-yl)prop-2-en-1-one (40 mg, 0.2 mmol),
EDCI.HCl (46 mg, 0.24 mmol), HOBt (32 mg, 0.24 mmol) and Et.sub.3N
(0.61 mg, 0.6 mmol) in DMF (10 mL) was stirred at room temperature
for 2 h. The mixture was poured into water and extracted with ethyl
acetate. The organic layer was washed withed saturated NaHCO.sub.3
solution and brine, dried over Na.sub.2SO.sub.4 and concentrated in
vacuo. The residue was purified by flash column chromatography on
silica gel (DCM/MeOH=30:1) to afford the desired product (13 mg,
17% yield). .sup.1H NMR (400 MHz, DMSO-d6) .delta.: 10.17 (s, 1H),
6.78 (s, 1H), 6.60-6.50 (m, 2H), 6.13 (dt, J=2.4, 16.4 Hz, 1H),
5.67 (dd, J=2.4, 10.4 Hz, 1H), 5.19 (dd, J=5.2, 10.0 Hz, 1H),
4.16-4.07 (m, 2H), 3.90-3.83 (m, 2H), 3.75-3.72 (m, 3H), 3.61-3.52
(m, 2H), 3.42-3.39 (m, 1H), 2.16-2.13 (m, 1H), 2.06-2.03 (m, 1H).
ESI-MS m/z: 382.3 [M-H].sup.-.
Example 41
Synthesis of
1-(4-(2'-chloro-5-hydroxybiphenylcarbonyl)piperazin-1-yl)prop-2-en-1-one
(III-25)
##STR00657##
[0825] 2-Amino-5-bromo-3-methoxybenzoic acid
[0826] To a solution of 2-amino-3-methoxybenzoic acid (5 g, 29.9
mmol) in MeOH (35 mL) at -5.degree. C., NBS (5.59 g, 31.4 mmol) was
added and the resulting mixture was stirred at 0.degree. C. for 16
h. The mixture was partitioned between ethyl acetate and water. The
organic layer was washed with brine, dried over MgSO.sub.4,
filtered and concentrated in vacuo to afford the crude product (4
g, 54% yield). ESI-MS m/z: 244.2 [M-H].sup.-.
3-Bromo-5-methoxybenzoic acid
[0827] To a solution of 2-amino-5-bromo-3-methoxybenzoic acid (4 g,
16.3 mmol) in water (20 mL) at 0.degree. C., conc. HCl (7.5 mL, 90
mmol) and THF (20 mL) were added. The mixture was stirred for 30
min, and then NaNO.sub.2 (3.16 g, 45.8 mmol) was added. The
resulting mixture was stirred for 2 h and then hypophosphorous acid
(5.1 g, 76 mmol, 50% in H.sub.2O) was added to the reaction. The
mixture was stirred at room temperature for 16 h. The precipitate
was collected by filtration, washed with water and dried in vacuo
to afford the desired product (3.2 g, 85% yield). ESI-MS m/z: 229.2
[M-H].sup.-.
3-(2-Chlorophenyl)-5-methoxybenzoic acid
[0828] To a solution of 3-bromo-5-methoxybenzoic acid (1 g, 4.06
mmol) and 2-chlorophenylboronic acid (1.27 g, 8.13 mmol) in
1,4-dioxane (10 mL) and water (2 mL), Pd(PPh.sub.3).sub.4(468 mg,
0.40 mmol) and Na.sub.2CO.sub.3 (2.15 g, 20.3 mmol) were added and
the resulting mixture was stirred at 80.degree. C. for 16 h. The
mixture was allowed to cool to room temperature and acidified with
aqueous HCl (1.0 M) to adjust the pH to 3-4. The mixture was
extracted with ethyl acetate. The organic layer was dried over
MgSO.sub.4, filtered and concentrated in vacuo to afford the
desired product (800 mg, 75% yield) without further purification.
ESI-MS m/z: 361.2 [M-H].sup.-.
1-(4-(2'-Chloro-5-methoxy-[1,1'-biphenyl]-3-carbonyl)piperazin-1-yl)prop-2-
-en-1-one
[0829] To a solution of tert-butyl
4-acryloylpiperazine-1-carboxylate (260 mg, 1.07 mmol) in DCM (2
mL), a solution of HCl in MeOH (10 mL, 28.6 mmol) was added and the
resulting mixture was stirred at room temperature for 1 h. The
mixture was concentrated in vacuo. The residue was added to the
solution of 3-(2-chlorophenyl)-5-methoxybenzoic acid (280 mg, 1.07
mmol), HOBt (290 mg, 2.17 mmol), EDCI.HCl (410 mg, 2.17 mmol) and
Et.sub.3N (324 mg, 3.21 mmol) in DMF (10 mL). The resulting mixture
was stirred at room temperature for 16 h and partitioned between
DCM and saturated NaHCO.sub.3 solution. The organic layer was dried
over MgSO.sub.4, filtered and concentrated in vacuo. The residue
was purified by flash column chromatography on silica gel
(dichloromethane/methanol=50:1) to afford the desired product (200
mg, 52% yield). ESI-MS m/z: 385.2[M+H].sup.+.
1-(4-(2'-Chloro-5-hydroxy-[1,1'-biphenyl]-3-carbonyl)piperazin-1-yl)prop-2-
-en-1-one (VI-25)
[0830] To a solution of
1-(4-(2'-chloro-5-methoxy-[1,1'-biphenyl]-3-carbonyl)piperazin-1-yl)prop--
2-en-1-one (100 mg, 0.26 mmol) in DCM (15 mL) at -78.degree. C.,
BBr.sub.3 (650 mg, 2.6 mmol) was added and the resulting mixture
was stirred at room temperature for 1 h. The mixture was poured
into ice-water, basified with sat NaHCO.sub.3 aqueous solution to
adjust the pH to 7-8 and extracted with ethyl acetate. The organic
layer was dried over MgSO.sub.4, filtered and concentrated in
vacuo. The residue was purified by flash column chromatography on
silica gel (dichloromethane/methanol=50:1) to afford the desired
product (25 mg, 26% yield). .sup.1H NMR (400 MHz, DMSO-d6) .delta.:
9.96 (s, 1H), 7.58-7.56 (m, 1H), 7.42-7.40 (m, 3H), 6.89-6.75 (m,
4H), 6.14 (dd, J=2.0, 16.8 Hz, 1H), 5.71 (dd, J=2.0, 10.0 Hz, 1H),
3.68-3.44 (m, 8H). ESI-MS m/z: 371.2 [M+H].sup.+.
Example 42
Synthesis of
1-(4-(2',6-dichloro-4-hydroxybiphenylcarbonyl)piperazin-1-yl)prop-2-en-1--
one (III-3)
##STR00658##
[0831]
1-(4-(2',6-dichloro-4-methoxy-[1,1'-biphenyl]-3-carbonyl)piperazin--
1-yl)prop-2-en-1-one
[0832] tert-Butyl
4-(2',6-dichloro-4-methoxy-[1,1'-biphenyl]-3-carbonyl)piperazine-1-carbox-
ylate (200 mg, 0.43 mmol) was stirred in HCl in MeOH (2.86 M, 10
mL) for 1 h. The mixture was concentrated in vacuo to yield the
crude product. The residue was dissolved in DCM (15 mL),
triethylamine (0.5 mL), acryloyl chloride (40 mg, 0.43 mmol) was
added to the mixture. The reaction mixture was stirred at room
temperature for 30 min, poured into water, and extracted with DCM.
The organic layer was washed with water and brine, dried over
Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was
purified by flash column chromatography on silica gel
(dichloromethane/methanol=50:1) to afford the desired product (16
mg, 10% yield) as white solid. ESI-MS m/z: 419.2 [M+H].sup.+.
1-(4-(2',6-Dichloro-4-hydroxy-[1,1'-biphenyl]-3-carbonyl)piperazin-1-yl)pr-
op-2-en-1-one (VI-3)
[0833] To a solution of
1-(4-(2',6-Dichloro-4-methoxy-[1,1'-biphenyl]-3-carbonyl)piperazin-1-yl)p-
rop-2-en-1-one (200 mg, 0.48 mmol) in DCM (15 mL) at -60.degree.
C., BBr.sub.3 (0.6 g, 2.4 mmol) was added dropwise and the
resulting mixture was stirred at room temperature for 1 h. The
mixture was poured into ice-water, basified with saturated
NaHCO.sub.3 solution to adjust the pH to 8-9, and extracted with
DCM. The organic layer was dried over anhydrous sodium sulfate,
filtered and concentrated in vacuo. The residue was purified by
flash column chromatography on silica gel
(dichloromethane/methanol=20:1) to afford the desired product (10
mg, 5% yield). .sup.1H NMR (400 MHz, DMSO-d6) .delta.: 10.6 (s,
1H), 7.57-7.33 (m, 5H), 7.12 (s, 1H), 7.05 (s, 1H), 6.80 (m, 1H),
6.15-6.11 (dd, J=2, 16.8 Hz, 1H), 5.72-5.70 (m, 1H), 3.6 (m, 8H).
ESI-MS m/z: 405.3 [M+H].sup.+.
Example 43
Synthesis of
1-(1-acryloylazetidin-3-yl)-N-(4,5-dichloro-2-hydroxybenzyl)piperidine-4--
carboxamide (II-17)
##STR00659## ##STR00660##
[0834] 4,5-Dichloro-2-methoxybenzoic acid
[0835] A mixture of 4-chloro-2-methoxybenzoic acid (10 g, 53.6
mmol) and NCS (35 g, 19.2 mmol) in acetonitrile (200 mL) was
stirred at room temperature for 48 h. The mixture was poured into
water and extracted with ethyl acetate. The organic layer was
washed with water and brine, dried over Na.sub.2SO.sub.4 and
concentrated in vacuo to get the crude product (23.3 g).
Methyl 4,5-dichloro-2-methoxybenzoate
[0836] A mixture of 4,5-dichloro-2-methoxybenzoic acid (8.2 g, 37
mmol) and K.sub.2CO.sub.3 (11.8 g, 111 mmol) in DMF (100 mL),
CH.sub.3I (6.3 g, 44 mmol) was added dropwise and the resulting
mixture was stirred at room temperature for 16 h. The mixture was
partitioned between ethyl acetate and water. The organic layer was
washed with water and brine, dried over Na.sub.2SO.sub.4, and
concentrated in vacuo. The residue was purified by flash column
chromatography on silica gel (petroleum ether/ethyl acetate=10:1)
to afford the desired product.
(4,5-Dichloro-2-methoxyphenyl)methanol
[0837] To a mixture of LiAlH.sub.4 (2.42 g, 64 mmol) in THE (40 mL)
at -40.degree. C. under argon, a solution of methyl
4,5-dichloro-2-methoxybenzoate (6 g, 26 mmol) in THF (50 mL) was
added dropwise. The reaction mixture was stirred at -5.degree. C.
to 5.degree. C. for 1 h. The mixture was cooled to -20.degree. C.
and then water (2 mL) and NaOH (15%) aqueous were added. The
resulting mixture was stirred for 15 min. The solid was filtered,
and the cake rinsed with ethyl acetate. The combined filtrate was
dried over Na.sub.2SO.sub.4 and concentrated in vacuo to afford the
crude product (4.6 g).
2-(4,5-Dichloro-2-methoxybenzyl)isoindoline-1,3-dione
[0838] To a mixture of 4,5-dichloro-2-methoxyphenyl)methanol (4.5
g, 22 mmol), isoindoline-1,3-dione (9.6 g, 65 mmol) and PPh.sub.3
(17 g, 65 mmol) in THF (100 mL) at room temperature, DIAD (13 g, 65
mmol) was added and the resulting mixture was stirred at room
temperature for 16 h. The mixture was partitioned between ethyl
acetate and water. The organic layer was washed brine, dried over
Na.sub.2SO.sub.4, concentrated in vacuo. The residue was purified
by flash column chromatography on silica gel (petroleum ether/ethyl
acetate=10:1) to afford the desired product.
(4,5-Dichloro-2-methoxyphenyl)methanamine
[0839] To a solution of
2-(4,5-dichloro-2-methoxybenzyl)isoindoline-1,3-dione (1.8 g, 5
mmol) in EtOH (5 mL), hydrazine hydrate (1.34 g, 27 mmol) was added
and the resulting mixture was stirred at reflux for 1 h. The
mixture was concentrated in vacuo and the residue was purified by
flash column chromatography on silica gel
(dichloromethane/methanol=20:1) to afford the desired product (0.8
g, 78% yield).
tert-Butyl
4-((4,5-dichloro-2-methoxybenzyl)carbamoyl)piperidine-1-carboxy-
late
[0840] The mixture of (4,5-dichloro-2-methoxyphenyl)methanamine
(0.8 g, 3.90 mmol), 1-(tert-butoxycarbonyl)piperidine-4-carboxylic
acid (0.88 g, 3.84 mmol), BOP (2 g, 1.16 mmol) and DIEA (1.6 g,
2.91 mmol) in DMF (20 mL) was stirred at room temperature for 1 h.
The mixture was poured into water and extracted with ethyl acetate.
The organic layer was washed with water and brine, dried over
Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was
purified by flash column chromatography on silica gel
(dichloromethane/methanol=100:1) to afford the desired product
(0.987 g, 62% yield). ESI-MS m/z: 415.4 [M-H].sup.-.
N-(4,5-Dichloro-2-methoxybenzyl)piperidine-4-carboxamide
[0841] The mixture of
4-((4,5-dichloro-2-methoxybenzyl)carbamoyl)piperidine-1-carboxylate
(987 mg, 2.37 mmol) in HCl/MeOH (20 mL, 57.2 mmol) was stirred at
room temperature for 1 h. Then the solvent was evaporated in vacuo
and the residue was dissolved with dichloromethane (5 mL). To this
mixture, NaH (85 mg, 3.55 mmol) was added. Then the resulting
mixture was stirred at room temperature for 30 min. The solvent was
removed under reduced pressure to yield the crude product (800
mg).
tert-butyl
3-(4-(4,5-Dichloro-2-methoxybenzylcarbamoyl)piperidin-1-yl)azet-
idine-1-carboxylate
[0842] A mixture of
N-(4,5-dichloro-2-methoxybenzyl)piperidine-4-carboxamide (750 mg,
2.37 mmol), tert-butyl 3-oxoazetidine-1-carboxylate (607 mg, 3.55
mmol), AcOH (1 mL) and MeOH (5 mL) was stirred at reflux for 2 h.
To this mixture, NaBH.sub.3(CN) (0.74 g, 11.85 mmol) was added and
the resulting mixture was stirred at 60.degree. C. for 16 h. The
mixture was allowed to cool to room temperature and partitioned
between NH.sub.4Cl aqueous solution and ethyl acetate. The organic
layer was washed with water and brine, dried over Na.sub.2SO.sub.4
and concentrated in vacuo. The residue was purified by flash column
chromatography on silica gel (dichloromethane/methanol=50:1) to
afford the desired product (220 mg, 18% yield). ESI-MS m/z: 472.3
[M+H].sup.+.
1-(1-Acryloylazetidin-3-yl)-N-(4,5-dichloro-2-methoxybenzyl)piperidine-4-c-
arboxamide
[0843] A mixture of tert-butyl
3-(4-(4,5-dichloro-2-methoxybenzylcarbamoyl)piperidin-1-yl)azetidine-1-ca-
rboxylate (210 mg, 0.44 mmol) in HCl/MeOH (10 mL, 2.86 M) was
stirred at room temperature for 1 h. The mixture was concentrated
in vacuo to yield the crude residue. The residue was dissolved in
DCM (5 mL), triethylamine (0.5 mL) and acryloyl chloride (40 mg,
0.43 mmol) were added. The reaction mixture was stirred at room
temperature for 30 min and then partitioned between DCM and water.
The organic layer was washed with water and brine, dried over
Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was
purified by flash column chromatography on silica gel
(dichloromethane/methanol=50:1) to afford the desired product (150
mg, 82% yield).
N-(4,5-Dichloro-2-hydroxybenzyl)-1-(1-acryloylazetidin-3-yl)piperidine-4-c-
arboxamide
[0844] To a solution of
1-(1-acryloylazetidin-3-yl)-N-(4,5-dichloro-2-methoxybenzyl)piperidine-4--
carboxamide (150 mg, 0.35 mmol) in DCM (15 mL) at -60.degree. C.,
BBr.sub.3 (0.6 g, 2.4 mmol) was added dropwise. The resulting
mixture was allowed to warm to room temperature and stirred for 1
h. The mixture was poured into ice-water, basified with saturated
NaHCO.sub.3 solution to adjust the pH to 8-9, and then extracted
with DCM. The organic layer was dried over anhydrous sodium sulfate
and concentrated in vacuo. The residue was purified by flash column
chromatography on silica gel (dichloromethane/methanol=20:1) to
afford the desired product (34 mg, 24% yield). .sup.1H NMR (400
MHz, DMSO-d6) .delta.: 10.34 (s, 1H), 8.2-8.25 (m, 1H), 7.2 (s,
1H), 7.0 (s, 1H), 6.91 (s, 1H), 6.33-6.27 (m, 1H), 6.12-6.07 (dd,
J=2.4, 12.4 Hz, 1H), 5.68-5.65 (dd, J=2.4, 10.4 Hz, 1H), 4.24-4.20
(m, 1H), 4.17-4.14 (m, 2H), 4.14-3.99 (m, 1H), 3.94-3.90 (m, 1H),
3.73-3.70 (m, 1H), 3.10 (s, 1H), 2.84-2.80 (m, 2H), 2.22 (m, 1H),
1.80 (s, 2H), 1.73-1.71 (m, 2H), 1.63-1.57 (m, 2H). ESI-MS m/z:
412.2 [M+H].sup.+.
Example 44
Synthesis of
1-(3-(hydroxymethyl)-4-(2',5',6-trichloro-4-methoxybiphenylcarbonyl)piper-
azin-1-yl)prop-2-en-1-one (III-7)
##STR00661##
[0845] Methyl 4-chloro-5-iodo-2-methoxybenzoate
[0846] A mixture of 4-chloro-5-iodo-2-methoxybenzoic acid (2 g,
6.41 mmol) concentrated sulfuric acid (1.5 mL) in MeOH (50 mL) was
stirred at reflux for 16 h. The mixture was allowed to cool to room
temperature and partitioned between water and ethyl acetate. The
organic layer was washed with water and brine, dried over
Na.sub.2SO.sub.4 and concentrated in vacuo to afford the desired
product (1.85 g, 85% yield) as a yellow oil.
Methyl
2',5',6-trichloro-4-methoxy-[1,1'-biphenyl]-3-carboxylate
[0847] A mixture of Methyl 4-chloro-5-iodo-2-methoxybenzoate (1.8
g, 5.51 mmol), (2,5-dichlorophenyl)boronic acid (2.1 g, 11.03
mmol), Pd(PPh.sub.3).sub.4(403 mg, 0.55 mmol), Na.sub.2CO.sub.3
(1.75 g, 16.54 mmol) in 1,4-dioxane (50 mL) and water (5 mL) was
stirred at reflux under argon for 16 h. The mixture was filtered
and filtrate was concentrated in vacuo. The residue was purified by
flash column chromatography on silica gel (petroleum ether/ethyl
acetate=20:1) to afford the desired product (1.6 g, 85% yield).
1-(3-(Hydroxymethyl)-4-(2',5',6-trichloro-4-methoxy-[1,1'-biphenyl]-3-carb-
onyl)piperazin-1-yl)prop-2-en-1-one (III-7)
[0848] The title compound was prepared from methyl
2',5',6-trichloro-4-methoxy-[1,1'-biphenyl]-3-carboxylate in three
steps followed the procedure described in Example 36.
tert-butyl3-(hydroxymethyl)-4-(2',5',6-trichloro-4-methoxy-[1,1'-biphenyl-
]-3-carbonyl)piperazine-1-carboxylate (420 mg, 0.79 mmol) was
stirred in HCl in MeOH (2.85 N). The solvent was removed under
reduced pressure to yield the crude reside which was dissolved in
DMF (20 mL), acrylic acid (57 mg, 0.79 mmol), BOP (421 mg, 0.95
mmol) and DIEA (409 mg, 3.17 mmol) were added. The reaction was
stirred at room temperature for 1 h. The resulting mixture was
poured into water, extracted with ethyl acetate and washed with
water and brine. The combined organic layer was dried over
Na.sub.2SO.sub.4, filtered and concentrated. The residue was
purified by column chromatography on silica gel
(dichloromethane/methanol=60:1) to afford the desired product (92
mg, 24% yield). .sup.1H NMR (400 MHz, DMSO-d6) .delta.: 7.64-7.20
(m, 5H), 6.83-6.70 (m, 1H), 6.16-6.11 (d, 1H), 5.74-5.71 (d, 1H),
6.91 (s, 1H), 5.08-4.01 (m, 3H), 3.90-3.86 (d, 3H), 3.49-3.22 (m,
2H), 2.93-2.74 (m, 2H), 2.89-2.67 (m, 2H). ESI-MS m/z: 451.2
[M-H].sup.-.
Example 45
Synthesis of
1-(4-(4-(2,4-dichlorophenyl)-1h-pyrrole-2-carbonyl)piperazin-1-yl)prop-2--
en-1-one (III-32)
##STR00662##
[0849] tert-Butyl
4-(4-bromo-1H-pyrrole-2-carbonyl)piperazine-1-carboxylate
[0850] To a mixture of 4-bromo-1H-pyrrole-2-carboxylic acid (800
mg, 4.21 mmol), tert-butylpiperazine-1-carboxylate (822 mg, 4.42
mmol), BOP (2.2 g, 5.05 mmol) in DMF (5 mL), DIEA (1.63 g, 12.63
mmol) was added and the resulting mixture was stirred at room
temperature for 1 h. The mixture was poured into water and
extracted with ethyl acetate. The organic layer was washed with
water and brine, dried over Na.sub.2SO.sub.4 and concentrated in
vacuo to afford the crude product (920 mg, 61% yield) which was
used directly in the next step without purification.
tert-Butyl
4-(4-(2,4-dichlorophenyl)-1H-pyrrole-2-carbonyl)piperazine-1-ca-
rboxylate
[0851] A mixture of tert-butyl
4-(4-bromo-1H-pyrrole-2-carbonyl)piperazine-1-carboxylate (350 mg,
0.98 mmol), (2,4-dichlorophenyl) boronic acid (280 mg, 1.47 mmol),
Pd(PPh.sub.3).sub.4(116 mg, 0.1 mmol), Na.sub.2CO.sub.3(312 mg,
2.94 mmol) in 1,4-dioxane (10 mL) and water (2 mL) was stirred at
reflux under argon for 16 h. The mixture was allowed to cool to
room temperature and concentrated in vacuo. The residue was
purified by flash column chromatography on silica gel (petroleum
ether/ethyl acetate=30:1) to afford the desired product (273 mg,
59% yield).
1-(4-(4-(2,4-Dichlorophenyl)-1H-pyrrole-2-carbonyl)piperazin-1-yl)prop-2-e-
n-1-one (VI-32)
[0852] The mixture of tert-butyl
4-(4-(2,4-dichlorophenyl)-1H-pyrrole-2-carbonyl)piperazine-1-carboxylate
(270 mg, 0.64 mmol) in HCl/MeOH (20 mL, 57.2 mmol) was stirred for
1 h. The mixture was concentrated in vacuo and the residue was
dissolved in DMF (5 mL). To this mixture, acrylic acid (50 mg, 0.7
mmol), BOP (437 mg, 0.72 mmol) and DIEA (248 mg, 1.92 mmol) were
added. The reaction mixture was stirred at room temperature for 1
h. The mixture was partitioned between ethyl acetate and water. The
organic layer was washed with brine, dried over Na.sub.2SO.sub.4
and concentrated in vacuo. The residue was purified by flash column
chromatography on silica gel (dichloromethane/methanol=60:1) to
afford the desired product (40 mg, 27% yield) as a white solid.
.sup.1H NMR (400 MHz, DMSO-d6) .delta.: 11.88 (s, 1H), 7.66-7.63
(m, 2H), 7.43 (m, 1H), 7.38 (s, 1H), 6.91 (s, 1H), 6.85-6.78 (m,
1H), 6.18-6.13 (dd, J=2.4, 12.4 Hz, 1H), 5.74-5.71 (dd, J=2.4, 10.4
Hz, 1H), 3.76 (s, 4H), 3.68-3.63 (m, 4H). ESI-MS m/z: 377.3
[M-H].sup.-.
Example 46
Synthesis of
(e)-1-(4-(2',6-dichloro-4-hydroxybiphenylcarbonyl)piperazin-1-yl)-4-(dime-
thylamino)but-2-en-1-one (III-24)
##STR00663##
[0853] tert-Butyl
4-(2',6-dichloro-4-methoxy-[1,1'-biphenyl]-3-carbonyl)piperazine-1-carbox-
ylate
[0854] To a stirred solution of
2',6-dichloro-4-methoxy-[1,1'-biphenyl]-3-carboxylic acid (500 mg,
1.68 mmol) in DMF (10 mL) at room temperature, tert-butyl
piperazine-1-carboxylate (345 mg, 1.85 mmol), BOP (892 mg, 2.02
mmol) and DIEA (542 mg, 4.2 mmol) were added and the resulting
mixture was stirred at room temperature for 2 h. The reaction
mixture was diluted with ethyl acetate and washed with brine. The
organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered
and concentrated in vacuo. The residue was purified by flash column
chromatography on silica gel (dichloromethane/methanol=50:1) to
afford the desired product (550 mg, 70% yield). ESI-MS m/z: 465.4
[M+H].sup.+.
(E)-1-(4-(2',6-dichloro-4-methoxy-[1,1'-biphenyl]-3-carbonyl)piperazin-1-y-
l)-4-(dimethylamino)but-2-en-1-one
[0855] A mixture of tert-Butyl
4-(2',6-dichloro-4-methoxy-[1,1'-biphenyl]-3-carbonyl)piperazine-1-carbox-
ylate (550 mg, 1.18 mmol) in HCl/MeOH (20 mL, 57.2 mmol) was
stirred at room temperature for 1 h. The solvent was removed under
reduced pressure to yield the crude product. The crude residue was
dissolved with DMF (10 mL), 4-(dimethylamino)but-2-enoic acid (215
mg, 0.47 mmol), BOP (627 mg, 1.42 mmol) and DIEA (610 mg, 4.73
mmol) was added. The resulting mixture was stirred at room
temperature for 2 h. The reaction mixture was diluted with ethyl
acetate and washed with brine. The organic layer was dried over
anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The
residue was purified by flash column chromatography on silica gel
(dichloromethane/methanol=50:1) to afford the desired product (450
mg, 80% yield, 2 steps). ESI-MS m/z: 476.4 [M+H].sup.+.
(E)-1-(4-(2',6-dichloro-4-hydroxy-[1,1'-biphenyl]-3-carbonyl)piperazin-1-y-
l)-4-(dimethylamino)but-2-en-1-one (VI-24)
[0856] A solution of
(E)-1-(4-(2',6-dichloro-4-methoxy-[1,1'-biphenyl]-3-carbonyl)piperazin-1--
yl)-4-(dimethylamino)but-2-en-1-one (270 mg, 0.57 mmol) in DCM (5
mL) at -78.degree. C., BBr.sub.3 (1.43 g, 5.7 mmol) was added
dropwise. The resulting mixture was stirred at room temperature for
2 h. The mixture was poured into ice-water, basified with the
aqueous NaHCO.sub.3 to adjust the pH to 7 and then extracted with
DCM (3.times.20 mL). The organic layer was dried over
Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was
purified by column chromatography on silica gel
(dichloromethane/methanol=50:1) to afford the desired product (150
mg, 57% yield). .sup.1HNMR (400 MHz, DMSO-d6) .delta.: 13.12 (s,
1H), 8.02 (s, 1H), 7.81 (s, 1H), 6.81 (m, 1H), 6.13 (dd, J=2.8,
16.8 Hz, 1H), 5.71 (dd, J=2.0, 10.4 Hz, 1H), 4.10 (s, 2H),
3.50-3.60 (m, 8H). ESI-MS m/z: 462.4 [M+H].sup.+.
Example 47
Synthesis of
1-(4-(2',6-dichloro-4-methoxybiphenylcarbonyl)piperazin-1-yl)prop-2-en-1--
one (III-1)
##STR00664##
[0857] Example 48
Synthesis of
1-(3-(4-(2-(4-chloro-5-cyclobutyl-2-hydroxyphenylamino)acetyl)piperazin-1-
-yl)azetidin-1-yl)prop-2-en-1-one (II-53)
##STR00665##
[0858] tert-Butyl
3-(4-(2-(4-Chloro-5-iodo-2-methoxyphenylamino)acetyl)piperazin-1-yl)azeti-
dine-1-carboxylate
[0859] To a solution of
2-(4-chloro-5-iodo-2-methoxyphenylamino)acetic acid (2.0 g, 5.88
mmol), tert-butyl 3-(piperazin-1-yl)azetidine-1-carboxylate (1.84
g, 7.64 mmol), EDCI.HCl (2.26 g, 11.76 mmol), and HOBt (1.59 g,
11.76 mmol) in DMF (3 mL) at 0.degree. C., Et.sub.3N (3.28 mL,
23.52 mmol) was added. The resulting mixture was stirred at RT for
16 h and then partitioned between ethyl acetate and water. The
organic layer was washed with brine, dried over Na.sub.2SO.sub.4
and concentrated in vacuo. The residue was washed by a mixture of
ethyl acetate/petroleum ether=1:5 to afford the desired product
(2.24 g, 67% yield) as a white solid. ESI-MS m z: 565.4
[M+H].sup.+.
tert-Butyl
3-(4-(2-(4-chloro-5-cyclobutyl-2-methoxyphenylamino)acetyl)pipe-
razin-1-yl)azetidine-1-carboxylate
[0860] A mixture of tert-butyl
3-(4-(2-(4-chloro-5-iodo-2-methoxyphenylamino)acetyl)piperazin-1-yl)azeti-
dine-1-carboxylate (697 mg, 1.24 mmol), cyclobutylzinc bromide
(4.46 mL, 2.23 mmol, 0.5 M in THF), Pd(OAc).sub.2 (56 mg, 0.248
mmol), and S-Phos (102 mg, 0.248 mmol) in THE (15 mL) was stirred
at 65.degree. C. under argon for 16 h. The mixture was allowed to
cool to RT, quenched with aqueous NH.sub.4Cl solution and then
extracted with ethyl acetate. The organic layer was washed with
brine, dried over Na.sub.2SO.sub.4, filtered and concentrated in
vacuo. The residue was purified by flash column chromatography on
silica gel (methanol/dichloromethane=1:30) to afford the desired
product (596 mg, 98% yield) as a brown oil. ESI-MS m/z: 493.5
[M+H].sup.+.
1-(3-(4-(2-(4-Chloro-5-cyclobutyl-2-hydroxyphenylamino)acetyl)piperazin-1--
yl)azetidin-1-yl)prop-2-en-1-one (V-53)
[0861] The title compound was prepared from tert-butyl
3-(4-(2-(4-chloro-5-cyclobutyl-2-methoxyphenylamino)acetyl)piperazin-1-yl-
)azetidine-1-carboxylate in three steps according to the procedure
described in Example 33. .sup.1H NMR (400 MHz, DMSO-d6) .delta.:
9.66 (s, 1H), 6.63 (s, 1H), 6.52 (s, 1H), 6.31 (dd, J=10.2, 16.9
Hz, 1H), 6.10 (dd, J=2.1, 16.8 Hz, 1H), 5.68 (dd, J=2.1, 10.2 Hz,
1H), 5.16 (t, J=4.4 Hz, 1H), 4.27-4.23 (m, 1H), 4.08-4.04 (m, 1H),
3.97-3.93 (m, 3H), 3.80-3.76 (m, 1H), 3.65-3.59 (m, 1H), 3.56-3.54
(m, 4H), 3.20-3.14 (m, 1H), 2.40-2.25 (m, 4H), 2.20-2.15 (m, 2H),
2.09-2.05 (m, 2H), 1.97-1.90 (m, 1H), 1.80-1.74 (m, 1H). ESI-MS
m/z: 433.4 [M+H].sup.+.
Example 49
Synthesis of
1-(3-(4-(2-(4-chloro-5-cyclobutyl-2-hydroxyphenylamino)propanoyl)piperazi-
n-1-yl)azetidin-1-yl)prop-2-en-1-one (II-59)
##STR00666##
[0862] Methyl
2-(4-chloro-5-iodo-2-methoxyphenylamino)propanoate
[0863] A mixture of tert-butyl 4-chloro-5-iodo-2-methoxybenzenamine
(2 g, 7.07 mmol), methyl 2-bromopropanoate (1.17 g, 7.07 mmol),
K.sub.2CO.sub.3 (1.94 g, 14.14 mmol) and KI (0.235 g, 1.414 mmol)
in DMF (25 mL) was stirred at 100.degree. C. for 16 h. The mixture
was allowed to cool to RT, quenched with aqueous NaHCO.sub.3
solution and then extracted with ethyl acetate. The organic layer
was washed with brine, dried over Na.sub.2SO.sub.4, filtered and
concentrated in vacuo. The residue was purified by flash column
chromatography on silica gel (ethyl acetate/petroleum ether=1:20)
to afford the desired product (1.12 g, 43% yield) as a yellow
solid. ESI-MS m/z: 370.1 [M+H].sup.+.
2-(4-Chloro-5-iodo-2-methoxyphenylamino)propanoic acid
[0864] To a solution of methyl
2-(4-chloro-5-iodo-2-methoxyphenylamino)propanoate (1.12 g, 3.04
mmol) in mixture of tetrahydrofuran (20 mL) and water (10 mL) at
RT, LiOH.H.sub.2O (0.51 g, 12.16 mmol) was added and the resulting
mixture was stirred for 1 h. The aqueous phase was washed with TBME
and then acidified with aqueous HCl (1 N) to adjust the pH to 5.
The mixture was extracted with ethyl acetate. The organic layer was
washed with brine, dried over anhydrous Na.sub.2SO.sub.4, filtered
and concentrated in vacuo to afford the crude product (760 mg)
which was used directly in the next step without further
purification. ESI-MS m/z: 356.1 [M+H].sup.+.
Tert-Butyl
3-(4-(2-((4-Chloro-5-iodo-2-methoxyphenyl)amino)propanoyl)piper-
azin-1-yl)azetidine-1-carboxylate
[0865] To a solution of
2-(4-chloro-5-iodo-2-methoxyphenylamino)propanoic acid (760 mg,
2.13 mmol), tert-butyl 3-(piperazin-1-yl)azetidine-1-carboxylate
(669 mg, 2.78 mmol), EDCI.HCl (818 mg, 4.26 mmol), HOBt (575 mg,
4.26 mmol) in DMF (8 mL) at 0.degree. C., Et.sub.3N (861 mg, 8.52
mmol) was added. The resulting mixture was stirred at RT for 16 h
and then partitioned between ethyl acetate and water. The organic
layer was washed with saturated NaHCO.sub.3 solution and brine,
dried over Na.sub.2SO.sub.4 and concentrated in vacuo. The residue
was purified by flash column chromatography on silica gel (ethyl
acetate/petroleum ether=1:1) to afford the desired product (673 mg,
55% yield) as a white solid. ESI-MS m/z: 579.4 [M+H].sup.+.
Tert-Butyl
3-(4-(2-((4-Chloro-5-cyclobutyl-2-methoxyphenyl)amino)propanoyl-
)piperazin-1-yl)azetidine-1-carboxylate
[0866] A mixture of tert-butyl
3-(4-(2-((4-chloro-5-iodo-2-methoxyphenyl)amino)propanoyl)piperazin-1-yl)-
azetidine-1-carboxylate (673 mg, 1.162 mmol), cyclobutylzinc
bromide (5.11 mL, 2.556 mmol, 0.5 M in THF), Pd(Oac).sub.2 (52 mg,
0.23 mmol), S-Phos (95 mg, 0.23 mmol) in THF (10 mL) was stirred at
65.degree. C. under argon for 16 h. The mixture was allowed to cool
to RT, quenched with aqueous NH.sub.4Cl solution and then extracted
with ethyl acetate. The organic layer was washed with brine, dried
over Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The
residue was purified by flash column chromatography on silica gel
(ethyl acetate/petroleum ether=1:1) to afford the desired product
(565 mg, 96% yield) as a light yellow solid. ESI-MS m/z: 507.6
[M+H].sup.+.
1-(3-(4-(2-((4-Chloro-5-cyclobutyl-2-hydroxyphenyl)amino)propanoyl)piperaz-
in-1-yl)azetidin-1-yl)prop-2-en-1-one (V-59)
[0867] The title compound was prepared from tert-butyl
3-(4-(2-((4-chloro-5-cyclobutyl-2-methoxyphenyl)amino)propanoyl)piperazin-
-1-yl)azetidine-1-carboxylate in three steps according to the
procedure described in Example 33. .sup.1H NMR (400 MHz, DMSO-d6)
.delta.: 9.63 (s, 1H), 6.62 (s, 1H), 6.49 (s, 1H), 6.30 (dd,
J=10.1, 16.8 Hz, 1H), 6.10 (d, J=18.7 Hz, 1H), 5.68 (d, J=10.4 Hz,
1H), 4.86 (d, J=9.2 Hz, 1H), 4.69-4.63 (m, 1H), 4.27-4.23 (m, 1H),
4.07-4.03 (m, 1H), 3.97-3.62 (m, 1H), 3.82-3.76 (m, 2H), 3.64-3.55
(m, 3H), 3.77-3.11 (m, 1H), 2.44-2.15 (m, 6H), 2.08-1.90 (m, 4H),
1.80-1.72 (m, 2H), 1.97-1.90 (m, 1H), 1.24 (d, J=6.4 Hz, 3H).
ESI-MS m/z: 447.4 [M+H].sup.+.
Example 50
Synthesis of tert-butyl
4-(1-acryloylazetidin-3-yl)piperazine-1-carboxylate
##STR00667##
[0868] 1-Benzhydrylazetidin-3-yl methanesulfonate
[0869] A mixture of 1-benzhydrylazetidin-3-ol (20.0 g, 83.68 mmol)
and Et.sub.3N (12.68 g, 125.52 mmol) in DCM (200 mL) at 0.degree.
C., MsCl (11.447 mg, 100.41 mmol) was added in portions and the
resulting solution was stirred at RT for 1 h. The reaction mixture
was diluted with ethyl acetate and washed with brine. The organic
layer was dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated in vacuo to afford the desired product (26.526 g, 100%
yield).
tert-Butyl
4-(1-benzhydrylazetidin-3-yl)piperazine-1-carboxylate
[0870] A mixture of 1-benzhydrylazetidin-3-yl methanesulfonate
(26.53 g, 83.68 mmol), tert-butyl piperazine-1-carboxylate (18.68
g, 100.41 mmol) and K.sub.2CO.sub.3 (23.09 g, 163.36 mmol) in
CH.sub.3CN (200 mL) was stirred at 80.degree. C. for 16 h. The
reaction mixture was cooled to RT and diluted with ethyl acetate
and washed with brine. The organic layer was dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The residue
was purified by flash column chromatography on silica gel
(dichloromethane/methanol=50:1) to afford the desired product (25.5
g, 80% yield).
tert-Butyl 4-(azetidin-3-yl)piperazine-1-carboxylate
[0871] A mixture of tert-butyl
4-(1-benzhydrylazetidin-3-yl)piperazine-1-carboxylate (10.0 g,
24.57 mmol) and 10% Pd\C (2.5 g) in MeOH (100 mL) was stirred under
H.sub.2 atmosphere at 50.degree. C. for 48 h. The reaction mixture
was cooled to RT and filtered. The filtrate was diluted
concentrated in vacuo to afford a crude desire product (6.7 g) as a
colorless oil.
tert-Butyl 4-(1-acryloylazetidin-3-yl)piperazine-1-carboxylate
[0872] A mixture of tert-butyl
4-(azetidin-3-yl)piperazine-1-carboxylate (6.7 g, 27.80 mmol) and
Et.sub.3N (8.43 g, 83.40 mmol) in DCM (100 mL) at 0.degree. C.,
acryloyl chloride (3.77 g, 41.7 mmol) was added in portions and the
resulting solution was stirred at RT for 1 h. The reaction mixture
was diluted with DCM and washed with brine. The organic layer was
dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in
vacuo. The residue was purified by column chromatography on silica
gel (dichloromethane/methanol=50:1) to afford the desired product
(3.6 g, 49.66% yield, 2 steps). .sup.1H NMR (400 MHz, DMSO-d6)
.delta.: 6.30 (dd, J=10.4, 16.8 Hz, 1H), 6.09 (dd, J=2.4, 17.2 Hz,
1H), 5.67 (dd, J=2.4, 10.4 Hz, 1H), 4.22 (t, J=8, 1H), 4.03-4.00
(m, 1H), 3.94-3.90 (m, 1H), 3.75-3.70 (m, 1H), 3.32 (t, J=8.8 Hz,
4H), 3.10-3.18 (m, 1H), 2.22-2.30 (m, 1H), 1.40 (s, 9H).
Example 51
Synthesis of
1-acryloyl-4-(4',6-dichloro-4-hydroxybiphenylcarbonyl)piperazine-2-carbon-
itrile (iII-37)
##STR00668##
[0873] Piperazine-2-carbonitrile
[0874] To a mixture of tert-butyl 3-cyanopiperazine-1-carboxylate
(200 mg, 0.95 mmol) in dichloromethane (10 mL), CF.sub.3COOH (2 mL)
was added and the resulting was stirred at RT for 1 h. The mixture
was concentrated in vacuo to afford the crude product.
4-(4',6-Dichloro-4-methoxy-[1,1'-biphenyl]-3-carbonyl)piperazine-2-carboni-
trile
[0875] To a mixture of
4',6-dichloro-4-methoxy-[1,1'-biphenyl]-3-carboxylic acid (309 mg,
1.04 mmol), EDCI (272 mg, 1.43 mmol), HOBt (195 mg, 1.43 mmol),
Et.sub.3N (288 mg, 2.85 mmol) in dichloromethane (10 mL) at
0.degree. C., piperazine-2-carbonitrile was added at 0.degree. C.
and the resulting mixture was stirred at RT for 8 h. The mixture
was partitioned between dichloromethane and water. The organic
layer was washed brine, dried over MgSO.sub.4, filtered and
concentrated in vacuo. The residue was purified by flash column
chromatography on silica gel (dichloromethane/methanol=50:1) to
afford the desired product (225 mg, 61% yield). ESI-MS m/z: 444.3
[M+H].sup.+.
1-Acryloyl-4-(4',6-dichloro-4-hydroxy-[1,1'-biphenyl]-3-carbonyl)piperazin-
e-2-carbonitrile (III-37)
[0876] The title compound was prepared from
4-(4',6-dichloro-4-methoxy-[1,1'-biphenyl]-3-carbonyl)piperazine-2-carbon-
itrile in two steps according to the procedure described in Example
33. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.: 9.24 (s, 1H),
7.44-7.33 (m, 5H), 7.20 (s, 1H), 6.57-6.45 (m, 2H), 6.79 (s, 1H),
5.94-5.91 (m, 1H), 5.75 (s, 1H), 4.62-4.61 (m, 1H), 4.50-4.46 (m,
1H), 4.06 (s, 1H), 3.61 (s, 1H), 3.36-3.33 (m, 1H), 3.16-3.10 (m,
1H). ESI-MS m/z: 428.4 [M+H].sup.+.
Example 52
Synthesis of
1-(3-(4-(3-(4-chloro-5-cyclopropyl-2-hydroxyphenyl)propanoyl)piperazin-1--
yl)azetidin-1-yl)prop-2-en-1-one (II-58)
##STR00669##
[0877] 5-Bromo-4-chloro-2-methoxybenzenamine
[0878] To a solution of 2,4-dichloro-1-nitrobenzene (100 g, 0.52
mol) in DMSO (200 mL), aqueous solution of NaOH (41.6 g, 1.04 mol)
in water (42 mL) was added and the resulting mixture was stirred
60.degree. C. for 16 h. The mixture was allowed to cool to room
temperature, poured to ice water, and then acidified with aqueous
HCl (1 M) to adjusted the pH to 3-4. The mixture was extracted with
ethyl acetate. The combined organic layer was washed with brine,
dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuo.
The residue was used directly in the next step (80 g, 88%
yield).
[0879] To a solution of 5-chloro-2-nitrophenol (40 g, 0.23 mol) in
DMF (200 mL), K.sub.2CO.sub.3 (47.6 g, 0.345 mol) and iodomethane
(49 g, 0.345 mol) were added and the resulting mixture was stirred
at room temperature for 16h. The mixture was partitioned between
ethyl acetate and water. The organic layer was washed with brine,
dried over MgSO.sub.4, filtered and concentrated in vacuo. The
residue was purified by flash column chromatography on silica gel
(petroleum ether) to afford the desired product (30 g, 70%
yield).
[0880] To a solution of H.sub.2SO.sub.4 (600 mL, 90%),
trifluoromethanesulfonic anhydride (11.3 g, 0.04 mol) and NIS
(49.68 g, 0.22 mol) were added and resulting mixture was stirred at
room temperature for 1 h. To this mixture,
4-chloro-2-methoxy-1-nitrobenzene (69 g, 0.368 mol) was added
quickly. The mixture was stirred for 1 h, and then NIS (33.12 g,
0.148 mol) was slowly added to the mixture. The mixture was stirred
at room temperature for 1 h and then was poured into ice-water. The
precipitate collected by filtration, rinsed with water, aqueous
NaSO.sub.3 and NaHCO.sub.3 solutions, and then dried in vacuo to
afford the desired product (113 g, 98% yield).
[0881] To a solution of 1-chloro-2-iodo-5-methoxy-4-nitrobenzene
(113 g, 0.361 mol) in acetic acid (1 L) and water (50 mL) at
50.degree. C., Fe (50.5 g, 0.903 mol) was added and the resulting
mixture was stirred at 50.degree. C. for 2 h. The mixture was
allowed to cool to room temperature and then poured into ice-water.
The precipitate was collected by filtration and rinsed with water.
This crude product was dissolved with ethyl acetate (1 L) and
filtered.
[0882] The filtrate was washed with saturated NaHCO.sub.3 solution
and brine. The organic layer was dried over MgSO.sub.4, filtered,
and concentrated in vacuo to afford the desired product (87 g, 85%
yield).
1-Bromo-2-chloro-5-iodo-4-methoxybenzene
[0883] To a mixture of 5-bromo-4-chloro-2-methoxyaniline (3 g, 12.7
mmol) in 6N HCl (60 mL, 360 mmol) at 0.degree. C., a solution of
NaNO.sub.2 (963 mg, 13.9 mmol) in water (20 mL) was added dropwise
while keeping the internal temperature around 0.degree. C. KI (10.5
g, 63.4 mmol) and CuI (4.8 g, 25.4 mmol) were dissolved in water
(20 mL) and added to the stirred reaction mixture. The reaction was
kept at 5.degree. C. for 2 h. The reaction mixture was extracted
with ethyl acetate. The combined organic layer was washed with
water, Na.sub.2SO.sub.3 (aq, 10%) and brine, dried over anhydrous
Na.sub.2SO.sub.4, an concentrated. The residue was purified by
flash column chromatography on silica gel (ethyl acetate/petroleum
ether=1:100) to afford the desired product (3.2 g, 73% yield).
3-(5-Bromo-4-chloro-2-methoxyphenyl)propanal
[0884] A mixture of 1-bromo-2-chloro-5-iodo-4-methoxybenzene (3.2
g, 9.2 mmol), prop-2-en-1-ol (1.3 g, 23.0 mmol), Pd(OAc).sub.2 (206
mg, 0.9 mmol), TBAC (2.56 g, 9.2 mmol), NaHCO.sub.3 (2.3 g, 27.6
mmol) in DMF (50 mL) was stirred under Argon at 60.degree. C. for
16 h. The mixture was allowed to cool to RT, and then partitioned
between ethyl acetate and water. The combined organic layer was
washed with brine, dried over anhydrous Na.sub.2SO.sub.4 and
concentrated. The residue was purified by flash column
chromatography on silica gel (ethyl acetate/petroleum ether=1:20)
to afford the desired product (860 mg, 34% yield).
3-(5-Bromo-4-chloro-2-methoxyphenyl)propanoic acid
[0885] To a stirred solution of Jones reagent (3 mL, 5.4 mmol, 2.8
M) in acetone (20 mL), 3-(5-bromo-4-chloro-2-methoxyphenyl)propanal
(860 mg, 3.1 mmol) was added. The reaction was stirred at RT for 12
h, quenched with iso-propylalcohol and then stirred for 10 min. The
resulting mixture was diluted with water, extracted with ethyl
acetate. The combined organic layer was washed with water and
brine, dried over Na.sub.2SO.sub.4 and concentrated. The residue
was purified by flash column chromatography on silica gel (ethyl
acetate/petroleum ether=1:1) to afford the desired product (358 mg,
38% yield). ESI-MS m/z: 291.1 [M+H].sup.-.
tert-Butyl-3-(4-(3-(5-bromo-4-chloro-2-methoxyphenyl)propanoyl)piperazin-1-
-yl)azetidine-1-carboxylate
[0886] To a stirred solution of
3-(5-bromo-4-chloro-2-methoxyphenyl)propanoic acid (350 mg, 1.2
mmol) in DMF (30 mL) at RT, tert-butyl
3-(piperazin-1-yl)azetidine-1-carboxylate (317 mg, 1.3 mmol), BOP
(731 mg, 1.4 mmol) and DIEA (461 mg, 3.6 mmol) were added and the
mixture was stirred at RT for 1 h. The reaction mixture was
partitioned between ethyl acetate and water. The organic layer was
washed with brine and dried over anhydrous Na.sub.2SO.sub.4,
filtered and concentrated in vacuo. The residue was purified by
flash column chromatography on silica gel
(dichloromethane/methanol=50:1) to afford the desired product (285
mg, 46% yield).
tert-Butyl
3-(4-(3-(4-chloro-5-cyclopropyl-2-hydroxyphenyl)propanoyl)piper-
azin-1-yl)azetidine-1-carboxylate
[0887] A mixture of tert-butyl
3-(4-(3-(5-bromo-4-chloro-2-methoxyphenyl)propanoyl)piperazin-1-yl)azetid-
ine-1-carboxylate (280 mg, 0.54 mmol), cyclopropylboronic acid (185
mg, 2.2 mmol), K.sub.3PO.sub.4.3H.sub.2O (444 mg, 1.9 mmol),
tricyclohexylphosphine (30 mg, 0.1 mmol), Pd(OAc).sub.2 (24 mg,
0.11 mmol) in toluene (10 mL) and water (1 mL) was stirred at
reflux under argon for 16 h. The mixture was allowed to cool to RT
and concentrated in vacuo. The residue was purified by flash column
chromatography on silica gel (dichloromethane/methanol=60:1) to
afford the desired product (194 mg, 75% yield). ESI-MS m/z: 477.3
[M+H].sup.+.
1-(3-(4-(3-(4-Chloro-5-cyclopropyl-2-hydroxyphenyl)propanoyl)piperazin-1-y-
l)azetidin-1-yl)prop-2-en-1-one (II-58)
[0888] The title compound was prepared from tert-butyl
3-(4-(3-(4-chloro-5-cyclopropyl-2-hydroxyphenyl)propanoyl)piperazin-1-yl)-
azetidine-1-carboxylate in three steps according to the procedure
described in Example 33. .sup.1H NMR (400 MHz, DMSO-d6) .delta.:
9.68 (s, 1H), 6.79 (s, 1H), 6.74 (s, 1H), 6.33-6.26 (m, 1H),
6.12-6.07 (dd, J=1.9, 17.2 Hz, 1H), 5.68-5.65 (dd, J=2.0, 10.2 Hz,
1H), 4.24-4.20 (m, 1H), 4.05-4.01 (m, 1H), 3.94-3.90 (m, 1H),
3.76-3.72 (m, 1H), 3.45-3.42 (m, 4H), 3.13-3.11 (m, 1H), 2.69-2.65
(m, 2H), 2.53-2.51 (m, 2H), 2.25-2.23 (bs, 4H), 1.97-1.93 (m, 1H),
0.90-0.86 (m, 1H), 0.60-0.55 (m, 1H). ESI-MS m/z: 418.4
[M+H].sup.+.
Example 53
Synthesis of
1-(3-(4-(2-(4-chloro-2-hydroxy-5-(1-methylcyclopropyl)phenylamino)acetyl)-
piperazin-1-yl)azetidin-1-yl)prop-2-en-1-one (II-64)
##STR00670##
[0889] Methyl 5-acetamido-2-chloro-4-methoxybenzoate
[0890] To a mixture of methyl 5-amino-2-chloro-4-methoxybenzoate
(3.6 g, 16.7 mmol), Et.sub.3N (6.7 g, 66.8 mmol) and DCM (100 mL)
at RT, acetyl chloride (1.57 g, 20.1 mmol) was added dropwise and
the resulting mixture was stirred for 12h. The reaction mixture was
partitioned between dichloromethane and water. The organic layer
was washed with saturated NaHCO.sub.3 solution and brine, dried
over anhydrous Na.sub.2SO.sub.4 and concentrated in vacuo. The
residue was purified by flash column chromatography on silica gel
(ethyl acetate/petroleum ether=1:1) to afford the desired product
(2.7 g, 63% yield).
N-(4-Chloro-5-(2-hydroxypropan-2-yl)-2-methoxyphenyl)acetamide
[0891] To a solution of methyl
5-acetamido-2-chloro-4-methoxybenzoate (2.7 g, 11.1 mmol), in THF
(40 mL) at -40.degree. C. under Argon, methylmagnesium bromide (21
mL, 21 mmol, 1M in ether) was added dropwise while keeping the
internal temperature at -40.degree. C. Then the mixture was allowed
to warm to RT, and stirred for 2 h. The reaction mixture was poured
into ice-cooled NH.sub.4Cl (10%) solution, and extracted with ethyl
acetate. The combined organic layer was washed with water and
brine, dried over anhydrous Na.sub.2SO.sub.4 and concentrated in
vacuo to afford the desire product (2.3 g, 80% yield).
N-(4-Chloro-2-methoxy-5-(prop-1-en-2-yl)phenyl)acetamide
[0892] To a solution of
N-(4-chloro-5-(2-hydroxypropan-2-yl)-2-methoxyphenyl)acetamide (3.2
g, 12.4 mmol) in DCM (20 mL) at -5.degree. C., SOCl.sub.2 (3.7 g,
37.25 mmol) was added dropwise. The mixture was warmed to RT, and
then stirred at reflux for 2 h. The reaction mixture was
concentrated and the residue was purified by flash column
chromatography on silica gel (ethyl acetate/petroleum ether=3:1) to
afford the desired product (1.9 g, 64% yield).
N-(4-Chloro-2-methoxy-5-(1-methylcyclopropyl)phenyl)acetamide
[0893] To a solution of
N-(4-chloro-2-methoxy-5-(prop-1-en-2-yl)phenyl)acetamide (1.0 g,
4.17 mmol) in toluene (20 mL) at 0.degree. C., CH.sub.2I.sub.2(5.6
g, 20.86 mmol) and Et.sub.2Zn (41.7 mL, 41.7 mmol, 1.0 M in hexane)
was added. The mixture was kept at 0.degree. C. for 30 min, and
then stirred at RT for 16 h. The reaction mixture was quenched with
saturated NH.sub.4Cl solution and stirred for 15 min. The mixture
was concentrated in vacuo to remove toluene and the resulting
mixture was extracted with dichloromethane. The combined organic
layer was washed with water and brine, dried over anhydrous
Na.sub.2SO.sub.4, concentrated to afford the desired product (820
mg, 77% yield).
4-Chloro-2-methoxy-5-(1-methylcyclopropyl)aniline
[0894] A mixture of
N-(4-chloro-2-methoxy-5-(1-methylcyclopropyl)phenyl)acetamide (820
mg, 3.23 mmol), KOH (1.8 g, 32.3 mmol), ethanol (40 mL) and water
(20 mL) was stirred at reflux for 12 h. The reaction mixture was
extracted with ethyl acetate. The combined organic layer was washed
with water and brine, dried over anhydrous Na.sub.2SO.sub.4 and
filtered. The filtrate was concentrated in vacuo and the residue
was purified by flash column chromatography on silica gel (ethyl
acetate/petroleum ether=20:1) to afford the desired product (460
mg, 67% yield). ESI-MS m/z: 212.4 [M+H].sup.+.
Ethyl
2-((4-chloro-2-methoxy-5-(1-methylcyclopropyl)phenyl)amino)acetate
[0895] To a solution of
4-chloro-2-methoxy-5-(1-methylcyclopropyl)aniline (450 mg, 2.13
mmol) in MeOH (20 mL) at RT, AcOH (3 drops) and ethyl glyoxalate
(326 mg, 3.19 mmol, 50% in toluene) were added. The mixture was
stirred at RT for 2 h and then sodium cyanoborohydride (403 mg,
6.39 mmol) was added to the mixture. The resulting mixture was
stirred at 50.degree. C. for 16 h. The mixture was allowed to cool
to RT, and partitioned between ethyl acetate and water. The organic
layer was dried over MgSO.sub.4, filtered, and concentrated in
vacuo to afford the crude product (636 mg). ESI-MS m/z: 298.2
[M+H].sup.+.
2-((4-Chloro-2-methoxy-5-(1-methylcyclopropyl)phenyl)amino)acetic
acid
[0896] To a solution of ethyl
2-((4-chloro-2-methoxy-5-(1-methylcyclopropyl)phenyl)amino)acetate
(630 mg, 2.12 mmol) in THF (15 mL) and water (5 mL), LiOH.H.sub.2O
(889 mg, 21.2 mmol) was added and the resulting mixture was stirred
at RT for 2 h. The mixture was washed with 20% ethyl
acetate/petroleum ether. The aqueous layer was acidified with
aqueous HCl (1 N) to adjust pH to 3-4 and extracted with ethyl
acetate. The organic layer was dried over MgSO.sub.4, filtered and
concentrated in vacuo to afford the desired product (200 mg, 33%
yield).
tert-Butyl
3-(4-(2-((4-chloro-2-methoxy-5-(1-methylcyclopropyl)phenyl)amin-
o)acetyl)piperazin-1-yl)azetidine-1-carboxylate
[0897] To a solution of
2-((4-chloro-2-methoxy-5-(1-methylcyclopropyl)phenyl)amino)acetic
acid (110 mg, 0.41 mmol) and tert-butyl
3-(piperazin-1-yl)azetidine-1-carboxylate (118 mg, 0.49 mmol) in
DMF (15 mL) at RT, BOP (217 mg, 0.49 mmol) and DIEA (159 mg, 1.23
mmol) were added and the resulting mixture was stirred at RT for 1
h. The mixture was partitioned between ethyl acetate and water. The
organic layer was washed with brine, dried over MgSO.sub.4,
filtered and concentrated in vacuo to afford the desired product
(192 mg, 95% yield).
1-(3-(4-(2-((4-Chloro-2-hydroxy-5-(1-methylcyclopropyl)phenyl)amino)acety-
l)piperazin-1-yl)azetidin-1-yl)prop-2-en-1-one (II-64)
[0898] The title compound was prepared from tert-butyl
3-(4-(2-((4-chloro-2-methoxy-5-(1-methylcyclopropyl)phenyl)amino)acetyl)p-
iperazin-1-yl)azetidine-1-carboxylate in three steps according to
the procedure described in Example 33. .sup.1H NMR (400 MHz,
DMSO-d6) .delta.: 9.70 (s, 1H), 6.64 (s, 1H), 6.51 (s, 1H),
6.35-6.28 (m, 1H), 6.13-6.08 (dd, J=1.9, 17.9 Hz, 1H), 5.69-5.66
(dd, J=2.1, 10.1 Hz, 1H), 5.13-5.11 (m, 1H), 4.25-4.23 (m, 1H),
4.08-4.05 (m, 1H), 3.95-3.91 (m, 3H), 3.80-3.76 (m, 1H), 3.53 (bs,
4H), 3.18-3.16 (m, 1H), 2.38-2.31 (m, 4H), 1.26 (s, 3H), 0.72-0.64
(m, 4H). ESI-MS m/z: 434.4 [M+H].sup.+.
Example 54
Synthesis of
1-(4-(2-chloro-5-hydroxybiphenylcarbonyl)piperazin-1-yl)prop-2-en-1-one
(III-42)
##STR00671##
[0899] 5-Chloro-4-iodo-2-methoxybenzoic acid
[0900] To a stirred solution of 4-amino-5-chloro-2-methoxybenzoic
acid (5 g, 24.8 mmol) in water (10 mL) at 0.degree. C.,
concentrated sulfuric acid (50 mL) was added. Then a solution of
NaNO.sub.2 (1.9 g, 27.3 mmol) in water (10 mL) was added dropwise
while keeping the internal temperature around 0.degree. C. KI (4.5
g, 27.3 mmol) and I.sub.2 (3.5 g, 13.64 mmol) were dissolved in
water and added dropwise to the stirred reaction mixture. The
reaction was stirred at 5.degree. C. for 2 h and then extracted
with ethyl acetate. The organic layer was washed with water,
Na.sub.2SO.sub.3 (aq, 10%) and brine, dried over anhydrous
Na.sub.2SO.sub.4 and concentrated in vacuo to afford desired
product (1.55 g, 19% yield). ESI-MS m/z: 311.1 [M+H].sup.+.
tert-Butyl
4-(5-chloro-4-iodo-2-methoxybenzoyl)piperazine-1-carboxylate
[0901] To a stirred solution of 5-chloro-4-iodo-2-methoxybenzoic
acid (1.55 g, 4.9 mmol) in DMF (30 mL) at RT, tert-butyl
piperazine-1-carboxylate (1.02 g, 5.5 mmol), BOP (2.63 g, 25.9
mmol) and DIEA (1.92 g, 14.9 mmol) were added and the resulting
mixture was stirred at RT for 1 h. The reaction mixture was
extracted with ethyl acetate and washed with brine. The organic
layer was dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated in vacuo. The residue was purified by flash column
chromatography on silica gel (dichloromethane/methanol=50:1) to
afford the desired product (1.96 g, 76% yield).
tert-Butyl-4-(2-chloro-5-methoxy-[1,1'-biphenyl]-4-carbonyl)piperazine-1-c-
arboxylate
[0902] A mixture of tert-butyl
4-(5-chloro-4-iodo-2-methoxybenzoyl)piperazine-1-carboxylate (300
mg, 0.56 mmol), phenylboronic acid (82 mg, 0.67 mmol),
Pd(PPh.sub.3).sub.4(129 mg, 0.1 mmol), Na.sub.2CO.sub.3(180 mg,
1.68 mmol) in 1,4-dioxane (10 mL) and water (2 mL) was stirred at
reflux under argon for 16 h. The mixture was allowed to cool to
room temperature and concentrated in vacuo. The residue was
purified by flash column chromatography on silica gel (petroleum
ether/ethyl acetate=30:1) to afford the desired product (219 mg,
80% yield).
1-(4-(2-Chloro-5-hydroxy-[1,1'-biphenyl]-4-carbonyl)piperazin-1-yl)prop-2--
en-1-one (III-42)
[0903] The title compound was prepared from
tert-butyl-4-(2-chloro-5-methoxy-[1,1'-biphenyl]-4-carbonyl)piperazine-1--
carboxylate in three steps according to the procedure described in
Example 33. .sup.1H NMR (400 MHz, DMSO-d6) .delta.: 10.40 (s, 1H),
7.50-7.41 (m, 5H), 7.35 (s, 1H), 6.89 (s, 1H), 6.84 (m, 1H),
6.17-6.13 (d, 1H), 5.73-5.71 (m, 1H), 3.63 (s, 6H), 3.30 (s, 2H).
ESI-MS m/z: 371.2 [M+H].sup.+.
Example 55
Synthesis of
1-(3-(4-(2-(4-chloro-2-hydroxy-5-isopropylphenylamino)acetyl)piperazin-1--
yl)azetidin-1-yl)prop-2-en-1-one (II-50)
##STR00672##
[0904] 4-Chloro-2-methoxy-5-(prop-1-en-2-yl)benzenamine
[0905] A mixture of 4-chloro-5-iodo-2-methoxybenzenamine (1.0 g,
3.53 mmol),
4,4,5,5-tetramethyl-2-(prop-1-en-2-yl)-1,3,2-dioxaborolane (889 mg,
5.29 mmol), Pd(PPh.sub.3).sub.4(363 mg, 0.353 mmol),
Na.sub.2CO.sub.3 (1.12 g, 10.6 mmol) in DME (10 mL) and water (3
mL) was stirred at reflux under argon for 6 h. The reaction mixture
was allowed to cool to RT and extracted with ethyl acetate. The
organic layer was washed with brine, dried over Na.sub.2SO.sub.4
and concentrated in vacuo. The residue was purified by flash column
chromatography on silica gel (5% petroleum ether/ethyl acetate) to
afford the desired product (173 mg, 25% yield) as an off-white
solid. ESI-MS m/z: 198.5[M+H].sup.+.
4-Chloro-5-isopropyl-2-methoxybenzenamine
[0906] A mixture of
4-chloro-2-methoxy-5-(prop-1-en-2-yl)benzenamine (160 mg, 0.81
mmol), Raney-Ni (20 mg) in MeOH (5 mL) was stirred at RT under
H.sub.2 (1 atm) atmosphere for 8 h. The mixture was filtered and
the filtrate was concentrated in vacuo to afford the desired
product (150 mg, 93% yield).
tert-Butyl3-(4-(2-(4-chloro-5-isopropyl-2-methoxyphenylamino)acetyl)pipera-
zin-1-yl)azetidine-1-carboxylate
[0907] The title compound was prepared from
4-chloro-5-isopropyl-2-methoxybenzenamine in three steps according
to the procedure described in Example 45.
1-(3-(4-(2-(4-Chloro-5-isopropyl-2-methoxyphenylamino)acetyl)piperazin-1-y-
l)azetidin-1-yl)prop-2-en-1-one
[0908] A mixture of tert-butyl
3-(4-(2-(4-chloro-5-isopropyl-2-methoxyphenylamino)acetyl)piperazin-1-yl)-
azetidine-1-carboxylate (102 mg, 0.212 mmol) in HCl/MeOH (2.86
.mu.M, 5 mL) was stirred at RT for 1 h. The mixture was
concentrated in vacuo to afford the crude product, the crude was
dissolved in DMF (5 mL) at RT, acrylic acid (17 mg, 0.233 mmol),
BOP (113 mg, 0.254 mmol) and DIEA (82 mg, 0.636 mmol) were added
and the resulting mixture was stirred at RT for 1 h. The mixture
was partitioned between ethyl acetate and water. The organic layer
was washed brine, dried over MgSO.sub.4, filtered and concentrated
in vacuo. The residue was purified by flash column chromatography
on silica gel (dichloromethane/methanol=50:1) to afford the desired
product (77 mg, 85% yield, 2 steps). ESI-MS m/z: 435.4
[M+H].sup.+.
1-(3-(4-(2-(4-Chloro-2-hydroxy-5-isopropylphenylamino)acetyl)piperazin-1-y-
l)azetidin-1-yl)prop-2-en-1-one (II-50)
[0909] To a solution of
1-(3-(4-(2-(4-chloro-5-isopropyl-2-methoxyphenylamino)acetyl)piperazin-1--
yl)azetidin-1-yl)prop-2-en-1-one (77 mg, 0.18 mmol) in DCM (15 mL)
at -60.degree. C., BBr.sub.3 (443 mg, 1.8 mmol) was added dropwise
and the resulting mixture was stirred at RT for 1 h. The mixture
was cooled to -60.degree. C., MeOH was added dropwise and then
basified with Et.sub.3N to adjust the pH to 8-9. The mixture was
poured into water and extracted with dichloromethane. The organic
layer was dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated in vacuo. The residue was purified by flash column
chromatography on silica gel (dichloromethane/methanol=50:1) to
afford the desired product (25 mg, 33% yield). .sup.1H NMR (400
MHz, DMSO-d6) .delta.: 9.50 (bs, 1H), 6.62 (s, 1H), 6.470 (s, 1H),
6.30 (m, 1H), 6.10 (dd, J=2.4, 17.2 Hz, 1H), 5.68 (dd, J=2.0, 10.4
Hz, 1H), 5.14 (m, 1H), 4.25 (m, 1H), 4.06 (m, 1H), 3.96 (m, 1H),
3.91 (m, 2H), 3.78 (m, 1H), 3.54 (m, 4H), 3.17 (m, 2H), 2.35 (m,
4H), 1.21 (m, 6H). ESI-MS m z: 421.4 [M+H].sup.+.
Example 56
Synthesis of
1-(1-acryloylazetidin-3-yl)-4-(2-(4-chloro-5-cyclopropyl-2-hydroxyphenyla-
mino)acetyl)piperazine-2-carboxamide (II-51)
##STR00673##
[0910] 2-Amino-5-chloro-4-cyclopropylphenol
[0911] To a mixture of 2-amino-5-chloro-4-iodophenol (500 mg, 1.9
mmol), PdCl.sub.2(dppf) (136 mg, 0.19 mmol) in THE (10 mL) under
argon at RT, cyclopropylmagnesium bromide (16 mL, 11.4 mmol, 0.7 M
in THF) was added and the mixture was stirred at reflux for 15 h.
The mixture was allowed to cool to RT, and partitioned between
ethyl acetate and water. The organic layer was washed with brine,
dried over Na.sub.2SO.sub.4 and concentrated. The residue was
purified by flash column chromatography on silica gel (10-20% ethyl
acetate/hexanes) to afford the desired product (220 mg, 63% yield)
as a brown solid. .sup.1H NMR (400 MHz, DMSO-d6) .delta.: 9.27 (s,
1H), 6.62 (s, 1H), 6.22 (s, 1H), 4.53 (s, 2H), 1.89-1.93 (m, 1H),
0.83-0.87 (m, 2H), 0.46-0.49 (m, 2H).
Ethyl 2-(4-chloro-5-cyclopropyl-2-hydroxyphenylamino)acetate
[0912] To a solution of 2-amino-5-chloro-4-cyclopropylphenol (200
mg, 1.01 mmol) in MeOH (20 mL) at RT, AcOH (3 drops) and ethyl
glyoxalate (416 mg, 2.02 mmol, 50% in toluene) were added. The
mixture was stirred at RT for 2 h and then sodium cyanoborohydride
(190 mg, 3.03 mmol) was added to the mixture. The resulting mixture
was stirred at 40.degree. C. for 15 h. The mixture was allowed to
cool to RT and partitioned between ethyl acetate and water. The
organic layer was washed with brine, dried over Na.sub.2SO.sub.4
and concentrated. The residue was purified by flash column
chromatography on silica gel (10-20% methanol/dichloromethane) to
afford the desired product (290 mg, 100% yield) as a solid. .sup.1H
NMR (400 MHz, DMSO-d6) .delta.: 9.63 (s, 1H), 6.66 (s, 1H), 5.93
(s, 1H), 5.07 (t, J=6.4 Hz, 1H), 4.12 (q, J=7.2 Hz, 2H), 3.91 (d,
J=6.4 Hz, 2H), 1.92-1.97 (m, 1H), 1.20 (t, J=6.8 Hz, 2H), 0.84-0.87
(m, 2H), 0.51-0.55 (m, 2H).
2-(4-Chloro-5-cyclopropyl-2-hydroxyphenylamino)acetic acid
[0913] To a solution of ethyl
2-(4-chloro-5-cyclopropyl-2-hydroxyphenylamino)acetate (290 mg,
0.89 mmol) in of 4:1 mixture of tetrahydrofuran and water (30 mL)
at RT, LiOH.H.sub.2O (226 mg, 5.34 mmol) was added and the
resulting mixture was stirred for 2 h at 60.degree. C. The mixture
was acidified with aqueous HCl (1 N) to adjust the pH to 3-5 and
then extracted with ethyl acetate. The organic layer was washed
with brine, dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated in vacuo to afford the product (100 mg, 47% yield).
.sup.1H NMR (400 MHz, DMSO-d6) .delta.: 9.64 (s, 1H), 6.66 (s, 1H),
5.96 (s, 1H), 3.81 (s, 2H), 1.89-1.96 (m, 1H), 0.84-0.87 (m, 2H),
0.54-0.56 (m, 2H).
tert-Butyl
4-(1-benzhydrylazetidin-3-yl)-3-carbamoylpiperazine-1-carboxyla-
te
[0914] A mixture of 1-benzhydrylazetidin-3-yl methanesulfonate
(2.69 g, 8.5 mmol), K.sub.2CO.sub.3 (1.76 g, 12.8 mmol), tert-butyl
3-carbamoylpiperazine-1-carboxylate (1.95 g, 8.5 mmol) in
CH.sub.3CN (40 mL) was stirred at reflux for 16 h. The mixture was
partitioned between ethyl acetate and water. The organic layer was
washed with water and brine, dried over Na.sub.2SO.sub.4, and
concentrated in vacuo. The residue was purified by flash column
chromatography on silica gel (dichloromethane/methanol=50:1) to
afford the desired product. (2.08 g, 54% yield).
tert-Butyl
4-(azetidin-3-yl)-3-carbamoylpiperazine-1-carboxylate
[0915] A mixture of
4-chloro-2-methoxy-5-(prop-1-en-2-yl)benzenamine (1 g, 2.22 mmol),
Pd/C (300 mg) in MeOH (25 mL) was stirred at 50.degree. C. under
H.sub.2 (1 atm) atmosphere for 12 h. The mixture was cooled and
filtered. The filtrate was concentrated in vacuo to afford the
desired product (640 mg, 100% yield).
tert-Butyl
4-(1-acryloylazetidin-3-yl)-3-carbamoylpiperazine-1-carboxylate
[0916] To a solution of tert-butyl
4-(azetidin-3-yl)-3-carbamoylpiperazine-1-carboxylate (640 mg, 2.22
mmol) and Et.sub.3N (463 mg, 4.58 mmol) in DCM (10 mL) at 0.degree.
C., acryloyl chloride (248 mg, 2.74 mmol) was added dropwise and
the resulting mixture was stirred at RT for 1.5 h. The mixture was
partitioned between dichloromethane and saturated NaHCO.sub.3
solution. The organic layer was washed with saturated brine, dried
over Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was
purified by flash column chromatography on silica gel
(dichloromethane/methanol=50:1) to afford the desired product (350
mg, 47% yield).
1-(1-acryloylazetidin-3-yl)-4-(2-(4-chloro-5-cyclopropyl-2-hydroxyphenylam-
ino)acetyl)piperazine-2-carboxamide (II-51)
[0917] A mixture of tert-butyl
4-(1-acryloylazetidin-3-yl)-3-carbamoylpiperazine-1-carboxylate
(120 mg, 0.35 mmol) in HCl/MeOH (2.86 .mu.M, 10 mL) was stirred at
RT for 1 h. The mixture was concentrated in vacuo to afford the
crude residue. It was dissolved in DMF (5 mL) at 0.degree. C.,
2-(4-chloro-5-cyclopropyl-2-hydroxyphenylamino)acetic acid (31 mg,
0.427 mmol), BOP (206 mg, 0.466 mmol) and K.sub.2CO.sub.3 (150 mg,
1.164 mmol) were added and the resulting mixture was stirred at RT
for 1 h. The mixture was partitioned between ethyl acetate and
water. The organic layer was washed brine, dried over MgSO.sub.4,
filtered and concentrated in vacuo. The residue was purified by
flash column chromatography on silica gel
(dichloromethane/methanol=20:1) to afford the desired product (126
mg, 75% yield, 2 steps). .sup.1H NMR (400 MHz, DMSO-d6) .delta.:
9.63 (bs, 1H), 7.53 (d, 1H), 7.26-6.94 (m, 2H), 6.67 (s, 1H),
6.34-6.27 (m, 1H), 6.10 (m, 2H), 5.68 (d, J=10.4, 1H), 5.08 (m,
1H), 4.30 (m, 2H), 3.93 (m, 6H), 3.52 (m, 2H), 3.29 (m, 1H), 3.15
(m, 1H), 3.06 (m, 1H), 2.47 (m, 1H), 1.98 (m, 1H), 0.87 (m, 2H),
0.64 (m, 2H). ESI-MS m/z: 462.5 [M+H].sup.+.
Example 57
Synthesis of
1-(3-(4-(2-(4-chloro-5-cyclopropyl-2-hydroxyphenylamino)acetyl)-2-(hydrox-
ymethyl)piperazin-1-yl)azetidin-1-yl)prop-2-en-1-one (II-54)
##STR00674##
[0918] 1-tert-Butyl 3-methyl
4-(1-benzhydrylazetidin-3-yl)piperazine-1,3-dicarboxylate
[0919] A mixture of 1-benzhydrylazetidin-3-yl methanesulfonate (2.4
g, 7.56 mmol), tert-butyl methyl piperazine-1,3-dicarboxylate (1.85
g, 7.56 mmol), K.sub.2CO.sub.3 (1.6 g, 11.34 mmol) in CH.sub.3CN
(40 mL) was stirred at reflux for 16 h. The mixture was partitioned
between ethyl acetate and water. The organic layer was washed with
water and brine, dried over Na.sub.2SO.sub.4, and concentrated in
vacuo. The residue was purified by flash column chromatography on
silica gel (10% petroleum ether/ethyl acetate) to afford the
desired product (1.85 g, 51% yield).
tert-Butyl
4-(1-benzhydrylazetidin-3-yl)-3-(hydroxymethyl)piperazine-1-car-
boxylate
[0920] To a mixture of LiAlH.sub.4 (500 mg, 13.5 mmol) in THF (40
mL) at -40.degree. C. under argon, a solution of 1-tert-butyl
3-methyl 4-(1-benzhydrylazetidin-3-yl) piperazine-1,3-dicarboxylate
(1.8 g, 3.87 mmol) in THF (10 mL) was added dropwise. The reaction
mixture was stirred at -5.degree. C. to 5.degree. C. for 1 h and
cooled to -20.degree. C. Then water (2 mL) and NaOH (15%) aqueous
were added. The resulting mixture was stirred for 15 min. The solid
was filtered, and the cake rinsed with ethyl acetate. The combined
filtrate was dried over Na.sub.2SO.sub.4 and concentrated in vacuo
to afford the product (1.6 g, 94% yield).
1-(3-(4-(2-(4-Chloro-5-cyclopropyl-2-hydroxyphenylamino)acetyl)-2-(hydroxy-
methyl)piperazin-1-yl)azetidin-1-yl)prop-2-en-1-one (II-54)
[0921] The title compound was prepared from
4-(2-(4,5-dichloro-2-methoxyphenylamino)acetyl)piperazine-2-carbonitrile
in four steps according to the procedure described in Example 47.
.sup.1H NMR (400 MHz, DMSO-d6) .delta.: 9.68 (bs, 1H), 6.67 (s,
1H), 6.35-6.27 (m, 1H), 6.12-6.05 (m, 2H), 5.67 (dd, J=1.6, 10.4
Hz, 1H), 5.11 (m, 1H), 4.82-4.63 (m, 1H), 4.24 (m, 1H), 4.13 (m,
1H), 3.95 (m, 1H), 3.88 (m, 2H), 3.85 (m, 1H), 3.77-3.67 (m, 2H),
3.20 (m, 1H), 3.15 (m, 1H), 2.76-2.60 (m, 2H), 2.40 (m, 1H), 1.95
(m, 1H), 0.87 (m, 2H), 0.62 (m, 2H). ESI-MS m/z: 449.4
[M+H].sup.+.
Example 58
Synthesis of
1-(3-(4-(2-(5,6-dichloro-1h-indol-3-yl)acetyl)piperazin-1-yl)azetidin-1-y-
l)prop-2-en-1-one (II-61)
##STR00675##
[0922] Ethyl 2-(5,6-dichloro-1H-indol-3-yl)acetate
[0923] To a mixture of 5,6-dichloro-1H-indole (1.0 g, 5.37 mmol),
Cu(OTf).sub.2 (194 mg, 0.537 mmol) in DCM (15 mL) at RT, ethyl
2-diazoacetate (918 mg, 8.05 mmol) was added dropwise. The
resulting mixture was stirred at RT for 16 h, quenched with water,
and then extracted dichloromethane. The organic layer was dried
over Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The
residue was purified by Prep-HPLC to afford the desired product
(120 mg, 8.2% yield) as light yellow solid. ESI-MS m/z: 272.1
[M+H].sup.+.
2-(5,6-Dichloro-1H-indol-3-yl)acetic acid
[0924] A mixture of ethyl 2-(5,6-dichloro-1H-indol-3-yl)acetate
(120 mg, 0.44 mmol), LiOH (90 mg, 2.20 mmol) in THF (3 mL) and
H.sub.2O (1 mL) was stirred at RT for 16 h. The solution was poured
into water, adjusted pH to 3-4 with 1N HCl and extracted with ethyl
acetate. The organic layer was dried over Na.sub.2SO.sub.4 and
concentrated in vacuo to afford the desired product (90 mg, 84.5%
yield) as a yellow solid.
1-(3-(4-(2-(5,6-Dichloro-1H-indol-3-yl)acetyl)piperazin-1-yl)azetidin-1-yl-
)prop-2-en-1-one (II-61)
[0925] A mixture of 2-(5,6-dichloro-1H-indol-3-yl)acetic acid (90
mg, 0.372 mmol), 1-(3-(piperazin-1-yl)azetidin-1-yl)prop-2-en-1-one
(87 mg, 0.446 mmol), EDCI.HCl (107 mg, 0.558 mmol), HOBt (75 mg,
0.558 mmol) in DMF (3 mL) at 0.degree. C., Et.sub.3N (112 mg, 1.11
mmol) was added. The resulting mixture was stirred at RT for 16 h.
The mixture was partitioned between ethyl acetate and water. The
organic layer was washed with brine, dried over Na.sub.2SO.sub.4
and concentrated in vacuo. The residue was purified by flash column
chromatography on silica gel (dichloromethane/methanol=30:1) to
afford the desired product (12 mg, 7.66% yield) as an off-white
solid. .sup.1H NMR (400 MHz, DMSO-d6) .delta.:11.19 (bs, 1H), 7.79
(s, 1H), 7.59 (s, 1H), 7.35 (d, 1H), 6.28 (dd, J=9.6, 16.8 Hz, 1H),
6.09 (dd, J=2.4, 17.2 Hz, 1H), 5.66 (dd, J=2.4, 10.4 Hz, 1H),
4.21-4.18 (m, 1H), 4.02-3.98 (m, 1H), 3.93-3.88 (m, 1H), 3.78 (s,
2H), 3.74-3.70 (m, 1H), 3.53-3.47 (m, 4H), 3.10-3.07 (m, 1H),
2.25-2.19 (m, 4H). ESI-MS m/z: 423.3 [M+1].sup.+.
Example 59
Synthesis of
1-(3-(4-(2-(5-chloro-4-ethyl-2-hydroxyphenylamino)acetyl)piperazin-1-yl)a-
zetidin-1-yl)prop-2-en-1-one (II-52)
##STR00676##
[0926] tert-Butyl
3-(4-(2-(4-bromo-5-chloro-2-methoxyphenylamino)acetyl)piperazin-1-yl)azet-
idine-1-carboxylate
[0927] The title compound was prepared from
4-bromo-5-chloro-2-methoxybenzenamine in three steps according to
the procedure described in Example 43.
tert-Butyl
3-(4-(2-(5-chloro-4-ethyl-2-methoxyphenylamino)acetyl)piperazin-
-1-yl)azetidine-1-carboxylate
[0928] To a mixture of tert-butyl
3-(4-(2-(4-bromo-5-chloro-2-methoxyphenylamino)acetyl)piperazin-1-yl)azet-
idine-1-carboxylate (100 mg, 0.193 mmol), Pd(dppf).sub.2Cl.sub.2
(29 mg, 0.04 mmol) and K.sub.2CO.sub.3 (55 mg, 0.386 mmol) in DMF
(10 mL) at RT, Et.sub.2Zn (0.8 mL, 0.8 mmol, 1.0 M in hexane) was
added. The resulting mixture was stirred at 80.degree. C. for 16 h.
The mixture was partitioned between ethyl acetate and water. The
organic layer was washed with brine, dried over Na.sub.2SO.sub.4
and concentrated in vacuo. The residue was purified by flash column
chromatography on silica gel (dichloromethane/methanol=50:1) to
afford the crude product (100 mg). ESI-MS m/z: 467.5
[M+1].sup.+.
1-(3-(4-(2-(5-Chloro-4-ethyl-2-hydroxyphenylamino)acetyl)piperazin-1-yl)az-
etidin-1-yl)prop-2-en-1-one (V-52)
[0929] The title compound was prepared from tert-butyl
3-(4-(2-(5-chloro-4-ethyl-2-methoxyphenylamino)acetyl)piperazin-1-yl)azet-
idine-1-carboxylate in 3 steps according to the procedure described
in Example 33. .sup.1H NMR (400 MHz, DMSO-d6) .delta.: 9.6 (s, 1H),
6.6 (s, 1H), 6.5 (s, 1H), 6.3 (dd, J=10.4, 17.2 Hz, 1H), 6.1 (dd,
J=2.4, 17.2 Hz, 1H), 5.7 (dd, J=2.4, 10.4 Hz, 1H), 5.1 (t, J=4.4
Hz, 1H), 4.2 (t, J=8 Hz, 1H), 4.1 (dd, J=4.8, 8.8 Hz, 1H), 3.95
(dd, J=7.2, 10.0 Hz, 1H), 3.9 (d, J=4.4 Hz, 2H), 3.8 (dd, J=4.8,
10.4 Hz, 1H), 3.6-3.5 (m, 4H), 3.2-3.1 (m, 1H), 3.1-3.0 (m, 1H),
2.5-2.3 (m, 4H), 1.1 (t, J=7.2 Hz, 3H); ESI-MS m/z: 407.4
[M+H].sup.+.
Example 60
Synthesis of
1-(3-(4-(2-(4-chloro-5-ethyl-2-hydroxyphenylamino)acetyl)piperazin-1-yl)a-
zetidin-1-yl)prop-2-en-1-one (II-55)
##STR00677##
[0930]
tert-Butyl3-(4-(2-((4-chloro-5-ethyl-2-methoxyphenyl)amino)acetyl)p-
iperazin-1-yl) azetidine-1-carboxylate
[0931] The title compound was prepared from tert-butyl
3-(4-(2-(4-chloro-5-iodo-2-methoxyphenylamino)acetyl)piperazin-1-yl)azeti-
dine-1-carboxylate in one step according to the procedure described
in Example 52.
1-(3-(4-(2-(4-chloro-5-ethyl-2-hydroxyphenylamino)acetyl)piperazin-1-yl)az-
etidin-1-yl)prop-2-en-1-one (II-55)
[0932] The title compound was prepared from
tert-butyl3-(4-(2-((4-chloro-5-ethyl-2-methoxyphenyl)amino)acetyl)piperaz-
in-1-yl) azetidine-1-carboxylate in three steps according to the
procedure described in Example 33. .sup.1H NMR (400 MHz, DMSO-d6)
.delta.:9.67 (s, 1H), 6.66 (s, 1H), 6.47 (s, 1H), 6.30 (dd, J=10.5,
16.9 Hz, 1H), 6.12 (dd, J=1.7, 16.7 Hz, 1H), 5.69 (dd, J=1.7, 16.7
Hz, 1H), 5.1 (m, 1H), 4.26 (m, 1H), 4.07 (m, 1H), 3.96 (m, 1H),
3.88 (d, J=4.4, 2H), 3.78 (m, 1H), 3.53 (m, 4H), 3.17 (m, 1H), 2.54
(m, 2H), 2.37 (m, 4H), 1.14 (m, 3H). ESI-MS m/z:
407.3[M+H].sup.+.
Example 61
Synthesis of
1-(3-(4-(2-(4-chloro-2-hydroxy-5-(2,2,2-trifluoroethyl)phenylamino)acetyl-
)piperazin-1-yl)azetidin-1-yl)prop-2-en-1-one (II-57)
##STR00678##
[0933] (2-Chloro-4-methoxy-5-nitrophenyl)methanol
[0934] To a solution of 2-chloro-4-methoxy-5-nitrobenzaldehyde (6.0
g, 29 mmol) in MeH at 0.degree. C. (50 mL), sodium borohydride
(4.45 g, 117 mmol) was added in portions and the resulting mixture
was stirred at RT for 30 min. The mixture was concentrated in
vacuo. The residue was dissolved in ethyl acetate, washed with
brine, dried over Na.sub.2SO.sub.4 and concentrated in vacuo. The
residue was purified by flash column chromatography on silica gel
(petroleum ether/ethyl acetate=10:1) to afford the desired product
(5.0 g, 78.40 yield).
1-(Bromomethyl)-2-chloro-4-methoxy-5-nitrobenzene
[0935] To a solution of (2-chloro-4-methoxy-5-nitrophenyl)methanol
(5.0 g, 23 mmol) in dichloromethane (50 mL) at 0.degree. C.,
tribromophosphine (3.08 g, 11.5 mmol) was added in portions and the
resulting mixture was stirred at RT for 2 h. The mixture was poured
into ice-water and extracted with dichloromethane. The organic
layer was washed with brine, dried over Na.sub.2SO.sub.4 and
concentrated in vacuo. The residue was purified by flash column
chromatography on silica gel (petroleum ether/ethyl acetate=10:1)
to afford the desired product (3.5 g, 54.2% yield).
1-Chloro-5-methoxy-4-nitro-2-(2,2,2-trifluoroethyl)benzene
[0936] A mixture of (2-chloro-4-methoxy-5-nitrophenyl)methanol (3.5
g, 12.5 mmol), methyl 2,2-difluoro-2-(fluorosulfonyl)acetate (4.8
g, 25 mmol), copper iodide (617 mg, 3.25 mmol) in NMP (20 mL) was
stirred at 80.degree. C. for 24 h under Argon. After cooled to RT,
the reaction mixture was dissolved in ethyl acetate, washed with
brine, dried over Na.sub.2SO.sub.4 and concentrated in vacuo. The
residue was purified by flash column chromatography on silica gel
(petroleum ether/ethyl acetate=100:1) to afford the desired product
(1.2 g, 36.4% yield). .sup.1H NMR (400 MHz, DMSO-d6) .delta.: 8.15
(s, 1H), 7.60 (s, 1H), 3.98 (s, 3H), 3.89 (dd, J=1.7, 11.2 Hz,
2H).
4-Chloro-2-methoxy-5-(2,2,2-trifluoroethyl)aniline
[0937] A mixture of
1-chloro-5-methoxy-4-nitro-2-(2,2,2-trifluoroethyl)benzene (1.2 g,
4.51 mmol), tin(II) chloride dehydrate (5.0 g, 22.5 mmol) in EtOH
(20 mL) was stirred at reflux for 2 h. After cooled to RT, the
reaction mixture was added saturated NaHCO.sub.3 solution to
adjusted pH to 7-8 and then extracted with ethyl acetate. The
organic layer washed with brine, dried over Na.sub.2SO.sub.4 and
concentrated in vacuo to afford the desired product (900 mg, 85%
yield). .sup.1H NMR (400 MHz, DMSO-d6) .delta.: 7.29 (s, 1H), 7.11
(s, 1H), 5.41 (s, 2H), 4.21 (s, 3H), 3.99 (dd, J=1.7, 11.2 Hz,
2H).
1-(3-(4-(2-((4-Chloro-2-hydroxy-5-(2,2,2-trifluoroethyl)phenyl)amino)acety-
l)piperazin-1-yl)azetidin-1-yl)prop-2-en-1-one (II-57)
[0938] The title compound was prepared from
4-chloro-2-methoxy-5-(2,2,2-trifluoroethyl)aniline in six steps
according to the procedure described in Example 48. .sup.1H NMR
(400 MHz, DMSO-d6) .delta.: 10.08 (s, 1H), 6.74 (s, 1H), 6.59 (s,
1H), 6.34 (dd, J=10.5, 16.9 Hz, 1H), 6.12 (dd, J=1.7, 16.7 Hz, 1H),
5.69 (dd, J=1.7, 16.7 Hz, 1H), 5.22 (m, 1H), 4.24 (m, 1H), 4.04 (m,
1H), 3.94 (m, 1H), 3.88 (d, J=4.4 Hz, 2H), 3.78 (m, 1H), 3.57 (m,
2H), 3.54 (m, 4H), 3.18 (m, 1H), 2.37 (m, 4H). ESI-MS m/z:
461.2[M+H].sup.+.
Example 62
Synthesis of
(e)-1-(4-(1-acryloylazetidin-3-yl)piperazin-1-yl)-3-(4-chloro-5-cycloprop-
yl-2-hydroxyphenyl)prop-2-en-1-one (II-62)
##STR00679##
[0939] 4-Chloro-5-cyclopropyl-2-methoxybenzenamine
[0940] A mixture of 4-chloro-5-iodo-2-methoxyaniline (5.0 g, 17.6
mmol), cyclopropylboronic acid (1.8 g, 21.1 mmol), Pd(OAc).sub.2
(314 mg, 1.4 mmol), tricyclohexylphosphine (500 mg, 17.6 mmol),
K.sub.3PO.sub.4.3H.sub.2O (16.4 g, 61.6 mmol) in toluene (62.5 mL)
and H.sub.2O (3 mL) was stirred at 80.degree. C. under argon for 16
h. The mixture was allowed to cool to RT, and then partitioned
between ethyl acetate and water. The organic layer was washed with
brine, dried over Na.sub.2SO.sub.4 and concentrated in vacuo. The
residue was purified by flash column chromatography on silica gel
(petroleum ether/ethyl acetate=10:1) to afford the desired product
(3.1 g, 88.5% yield). ESI-MS m/z: 198.2[M+H].sup.+.
1-Chloro-2-cyclopropyl-4-iodo-5-methoxybenzene
[0941] To a mixture of 4-chloro-5-cyclopropyl-2-methoxyaniline (2.2
g, 11.05 mmol), conc. HCl (12 mL) and water (12 mL) at 0.degree.
C., the solution of sodium nitrate (762.8 mg, 11.05 mmol) in water
(2.5 mL) was added dropwise. After stirring at 0.degree. C. for 15
min, a solution of KI (1.83 g, 11.05 mmol) in water (5 mL) was
added dropwise. The resulting mixture was stirred at RT for 4 h,
poured into water (20 mL) and then extracted with dichloromethane.
The organic layer was washed with brine, dried over anhydrous
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The residue
was purified by flash column chromatography on silica gel (0-10%
ethyl acetate/petroleum ether) to afford the desired product (680
mg, 20% yield) as a solid. .sup.1H NMR (400 MHz, DMSO-d6) .delta.:
7.37 (s, 1H), 7.08 (s, 1H), 3.84 (s, 3H), 2.00 (m, 1H), 0.89 (m,
2H), 0.65 (m, 1H).
(E)-1-(4-(1-Acryloylazetidin-3-yl)piperazin-1-yl)-3-(4-chloro-5-cyclopropy-
l-2-methoxyphenyl)prop-2-en-1-one
[0942] A mixture of 1-chloro-2-cyclopropyl-4-iodo-5-methoxybenzene
(300 mg, 0.974 mmol), tert-butyl
3-(4-acryloylpiperazin-1-yl)azetidine-1-carboxylate (431 mg, 1.46
mmol), Pd(OAc).sub.2 (54.6 mg, 0.243 mmol), sodium acetate (239 mg,
2.92 mmol), tetrabutylammonium chloride (539 mg, 1.95 mmol) in DMF
(7 mL) was stirred at 100.degree. C. for 24 h. The mixture was
partitioned between ethyl acetate and water. The organic layer was
washed with brine, dried over Na.sub.2SO.sub.4, filtered and
concentrated in vacuo to purified by silica gel
(dichloromethane/methanol=40:1) to afford the desired product (350
mg, 84% yield). ESI-MS m/z: 476.2 [M+H].sup.+.
(E)-1-(4-(1-Acryloylazetidin-3-yl)piperazin-1-yl)-3-(4-chloro-5-cyclopropy-
l-2-hydroxyphenyl)prop-2-en-1-one (II-62)
[0943] The title compound was prepared from
(E)-1-(4-(1-acryloylazetidin-3-yl)piperazin-1-yl)-3-(4-chloro-5-cycloprop-
yl-2-methoxyphenyl)prop-2-en-1-one in three steps according to the
procedure described in Example 33. .sup.1H NMR (400 MHz, DMSO-d6)
.delta.: 10.3 (s, 1H), 7.71 (m, 1H), 7.30 (s, 1H), 7.22 (m, 1H),
6.93 (s, 1H), 6.34 (dd, J=10.5, 16.9 Hz, 1H), 6.12 (dd, J=1.7, 16.7
Hz, 1H), 5.66 (dd, J=1.7, 16.7 Hz, 1H), 4.26 (m, 1H), 4.08 (m, 1H),
3.94 (m, 1H), 3.79 (m, 1H), 3.69 (m, 2H), 3.58 (m, 2H), 3.18 (m,
1H), 2.33 (m, 4H), 1.99 (m, 1H), 0.92 (m, 2H), 0.73 (m, 2H). ESI-MS
m/z: 416 [M+H].sup.+.
Example 63
Synthesis of
1-(3-(4-(2-(4-chloro-5-cyclopropyl-2-methoxyphenylthio)acetyl)piperazin-1-
-yl)azetidin-1-yl)prop-2-en-1-one (II-65)
##STR00680##
[0944] Methyl
2-((4-chloro-5-cyclopropyl-2-methoxyphenyl)thio)acetate
[0945] A mixture of 1-chloro-2-cyclopropyl-4-iodo-5-methoxybenzene
(380 mg, 1.23 mmol), Pd.sub.2(dba).sub.3 (56 mg, 0.061 mmol),
methyl 2-mercaptoacetate (196 mg, 1.85 mmol),
1,1'-Bis(diphenylphosphino)ferrocene (136 mg, 0.246 mmol),
Et.sub.3N (372 mg, 3.69 mmol) in NMP (8 mL) was stirred under argon
at 80.degree. C. for 24 h. After cooled to RT, the reaction mixture
was extracted with ethyl acetate. The organic layer was washed with
brine, dried over Na.sub.2SO.sub.4 and concentrated in vacuo. The
residue was purified by flash column chromatography on silica gel
(petroleum ether/ethyl acetate=20:1) to afford the desired product
(340 mg, 92% yield). .sup.1H NMR (400 MHz, DMSO-d6) .delta.: 7.05
(s, 1H), 6.81 (s, 1H), 3.84 (s, 2H), 3.82 (s, 3H), 3.62 (s, 3H),
2.00 (m, 1H), 0.94 (m, 2H), 0.64 (m, 2H).
1-(3-(4-(2-(4-Chloro-5-cyclopropyl-2-methoxyphenylthio)acetyl)piperazin-1--
yl)azetidin-1-yl)prop-2-en-1-one (II-65)
[0946] The title compound was prepared from methyl
2-((4-chloro-5-cyclopropyl-2-methoxyphenyl)thio)acetate in four
steps according to the procedure described in Example 43. .sup.1H
NMR (400 MHz, DMSO-d6) .delta.: 7.03 (s, 1H), 6.93 (s, 1H), 6.31
(dd, J=10.5, 16.9 Hz, 1H), 6.11 (dd, J=1.7, 16.7 Hz, 1H), 5.68 (dd,
J=1.7, 16.7 Hz, 1H), 4.25 (m, 1H), 4.04 (m, 1H), 3.88 (s, 2H) 3.81
(s, 3H), 3.75 (m, 1H), 3.52 (m, 4H), 3.16 (m, 1H), 2.36-2.25 (m,
4H), 2.02 (m, 1H), 0.93 (m, 2H), 0.66 (m, 2H). ESI-MS m/z: 450
[M+H].sup.+.
Example 64
Synthesis of
(S)-1-(3-(4-(2-(4-chloro-5-cyclopropyl-2-hydroxyphenylamino)propanoyl)pip-
erazin-1-yl)azetidin-1-yl)prop-2-en-1-one (II-67)
##STR00681##
[0947] (S)-2-(5-Bromo-4-chloro-2-methoxyphenylamino)propanoic
acid
[0948] A mixture of 1-bromo-2-chloro-5-iodo-4-methoxybenzene (3 g,
8.64 mmol), (S)-2-aminopropanoic acid (769 mg, 8.64 mmol), CuI (164
mg, 0.864 mmol), 2-hydroxybenzaldehyde phenylhydrazone (366 mg,
1.73 mmol), K.sub.3PO.sub.4.3H.sub.2O (4.6 g, 17.28 mmol) in DMF
(10 mL) was stirred under argon at 80.degree. C. for 16 h. The
mixture was allowed to cool to RT, H.sub.2O and Et.sub.2O were
added to the solution. The resulting solution was partitioned into
two phases, the aqueous phase was separated, and the organic layer
was extracted with 5% NaOH. The combined aqueous phase was
acidified to pH 4 with 20% HCl, and then extracted with Et.sub.2O.
The resulting organic layer was dried over MgSO.sub.4 and
concentrated in vacuo to afford the desired product (1.7 g, 64%
yield). ESI-MS m/z: 306.1 [M+H].sup.-
(S)-tert-Butyl
3-(4-(2-((5-bromo-4-chloro-2-methoxyphenyl)amino)propanoyl)piperazin-1-yl-
)azetidine-1-carboxylate
[0949] To a solution of
(S)-2-(5-bromo-4-chloro-2-methoxyphenylamino)propanoic acid (1.6 g,
5.21 mmol), tert-butyl 3-(piperazin-1-yl)azetidine-1-carboxylate
(1.88 g, 7.82 mmol), EDCI.HCl (2.0 g, 10.42 mmol), HOBt (1.41 g,
10.42 mmol) in DMF (20 mL) at 0.degree. C., Et.sub.3N (1.58 g,
15.63 mmol) was added. The resulting mixture was stirred at RT for
16 h and then partitioned between ethyl acetate and water. The
organic layer was washed with brine, dried over Na.sub.2SO.sub.4
and concentrated in vacuo. The residue was purified by flash column
chromatography on silica gel (methanol/dichloroethane=1:50) to
afford the desired product (2.1 g, 76% yield). ESI-MS m/z: 531.3
[M+H].sup.+.
((S)-tert-Butyl
3-(4-(2-((4-chloro-5-cyclopropyl-2-methoxyphenyl)amino)propanoyl)piperazi-
n-1-yl)azetidine-1-carboxylate
[0950] A mixture of (S)-tert-butyl
3-(4-(2-((5-bromo-4-chloro-2-methoxyphenyl)amino)propanoyl)piperazin-1-yl-
)azetidine-1-carboxylate (700 mg, 1.32 mmol), cyclopropylboronic
acid (114 mg, 1.32 mmol), Pd(OAc).sub.2 (15 mg, 0.066 mmol),
tricyclohexylphosphine (37 mg, 0.132 mmol),
K.sub.3PO.sub.4.3H.sub.2O (974 mg, 4.62 mmol) in DMF (10 mL) and
H.sub.2O (0.5 mL) was stirred under argon at 80.degree. C. for 16
h. The mixture was allowed to cool to RT, and then partitioned
between ethyl acetate and water. The organic layer was washed with
brine, dried over Na.sub.2SO.sub.4 and concentrated in vacuo. The
residue was purified by flash column chromatography on silica gel
(methanol/dichloroethane=1:100) to afford the desired product (400
mg, 62%). ESI-MS m/z: 493.2[M+H].sup.+.
(S)-1-(3-(4-(2-((4-Chloro-5-cyclopropyl-2-hydroxyphenyl)amino)propanoyl)pi-
perazin-1-yl)azetidin-1-yl)prop-2-en-1-one (II-67)
[0951] The title compound was prepared from (S)-tert-butyl
3-(4-(2-((4-chloro-5-cyclopropyl-2-methoxyphenyl)amino)propanoyl)piperazi-
n-1-yl)azetidine-1-carboxylate in three steps according to the
procedure described in Example 33. .sup.1H NMR (400 MHz, DMSO-d6)
.delta.: 9.69 (s, 1H), 6.66 (s, 1H), 6.29 (dd, J=10.5, 16.9 Hz,
1H), 6.12 (dd, J=1.7, 16.7 Hz, 1H), 6.05 (s, 1H), 5.68 (dd, J=1.7,
16.7 Hz, 1H), 4.84 (m, 1H), 4.61 (m, 1H), 4.24 (m, 1H), 4.06 (m,
1H), 3.94 (m, 1H), 3.78 (m, 4H), 3.55 (m, 1H), 2.43-2.17 (m, 4H),
1.97 (m, 1H), 0.88 (m, 2H), 0.63 (m, 2H). ESI-MS m/z: 433.3
[M+H].sup.+.
Example 65
Synthesis of
(s)-1-(3-(4-(2-(4-chloro-5-ethyl-2-hydroxyphenylamino)propanoyl)piperazin-
-1-yl)azetidin-1-yl)prop-2-en-1-one (II-69)
##STR00682##
[0952] (S)-tert-Butyl
3-(4-(2-((4-chloro-5-ethyl-2-methoxyphenyl)amino)propanoyl)piperazin-1-yl-
)azetidine-1-carboxylate
[0953] To a solution of (S)-tert-butyl
3-(4-(2-((5-bromo-4-chloro-2-methoxyphenyl)amino)propanoyl)piperazin-1-yl-
)azetidine-1-carboxylate (400 mg, 0.75 mmol), PdCl.sub.2(dppf) (95
mg, 0.13 mmol) in THF (20 mL) at RT, Et.sub.2Zn (2.86 mL, 2.86
mmol, 1.0 M in hexane) was added. The resulting mixture was stirred
under argon at 80.degree. C. for 4 h and then partitioned between
ethyl acetate and water. The organic layer was washed with brine,
dried over Na.sub.2SO.sub.4 and concentrated in vacuo. The residue
was purified by flash column chromatography on silica gel
(methanol/dichloroethane=1:80) to afford the desired product (250
mg, 69% yield). ESI-MS m/z: 481.2 [M+H].sup.+.
(S)-1-(3-(4-(2-((4-Chloro-5-ethyl-2-hydroxyphenyl)amino)propanoyl)piperazi-
n-1-yl)azetidin-1-yl)prop-2-en-1-one (II-69)
[0954] The title compound was prepared from (S)-tert-butyl
3-(4-(2-((4-chloro-5-ethyl-2-methoxyphenyl)amino)propanoyl)piperazin-1-yl-
)azetidine-1-carboxylate in three steps according to the procedure
described in Example 33. .sup.1H NMR (400 MHz, DMSO-d6) .delta.:
9.61 (s, 1H), 6.64 (s, 1H), 6.48 (s, 1H), 6.29 (dd, J=10.5, 16.9
Hz, 1H), 6.12 (dd, J=1.7, 16.7 Hz, 1H), 5.68 (dd, J=1.7, 16.7 Hz,
1H), 4.89 (m, 1H), 4.61 (m, 1H), 4.25 (m, 1H), 4.06 (m, 1H), 3.94
(m, 1H), 3.72-3.53 (m, 4H), 3.16 (m, 1H), 2.5 (m, 2H), 2.43-2.17
(m, 4H), 1.21 (dd, 3H), 1.15 (m, 3H). ESI-MS m/z: 406.2
[M+H].sup.+.
Example 66
Synthesis of
1-(3-(4-(2-(4-chloro-5-cyclopropyl-2-hydroxyphenylamino)acetyl)-2-methylp-
iperazin-1-yl)azetidin-1-yl)prop-2-en-1-one (II-60)
##STR00683##
[0955] tert-Butyl
4-(1-acryloylazetidin-3-yl)-3-methylpiperazine-1-carboxylate
[0956] The title compound was prepared from tert-butyl
3-methylpiperazine-1-carboxylate in three steps according to the
procedure described in Example 41.
1-(3-(2-Methylpiperazin-1-yl)azetidin-1-yl)prop-2-en-1-one
hydrochloride
[0957] The mixture of tert-butyl
4-(1-acryloylazetidin-3-yl)-3-methylpiperazine-1-carboxylate (62
mg, 0.199 mmol) in MeOH/HCl (20 mL, 2.9 M) was stirred at RT for 1
h. The mixture was concentrated in vacuo to afford the crude
product (59 mg). The crude product was used directly in the next
step without further purification.
1-(3-(4-(2-((4-Chloro-5-cyclopropyl-2-hydroxyphenyl)amino)acetyl)-2-methyl-
piperazin-1-yl)azetidin-1-yl)prop-2-en-1-one (II-60)
[0958] To the mixture of
2-((4-chloro-5-cyclopropyl-2-hydroxyphenyl)amino)acetic acid (30
mg, 0.124 mmol) and NMM (50 mg, 0.496 mmol) in dry THF (30 mL) at
-10.degree. C., ethyl chloroformate (15 mg, 0.136 mmol) was added
and the resulting mixture was stirred at -10.degree. C. for 45 min.
Then it was added a mixture of
1-(3-(2-methylpiperazin-1-yl)azetidin-1-yl)prop-2-en-1-one
hydrochloride (37 mg, 0.149 mmol), Et.sub.3N (50 mg, 0.496 mmol)
and dichloromethane (3 mL). The resulting mixture was stirred at RT
for 30 min. The mixture was partitioned between ethyl acetate and
water. The organic layer was washed with saturated NaHCO.sub.3
solution and brine, dried over Na.sub.2SO.sub.4, filtered and
concentrated in vacuo. The residue was purified with column
chromatography on silica gel (dichloromethane/methanol=40:1) to
afford the desired product (10 mg, 18.6% yield) as a white solid.
.sup.1H NMR (400 MHz, DMSO-d6) .delta.: 9.65 (s, 1H), 6.66 (s, 1H),
6.34-6.27 (m, 1H), 6.10-6.07 (m, 2H), 5.68-5.65 (d, J=10.4 Hz, 1H),
5.12 (m, 1H), 4.29-4.19 (m, 1H), 4.12-4.10 (m, 1H), 4.08-3.81 (m,
4H), 3.78 (s, 4H), 2.63 (m, 2H), 2.25 (m, 1H), 1.96 (m, 1H), 1.24
(s, 1H), 0.96-0.87 (m, 5H), 0.63 (m, 2H). ESI-MS m/z: 433.5
[M+H].sup.-.
Example 67
Synthesis of
2-(2-(4-(1-acryloylazetidin-3-yl)piperazin-1-yl)-2-oxoethylamino)-5-chlor-
o-4-cyclopropylbenzonitrile (II-71)
##STR00684##
[0959] 4-Bromo-5-chloro-2-nitrobenzamide
[0960] A mixture of 4-bromo-5-chloro-2-nitrobenzoic acid (1.3 g,
4.63 mmol), Et.sub.3N (1.4 g, 13.9 mmol) in THF (20 mL) at
0.degree. C., ethyl chloroformate (1.5 g, 13.9 mmol) was added. The
resulting mixture was stirred at 0.degree. C. for 1 h. Then
NH.sub.3.H.sub.2O (4 mL) was added and stirred for 0.5 h. The
mixture was extracted with ethyl acetate. The organic layer was
washed with brine, dried over anhydrous Na.sub.2SO.sub.4, filtered
and concentrated in vacuo to afford the crude product (900 mg).
2-Amino-4-bromo-5-chlorobenzamide
[0961] To a solution of 4-bromo-5-chloro-2-nitrobenzamide (900 mg,
3.2 mmol) in AcOH (20 mL) and water (5 mL) at 70.degree. C., Fe
powder (900 mg, 16.1 mmol) was added and the resulting mixture was
stirred at 70.degree. C. for 1 h. The mixture was allowed to cool
to RT and poured into ice-water. The precipitate was collected by
filtration and rinsed with water. This crude product was dissolved
with ethyl acetate and filtered. The filtrate was washed with
saturated NaHCO.sub.3 solution and brine. The organic layer was
dried over MgSO.sub.4, filtered, and concentrated in vacuo to
afford the desired product (770 mg, 97% yield). ESI-MS m/z: 250.1
[M+H].sup.+.
tert-Butyl
3-(4-(2-((5-bromo-2-carbamoyl-4-chlorophenyl)amino)acetyl)piper-
azin-1-yl)azetidine-1-carboxylate
[0962] The title compound was prepared from
2-amino-4-bromo-5-chlorobenzamide in three steps according to the
procedure described in Example 44. ESI-MS m/z: 532.5
[M+H].sup.+.
tert-Butyl
3-(4-(2-((2-carbamoyl-4-chloro-5-cyclopropylphenyl)amino)acetyl-
)piperazin-1-yl)azetidine-1-carboxylate
[0963] To a solution of tert-butyl
3-(4-(2-((5-bromo-2-carbamoyl-4-chlorophenyl)amino)acetyl)piperazin-1-yl)-
azetidine-1-carboxylate (350 mg, 0.66 mmol) and cyclopropylboronic
acid (226 mg, 2.64 mmol) in toluene (10 mL) and water (2 mL),
Pd(OAc).sub.2(15 mg, 0.07 mmol), PCy.sub.3 (37 mg, 0.132 mmol) and
K.sub.3PO.sub.4 (487 mg, 2.31 mmol) were added. The mixture was
stirred at 80.degree. C. for 16 h. The mixture was allowed to cool
to RT and concentrated in vacuo. The residue was purified by flash
column chromatography on silica gel (1-5% methanol/dichloroethane)
to afford the desired product (150 mg, 46% yield) as a solid.
2-((2-(4-(1-Acryloylazetidin-3-yl)piperazin-1-yl)-2-oxoethyl)amino)-5-chlo-
ro-4-cyclopropylbenzamide
[0964] The title compound was prepared from tert-butyl
3-(4-(2-((2-carbamoyl-4-chloro-5-cyclopropylphenyl)amino)acetyl)piperazin-
-1-yl)azetidine-1-carboxylate in two steps according to the
procedure described in Example 33. ESI-MS m/z: 446.4
[M+H].sup.+.
2-((2-(4-(1-Acryloylazetidin-3-yl)piperazin-1-yl)-2-oxoethyl)amino)-5-chlo-
ro-4-cyclopropylbenzonitrile (II-71)
[0965] A mixture of
2-((2-(4-(1-acryloylazetidin-3-yl)piperazin-1-yl)-2-oxoethyl)amino)-5-chl-
oro-4-cyclopropylbenzamide (3 0 mg, 0.067 mmol) and Et.sub.3N (41
mg, 0.404 mmol) in DCM (10 mL) at RT, trifluoroacetic anhydride (56
mg, 0.268 mmol) was added. The resulting mixture was stirred at RT
for 0.5 h, poured into water and then extracted with
dichloromethane. The organic layer was washed with brine, dried
over Na.sub.2SO.sub.4 and concentrated. The residue was purified by
flash column chromatography on silica gel (1-4%
methanol/dichloroethane) to afford the desired product (20 mg, 72%
yield). .sup.1H NMR (400 MHz, DMSO-d6) .delta.: 7.60 (s, 1H),
6.34-6.30 (m, 1H), 6.27 (s, 1H), 6.12-6.07 (m, 1H), 6.01-5.99 (t,
J=4 Hz, 1H), 5.69-5.65 (m, 1H), 4.26-4.22 (m, 1H), 4.07-4.04 (m,
3H), 3.96-3.92 (m, 1H), 3.80-3.76 (m, 1H), 3.53-3.51 (m, 4H),
3.19-3.13 (m, 1H), 2.45-2.30 (m, 4H), 2.16-2.09 (m, 1H), 1.08-1.03
(m, 2H), 0.87-0.80 (m, 2H). ESI-MS m/z: 428.4 [M+H].sup.+.
Example 68
Synthesis of
1-(3-(4-(2-(4-chloro-5-(2,2-difluorocyclopropyl)-2-hydroxyphenylamino)ace-
tyl)piperazin-1-yl)azetidin-1-yl)prop-2-en-1-one (II-56)
##STR00685## ##STR00686##
[0966] 2-Chloro-4-methoxy-1-vinylbenzene
[0967] To a suspension of phosphonium salt (2.05 g, 5 mmol) in THF
(50 mL), was added t-BuOK (0.84 g, 7.5 mmol). The mixture turned to
yellow and was kept stirring at RT for 1h.
2-Chloro-4-methoxybenzaldehyde (0.85 g, 5 mmol) was added to the
mixture. The mixture was stirred for 24h, diluted with sat.
NaHCO.sub.3 and then extracted with hexane. Organic layer was dried
over Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The
residue was purified by Isolera One (100% hexanes to afford the
desired product (0.45 g, 53% yield). .sup.1H NMR (CDCl.sub.3,
.delta.): 7.49 (d, J=6.8 Hz, 1H), 7.03 (dd, J=8.8, 14.0 Hz, 1H),
6.90 (d, J=2.0, 1H), 6.79 (dd, J=2.0, 6.8 Hz, 1H), 5.62 (d, J=14.0
Hz, 1H), 5.26 (d, J=8.8 Hz, 1H), 3.80 (s, 3H).
2-Chloro-1-(2,2-difluorocyclopropyl)-4-methoxybenzene
[0968] The solution of 2-chloro-4-methoxy-1-vinylbenzene (290 mg,
1.72 mmol) in dry THF (4 mL) was degassed, and then TMS-CF.sub.3
and NaI were added. The mixture was stirred at 80.degree. C.
overnight. TLC (100% Hexane) showed the reaction as complete. The
mixture was diluted with hexane (20 mL). The inorganic salt was
removed by filtration. The filtrate was concentrate in vacuo. The
residue was purified via Isolera One (Hexane=100%).
1-Chloro-2-(2,2-difluorocyclopropyl)-5-methoxy-4-nitrobenzene
[0969] To a solution of
2-chloro-1-(2,2-difluorocyclopropyl)-4-methoxybenzene (328 mg, 1.5
mmol) in Ac.sub.2O (2 mL), was added HNO.sub.3 (10 drops) at
0.degree. C. The mixture was stirred from 0.degree. C. to rt.
Ac.sub.2O was removed in vacuo. The residue was diluted with DCM
and washed with water. The organic layer was dried over
Na.sub.2SO.sub.4. The solvent was removed in vacuo. The residue was
purified via Isolera One (EtOAc/Hexane=0-15%) to afford the desired
product. .sup.1H NMR (CDCl.sub.3, .delta.): 7.77 (s, 1H), 7.16 (s,
1H), 3.98 (s, 3H), 2.78-2.90 (m, 1H), 1.90-1.98 (m, 1H), 1.60-1.68
(m, 1H). ESI-MS m/z: 264.1 [M+H].sup.+.
4-Chloro-5-(2,2-difluorocyclopropyl)-2-methoxyaniline
[0970] The above obtained
1-chloro-2-(2,2-difluorocyclopropyl)-5-methoxy-4-nitrobenzene was
dissolved in 10 mL of co-solvent of AcOH/i-PrOH (1:5). Zn dust was
added to the mixture. The mixture was stirred at 60.degree. C. for
30 min. The solvent was removed in vacuo. The residue was diluted
was DCM and the inorganic salt was removed by filtration. The
filtrate was concentrated to give crude product which was used in
next step without further purification.
1-(3-(4-((4-chloro-5-(2,2-difluorocyclopropyl)-2-hydroxyphenyl)glycyl)pipe-
razin-1-yl)azetidin-1-yl)prop-2-en-1-one (II-56)
[0971] The title compound was prepared from
4-chloro-5-(2,2-difluorocyclopropyl)-2-methoxyaniline in 6 steps
according to the procedure described in Example 44. .sup.1H NMR
(CDCl.sub.3, .delta.): 9.90 (s, 1H), 6.73 (s, 1H), 6.40 (s, 1H),
6.30 (dd, J=8.4, 13.6 Hz, 1H), 6.10 (dd, J=1.6, 12.0 Hz, 1H), 5.66
(dd, J=1.6, 8.4 Hz, 1H), 5.18 (t, J=3.2, 3.6 Hz, 1H), 4.24 (t,
J=6.0, 6.8 Hz, 1H), 4.03-4.08 (m, 1H), 3.86-3.97 (m, 3H), 3.74-3.80
(m, 1H), 3.52 (br. s, 4H), 3.13-3.20 (m, 1H), 2.77-2.87 (m, 1H),
2.25-2.43 (m, 4H), 1.87-1.97 (m, 2H). ESI-MS m/z: 455.2
[M+H].sup.+.
Example 69
Biochemical Assay of Compounds of Structure (I), (II) and (III)
[0972] Test compounds were prepared as 10 mM stock solutions in
DMSO (Fisher cat# BP-231-100). KRAS G12C 1-169, his-tagged protein,
GDP-loaded was diluted to 2 .mu.m in buffer (20 mM Hepes, 150 mM
NaCl, 1 mM MgCl.sub.2). Compounds were tested for activity as
follows:
[0973] Compounds were diluted to 50.times. final test concentration
in DMSO in 96-well storage plates. Compound stock solutions were
vortexed before use and observed carefully for any sign of
precipitation. Dilutions were as follow: [0974] For 100 .mu.M final
compound concentration, compounds were diluted to 5000 .mu.M (5
.mu.l 10 mM compound stock+5 .mu.l DMSO and mixed well by
pipetting. [0975] For 30 .mu.M final compound concentration,
compounds were diluted to 1500 .mu.M (3 .mu.l 10 mM compound
stock+17 .mu.l DMSO) and mixed well by pipetting. [0976] For 10
.mu.M final compound concentration, compounds were diluted to 500
.mu.M (2 .mu.l 10 mM compound stock+38 .mu.l DMSO) and mixed well
by pipetting. 49 .mu.l of the stock protein solution was added to
each well of a 96-well PCR plate (Fisher cat#1423027). 1 .mu.l of
the diluted 50.times. compounds were added to appropriate wells in
the PCR plate using 12-channel pipettor. Reactions were mixed
carefully and thoroughly by pipetting up/down with a 200 .mu.l
multi-channel pipettor. The plate was sealed well with aluminum
plate seal, and stored in drawer at room temperature for 24 hrs. 5
.mu.l of 2% formic acid (Fisher cat# A117) in DI H.sub.2O was then
added to each well followed by mixing with a pipette. The plate was
then resealed with aluminum seal and stored on dry ice until
analyzed as described below.
[0977] The above described assays were analyzed by mass
spectrometry according to the following procedure:
[0978] The MS instrument is set to positive polarity, 2 GHz
resolution, and low mass (1700) mode and allowed to equilibrate for
30 minutes. The instrument is then calibrated, switched to
acquisition mode and the appropriate method loaded.
[0979] After another 30 minute equilibration time, a blank batch
(i.e., buffer) is run to ensure equipment is operating properly.
The samples are thawed at 37.degree. C. for 10 minutes, briefly
centrifuged, and transfer to the bench top. Wells A1 and H12 are
spiked with 1 .mu.L 500 M internal standard peptide, and the plates
centrifuged at 2000.times.g for 5 minutes. The method is then run
and masses of each individual well recorded.
[0980] The masses (for which integration data is desired) for each
well are pasted into the platemap and exported from the analysis.
Masses for the internal standards are exported as well. The data at
50 ppm is extracted for the +19 charge state, and identity of well
A1 is assigned using the internal standard spike and integrated.
Peak data is exported as a TOF list and the above steps are
repeated individually, for the +20, 21, 22, 23, 24, and charge
states.
[0981] Other in vitro analyses are as follows:
Inhibition of Cell Growth:
[0982] The ability of the subject compounds to inhibit Ras-mediated
cell growth is assessed and demonstrated as follows. Cells
expressing a wildtype or a mutant Ras are plated in white, clear
bottom 96 well plates at a density of 5,000 cells per well. Cells
are allowed to attach for about 2 hours after plating before a
compound disclosed herein is added. After certain hours (e.g., 24
hours, 48 hours, or 72 hours of cell growth), cell proliferation is
determined by measuring total ATP content using the Cell Titer Glo
reagent (Promega) according to manufacturer's instructions.
Proliferation EC50s is determined by analyzing 8 point compound
dose responses at half-log intervals decreasing from 100 M.
Inhibition of Ras-Mediated Signaling Transduction:
[0983] The ability of the compounds disclosed herein in inhibiting
Ras-mediated signaling is assessed and demonstrated as follows.
Cells expressing wild type or a mutant Ras (such as G12C, G12V, or
G12A) are treated with or without (control cells) a subject
compound. Inhibition of Ras signaling by one or more subject
compounds is demonstrated by a decrease in the steady-state level
of phosphorylated MEK, and/or Raf binding in cells treated with the
one or more of the subject compounds as compared to the control
cells.
Inhibition of Ras-Mediated Signaling Transduction:
[0984] The ability of the compounds disclosed herein in inhibiting
Ras-mediated signaling is assessed and demonstrated as follows.
Cells expressing wild type or a mutant Ras (such as G12C, G12V, or
G12A) are treated with or without (control cells) a subject
compound. Inhibition of Ras signaling by one or more subject
compounds is demonstrated by percentage binding of compound to the
G12C mutated Ras protein in cells treated with the one or more of
the subject compounds as compared to the control cells.
Inhibition of Ras-Mediated Signaling Transduction:
[0985] The ability of the compounds disclosed herein in inhibiting
Ras-mediated signaling is assessed and demonstrated as follows.
Cells expressing wild type or a mutant Ras (such as G12C, G12V, or
G12A) are treated with or without (control cells) a subject
compound. Inhibition of Ras signaling by one or more subject
compounds is demonstrated by a decrease in binding of Ras complex
to downstream signaling molecules (for example Raf) in cells
treated with the one or more of the subject compounds as compared
to the control cells.
[0986] Each of the compounds in Tables 1, 2a and 3 were tested
according to the above methods and found to covalently bind to KRAS
G12C to the extent of at least about 5% (i.e., at least about 5% of
the protein present in the well was found to be covalently bound to
test compound).
TABLE-US-00005 TABLE 4 Activity of Representative Compounds of
Structure (I)* Binding No. % No. Binding % No. Binding % No.
Binding % 1 +++ 2 + 3 + 4 ++++ 5 +++ 6 +++ 7 ++++ 8 ++ 9 +++ 10 ++
11 ++++ 12 + 13 ++ 14 ++ 15 +++ 16 ++ 17 + 18 ++ 19 ++ 20 + 21 +++
22 + 23 ++ 24 + 25 ++ 26 ++ 27 ++ 28 + 29 + 30 + 31 + 32 + 33 ++ 34
+++ 35 + 36 + 37 ++ 38 ++ 39 + 40 +++ 41 + 42 +++ 43 +++ 44 +++ 45
+ 46 ++++ 47 ++++ 48 ++++ 49 ++++ 50 + 51 ++++ 52 ++++ 53 ++++ 54
++ 55 ++++ 56 ++++ 57 + 58 +++ 59 ++++ 60 + 61 + 62 + 63 + 64 ++ 65
+ 66 ++++ 67 +++ 68 + 69 + 70 +++ 71 + 72 ++ 73 ++ 74 +++ 75 + 76 +
77 +++ 78 + 79 + 80 + 81 +++ 82 + 83 ++ 84 ++ 85 +++ 86 + 87 + 88 +
89 + 90 + 91 ++ 92 + 93 + 94 ++ 95 ++ 96 + 97 ++ 98 + 99 +++ 100
+++ 101 +++ 102 +++ 103 ++ 104 +++ 105 + 106 ++++ 107 ++++ 108 ++
109 ++++ 110 +++ 111 +++ 112 +++ 113 +++ 114 +++ 115 +++ 116 ++ 117
+++ 118 +++ 119 +++ 120 +++ 121 ++++ 122 ++ 123 ++++ 124 +++ 125
++++ 126 ++++ 127 ++++ 128 +++ 129 + 130 + 131 + 132 +++ 133 ++ 134
++ 135 + 136 ++ 137 + 138 ++++ 139 ++++ 140 +++ 141 + 142 + 143 +++
144 + 145 ++++ 146 ++++ 147 ++ 148 +++ 149 + 150 ++++ 151 + 152 +++
153 ++++ 154 ++ 155 ++ 156 ++ 157 + 158 ++++ 159 +++ 160 + 161 ++
162 + 163 + 164 + 165 N/A 166 + 167 + 168 ++ 169 +++ 170 + 171 +++
172 +++ 173 ++++ 174 ++ 175 +++ 176 +++ 177 + 178 + 179 + 180 + 181
+ 182 +++ 183 +++ 184 ++++ 185 ++ 186 +++ 187 +++ 188 +++ 189 ++
190 + 191 +++ 192 ++ 193 ++ 194 ++++ 195 ++++ 196 ++++ 197 ++ 198
++++ 199 N/A 200 ++ 201 +++ 202 +++ 203 ++++ 204 +++ 205 + 206 +++
207 ++++ 208 ++++ 209 ++++ 210 ++++ 211 + 212 ++++ 213 ++ 214 + 215
++ 216 + 217 +++ 218 +++ 219 + 220 +++ 221 ++ 222 +++ 223 + 224 +++
225 ++ 226 + 227 ++++ 228 + 229 ++ 230 + 231 +++ 232 + 233 +++ 234
++++ 235 +++ 236 +++ 237 ++++ 238 +++ 239 +++ 240 +++ 241 + 242
++++ 243 ++++ 244 + 245 + 246 ++++ 247 +++ 248 N/A 249 + 250 ++ 251
++++ 252 ++++ 253 ++++ 254 +++ 255 +++ 256 +++ 257 ++++ 258 ++ 259
+++ 260 ++ 261 + 262 + 263 + 264 ++ 265 + 266 +++ 267 + 268 +++ 269
+++ 270 +++ 271 +++ 272 ++++ 273 ++++ 274 ++++ 275 ++ 276 + 277 +
278 ++ 279 +++ 280 +++ 281 ++ 282 +++ 283 ++ 284 ++++ 285 +++ 286 +
287 ++ 288 ++ 289 +++ 290 +++ 291 ++++ 292 + 293 ++++ 294 ++++ 295
+ 296 + 297 + 298 ++ 299 + 300 ++ 301 ++ 302 +++ 303 ++ 304 ++ 305
++ 306 ++ 307 ++ 308 +++ 309 +++ 310 ++++ 311 +++ 312 ++++ 313 +++
314 ++++ 315 + 316 ++ 317 N/A 318 + 319 ++ 320 ++ 321 + 322 +++ 323
+++ 324 + 325 ++ 326 + 327 ++ 328 + 329 ++ 330 ++ 331 ++ 332 ++ 333
+ 334 ++++ 335 ++++ 336 +++ 337 + 338 ++ 339 ++++ 340 ++++ 341 ++++
342 ++++ N/A N/A N/A N/A *Binding for compounds 1-47 was measured
at 24 h; binding for compounds 48-246 was measured at 2 h; binding
for compounds 247-342 was measured at 30 min. + indicates binding
activity from 5% to 25% ++ indicates binding activity greater than
25% and up to 50% +++ indicates binding activity greater than 50%
and up to 75% ++++ indicates binding activity greater than 75%
TABLE-US-00006 TABLE 5 Activity of Representative Compounds of
Structure (II)* Binding No. % No. Binding % No. Binding % No.
Binding % II-1 ++++ II-2 +++ II-3 ++++ II-4 +++ II-5 + II-6 + II-7
++ II-8 ++ II-9 + II-10 + II-11 ++++ II-12 +++ II-13 ++ II-14 +++
II-15 + II-16 + II-17 ++ II-18 ++++ II-19 +++ II-20 ++ II-21 +
II-22 + II-23 + II-24 + II-25 + II-26 + II-27 ++ II-28 +++ II-29 +
II-30 + II-31 ++++ II-32 ++++ II-33 + II-34 + II-35 ++++ II-36 ++++
II-37 ++ II-38 + II-39 +++ II-40 ++++ II-41 ++++ II-42 + II-43 ++++
II-44 + II-45 ++++ II-46 + II-47 + II-48 ++ II-49 ++++ N/A N/A N/A
N/A N/A N/A *Binding activity determined at 24 hrs. + indicates
binding activity from 5% to 15% ++ indicates binding activity
greater than 15% and up to 25% +++ indicates binding activity
greater than 25% and up to 50% ++++ indicates binding activity
greater than 50%
TABLE-US-00007 TABLE 6 Activity of Representative Compounds of
Structure (II)* Binding No. % No. Binding % No. Binding % No.
Binding % II-50 ++ II-51 ++ II-52 + II-53 ++ II-54 ++ II-55 ++
II-56 ++ II-57 ++ II-58 + II-59 ++ II-60 ++ II-61 + II-62 + II-63
++ II-64 ++ II-65 + II-66 + II-67 ++ II-68 ++ II-69 ++ II-70 ++
II-71 + II-72 + II-73 + II-74 + II-75 + II-76 N/A N/A N/A *Binding
activity determined at 2 hrs. + indicates binding activity from 5%
to 20% ++ indicates binding activity greater than 20%
TABLE-US-00008 TABLE 7 Activity of Representative Compounds of
Structure (III)* Binding No. % No. Binding % No. Binding % No.
Binding % III-1 + III-2 ++ III-3 ++++ III-4 ++ III-5 ++++ III-6 ++
III-7 +++ III-8 + III-9 + III-10 + III-11 + III-12 +++ III-13 +
III-14 + III-15 + III-16 + III-17 ++++ III-18 + III-19 + III-20 +
III-21 ++ III-22 + III-23 +++ III-24 ++++ III-25 +++ III-26 ++
III-27 +++ III-28 ++ III-29 +++ III-30 ++ III-31 + III-32 ++++
III-33 + III-34 +++ III-35 ++ III-36 + *Binding activity determined
at 24 hrs. + indicates binding activity from 5% to 10% ++ indicates
binding activity greater than 10% and up to 20% +++ indicates
binding activity greater than 20% and up to 30% ++++ indicates
binding activity greater than 30%
TABLE-US-00009 TABLE 8 Activity of Representative Compounds of
Structure (III)* Binding No. % No. Binding % No. Binding % No.
Binding % III-37 ++ III-38 ++ III-39 ++ III-40 + III-41 + III-42 +
III-43 + N/A N/A *Binding activity determined at 2 hrs. + indicates
binding activity from 5% to 20% ++ indicates binding activity
greater than 20%
[0987] All of the U.S. patents, U.S. patent application
publications, U.S. patent applications, foreign patents, foreign
patent applications and non-patent publications referred to in this
specification or the attached Application Data Sheet are
incorporated herein by reference, in their entirety to the extent
not inconsistent with the present description.
[0988] From the foregoing it will be appreciated that, although
specific embodiments of the invention have been described herein
for purposes of illustration, various modifications may be made
without deviating from the spirit and scope of the invention.
Accordingly, the invention is not limited except as by the appended
claims.
* * * * *