U.S. patent application number 15/758982 was filed with the patent office on 2020-06-04 for inhibitors of cyclin-dependent kinases.
This patent application is currently assigned to Dana-Farber Cancer Institute, Inc.. The applicant listed for this patent is Dana-Farber Cancer Institute, Inc.. Invention is credited to Nathanael S. Gray, Mingfeng Hao, Nicholas Paul Kwiatkowski, Tinghu Zhandg.
Application Number | 20200172499 15/758982 |
Document ID | / |
Family ID | 58240329 |
Filed Date | 2020-06-04 |
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United States Patent
Application |
20200172499 |
Kind Code |
A9 |
Gray; Nathanael S. ; et
al. |
June 4, 2020 |
INHIBITORS OF CYCLIN-DEPENDENT KINASES
Abstract
The present invention provides novel compounds of Formulae (I'),
(I), (II'), and (II), and pharmaceutically acceptable salts,
solvates, hydrates, polymorphs, co-crystals, tautomers,
stereoisomers, isotopically labeled derivatives, prodrugs, and
compositions thereof. Also provided are methods and kits involving
the inventive compounds or compositions for treating and/or
preventing proliferative diseases (e.g., cancers (e.g., leukemia,
acute lymphoblastic leukemia, lymphoma, Burkitt's lymphoma,
melanoma, multiple myeloma, breast cancer, Ewing's sarcoma,
osteosarcoma, brain cancer, ovarian cancer, neuroblastoma, lung
cancer, colorectal cancer), benign neoplasms, diseases associated
with angiogenesis, inflammatory diseases, autoinflammatory
diseases, and autoimmune diseases) in a subject. Treatment of a
subject with a proliferative disease using a compound or
composition of the invention may inhibit the aberrant activity of a
kinase, such as a cyclin-dependent kinase (CDK) (e.g., CDK7, CDK12,
or CDK13), and therefore, induce cellular apoptosis and/or inhibit
transcription in the subject.
Inventors: |
Gray; Nathanael S.;
(Boston,, MA) ; Zhandg; Tinghu; (Brookline,
MA) ; Kwiatkowski; Nicholas Paul; (Brookline, MA)
; Hao; Mingfeng; (Boston, MA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Dana-Farber Cancer Institute, Inc. |
Boston |
MA |
US |
|
|
Assignee: |
Dana-Farber Cancer Institute,
Inc.
Boston
MA
|
Prior
Publication: |
|
Document Identifier |
Publication Date |
|
US 20180362483 A1 |
December 20, 2018 |
|
|
Family ID: |
58240329 |
Appl. No.: |
15/758982 |
Filed: |
September 9, 2016 |
PCT Filed: |
September 9, 2016 |
PCT NO: |
PCT/US16/51118 PCKC 00 |
371 Date: |
March 9, 2018 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
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62216271 |
Sep 9, 2015 |
|
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61P 35/00 20180101;
A61K 31/497 20130101; C07D 417/04 20130101; C07D 417/12 20130101;
A61K 31/497 20130101; C07D 277/46 20130101; A61P 25/00 20180101;
A61K 2300/00 20130101; C07D 417/14 20130101 |
International
Class: |
C07D 277/46 20060101
C07D277/46; C07D 417/04 20060101 C07D417/04; C07D 417/14 20060101
C07D417/14; C07D 417/12 20060101 C07D417/12 |
Goverment Interests
GOVERNMENT SUPPORT
[0002] This invention was made with government support under grant
number 1 R01 CA179483-01A1 awarded by the National Institutes of
Health. The government has certain rights in the invention.
Claims
1. A compound of Formula (I'): ##STR00370## or a pharmaceutically
acceptable salt thereof, wherein: R.sup.1 is hydrogen, halogen, or
optionally substituted alkyl; M is O, S, or NR.sup.M; R.sup.M is
hydrogen, optionally substituted alkyl, optionally substituted
alkenyl, optionally substituted alkynyl, optionally substituted
carbocyclyl, optionally substituted heterocyclyl, optionally
substituted aryl, optionally substituted heteroaryl, optionally
substituted acyl, or a nitrogen protecting group; Ring A is
optionally substituted monocyclic carbocyclyl, optionally
substituted monocyclic heterocyclyl, optionally substituted phenyl,
or optionally substituted monocyclic heteroaryl; Ring B is
optionally substituted monocyclic carbocyclyl, optionally
substituted monocyclic heterocyclyl, optionally substituted phenyl,
or optionally substituted monocyclic heteroaryl; Ring C is
optionally substituted monocyclic carbocyclyl, optionally
substituted monocyclic heterocyclyl, optionally substituted
monocyclic or bicyclic aryl, or optionally substituted monocyclic
or bicyclic heteroaryl; each instance of R.sup.A, R.sup.B, and
R.sup.C is independently hydrogen, halogen, optionally substituted
alkyl, optionally substituted alkenyl, optionally substituted
alkynyl, optionally substituted carbocyclyl, optionally substituted
heterocyclyl, optionally substituted aryl, optionally substituted
heteroaryl, --OR.sup.a, --N(R.sup.a).sub.2, --SR.sup.a, --CN,
--SCN, --NO.sub.2, --N.sub.3, or optionally substituted acyl; each
instance of R.sup.a is independently hydrogen, optionally
substituted alkyl, optionally substituted alkenyl, optionally
substituted alkynyl, optionally substituted carbocyclyl, optionally
substituted heterocyclyl, optionally substituted aryl, optionally
substituted heteroaryl, optionally substituted acyl, a nitrogen
protecting group when attached to a nitrogen atom, an oxygen
protecting group when attached to an oxygen atom, or a sulfur
protecting group when attached to a sulfur atom, or two instances
of R.sup.a are joined to form a substituted or unsubstituted,
heterocyclic ring, or substituted or unsubstituted, heteroaryl
ring; each of a1, b1, and c1 is independently 0, 1, 2, 3, 4, 5, or
6, as valency permits; L.sup.1 is --CH.sub.2--,
.sup.lc--S(.dbd.O).sub.2--.sup.lb, --O--, --S--, --NR.sup.L1--,
--C(.dbd.O)--, .sup.lc--NR.sup.L1C(.dbd.O)--.sup.lb,
.sup.lc--C(.dbd.O)NR.sup.L1--.sup.lb, .sup.lc--OC(.dbd.O).sup.lb,
or .sup.lc--C(.dbd.O)O--.sup.lb; wherein .sup.lb indicates the
point of attachment is to Ring B; and .sup.lc indicates the point
of attachment is to Ring C; L.sup.2 is --O--, --S--, --NR.sup.L2--,
.sup.lb--NR.sup.L2C(.dbd.O)--.sup.lm,
.sup.lb--C(.dbd.O)NR.sup.L2--.sup.lm; wherein .sup.lb indicates the
point of attachment is to Ring B; and .sup.lm indicates the point
of attachment is to the heteroaryl ring with M; X is
.sup.xm--CH.sub.2CH.sub.2--.sup.xa, .sup.xm--CH.dbd.CH--.sup.xa,
.sup.xm--CH.sub.2--NR.sup.LXxa,
.sup.xm--CH.sub.2--O--CH.sub.2--.sup.xa,
.sup.xm--CH.sub.2--NR.sup.LX--CH.sub.2--.sup.xa, --O--, --S--,
--NR.sup.LX--, .sup.xm--O--CH.sub.2--.sup.xa,
.sup.xm--S--CH.sub.2--.sup.xa,
.sup.xm--S--C(.dbd.O)CH.sub.2--.sup.xa, or
.sup.xm--NR.sup.LX--CH.sub.2--.sup.xa; wherein .sup.xa indicates
the point of attachment is to Ring A; and .sup.xm indicates the
point of attachment is to the heteroaryl ring with M; each of
R.sup.L1, R.sup.L2, and R.sup.LX is independently hydrogen,
optionally substituted C.sub.1-6 alkyl, or a nitrogen protecting
group; R.sup.2 is any of Formulae (i-1)-(i-41): ##STR00371##
##STR00372## ##STR00373## ##STR00374## ##STR00375## ##STR00376##
wherein: L.sup.3 is a bond or an optionally substituted C.sub.1-4
hydrocarbon chain, optionally wherein one or more carbon units of
the hydrocarbon chain are independently replaced with --C.dbd.O--,
--O--, --S--, --NR.sup.L3a--, --NR.sup.L3aC(.dbd.O)--,
--C(.dbd.O)NR.sup.L3a--, --SC(.dbd.O)--, --C(.dbd.O)S--,
--OC(.dbd.O)--, --C(.dbd.O)O--, --NR.sup.L3aC(.dbd.S)--,
--C(.dbd.S)NR.sup.L3a--, trans-CR.sup.L3b.dbd.CR.sup.L3b--,
cis-CR.sup.L3b.dbd.CR.sup.L3b--, --C.ident.C--, --S(.dbd.O)--,
--S(.dbd.O)O--, --OS(.dbd.O)--, --S(.dbd.O)NR.sup.L3a--,
--NR.sup.L3aS(.dbd.O)--, --S(.dbd.O).sub.2--, --S(.dbd.O).sub.2O--,
--OS(.dbd.O).sub.2--, --S(.dbd.O).sub.2NR.sup.L3a--, or
--NR.sup.L3aS(.dbd.O).sub.2--, wherein R.sup.L3a is hydrogen,
optionally substituted C.sub.1-6 alkyl, or a nitrogen protecting
group, and wherein each occurrence of R.sup.L3b is independently
hydrogen, halogen, optionally substituted alkyl, optionally
substituted alkenyl, optionally substituted alkynyl, optionally
substituted carbocyclyl, optionally substituted heterocyclyl,
optionally substituted aryl, or optionally substituted heteroaryl,
or two R.sup.L3b groups are joined to form an optionally
substituted carbocyclic or optionally substituted heterocyclic
ring; L.sup.4 is a bond or an optionally substituted, branched or
unbranched C.sub.1-6 hydrocarbon chain; each of R.sup.E1, R.sup.E2,
and R.sup.E3 is independently hydrogen, halogen, optionally
substituted alkyl, optionally substituted alkenyl, optionally
substituted alkynyl, optionally substituted carbocyclyl, optionally
substituted heterocyclyl, optionally substituted aryl, optionally
substituted heteroaryl, --CN, --CH.sub.2OR.sup.EE,
--CH.sub.2N(R.sup.EE).sub.2, --CH.sub.2SR.sup.EE, --OR.sup.EE,
--N(R.sup.EE).sub.2, --Si(R.sup.EE).sub.3, and --SR.sup.EE, wherein
each occurrence of R.sup.EE is independently hydrogen, optionally
substituted alkyl, optionally substituted alkoxy, optionally
substituted alkenyl, optionally substituted alkynyl, optionally
substituted carbocyclyl, optionally substituted heterocyclyl,
optionally substituted aryl, or optionally substituted heteroaryl,
or two R.sup.EE groups are joined to form an optionally substituted
heterocyclic ring; or R.sup.E1 and R.sup.E3, or R.sup.E2 and
R.sup.E3, or R.sup.E1 and R.sup.E2 are joined to form an optionally
substituted carbocyclic or optionally substituted heterocyclic
ring; R.sup.E4 is a leaving group; R.sup.E5 is halogen; R.sup.E6 is
hydrogen, optionally substituted C.sub.1-6 alkyl, or a nitrogen
protecting group; each instance of Y is independently O, S, or
NR.sup.E7, wherein R.sup.E7 is hydrogen, optionally substituted
C.sub.1-6 alkyl, or a nitrogen protecting group; a is 1 or 2; and
each instance of z is independently 0, 1, 2, 3, 4, 5, or 6, as
valency permits.
2. (canceled)
3. The compound of claim 1, wherein the compound is of the formula:
##STR00377## or a pharmaceutically acceptable salt thereof.
4. The compound of claim 1, or a pharmaceutically acceptable salt
thereof, wherein the Ring B is optionally substituted monocyclic
heterocyclyl.
5. The compound of claim 1, or a pharmaceutically acceptable salt
thereof, wherein the Ring B is optionally substituted
piperidinyl.
6. The compound of claim 5, wherein the compound is of Formula
(I-i), (I-ii), (I-ii-a), (I-ii-b), (I-ii-c), (I-iii), or (I'-i):
##STR00378## ##STR00379## or a pharmaceutically acceptable salt
thereof.
7-11. (canceled)
12. The compound of claim 1, or a pharmaceutically acceptable salt
thereof, wherein Ring B is optionally substituted monocyclic
carbocyclyl.
13. (canceled)
14. The compound of claim 1, wherein the compound is of Formula
(I-iv), (I-iv-a), or (I-iv-b): ##STR00380## or a pharmaceutically
acceptable salt thereof.
15-17. (canceled)
18. The compound of claim 1, or a pharmaceutically acceptable salt
thereof, wherein Ring C is optionally substituted.bicyclic aryl or
heteroaryl.
19-20. (canceled)
21. A compound of Formula (II'): ##STR00381## or a pharmaceutically
acceptable salt thereof, wherein: R.sup.1 is hydrogen, halogen, or
optionally substituted alkyl; M is O, S, or NR.sup.M; R.sup.M is
hydrogen, optionally substituted alkyl, optionally substituted
alkenyl, optionally substituted alkynyl, optionally substituted
carbocyclyl, optionally substituted heterocyclyl, optionally
substituted aryl, optionally substituted heteroaryl, optionally
substituted acyl, or a nitrogen protecting group; Ring A is
optionally substituted monocyclic carbocyclyl, optionally
substituted monocyclic heterocyclyl, optionally substituted phenyl,
or optionally substituted monocyclic heteroaryl; Ring B is
optionally substituted monocyclic carbocyclyl, optionally
substituted monocyclic heterocyclyl, optionally substituted phenyl,
or optionally substituted monocyclic heteroaryl; each instance of
R.sup.A and R.sup.B is independently hydrogen, halogen, optionally
substituted alkyl, optionally substituted alkenyl, optionally
substituted alkynyl, optionally substituted carbocyclyl, optionally
substituted heterocyclyl, optionally substituted aryl, optionally
substituted heteroaryl, --OR.sup.a, --N(R.sup.a).sub.2, --SR.sup.a,
--CN, --SCN, --C(.dbd.NR.sup.a)R.sup.a, --C(.dbd.NR.sup.a)OR.sup.a,
--C(.dbd.NR.sup.a)N(R.sup.a).sub.2, --C(.dbd.O)R.sup.a,
--C(.dbd.O)OR.sup.a, --C(.dbd.O)N(R.sup.a).sub.2, --NO.sub.2,
--NR.sup.aC(.dbd.O)R.sup.a, --NR.sup.aC(.dbd.O)OR.sup.a,
--NR.sup.aC(.dbd.O)N(R.sup.a).sub.2, --OC(.dbd.O)R.sup.a,
--OC(.dbd.O)OR.sup.a, or --OC(.dbd.O)N(R.sup.a).sub.2; each
instance of R.sup.a is independently hydrogen, optionally
substituted acyl, optionally substituted alkyl, optionally
substituted alkenyl, optionally substituted alkynyl, optionally
substituted carbocyclyl, optionally substituted heterocyclyl,
optionally substituted aryl, optionally substituted heteroaryl, a
nitrogen protecting group when attached to a nitrogen atom, an
oxygen protecting group when attached to an oxygen atom, or a
sulfur protecting group when attached to a sulfur atom, or two
instances of R.sup.a are joined to form a substituted or
unsubstituted, heterocyclic ring, or substituted or unsubstituted,
heteroaryl ring; each of a1 and b1 is independently 0, 1, 2, 3, 4,
5, or 6, as valency permits; L.sup.2 is --O--, --S--,
--NR.sup.L2--, .sup.lb--NR.sup.L2C(.dbd.O)--.sup.lm,
.sup.lb--C(.dbd.O)NR.sup.L2--.sup.lm; wherein .sup.lb indicates the
point of attachment is to Ring B; and .sup.lm indicates the point
of attachment is to the heteroaryl ring with M; X is a bond, --O--,
--S--, --NR.sup.LX--, .sup.xm--O--CH.sub.2--.sup.xa,
.sup.xm--S--CH.sub.2--.sup.xa, or
.sup.xm--NR.sup.LX--CH.sub.2--.sup.xa; wherein .sup.xa indicates
the point of attachment is to Ring A; and .sup.xm indicates the
point of attachment is to the heteroaryl ring with M; each of
R.sup.L2 and R.sup.LX is independently hydrogen, optionally
substituted C.sub.1-6 alkyl, or a nitrogen protecting group;
R.sup.2 is any of Formulae (i-1)-(i-41): ##STR00382## ##STR00383##
##STR00384## ##STR00385## ##STR00386## ##STR00387## wherein:
L.sup.3 is a bond or an optionally substituted C.sub.1-4
hydrocarbon chain, optionally wherein one or more carbon units of
the hydrocarbon chain are independently replaced with --C.dbd.O--,
--O--, --S--, --NR.sup.L3a--, --NR.sup.L3aC(.dbd.O)--,
--C(.dbd.O)NR.sup.L3a--, --SC(.dbd.O)--, --C(.dbd.O)S--,
--OC(.dbd.O)--, --C(.dbd.O)O--, --NR.sup.L3aC(.dbd.S)--,
--C(.dbd.S)NR.sup.L3a--, trans-CR.sup.L3b.dbd.CR.sup.L3b--,
cis-CR.sup.L3b.dbd.CR.sup.L3b--, --C.ident.C--, --S(.dbd.O)--,
--S(.dbd.O)O--, --OS(--O)--, --S(--O)NR.sup.L3a--,
--NR.sup.L3aS(.dbd.O)--, --S(.dbd.O).sub.2--, --S(.dbd.O).sub.2O--,
--OS(.dbd.O).sub.2--, --S(.dbd.O).sub.2NR.sup.L3a--, or
--NR.sup.L3aS(.dbd.O).sub.2--, wherein R.sup.L3a is hydrogen,
optionally substituted C.sub.1-6 alkyl, or a nitrogen protecting
group, and wherein each occurrence of R.sup.L3b is independently
hydrogen, halogen, optionally substituted alkyl, optionally
substituted alkenyl, optionally substituted alkynyl, optionally
substituted carbocyclyl, optionally substituted heterocyclyl,
optionally substituted aryl, or optionally substituted heteroaryl,
or two R.sup.L3b groups are joined to form an optionally
substituted carbocyclic or optionally substituted heterocyclic
ring; L.sup.4 is a bond or an optionally substituted, branched or
unbranched C.sub.1-6 hydrocarbon chain; each of R.sup.E1, R.sup.E2,
and R.sup.E3 is independently hydrogen, halogen, optionally
substituted alkyl, optionally substituted alkenyl, optionally
substituted alkynyl, optionally substituted carbocyclyl, optionally
substituted heterocyclyl, optionally substituted aryl, optionally
substituted heteroaryl, --CN, --CH.sub.2OR.sup.EE,
--CH.sub.2N(R.sup.EE).sub.2, --CH.sub.2SR.sup.EE, --OR.sup.EE,
--N(R.sup.EE).sub.2, --Si(R.sup.EE).sub.3, and --SR.sup.EE, wherein
each occurrence of R.sup.EE is independently hydrogen, optionally
substituted alkyl, optionally substituted alkoxy, optionally
substituted alkenyl, optionally substituted alkynyl, optionally
substituted carbocyclyl, optionally substituted heterocyclyl,
optionally substituted aryl, or optionally substituted heteroaryl,
or two R.sup.EE groups are joined to form an optionally substituted
heterocyclic ring; or R.sup.E1 and R.sup.E3, or R.sup.E2 and
R.sup.E3, or R.sup.E1 and R.sup.E2 are joined to form an optionally
substituted carbocyclic or optionally substituted heterocyclic
ring; R.sup.E4 is a leaving group; R.sup.E5 is halogen; R.sup.E6 is
hydrogen, optionally substituted C.sub.1-6 alkyl, or a nitrogen
protecting group; each instance of Y is independently O, S, or
NR.sup.E7, wherein R.sup.E7 is hydrogen, optionally substituted
C.sub.1-6 alkyl, or a nitrogen protecting group; a is 1 or 2; and
each instance of z is independently 0, 1, 2, 3, 4, 5, or 6, as
valency permits.
22-30. (canceled)
31. The compound of claim 1, or a pharmaceutically acceptable salt
thereof, wherein Ring A is optionally substituted monocyclic
heteroaryl.
32-39. (canceled)
40. The compound of claim 1, or a pharmaceutically acceptable salt
thereof, wherein Ring A is optionally substituted phenyl.
41-46. (canceled)
47. The compound of claim 1, or a pharmaceutically acceptable salt
thereof, wherein Ring A is optionally substituted monocyclic
carbocyclyl.
48. (canceled)
49. The compound of claim 1, or a pharmaceutically acceptable salt
thereof, wherein Ring A is optionally substituted monocyclic
heterocyclyl.
50-54. (canceled)
55. The compound of claim 1, or a pharmaceutically acceptable salt
thereof, wherein Ring A is optionally substituted 6-membered
heteroaryl.
56. (canceled)
57. The compound of claim 1, or a pharmaceutically acceptable salt
thereof, wherein L.sup.1 is --CH.sub.2-- or
.sup.lc--S(.dbd.O).sub.2--.sup.lb.
58-82. (canceled)
83. The compound of claim 1, or a pharmaceutically acceptable salt
thereof, wherein R.sup.2 is of Formula (i-1): ##STR00388##
84-90. (canceled)
91. The compound of claim 1, wherein the compound is of one of the
following formulae: ##STR00389## ##STR00390## ##STR00391##
##STR00392## ##STR00393## or a pharmaceutically acceptable salt
thereof.
92. The compound of claim 1, wherein the compound is of one of the
following formulae: ##STR00394## ##STR00395## ##STR00396##
##STR00397## ##STR00398## ##STR00399## ##STR00400## ##STR00401##
##STR00402## ##STR00403## ##STR00404## ##STR00405## ##STR00406##
##STR00407## ##STR00408## ##STR00409## ##STR00410## or a
pharmaceutically acceptable salt thereof.
93. The compound of claim 1, wherein the compound is of one of the
following formulae: ##STR00411## ##STR00412## ##STR00413##
##STR00414## ##STR00415## ##STR00416## ##STR00417## or a
pharmaceutically acceptable salt thereof.
94-95. (canceled)
96. A pharmaceutical composition comprising a compound of claim 1,
or a pharmaceutically acceptable salt thereof, and optionally a
pharmaceutically acceptable excipient.
97. (canceled)
98. A method of treating a proliferative disease in a subject in
need thereof, the method comprising administering to the subject a
therapeutically effective amount of a compound of claim 1, or a
pharmaceutically acceptable salt thereof.
99-131. (canceled)
132. A method of inhibiting the activity of a cyclin-dependent
kinase (CDK) in a biological sample or subject, the method
comprising administering to the subject or contacting the
biological sample with a therapeutically effective amount of a
compound of claim 1, or a pharmaceutically acceptable salt
thereof.
133-153. (canceled)
Description
RELATED APPLICATIONS
[0001] This application claims priority under 35 U.S.C. .sctn.
119(e) to U.S. Provisional application, U.S. Ser. No. 62/216,271,
filed Sep. 9, 2015, which is incorporated herein by reference.
BACKGROUND OF THE INVENTION
[0003] The members of the cyclin-dependent kinase (CDK) family play
critical regulatory roles in cell proliferation. There are
currently twenty known mammalian CDKs. While CDK7 to CDK13 have
been linked to transcription, CDK1, 2, 4, and 6 show demonstrable
association with the cell cycle.
[0004] Unique among the mammalian CDKs, CDK7 has consolidated
kinase activities, regulating both the cell cycle and
transcription. In the cytosol, CDK7 exists as a heterotrimeric
complex and is believed to function as a CDK1/2-activating kinase
(CAK), whereby phosphorylation of conserved residues in CDK1/2 by
CDK7 is required for full catalytic CDK activity and cell cycle
progression (Desai et al., "Effects of phosphorylation by CAK on
cyclin binding by CDC2 and CDK2." Mol. Cell Biol. 15, 345-350
(1995); Kaldis et al., "Analysis of CAK activities from human
cells." Eur. J. Biochem. 267, 4213-4221 (2000); Larochelle et al.,
"Requirements for CDK7 in the assembly of CDK1/cyclin B and
activation of CDK2 revealed by chemical genetics in human cells."
Mol. Cell 25, 839-850 (2007)). In the nucleus, CDK7 forms the
kinase core of the RNA polymerase (RNAP) II general transcription
factor complex and is charged with phosphorylating the C-terminal
domain (CTD) of RNAP II, a requisite step in gene transcriptional
initiation (Serizawa. et al., "Association of CDK-activating kinase
subunits with transcription factor TFIIH." Nature 374, 280-282
(1995); Shiekhattar et al., "CDK-activating kinase complex is a
component of human transcription factor TFIIH." Nature 374, 283-287
(1995); Drapkin et al., "Human cyclin-dependent kinase-activating
kinase exists in three distinct complexes." Proc. Natl. Acad. Sci.
U.S.A. 93, 6488-6493 (1996); Liu. et al., "Two cyclin-dependent
kinases promote RNA polymerase II transcription and formation of
the scaffold complex." Mol. Cell Biol. 24, 1721-1735 (2004); Akhtar
et al., "TFIIH kinase places bivalent marks on the carboxy-terminal
domain of RNA polymerase II." Mol. Cell 34, 387-393 (2009);
Glover-Cutter et al., "TFIIH-associated CDK7 kinase functions in
phosphorylation of C-terminal domain Ser7 residues,
promoter-proximal pausing, and termination by RNA polymerase II."
Mol. Cell Biol. 29, 5455-5464 (2009)). Together, the two functions
of CDK7, i.e., CAK and CTD phosphorylation, support critical facets
of cellular proliferation, cell cycling, and transcription.
[0005] CDK12 and CDK13 were identified in cDNA screens for cell
cycle regulators. Because their cyclin partners were not yet known,
they were initially named CRKRS and CDC2L5 (Ko et al., J. Cell
Sci., 2001, 114, 2591-2603; Marques et al., Biochem Biophys Res
Commun., 2000, 279(3):832-837), respectively. They were found to be
1490- and 1512-amino acid proteins, respectively, with a conserved
central CTD kinase domain and degenerate RS domains identified in
their N- and C-terminal regions (Even et al., J Cell Biochem.,
2006, 99(3), 890-904).
[0006] Evidence has shown CDK12 and CDK13 play an important role in
cancer development. A comprehensive genomic approach identified
CDK12 to be one of the most frequently somatically mutated genes in
high-grade serous ovarian cancer, the most fatal form of the
disease (Erratum, Nature, 2011, 474(7353), 609-615). Several
identified point mutations in the kinase domain point to the
critical importance of the kinase activity of CDK12 for the
development/progression of this disease. CDK12 has also been found
to contribute to the development of breast cancer. Notably, CDK12
is located on chromosome 17, within the 17q21 locus that contains
several candidate genes for breast cancer susceptibility
(Kauraniemi et al., Cancer Res., 2001, 61(22), 8235-8240), and it
is co-amplified with the tyrosine kinase receptor ERBB2, a protein
amplified and overexpressed in about 20% of breast tumors. Gene
fusion between CDK12 and ERBB2 was also detected in gastric cancer
(Zang et al., Cancer Res., 2011, 71(1), 29-39). CDK12 is also
implicated in the modification of tamoxifen sensitivity in
estrogen-positive breast cancer via the modulation of the
mitogen-activated protein kinase pathway (Iorns et al.,
Carcinogenesis, 2009, 30(10):1696-1701).
[0007] Due to the important regulatory functions of kinases, such
as CDK7, CDK12, and CDK13, in cell cycle control, cell
proliferation, differentiation, and apoptosis, it is important to
develop modulators of the activities of these kinases, including
selective modulators, for use as research tools as well as
therapeutic agents in the treatment of diseases.
SUMMARY OF THE INVENTION
[0008] Cyclin dependent kinases (CDKs), e.g., CDK7, CDK12, and
CDK13, are key regulators of the cell cycle. Their successive
activation and inactivation drives the cycle forward. The activity
of CDKs is regulated by multiple mechanisms such as positive and
negative phosphorylation, binding of regulatory proteins like
cyclins, and CDK inhibitors. Most CDKs require the phosphorylation
of a threonine residue located in the T-loop to achieve full kinase
activity. This threonine residue is conserved in all CDKs that
function in cell cycle regulation. The enzyme responsible for this
phosphorylation is therefore termed CDK-activating-kinase or CAK.
CAK complexes have been found to be composed of CDK7, CDK12, CDK13,
cyclin H, and MAT1. Besides its CAK function, CDK7, CDK12, and
CDK13 also play a role in transcription and possibly in DNA repair.
This suggests that the CDK7, CDK12, and CDK13 enzyme complexes are
involved in multiple functions in the cell, e.g., cell cycle
control, apoptosis, transcription regulation, and DNA repair.
[0009] The present invention provides compounds of Formulae (I'),
(II'), (I), (II), and pharmaceutically acceptable salts, solvates,
hydrates, polymorphs, co-crystals, tautomers, stereoisomers,
isotopically labeled derivatives, prodrugs, and compositions
thereof. The compounds of Formulae (I'), (II'), (I), (II), and
pharmaceutically acceptable salts, solvates, hydrates, polymorphs,
co-crystals, tautomers, stereoisomers, isotopically labeled
derivatives, prodrugs, and compositions thereof, may inhibit the
activity of a kinase. The compounds described herein may in certain
embodiments selectively inhibit specific CDK subtypes, for example,
CDK7, CDK12, or CDK13. In certain embodiments, the compounds of
Formulae (I'), (II'), (I), and (II) are selective for CDK7 compared
to other kinases. In certain embodiments, the compounds of Formulae
(I'), (II'), (I), and (II) are selective for CDK12 and/or CDK13
compared to other kinases. The present invention also provides
methods of using the inventive compounds, and pharmaceutically
acceptable salts, solvates, hydrates, polymorphs, co-crystals,
tautomers, stereoisomers, isotopically labeled derivatives,
prodrugs, and compositions thereof, to study the inhibition of a
kinase (e.g., CDK7, CDK12, and/or CDK13) and as therapeutics for
the prevention and/or treatment of diseases associated with the
overexpression and/or aberrant activity of a kinase (e.g., CDK7,
CDK12, and/or CDK13). In certain embodiments, the inventive
compounds are used for the prevention and/or treatment of
proliferative diseases (e.g., cancers (e.g., leukemia, acute
lymphoblastic leukemia, lymphoma, Burkitt's lymphoma, melanoma,
multiple myeloma, breast cancer, Ewing's sarcoma, osteosarcoma,
brain cancer, neuroblastoma, lung cancer, colorectal cancer),
benign neoplasms, diseases associated with angiogenesis,
inflammatory diseases, autoinflammatory diseases, and autoimmune
diseases) in a subject.
[0010] Since the discovery of selective inhibitors of CDK7, CDK12,
and CDK13 has been hampered by the high sequence and structural
similarities of the kinase domain of CDK family members, the
development of selective inhibitors of the transcriptional
cyclin-dependent kinases (tCDKs) will allow dissection of their
individual contributions to the regulation of transcription and
evaluation of their therapeutic potential. Without wishing to be
bound by any particular theory, the inventive compounds'
selectivity for CDK7, CDK12, and/or CDK13 may be due to the
compounds' ability to covalently modify a specific cysteine residue
of these kinases (e.g., Cys312 of CDK7, Cys1039 of CDK12, Cys1017
of CDK13).
[0011] In one aspect, the present invention provides compounds of
Formula (I'):
##STR00001##
and pharmaceutically acceptable salts, solvates, hydrates,
polymorphs, co-crystals, tautomers, stereoisomers, isotopically
labeled derivatives, and prodrugs thereof, wherein R.sup.1,
R.sup.2, R.sup.A, R.sup.B, R.sup.C, Ring A, Ring B, Ring C,
L.sup.1, L.sup.2, X, a1, b1, and c1 are as defined herein.
[0012] In one aspect, the present invention provides compounds of
Formula (I):
##STR00002##
and pharmaceutically acceptable salts, solvates, hydrates,
polymorphs, co-crystals, tautomers, stereoisomers, isotopically
labeled derivatives, and prodrugs thereof, wherein R.sup.1,
R.sup.2, R.sup.A, R.sup.B, R.sup.C, Ring A, Ring B, Ring C,
L.sup.1, L.sup.2, X, a1, b1, and c1 are as defined herein.
[0013] In one aspect, the present invention provides compounds of
Formula (II'):
##STR00003##
and pharmaceutically acceptable salts, solvates, hydrates,
polymorphs, co-crystals, tautomers, stereoisomers, isotopically
labeled derivatives, and prodrugs thereof, wherein R.sup.1,
R.sup.2, R.sup.A, R.sup.B, Ring A, Ring B, L.sup.2, X, a1, and b1
are as defined herein.
[0014] In one aspect, the present invention provides compounds of
Formula (II):
##STR00004##
and pharmaceutically acceptable salts, solvates, hydrates,
polymorphs, co-crystals, tautomers, stereoisomers, isotopically
labeled derivatives, and prodrugs thereof, wherein R.sup.1,
R.sup.2, R.sup.A, R.sup.B, Ring A, Ring B, L.sup.2, X, a1, and b1
are as defined herein.
[0015] In another aspect, the present disclosure provides
pharmaceutical compositions including a compound described herein,
and optionally a pharmaceutically acceptable excipient. In certain
embodiments, the pharmaceutical compositions described herein
include a therapeutically or prophylactically effective amount of a
compound described herein. The pharmaceutical composition may be
useful for treating a proliferative disease in a subject in need
thereof, preventing a proliferative disease in a subject in need
thereof, inhibiting the activity of a protein kinase in a subject,
biological sample, tissue, or cell, and/or inducing apoptosis in a
cell. In certain embodiments, the proliferative disease is an
inflammatory disease. In certain embodiments, the inflammatory
disease is rheumatoid arthritis, Crohn's disease, or fibrosis.
[0016] In another aspect, the present invention provides methods
for treating and/or preventing a proliferative disease. Exemplary
proliferative diseases which may be treated include cancer, benign
neoplasms, diseases associated with angiogenesis, inflammatory
diseases, autoinflammatory diseases, and autoimmune diseases. In
certain embodiments, the cancer is selected from the group
consisting of pancreatic cancer, lung cancer (e.g., small cell lung
cancer (SCLC), and non-small cell lung cancer), prostate cancer,
breast cancer, ovarian cancer, kidney cancer, liver cancer, Ewing's
sarcoma, osteosarcoma, brain cancer, neuroblastoma, and colorectal
cancer.
[0017] Another aspect of the invention relates to methods of
inhibiting the activity of a kinase (e.g., CDK (e.g., CDK7, CDK12,
CDK13)) using a compound described herein in a biological sample or
subject. In certain embodiments, the method involves the selective
inhibition of CDK7. In certain embodiments, the method involves the
selective inhibition of CDK12. In certain embodiments, the method
involves the selective inhibition of CDK13.
[0018] Also provided by the present invention are methods of
inhibiting the transcription of one or more genes in the cell of a
biological sample or subject using a compound described herein. The
transcription of genes affected by the activity of CDK7 may be
inhibited by a compound of the invention. In certain embodiments,
these genes are one or more selected from the group consisting of
MYC, RUNX1, MYB, TAL1, GATA3, KLF2, HNRPDL, p21, ASCL1, MYCN,
INSM1, NEUROD1, NEUROG1, FOXG1, FOXA1, SOX2, SOX4, BCL11A, OTX2,
GAT2, PHOX2B, PLK2, TAF1, CTGF, WEE1, SDIM, JUN, PIM1, IL8, and
FOS1. The transcription of genes affected by the activity of CDK12
may be inhibited by a compound of the invention. In certain
embodiments, these genes are one or more selected from the group
consisting of BRCA1, FANCI, ATR, FANCD2, APEX1, NEK9, CHEK1, CHEK2,
ATM, RAD51C, RAD51D, ORC3L, MDC1, TERF2, ERCC4, FANCF, PARP9,
RUNX1, MYB, TAL1, MCL1, MYC, BCL2, ETS1, and EWS-FLI. The
transcription of genes affected by the activity of CDK13 may be
inhibited by a compound of the invention. In certain embodiments,
the gene is SNORA38.
[0019] The present invention also provides methods of inhibiting
cell growth in a biological sample or subject. In still another
aspect, the present invention provides methods of inducing
apoptosis of a cell in a biological sample or subject.
[0020] The present invention provides methods for administering to
a subject in need thereof an effective amount of a compound, or
pharmaceutical composition thereof, as described herein. Also
described are methods for contacting a cell with an effective
amount of a compound, or pharmaceutical composition thereof, as
described herein. In certain embodiments, a method described herein
further includes administering to the subject an additional
pharmaceutical agent. In certain embodiments, a method described
herein further includes contacting the cell with an additional
pharmaceutical agent. The methods described herein may further
include performing radiotherapy, immunotherapy, and/or
transplantation on the subject.
[0021] In yet another aspect, the present invention provides
compounds of Formulae (I'), (II'), (I), (II), and pharmaceutically
acceptable salts, solvates, hydrates, polymorphs, co-crystals,
tautomers, stereoisomers, isotopically labeled derivatives,
prodrugs, and compositions thereof, for use in the treatment of a
disease (e.g., a proliferative disease such as cancer) in a
subject.
[0022] Another aspect of the present disclosure relates to kits
comprising a container with a compound, or pharmaceutical
composition thereof, as described herein. The kits described herein
may include a single dose or multiple doses of the compound or
pharmaceutical composition. The kits may be useful in a method of
the disclosure. In certain embodiments, the kit further includes
instructions for using the compound or pharmaceutical composition.
A kit described herein may also include information (e.g.
prescribing information) as required by a regulatory agency such as
the U.S. Food and Drug Administration (FDA).
[0023] The details of one or more embodiments of the invention are
set forth herein. Other features, objects, and advantages of the
invention will be apparent from the Detailed Description, Examples,
Figures, and Claims.
BRIEF DESCRIPTION OF THE DRAWINGS
[0024] The accompanying drawing, which are incorporated in and
constitute a part of this specification, illustrate several
embodiments of the invention and together with the description,
serve to explain the principles of the invention.
[0025] FIG. 1 shows the chemical structures of exemplary compounds
described herein and the IC.sub.50 values of the exemplary
compounds in inhibiting select cyclin-dependent kinases.
[0026] FIG. 2A shows that compound B12 exhibits binding of
intracellular CDK12 and CDK13 at concentrations between 62.5 nM to
1 .mu.M treatment. Jurkat cells treated with compound B12 for 6
hours show decreased pulldown of CDK12 and CDK13-associated cyclin
K by biotin-THZ1 relative to DMSO-treated cells. These data
indicate that B12 cellular treatment blocks biotin-THZ1 from
binding CDK12 and CDK13-associated cyclin K complexes by successful
binding of these complexes in cells at these concentrations. Cyclin
H (Cyc H) pulldown was not affected, indicating that CDK7 binding
is not affected. FIG. 2B shows the structure of compound B12.
[0027] FIG. 3 shows the binding of compounds B12, B15, and B16
(relative to DMSO control) to CDK12 and CDK13-associated cyclin K
at concentrations of 100 nM, 50 nM, 25 nM, and 12.5 nM. Jurkat
cells were treated with each compound (or DMSO) for 6 hours,
followed by lysis and pulldown with biotin-THZ1, and subsequent
western blotting for cyclin K (Cyc K) and cyclin H (Cyc H).
Compound B12 exhibits binding to CDK12 and CDK13-associated cyclin
K, while compounds B15 and B16 do not. Compound B12 is able to
block pulldown of CDK12 and CDK13-associated cyclin K down to 25 nM
treatment, while compounds B15 and B16 show very little effect on
cyclin K pulldown. These results indicate that at concentrations as
low as 25 nM B12 successfully targets intracellular CDK12 and
CDK13-associated cyclin K complexes.
[0028] FIG. 4 shows binding of intracellular CDK12 and
CDK13-associated cyclin K complexes by exemplified compounds.
Jurkat cells were treated with each compound at a concentration of
500 nM for 6 hours, followed by lysing and pulldown with
biotin-THZ1, and subsequent western blotting for cyclin K (Cyc K)
and cyclin H (Cyc H). Compounds B1, B3, B4, B5, B6, B8, B9, and B12
show a loss in CDK12 and CDK13-associated cyclin K pulldown,
indicating that these compounds successfully bind CDK12 and
CDK13-associated cyclin K complexes in cells thus blocking
biotin-THZ1 pull down, while compounds B5, B8, and B9 show a more
pronounced loss of cyclin H pulldown, indicating that these
compounds successfully target CDK7-cyclin H complexes in cells and
interfere with biotin-THZ1 pull down.
[0029] FIG. 5 shows exemplary results of growth assays of select
compounds described herein. Jurkat cells were plated at 30,000
cells/well and treated with a titration of compounds indicated.
Cells were allowed to grow for 72 hours. Cells were assayed using
CELLTITER GLO (Promega) to determine cell viability by measuring
the amount of ATP present, which is an indicator of cell metabolic
activity. Top panel: luminescent values (y-axis); concentration in
.mu.M (uM) (x-axis); the curves are generated using PRISM. Bottom
panel: IC.sub.50 values in .mu.M. Error bars indicate +/- standard
deviation. All compounds tested showed anti-proliferative effects
to varying extents.
[0030] FIG. 6 shows exemplary mass spectrum labeling of CDK12 with
compound B12. Compound B12 is able to label CDK12 once treated with
a 5-fold excess of compound B12 for 1 hour at 4.degree. C.
[0031] FIG. 7 shows the binding of intracellular CDK12 and
CDK13-associated cyclin K complexes by exemplified compounds.
Jurkat cells were treated with each compound at a concentration of
500 nM for 6 hours, followed by lysing and pull down with
biotin-THZ1, and subsequent western blotting for cyclin K (Cyc K)
and cyclin H (Cyc H). Compounds that decrease the pull down
efficiency of CDK12 and CDK13-associated cyclin K complexes
indicate those compounds successfully targeted intracellular CDK12
and CDK13-associated cyclin K complexes and therefore were able to
block biotin-THZ1 pull down of these complexes. In FIG. 7A,
compounds MFH 2-90-1 and MFH 2-102-1, show a loss in cyclin K
pulldown by biotin-THZ1, indicating that these compounds were able
to bind intracellular CDK12 and CDK13-associated cyclin K complexes
and block pull down by biotin-THZ1. In FIG. 7B, compounds MFH
2-90-1, MFH 3-103-1, and MFH 3-151-1, show a loss in cyclin K
pulldown by biotin-THZ1, indicating that these compounds were able
to bind intracellular CDK12 and CDK13-associated cyclin K complexes
and thus block pull down by biotin-THZ1.
[0032] FIG. 8 shows the binding of intracellular CDK12 and
CDK13-associated cyclin K complexes by exemplified compounds.
Jurkat cells were treated with each compound at a concentration of
500 nM for 6 hours, followed by lysing and pull down with
biotin-THZ1, and subsequent western blotting for cyclin K (Cyc K)
and cyclin H (Cyc H). Compounds that decrease the pull down
efficiency of CDK12 and CDK13-associated cyclin K complexes
indicate those compounds successfully targeted intracellular CDK12
and CDK13-associated cyclin K complexes and therefore were able to
block biotin-THZ1 pull down of these complexes. In FIG. 8A,
compound THZ 5-31 and MFH 2-90-1 show a loss in cyclin K pulldown
by biotin-THZ1, indicating that these compounds successfully
targeted CDK12 and CDK13-associated complexes in cells and block
biotin-THZ1 binding. In FIG. 8B, compounds THZ 5-31, MFH 2-90-1,
THZ-CE B-15, and THZ-CE B-16, show a loss in cyclin K pull down by
biotin-THZ1, indicating that these compounds successfully targeted
CDK12 and CDK13-associated complexes in cells and block biotin-THZ1
binding.
[0033] FIG. 9 shows the binding of intracellular CDK12 and
CDK13-associated cyclin K complexes by exemplified compounds.
Jurkat cells were treated with each compound at a concentration of
500 nM for 4 hours, followed by lysing and pull down with
biotin-THZ1, and subsequent western blotting for cyclin K (Cyc K)
and cyclin H (Cyc H). Compounds that decrease the pull down
efficiency of CDK12 and CDK13-associated cyclin K complexes
indicate those compounds successfully targeted intracellular CDK12
and CDK13-associated cyclin K complexes and therefore were able to
block biotin-THZ1 pull down of these complexes. In FIG. 9A,
compounds THZ 5-31, MFH 2-90-1, MFH 3-75-1, and MFH 3-81-1 show a
loss in cyclin K pull down by biotin-THZ1, indicating that these
compounds successfully targeted CDK12 and CDK13-associated
complexes in cells and block biotin-THZ1 binding. In FIG. 9B,
compounds THZ 5-31, MFH 4-70-1, and MFH 4-70-1 show a loss in
cyclin K pull down, by biotin-THZ1, indicating that these compounds
successfully targeted CDK12 and CDK13-associated complexes in cells
and block biotin-THZ1 binding.
DEFINITIONS
[0034] Definitions of specific functional groups and chemical terms
are described in more detail below. The chemical elements are
identified in accordance with the Periodic Table of the Elements,
CAS version, Handbook of Chemistry and Physics, 75.sup.th Ed.,
inside cover, and specific functional groups are generally defined
as described therein.
[0035] Additionally, general principles of organic chemistry, as
well as specific functional moieties and reactivity, are described
in Thomas Sorrell, Organic Chemistry, University Science Books,
Sausalito, 1999; Smith and March, March's Advanced Organic
Chemistry, 5.sup.th Edition, John Wiley & Sons, Inc., New York,
2001; Larock, Comprehensive Organic Transformations, VCH
Publishers, Inc., New York, 1989; and Carruthers, Some Modern
Methods of Organic Synthesis, 3.sup.rd Edition, Cambridge
University Press, Cambridge, 1987. The disclosure is not intended
to be limited in any manner by the exemplary listing of
substituents described herein.
[0036] Compounds described herein can comprise one or more
asymmetric centers, and thus can exist in various isomeric forms,
e.g., enantiomers and/or diastereomers. For example, the compounds
described herein can be in the form of an individual enantiomer,
diastereomer or geometric isomer, or can be in the form of a
mixture of stereoisomers, including racemic mixtures and mixtures
enriched in one or more stereoisomer. Isomers can be isolated from
mixtures by methods known to those skilled in the art, including
chiral high pressure liquid chromatography (HPLC) and the formation
and crystallization of chiral salts; or preferred isomers can be
prepared by asymmetric syntheses. See, for example, Jacques et al.,
Enantiomers, Racemates and Resolutions (Wiley Interscience, New
York, 1981); Wilen et al., Tetrahedron 33:2725 (1977); Eliel,
Stereochemistry of Carbon Compounds (McGraw-Hill, N Y, 1962); and
Wilen, Tables of Resolving Agents and Optical Resolutions p. 268
(E. L. Eliel, Ed., Univ. of Notre Dame Press, Notre Dame, Ind.
1972). The disclosure additionally encompasses compounds described
herein as individual isomers substantially free of other isomers,
and alternatively, as mixtures of various isomers.
[0037] When a range of values is listed, it is intended to
encompass each value and sub-range within the range. For example
"C.sub.1-6" is intended to encompass, C.sub.1, C.sub.2, C.sub.3,
C.sub.4, C.sub.5, C.sub.6, C.sub.1-6, C.sub.1-5, C.sub.1-4,
C.sub.1-3, C.sub.1-2, C.sub.2-6, C.sub.2-5, C.sub.2-4, C.sub.2-3,
C.sub.3-6, C.sub.3-5, C.sub.3-4, C.sub.4-6, C.sub.4-5, and
C.sub.5-6.
[0038] The term "aliphatic" includes both saturated and
unsaturated, straight chain (i.e., unbranched), branched, acyclic,
cyclic, or polycyclic aliphatic hydrocarbons, which are optionally
substituted with one or more functional groups. As will be
appreciated by one of ordinary skill in the art, "aliphatic" is
intended herein to include, but is not limited to, alkyl, alkenyl,
alkynyl, cycloalkyl, cycloalkenyl, and cycloalkynyl moieties. Thus,
the term "alkyl" includes straight, branched and cyclic alkyl
groups. An analogous convention applies to other generic terms such
as "alkenyl", "alkynyl", and the like. Furthermore, the terms
"alkyl", "alkenyl", "alkynyl", and the like encompass both
substituted and unsubstituted groups. In certain embodiments,
"lower alkyl" is used to indicate those alkyl groups (cyclic,
acyclic, substituted, unsubstituted, branched or unbranched) having
1-6 carbon atoms.
[0039] In certain embodiments, the alkyl, alkenyl, and alkynyl
groups employed in the disclosure contain 1-20 aliphatic carbon
atoms. In certain other embodiments, the alkyl, alkenyl, and
alkynyl groups employed in the disclosure contain 1-10 aliphatic
carbon atoms. In yet other embodiments, the alkyl, alkenyl, and
alkynyl groups employed in the disclosure contain 1-8 aliphatic
carbon atoms. In still other embodiments, the alkyl, alkenyl, and
alkynyl groups employed in the disclosure contain 1-6 aliphatic
carbon atoms. In yet other embodiments, the alkyl, alkenyl, and
alkynyl groups employed in the disclosure contain 1-4 carbon atoms.
Illustrative aliphatic groups thus include, but are not limited to,
for example, methyl, ethyl, n-propyl, isopropyl, cyclopropyl,
--CH.sub.2-cyclopropyl, vinyl, allyl, n-butyl, sec-butyl, isobutyl,
tert-butyl, cyclobutyl, --CH.sub.2-cyclobutyl, n-pentyl,
sec-pentyl, isopentyl, tert-pentyl, cyclopentyl,
--CH.sub.2-cyclopentyl, n-hexyl, sec-hexyl, cyclohexyl,
--CH.sub.2-cyclohexyl moieties and the like, which again, may bear
one or more substituents. Alkenyl groups include, but are not
limited to, for example, ethenyl, propenyl, butenyl,
1-methyl-2-buten-1-yl, and the like. Representative alkynyl groups
include, but are not limited to, ethynyl, 2-propynyl (propargyl),
1-propynyl, and the like.
[0040] The term "alkyl" refers to a radical of a straight-chain or
branched saturated hydrocarbon group having from 1 to 10 carbon
atoms ("C.sub.1-10 alkyl"). In some embodiments, an alkyl group has
1 to 9 carbon atoms ("C.sub.1-9 alkyl"). In some embodiments, an
alkyl group has 1 to 8 carbon atoms ("C.sub.1-8 alkyl"). In some
embodiments, an alkyl group has 1 to 7 carbon atoms ("C.sub.1-7
alkyl"). In some embodiments, an alkyl group has 1 to 6 carbon
atoms ("C.sub.1-6 alkyl"). In some embodiments, an alkyl group has
1 to 5 carbon atoms ("C.sub.1-5 alkyl"). In some embodiments, an
alkyl group has 1 to 4 carbon atoms ("C.sub.1-4 alkyl"). In some
embodiments, an alkyl group has 1 to 3 carbon atoms ("C.sub.1-3
alkyl"). In some embodiments, an alkyl group has 1 to 2 carbon
atoms ("C.sub.1-2 alkyl"). In some embodiments, an alkyl group has
1 carbon atom ("C.sub.1 alkyl"). In some embodiments, an alkyl
group has 2 to 6 carbon atoms ("C.sub.2-6 alkyl"). Examples of
C.sub.1-6 alkyl groups include methyl (C.sub.1), ethyl (C.sub.2),
propyl (C.sub.3) (e.g., n-propyl, isopropyl), butyl (C.sub.4)
(e.g., n-butyl, tert-butyl, sec-butyl, iso-butyl), pentyl (C.sub.5)
(e.g., n-pentyl, 3-pentanyl, amyl, neopentyl, 3-methyl-2-butanyl,
tertiary amyl), and hexyl (C.sub.6) (e.g., n-hexyl). Additional
examples of alkyl groups include n-heptyl (C.sub.7), n-octyl
(C.sub.8), and the like. Unless otherwise specified, each instance
of an alkyl group is independently unsubstituted (an "unsubstituted
alkyl") or substituted (a "substituted alkyl") with one or more
substituents (e.g., halogen, such as F). In certain embodiments,
the alkyl group is an unsubstituted C.sub.1-10 alkyl (such as
unsubstituted C.sub.1-6 alkyl, e.g., --CH.sub.3 (Me), unsubstituted
ethyl (Et), unsubstituted propyl (Pr, e.g., unsubstituted n-propyl
(n-Pr), unsubstituted isopropyl (i-Pr)), unsubstituted butyl (Bu,
e.g., unsubstituted n-butyl (n-Bu), unsubstituted tert-butyl
(tert-Bu or t-Bu), unsubstituted sec-butyl (sec-Bu), unsubstituted
isobutyl (i-Bu)). In certain embodiments, the alkyl group is a
substituted C.sub.1-10 alkyl (such as substituted C.sub.1-6 alkyl,
e.g., --CF.sub.3, Bn).
[0041] "Alkenyl" refers to a radical of a straight-chain or
branched hydrocarbon group having from 2 to 20 carbon atoms, one or
more carbon-carbon double bonds, and no triple bonds ("C.sub.2-20
alkenyl"). In some embodiments, an alkenyl group has 2 to 10 carbon
atoms ("C.sub.2-10 alkenyl"). In some embodiments, an alkenyl group
has 2 to 9 carbon atoms ("C.sub.2-9 alkenyl"). In some embodiments,
an alkenyl group has 2 to 8 carbon atoms ("C.sub.2-8 alkenyl"). In
some embodiments, an alkenyl group has 2 to 7 carbon atoms
("C.sub.2-7 alkenyl"). In some embodiments, an alkenyl group has 2
to 6 carbon atoms ("C.sub.2-6 alkenyl"). In some embodiments, an
alkenyl group has 2 to 5 carbon atoms ("C.sub.2-5 alkenyl"). In
some embodiments, an alkenyl group has 2 to 4 carbon atoms
("C.sub.2-4 alkenyl"). In some embodiments, an alkenyl group has 2
to 3 carbon atoms ("C.sub.2-3 alkenyl"). In some embodiments, an
alkenyl group has 2 carbon atoms ("C.sub.2 alkenyl"). The one or
more carbon-carbon double bonds can be internal (such as in
2-butenyl) or terminal (such as in 1-butenyl). Examples of
C.sub.2-4 alkenyl groups include ethenyl (C.sub.2), 1-propenyl
(C.sub.3), 2-propenyl (C.sub.3), 1-butenyl (C.sub.4), 2-butenyl
(C.sub.4), butadienyl (C.sub.4), and the like. Examples of
C.sub.2-6 alkenyl groups include the aforementioned C.sub.2-4
alkenyl groups as well as pentenyl (C.sub.5), pentadienyl
(C.sub.5), hexenyl (C.sub.6), and the like. Additional examples of
alkenyl include heptenyl (C.sub.7), octenyl (C.sub.8), octatrienyl
(C.sub.8), and the like. Unless otherwise specified, each instance
of an alkenyl group is independently optionally substituted, i.e.,
unsubstituted (an "unsubstituted alkenyl") or substituted (a
"substituted alkenyl") with one or more substituents. In certain
embodiments, the alkenyl group is unsubstituted C.sub.2-10 alkenyl.
In certain embodiments, the alkenyl group is substituted C.sub.2-10
alkenyl. In an alkenyl group, a C.dbd.C double bond for which the
stereochemistry is not specified (e.g., --CH.dbd.CHCH.sub.3 or
##STR00005##
may be an (E)- or (Z)-double bond.
[0042] "Alkynyl" refers to a radical of a straight-chain or
branched hydrocarbon group having from 2 to 20 carbon atoms, one or
more carbon-carbon triple bonds, and optionally one or more double
bonds ("C.sub.2-20 alkynyl"). In some embodiments, an alkynyl group
has 2 to 10 carbon atoms ("C.sub.2-10 alkynyl"). In some
embodiments, an alkynyl group has 2 to 9 carbon atoms ("C.sub.2-9
alkynyl"). In some embodiments, an alkynyl group has 2 to 8 carbon
atoms ("C.sub.2-8 alkynyl"). In some embodiments, an alkynyl group
has 2 to 7 carbon atoms ("C.sub.2-7 alkynyl"). In some embodiments,
an alkynyl group has 2 to 6 carbon atoms ("C.sub.2-6 alkynyl"). In
some embodiments, an alkynyl group has 2 to 5 carbon atoms
("C.sub.2-5 alkynyl"). In some embodiments, an alkynyl group has 2
to 4 carbon atoms ("C.sub.2-4 alkynyl"). In some embodiments, an
alkynyl group has 2 to 3 carbon atoms ("C.sub.2-3 alkynyl"). In
some embodiments, an alkynyl group has 2 carbon atoms ("C.sub.2
alkynyl"). The one or more carbon-carbon triple bonds can be
internal (such as in 2-butynyl) or terminal (such as in 1-butynyl).
Examples of C.sub.2-4 alkynyl groups include, without limitation,
ethynyl (C.sub.2), 1-propynyl (C.sub.3), 2-propynyl (C.sub.3),
1-butynyl (C.sub.4), 2-butynyl (C.sub.4), and the like. Examples of
C.sub.2-6 alkenyl groups include the aforementioned C.sub.2-4
alkynyl groups as well as pentynyl (C.sub.5), hexynyl (C.sub.6),
and the like. Additional examples of alkynyl include heptynyl
(C.sub.7), octynyl (C.sub.8), and the like. Unless otherwise
specified, each instance of an alkynyl group is independently
optionally substituted, i.e., unsubstituted (an "unsubstituted
alkynyl") or substituted (a "substituted alkynyl") with one or more
substituents. In certain embodiments, the alkynyl group is
unsubstituted C.sub.2-10 alkynyl. In certain embodiments, the
alkynyl group is substituted C.sub.2-10 alkynyl.
[0043] "Carbocyclyl" or "carbocyclic" refers to a radical of a
non-aromatic cyclic hydrocarbon group having from 3 to 10 ring
carbon atoms ("C.sub.3-10 carbocyclyl") and zero heteroatoms in the
non-aromatic ring system. In some embodiments, a carbocyclyl group
has 3 to 8 ring carbon atoms ("C.sub.3-8 carbocyclyl"). In some
embodiments, a carbocyclyl group has 3 to 6 ring carbon atoms
("C.sub.3-6 carbocyclyl"). In some embodiments, a carbocyclyl group
has 3 to 6 ring carbon atoms ("C.sub.3-6 carbocyclyl"). In some
embodiments, a carbocyclyl group has 5 to 10 ring carbon atoms
("C.sub.5-10 carbocyclyl"). Exemplary C.sub.3-6 carbocyclyl groups
include, without limitation, cyclopropyl (C.sub.3), cyclopropenyl
(C.sub.3), cyclobutyl (C.sub.4), cyclobutenyl (C.sub.4),
cyclopentyl (C.sub.5), cyclopentenyl (C.sub.5), cyclohexyl
(C.sub.6), cyclohexenyl (C.sub.6), cyclohexadienyl (C.sub.6), and
the like. Exemplary C.sub.3-8 carbocyclyl groups include, without
limitation, the aforementioned C.sub.3-6 carbocyclyl groups as well
as cycloheptyl (C.sub.7), cycloheptenyl (C.sub.7), cycloheptadienyl
(C.sub.7), cycloheptatrienyl (C.sub.7), cyclooctyl (C.sub.8),
cyclooctenyl (C.sub.8), bicyclo[2.2.1]heptanyl (C.sub.7),
bicyclo[2.2.2]octanyl (C.sub.8), and the like. Exemplary C.sub.3-10
carbocyclyl groups include, without limitation, the aforementioned
C.sub.3-8 carbocyclyl groups as well as cyclononyl (C.sub.9),
cyclononenyl (C.sub.9), cyclodecyl (C.sub.10), cyclodecenyl
(C.sub.10), octahydro-1H-indenyl (C.sub.9), decahydronaphthalenyl
(C.sub.10), spiro[4.5]decanyl (C.sub.10), and the like. As the
foregoing examples illustrate, in certain embodiments, the
carbocyclyl group is either monocyclic ("monocyclic carbocyclyl")
or contain a fused, bridged or spiro ring system such as a bicyclic
system ("bicyclic carbocyclyl") and can be saturated or can be
partially unsaturated. "Carbocyclyl" also includes ring systems
wherein the carbocyclic ring, as defined above, is fused with one
or more aryl or heteroaryl groups wherein the point of attachment
is on the carbocyclic ring, and in such instances, the number of
carbons continue to designate the number of carbons in the
carbocyclic ring system. Unless otherwise specified, each instance
of a carbocyclyl group is independently optionally substituted,
i.e., unsubstituted (an "unsubstituted carbocyclyl") or substituted
(a "substituted carbocyclyl") with one or more substituents. In
certain embodiments, the carbocyclyl group is unsubstituted
C.sub.3-10 carbocyclyl. In certain embodiments, the carbocyclyl
group is substituted C.sub.3-10 carbocyclyl.
[0044] In some embodiments, "carbocyclyl" is a monocyclic,
saturated carbocyclyl group having from 3 to 10 ring carbon atoms
("C.sub.3-10 cycloalkyl"). In some embodiments, a cycloalkyl group
has 3 to 8 ring carbon atoms ("C.sub.3-8 cycloalkyl"). In some
embodiments, a cycloalkyl group has 3 to 6 ring carbon atoms
("C.sub.3-6 cycloalkyl"). In some embodiments, a cycloalkyl group
has 5 to 6 ring carbon atoms ("C.sub.5-6 cycloalkyl"). In some
embodiments, a cycloalkyl group has 5 to 10 ring carbon atoms
("C.sub.5-10 cycloalkyl"). Examples of C.sub.5-6 cycloalkyl groups
include cyclopentyl (C.sub.5) and cyclohexyl (C.sub.5). Examples of
C.sub.3-6 cycloalkyl groups include the aforementioned C.sub.5-6
cycloalkyl groups as well as cyclopropyl (C.sub.3) and cyclobutyl
(C.sub.4). Examples of C.sub.3-8 cycloalkyl groups include the
aforementioned C.sub.3-6 cycloalkyl groups as well as cycloheptyl
(C.sub.7) and cyclooctyl (C.sub.8). Unless otherwise specified,
each instance of a cycloalkyl group is independently unsubstituted
(an "unsubstituted cycloalkyl") or substituted (a "substituted
cycloalkyl") with one or more substituents. In certain embodiments,
the cycloalkyl group is unsubstituted C.sub.3-10 cycloalkyl. In
certain embodiments, the cycloalkyl group is substituted C.sub.3-10
cycloalkyl.
[0045] "Heterocyclyl" or "heterocyclic" refers to a radical of a 3-
to 10-membered non-aromatic ring system having ring carbon atoms
and 1 to 4 ring heteroatoms, wherein each heteroatom is
independently selected from nitrogen, oxygen, sulfur, boron,
phosphorus, and silicon ("3-10 membered heterocyclyl"). In
heterocyclyl groups that contain one or more nitrogen atoms, the
point of attachment can be a carbon or nitrogen atom, as valency
permits. A heterocyclyl group can either be monocyclic ("monocyclic
heterocyclyl") or a fused, bridged, or spiro ring system, such as a
bicyclic system ("bicyclic heterocyclyl"), and can be saturated or
can be partially unsaturated. Heterocyclyl bicyclic ring systems
can include one or more heteroatoms in one or both rings.
"Heterocyclyl" also includes ring systems wherein the heterocyclic
ring, as defined above, is fused with one or more carbocyclyl
groups wherein the point of attachment is either on the carbocyclyl
or heterocyclic ring, or ring systems wherein the heterocyclic
ring, as defined above, is fused with one or more aryl or
heteroaryl groups, wherein the point of attachment is on the
heterocyclic ring, and in such instances, the number of ring
members continue to designate the number of ring members in the
heterocyclic ring system. Unless otherwise specified, each instance
of heterocyclyl is independently optionally substituted, i.e.,
unsubstituted (an "unsubstituted heterocyclyl") or substituted (a
"substituted heterocyclyl") with one or more substituents. In
certain embodiments, the heterocyclyl group is unsubstituted 3-10
membered heterocyclyl. In certain embodiments, the heterocyclyl
group is substituted 3-10 membered heterocyclyl.
[0046] In some embodiments, a heterocyclyl group is a 5-10
membered, non-aromatic ring system having ring carbon atoms and 1-4
ring heteroatoms, wherein each heteroatom is independently selected
from nitrogen, oxygen, sulfur, boron, phosphorus, and silicon
("5-10 membered heterocyclyl"). In some embodiments, a heterocyclyl
group is a 5-8 membered non-aromatic ring system having ring carbon
atoms and 1-4 ring heteroatoms, wherein each heteroatom is
independently selected from nitrogen, oxygen, and sulfur ("5-8
membered heterocyclyl"). In some embodiments, a heterocyclyl group
is a 5-6 membered non-aromatic ring system having ring carbon atoms
and 1-4 ring heteroatoms, wherein each heteroatom is independently
selected from nitrogen, oxygen, and sulfur ("5-6 membered
heterocyclyl"). In some embodiments, the 5-6 membered heterocyclyl
has 1-3 ring heteroatoms selected from nitrogen, oxygen, and
sulfur. In some embodiments, the 5-6 membered heterocyclyl has 1-2
ring heteroatoms selected from nitrogen, oxygen, and sulfur. In
some embodiments, the 5-6 membered heterocyclyl has one ring
heteroatom selected from nitrogen, oxygen, and sulfur.
[0047] Exemplary 3-membered heterocyclyl groups containing one
heteroatom include, without limitation, azirdinyl, oxiranyl,
thiiranyl. Exemplary 4-membered heterocyclyl groups containing one
heteroatom include, without limitation, azetidinyl, oxetanyl and
thietanyl. Exemplary 5-membered heterocyclyl groups containing one
heteroatom include, without limitation, tetrahydrofuranyl,
dihydrofuranyl, tetrahydrothiophenyl, dihydrothiophenyl,
pyrrolidinyl, dihydropyrrolyl, and pyrrolyl-2,5-dione. Exemplary
5-membered heterocyclyl groups containing two heteroatoms include,
without limitation, dioxolanyl, oxasulfuranyl, disulfuranyl, and
oxazolidin-2-one. Exemplary 5-membered heterocyclyl groups
containing three heteroatoms include, without limitation,
triazolinyl, oxadiazolinyl, and thiadiazolinyl. Exemplary
6-membered heterocyclyl groups containing one heteroatom include,
without limitation, piperidinyl, tetrahydropyranyl,
dihydropyridinyl, and thianyl. Exemplary 6-membered heterocyclyl
groups containing two heteroatoms include, without limitation,
piperazinyl, morpholinyl, dithianyl, and dioxanyl. Exemplary
6-membered heterocyclyl groups containing two heteroatoms include,
without limitation, triazinanyl. Exemplary 7-membered heterocyclyl
groups containing one heteroatom include, without limitation,
azepanyl, oxepanyl and thiepanyl. Exemplary 8-membered heterocyclyl
groups containing one heteroatom include, without limitation,
azocanyl, oxecanyl and thiocanyl. Exemplary 5-membered heterocyclyl
groups fused to a C.sub.6 aryl ring (also referred to herein as a
5,6-bicyclic heterocyclic ring) include, without limitation,
indolinyl, isoindolinyl, dihydrobenzofuranyl, dihydrobenzothienyl,
benzoxazolinonyl, and the like. Exemplary 6-membered heterocyclyl
groups fused to an aryl ring (also referred to herein as a
6,6-bicyclic heterocyclic ring) include, without limitation,
tetrahydroquinolinyl, tetrahydroisoquinolinyl, and the like.
[0048] "Aryl" refers to a radical of a monocyclic or polycyclic
(e.g., bicyclic or tricyclic) 4n+2 aromatic ring system (e.g.,
having 6, 10, or 14 pi electrons shared in a cyclic array) having
6-14 ring carbon atoms and zero heteroatoms provided in the
aromatic ring system ("C.sub.6-14 aryl"). In some embodiments, an
aryl group has six ring carbon atoms ("C.sub.6 aryl"; e.g.,
phenyl). In some embodiments, an aryl group has ten ring carbon
atoms ("C.sub.10 aryl"; e.g., naphthyl such as 1-naphthyl and
2-naphthyl). In some embodiments, an aryl group has fourteen ring
carbon atoms ("C.sub.14 aryl"; e.g., anthracyl). "Aryl" also
includes ring systems wherein the aryl ring, as defined above, is
fused with one or more carbocyclyl or heterocyclyl groups, wherein
the radical or point of attachment is on the aryl ring, and in such
instances, the number of carbon atoms continue to designate the
number of carbon atoms in the aryl ring system. Unless otherwise
specified, each instance of an aryl group is independently
optionally substituted, i.e., unsubstituted (an "unsubstituted
aryl") or substituted (a "substituted aryl") with one or more
substituents. In certain embodiments, the aryl group is
unsubstituted C.sub.6-14 aryl. In certain embodiments, the aryl
group is substituted C.sub.6-14 aryl.
[0049] "Aralkyl" refers to an optionally substituted alkyl group
substituted by an optionally substituted aryl group. In certain
embodiments, the aralkyl is optionally substituted benzyl. In
certain embodiments, the aralkyl is benzyl. In certain embodiments,
the aralkyl is optionally substituted phenethyl. In certain
embodiments, the aralkyl is phenethyl.
[0050] "Heteroaryl" refers to a radical of a 5-10 membered,
monocyclic or bicyclic 4n+2 aromatic ring system (e.g., having 6 or
10 pi electrons shared in a cyclic array) having ring carbon atoms
and 1-4 ring heteroatoms provided in the aromatic ring system,
wherein each heteroatom is independently selected from nitrogen,
oxygen and sulfur ("5-10 membered heteroaryl"). In heteroaryl
groups that contain one or more nitrogen atoms, the point of
attachment can be a carbon or nitrogen atom, as valency permits.
Heteroaryl bicyclic ring systems can include one or more
heteroatoms in one or both rings. "Heteroaryl" includes ring
systems wherein the heteroaryl ring, as defined above, is fused
with one or more carbocyclyl or heterocyclyl groups wherein the
point of attachment is on the heteroaryl ring, and in such
instances, the number of ring members continue to designate the
number of ring members in the heteroaryl ring system. "Heteroaryl"
also includes ring systems wherein the heteroaryl ring, as defined
above, is fused with one or more aryl groups wherein the point of
attachment is either on the aryl or heteroaryl ring, and in such
instances, the number of ring members designates the number of ring
members in the fused (aryl/heteroaryl) ring system. Bicyclic
heteroaryl groups wherein one ring does not contain a heteroatom
(e.g., indolyl, quinolinyl, carbazolyl, and the like) the point of
attachment can be on either ring, i.e., either the ring bearing a
heteroatom (e.g., 2-indolyl) or the ring that does not contain a
heteroatom (e.g., 5-indolyl).
[0051] In some embodiments, a heteroaryl group is a 5-10 membered
aromatic ring system having ring carbon atoms and 1-4 ring
heteroatoms provided in the aromatic ring system, wherein each
heteroatom is independently selected from nitrogen, oxygen, and
sulfur ("5-10 membered heteroaryl"). In some embodiments, a
heteroaryl group is a 5-8 membered aromatic ring system having ring
carbon atoms and 1-4 ring heteroatoms provided in the aromatic ring
system, wherein each heteroatom is independently selected from
nitrogen, oxygen, and sulfur ("5-8 membered heteroaryl"). In some
embodiments, a heteroaryl group is a 5-6 membered aromatic ring
system having ring carbon atoms and 1-4 ring heteroatoms provided
in the aromatic ring system, wherein each heteroatom is
independently selected from nitrogen, oxygen, and sulfur ("5-6
membered heteroaryl"). In some embodiments, the 5-6 membered
heteroaryl has 1-3 ring heteroatoms selected from nitrogen, oxygen,
and sulfur. In some embodiments, the 5-6 membered heteroaryl has
1-2 ring heteroatoms selected from nitrogen, oxygen, and sulfur. In
some embodiments, the 5-6 membered heteroaryl has 1 ring heteroatom
selected from nitrogen, oxygen, and sulfur. Unless otherwise
specified, each instance of a heteroaryl group is independently
optionally substituted, i.e., unsubstituted (an "unsubstituted
heteroaryl") or substituted (a "substituted heteroaryl") with one
or more substituents. In certain embodiments, the heteroaryl group
is unsubstituted 5-14 membered heteroaryl. In certain embodiments,
the heteroaryl group is substituted 5-14 membered heteroaryl.
[0052] Exemplary 5-membered heteroaryl groups containing one
heteroatom include, without limitation, pyrrolyl, furanyl, and
thiophenyl. Exemplary 5-membered heteroaryl groups containing two
heteroatoms include, without limitation, imidazolyl, pyrazolyl,
oxazolyl, isoxazolyl, thiazolyl, and isothiazolyl. Exemplary
5-membered heteroaryl groups containing three heteroatoms include,
without limitation, triazolyl, oxadiazolyl, and thiadiazolyl.
Exemplary 5-membered heteroaryl groups containing four heteroatoms
include, without limitation, tetrazolyl. Exemplary 6-membered
heteroaryl groups containing one heteroatom include, without
limitation, pyridinyl. Exemplary 6-membered heteroaryl groups
containing two heteroatoms include, without limitation,
pyridazinyl, pyrimidinyl, and pyrazinyl. Exemplary 6-membered
heteroaryl groups containing three or four heteroatoms include,
without limitation, triazinyl and tetrazinyl, respectively.
Exemplary 7-membered heteroaryl groups containing one heteroatom
include, without limitation, azepinyl, oxepinyl, and thiepinyl.
Exemplary 5,6-bicyclic heteroaryl groups include, without
limitation, indolyl, isoindolyl, indazolyl, benzotriazolyl,
benzothiophenyl, isobenzothiophenyl, benzofuranyl, benzoisofuranyl,
benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzoxadiazolyl,
benzthiazolyl, benzisothiazolyl, benzthiadiazolyl, indolizinyl, and
purinyl. Exemplary 6,6-bicyclic heteroaryl groups include, without
limitation, naphthyridinyl, pteridinyl, quinolinyl, isoquinolinyl,
cinnolinyl, quinoxalinyl, phthalazinyl, and quinazolinyl.
[0053] "Heteroaralkyl" is a subset of alkyl and heteroaryl and
refers to an optionally substituted alkyl group substituted by an
optionally substituted heteroaryl group.
[0054] "Unsaturated" or "partially unsaturated" refers to a group
that includes at least one double or triple bond. A "partially
unsaturated" ring system is further intended to encompass rings
having multiple sites of unsaturation, but is not intended to
include aromatic groups (e.g., aryl or heteroaryl groups).
Likewise, "saturated" refers to a group that does not contain a
double or triple bond, i.e., contains all single bonds.
[0055] Alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl,
and heteroaryl groups, which are divalent linking groups, are
further referred to using the suffix-ene, e.g., alkylene,
alkenylene, alkynylene, carbocyclylene, heterocyclylene, arylene,
and heteroarylene.
[0056] An atom, moiety, or group described herein may be
unsubstituted or substituted, as valency permits, unless otherwise
provided expressly. The term "optionally substituted" refers to
substituted or unsubstituted.
[0057] A group is optionally substituted unless expressly provided
otherwise. The term "optionally substituted" refers to being
substituted or unsubstituted. In certain embodiments, alkyl,
alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl
groups are optionally substituted (e.g., "substituted" or
"unsubstituted" alkyl, "substituted" or "unsubstituted" alkenyl,
"substituted" or "unsubstituted" alkynyl, "substituted" or
"unsubstituted" carbocyclyl, "substituted" or "unsubstituted"
heterocyclyl, "substituted" or "unsubstituted" aryl or
"substituted" or "unsubstituted" heteroaryl group). In general, the
term "substituted", whether preceded by the term "optionally" or
not, means that at least one hydrogen present on a group (e.g., a
carbon or nitrogen atom) is replaced with a permissible
substituent, e.g., a substituent which upon substitution results in
a stable compound, e.g., a compound which does not spontaneously
undergo transformation such as by rearrangement, cyclization,
elimination, or other reaction. Unless otherwise indicated, a
"substituted" group has a substituent at one or more substitutable
positions of the group, and when more than one position in any
given structure is substituted, the substituent is either the same
or different at each position. The term "substituted" is
contemplated to include substitution with all permissible
substituents of organic compounds, any of the substituents
described herein that results in the formation of a stable
compound. The present disclosure contemplates any and all such
combinations in order to arrive at a stable compound. For purposes
of this disclosure, heteroatoms such as nitrogen may have hydrogen
substituents and/or any suitable substituent as described herein
which satisfy the valencies of the heteroatoms and results in the
formation of a stable moiety. In certain embodiments, the
substituent is a carbon atom substituent. In certain embodiments,
the substituent is a nitrogen atom substituent. In certain
embodiments, the substituent is an oxygen atom substituent. In
certain embodiments, the substituent is a sulfur atom
substituent.
[0058] Exemplary carbon atom substituents include, but are not
limited to, halogen, --CN, --NO.sub.2, --N.sub.3, --SO.sub.2H,
--SO.sub.3H, --OH, --OR.sup.aa, --ON(R.sup.bb).sub.2,
--N(R.sup.bb).sub.2, --N(R.sup.bb).sub.3.sup.+X.sup.-,
--N(OR.sup.cc)R.sup.bb, --SH, --SR.sup.aa, --SSR.sup.cc,
--C(.dbd.O)R.sup.aa, --CO.sub.2H, --CHO, --C(OR.sup.cc).sub.2,
--CO.sub.2R.sup.aa, --OC(.dbd.O)R.sup.aa, --OCO.sub.2R.sup.aa,
--C(.dbd.O)N(R.sup.bb).sub.2, --OC(.dbd.O)N(R.sup.bb).sub.2,
--NR.sup.bbC(.dbd.O)R.sup.aa, --NR.sup.bbCO.sub.2R.sup.aa,
--NR.sup.bbC(.dbd.O)N(R.sup.bb).sub.2, --C(.dbd.NR.sup.bb)R.sup.aa,
--C(.dbd.NR.sup.bb)OR.sup.aa, --OC(.dbd.NR.sup.bb)R.sup.aa,
--OC(.dbd.NR.sup.bb)OR.sup.aa,
--C(.dbd.NR.sup.bb)N(R.sup.bb).sub.2,
--OC(.dbd.NR.sup.bb)N(R.sup.bb).sub.2,
--NR.sup.bbC(.dbd.NR.sup.bb)N(R.sup.bb).sub.2,
--C(.dbd.O)NR.sup.bbSO.sub.2R.sup.aa, --NR.sup.bbSO.sub.2R.sup.aa,
--SO.sub.2N(R.sup.bb).sub.2, --SO.sub.2R.sup.aa,
--SO.sub.2OR.sup.aa, --OSO.sub.2R.sup.aa, --S(.dbd.O)R.sup.aa,
--OS(.dbd.O)R.sup.aa, --Si(R.sup.aa).sub.3,
--OSi(R.sup.aa).sub.3--C(.dbd.S)N(R.sup.bb).sub.2,
--C(.dbd.O)SR.sup.aa, --C(.dbd.S)SR.sup.aa, --SC(.dbd.S)SR.sup.aa,
--SC(.dbd.O)SR.sup.aa, --OC(.dbd.O)SR.sup.aa,
--SC(.dbd.O)OR.sup.aa, --SC(.dbd.O)R.sup.aa,
--P(.dbd.O)(R.sup.aa).sub.2, --P(.dbd.O)(OR.sup.cc).sub.2,
--OP(.dbd.O)(R.sup.aa).sub.2, --OP(.dbd.O)(OR.sup.cc).sub.2,
--P(.dbd.O)(N(R.sup.bb).sub.2).sub.2,
--OP(.dbd.O)(N(R.sup.bb).sub.2).sub.2,
--NR.sup.bbP(.dbd.O)(R.sup.aa).sub.2,
--NR.sup.bbP(.dbd.O)(OR.sup.cc).sub.2,
--NR.sup.bbP(.dbd.O)(N(R.sup.bb).sub.2).sub.2, --P(R.sup.cc).sub.2,
--P(OR.sup.cc).sub.2, --P(R.sup.cc).sub.3.sup.+X.sup.-,
--P(OR.sup.cc).sub.3.sup.+X.sup.-, --P(R.sup.cc).sub.4,
--P(OR.sup.cc).sub.4, --OP(R.sup.cc).sub.2,
--OP(R.sup.cc).sub.3.sup.+X.sup.-, --OP(OR.sup.cc).sub.2,
--OP(OR.sup.cc).sub.3.sup.+X.sup.-, --OP(R.sup.cc).sub.4,
--OP(OR.sup.cc).sub.4, --B(R.sup.aa).sub.2, --B(OR.sup.cc).sub.2,
--BR.sup.aa(OR.sup.cc), C.sub.1-10 alkyl, C.sub.1-10 perhaloalkyl,
C.sub.2-10 alkenyl, C.sub.2-10 alkynyl, heteroC.sub.1-10 alkyl,
heteroC.sub.2-10 alkenyl, heteroC.sub.2-10 alkynyl, C.sub.3-10
carbocyclyl, 3-14 membered heterocyclyl, C.sub.6-14 aryl, and 5-14
membered heteroaryl, wherein each alkyl, alkenyl, alkynyl,
heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl,
heterocyclyl, aryl, and heteroaryl is independently substituted
with 0, 1, 2, 3, 4, or 5 R.sup.dd groups; wherein X.sup.- is a
counterion;
[0059] or two geminal hydrogens on a carbon atom are replaced with
the group .dbd.O, .dbd.S, .dbd.NN(R.sup.bb).sub.2,
.dbd.NNR.sup.bbC(.dbd.O)R.sup.aa,
.dbd.NNR.sup.bbC(.dbd.O)OR.sup.aa,
.dbd.NNR.sup.bbS(.dbd.O).sub.2R.sup.aa, .dbd.NR.sup.bb, or
.dbd.NOR.sup.cc;
[0060] each instance of R.sup.aa is, independently, selected from
C.sub.1-10 alkyl, C.sub.1-10 perhaloalkyl, C.sub.2-10 alkenyl,
C.sub.2-10 alkynyl, C.sub.3-10 carbocyclyl, 3-14 membered
heterocyclyl, C.sub.6-14 aryl, and 5-14 membered heteroaryl, or two
R.sup.aa groups are joined to form a 3-14 membered heterocyclyl or
5-14 membered heteroaryl ring, wherein each alkyl, alkenyl,
alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is
independently substituted with 0, 1, 2, 3, 4, or 5 R.sup.dd
groups;
[0061] each instance of R.sup.bb is, independently, selected from
hydrogen, --OH, --OR.sup.aa, --N(R.sup.cc).sub.2, --CN,
--C(.dbd.O)R.sup.aa, --C(.dbd.O)N(R.sup.cc).sub.2,
--CO.sub.2R.sup.aa, --SO.sub.2R.sup.aa,
--C(.dbd.NR.sup.cc)OR.sup.aa, --C(.dbd.NR.sup.cc)N(R.sup.cc).sub.2,
--SO.sub.2N(R.sup.cc).sub.2, --SO.sub.2R.sup.cc,
--SO.sub.2OR.sup.cc, --SOR.sup.aa, --C(.dbd.S)N(R.sup.cc).sub.2,
--C(.dbd.O)SR.sup.cc, --C(.dbd.S)SR.sup.cc,
--P(.dbd.O)(R.sup.aa).sub.2, --P(.dbd.O)(OR.sup.cc).sub.2,
--P(.dbd.O)(N(R.sup.cc).sub.2).sub.2, C.sub.1-10 alkyl, C.sub.1-10
perhaloalkyl, C.sub.2-10 alkenyl, C.sub.2-10 alkynyl,
heteroC.sub.1-10alkyl, heteroC.sub.2-10alkenyl,
heteroC.sub.2-10alkynyl, C.sub.3-10 carbocyclyl, 3-14 membered
heterocyclyl, C.sub.6-14 aryl, and 5-14 membered heteroaryl, or two
R.sup.bb groups are joined to form a 3-14 membered heterocyclyl or
5-14 membered heteroaryl ring, wherein each alkyl, alkenyl,
alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl,
heterocyclyl, aryl, and heteroaryl is independently substituted
with 0, 1, 2, 3, 4, or 5 R.sup.dd groups; wherein X.sup.- is a
counterion;
[0062] each instance of R.sup.cc is, independently, selected from
hydrogen, C.sub.1-10 alkyl, C.sub.1-10 perhaloalkyl, C.sub.2-10
alkenyl, C.sub.2-10 alkynyl, C.sub.3-10 carbocyclyl, 3-14 membered
heterocyclyl, C.sub.6-14 aryl, and 5-14 membered heteroaryl, or two
R.sup.cc groups are joined to form a 3-14 membered heterocyclyl or
5-14 membered heteroaryl ring, wherein each alkyl, alkenyl,
alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is
independently substituted with 0, 1, 2, 3, 4, or 5 R.sup.dd
groups;
[0063] each instance of R.sup.dd is, independently, selected from
halogen, --CN, --NO.sub.2, --N.sub.3, --SO.sub.2H, --SO.sub.3H,
--OH, --OR.sup.ee, --ON(R.sup.ff).sub.2, --N(R.sup.ff).sub.2,
--N(R.sup.ff).sub.3.sup.+X.sup.-, --N(OR.sup.ee)R.sup.ff, --SH,
--SR.sup.ee, --SSR.sup.ee, --C(.dbd.O)R.sup.ee, --CO.sub.2H,
--CO.sub.2R.sup.ee, --OC(.dbd.O)R.sup.ee, --OCO.sub.2R.sup.ee,
--C(.dbd.O)N(R.sup.ff).sub.2, --OC(.dbd.O)N(R.sup.ff).sub.2,
--NR.sup.ffC(.dbd.O)R.sup.ee, --NR.sup.ffCO.sub.2R.sup.ee,
--NR.sup.ffC(.dbd.O)N(R.sup.ff).sub.2,
--C(.dbd.NR.sup.ff)OR.sup.ee, --OC(.dbd.NR.sup.ff)R.sup.ee,
--OC(.dbd.NR.sup.ff)OR.sup.ee,
--C(.dbd.NR.sup.ff)N(R.sup.ff).sub.2,
--OC(.dbd.NR.sup.ff)N(R.sup.ff).sub.2,
--NR.sup.ffC(.dbd.NR.sup.ff)N(R.sup.ff).sub.2,
--NR.sup.ffSO.sub.2R.sup.ee, --SO.sub.2N(R.sup.ff).sub.2,
--SO.sub.2R.sup.ee, --SO.sub.2OR.sup.ee, --OSO.sub.2R.sup.ee,
--S(.dbd.O)R.sup.ee, --Si(R.sup.ee).sub.3, --OSi(R.sup.ee).sub.3,
--C(.dbd.S)N(R.sup.ff).sub.2, --C(.dbd.O)SR.sup.ee,
--C(.dbd.S)SR.sup.ee, --SC(.dbd.S)SR.sup.ee,
--P(.dbd.O)(OR.sup.ee).sub.2, --P(.dbd.O)(R.sup.ee).sub.2,
--OP(.dbd.O)(R.sup.ee).sub.2, --OP(.dbd.O)(OR.sup.ee).sub.2,
C.sub.1-6 alkyl, C.sub.1-6 perhaloalkyl, C.sub.2-6 alkenyl,
C.sub.2-6 alkynyl, heteroC.sub.1-6alkyl, heteroC.sub.2-6alkenyl,
heteroC.sub.2-6alkynyl, C.sub.3-10 carbocyclyl, 3-10 membered
heterocyclyl, C.sub.6-10 aryl, 5-10 membered heteroaryl, wherein
each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl,
heteroalkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is
independently substituted with 0, 1, 2, 3, 4, or 5 R.sup.gg groups,
or two geminal R.sup.dd substituents can be joined to form .dbd.O
or .dbd.S; wherein X.sup.- is a counterion;
[0064] each instance of R.sup.ee is, independently, selected from
C.sub.1-6 alkyl, C.sub.1-6 perhaloalkyl, C.sub.2-6 alkenyl,
C.sub.2-6 alkynyl, C.sub.3-10 carbocyclyl, C.sub.6-10 aryl, 3-10
membered heterocyclyl, and 3-10 membered heteroaryl, wherein each
alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and
heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5
R.sup.gg groups;
[0065] each instance of R.sup.ff is, independently, selected from
hydrogen, C.sub.1-6 alkyl, C.sub.1-6 perhaloalkyl, C.sub.2-6
alkenyl, C.sub.2-6 alkynyl, C.sub.3-10 carbocyclyl, 3-10 membered
heterocyclyl, C.sub.6-10 aryl and 5-10 membered heteroaryl, or two
R.sup.ff groups are joined to form a 3-14 membered heterocyclyl or
5-14 membered heteroaryl ring, wherein each alkyl, alkenyl,
alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is
independently substituted with 0, 1, 2, 3, 4, or 5 R.sup.gg groups;
and
[0066] each instance of R.sup.gg is, independently, halogen, --CN,
--NO.sub.2, --N.sub.3, --SO.sub.2H, --SO.sub.3H, --OH, --OC.sub.1-6
alkyl, --ON(C.sub.1-6 alkyl).sub.2, --N(C.sub.1-6 alkyl).sub.2,
--N(C.sub.1-6 alkyl).sub.3.sup.+X.sup.-, --NH(C.sub.1-6
alkyl).sub.2.sup.+X.sup.-, --NH.sub.2(C.sub.1-6
alkyl).sup.+X.sup.-, --NH.sub.3.sup.+X.sup.-, --N(OC.sub.1-6
alkyl)(C.sub.1-6 alkyl), --N(OH)(C.sub.1-6 alkyl), --NH(OH), --SH,
--SC.sub.1-6 alkyl, --SS(C.sub.1-6 alkyl), --C(.dbd.O)(C.sub.1-6
alkyl), --CO.sub.2H, --CO.sub.2(C.sub.1-6 alkyl),
--OC(.dbd.O)(C.sub.1-6 alkyl), --OCO.sub.2(C.sub.1-6 alkyl),
--C(.dbd.O)NH.sub.2, --C(.dbd.O)N(C.sub.1-6 alkyl).sub.2,
--OC(.dbd.O)NH(C.sub.1-6 alkyl), --NHC(.dbd.O)(C.sub.1-6 alkyl),
--N(C.sub.1-6 alkyl)C(.dbd.O)(C.sub.1-6 alkyl),
--NHCO.sub.2(C.sub.1-6 alkyl), --NHC(.dbd.O)N(C.sub.1-6
alkyl).sub.2, --NHC(.dbd.O)NH(C.sub.1-6 alkyl),
--NHC(.dbd.O)NH.sub.2, --C(.dbd.NH)O(C.sub.1-6 alkyl),
--OC(.dbd.NH)(C.sub.1-6 alkyl), --OC(.dbd.NH)OC.sub.1-6 alkyl,
--C(.dbd.NH)N(C.sub.1-6 alkyl).sub.2, --C(.dbd.NH)NH(C.sub.1-6
alkyl), --C(.dbd.NH)NH.sub.2, --OC(.dbd.NH)N(C.sub.1-6
alkyl).sub.2, --OC(NH)NH(C.sub.1-6 alkyl), --OC(NH)NH.sub.2,
--NHC(NH)N(C.sub.1-6 alkyl).sub.2, --NHC(.dbd.NH)NH.sub.2,
--NHSO.sub.2(C.sub.1-6 alkyl), --SO.sub.2N(C.sub.1-6 alkyl).sub.2,
--SO.sub.2NH(C.sub.1-6 alkyl), --SO.sub.2NH.sub.2,
--SO.sub.2C.sub.1-6 alkyl, --SO.sub.2OC.sub.1-6 alkyl,
--OSO.sub.2C.sub.1-6 alkyl, --SOC.sub.1-6 alkyl, --Si(C.sub.1-6
alkyl).sub.3, --OSi(C.sub.1-6 alkyl).sub.3-C(.dbd.S)N(C.sub.1-6
alkyl).sub.2, C(.dbd.S)NH(C.sub.1-6 alkyl), C(.dbd.S)NH.sub.2,
--C(.dbd.O)S(C.sub.1-6 alkyl), --C(.dbd.S)SC.sub.1-6 alkyl,
--SC(.dbd.S)SC.sub.1-6 alkyl, --P(.dbd.O)(OC.sub.1-6 alkyl).sub.2,
--P(.dbd.O)(C.sub.1-6 alkyl).sub.2, --OP(.dbd.O)(C.sub.1-6
alkyl).sub.2, --OP(.dbd.O)(OC.sub.1-6 alkyl).sub.2, C.sub.1-6
alkyl, C.sub.1-6 perhaloalkyl, C.sub.2-6 alkenyl, C.sub.2-6
alkynyl, heteroC.sub.1-6alkyl, heteroC.sub.2-6alkenyl,
heteroC.sub.2-6alkynyl, C.sub.3-10 carbocyclyl, C.sub.6-10 aryl,
3-10 membered heterocyclyl, 5-10 membered heteroaryl; or two
geminal R.sup.gg substituents can be joined to form .dbd.O or
.dbd.S; wherein X is a counterion.
[0067] A "counterion" or "anionic counterion" is a negatively
charged group associated with a positively charged group in order
to maintain electronic neutrality. An anionic counterion may be
monovalent (i.e., including one formal negative charge). An anionic
counterion may also be multivalent (i.e., including more than one
formal negative charge), such as divalent or trivalent. Exemplary
counterions include halide ions (e.g., F.sup.-, Cl.sup.-, Br.sup.-,
I.sup.-), NO.sub.3.sup.-, ClO.sub.4.sup.-, OH.sup.-,
H.sub.2PO.sub.4.sup.-, HCO.sub.3.sup.-, HSO.sub.4.sup.-, sulfonate
ions (e.g., methansulfonate, trifluoromethanesulfonate,
p-toluenesulfonate, benzenesulfonate, 10-camphor sulfonate,
naphthalene-2-sulfonate, naphthalene-1-sulfonic acid-5-sulfonate,
ethan-1-sulfonic acid-2-sulfonate, and the like), carboxylate ions
(e.g., acetate, propanoate, benzoate, glycerate, lactate, tartrate,
glycolate, gluconate, and the like), BF.sub.4.sup.-,
PF.sub.4.sup.-, PF.sub.6.sup.-, AsF.sub.6.sup.-, SbF.sub.6.sup.-,
B[3,5-(CF.sub.3).sub.2C.sub.6H.sub.3].sub.4].sup.-,
B(C.sub.6F.sub.5).sub.4.sup.-, BPh.sub.4.sup.-,
Al(OC(CF.sub.3).sub.3).sub.4.sup.-, and carborane anions (e.g.,
CB.sub.11H.sub.12.sup.- or (HCB.sub.11Me.sub.5Br.sub.6).sup.-).
Exemplary counterions which may be multivalent include
CO.sub.3.sup.2-, HPO.sub.4.sup.2-, PO.sub.4.sup.3-,
B.sub.4O.sub.7.sup.2-, SO.sub.4.sup.2-, S.sub.2O.sub.3.sup.2-,
carboxylate anions (e.g., tartrate, citrate, fumarate, maleate,
malate, malonate, gluconate, succinate, glutarate, adipate,
pimelate, suberate, azelate, sebacate, salicylate, phthalates,
aspartate, glutamate, and the like), and carboranes.
[0068] "Halo" or "halogen" refers to fluorine (fluoro, --F),
chlorine (chloro, --Cl), bromine (bromo, --Br), or iodine (iodo,
--I).
[0069] The term "hydroxyl" or "hydroxy" refers to the group --OH.
The term "substituted hydroxyl" or "substituted hydroxyl," by
extension, refers to a hydroxyl group wherein the oxygen atom
directly attached to the parent molecule is substituted with a
group other than hydrogen, and includes groups selected from
--OR.sup.aa, --ON(R.sup.bb).sub.2, --OC(.dbd.O)SR.sup.aa,
--OC(.dbd.O)R.sup.aa, --OCO.sub.2R.sup.aa,
--OC(.dbd.O)N(R.sup.bb).sub.2, --OC(.dbd.NR.sup.bb)R.sup.aa,
--OC(.dbd.NR.sup.bb)OR.sup.aa,
--OC(.dbd.NR.sup.bb)N(R.sup.bb).sub.2, --OS(.dbd.O)R.sup.aa,
--OSO.sub.2R.sup.aa, --OSi(R.sup.aa).sub.3, --OP(R.sup.cc).sub.2,
--OP(R.sup.cc).sub.3.sup.+X.sup.-, --OP(OR.sup.cc).sub.2,
--OP(OR.sup.cc).sub.3.sup.+X.sup.-, --OP(.dbd.O)(R.sup.aa).sub.2,
--OP(.dbd.O)(OR.sup.cc).sub.2, and --OP(.dbd.O)(N(R.sup.bb)).sub.2,
wherein X.sup.-, R.sup.aa, R.sup.bb, and R.sup.cc are as defined
herein.
[0070] "Acyl" refers to a moiety selected from the group consisting
of --C(.dbd.O)R.sup.aa, --CHO, --CO.sub.2R.sup.aa,
--C(.dbd.O)N(R.sup.bb).sub.2, --C(.dbd.NR.sup.bb)R.sup.aa,
--C(.dbd.NR.sup.bb)OR.sup.aa, --C(.dbd.NR.sup.bb)N(R.sup.bb).sub.2,
--C(.dbd.O)NR.sup.bbSO.sub.2R.sup.aa, --C(.dbd.S)N(R.sup.bb).sub.2,
--C(.dbd.O)SR.sup.aa, or --C(.dbd.S)SR.sup.aa, wherein R.sup.aa and
R.sup.bb are as defined herein.
[0071] The term "amino" refers to the group --NH.sub.2. The term
"substituted amino," by extension, refers to a monosubstituted
amino, a disubstituted amino, or a trisubstituted amino. In certain
embodiments, the "substituted amino" is a monosubstituted amino or
a disubstituted amino group.
[0072] Nitrogen atoms can be substituted or unsubstituted as
valency permits, and include primary, secondary, tertiary, and
quaternary nitrogen atoms. Exemplary nitrogen atom substituents
include, but are not limited to, hydrogen, --OH, --OR.sup.aa,
--N(R.sup.cc).sub.2, --CN, --C(.dbd.O)R.sup.aa,
--C(.dbd.O)N(R.sup.cc).sub.2, --CO.sub.2R.sup.aa,
--SO.sub.2R.sup.aa, --C(.dbd.NR.sup.bb)R.sup.aa,
--C(.dbd.NR.sup.cc)OR.sup.aa, --C(.dbd.NR.sup.cc)N(R.sup.cc).sub.2,
--SO.sub.2N(R.sup.cc).sub.2, --SO.sub.2R.sup.cc,
--SO.sub.2OR.sup.cc, --SOR.sup.aa, --C(.dbd.S)N(R.sup.cc).sub.2,
--C(.dbd.O)SR.sup.cc, --C(.dbd.S)SR.sup.cc,
--P(.dbd.O)(OR.sup.cc).sub.2, --P(.dbd.O)(R.sup.aa).sub.2,
--P(.dbd.O)(N(R.sup.cc).sub.2).sub.2, C.sub.1-10 alkyl, C.sub.1-10
perhaloalkyl, C.sub.2-10 alkenyl, C.sub.2-10 alkynyl,
heteroC.sub.1-10alkyl, heteroC.sub.2-10alkenyl,
heteroC.sub.2-10alkynyl, C.sub.3-10 carbocyclyl, 3-14 membered
heterocyclyl, C.sub.6-14 aryl, and 5-14 membered heteroaryl, or two
R.sup.cc groups attached to an N atom are joined to form a 3-14
membered heterocyclyl or 5-14 membered heteroaryl ring, wherein
each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl,
heteroalkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is
independently substituted with 0, 1, 2, 3, 4, or 5 R.sup.dd groups,
and wherein R.sup.aa, R.sup.bb, R.sup.cc and R.sup.dd are as
defined above.
[0073] In certain embodiments, the substituent present on a
nitrogen atom is a nitrogen protecting group (also referred to as
an amino protecting group). Nitrogen protecting groups include, but
are not limited to, --OH, --OR.sup.aa, --N(R.sup.cc).sub.2,
--C(.dbd.O)R.sup.aa, --C(.dbd.O)N(R.sup.cc).sub.2,
--CO.sub.2R.sup.aa, --SO.sub.2R.sup.aa,
--C(.dbd.NR.sup.cc)R.sup.aa, --C(.dbd.NR.sup.cc)OR.sup.aa,
--C(.dbd.NR.sup.cc)N(R.sup.cc).sub.2, --SO.sub.2N(R.sup.cc).sub.2,
--SO.sub.2R.sup.cc, --SO.sub.2OR.sup.cc, --SOR.sup.aa,
--C(.dbd.S)N(R.sup.cc).sub.2, --C(.dbd.O)SR.sup.cc,
--C(.dbd.S)SR.sup.cc, C.sub.1-10 alkyl (e.g., aralkyl,
heteroaralkyl), C.sub.2-10 alkenyl, C.sub.2-10 alkynyl, C.sub.3-10
carbocyclyl, 3-14 membered heterocyclyl, C.sub.6-14 aryl, and 5-14
membered heteroaryl groups, wherein each alkyl, alkenyl, alkynyl,
carbocyclyl, heterocyclyl, aralkyl, aryl, and heteroaryl is
independently substituted with 0, 1, 2, 3, 4, or 5 R.sup.dd groups,
and wherein R.sup.aa, R.sup.bb, R.sup.cc and R.sup.dd are as
defined herein. Nitrogen protecting groups are well known in the
art and include those described in detail in Protecting Groups in
Organic Synthesis, T. W. Greene and P. G. M. Wuts, 3.sup.rd
edition, John Wiley & Sons, 1999, incorporated herein by
reference.
[0074] For example, nitrogen protecting groups such as amide groups
(e.g., --C(.dbd.O)R.sup.aa) include, but are not limited to,
formamide, acetamide, chloroacetamide, trichloroacetamide,
trifluoroacetamide, phenylacetamide, 3-phenylpropanamide,
picolinamide, 3-pyridylcarboxamide, N-benzoylphenylalanyl
derivative, benzamide, p-phenylbenzamide, o-nitrophenylacetamide,
o-nitrophenoxyacetamide, acetoacetamide,
(N'-dithiobenzyloxyacylamino)acetamide,
3-(p-hydroxyphenyl)propanamide, 3-(o-nitrophenyl)propanamide,
2-methyl-2-(o-nitrophenoxy)propanamide,
2-methyl-2-(o-phenylazophenoxy)propanamide, 4-chlorobutanamide,
3-methyl-3-nitrobutanamide, o-nitrocinnamide, N-acetylmethionine
derivative, o-nitrobenzamide, and
o-(benzoyloxymethyl)benzamide.
[0075] Nitrogen protecting groups such as carbamate groups (e.g.,
--C(.dbd.O)OR.sup.aa) include, but are not limited to, methyl
carbamate, ethyl carbamate, 9-fluorenylmethyl carbamate (Fmoc),
9-(2-sulfo)fluorenylmethyl carbamate,
9-(2,7-dibromo)fluorenylmethyl carbamate,
2,7-di-t-butyl-[9-(10,10-dioxo-10,10,10,10-tetrahydrothioxanthyl)]methyl
carbamate (DBD-Tmoc), 4-methoxyphenacyl carbamate (Phenoc),
2,2,2-trichloroethyl carbamate (Troc), 2-trimethylsilylethyl
carbamate (Teoc), 2-phenylethyl carbamate (hZ),
1-(1-adamantyl)-1-methylethyl carbamate (Adpoc),
1,1-dimethyl-2-haloethyl carbamate, 1,1-dimethyl-2,2-dibromoethyl
carbamate (DB-t-BOC), 1,1-dimethyl-2,2,2-trichloroethyl carbamate
(TCBOC), 1-methyl-1-(4-biphenylyl)ethyl carbamate (Bpoc),
1-(3,5-di-t-butylphenyl)-1-methylethyl carbamate (t-Bumeoc), 2-(2'-
and 4'-pyridyl)ethyl carbamate (Pyoc),
2-(N,N-dicyclohexylcarboxamido)ethyl carbamate, t-butyl carbamate
(BOC or Boc), 1-adamantyl carbamate (Adoc), vinyl carbamate (Voc),
allyl carbamate (Alloc), 1-isopropylallyl carbamate (Ipaoc),
cinnamyl carbamate (Coc), 4-nitrocinnamyl carbamate (Noc),
8-quinolyl carbamate, N-hydroxypiperidinyl carbamate, alkyldithio
carbamate, benzyl carbamate (Cbz), p-methoxybenzyl carbamate (Moz),
p-nitrobenzyl carbamate, p-bromobenzyl carbamate, p-chlorobenzyl
carbamate, 2,4-dichlorobenzyl carbamate, 4-methylsulfinylbenzyl
carbamate (Msz), 9-anthrylmethyl carbamate, diphenylmethyl
carbamate, 2-methylthioethyl carbamate, 2-methylsulfonylethyl
carbamate, 2-(p-toluenesulfonyl)ethyl carbamate,
[2-(1,3-dithianyl)]methyl carbamate (Dmoc), 4-methylthiophenyl
carbamate (Mtpc), 2,4-dimethylthiophenyl carbamate (Bmpc),
2-phosphonioethyl carbamate (Peoc), 2-triphenylphosphonioisopropyl
carbamate (Ppoc), 1,1-dimethyl-2-cyanoethyl carbamate,
m-chloro-p-acyloxybenzyl carbamate, p-(dihydroxyboryl)benzyl
carbamate, 5-benzisoxazolylmethyl carbamate,
2-(trifluoromethyl)-6-chromonylmethyl carbamate (Tcroc),
m-nitrophenyl carbamate, 3,5-dimethoxybenzyl carbamate,
o-nitrobenzyl carbamate, 3,4-dimethoxy-6-nitrobenzyl carbamate,
phenyl(o-nitrophenyl)methyl carbamate, t-amyl carbamate, S-benzyl
thiocarbamate, p-cyanobenzyl carbamate, cyclobutyl carbamate,
cyclohexyl carbamate, cyclopentyl carbamate, cyclopropylmethyl
carbamate, p-decyloxybenzyl carbamate, 2,2-dimethoxyacylvinyl
carbamate, o-(N,N-dimethylcarboxamido)benzyl carbamate,
1,1-dimethyl-3-(N,N-dimethylcarboxamido)propyl carbamate,
1,1-dimethylpropynyl carbamate, di(2-pyridyl)methyl carbamate,
2-furanylmethyl carbamate, 2-iodoethyl carbamate, isoborynl
carbamate, isobutyl carbamate, isonicotinyl carbamate,
p-(p'-methoxyphenylazo)benzyl carbamate, 1-methylcyclobutyl
carbamate, 1-methylcyclohexyl carbamate,
1-methyl-1-cyclopropylmethyl carbamate,
1-methyl-1-(3,5-dimethoxyphenyl)ethyl carbamate,
i-methyl-1-(p-phenylazophenyl)ethyl carbamate,
1-methyl-1-phenylethyl carbamate, 1-methyl-1-(4-pyridyl)ethyl
carbamate, phenyl carbamate, p-(phenylazo)benzyl carbamate,
2,4,6-tri-t-butylphenyl carbamate, 4-(trimethylammonium)benzyl
carbamate, and 2,4,6-trimethylbenzyl carbamate.
[0076] Nitrogen protecting groups such as sulfonamide groups (e.g.,
--S(.dbd.O).sub.2R.sup.aa) include, but are not limited to,
p-toluenesulfonamide (Ts), benzenesulfonamide,
2,3,6,-trimethyl-4-methoxybenzenesulfonamide (Mtr),
2,4,6-trimethoxybenzenesulfonamide (Mtb),
2,6-dimethyl-4-methoxybenzenesulfonamide (Pme),
2,3,5,6-tetramethyl-4-methoxybenzenesulfonamide (Mte),
4-methoxybenzenesulfonamide (Mbs),
2,4,6-trimethylbenzenesulfonamide (Mts),
2,6-dimethoxy-4-methylbenzenesulfonamide (iMds),
2,2,5,7,8-pentamethylchroman-6-sulfonamide (Pmc),
methanesulfonamide (Ms), .beta.-trimethylsilylethanesulfonamide
(SES), 9-anthracenesulfonamide,
4-(4',8'-dimethoxynaphthylmethyl)benzenesulfonamide (DNMBS),
benzylsulfonamide, trifluoromethylsulfonamide, and
phenacylsulfonamide.
[0077] Other nitrogen protecting groups include, but are not
limited to, phenothiazinyl-(10)-acyl derivative,
N'-p-toluenesulfonylaminoacyl derivative, N'-phenylaminothioacyl
derivative, N-benzoylphenylalanyl derivative, N-acetylmethionine
derivative, 4,5-diphenyl-3-oxazolin-2-one, N-phthalimide,
N-dithiasuccinimide (Dts), N-2,3-diphenylmaleimide,
N-2,5-dimethylpyrrole, N-1,1,4,4-tetramethyldisilylazacyclopentane
adduct (STABASE), 5-substituted
1,3-dimethyl-1,3,5-triazacyclohexan-2-one, 5-substituted
1,3-dibenzyl-1,3,5-triazacyclohexan-2-one, 1-substituted
3,5-dinitro-4-pyridone, N-methylamine, N-allylamine,
N-[2-(trimethylsilyl)ethoxy]methylamine (SEM),
N-3-acetoxypropylamine,
N-(1-isopropyl-4-nitro-2-oxo-3-pyroolin-3-yl)amine, quaternary
ammonium salts, N-benzylamine, N-di(4-methoxyphenyl)methylamine,
N-5-dibenzosuberylamine, N-triphenylmethylamine (Tr),
N-[(4-methoxyphenyl)diphenylmethyl]amine (MMTr),
N-9-phenylfluorenylamine (PhF),
N-2,7-dichloro-9-fluorenylmethyleneamine, N-ferrocenylmethylamino
(Fcm), N-2-picolylamino N-oxide, N-1,1-dimethylthiomethyleneamine,
N-benzylideneamine, N-p-methoxybenzylideneamine,
N-diphenylmethyleneamine, N-[(2-pyridyl)mesityl]methyleneamine,
N--(N',N'-dimethylaminomethylene)amine, N,N'-isopropylidenediamine,
N-p-nitrobenzylideneamine, N-salicylideneamine,
N-5-chlorosalicylideneamine,
N-(5-chloro-2-hydroxyphenyl)phenylmethyleneamine,
N-cyclohexylideneamine, N-(5,5-dimethyl-3-oxo-1-cyclohexenyl)amine,
N-borane derivative, N-diphenylborinic acid derivative,
N-[phenyl(pentaacylchromium- or tungsten)acyl]amine, N-copper
chelate, N-zinc chelate, N-nitroamine, N-nitrosoamine, amine
N-oxide, diphenylphosphinamide (Dpp), dimethylthiophosphinamide
(Mpt), diphenylthiophosphinamide (Ppt), dialkyl phosphoramidates,
dibenzyl phosphoramidate, diphenyl phosphoramidate,
benzenesulfenamide, o-nitrobenzenesulfenamide (Nps),
2,4-dinitrobenzenesulfenamide, pentachlorobenzenesulfenamide,
2-nitro-4-methoxybenzenesulfenamide, triphenylmethylsulfenamide,
and 3-nitropyridinesulfenamide (Npys).
[0078] Exemplary oxygen atom substituents include, but are not
limited to, --R.sup.aa, --N(R.sup.bb).sub.2, --C(.dbd.O)SR.sup.aa,
--C(.dbd.O)R.sup.aa, --CO.sub.2R.sup.aa,
--C(.dbd.O)N(R.sup.bb).sub.22, --C(.dbd.NR.sup.bb)R.sup.aa,
--C(.dbd.NR.sup.bb)OR.sup.aa, --C(.dbd.NR.sup.bb)N(R.sup.bb),
--S(.dbd.O)R.sup.aa, --SO.sub.2R.sup.aa, --Si(R.sup.aa).sub.3,
--P(R.sup.cc).sub.2, --P(R.sup.cc).sub.3.sup.+X.sup.-,
--P(OR.sup.cc).sub.2, --P(OR.sup.cc).sub.3.sup.+X.sup.-,
--P(.dbd.O)(R.sup.aa).sub.2, --P(.dbd.O)(OR.sup.cc).sub.2, and
--P(.dbd.O)(N(R.sup.bb).sub.2).sub.2, wherein X.sup.-, R.sup.aa,
R.sup.bb, and R.sup.cc are as defined herein. In certain
embodiments, the oxygen atom substituent present on an oxygen atom
is an oxygen protecting group (also referred to as a hydroxyl
protecting group). Oxygen protecting groups are well known in the
art and include those described in detail in Protecting Groups in
Organic Synthesis, T. W. Greene and P. G. M. Wuts, 3.sup.rd
edition, John Wiley & Sons, 1999, incorporated herein by
reference. Exemplary oxygen protecting groups include, but are not
limited to, methyl, t-butyloxycarbonyl (BOC or Boc), methoxylmethyl
(MOM), methylthiomethyl (MTM), t-butylthiomethyl,
(phenyldimethylsilyl)methoxymethyl (SMOM), benzyloxymethyl (BOM),
p-methoxybenzyloxymethyl (PMBM), (4-methoxyphenoxy)methyl (p-AOM),
guaiacolmethyl (GUM), t-butoxymethyl, 4-pentenyloxymethyl (POM),
siloxymethyl, 2-methoxyethoxymethyl (MEM),
2,2,2-trichloroethoxymethyl, bis(2-chloroethoxy)methyl,
2-(trimethylsilyl)ethoxymethyl (SEMOR), tetrahydropyranyl (THP),
3-bromotetrahydropyranyl, tetrahydrothiopyranyl,
1-methoxycyclohexyl, 4-methoxytetrahydropyranyl (MTHP),
4-methoxytetrahydrothiopyranyl, 4-methoxytetrahydrothiopyranyl
S,S-dioxide, 1-[(2-chloro-4-methyl)phenyl]-4-methoxypiperidin-4-yl
(CTMP), 1,4-dioxan-2-yl, tetrahydrofuranyl, tetrahydrothiofuranyl,
2,3,3a,4,5,6,7,7a-octahydro-7,8,8-trimethyl-4,7-methanobenzofuran-2-yl,
1-ethoxyethyl, 1-(2-chloroethoxy)ethyl, 1-methyl-1-methoxyethyl,
1-methyl-1-benzyloxyethyl, 1-methyl-1-benzyloxy-2-fluoroethyl,
2,2,2-trichloroethyl, 2-trimethylsilylethyl,
2-(phenylselenyl)ethyl, t-butyl, allyl, p-chlorophenyl,
p-methoxyphenyl, 2,4-dinitrophenyl, benzyl (Bn), p-methoxybenzyl,
3,4-dimethoxybenzyl, o-nitrobenzyl, p-nitrobenzyl, p-halobenzyl,
2,6-dichlorobenzyl, p-cyanobenzyl, p-phenylbenzyl, 2-picolyl,
4-picolyl, 3-methyl-2-picolyl N-oxido, diphenylmethyl,
p,p'-dinitrobenzhydryl, 5-dibenzosuberyl, triphenylmethyl,
.alpha.-naphthyldiphenylmethyl, p-methoxyphenyldiphenylmethyl,
di(p-methoxyphenyl)phenylmethyl, tri(p-methoxyphenyl)methyl,
4-(4'-bromophenacyloxyphenyl)diphenylmethyl,
4,4',4''-tris(4,5-dichlorophthalimidophenyl)methyl,
4,4',4''-tris(levulinoyloxyphenyl)methyl,
4,4',4''-tris(benzoyloxyphenyl)methyl,
3-(imidazol-1-yl)bis(4',4''-dimethoxyphenyl)methyl,
1,1-bis(4-methoxyphenyl)-1'-pyrenylmethyl, 9-anthryl,
9-(9-phenyl)xanthenyl, 9-(9-phenyl-10-oxo)anthryl,
1,3-benzodisulfuran-2-yl, benzisothiazolyl S,S-dioxido,
trimethylsilyl (TMS), triethylsilyl (TES), triisopropylsilyl
(TIPS), dimethylisopropylsilyl (IPDMS), diethylisopropylsilyl
(DEIPS), dimethylthexylsilyl, I-butyldimethylsilyl (TBDMS),
t-butyldiphenylsilyl (TBDPS), tribenzylsilyl, tri-p-xylylsilyl,
triphenylsilyl, diphenylmethylsilyl (DPMS),
t-butylmethoxyphenylsilyl (TBMPS), formate, benzoylformate,
acetate, chloroacetate, dichloroacetate, trichloroacetate,
trifluoroacetate, methoxyacetate, triphenylmethoxyacetate,
phenoxyacetate, p-chlorophenoxyacetate, 3-phenylpropionate,
4-oxopentanoate (levulinate), 4,4-(ethylenedithio)pentanoate
(levulinoyldithioacetal), pivaloate, adamantoate, crotonate,
4-methoxycrotonate, benzoate, p-phenylbenzoate,
2,4,6-trimethylbenzoate (mesitoate), alkyl methyl carbonate,
9-fluorenylmethyl carbonate (Fmoc), alkyl ethyl carbonate, alkyl
2,2,2-trichloroethyl carbonate (Troc), 2-(trimethylsilyl)ethyl
carbonate (TMSEC), 2-(phenylsulfonyl) ethyl carbonate (Psec),
2-(triphenylphosphonio) ethyl carbonate (Peoc), alkyl isobutyl
carbonate, alkyl vinyl carbonate alkyl allyl carbonate, alkyl
p-nitrophenyl carbonate, alkyl benzyl carbonate, alkyl
p-methoxybenzyl carbonate, alkyl 3,4-dimethoxybenzyl carbonate,
alkyl o-nitrobenzyl carbonate, alkyl p-nitrobenzyl carbonate, alkyl
S-benzyl thiocarbonate, 4-ethoxy-1-naphthyl carbonate, methyl
dithiocarbonate, 2-iodobenzoate, 4-azidobutyrate,
4-nitro-4-methylpentanoate, o-(dibromomethyl)benzoate,
2-formylbenzenesulfonate, 2-(methylthiomethoxy)ethyl,
4-(methylthiomethoxy)butyrate, 2-(methylthiomethoxymethyl)benzoate,
2,6-dichloro-4-methylphenoxyacetate,
2,6-dichloro-4-(1,1,3,3-tetramethylbutyl)phenoxyacetate,
2,4-bis(1,1-dimethylpropyl)phenoxyacetate, chlorodiphenylacetate,
isobutyrate, monosuccinoate, (E)-2-methyl-2-butenoate,
o-(methoxyacyl)benzoate, .alpha.-naphthoate, nitrate, alkyl
N,N,N',N'-tetramethylphosphorodiamidate, alkyl N-phenylcarbamate,
borate, dimethylphosphinothioyl, alkyl 2,4-dinitrophenylsulfenate,
sulfate, methanesulfonate (mesylate), benzylsulfonate, and tosylate
(Ts).
[0079] Exemplary sulfur atom substituents include, but are not
limited to, --R.sup.aa, --N(R.sup.bb).sub.2, --C(.dbd.O)SR.sup.aa,
--C(.dbd.O)R.sup.aa, --CO.sub.2R.sup.aa,
--C(.dbd.O)N(R.sup.bb).sub.2, --C(.dbd.NR.sup.bb)R.sup.aa,
--C(.dbd.NR.sup.bb)OR.sup.aa, --C(.dbd.NR.sup.bb)N(R.sup.bb).sub.2,
--S(.dbd.O)R.sup.aa, --SO.sub.2R.sup.aa, --Si(R.sup.aa).sub.3,
--P(R.sup.cc).sub.2, --P(R.sup.cc).sub.3.sup.+X.sup.-,
--P(OR.sup.cc).sub.2, --P(OR.sup.cc).sub.3.sup.+X.sup.-,
--P(.dbd.O)(R.sup.aa).sub.2, --P(.dbd.O)(OR.sup.cc).sub.2, and
--P(.dbd.O)(N(R.sup.bb).sub.2).sub.2, wherein R.sup.aa, R.sup.bb,
and R.sup.cc are as defined herein. In certain embodiments, the
sulfur atom substituent present on a sulfur atom is a sulfur
protecting group (also referred to as a thiol protecting group).
Sulfur protecting groups are well known in the art and include
those described in detail in Protecting Groups in Organic
Synthesis, T. W. Greene and P. G. M. Wuts, 3.sup.rd edition, John
Wiley & Sons, 1999, incorporated herein by reference.
[0080] A "hydrocarbon chain" refers to a substituted or
unsubstituted divalent alkyl, alkenyl, or alkynyl group. A
hydrocarbon chain includes (1) one or more chains of carbon atoms
immediately between the two radicals of the hydrocarbon chain; (2)
optionally one or more hydrogen atoms on the chain(s) of carbon
atoms; and (3) optionally one or more substituents ("non-chain
substituents," which are not hydrogen) on the chain(s) of carbon
atoms. A chain of carbon atoms consists of consecutively connected
carbon atoms ("chain atoms" or "carbon units") and does not include
hydrogen atoms or heteroatoms. However, a non-chain substituent of
a hydrocarbon chain may include any atoms, including hydrogen
atoms, carbon atoms, and heteroatoms. For example, hydrocarbon
chain --C.sup.AH(C.sup.BH.sub.2C.sup.CH.sub.3)-- includes one chain
atom C.sup.A, one hydrogen atom on C.sup.A, and non-chain
substituent --(C.sup.BH.sub.2C.sup.CH.sub.3). The term "C.sub.x
hydrocarbon chain," wherein x is a positive integer, refers to a
hydrocarbon chain that includes x number of chain atom(s) between
the two radicals of the hydrocarbon chain. If there is more than
one possible value of x, the smallest possible value of x is used
for the definition of the hydrocarbon chain. For example,
--CH(C.sub.2H.sub.5)-- is a C.sub.1 hydrocarbon chain, and
##STR00006##
is a C.sub.3 hydrocarbon chain. When a range of values is used, the
meaning of the range is as described herein. For example, a
C.sub.3-10 hydrocarbon chain refers to a hydrocarbon chain where
the number of chain atoms of the shortest chain of carbon atoms
immediately between the two radicals of the hydrocarbon chain is 3,
4, 5, 6, 7, 8, 9, or 10. A hydrocarbon chain may be saturated
(e.g., --(CH.sub.2).sub.4--). A hydrocarbon chain may also be
unsaturated and include one or more C.dbd.C and/or C.ident.C bonds
anywhere in the hydrocarbon chain. For instance,
--CH.dbd.CH--(CH.sub.2).sub.2--, --CH.sub.2--C.ident.C--CH.sub.2--,
and --C.ident.C--CH.dbd.CH-- are all examples of a unsubstituted
and unsaturated hydrocarbon chain. In certain embodiments, the
hydrocarbon chain is unsubstituted (e.g., --C.ident.C-- or
--(CH.sub.2).sub.4--). In certain embodiments, the hydrocarbon
chain is substituted (e.g., --CH(C.sub.2H.sub.5)-- and
--CF.sub.2--). Any two substituents on the hydrocarbon chain may be
joined to form an optionally substituted carbocyclyl, optionally
substituted heterocyclyl, optionally substituted aryl, or
optionally substituted heteroaryl ring. For instance,
##STR00007##
are all examples of a hydrocarbon chain. In contrast, in certain
embodiments,
##STR00008##
are not within the scope of the hydrocarbon chains described
herein. When a chain atom of a C.sub.x hydrocarbon chain is
replaced with a heteroatom, the resulting group is referred to as a
C.sub.x hydrocarbon chain wherein a chain atom is replaced with a
heteroatom, as opposed to a C.sub.x-1 hydrocarbon chain. For
example,
##STR00009##
is a C.sub.3 hydrocarbon chain wherein one chain atom is replaced
with an oxygen atom.
[0081] The term "leaving group" is given its ordinary meaning in
the art of synthetic organic chemistry and refers to an atom or a
group capable of being displaced by a nucleophile. Examples of
suitable leaving groups include, but are not limited to, halogen
(such as F, Cl, Br, or I (iodine)), alkoxycarbonyloxy,
aryloxycarbonyloxy, alkanesulfonyloxy, arenesulfonyloxy,
alkyl-carbonyloxy (e.g., acetoxy), arylcarbonyloxy, aryloxy,
methoxy, N,O-dimethylhydroxylamino, pixel, and haloformates. In
some cases, the leaving group is a sulfonic acid ester, such as
toluenesulfonate (tosylate, --OTs), methanesulfonate (mesylate,
--OMs), p-bromobenzenesulfonyloxy (brosylate, --OBs),
--OS(.dbd.O).sub.2(CF.sub.2).sub.3CF.sub.3 (nonaflate, --ONf), or
trifluoromethanesulfonate (triflate, -OTf). In some cases, the
leaving group is a brosylate, such as p-bromobenzenesulfonyloxy. In
some cases, the leaving group is a nosylate, such as
2-nitrobenzenesulfonyloxy. The leaving group may also be a
phosphineoxide (e.g., formed during a Mitsunobu reaction) or an
internal leaving group such as an epoxide or cyclic sulfate. Other
non-limiting examples of leaving groups are water, ammonia,
alcohols, ether moieties, thioether moieties, zinc halides,
magnesium moieties, diazonium salts, and copper moieties. Exemplary
leaving groups include, but are not limited to, halo (e.g., chloro,
bromo, iodo) and activated substituted hydroxyl groups (e.g.,
--OC(.dbd.O)SR.sup.aa, --OC(.dbd.O)R.sup.aa, --OCO.sub.2R.sup.aa,
--OC(.dbd.O)N(R.sup.bb).sub.2, --OC(.dbd.NR.sup.bb)R.sup.aa,
--OC(.dbd.NR.sup.bb)OR.sup.aa,
--OC(.dbd.NR.sup.bb)N(R.sup.bb).sub.2, --OS(.dbd.O)R.sup.aa,
--OSO.sub.2R.sup.aa, --OP(R.sup.cc).sub.2, --OP(R.sup.cc).sub.3,
--OP(.dbd.O).sub.2R.sup.aa, --OP(.dbd.O)(R.sup.aa).sub.2,
--OP(.dbd.O)(OR.sup.cc).sub.2, --OP(.dbd.O).sub.2N(R.sup.bb).sub.2,
and --OP(.dbd.O)(NR.sup.bb).sub.2, wherein R.sup.aa, R.sup.bb, and
R.sup.cc are as defined herein).
[0082] The term "pharmaceutically acceptable salt" refers to those
salts which are, within the scope of sound medical judgment,
suitable for use in contact with the tissues of humans and lower
animals without undue toxicity, irritation, allergic response, and
the like, and are commensurate with a reasonable benefit/risk
ratio. Pharmaceutically acceptable salts are well known in the art.
For example, Berge et al. describe pharmaceutically acceptable
salts in detail in J. Pharmaceutical Sciences, 1977, 66, 1-19,
incorporated herein by reference. Pharmaceutically acceptable salts
of the compounds described herein include those derived from
suitable inorganic and organic acids and bases. Examples of
pharmaceutically acceptable, non-toxic acid addition salts are
salts of an amino group formed with inorganic acids such as
hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric
acid, and perchloric acid or with organic acids such as acetic
acid, oxalic acid, maleic acid, tartaric acid, citric acid,
succinic acid, or malonic acid or by using other methods known in
the art such as ion exchange. Other pharmaceutically acceptable
salts include adipate, alginate, ascorbate, aspartate,
benzenesulfonate, benzoate, bisulfate, borate, butyrate,
camphorate, camphorsulfonate, citrate, cyclopentanepropionate,
digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate,
glucoheptonate, glycerophosphate, gluconate, hemisulfate,
heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate,
lactobionate, lactate, laurate, lauryl sulfate, malate, maleate,
malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate,
nitrate, oleate, oxalate, palmitate, pamoate, pectinate,
persulfate, 3-phenylpropionate, phosphate, picrate, pivalate,
propionate, stearate, succinate, sulfate, tartrate, thiocyanate,
p-toluenesulfonate, undecanoate, valerate salts, and the like.
Salts derived from appropriate bases include alkali metal, alkaline
earth metal, ammonium and N.sup.+(C.sub.1-4 alkyl).sub.4.sup.-
salts. Representative alkali or alkaline earth metal salts include
sodium, lithium, potassium, calcium, magnesium, and the like.
Further pharmaceutically acceptable salts include, when
appropriate, nontoxic ammonium, quaternary ammonium, and amine
cations formed using counterions such as halide, hydroxide,
carboxylate, sulfate, phosphate, nitrate, lower alkyl sulfonate,
and aryl sulfonate.
[0083] The term "solvate" refers to forms of the compound that are
associated with a solvent, usually by a solvolysis reaction. This
physical association may include hydrogen bonding. Conventional
solvents include water, methanol, ethanol, acetic acid, DMSO, THF,
diethyl ether, and the like. The compounds described herein may be
prepared, e.g., in crystalline form, and may be solvated. Suitable
solvates include pharmaceutically acceptable solvates and further
include both stoichiometric solvates and non-stoichiometric
solvates. In certain instances, the solvate will be capable of
isolation, for example, when one or more solvent molecules are
incorporated in the crystal lattice of a crystalline solid.
"Solvate" encompasses both solution-phase and isolatable solvates.
Representative solvates include hydrates, ethanolates, and
methanolates.
[0084] The term "hydrate" refers to a compound that is associated
with water. Typically, the number of the water molecules contained
in a hydrate of a compound is in a definite ratio to the number of
the compound molecules in the hydrate. Therefore, a hydrate of a
compound may be represented, for example, by the general formula
R.x H.sub.2O, wherein R is the compound, and x is a number greater
than 0. A given compound may form more than one type of hydrate,
including, e.g., monohydrates (x is 1), lower hydrates (x is a
number greater than 0 and smaller than 1, e.g., hemihydrates
(R.0.5H.sub.2O)), and polyhydrates (x is a number greater than 1,
e.g., dihydrates (R.2H.sub.2O) and hexahydrates (R.6H.sub.2O)).
[0085] The term "tautomers" or "tautomeric" refers to two or more
interconvertible compounds resulting from at least one formal
migration of a hydrogen atom and at least one change in valency
(e.g., a single bond to a double bond, a triple bond to a single
bond, or vice versa). The exact ratio of the tautomers depends on
several factors, including temperature, solvent, and pH.
Tautomerizations (i.e., the reaction providing a tautomeric pair)
may catalyzed by acid or base. Exemplary tautomerizations include
keto-to-enol, amide-to-imide, lactam-to-lactim, enamine-to-imine,
and enamine-to-(a different enamine) tautomerizations.
[0086] It is also to be understood that compounds that have the
same molecular formula but differ in the nature or sequence of
bonding of their atoms or the arrangement of their atoms in space
are termed "isomers". Isomers that differ in the arrangement of
their atoms in space are termed "stereoisomers."
[0087] Stereoisomers that are not mirror images of one another are
termed "diastereomers" and those that are non-superimposable mirror
images of each other are termed "enantiomers." When a compound has
an asymmetric center, for example, it is bonded to four different
groups, a pair of enantiomers is possible. An enantiomer can be
characterized by the absolute configuration of its asymmetric
center and is described by the R- and S-sequencing rules of Cahn
and Prelog, or by the manner in which the molecule rotates the
plane of polarized light and designated as dextrorotatory or
levorotatory (i.e., as (+) or (-)-isomers respectively). A chiral
compound can exist as either individual enantiomer or as a mixture
thereof. A mixture containing equal proportions of the enantiomers
is called a "racemic mixture."
[0088] The term "polymorphs" refers to a crystalline form of a
compound (or a salt, hydrate, or solvate thereof) in a particular
crystal packing arrangement. All polymorphs have the same elemental
composition. Different crystalline forms usually have different
X-ray diffraction patterns, infrared spectra, melting points,
density, hardness, crystal shape, optical and electrical
properties, stability, and solubility. Recrystallization solvent,
rate of crystallization, storage temperature, and other factors may
cause one crystal form to dominate. Various polymorphs of a
compound can be prepared by crystallization under different
conditions.
[0089] The term "co-crystal" refers to a crystalline structure
composed of at least two components. In certain embodiments, a
co-crystal may contain a compound of the present invention and one
or more other component, including but not limited to, atoms, ions,
molecules, or solvent molecules. In certain embodiments, a
co-crystal may contain a compound of the present invention and one
or more components related to said compound, including not limited
to, an isomer, tautomer, salt, solvate, hydrate, synthetic
precursor, synthetic derivative, fragment or impurity of said
compound.
[0090] The term "isotopically labeled derivative" or "isotopically
labeled" refers to a compound wherein one or more atoms in the
compound (or in an associated ion or molecule of a salt, hydrate,
or solvate) has been replaced with an isotope of the same element.
For the given element or position in the molecule the isotope will
be enriched, or present in a higher percentage of all atoms of the
element or of all atoms at the position in the molecule in a
sample, relative to an unlabeled variant. In certain embodiments,
the enriched isotope will be a stable isotope. In certain
embodiments, the enriched isotope will be an unstable or
radioactive isotope (e.g., a radionuclide). In certain embodiments,
the enriched isotope may be detected by a measurement technique,
including but not limited to nuclear magnetic resonance, mass
spectrometry, infrared spectroscopy, or a technique that measures
radioactive decay.
[0091] The term "prodrugs" refers to compounds that have cleavable
groups and become by solvolysis or under physiological conditions
the compounds described herein, which are pharmaceutically active
in vivo. Such examples include, but are not limited to, choline
ester derivatives and the like, N-alkylmorpholine esters and the
like. Other derivatives of the compounds described herein have
activity in both their acid and acid derivative forms, but in the
acid sensitive form often offer advantages of solubility, tissue
compatibility, or delayed release in the mammalian organism (see,
Bundgard, H., Design of Prodrugs, pp. 7-9, 21-24, Elsevier,
Amsterdam 1985). Prodrugs include acid derivatives well known to
practitioners of the art, such as, for example, esters prepared by
reaction of the parent acid with a suitable alcohol, or amides
prepared by reaction of the parent acid compound with a substituted
or unsubstituted amine, or acid anhydrides, or mixed anhydrides.
Simple aliphatic or aromatic esters, amides, and anhydrides derived
from acidic groups pendant on the compounds described herein are
particular prodrugs. In some cases it is desirable to prepare
double ester type prodrugs such as (acyloxy)alkyl esters or
((alkoxycarbonyl)oxy)alkylesters. C.sub.1-C.sub.8 alkyl,
C.sub.2-C.sub.8 alkenyl, C.sub.2-C.sub.8 alkynyl, aryl,
C.sub.7-C.sub.12 substituted aryl, and C.sub.7-C.sub.12 arylalkyl
esters of the compounds described herein may be preferred.
[0092] The term "inhibition", "inhibiting", "inhibit," or
"inhibitor" refer to the ability of a compound to reduce, slow,
halt or prevent activity of a particular biological process (e.g.,
CDK kinase activity)) in a cell relative to vehicle.
[0093] When a compound, pharmaceutical composition, method, use, or
kit is referred to as "selectively," "specifically," or
"competitively" binding a first protein kinase, the compound,
pharmaceutical composition, method, use, or kit binds the first
protein kinase (e.g., CDK) with a higher binding affinity (e.g.,
not less than about 2-fold, not less than about 5-fold, not less
than about 10-fold, not less than about 30-fold, not less than
about 100-fold, not less than about 1,000-fold, or not less than
about 10,000-fold) than binding a second protein or second
chromatin that is different from the first protein and the first
chromatin.
[0094] When a compound, pharmaceutical composition, method, use, or
kit is referred to as "selectively," "specifically," or
"competitively" modulating (e.g., increasing or inhibiting) the
activity of a first protein kinase, the compound, pharmaceutical
composition, method, use, or kit modulates the activity of the
first protein kinase (e.g., CDK) to a greater extent (e.g., not
less than about 2-fold, not less than about 5-fold, not less than
about 10-fold, not less than about 30-fold, not less than about
100-fold, not less than about 1,000-fold, or not less than about
10,000-fold) than the activity of a second protein kinase that is
different from the first protein kinase.
[0095] The term "aberrant activity" refers to activity deviating
from normal activity, that is, abnormal activity. The term
"increased activity" refers to activity higher than normal
activity.
[0096] The terms "composition" and "formulation" are used
interchangeably.
[0097] A "subject" to which administration is contemplated refers
to a human (i.e., male or female of any age group, e.g., pediatric
subject (e.g., infant, child, or adolescent) or adult subject
(e.g., young adult, middle-aged adult, or senior adult)) or
non-human animal. In certain embodiments, the non-human animal is a
mammal (e.g., primate (e.g., cynomolgus monkey or rhesus monkey),
commercially relevant mammal (e.g., cattle, pig, horse, sheep,
goat, cat, or dog), or bird (e.g., commercially relevant bird, such
as chicken, duck, goose, or turkey)). In certain embodiments, the
non-human animal is a fish, reptile, or amphibian. The non-human
animal may be a male or female at any stage of development. The
non-human animal may be a transgenic animal or genetically
engineered animal. A "patient" refers to a human subject in need of
treatment of a disease.
[0098] The term "biological sample" refers to any sample including
tissue samples (such as tissue sections and needle biopsies of a
tissue); cell samples (e.g., cytological smears (such as Pap or
blood smears) or samples of cells obtained by microdissection);
samples of whole organisms (such as samples of yeasts or bacteria);
or cell fractions, fragments or organelles (such as obtained by
lysing cells and separating the components thereof by
centrifugation or otherwise). Other examples of biological samples
include blood, serum, urine, semen, fecal matter, cerebrospinal
fluid, interstitial fluid, mucous, tears, sweat, pus, biopsied
tissue (e.g., obtained by a surgical biopsy or needle biopsy),
nipple aspirates, milk, vaginal fluid, saliva, swabs (such as
buccal swabs), or any material containing biomolecules that is
derived from another biological sample.
[0099] The terms "administer," "administering," or "administration"
refers to implanting, absorbing, ingesting, injecting, inhaling, or
otherwise introducing a compound described herein, or a composition
thereof, into, in, or on a subject.
[0100] The terms "treatment," "treat," and "treating" refer to
reversing, alleviating, delaying the onset of or inhibiting the
progress of a disease described herein. In some embodiments,
treatment may be administered after one or more signs or symptoms
of the disease have developed or have been observed. In other
embodiments, treatment may be administered in the absence of signs
or symptoms of the disease. For example, treatment may be
administered to a susceptible subject prior to the onset of
symptoms (e.g., in light of a history of symptoms and/or in light
of exposure to a pathogen). Treatment may also be continued after
symptoms have resolved, for example, to delay or prevent
recurrence.
[0101] The terms "condition," "disease," and "disorder" are used
interchangeably.
[0102] An "effective amount" of a compound described herein refers
to an amount sufficient to elicit the desired biological response,
i.e., treating the condition. As will be appreciated by those of
ordinary skill in this art, the effective amount of a compound
described herein may vary depending on such factors as the desired
biological endpoint, the pharmacokinetics of the compound, the
condition being treated, the mode of administration, and the age
and health of the subject. In certain embodiments, an effective
amount is a therapeutically effective amount. In certain
embodiments, an effective amount is a prophylactic treatment. In
certain embodiments, an effective amount is the amount of a
compound described herein in a single dose. In certain embodiments,
an effective amount is the combined amounts of a compound described
herein in multiple doses.
[0103] A "therapeutically effective amount" of a compound described
herein is an amount sufficient to provide a therapeutic benefit in
the treatment of a condition or to delay or minimize one or more
symptoms associated with the condition. A therapeutically effective
amount of a compound means an amount of therapeutic agent, alone or
in combination with other therapies, which provides a therapeutic
benefit in the treatment of the condition. The term
"therapeutically effective amount" can encompass an amount that
improves overall therapy, reduces or avoids symptoms, signs, or
causes of the condition, and/or enhances the therapeutic efficacy
of another therapeutic agent.
[0104] A "prophylactically effective amount" of a compound
described herein is an amount sufficient to prevent a condition, or
one or more symptoms associated with the condition or prevent its
recurrence. A prophylactically effective amount of a compound means
an amount of a therapeutic agent, alone or in combination with
other agents, which provides a prophylactic benefit in the
prevention of the condition. The term "prophylactically effective
amount" can encompass an amount that improves overall prophylaxis
or enhances the prophylactic efficacy of another prophylactic
agent.
[0105] A "proliferative disease" refers to a disease that occurs
due to abnormal growth or extension by the multiplication of cells
(Walker, Cambridge Dictionary of Biology; Cambridge University
Press: Cambridge, UK, 1990). A proliferative disease may be
associated with: 1) the pathological proliferation of normally
quiescent cells; 2) the pathological migration of cells from their
normal location (e.g., metastasis of neoplastic cells); 3) the
pathological expression of proteolytic enzymes such as the matrix
metalloproteinases (e.g., collagenases, gelatinases, and
elastases); or 4) the pathological angiogenesis as in proliferative
retinopathy and tumor metastasis. Exemplary proliferative diseases
include cancers (i.e., "malignant neoplasms"), benign neoplasms,
diseases associated with angiogenesis, inflammatory diseases, and
autoimmune diseases.
[0106] The term "angiogenesis" refers to the physiological process
through which new blood vessels form from pre-existing vessels.
Angiogenesis is distinct from vasculogenesis, which is the de novo
formation of endothelial cells from mesoderm cell precursors. The
first vessels in a developing embryo form through vasculogenesis,
after which angiogenesis is responsible for most blood vessel
growth during normal or abnormal development. Angiogenesis is a
vital process in growth and development, as well as in wound
healing and in the formation of granulation tissue. However,
angiogenesis is also a fundamental step in the transition of tumors
from a benign state to a malignant one, leading to the use of
angiogenesis inhibitors in the treatment of cancer. Angiogenesis
may be chemically stimulated by angiogenic proteins, such as growth
factors (e.g., VEGF). "Pathological angiogenesis" refers to
abnormal (e.g., excessive or insufficient) angiogenesis that
amounts to and/or is associated with a disease.
[0107] The terms "neoplasm" and "tumor" are used herein
interchangeably and refer to an abnormal mass of tissue wherein the
growth of the mass surpasses and is not coordinated as in the
growth of normal tissue. A neoplasm or tumor may be "benign" or
"malignant," depending on the following characteristics: degree of
cellular differentiation (including morphology and functionality),
rate of growth, local invasion, and metastasis. A "benign neoplasm"
is generally well differentiated, has characteristically slower
growth than a malignant neoplasm, and remains localized to the site
of origin. In addition, a benign neoplasm does not have the
capacity to infiltrate, invade, or metastasize to distant sites.
Exemplary benign neoplasms include, but are not limited to, lipoma,
chondroma, adenomas, acrochordon, senile angiomas, seborrheic
keratoses, lentigos, and sebaceous hyperplasias. In some cases,
certain "benign" tumors may later give rise to malignant neoplasms,
which may result from additional genetic changes in a subpopulation
of the tumor's neoplastic cells, and these tumors are referred to
as "pre-malignant neoplasms." An exemplary pre-malignant neoplasm
is a teratoma. In contrast, a "malignant neoplasm" is generally
poorly differentiated (anaplasia) and has characteristically rapid
growth accompanied by progressive infiltration, invasion, and
destruction of the surrounding tissue. Furthermore, a malignant
neoplasm generally has the capacity to metastasize to distant
sites. The term "metastasis," "metastatic," or "metastasize" refers
to the spread or migration of cancerous cells from a primary or
original tumor to another organ or tissue and is typically
identifiable by the presence of a "secondary tumor" or "secondary
cell mass" of the tissue type of the primary or original tumor and
not of that of the organ or tissue in which the secondary
(metastatic) tumor is located. For example, a prostate cancer that
has migrated to bone is said to be metastasized prostate cancer and
includes cancerous prostate cancer cells growing in bone
tissue.
[0108] The term "cancer" refers to a class of diseases
characterized by the development of abnormal cells that proliferate
uncontrollably and have the ability to infiltrate and destroy
normal body tissues. See, e.g., Stedman's Medical Dictionary, 25th
ed.; Hensyl ed.; Williams & Wilkins: Philadelphia, 1990.
Exemplary cancers include, but are not limited to, hematological
malignancies. Additional exemplary cancers include, but are not
limited to, acoustic neuroma; adenocarcinoma; adrenal gland cancer,
anal cancer; angiosarcoma (e.g., lymphangiosarcoma,
lymphangioendotheliosarcoma, hemangiosarcoma); appendix cancer;
benign monoclonal gammopathy; biliary cancer (e.g.,
cholangiocarcinoma); bladder cancer; breast cancer (e.g.,
adenocarcinoma of the breast, papillary carcinoma of the breast,
mammary cancer, medullary carcinoma of the breast, triple negative
breast cancer (TNBC)); brain cancer (e.g., meningioma,
glioblastomas, glioma (e.g., astrocytoma, oligodendroglioma),
medulloblastoma); bronchus cancer; carcinoid tumor; cervical cancer
(e.g., cervical adenocarcinoma); choriocarcinoma; chordoma;
craniopharyngioma; colorectal cancer (e.g., colon cancer, rectal
cancer, colorectal adenocarcinoma); connective tissue cancer;
epithelial carcinoma; ependymoma; endotheliosarcoma (e.g., Kaposi's
sarcoma, multiple idiopathic hemorrhagic sarcoma); endometrial
cancer (e.g., uterine cancer, uterine sarcoma); esophageal cancer
(e.g., adenocarcinoma of the esophagus, Barrett's adenocarcinoma);
Ewing's sarcoma; ocular cancer (e.g., intraocular melanoma,
retinoblastoma); familiar hypereosinophilia; gall bladder cancer;
gastric cancer (e.g., stomach adenocarcinoma); gastrointestinal
stromal tumor (GIST); germ cell cancer; head and neck cancer (e.g.,
head and neck squamous cell carcinoma, oral cancer (e.g., oral
squamous cell carcinoma), throat cancer (e.g., laryngeal cancer,
pharyngeal cancer, nasopharyngeal cancer, oropharyngeal cancer));
heavy chain disease (e.g., alpha chain disease, gamma chain
disease, mu chain disease; hemangioblastoma; hypopharynx cancer;
inflammatory myofibroblastic tumors; immunocytic amyloidosis;
kidney cancer (e.g., nephroblastoma a.k.a. Wilms' tumor, renal cell
carcinoma); liver cancer (e.g., hepatocellular cancer (HCC),
malignant hepatoma); lung cancer (e.g., bronchogenic carcinoma,
small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC),
adenocarcinoma of the lung); leiomyosarcoma (LMS); mastocytosis
(e.g., systemic mastocytosis); muscle cancer; myelodysplastic
syndrome (MDS); mesothelioma; myeloproliferative disorder (MPD)
(e.g., polycythemia vera (PV), essential thrombocytosis (ET),
agnogenic myeloid metaplasia (AMM) a.k.a. myelofibrosis (MF),
chronic idiopathic myelofibrosis, chronic myelocytic leukemia
(CML), chronic neutrophilic leukemia (CNL), hypereosinophilic
syndrome (HES)); neuroblastoma; neurofibroma (e.g.,
neurofibromatosis (NF) type 1 or type 2, schwannomatosis);
neuroendocrine cancer (e.g., gastroenteropancreatic neuroendocrine
tumor (GEP-NET), carcinoid tumor); osteosarcoma (e.g., bone
cancer); ovarian cancer (e.g., cystadenocarcinoma, ovarian
embryonal carcinoma, ovarian adenocarcinoma); papillary
adenocarcinoma; pancreatic cancer (e.g., pancreatic
andenocarcinoma, intraductal papillary mucinous neoplasm (IPMN),
Islet cell tumors); penile cancer (e.g., Paget's disease of the
penis and scrotum); pinealoma; primitive neuroectodermal tumor
(PNT); plasma cell neoplasia; paraneoplastic syndromes;
intraepithelial neoplasms; prostate cancer (e.g., prostate
adenocarcinoma); rectal cancer; rhabdomyosarcoma; salivary gland
cancer; skin cancer (e.g., squamous cell carcinoma (SCC),
keratoacanthoma (KA), melanoma, basal cell carcinoma (BCC)); small
bowel cancer (e.g., appendix cancer); soft tissue sarcoma (e.g.,
malignant fibrous histiocytoma (MFH), liposarcoma, malignant
peripheral nerve sheath tumor (MPNST), chondrosarcoma,
fibrosarcoma, myxosarcoma); sebaceous gland carcinoma; small
intestine cancer; sweat gland carcinoma; synovioma; testicular
cancer (e.g., seminoma, testicular embryonal carcinoma); thyroid
cancer (e.g., papillary carcinoma of the thyroid, papillary thyroid
carcinoma (PTC), medullary thyroid cancer); urethral cancer;
vaginal cancer; and vulvar cancer (e.g., Paget's disease of the
vulva).
[0109] The term "hematological malignancy" refers to tumors that
affect blood, bone marrow, and/or lymph nodes. Exemplary
hematological malignancies include, but are not limited to,
leukemia, such as acute lymphoblastic leukemia (ALL) (e.g., B-cell
ALL, T-cell ALL), acute myelocytic leukemia (AML) (e.g., B-cell
AML, T-cell AML), chronic myelocytic leukemia (CML) (e.g., B-cell
CML, T-cell CML), and chronic lymphocytic leukemia (CLL) (e.g.,
B-cell CLL, T-cell CLL)); lymphoma, such as Hodgkin lymphoma (HL)
(e.g., B-cell HL, T-cell HL) and non-Hodgkin lymphoma (NHL) (e.g.,
B-cell NHL, such as diffuse large cell lymphoma (DLCL) (e.g.,
diffuse large B-cell lymphoma (DLBCL, e.g., activated B-cell (ABC)
DLBCL (ABC-DLBCL))), follicular lymphoma, chronic lymphocytic
leukemia/small lymphocytic lymphoma (CLL/SLL), mantle cell lymphoma
(MCL), marginal zone B-cell lymphoma (e.g., mucosa-associated
lymphoid tissue (MALT) lymphoma, nodal marginal zone B-cell
lymphoma, splenic marginal zone B-cell lymphoma), primary
mediastinal B-cell lymphoma, Burkitt's lymphoma, Waldenstrom's
macroglobulinemia (WM, lymphoplasmacytic lymphoma), hairy cell
leukemia (HCL), immunoblastic large cell lymphoma, precursor
B-lymphoblastic lymphoma, central nervous system (CNS) lymphoma
(e.g., primary CNS lymphoma and secondary CNS lymphoma); and T-cell
NHL, such as precursor T-lymphoblastic lymphoma/leukemia,
peripheral T-cell lymphoma (PTCL) (e.g., cutaneous T-cell lymphoma
(CTCL) (e.g., mycosis fungoides, Sezary syndrome),
angioimmunoblastic T-cell lymphoma, extranodal natural killer
T-cell lymphoma, enteropathy type T-cell lymphoma, subcutaneous
panniculitis-like T-cell lymphoma, and anaplastic large cell
lymphoma); lymphoma of an immune privileged site (e.g., cerebral
lymphoma, ocular lymphoma, lymphoma of the placenta, lymphoma of
the fetus, testicular lymphoma); a mixture of one or more
leukemia/lymphoma as described above; myelodysplasia; and multiple
myeloma (MM).
[0110] The term "inflammatory disease" refers to a disease caused
by, resulting from, or resulting in inflammation. The term
"inflammatory disease" may also refer to a dysregulated
inflammatory reaction that causes an exaggerated response by
macrophages, granulocytes, and/or T-lymphocytes leading to abnormal
tissue damage and/or cell death. An inflammatory disease can be
either an acute or chronic inflammatory condition and can result
from infections or non-infectious causes. Inflammatory diseases
include, without limitation, atherosclerosis, arteriosclerosis,
autoimmune disorders, multiple sclerosis, systemic lupus
erythematosus, polymyalgia rheumatica (PMR), gouty arthritis,
degenerative arthritis, tendonitis, bursitis, psoriasis, cystic
fibrosis, arthrosteitis, rheumatoid arthritis, inflammatory
arthritis, Sjogren's syndrome, giant cell arteritis, progressive
systemic sclerosis (scleroderma), ankylosing spondylitis,
polymyositis, dermatomyositis, pemphigus, pemphigoid, diabetes
(e.g., Type I), myasthenia gravis, Hashimoto's thyroiditis, Graves'
disease, Goodpasture's disease, mixed connective tissue disease,
sclerosing cholangitis, inflammatory bowel disease, Crohn's
disease, ulcerative colitis, pernicious anemia, inflammatory
dermatoses, usual interstitial pneumonitis (UIP), asbestosis,
silicosis, bronchiectasis, berylliosis, talcosis, pneumoconiosis,
sarcoidosis, desquamative interstitial pneumonia, lymphoid
interstitial pneumonia, giant cell interstitial pneumonia, cellular
interstitial pneumonia, extrinsic allergic alveolitis, Wegener's
granulomatosis and related forms of angiitis (temporal arteritis
and polyarteritis nodosa), inflammatory dermatoses, hepatitis,
delayed-type hypersensitivity reactions (e.g., poison ivy
dermatitis), pneumonia, respiratory tract inflammation, Adult
Respiratory Distress Syndrome (ARDS), encephalitis, immediate
hypersensitivity reactions, asthma, hay fever, allergies, acute
anaphylaxis, rheumatic fever, glomerulonephritis, pyelonephritis,
cellulitis, cystitis, chronic cholecystitis, ischemia (ischemic
injury), reperfusion injury, allograft rejection, host-versus-graft
rejection, appendicitis, arteritis, blepharitis, bronchiolitis,
bronchitis, cervicitis, cholangitis, chorioamnionitis,
conjunctivitis, dacryoadenitis, dermatomyositis, endocarditis,
endometritis, enteritis, enterocolitis, epicondylitis,
epididymitis, fasciitis, fibrositis, gastritis, gastroenteritis,
gingivitis, ileitis, iritis, laryngitis, myelitis, myocarditis,
nephritis, omphalitis, oophoritis, orchitis, osteitis, otitis,
pancreatitis, parotitis, pericarditis, pharyngitis, pleuritis,
phlebitis, pneumonitis, proctitis, prostatitis, rhinitis,
salpingitis, sinusitis, stomatitis, synovitis, testitis,
tonsillitis, urethritis, urocystitis, uveitis, vaginitis,
vasculitis, vulvitis, vulvovaginitis, angitis, chronic bronchitis,
osteomyelitis, optic neuritis, temporal arteritis, transverse
myelitis, necrotizing fasciitis, and necrotizing enterocolitis.
[0111] An "autoimmune disease" refers to a disease arising from an
inappropriate immune response of the body of a subject against
substances and tissues normally present in the body. In other
words, the immune system mistakes some part of the body as a
pathogen and attacks its own cells. This may be restricted to
certain organs (e.g., in autoimmune thyroiditis) or involve a
particular tissue in different places (e.g., Goodpasture's disease
which may affect the basement membrane in both the lung and
kidney). The treatment of autoimmune diseases is typically with
immunosuppression, e.g., medications which decrease the immune
response. Exemplary autoimmune diseases include, but are not
limited to, glomerulonephritis, Goodpasture's syndrome, necrotizing
vasculitis, lymphadenitis, peri-arteritis nodosa, systemic lupus
erythematosis, rheumatoid arthritis, psoriatic arthritis, systemic
lupus erythematosis, psoriasis, ulcerative colitis, systemic
sclerosis, dermatomyositis/polymyositis, anti-phospholipid antibody
syndrome, scleroderma, pemphigus vulgaris, ANCA-associated
vasculitis (e.g., Wegener's granulomatosis, microscopic
polyangiitis), uveitis, Sjogren's syndrome, Crohn's disease,
Reiter's syndrome, ankylosing spondylitis, Lyme disease,
Guillain-Barre syndrome, Hashimoto's thyroiditis, and
cardiomyopathy.
[0112] The term "kinase" is a class of enzyme that transfers a
phosphate group from a high energy donor molecules, such as ATP, to
a specific substrate, referred to as phosphorylation. Kinases are
part of the larger family of phosphotransferases. One of the
largest groups of kinases are protein kinases, which act on and
modify the activity of specific proteins. Kinases are used
extensively to transmit signals and control complex processes in
cells. Various other kinases act on small molecules such as lipids,
carbohydrates, amino acids, and nucleotides, either for signaling
or to prime them for metabolic pathways. Kinases are often named
after their substrates. More than 500 different protein kinases
have been identified in humans. Exemplary human protein kinases
include, but are not limited to, AAK1, ABL, ACK, ACTR2, ACTR2B,
AKT1, AKT2, AKT3, ALK, ALK1, ALK2, ALK4, ALK7, AMPKa1, AMPKa2,
ANKRD3, ANPa, ANPb, ARAF, ARAFps, ARG, AurA, AurAps1, AurAps2,
AurB, AurBps1, AurC, AXL, BARK1, BARK2, BIKE, BLK, BMPR1A,
BMPR1Aps1, BMPR1Aps2, BMPR1B, BMPR2, BMX, BRAF, BRAFps, BRK, BRSK1,
BRSK2, BTK, BUB1, BUBR1, CaMK1a, CaMK1b, CaMK1d, CaMK1g, CaMK2a,
CaMK2b, CaMK2d, CaMK2g, CaMK4, CaMKK1, CaMKK2, caMLCK, CASK, CCK4,
CCRK, CDC2, CDC7, CDK10, CDK11, CDK2, CDK3, CDK4, CDK4ps, CDK5,
CDK5ps, CDK6, CDK7, CDK7ps, CDK8, CDK8ps, CDK9, CDKL1, CDKL2,
CDKL3, CDKL4, CDKL5, CGDps, CHED, CHK1, CHK2, CHK2ps1, CHK2ps2,
CK1a, CK1a2, CK1aps1, CK1aps2, CK1aps3, CK1d, CK1e, CK1g1, CK1g2,
CK1g2ps, CK1g3, CK2a1, CK2a1-rs, CK2a2, CLIK1, CLIK1L, CLK1, CLK2,
CLK2ps, CLK3, CLK3ps, CLK4, COT, CRIK, CRK7, CSK, CTK, CYGD, CYGF,
DAPK1, DAPK2, DAPK3, DCAMKL1, DCAMKL2, DCAMKL3, DDR1, DDR2, DLK,
DMPK1, DMPK2, DRAK1, DRAK2, DYRK1A, DYRK1B, DYRK2, DYRK3, DYRK4,
EGFR, EphA1, EphA10, EphA2, EphA3, EphA4, EphA5, EphA6, EphA7,
EphA8, EphB1, EphB2, EphB3, EphB4, EphB6, Erk1, Erk2, Erk3,
Erk3ps1, Erk3ps2, Erk3ps3, Erk3ps4, Erk4, Erk5, Erk7, FAK, FER,
FERps, FES, FGFR1, FGFR2, FGFR3, FGFR4, FGR, FLT1, FLT1ps, FLT3,
FLT4, FMS, FRK, Fused, FYN, GAK, GCK, GCN2, GCN22, GPRK4, GPRK5,
GPRK6, GPRK6ps, GPRK7, GSK3A, GSK3B, Haspin, HCK, HER2/ErbB2,
HER3/ErbB3, HER4/ErbB4, HH498, HIPK1, HIPK2, HIPK3, HIPK4, HPK1,
HRI, HRIps, HSER, HUNK, ICK, IGF1R, IKKa, IKKb, IKKe, ILK, INSR,
IRAK1, IRAK2, IRAK3, IRAK4, IRE1, IRE2, IRR, ITK, JAK1, JAK2, JAK3,
JNK1, JNK2, JNK3, KDR, KHS1, KHS2, KIS, KIT, KSGCps, KSR1, KSR2,
LATS1, LATS2, LCK, LIMK1, LIMK2, LIMK2ps, LKB1, LMR1, LMR2, LMR3,
LOK, LRRK1, LRRK2, LTK, LYN, LZK, MAK, MAP2K1, MAP2K1ps, MAP2K2,
MAP2K2ps, MAP2K3, MAP2K4, MAP2K5, MAP2K6, MAP2K7, MAP3K1, MAP3K2,
MAP3K3, MAP3K4, MAP3K5, MAP3K6, MAP3K7, MAP3K8, MAPKAPK2, MAPKAPK3,
MAPKAPK5, MAPKAPKps1, MARK1, MARK2, MARK3, MARK4, MARKps01,
MARKps02, MARKps03, MARKps04, MARKps05, MARKps07, MARKps08,
MARKps09, MARKps10, MARKps11, MARKps12, MARKps13, MARKps15,
MARKps16, MARKps17, MARKps18, MARKps19, MARKps20, MARKps21,
MARKps22, MARKps23, MARKps24, MARKps25, MARKps26, MARKps27,
MARKps28, MARKps29, MARKps30, MAST1, MAST2, MAST3, MAST4, MASTL,
MELK, MER, MET, MISR2, MLK1, MLK2, MLK3, MLK4, MLKL, MNK1, MNK1ps,
MNK2, MOK, MOS, MPSK1, MPSK1ps, MRCKa, MRCKb, MRCKps, MSK1, MSK12,
MSK2, MSK22, MSSK1, MST1, MST2, MST3, MST3ps, MST4, MUSK, MYO3A,
MYO3B, MYT1, NDR1, NDR2, NEK1, NEK10, NEK11, NEK2, NEK2ps1,
NEK2ps2, NEK2ps3, NEK3, NEK4, NEK4ps, NEK5, NEK6, NEK7, NEK8, NEK9,
NIK, NIM1, NLK, NRBP1, NRBP2, NuaK1, NuaK2, Obscn, Obscn2, OSR1,
p38a, p38b, p38d, p38g, p70S6K, p70S6Kb, p70S6Kps1, p70S6Kps2,
PAK1, PAK2, PAK2ps, PAK3, PAK4, PAK5, PAK6, PASK, PBK, PCTAIRE1,
PCTAIRE2, PCTAIRE3, PDGFRa, PDGFRb, PDK1, PEK, PFTAIRE1, PFTAIRE2,
PHKg1, PHKg1ps1, PHKg1ps2, PHKg1ps3, PHKg2, PIK3R4, PIM1, PIM2,
PIM3, PINK1, PITSLRE, PKACa, PKACb, PKACg, PKCa, PKCb, PKCd, PKCe,
PKCg, PKCh, PKCi, PKCips, PKCt, PKCz, PKD1, PKD2, PKD3, PKG1, PKG2,
PKN1, PKN2, PKN3, PKR, PLK1, PLK1ps1, PLK1ps2, PLK2, PLK3, PLK4,
PRKX, PRKXps, PRKY, PRP4, PRP4ps, PRPK, PSKH1, PSKH1ps, PSKH2,
PYK2, QIK, QSK, RAF1, RAF1ps, RET, RHOK, RIPK1, RIPK2, RIPK3,
RNAseL, ROCK1, ROCK2, RON, ROR1, ROR2, ROS, RSK1, RSK12, RSK2,
RSK22, RSK3, RSK32, RSK4, RSK42, RSKL1, RSKL2, RYK, RYKps, SAKps,
SBK, SCYL1, SCYL2, SCYL2ps, SCYL3, SGK, SgK050ps, SgK069, SgK071,
SgK085, SgK110, SgK196, SGK2, SgK223, SgK269, SgK288, SGK3, SgK307,
SgK384ps, SgK396, SgK424, SgK493, SgK494, SgK495, SgK496, SIK
(e.g., SIK1, SIK2), skMLCK, SLK, Slob, smMLCK, SNRK, SPEG, SPEG2,
SRC, SRM, SRPK1, SRPK2, SRPK2ps, SSTK, STK33, STK33ps, STLK3,
STLK5, STLK6, STLK6ps1, STLK6-rs, SuRTK106, SYK, TAK1, TAO1, TAO2,
TAO3, TBCK, TBK1, TEC, TESK1, TESK2, TGFbR1, TGFbR2, TIE1, TIE2,
TLK1, TLK1ps, TLK2, TLK2ps1, TLK2ps2, TNK1, Trad, Trb1, Trb2, Trb3,
Trio, TRKA, TRKB, TRKC, TSSK1, TSSK2, TSSK3, TSSK4, TSSKps1,
TSSKps2, TTBK1, TTBK2, TTK, TTN, TXK, TYK2, TYK22, TYRO3, TYRO3ps,
ULK1, ULK2, ULK3, ULK4, VACAMKL, VRK1, VRK2, VRK3, VRK3ps, Wee1,
Wee1B, Wee1Bps, Wee1ps1, Wee1ps2, Wnk1, Wnk2, Wnk3, Wnk4, YANK1,
YANK2, YANK3, YES, YESps, YSK1, ZAK, ZAP70, ZC1/HGK, ZC2/TNIK,
ZC3/MINK, and ZC4/NRK.
[0113] The term "SRC family kinase" refers to a family of
non-receptor tyrosine protein kinases that includes nine members:
SRCA subfamily that includes c-SRC (proto-oncogene tyrosine-protein
kinase SRC), YES (proto-oncogene tyrosine-protein kinase Yes), FYN
(proto-oncogene tyrosine-protein kinase FYN), and FGR
(Gardner-Rasheed feline sarcoma viral (v-FGR) oncogene homolog);
SRCB subfamily that includes LCK (lymphocyte-specific protein
tyrosine kinase), HCK (tyrosine-protein kinase HCK, hemopoietic
cell kinase), BLK (tyrosine-protein kinase BLK), and LYN
(tyrosine-protein kinase LYN); and FRK (Fyn-related kinase).
[0114] The term "CDK" refers to a cyclin-dependent kinase. A CDK
binds a cyclin (e.g., Cyclin H), which is a regulatory protein.
CDKs phosphorylate their substrates at serines and threonines. The
consensus sequence for the phosphorylation site in the amino acid
sequence of a CDK substrate is [S/T*]PX[K/R], where S/T* is the
phosphorylated serine or threonine, P is proline, X is any amino
acid, K is lysine, and R is arginine. CDKs include CDK1, CDK2,
CDK3, CDK4, CDK5, CDK6, CDK7, CDK8, CDK9, CDK10, CDK11, CDK12,
CDK13, CDK14, CDK15, CDK16, CDK17, CDK18, CDK19. and CDK20.
[0115] CDK7, cyclin-dependent kinase 7, is a CDK, wherein the
substrate is Cyclin H, MAT1 (e.g., MNAT1), or Cyclin H and MAT1.
CDK7 is alternatively referred to as CAK1, HCAK, MO15, STK1, CDKN7,
and p39MO15. Non-limiting examples of the nucleotide and protein
sequences for human CDK7 are described in GenBank Accession Number:
NP_001790, incorporated herein by reference. The amino acid
sequence of this CDK7 is as follows:
TABLE-US-00001 (SEQ ID NO: 1)
MALDVKSRAKRYEKLDFLGEGQFATVYKARDKNTNQIVAIKKIKLGHRSE
AKDGINRTALREIKLLQELSHPNIIGLLDAFGHKSNISLVEDFMETDLEV
IIKDNSLVLTPSHIKAYMLMTLQGLEYLHQHWILHRDLKPNNLLLDENGV
LKLADFGLAKSFGSPNRAYTHQVVTRWYRAPELLFGARMYGVGVDMWAVG
CILAELLLRVPFLPGDSDLDQLTRIFETLGTPTEEQWPDMCSLPDYVTFK
SFPGIPLHHIFSAAGDDLLDLIQGLFLFNPCARITATQALKMKYFSNRPG
PTPGCQLPRPNCPVETLKEQSNPALAIKRKRTEALEQGGLPKKLIF
[0116] CDK12, cyclin-dependent kinase 12, is a CDK, wherein the
substrate is Cyclin K or flavopiridol. CDK12 is alternatively
referred to as Cdc2-related kinase, CDC2-related protein kinase 7,
Cell division cycle 2-related protein kinase 7, Cell division
protein kinase 12, CRK7, CRKR, CRKRS, cyclin-dependent kinase 12,
or KIAA0904. Non-limiting examples of the nucleotide and protein
sequences for human CDK12 are described in Uniprot Number: Q9NYV4,
which is incorporated herein by reference. The amino acid sequence
of this CDK12 is as follows:
TABLE-US-00002 (SEQ ID NO 2)
MPNSERHGGKKDGSGGASGTLQPSSGGGSSNSRERHRLVSKHKRHKSKHS
KDMGLVTPEAASLGTVIKPLVEYDDISSDSDTFSDDMAFKLDRRENDERR
GSDRSDRLHKHRHHQHRRSRDLLKAKQTEKEKSQEVSSKSGSMKDRISGS
SKRSNEETDDYGKAQVAKSSSKESRSSKLHKEKTRKERELKSGHKDRSKS
HRKRETPKSYKTVDSPKRRSRSPHRKWSDSSKQDDSPSGASYGQDYDLSP
SRSHTSSNYDSYKKSPGSTSRRQSVSPPYKEPSAYQSSTRSPSPYSKRQR
SVSPYSRRRSSSYERSGSYSGRSPSPYGRRRSSSPFLSKRSLSRSPLPSR
KSMKSRSRSPAYSRHSSSHSKKKRSSSRSRHSSISPVRLPLNSSLGAELS
RKKKERAAAAAAAKMDGKESKGSPVFLPRKENSSVEAKDSGLESKKLPRS
VKLEKSAPDTELVNVTHLNTEVKNSSDTGKVKLDENSEKHLVKDLKAQGT
RDSKPIALKEEIVTPKETETSEKETPPPLPTIASPPPPLPTTTPPPQTPP
LPPLPPIPALPQQPPLPPSQPAFSQVPASSTSTLPPSTHSKTSAVSSQAN
SQPPVQVSVKTQVSVTAAIPHLKTSTLPPLPLPPLLPGDDDMDSPKETLP
SKPVKKEKEQRTRHLLTDLPLPPELPGGDLSPPDSPEPKAITPPQQPYKK
RPKICCPRYGERRQTESDWGKRCVDKFDIIGIIGEGTYGQVYKAKDKDTG
ELVALKKVRLDNEKEGFPITAIREIKILRQLIHRSVVNMKEIVTDKQDAL
DFKKDKGAFYLVFEYMDHDLMGLLESGLVHFSEDHIKSFMKQLMEGLEYC
HKKNFLHRDIKCSNILLNNSGQIKLADEGLARLYNSEESRPYTNKVITLW
YRPPELLLGEERYTPAIDVWSCGCILGELFTKKPIFQANLELAQLELISR
LCGSPCPAVWPDVIKLPYFNTMKPKKQTRRRLREEFSFIPSAALDLLDHM
LTLDPSKRCTAEQTLQSDFLKDVELSKMAPPDLPHWQDCHELWSKKRRRQ
RQSGVVVEEPPPSKTSPKETTSGTSTEPVKNSSPAPPQPAPGKVESGAGD
AIGLADITQQLNQSELAVLLNLLQSQTDLSIPQMAQLLNIHSNPEMQQQL
EALNQSISALTEATSQQQDSETMAPEESLKEAPSAPVILPSAEQTTLEAS
STPADMQNILAVLLSQLMKTQEPAGSLEENNSDKNSGPQGPRRTPTMPQE
EAAACPPHILPPEKRPPEPPGPPPPPPPPPLVEGDLSSAPQELNPAVTAA
LLQLLSQPEAEPPGHLPHEHQALRPMEYSTRPRPNRTYGNTDGPETGESA
IDTDERNSGPALTESLVQTLVKNRTFSGSLSHLGESSSYQGTGSVQFPGD
QDLRFARVPLALHPVVGQPFLKAEGSSNSVVHAETKLQNYGELGPGTTGA
SSSGAGLHWGGPTQSSAYGKLYRGPTRVPPRGGRGRGVPY
[0117] CDK13, cyclin-dependent kinase 13, is a CDK, wherein the
relevant cyclin is cyclin K and a reference inhibitor is the
pan-CDK inhibitor flavopiridol and the c-terminal domain (CTD) of
RNA-polymerase II is a physiological substrate. CDK13 is
alternatively referred to as CHED; CDC2L; CDC2L5; or hCDK13.
Non-limiting examples of the nucleotide and protein sequences for
human CDK12 are described in GenBank Accession Number M80629, which
is incorporated herein by reference. The amino acid sequence of
this CDK13 is as follows:
TABLE-US-00003 (SEQ ID NO: 3)
MPSSSDTALGGGGGLSWAEKKLEERRKRRRFLSPQQPPLLLPLLQPQLLQ
PPPPPPPLLFLAAPGTAAAAAAAAAASSSCFSPGPPLEVKRLARGKRRAG
GRQKRRRGPRAGQEAEKRRVFSLPQPQQDGGGGASSGGGVTPLVEYEDVS
SQSEQGLLLGGASAATAATAAGGTGGSGGSPASSSGTQRRGEGSERRPRR
DRRSSSGRSKERHREHRRRDGQRGGSEASKSRSRESHSGEERAEVAKSGS
SSSSGGRRKSASATSSSSSSRKDRDSKAHRSRTKSSKEPPSAYKEPPKAY
REDKTEPKAYRRRRSLSPLGGRDDSPVSHRASQSLRSRKSPSPAGGGSSP
YSRRLPRSPSPYSRRRSPSYSRHSSYERGGDVSPSPYSSSSWRRSRSPYS
PVLRRSGKSRSRSPYSSRHSRSRSRHRLSRSRSRHSSISPSTLTLKSSLA
AELNKNKKARAAEAARAAEAAKAAEATKAAEAAAKAAKASNTSTPTKGNT
ETSASASQTNHVKDVKKIKIEHAPSPSSGGTLKNDKAKTKPPLQVTKVEN
NLIVDKATKYAVIVGKESKSAATKEESVSLKEKTKPLTPSIGAKEKEQHV
ALVTSTLPPLPLPPMLPEDKEADSLRGNISVKAVKKEVEKKLRCLLADLP
LPPELPGGDDLSKSPEEKKTATQLHSKRRPKICGPPYGETKEKDIDWGKR
CVDKFDISGIIGEGTYGQVYKARDKDTGEMVALKKVRLDNEKEGFPITAI
REIKILRQLTHQSIINMKEIVTDKEDALDFKKDKGAFYLVFEYMDHDLMG
LLESGLVHFNENHIKSFMRQLMEGLDYCHKKNFLHRDIKCSNILLNNRGQ
IKLADFGLARLYSSEESRPYTNKVITLWYRPPELLLGEERYTPAIDVWSC
GCILGELFTKKPIFQANQELAQLELISRICGSPCPAVWPDVIKLPYFNTM
KPKKQYRRKLREEFVFIPAAALDLFDYMLALDPSKRCTAEQALQCEFLRD
VEPSKMPPPDLPLWQDCHELWSKKRRRQKQMGMTDDVSTIKAPRKDLSLG
LDDSRTNTPQGVLPSSQLKSQGSSNVAPVKTGPGQHLNHSELAILLNLLQ
SKTSVNMADFVQVLNIKVNSETQQQLNKINLPAGILATGEKQTDPSTPQQ
ESSKPLGGIQPSSQTIQPKVETDAAQAAVQSAFAVLLTQLIKAQQSKQKD
VLLEERENGSGHEASLQLRPPPEPSTPVSGQDDLIQHQDMRILELTPEPD
RPRILPPDQRPPEPPEPPPVTEEDLDYRTENQHVPTTSSSLTDPHAGVKA
ALLQLLAQHQPQDDPKREGGIDYQAGDTYVSTSDYKDNFGSSSFSSAPYV
SNDGLGSSSAPPLERRSFIGNSDIQSLDNYSTASSHSGGPPQPSAFSESF
PSSVAGYGDIYLNAGPMLFSGDKDHRFEYSHGPIAVLANSSDPSTGPEST
HPLPAKMHNYNYGGNLQENPSGPSLMHGQTWTSPAQGPGYSQGYRGHIST STGRGRGRGLPY
DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS OF THE INVENTION
[0118] Cyclin dependent kinases (CDKs) are key regulators of the
cell cycle. Their successive activation and inactivation drives the
cycle forward. The activity of CDKs is regulated by multiple
mechanisms such as positive and negative phosphorylation, binding
of regulatory proteins like cyclins, and CDK inhibitors. CDK7 plays
a critical role in the regulation of RNA polymerase II-mediated
transcription of protein-encoding genes. Disruption of CDK7, CDK12,
and/or CDK13 signaling causes defects in transcription. However, a
complete understanding of how these disruptions affect global
transcription is lacking. Furthermore, the absence of selective
inhibitors of CDK7, CDK12, and CDK13 has hindered investigation of
the transcriptional and functional consequences of acute and
long-term inhibition of these kinases under normal and pathological
conditions. The present invention provides selective CDK7, CDK12,
and/or CDK13 inhibitors, which covalently modify a cysteine residue
located outside of the canonical kinase domain (i.e., Cys312 of
CDK7, Cys1039 of CDK12, and Cys1017 of CDK13). Selective covalent
inhibitors of these kinases may be useful in the treatment of
various proliferative diseases including cancer.
[0119] The present invention provides compounds, which inhibit the
activity of a kinase, for the prevention and/or treatment of a
subject with a proliferative disease. In certain embodiments, the
inventive compounds inhibit the activity of a cyclin-dependent
kinase (CDK). In certain embodiments, the inventive compounds
inhibit the activity of a cyclin-dependent kinase 7 (CDK7). In
certain embodiments, the inventive compounds inhibit the activity
of a cyclin-dependent kinase 12 (CDK12). In certain embodiments,
the inventive compounds inhibit the activity of a cyclin-dependent
kinase 13 (CDK13). The present invention also provides methods of
using the compounds described herein, e.g., as biological probes to
study the inhibition of the activity of a kinase (e.g., CDK (e.g.
CDK7, CDK12, and/or CDK13)), and as therapeutics, e.g., in the
prevention and/or treatment of diseases associated with the
overexpression and/or aberrant activity of a kinase (e.g., CDK
(e.g. CDK7, CDK12, and/or CDK13)). In certain embodiments, the
diseases are proliferative diseases. The proliferative diseases
that may be treated and/or prevented include, but are not limited
to, cancers (e.g., breast cancer, leukemia, melanoma, multiple
myeloma), benign neoplasms, diseases associated with angiogenesis,
inflammatory diseases, autoinflammatory diseases, and autoimmune
diseases. Also provided by the present disclosure are
pharmaceutical compositions, kits, methods, and uses including a
compound of Formulae (I'), (II'), (I), and (II) as described
herein.
Compounds
[0120] Aspects of the present disclosure relate to the compounds
described herein. The compounds described herein may be useful in
treating and/or preventing proliferative diseases in a subject,
inhibiting the activity of a protein kinase (e.g., CDK) in a
subject or biological sample, and inducing apoptosis of a cell. In
certain embodiments, a compound described herein is a compound of
any one of Formulae (I'), (II'), (I), (II), or a pharmaceutically
acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer,
stereoisomer, isotopically labeled derivative, or prodrug thereof.
In certain embodiments, a compound described herein is a compound
of Formula (I'), or a pharmaceutically acceptable salt thereof. In
certain embodiments, a compound described herein is a compound of
Formula (I), or a pharmaceutically acceptable salt thereof. In
certain embodiments, a compound described herein is a compound of
Formula (II'), or a pharmaceutically acceptable salt thereof. In
certain embodiments, a compound described herein is a compound of
Formula (II), or a pharmaceutically acceptable salt thereof.
[0121] In certain embodiments, a compound described herein is of
Formula (I'):
##STR00010##
or a pharmaceutically acceptable salt thereof, wherein:
[0122] R.sup.1 is hydrogen, halogen, or optionally substituted
alkyl;
[0123] M is O, S, or NR.sup.M;
[0124] R.sup.M is hydrogen, optionally substituted alkyl,
optionally substituted alkenyl, optionally substituted alkynyl,
optionally substituted carbocyclyl, optionally substituted
heterocyclyl, optionally substituted aryl, optionally substituted
heteroaryl, optionally substituted acyl, or a nitrogen protecting
group;
[0125] Ring A is optionally substituted monocyclic carbocyclyl,
optionally substituted monocyclic heterocyclyl, optionally
substituted phenyl, or optionally substituted monocyclic
heteroaryl;
[0126] Ring B is optionally substituted monocyclic carbocyclyl,
optionally substituted monocyclic heterocyclyl, optionally
substituted phenyl, or optionally substituted monocyclic
heteroaryl;
[0127] Ring C is optionally substituted monocyclic carbocyclyl,
optionally substituted monocyclic heterocyclyl, optionally
substituted monocyclic or bicyclic aryl, or optionally substituted
monocyclic or bicyclic heteroaryl;
[0128] each instance of R.sup.A, R.sup.B, and R.sup.C is
independently hydrogen, halogen, optionally substituted alkyl,
optionally substituted alkenyl, optionally substituted alkynyl,
optionally substituted carbocyclyl, optionally substituted
heterocyclyl, optionally substituted aryl, optionally substituted
heteroaryl, --OR.sup.a, --N(R.sup.a).sub.2, --SR.sup.a, --CN,
--SCN, --NO.sub.2, --N.sub.3, or optionally substituted acyl;
[0129] each instance of R.sup.a is independently hydrogen,
optionally substituted alkyl, optionally substituted alkenyl,
optionally substituted alkynyl, optionally substituted carbocyclyl,
optionally substituted heterocyclyl, optionally substituted aryl,
optionally substituted heteroaryl, optionally substituted acyl, a
nitrogen protecting group when attached to a nitrogen atom, an
oxygen protecting group when attached to an oxygen atom, or a
sulfur protecting group when attached to a sulfur atom, or two
instances of R.sup.a are joined to form a substituted or
unsubstituted, heterocyclic ring, or substituted or unsubstituted,
heteroaryl ring;
[0130] each of a1, b1, and c1 is independently 0, 1, 2, 3, 4, 5, or
6, as valency permits;
[0131] L.sup.1 is --CH.sub.2--, .sup.lc--S(.dbd.O).sub.2--.sup.lb,
--O--, --S--, --NR.sup.L1--, --C(.dbd.O)--,
.sup.lc--NR.sup.L1C(.dbd.O)--.sup.lb,
.sup.lc--C(.dbd.O)NR.sup.L1--.sup.lb, .sup.lc--OC(.dbd.O)--.sup.lb,
or .sup.lc--C(.dbd.O)O--.sup.lb; wherein .sup.lb indicates the
point of attachment is to Ring B; and .sup.lc indicates the point
of attachment is to Ring C;
[0132] L.sup.2 is --O--, --S--, --NR.sup.L2--,
.sup.lb--NR.sup.L2C(.dbd.O)--.sup.lm,
.sup.lb--C(.dbd.O)NR.sup.L2--.sup.lm; wherein .sup.lb indicates the
point of attachment is to Ring B; and .sup.lm indicates the point
of attachment is to the heteroaryl ring with M;
[0133] X is .sup.xm--CH.sub.2CH.sub.2--.sup.xa,
.sup.xm--CH.dbd.CH--.sup.xa, .sup.xm--CH.sub.2--NR.sup.LXxa,
.sup.xm--CH.sub.2--O--CH.sub.2--.sup.xa,
.sup.xm--CH.sub.2--NR.sup.LX--CH.sub.2--.sup.xa, --O--, --S--,
--NR.sup.LX--, .sup.xm--O--CH.sub.2--.sup.xa,
.sup.xm--S--CH.sub.2--.sup.xa,
.sup.xm--S--C(.dbd.O)CH.sub.2--.sup.xa, or
.sup.xm--NR.sup.LX--CH.sub.2--.sup.xa; wherein .sup.xa indicates
the point of attachment is to Ring A; and .sup.xm indicates the
point of attachment is to the heteroaryl ring with M;
[0134] each of R.sup.L1, R.sup.L2, and R.sup.LX is independently
hydrogen, optionally substituted C.sub.1-6 alkyl, or a nitrogen
protecting group;
[0135] R.sup.2 is any of Formulae (i-1)-(i-41):
##STR00011## ##STR00012## ##STR00013## ##STR00014##
##STR00015##
wherein: [0136] L.sup.3 is a bond or an optionally substituted
C.sub.14 hydrocarbon chain, optionally wherein one or more carbon
units of the hydrocarbon chain are independently replaced with
--C.dbd.O--, --O--, --S--, --NR.sup.L3a--, --NR.sup.L3aC(.dbd.O)--,
--C(.dbd.O)NR.sup.L3a--, --SC(.dbd.O)--, --C(.dbd.O)S--,
--OC(.dbd.O)--, --C(.dbd.O)O--, --NR.sup.L3aC(.dbd.S)--,
--C(.dbd.S)NR.sup.L3a--, trans-CR.sup.L3b.dbd.CR.sup.L3b--,
cis-CR.sup.L3b.dbd.CR.sup.L3b--, --C.dbd.C--, --S(.dbd.O)--,
--S(.dbd.O)O--, --OS(.dbd.O)--, --S(.dbd.O)NR.sup.L3a--,
--NR.sup.L3aS(.dbd.O)--, --S(.dbd.O).sub.2--, --S(.dbd.O).sub.2O--,
--OS(.dbd.O).sub.2--, --S(.dbd.O).sub.2NR.sup.L3a--, or
--NR.sup.L3aS(.dbd.O).sub.2--, wherein R.sup.L3a is hydrogen,
optionally substituted C.sub.1-6 alkyl, or a nitrogen protecting
group, and wherein each occurrence of R.sup.L3b is independently
hydrogen, halogen, optionally substituted alkyl, optionally
substituted alkenyl, optionally substituted alkynyl, optionally
substituted carbocyclyl, optionally substituted heterocyclyl,
optionally substituted aryl, or optionally substituted heteroaryl,
or two R.sup.L3b groups are joined to form an optionally
substituted carbocyclic or optionally substituted heterocyclic
ring; [0137] L.sup.4 is a bond or an optionally substituted,
branched or unbranched C.sub.1-6 hydrocarbon chain; [0138] each of
R.sup.E1, R.sup.E2, and R.sup.E3 is independently hydrogen,
halogen, optionally substituted alkyl, optionally substituted
alkenyl, optionally substituted alkynyl, optionally substituted
carbocyclyl, optionally substituted heterocyclyl, optionally
substituted aryl, optionally substituted heteroaryl, --CN,
--CH.sub.2OR.sup.EE, --CH.sub.2N(R.sup.EE).sub.2,
--CH.sub.2SR.sup.EE, --OR.sup.EE, --N(R.sup.EE).sub.2,
--Si(R.sup.EE).sub.3, and --SR.sup.EE, wherein each occurrence of
R.sup.EE is independently hydrogen, optionally substituted alkyl,
optionally substituted alkoxy, optionally substituted alkenyl,
optionally substituted alkynyl, optionally substituted carbocyclyl,
optionally substituted heterocyclyl, optionally substituted aryl,
or optionally substituted heteroaryl, or two R.sup.EE groups are
joined to form an optionally substituted heterocyclic ring; [0139]
or R.sup.E1 and R.sup.E3, or R.sup.E2 and R.sup.E3, or R.sup.E1 and
R.sup.E2 are joined to form an optionally substituted carbocyclic
or optionally substituted heterocyclic ring; [0140] R.sup.E4 is a
leaving group; [0141] R.sup.E5 is halogen; [0142] R.sup.E6 is
hydrogen, optionally substituted C.sub.1-6 alkyl, or a nitrogen
protecting group; [0143] each instance of Y is independently O, S,
or NR.sup.E7, wherein R.sup.E7 is hydrogen, optionally substituted
C.sub.1-6 alkyl, or a nitrogen protecting group; [0144] a is 1 or
2; and [0145] each instance of z is independently 0, 1, 2, 3, 4, 5,
or 6, as valency permits.
[0146] In certain embodiments, a compound described herein is of
Formula (I):
##STR00016##
or a pharmaceutically acceptable salt thereof, wherein:
[0147] R.sup.1 is hydrogen, halogen, or optionally substituted
alkyl;
[0148] M is O, S, or NR.sup.M;
[0149] R.sup.M is hydrogen, optionally substituted alkyl,
optionally substituted alkenyl, optionally substituted alkynyl,
optionally substituted carbocyclyl, optionally substituted
heterocyclyl, optionally substituted aryl, optionally substituted
heteroaryl, optionally substituted acyl, or a nitrogen protecting
group;
[0150] Ring A is optionally substituted monocyclic carbocyclyl,
optionally substituted monocyclic heterocyclyl, optionally
substituted phenyl, or optionally substituted monocyclic
heteroaryl;
[0151] Ring B is optionally substituted monocyclic carbocyclyl,
optionally substituted monocyclic heterocyclyl, optionally
substituted phenyl, or optionally substituted monocyclic
heteroaryl;
[0152] Ring C is optionally substituted monocyclic carbocyclyl,
optionally substituted monocyclic heterocyclyl, optionally
substituted phenyl, or optionally substituted monocyclic
heteroaryl;
[0153] each instance of R.sup.A, R.sup.B, and R.sup.C is
independently hydrogen, halogen, optionally substituted alkyl,
optionally substituted alkenyl, optionally substituted alkynyl,
optionally substituted carbocyclyl, optionally substituted
heterocyclyl, optionally substituted aryl, optionally substituted
heteroaryl, --OR.sup.a, --N(R.sup.a).sub.2, --SR.sup.a, --CN,
--SCN, --NO.sub.2, --N.sub.3, or optionally substituted acyl;
[0154] each instance of R.sup.a is independently hydrogen,
optionally substituted alkyl, optionally substituted alkenyl,
optionally substituted alkynyl, optionally substituted carbocyclyl,
optionally substituted heterocyclyl, optionally substituted aryl,
optionally substituted heteroaryl, optionally substituted acyl, a
nitrogen protecting group when attached to a nitrogen atom, an
oxygen protecting group when attached to an oxygen atom, or a
sulfur protecting group when attached to a sulfur atom, or two
instances of R.sup.a are joined to form a substituted or
unsubstituted, heterocyclic ring, or substituted or unsubstituted,
heteroaryl ring;
[0155] each of a1, b1, and c1 is independently 0, 1, 2, 3, 4, 5, or
6, as valency permits;
[0156] L.sup.1 is --O--, --S--, --NR.sup.L1--, --C(.dbd.O)--,
.sup.lc--NR.sup.L1C(.dbd.O)--.sup.lb,
.sup.lc--C(.dbd.O)NR.sup.L1--.sup.lb, .sup.lc--OC(.dbd.O)--.sup.lb,
or .sup.lc--C(.dbd.O)O--.sup.lb; wherein .sup.lb indicates the
point of attachment is to Ring B; and .sup.lc indicates the point
of attachment is to Ring C;
[0157] L.sup.2 is --O--, --S--, --NR.sup.L2--,
.sup.lb--NR.sup.L2C(.dbd.O)--.sup.lm,
.sup.lb--C(.dbd.O)NR.sup.L2--.sup.lm; wherein .sup.lb indicates the
point of attachment is to Ring B; and .sup.lm indicates the point
of attachment is to the heteroaryl ring with M;
[0158] X is --O--, --S--, --NR.sup.LX--,
.sup.xm--O--CH.sub.2--.sup.xa, .sup.xm--S--CH.sub.2--.sup.xa, or
.sup.xm--NR.sup.LX--CH.sub.2--.sup.xa; wherein .sup.xa indicates
the point of attachment is to Ring A; and .sup.xm indicates the
point of attachment is to the heteroaryl ring with M;
[0159] each of R.sup.L1, R.sup.L2, and R.sup.LX is independently
hydrogen, optionally substituted C.sub.1-6 alkyl, or a nitrogen
protecting group;
[0160] R.sup.2 is any of Formulae (i-1)-(i-46):
##STR00017## ##STR00018## ##STR00019## ##STR00020##
##STR00021##
[0161] L.sup.3 is a bond or an optionally substituted C.sub.1-4
hydrocarbon chain, optionally wherein one or more carbon units of
the hydrocarbon chain are independently replaced with --C.dbd.O--,
--O--, --S--, --NR.sup.L3a--, --NR.sup.L3aC(.dbd.O)--,
--C(.dbd.O)NR.sup.L3a--, --SC(.dbd.O)--, --C(.dbd.O)S--,
--OC(.dbd.O)--, --C(.dbd.O)O--, --NR.sup.L3aC(.dbd.S)--,
--C(.dbd.S)NR.sup.L3a--, trans-CR.sup.L3b.dbd.CR.sup.L3b--,
cis-CR.sup.L3b.dbd.CR.sup.L3b, --C.ident.C--, --S(.dbd.O)--,
--S(.dbd.O)O--, --OS(.dbd.O)--, --S(.dbd.O)NR.sup.L3a--,
--NR.sup.L3aS(.dbd.O)--, --S(.dbd.O).sub.2--, --S(.dbd.O).sub.2O--,
--OS(.dbd.O).sub.2--, --S(.dbd.O).sub.2NR.sup.L3a, or
--NR.sup.L3aS(.dbd.O).sub.2--, wherein R.sup.L3a is hydrogen,
substituted or unsubstituted C.sub.1-6 alkyl, or a nitrogen
protecting group, and wherein each occurrence of R.sup.L3b is
independently hydrogen, halogen, optionally substituted alkyl,
optionally substituted alkenyl, optionally substituted alkynyl,
optionally substituted carbocyclyl, optionally substituted
heterocyclyl, optionally substituted aryl, or optionally
substituted heteroaryl, or two R.sup.L3b groups are joined to form
an optionally substituted carbocyclic or optionally substituted
heterocyclic ring;
[0162] L.sup.4 is a bond or an optionally substituted, branched or
unbranched C.sub.1 hydrocarbon chain;
[0163] each of R.sup.E1, R.sup.E2, and R.sup.E3 is independently
hydrogen, halogen, optionally substituted alkyl, optionally
substituted alkenyl, optionally substituted alkynyl, optionally
substituted carbocyclyl, optionally substituted heterocyclyl,
optionally substituted aryl, optionally substituted heteroaryl,
--CN, --CH.sub.2OR.sup.EE, --CH.sub.2N(R.sup.EE).sub.2,
--CH.sub.2SR.sup.EE, --OR.sup.EE, --N(R.sup.EE).sub.2,
--Si(R.sup.EE).sub.3, and --SR.sup.EE, wherein each occurrence of
R.sup.EE is independently hydrogen, optionally substituted alkyl,
optionally substituted alkoxy, optionally substituted alkenyl,
optionally substituted alkynyl, optionally substituted carbocyclyl,
optionally substituted heterocyclyl, optionally substituted aryl,
or optionally substituted heteroaryl, or two R.sup.EE groups are
joined to form an optionally substituted heterocyclic ring; or
R.sup.E1 and R.sup.E3, or R.sup.E2 and R.sup.E3, or R.sup.E1 and
R.sup.E2 are joined to form an optionally substituted carbocyclic
or optionally substituted heterocyclic ring;
[0164] R.sup.E4 is a leaving group;
[0165] R.sup.E5 is halogen;
[0166] R.sup.E6 is hydrogen, substituted or unsubstituted C.sub.1-6
alkyl, or a nitrogen protecting group;
[0167] each instance of Y is independently O, S, or NR.sup.E7,
wherein R.sup.E7 is hydrogen, substituted or unsubstituted
C.sub.1-6 alkyl, or a nitrogen protecting group;
[0168] a is 1 or 2; and
[0169] each instance of z is independently 0, 1, 2, 3, 4, 5, or 6,
as valency permits.
[0170] In certain embodiments, a compound described herein is of
Formula (II'):
##STR00022##
[0171] R.sup.1 is hydrogen, halogen, or optionally substituted
alkyl;
[0172] M is O, S, or NR.sup.M;
[0173] R.sup.M is hydrogen, optionally substituted alkyl,
optionally substituted alkenyl, optionally substituted alkynyl,
optionally substituted carbocyclyl, optionally substituted
heterocyclyl, optionally substituted aryl, optionally substituted
heteroaryl, optionally substituted acyl, or a nitrogen protecting
group;
[0174] Ring A is optionally substituted monocyclic carbocyclyl,
optionally substituted monocyclic heterocyclyl, optionally
substituted phenyl, or optionally substituted monocyclic
heteroaryl;
[0175] Ring B is optionally substituted monocyclic carbocyclyl,
optionally substituted monocyclic heterocyclyl, optionally
substituted phenyl, or optionally substituted monocyclic
heteroaryl;
[0176] each instance of R.sup.A and R.sup.B is independently
hydrogen, halogen, optionally substituted alkyl, optionally
substituted alkenyl, optionally substituted alkynyl, optionally
substituted carbocyclyl, optionally substituted heterocyclyl,
optionally substituted aryl, optionally substituted heteroaryl,
--OR.sup.a, --N(R.sup.a).sub.2, --SR.sup.a, --CN, --SCN,
--C(.dbd.NR.sup.a)R.sup.a, --C(.dbd.NR.sup.a)OR.sup.a,
--C(.dbd.NR.sup.a)N(R.sup.a).sub.2, --C(.dbd.O)R.sup.a,
--C(.dbd.O)OR.sup.a, --C(.dbd.O)N(R.sup.a).sub.2, --NO.sub.2,
--NR.sup.aC(.dbd.O)R.sup.a, --NR.sup.aC(.dbd.O)OR.sup.a,
--NR.sup.aC(.dbd.O)N(R.sup.a).sub.2, --OC(.dbd.O)R.sup.a,
--OC(.dbd.O)OR.sup.a, or --OC(.dbd.O)N(R.sup.a).sub.2;
[0177] each instance of R.sup.a is independently hydrogen,
optionally substituted acyl, optionally substituted alkyl,
optionally substituted alkenyl, optionally substituted alkynyl,
optionally substituted carbocyclyl, optionally substituted
heterocyclyl, optionally substituted aryl, optionally substituted
heteroaryl, a nitrogen protecting group when attached to a nitrogen
atom, an oxygen protecting group when attached to an oxygen atom,
or a sulfur protecting group when attached to a sulfur atom, or two
instances of R.sup.a are joined to form a substituted or
unsubstituted, heterocyclic ring, or substituted or unsubstituted,
heteroaryl ring;
[0178] each of a1 and b1 is independently 0, 1, 2, 3, 4, 5, or 6,
as valency permits;
[0179] L.sup.2 is --O--, --S--, --NR.sup.L2--,
.sup.lb--NR.sup.L2C(.dbd.O)--.sup.lm,
.sup.lb--C(.dbd.O)NR.sup.L2--.sup.lm; wherein .sup.lb indicates the
point of attachment is to Ring B; and .sup.lm indicates the point
of attachment is to the heteroaryl ring with M;
[0180] X is a bond, --O--, --S--, --NR.sup.LX--,
.sup.xm--O--CH.sub.2--.sup.xa, .sup.xm--S--CH.sub.2--.sup.xa, or
.sup.xm--NR.sup.LX--CH.sub.2--.sup.xa; wherein .sup.xa indicates
the point of attachment is to Ring A; and .sup.xm indicates the
point of attachment is to the heteroaryl ring with M;
[0181] each of R.sup.L2 and R.sup.LX is independently hydrogen,
optionally substituted C.sub.1-6 alkyl, or a nitrogen protecting
group;
[0182] R.sup.2 is any of Formulae (i-1)-(i-41):
##STR00023## ##STR00024## ##STR00025## ##STR00026##
##STR00027##
wherein: [0183] L.sup.3 is a bond or an optionally substituted
C.sub.1-4 hydrocarbon chain, optionally wherein one or more carbon
units of the hydrocarbon chain are independently replaced with
--C.dbd.O--, --O--, --S--, --NR.sup.L3a--, --NR.sup.L3aC(.dbd.O)--,
--C(.dbd.O)NR.sup.L3a--, --SC(.dbd.O)--, --C(.dbd.O)S--,
--OC(.dbd.O)--, --C(.dbd.O)O--, --NR.sup.L3aC(.dbd.S)--,
--C(.dbd.S)NR.sup.L3a--, trans-CR.sup.L3b.dbd.CR.sup.L3b--,
cis-CR.sup.L3bCR.sup.L3b--, --C.dbd.C--, --S(.dbd.O)--,
--S(.dbd.O)O--, --OS(.dbd.O)--, --S(.dbd.O)NR.sup.L3a,
--NR.sup.L3aS(.dbd.O)--, --S(.dbd.O).sub.2--, --S(.dbd.O).sub.2O--,
--OS(.dbd.O).sub.2--, --S(.dbd.O).sub.2NR.sup.L3a--, or
--NR.sup.L3aS(.dbd.O).sub.2--, wherein R.sup.L3a is hydrogen,
optionally substituted C.sub.1-6 alkyl, or a nitrogen protecting
group, and wherein each occurrence of R.sup.L3b is independently
hydrogen, halogen, optionally substituted alkyl, optionally
substituted alkenyl, optionally substituted alkynyl, optionally
substituted carbocyclyl, optionally substituted heterocyclyl,
optionally substituted aryl, or optionally substituted heteroaryl,
or two R.sup.L3b groups are joined to form an optionally
substituted carbocyclic or optionally substituted heterocyclic
ring; [0184] L.sup.4 is a bond or an optionally substituted,
branched or unbranched C.sub.1-6 hydrocarbon chain; [0185] each of
R.sup.E1, R.sup.E2, and R.sup.E3 is independently hydrogen,
halogen, optionally substituted alkyl, optionally substituted
alkenyl, optionally substituted alkynyl, optionally substituted
carbocyclyl, optionally substituted heterocyclyl, optionally
substituted aryl, optionally substituted heteroaryl, --CN,
--CH.sub.2OR.sup.EE, --CH.sub.2N(R.sup.EE).sub.2,
--CH.sub.2SR.sup.EE, --OR.sup.EE, --N(R.sup.EE).sub.2,
--Si(R.sup.EE).sub.3, and --SR.sup.EE, wherein each occurrence of
R.sup.EE is independently hydrogen, optionally substituted alkyl,
optionally substituted alkoxy, optionally substituted alkenyl,
optionally substituted alkynyl, optionally substituted carbocyclyl,
optionally substituted heterocyclyl, optionally substituted aryl,
or optionally substituted heteroaryl, or two R.sup.EE groups are
joined to form an optionally substituted heterocyclic ring; [0186]
or R.sup.E1 and R.sup.E3, or R.sup.E2 and R.sup.E3, or R.sup.E1 and
R.sup.E2 are joined to form an optionally substituted carbocyclic
or optionally substituted heterocyclic ring; [0187] R.sup.E4 is a
leaving group; [0188] R.sup.E5 is halogen; [0189] R.sup.E6 is
hydrogen, optionally substituted C.sub.14 alkyl, or a nitrogen
protecting group; each instance of Y is independently O, S, or
NR.sup.E7, wherein R.sup.E7 is hydrogen, optionally substituted
C.sub.1-6 alkyl, or a nitrogen protecting group; [0190] a is 1 or
2; and [0191] each instance of z is independently 0, 1, 2, 3, 4, 5,
or 6, as valency permits.
[0192] In certain embodiments, a compound described herein is of
Formula (I):
##STR00028##
or a pharmaceutically acceptable salt thereof, wherein:
[0193] R.sup.1 is hydrogen, halogen, or optionally substituted
alkyl;
[0194] M is O, S, or NR.sup.M;
[0195] R.sup.M is hydrogen, optionally substituted alkyl,
optionally substituted alkenyl, optionally substituted alkynyl,
optionally substituted carbocyclyl, optionally substituted
heterocyclyl, optionally substituted aryl, optionally substituted
heteroaryl, optionally substituted acyl, or a nitrogen protecting
group;
[0196] Ring A is optionally substituted monocyclic carbocyclyl,
optionally substituted monocyclic heterocyclyl, optionally
substituted phenyl, or optionally substituted monocyclic
heteroaryl;
[0197] Ring B is optionally substituted monocyclic carbocyclyl,
optionally substituted monocyclic heterocyclyl, optionally
substituted phenyl, or optionally substituted monocyclic
heteroaryl;
[0198] each instance of R.sup.A and R.sup.B is independently
hydrogen, halogen, optionally substituted alkyl, optionally
substituted alkenyl, optionally substituted alkynyl, optionally
substituted carbocyclyl, optionally substituted heterocyclyl,
optionally substituted aryl, optionally substituted heteroaryl,
--OR.sup.a, --N(R.sup.a).sub.2, --SR.sup.a, --CN, --SCN,
--C(.dbd.NR.sup.a)R.sup.a, --C(.dbd.NR.sup.a)OR.sup.a,
--C(.dbd.NR.sup.a)N(R.sup.a).sub.2, --C(.dbd.O)R.sup.a,
--C(.dbd.O)OR.sup.a, --C(.dbd.O)N(R.sup.a).sub.2, --NO.sub.2,
--NR.sup.aC(.dbd.O)R.sup.a, --NR.sup.aC(.dbd.O)OR.sup.a,
--NR.sup.aC(.dbd.O)N(R.sup.a).sub.2, --OC(.dbd.O)R.sup.a,
--OC(.dbd.O)OR.sup.a, or --OC(.dbd.O)N(R.sup.a).sub.2;
[0199] each instance of R.sup.a is independently hydrogen,
optionally substituted acyl, optionally substituted alkyl,
optionally substituted alkenyl, optionally substituted alkynyl,
optionally substituted carbocyclyl, optionally substituted
heterocyclyl, optionally substituted aryl, optionally substituted
heteroaryl, a nitrogen protecting group when attached to a nitrogen
atom, an oxygen protecting group when attached to an oxygen atom,
or a sulfur protecting group when attached to a sulfur atom, or two
instances of R.sup.a are joined to form a substituted or
unsubstituted, heterocyclic ring, or substituted or unsubstituted,
heteroaryl ring;
[0200] each of a1 and b1 is independently 0, 1, 2, 3, 4, 5, or 6,
as valency permits;
[0201] L.sup.2 is --O--, --S--, --NR.sup.L2--,
.sup.lb--NR.sup.L2C(.dbd.O)--.sup.lm,
.sup.lb--C(.dbd.O)NR.sup.L2--.sup.lm; wherein .sup.lb indicates the
point of attachment is to Ring B; and .sup.lm indicates the point
of attachment is to the heteroaryl ring with M;
[0202] X is --O--, --S--, --NR.sup.LX--,
.sup.xm--O--CH.sub.2--.sup.xa, .sup.xm--S--CH.sub.2--.sup.xa, or
.sup.xm--NR.sup.LX--CH.sub.2--.sup.xa; wherein .sup.xa indicates
the point of attachment is to Ring A; and .sup.xm indicates the
point of attachment is to the heteroaryl ring with M;
[0203] each of R.sup.L2 and R.sup.LX is independently hydrogen,
optionally substituted C.sub.1-6 alkyl, or a nitrogen protecting
group;
[0204] R.sup.2 is any of Formulae (i-1)-(i-46):
##STR00029## ##STR00030## ##STR00031## ##STR00032##
##STR00033##
wherein:
[0205] L.sup.3 is a bond or an optionally substituted C.sub.1-4
hydrocarbon chain, optionally wherein one or more carbon units of
the hydrocarbon chain are independently replaced with --C.dbd.O--,
--O--, --S--, --NR.sup.L3a--, --NR.sup.L3aC(.dbd.O)--,
--C(.dbd.O)NR.sup.L3a--, --SC(.dbd.O)--, --C(.dbd.O)S--,
--OC(.dbd.O)--, --C(.dbd.O)O--, --NR.sup.L3aC(.dbd.S)--,
--C(.dbd.S)NR.sup.L3a, trans-CR.sup.L3b.dbd.CR.sup.L3b--,
cis-CR.sup.L3b.dbd.CR.sup.L3b--, --C.ident.C--, --S(.dbd.O)--,
--S(.dbd.O)O--, --OS(.dbd.O)--, --S(.dbd.O)NR.sup.L3a--,
--NR.sup.L3aS(.dbd.O)--, --S(.dbd.O).sub.2--, --S(.dbd.O).sub.2O--,
--OS(.dbd.O).sub.2--, --S(.dbd.O).sub.2NR.sup.L3a--, or
--NR.sup.L3aS(.dbd.O).sub.2--, wherein R.sup.L3a is hydrogen,
substituted or unsubstituted C.sub.1-6 alkyl, or a nitrogen
protecting group, and wherein each occurrence of R.sup.L3b is
independently hydrogen, halogen, optionally substituted alkyl,
optionally substituted alkenyl, optionally substituted alkynyl,
optionally substituted carbocyclyl, optionally substituted
heterocyclyl, optionally substituted aryl, or optionally
substituted heteroaryl, or two R.sup.L3b groups are joined to form
an optionally substituted carbocyclic or optionally substituted
heterocyclic ring;
[0206] L.sup.4 is a bond or an optionally substituted, branched or
unbranched C.sub.1-6 hydrocarbon chain;
[0207] each of R.sup.E1, R.sup.E2, and R.sup.E3 is independently
hydrogen, halogen, optionally substituted alkyl, optionally
substituted alkenyl, optionally substituted alkynyl, optionally
substituted carbocyclyl, optionally substituted heterocyclyl,
optionally substituted aryl, optionally substituted heteroaryl,
--CN, --CH.sub.2OR.sup.EE, --CH.sub.2N(R.sup.EE).sub.2,
--CH.sub.2SR.sup.EE, --OR.sup.EE, --N(R.sup.EE).sub.2,
--Si(R.sup.EE).sub.3, and --SR.sup.EE, wherein each occurrence of
R.sup.EE is independently hydrogen, optionally substituted alkyl,
optionally substituted alkoxy, optionally substituted alkenyl,
optionally substituted alkynyl, optionally substituted carbocyclyl,
optionally substituted heterocyclyl, optionally substituted aryl,
or optionally substituted heteroaryl, or two R.sup.EE groups are
joined to form an optionally substituted heterocyclic ring;
[0208] or R.sup.E1 and R.sup.E3, or R.sup.E2 and R.sup.E3, or
R.sup.E1 and R.sup.E2 are joined to form an optionally substituted
carbocyclic or optionally substituted heterocyclic ring;
[0209] R.sup.E4 is a leaving group;
[0210] R.sup.E5 is halogen;
[0211] R.sup.E6 is hydrogen, substituted or unsubstituted C.sub.1-6
alkyl, or a nitrogen protecting group;
[0212] each instance of Y is independently O, S, or NR.sup.E7,
wherein R.sup.E7 is hydrogen, substituted or unsubstituted
C.sub.1-6 alkyl, or a nitrogen protecting group;
[0213] a is 1 or 2; and
[0214] each instance of z is independently 0, 1, 2, 3, 4, 5, or 6,
as valency permits.
[0215] As generally defined herein in Formulae (I'), (II'), (I),
and (II), R.sup.1 is hydrogen, halogen, or optionally substituted
alkyl. As generally defined herein in Formulae (I)-(II), R.sup.1 is
hydrogen, halogen, or optionally substituted alkyl. In certain
embodiments, R.sup.1 is hydrogen. In certain embodiments, R.sup.1
is halogen. In certain embodiments, R.sup.1 is F. In certain
embodiments, R.sup.1 is Cl. In certain embodiments, R.sup.1 is Br.
In certain embodiments, R.sup.1 is I. In certain embodiments,
R.sup.1 is optionally substituted alkyl. In certain embodiments,
R.sup.1 is unsubstituted alkyl. In certain embodiments, R.sup.1 is
unsubstituted C.sub.1-6 alkyl. In certain embodiments, R.sup.1 is
methyl. In certain embodiments, R.sup.1 is substituted alkyl. In
certain embodiments, R.sup.1 is substituted C.sub.1-6alkyl.
[0216] In certain embodiments, M is O. In certain embodiments, M is
S. In certain embodiments, M is NR.sup.M, wherein R.sup.M is as
defined herein. In certain embodiments, M is NH. In certain
embodiments, M is NR.sup.M, wherein R.sup.M is optionally
substituted alkyl. In certain embodiments, M is NR.sup.M, wherein
R.sup.M is unsubstituted alkyl. In certain embodiments, M is
NCH.sub.3. In certain embodiments, M is NAc.
[0217] Compounds of any one of Formulae (I'), (II'), (I), and (II)
include Ring A attached to linker X. Compounds of any one of
Formulae (I)-(II) include Ring A attached to linker X. Ring A may
be optionally substituted monocyclic carbocyclyl, optionally
substituted monocyclic heterocyclyl, optionally substituted phenyl,
or optionally substituted monocyclic heteroaryl. In certain
embodiments, Ring A is optionally substituted monocyclic
carbocyclyl. In certain embodiments, Ring A is optionally
substituted cyclohexyl. In certain embodiments, Ring A is
optionally substituted monocyclic heterocyclyl. In certain
embodiments, Ring A is optionally substituted piperidinyl. In
certain embodiments, Ring A is optionally substituted piperizinyl.
In certain embodiments, Ring A is optionally substituted
tetrahydropyranyl. In certain embodiments, Ring A is optionally
substituted phenyl. In certain embodiments, Ring A is phenyl
substituted with only X. In certain embodiments, Ring A is
optionally substituted monocyclic heteroaryl. In certain
embodiments, Ring A is optionally substituted 5-membered
heteroaryl. In certain embodiments, Ring A is optionally
substituted 6-membered heteroaryl. In certain embodiments, Ring A
is optionally substituted pyridine. In certain embodiments, Ring A
is optionally substituted pyrimidine.
[0218] In certain embodiments, Ring A is of Formula (x-i):
##STR00034##
wherein:
[0219] each of V.sup.10, V.sup.11, V.sup.12, V.sup.13, and V.sup.14
is independently O, S, N, NR.sup.A1, C, or CR.sup.A2, as valency
permits;
[0220] each instance of R.sup.A1 is independently selected from the
group consisting of hydrogen, substituted or unsubstituted acyl,
substituted or unsubstituted alkyl, substituted or unsubstituted
alkenyl, substituted or unsubstituted alkynyl, substituted or
unsubstituted carbocyclyl, substituted or unsubstituted
heterocyclyl, substituted or unsubstituted aryl, substituted or
unsubstituted heteroaryl, and a nitrogen protecting group;
[0221] each instance of R.sup.A2 is independently selected from the
group consisting of hydrogen, halogen, substituted or unsubstituted
acyl, substituted or unsubstituted alkyl, substituted or
unsubstituted alkenyl, substituted or unsubstituted alkynyl,
substituted or unsubstituted carbocyclyl, substituted or
unsubstituted heterocyclyl, substituted or unsubstituted aryl,
substituted or unsubstituted heteroaryl, --CN, --OR.sup.A2a,
--N(R.sup.A2a).sub.2, and --SR.sup.A2a; and
[0222] each occurrence of R.sup.A2a is independently selected from
the group consisting of hydrogen, substituted or unsubstituted
acyl, substituted or unsubstituted alkyl, substituted or
unsubstituted alkenyl, substituted or unsubstituted alkynyl,
substituted or unsubstituted carbocyclyl, substituted or
unsubstituted heterocyclyl, substituted or unsubstituted aryl,
substituted or unsubstituted heteroaryl, a nitrogen protecting
group when attached to a nitrogen atom, an oxygen protecting group
when attached to an oxygen atom, and a sulfur protecting group when
attached to a sulfur atom, or two R.sup.A2a groups are joined to
form a substituted or unsubstituted heterocyclic ring.
[0223] In certain embodiments, only one of V.sup.10, V.sup.11,
V.sup.12, V.sup.13, and V.sup.14 is selected from the group
consisting of O, S, N, and NR.sup.A1. In certain embodiments, Ring
A is of the formula:
##STR00035##
[0224] In certain embodiments, Ring A is of the formula:
##STR00036##
[0225] In certain embodiments, Ring A is of the formula:
##STR00037##
[0226] In certain embodiments, only two of V.sup.10, V.sup.11,
V.sup.12, V.sup.13, and V.sup.14 are each independently selected
from the group consisting of O, S, N, and NR.sup.A1. In certain
embodiments, Ring A is of the formula:
##STR00038##
[0227] In certain embodiments, Ring A is of the formula:
##STR00039##
[0228] In certain embodiments, Ring A is of Formula (x-ii):
##STR00040##
[0229] In certain embodiments, Ring A is of the formula:
##STR00041##
[0230] In certain embodiments, Ring A is of the formula:
##STR00042##
[0231] In certain embodiments, Ring A is of the formula:
##STR00043##
[0232] In certain embodiments, Ring A is of the formula:
##STR00044##
In certain embodiments, Ring A is of the formula:
##STR00045##
In certain embodiments, Ring A is of the formula:
##STR00046##
[0233] In certain embodiments, only three of V.sup.10, V.sup.11,
V.sup.12, V.sup.13, and V.sup.14 are each independently selected
from the group consisting of O, S, N, and NR.sup.A1. In certain
embodiments, Ring A is of the formula:
##STR00047##
[0234] In certain embodiments, Ring A is of the formula:
##STR00048##
[0235] In certain embodiments, Ring A is of the formula:
##STR00049##
[0236] In certain embodiments, only four of V.sup.10, V.sup.11,
V.sup.12, V.sup.13, and V.sup.14 are each independently selected
from the group consisting of N and NR.sup.A1. In certain
embodiments, Ring A is of the formula:
##STR00050##
[0237] In certain embodiments, Ring A may also be a substituted or
unsubstituted 6-membered heteroaryl ring. In certain embodiments,
Ring A is of the formula:
##STR00051##
wherein each of V.sup.10, V.sup.11, V.sup.12, V.sup.13, V.sup.14,
and V.sup.15 is independently N, C, or CR.sup.A2, as valency
permits, wherein R.sup.A2 is as defined herein. In certain
embodiments, only one of V.sup.10, V.sup.11, V.sup.12, V.sup.13,
V.sup.14, and V.sup.15 is N. In certain embodiments, Ring A is of
the formula:
##STR00052##
wherein k is 0, 1, 2, 3, or 4, and R.sup.A2 is as defined herein.
In certain embodiments, Ring A is of the formula:
##STR00053##
[0238] In certain embodiments, only two of V.sup.10, V.sup.11,
V.sup.12, V.sup.13, V.sup.14, and V.sup.15 are N. In certain
embodiments, Ring A is of the formula:
##STR00054##
[0239] In certain embodiments, only three of V.sup.10, V.sup.11,
V.sup.12, V.sup.13, V.sup.14, and V.sup.15 are N. In certain
embodiments, Ring A is of the formula:
##STR00055##
[0240] In certain embodiments, Ring A is of the formula:
##STR00056##
wherein R.sup.A and a1 are as defined herein.
[0241] In certain embodiments, a1 is 1; and Ring A is one of the
formulae:
##STR00057##
In certain embodiments, a1 is 2; and Ring A is one of the
formulae:
##STR00058##
In certain embodiments, a2 is 3; and R.sub.61 is one of the
formulae:
##STR00059##
In certain embodiments, a2 is 4; and R.sub.61 is one of the
formulae:
##STR00060##
In certain embodiments, a2 is 5; and R.sub.61 is of the formula
##STR00061##
[0242] In certain embodiments, Ring A is of the formula:
##STR00062##
wherein R.sup.A and a1 are as defined herein.
[0243] In certain embodiments, a1 is 1; and Ring A is one of the
formulae:
##STR00063##
In certain embodiments, a1 is 2; and Ring A is one of the
formulae:
##STR00064##
In certain embodiments, a2 is 3; and R.sub.61 is one of the
formulae:
##STR00065##
In certain embodiments, a2 is 4; and R.sub.61 is one of the
formulae:
##STR00066##
In certain embodiments, a2 is 5; and R.sub.61 is of the formula
##STR00067##
[0244] In certain embodiments, Ring A is optionally substituted
monocyclic heterocyclyl. In certain embodiments, Ring A is
optionally substituted 5-membered heterocyclyl. In certain
embodiments, Ring A is optionally substituted 6-membered
heterocyclyl. In certain embodiments, Ring A is optionally
substituted 6-membered heterocyclyl with one heteroatom selected
from the group of S, O, and N.
[0245] In certain embodiments, Ring A is of the formula:
##STR00068##
wherein H.sup.A is S, O, or NR.sup.HA; R.sup.HA is hydrogen,
optionally substituted alkyl, or a nitrogen protecting group; and
R.sup.A and a1 are as defined herein.
[0246] In certain embodiments, Ring A is of the formula:
##STR00069##
wherein H.sup.A, R.sup.A and a1 are as defined herein.
[0247] In certain embodiments, Ring A is of the formula:
##STR00070##
wherein H.sup.A, R.sup.A and a1 are as defined herein.
[0248] In certain embodiments, H.sup.A is S. In certain
embodiments, H.sup.A is O. In certain embodiments, H.sup.A is
NR.sup.HA.
[0249] In certain embodiments, Ring A is of the formula:
##STR00071##
[0250] In certain embodiments, Ring A is of the formula:
##STR00072##
wherein R.sup.HA is hydrogen, optionally substituted alkyl, or a
nitrogen protecting group; and R.sup.A and a1 are as defined
herein. In certain embodiments, Ring A is of the formula:
##STR00073##
[0251] In certain embodiments, R.sup.A is hydrogen. In certain
embodiments, R.sup.A is halogen. In certain embodiments, R.sup.A is
F. In certain embodiments, R.sup.A is Cl. In certain embodiments,
R.sup.A is Br. In certain embodiments, R.sup.A is I. In certain
embodiments, R.sup.A is optionally substituted alkyl. In certain
embodiments, R.sup.A is unsubstituted alkyl. In certain
embodiments, R.sup.A is methyl, ethyl, n-propyl, i-propyl, n-butyl,
s-butyl, t-butyl, n-pentyl, t-pentyl, neo-pentyl, i-pentyl,
s-pentyl, or 3-pentyl. In certain embodiments, R.sup.A is t-butyl.
In certain embodiments, R.sup.A is substituted alkyl. In certain
embodiments, R.sup.A is haloalkyl. In certain embodiments, R.sup.A
is --CF.sub.3, --CHF.sub.2, or --CH.sub.2F. In certain embodiments,
R.sup.A is optionally substituted alkenyl. In certain embodiments,
R.sup.A is optionally substituted alkynyl. In certain embodiments,
R.sup.A is optionally carbocyclyl. In certain embodiments, R.sup.A
is optionally substituted heterocyclyl. In certain embodiments,
R.sup.A is optionally substituted aryl. In certain embodiments,
R.sup.A is optionally substituted heteroaryl. In certain
embodiments, R.sup.A is --OR.sup.a, wherein R.sup.a is as defined
herein. In certain embodiments, R.sup.A is --OH. In certain
embodiments, R.sup.A is --OR.sup.a, wherein R.sup.a is optionally
substituted alkyl or an oxygen protecting group. In certain
embodiments, R.sup.A is --OR.sup.a, wherein R.sup.a is
unsubstituted alkyl. In certain embodiments, R.sup.A is
--OCH.sub.3. In certain embodiments, R.sup.A is --N(R.sup.a).sub.2,
wherein R.sup.a is as defined herein. In certain embodiments,
R.sup.A is --NH.sub.2. In certain embodiments, R.sup.A is
--N(R.sup.a).sub.2, wherein each instance of R.sup.a is optionally
substituted alkyl or a nitrogen protecting group. In certain
embodiments, R.sup.A is --N(CH.sub.3).sub.2. In certain
embodiments, R.sup.A is --NHR.sup.a, wherein R.sup.a is optionally
substituted alkyl or a nitrogen protecting group. In certain
embodiments, R.sup.A is --NHCH.sub.3.
[0252] In certain embodiments, at least one instance of R.sup.A1 is
H (hydrogen). In certain embodiments, at least one instance of
R.sup.A1 is halogen. In certain embodiments, at least one instance
of R.sup.A1 is F (fluorine). In certain embodiments, at least one
instance of R.sup.A1 is Cl (chlorine). In certain embodiments, at
least one instance of R.sup.A1 is Br (bromine). In certain
embodiments, at least one instance of R.sup.A1 is I (iodine). In
certain embodiments, at least one instance of R.sup.A1 is
substituted acyl. In certain embodiments, at least one instance of
R.sup.A1 is unsubstituted acyl. In certain embodiments, at least
one instance of R.sup.A1 is acetyl. In certain embodiments, at
least one instance of R.sup.A1 is substituted acetyl. In certain
embodiments, at least one instance of R.sup.A1 is substituted
alkyl. In certain embodiments, at least one instance of R.sup.A1 is
unsubstituted alkyl. In certain embodiments, at least one instance
of R.sup.A1 is C.sub.1-6 alkyl. In certain embodiments, at least
one instance of R.sup.A1 is methyl. In certain embodiments, at
least one instance of R.sup.A1 is ethyl. In certain embodiments, at
least one instance of R.sup.A1 is propyl. In certain embodiments,
at least one instance of R.sup.A1 is butyl. In certain embodiments,
at least one instance of R.sup.A1 is substituted alkenyl. In
certain embodiments, at least one instance of R.sup.A1 is
unsubstituted alkenyl. In certain embodiments, at least one
instance of R.sup.A1 is vinyl. In certain embodiments, at least one
instance of R.sup.A1 is substituted alkynyl. In certain
embodiments, at least one instance of R.sup.A1 is unsubstituted
alkynyl. In certain embodiments, at least one instance of R.sup.A1
is ethynyl. In certain embodiments, at least one instance of
R.sup.A1 is substituted carbocyclyl. In certain embodiments, at
least one instance of R.sup.A1 is unsubstituted carbocyclyl. In
certain embodiments, at least one instance of R.sup.A1 is
substituted heterocyclyl. In certain embodiments, at least one
instance of R.sup.A1 is unsubstituted heterocyclyl. In certain
embodiments, at least one instance of R.sup.A1 is substituted aryl.
In certain embodiments, at least one instance of R.sup.A1 is
unsubstituted aryl. In certain embodiments, at least one instance
of R.sup.A1 is substituted phenyl. In certain embodiments, at least
one instance of R.sup.A1 is unsubstituted phenyl. In certain
embodiments, at least one instance of R.sup.A1 is substituted
heteroaryl. In certain embodiments, at least one instance of
R.sup.A1 is unsubstituted heteroaryl. In certain embodiments, at
least one instance of R.sup.A1 is substituted pyridyl. In certain
embodiments, at least one instance of R.sup.A1 is unsubstituted
pyridyl. In certain embodiments, at least one instance of R.sup.A1
is a nitrogen protecting group. In certain embodiments, at least
one instance of R.sup.A1 is BOC.
[0253] In certain embodiments, at least one R.sup.A1 is hydrogen,
substituted or unsubstituted C.sub.1-6 alkyl, or a nitrogen
protecting group. In certain embodiments, all instances of R.sup.A1
are each independently hydrogen, substituted or unsubstituted
C.sub.1-6 alkyl, or a nitrogen protecting group. In certain
embodiments, all instances of R.sup.A1 are hydrogen.
[0254] In certain embodiments, at least one R.sup.A2 is H. In
certain embodiments, at least one R.sup.A2 is halogen. In certain
embodiments, at least one R.sup.A2 is F. In certain embodiments, at
least one R.sup.A2 is Cl. In certain embodiments, at least one
R.sup.A2 is Br. In certain embodiments, at least one R.sup.A2 is I
(iodine). In certain embodiments, at least one R.sup.A2 is
substituted acyl. In certain embodiments, at least one R.sup.A2 is
unsubstituted acyl. In certain embodiments, at least one R.sup.A2
is acetyl. In certain embodiments, at least one R.sup.A2 is
substituted acetyl. In certain embodiments, at least one R.sup.A2
is substituted alkyl. In certain embodiments, at least one R.sup.A2
is unsubstituted alkyl. In certain embodiments, at least one
R.sup.A2 is C.sub.1-6 alkyl. In certain embodiments, at least one
R.sup.A2 is methyl. In certain embodiments, at least one R.sup.A2
is ethyl. In certain embodiments, at least one R.sup.A2 is propyl.
In certain embodiments, at least one R.sup.A2 is butyl. In certain
embodiments, at least one R.sup.A2 is substituted alkenyl. In
certain embodiments, at least one R.sup.A2 is unsubstituted
alkenyl. In certain embodiments, at least one R.sup.A2 is vinyl. In
certain embodiments, at least one R.sup.A2 is substituted alkynyl.
In certain embodiments, at least one R.sup.A2 is unsubstituted
alkynyl. In certain embodiments, at least one R.sup.A2 is ethynyl.
In certain embodiments, at least one R.sup.A2 is substituted
carbocyclyl. In certain embodiments, at least one R.sup.A2 is
unsubstituted carbocyclyl. In certain embodiments, at least one
R.sup.A2 is substituted heterocyclyl. In certain embodiments, at
least one R.sup.A2 is unsubstituted heterocyclyl. In certain
embodiments, at least one R.sup.A2 is substituted aryl. In certain
embodiments, at least one R.sup.A2 is unsubstituted aryl. In
certain embodiments, at least one R.sup.A2 is substituted phenyl.
In certain embodiments, at least one R.sup.A2 is unsubstituted
phenyl. In certain embodiments, at least one R.sup.A2 is
substituted heteroaryl. In certain embodiments, at least one
R.sup.A2 is unsubstituted heteroaryl. In certain embodiments, at
least one R.sup.A2 is substituted pyridyl. In certain embodiments,
at least one R.sup.A2 is unsubstituted pyridyl. In certain
embodiments, at least one R.sup.A2 is --OR.sup.A2, wherein
R.sup.A2a is as defined herein. In certain embodiments, at least
one R.sup.A2 is --OR.sup.A2a, wherein R.sup.A2a is hydrogen. In
certain embodiments, at least one R.sup.A2 is --OR.sup.A2a, wherein
R.sup.A2a is substituted or unsubstituted C.sub.1-6 alkyl. In
certain embodiments, at least one R.sup.A2 is --OR.sup.A2a, wherein
R.sup.A2a is unsubstituted C.sub.1-6 alkyl. In certain embodiments,
at least one R.sup.A2 is --OCH.sub.3. In certain embodiments, at
least one R.sup.A2 is --N(R.sup.A2a).sub.2. In certain embodiments,
at least one R.sup.A2 is --SR.sup.A2a. In certain embodiments, all
instances of R.sup.A2 are hydrogen.
[0255] In certain embodiments, all R.sup.A1 and R.sup.A2 are
hydrogen. In certain embodiments, R.sup.A1 is hydrogen; and at
least one R.sup.A2 is substituted or unsubstituted alkyl. In
certain embodiments, R.sup.A1 is hydrogen; and at least one
R.sup.A2 is unsubstituted alkyl. In certain embodiments, R.sup.A1
is hydrogen; and at least one R.sup.A2 is methyl, ethyl, or
n-propyl. In certain embodiments, R.sup.A1 is hydrogen; and at
least one R.sup.A2 is --OR.sup.A2a, wherein R.sup.A2a is as defined
herein. In certain embodiments, R.sup.A1 is hydrogen; and at least
one R.sup.A2 is --OR.sup.A2a, wherein R.sup.A2a is substituted or
unsubstituted C.sub.1-6 alkyl. In certain embodiments, R.sup.A1 is
hydrogen; and at least one R.sup.A2 is --OR.sup.A2a, wherein
R.sup.A2a is unsubstituted C.sub.1-6 alkyl. In certain embodiments,
R.sup.A1 is hydrogen; and at least one R.sup.A2 is --OCH.sub.3.
[0256] In certain embodiments, Ring A is of the formula:
##STR00074##
[0257] In certain embodiments, Ring A is of the formula:
##STR00075##
[0258] In certain embodiments, Ring A is of the formula:
##STR00076##
In certain embodiments, Ring A is of one of the formula:
##STR00077##
[0259] In certain embodiments, Ring A is of the formula:
##STR00078##
[0260] Compounds of any one of Formulae (I'), (II'), (I), and (II)
include Ring B between linker L.sup.1 and linker L.sup.2. Compounds
of any one of Formulae (I)-(II) include Ring B between linker
L.sup.1 and linker L.sup.2. Ring B may be optionally substituted
monocyclic carbocyclyl, optionally substituted monocyclic
heterocyclyl, optionally substituted phenyl, or optionally
substituted monocyclic heteroaryl. In certain embodiments, Ring B
is optionally substituted monocyclic carbocyclyl. In certain
embodiments, Ring B is optionally substituted monocyclic
heterocyclyl. In certain embodiments, Ring B is optionally
substituted pyrrolidinyl. In certain embodiments, Ring B is
optionally substituted phenyl. In certain embodiments, Ring B is
optionally substituted monocyclic heteroaryl. In certain
embodiments, Ring B is phenyl substituted with only L.sup.1 and
L.sup.2. In certain embodiments, Ring B is optionally substituted
cyclohexyl. In certain embodiments, Ring B is optionally
substituted piperidinyl. In certain embodiments, Ring B is
optionally substituted piperizinyl. In certain embodiments, Ring B
is optionally substituted pyridinyl. In certain embodiments, Ring B
is optionally substituted pyrimidinyl.
[0261] In certain embodiments of Formulae (I'), (II'), (I), and
(II), Ring B is
##STR00079##
wherein each ring atom is optionally substituted. In certain
embodiments, Ring B is
##STR00080##
wherein each ring atom is optionally substituted. In certain
embodiments, Ring B is
##STR00081##
wherein each ring atom is optionally substituted. In certain
embodiments, Ring B is
##STR00082##
wherein each ring atom is optionally substituted, and L.sup.1 and
L.sup.2 may attach to Ring B at either indicated position. In
certain embodiments, Ring B is
##STR00083##
wherein each ring atom is optionally substituted, and L.sup.2 and
R.sup.2 may attach to Ring B at either indicated position. In
certain embodiments, Ring B is
##STR00084##
wherein each ring atom is optionally substituted, and L.sup.2 and
R.sup.2 is attached to Ring B at either position indicated. In
certain embodiments, Ring B is
##STR00085##
wherein each ring atom is optionally substituted, and L.sup.1 and
L.sup.2 may attach to Ring B at either indicated position. In
certain embodiments, Ring B is
##STR00086##
wherein each ring atom is optionally substituted, and L.sup.1 and
L.sup.2 may attach to Ring B at either indicated position.
[0262] Compounds of Formula (I') and (I) include Ring C between
linker L.sup.1 and R.sup.2. Ring C may be optionally substituted
monocyclic carbocyclyl, optionally substituted monocyclic
heterocyclyl, optionally substituted phenyl, or optionally
substituted monocyclic heteroaryl. In certain embodiments, Ring C
is optionally substituted monocyclic carbocyclyl. In certain
embodiments, Ring C is optionally substituted monocyclic
heterocyclyl. In certain embodiments, Ring C is optionally
substituted monocyclic aryl. In certain embodiments, Ring C is
optionally substituted phenyl. In certain embodiments, Ring C is
optionally substituted bicyclic aryl. In certain embodiments, Ring
C is optionally substituted 2,3-dihydro-1H-indene. In certain
embodiments, Ring C is optionally substituted naphthalene. In
certain embodiments, Ring C is:
##STR00087##
In certain embodiments, Ring C is:
##STR00088##
In certain embodiments, Ring C is optionally substituted monocyclic
heteroaryl. In certain embodiments, Ring C is phenyl substituted
with only L.sup.1 and R.sup.2. In certain embodiments, Ring C is
optionally substituted cyclohexyl. In certain embodiments, Ring C
is optionally substituted pyridinone. In certain embodiments, Ring
C is:
##STR00089##
In certain embodiments, Ring C is:
##STR00090##
In certain embodiments, Ring C is optionally substituted
piperidinyl. In certain embodiments, Ring C is optionally
substituted piperizinyl. In certain embodiments, Ring C is
optionally substituted pyridinyl. In certain embodiments, Ring C is
optionally substituted pyrimidinyl. In certain embodiments, Ring C
is optionally substituted bicyclic heteroaryl. In certain
embodiments, Ring C is optionally substituted indolyl. In certain
embodiments, Ring C is optionally substituted indolinyl. In certain
embodiments, Ring C is:
##STR00091##
In certain embodiments, Ring C is:
##STR00092##
[0263] In certain embodiments, for Formula (I'), L.sup.1 is
--CH.sub.2--. In certain embodiments, for Formula (I'), L.sup.1 is
.sup.lc--S(.dbd.O).sub.2--.sup.lb. In certain embodiments, L.sup.1
is --O--. In certain embodiments, L.sup.1 is --S--. In certain
embodiments, L.sup.1 is --NR.sup.L1--. In certain embodiments,
L.sup.1 is --C(.dbd.O)--. In certain embodiments, L.sup.1 is
--NR.sup.L1--. In certain embodiments, L.sup.1 is
.sup.lc--OC(.dbd.O)--.sup.lb. In certain embodiments, L.sup.1 is
.sup.lc--C(.dbd.O)O--.sup.lb. In certain embodiments, L.sup.1 is
.sup.lc--NR.sup.L1C(.dbd.O)--.sup.lb. In certain embodiments,
L.sup.1 is .sup.lc--C(O)NR.sup.L1--.sup.lb. As used herein, .sup.lb
indicates the point of attachment is to Ring B; and .sup.lc
indicates the point of attachment is to Ring C.
[0264] In certain embodiments, R.sup.L1 is hydrogen. In certain
embodiments, R.sup.L1 is optionally substituted C.sub.1-6 alkyl. In
certain embodiments, R.sup.L1 is unsubstituted C.sub.1-6 alkyl. In
certain embodiments, R.sup.L1 is methyl. In certain embodiments,
R.sup.L1 is substituted C.sub.1-6 alkyl. In certain embodiments,
R.sup.L1 is a nitrogen protecting group.
[0265] In certain embodiments, L.sup.2 is --O--. In certain
embodiments, L.sup.2 is --S--. In certain embodiments, L.sup.2 is
--NR.sup.L2--. In certain embodiments, L.sup.2 is
.sup.lb--NR.sup.L2C(.dbd.O)--.sup.lm. In certain embodiments,
L.sup.2 is .sup.lb--C(.dbd.O)NR.sup.L2--.sup.lm. As used herein,
.sup.lb indicates the point of attachment is to Ring B; and .sup.lm
indicates the point of attachment is to the heteroaryl ring with
M.
[0266] In certain embodiments, R.sup.L2 is hydrogen. In certain
embodiments, R.sup.L2 is optionally substituted C.sub.1-6 alkyl. In
certain embodiments, R.sup.L2 is unsubstituted C.sub.1-6 alkyl. In
certain embodiments, R.sup.L2 is methyl. In certain embodiments,
R.sub.L2 is substituted C.sub.1-6 alkyl. In certain embodiments,
R.sup.L2 is a nitrogen protecting group.
[0267] In certain embodiments, for Formula (II'), X is a bond. In
certain embodiments, for Formula (I'), X is
.sup.xm--CH.sub.2CH.sub.2--.sup.xa. In certain embodiments, for
Formula (I'), X is .sup.xm--CH.dbd.CH--.sup.xa. In certain
embodiments, for Formula (I'), X is
.sup.xm--CH.sub.2--NR.sup.LX--.sup.xa. In certain embodiments, for
Formula (I'), X is .sup.xm--CH.sub.2--NH--.sup.xa. In certain
embodiments, for Formula (I'), X is
.sup.xm--CH.sub.2--O--CH.sub.2--.sup.xa. In certain embodiments,
for Formula (I'), X is .sup.xa--CH.sub.2--NH--CH.sub.2--.sup.xa. In
certain embodiments, X is --O--. In certain embodiments, X is
--S--. In certain embodiments, X is
.sup.xm--S--C(.dbd.O)CH.sub.2--.sup.xa. In certain embodiments, X
is --NR.sup.LX--. In certain embodiments, X is --O--CH.sub.2--. In
certain embodiments, X is --S--CH.sub.2--. In certain embodiments,
X is --NR.sup.LX--CH.sub.2--. In certain embodiments, X is
--NH--CH.sub.2--.
[0268] In certain embodiments, a1 is 1. In certain embodiments, a1
is 2. In certain embodiments, a1 is 3.
[0269] In certain embodiments, b1 is 1. In certain embodiments, b1
is 2. In certain embodiments, b1 is 3.
[0270] In certain embodiments, c1 is 1. In certain embodiments, c1
is 2. In certain embodiments, c1 is 3.
[0271] In certain embodiments, R.sup.B is optionally substituted
alkyl. In certain embodiments, R.sup.B is optionally substituted
C.sub.1-C.sub.6 alkyl. In certain embodiments, R.sup.B is
unsubstituted C.sub.1-C.sub.6 alkyl. In certain embodiments,
R.sup.B is methyl or ethyl. In certain embodiments, R.sup.B is
substituted C.sub.1-C.sub.6 alkyl. In certain embodiments, R.sup.B
is hydroxy C.sub.1-C.sub.6 alkyl. In certain embodiments, R.sup.B
is --CH.sub.2OH. In certain embodiments, R.sup.B is
--CH.sub.2CH.sub.2OH. In certain embodiments, R.sup.B is
--N(R.sup.a).sub.2, wherein R.sup.a is as defined herein. In
certain embodiments, R.sup.B is --NHR.sup.a, wherein R.sup.a is as
defined herein. In certain embodiments, R.sup.B is --NHR.sup.a,
wherein R.sup.a is hydrogen or optionally substituted
C.sub.1-C.sub.6 alkyl. In certain embodiments, R.sup.B is
--NH.sub.2. In certain embodiments, R.sup.B is --NHR.sup.a, wherein
R.sup.a is optionally substituted C.sub.1-C.sub.6 alkyl. In certain
embodiments, R.sup.B is --NHR.sup.a, wherein R.sup.a is
unsubstituted C.sub.1-C.sub.6 alkyl. In certain embodiments,
R.sup.B is --NHR.sup.a, wherein R.sup.a is methyl or ethyl. In
certain embodiments, R.sup.B is --NHCH.sub.3. In certain
embodiments, R.sup.B is --NHR.sup.a, wherein R.sup.a is a nitrogen
protecting group. In certain embodiments, R.sup.B is
--N(CH.sub.3)R.sup.a, wherein R.sup.a is optionally substituted
C.sub.1-C.sub.6 alkyl. In certain embodiments, R.sup.B is
--N(CH.sub.3)R.sup.a, wherein R.sup.a is unsubstituted
C.sub.1-C.sub.6 alkyl. In certain embodiments, R.sup.B is
--N(CH.sub.3)R.sup.a, wherein R.sup.a is methyl or ethyl. In
certain embodiments, R.sup.B is --N(CH.sub.3).sub.2. In certain
embodiments, R.sup.B is --N(CH.sub.3)R.sup.a, wherein R.sup.a is a
nitrogen protecting group.
[0272] In certain embodiments, R.sup.C is optionally substituted
alkyl. In certain embodiments, R.sup.C is optionally substituted
C.sub.1-C.sub.6 alkyl. In certain embodiments, R.sup.C is
unsubstituted C.sub.1-C.sub.6 alkyl. In certain embodiments,
R.sup.C is methyl or ethyl. In certain embodiments, R.sup.C is
substituted C.sub.1-C.sub.6 alkyl. In certain embodiments, R.sup.C
is hydroxy C.sub.1-C.sub.6 alkyl. In certain embodiments, R.sup.C
is --CH.sub.2OH. In certain embodiments, R.sup.C is
--CH.sub.2CH.sub.2OH. In certain embodiments, R.sup.C is
--N(R.sup.a).sub.2, wherein R.sup.a is as defined herein. In
certain embodiments, R.sup.C is --NHR.sup.a, wherein R.sup.a is as
defined herein. In certain embodiments, R.sup.C is --NHR.sup.a,
wherein R.sup.a is hydrogen or optionally substituted
C.sub.1-C.sub.6 alkyl. In certain embodiments, R.sup.C is
--NH.sub.2. In certain embodiments, R.sup.C is --NHR.sup.a, wherein
R.sup.a is optionally substituted C.sub.1-C.sub.6 alkyl. In certain
embodiments, R.sup.C is --NHR.sup.a, wherein R.sup.a is
unsubstituted C.sub.1-C.sub.6 alkyl. In certain embodiments,
R.sup.C is --NHR.sup.a, wherein R.sup.a is methyl or ethyl. In
certain embodiments, R.sup.C is --NHCH.sub.3. In certain
embodiments, R.sup.C is --NHR.sup.a, wherein R.sup.a is a nitrogen
protecting group. In certain embodiments, R.sup.C is
--N(CH.sub.3)R.sup.a, wherein R.sup.a is optionally substituted
C.sub.1-C.sub.6 alkyl. In certain embodiments, R.sup.C is
--N(CH.sub.3)R.sup.a, wherein R.sup.a is unsubstituted
C.sub.1-C.sub.6 alkyl. In certain embodiments, R.sup.C is
--N(CH.sub.3)R.sup.a, wherein R.sup.a is methyl or ethyl. In
certain embodiments, R.sup.C is --N(CH.sub.3).sub.2. In certain
embodiments, R.sup.C is --N(CH.sub.3)R.sup.a, wherein R.sup.a is a
nitrogen protecting group.
[0273] Compounds of Formula (I') include R.sup.2 attached to Ring
C. Compounds of Formula (I) include R.sup.2 attached to Ring C.
Compounds of Formula (I) include R.sup.2 attached to Ring B.
Compounds of Formula (I') include R.sup.2 attached to Ring B. In
certain embodiments, R.sup.2 comprises an electrophilic moiety. In
certain embodiments, R.sup.2 comprises a Michael acceptor moiety.
The electrophilic moiety (e.g., Michael acceptor moiety) may react
with a cysteine residue of a kinase (e.g., CDK (e.g., CDK7)) to
allow for covalent attachment of the compound to the kinase. In
certain embodiments, the electrophilic moiety (e.g., Michael
acceptor moiety) may react with a cysteine residue of a kinase
(e.g., CDK (e.g., CDK7)). In certain embodiments, the electrophilic
moiety (e.g., Michael acceptor moiety) may react with the Cys312
residue of CDK7. In certain embodiments, the covalent attachment is
irreversible. In certain embodiments, the covalent attachment is
reversible.
[0274] As generally defined herein in Formulae (I'), (II'), (I),
and (II), R.sup.2 may be any one of Formulae (i-1)-(i-41). In
certain embodiments, R.sup.2 is of Formula (i-1):
##STR00093##
In certain embodiments, R.sup.2 is of Formula (i-2):
##STR00094##
In certain embodiments, R.sup.2 is of Formula (i-3):
##STR00095##
In certain embodiments, R.sup.2 is of Formula (i-4):
##STR00096##
In certain embodiments, R.sup.2 is of Formula (i-5):
##STR00097##
In certain embodiments, R.sup.2 is of Formula (i-6):
##STR00098##
In certain embodiments, R.sup.2 is of Formula (i-7):
##STR00099##
In certain embodiments, R.sup.2 is of Formula (i-8):
##STR00100##
In certain embodiments, R.sup.2 is of Formula (i-9):
##STR00101##
In certain embodiments, R.sup.2 is of Formula (i-10):
##STR00102##
In certain embodiments, R.sup.2 is of Formula (i-11):
##STR00103##
In certain embodiments, R.sup.2 is of Formula (i-12):
##STR00104##
In certain embodiments, R.sup.2 is of Formula (i-13):
##STR00105##
In certain embodiments, R.sup.2 is of Formula (i-14):
##STR00106##
In certain embodiments, R.sup.2 is of Formula (i-15):
##STR00107##
In certain embodiments, R.sup.2 is of Formula (i-16):
##STR00108##
In certain embodiments, R.sup.2 is of Formula (i-17):
##STR00109##
In certain embodiments, R.sup.2 is of Formula (i-18):
##STR00110##
In certain embodiments, R.sup.2 is of Formula (i-19):
##STR00111##
In certain embodiments, R.sup.2 is of Formula (i-20):
##STR00112##
In certain embodiments, R.sup.2 is of Formula (i-21):
##STR00113##
In certain embodiments, R.sup.2 is of Formula (i-22):
##STR00114##
In certain embodiments, R.sup.2 is of Formula (i-23):
##STR00115##
In certain embodiments, R.sup.2 is of Formula (i-24):
##STR00116##
In certain embodiments, R.sup.2 is of Formula (i-25):
##STR00117##
In certain embodiments, R.sup.2 is of Formula (i-26):
##STR00118##
In certain embodiments, R.sup.2 is of Formula (i-27):
##STR00119##
In certain embodiments, R.sup.2 is of Formula (i-28):
##STR00120##
In certain embodiments, R.sup.2 is of Formula (i-29):
##STR00121##
In certain embodiments, R.sup.2 is of Formula (i-30):
##STR00122##
In certain embodiments, R.sup.2 is of Formula (i-31):
##STR00123##
In certain embodiments, R.sup.2 is of Formula (i-32):
##STR00124##
In certain embodiments, R.sup.2 is of Formula (i-33):
##STR00125##
In certain embodiments, R.sup.2 is of Formula (i-34):
##STR00126##
In certain embodiments, R.sup.2 is of Formula (i-35):
##STR00127##
In certain embodiments, R.sup.2 is of Formula (i-36):
##STR00128##
In certain embodiments, R.sup.2 is of Formula (i-37):
##STR00129##
In certain embodiments, R.sup.2 is of Formula (i-38):
##STR00130##
In certain embodiments, R.sup.2 is of Formula (i-39)
##STR00131##
In certain embodiments, R.sup.2 is of Formula (i-40):
##STR00132##
In certain embodiments, R.sup.2 is of Formula (i-41):
##STR00133##
[0275] In certain embodiments, R.sup.2 is of Formula (i-1a):
##STR00134##
In certain embodiments, R.sup.2 is of Formula (i-1b):
##STR00135##
In certain embodiments, R.sup.2 is of Formula (i-1c):
##STR00136##
In certain embodiments, R.sup.2 is of Formula (i-1d):
##STR00137##
In certain embodiments, R.sup.2 is of Formula (i-1e):
##STR00138##
In certain embodiments, R.sup.2 is of Formula (i-1f):
##STR00139##
In certain embodiments, R.sup.2 is of Formula (i-1g)
##STR00140##
In certain embodiments, R.sup.2 is
##STR00141##
In certain embodiments, R.sup.2 is
##STR00142##
In certain embodiments, R.sup.2 is
##STR00143##
In certain embodiments, R.sup.2 is of Formula (i-1h):
##STR00144##
In certain embodiments, R.sup.2 is
##STR00145##
[0276] In certain embodiments, R.sup.2 is of Formula (i-1a):
##STR00146##
In certain embodiments, R.sup.2 is of Formula (i-1b):
##STR00147##
In certain embodiments, R.sup.2 is of Formula (i-1c):
##STR00148##
[0277] In certain embodiments, R.sup.2 is of Formula (i-18a):
##STR00149##
In certain embodiments, R.sup.2 is of Formula (i-18b):
##STR00150##
In certain embodiments, R.sup.2 is of Formula (i-18c):
##STR00151##
[0278] In certain embodiments, R.sup.2 is of Formula (i-15a):
##STR00152##
In certain embodiments, R.sup.2 is of Formula (i-15b):
##STR00153##
In certain embodiments, R.sup.2 is of Formula (i-15c):
##STR00154##
[0279] R.sup.2 may contain linker L.sup.3 or L.sup.4. In certain
embodiments, L.sup.3 is a bond. L.sup.3 is an optionally
substituted C.sub.1-4 hydrocarbon chain. In certain embodiments,
L.sup.3 is an optionally substituted C.sub.1-4 hydrocarbon chain,
wherein one or more carbon units of the hydrocarbon chain are
independently replaced with --C(.dbd.O)--, --O--, --S--,
--NR.sup.L3a--, --NR.sup.L3aC(.dbd.O)--, --C(.dbd.O)NR.sup.L3a--,
--SC(.dbd.O)--, --C(.dbd.O)S--, --OC(.dbd.O)--, --C(.dbd.O)O--,
--NR.sup.L3aC(.dbd.S)--, --C(--S)NR.sup.L3a--,
trans-CR.sup.L3b.dbd.CR.sup.L3b--, cis-CR.sup.L3b.dbd.CR.sup.L3b--,
--C.ident.C--, --S(.dbd.O)--, --S(.dbd.O)O--, --OS(.dbd.O)--,
--S(.dbd.O)NR.sup.L3a--, --NR.sup.L3aS(.dbd.O)--,
--S(.dbd.O).sub.2--, --S(.dbd.O).sub.2O--, --OS(.dbd.O).sub.2--,
--S(.dbd.O).sub.2NR.sup.L3a--, or --NR.sup.L3aS(.dbd.O).sub.2--. In
certain embodiments, L.sup.3 is an optionally substituted C.sub.1-4
hydrocarbon chain, wherein one carbon unit of the hydrocarbon chain
is replaced with --NR.sup.L3a-- (e.g., --NH--). In certain
embodiments, L.sup.3 is of the formula:
--(CH.sub.2).sub.1-4--NR.sup.L3a-- (e.g.,
--(CH.sub.2).sub.1-4--NH--) or
--NR.sup.L3a--CH.sub.2).sub.1-4--(e.g., --NH--CH.sub.2).sub.1-4--).
In certain embodiments, L.sup.3 is --NR.sup.L3a. In certain
embodiments, L.sup.3 is --NR.sup.L3a(C.dbd.O)--. In certain
embodiments, L.sup.3 is --(C.dbd.O)NR.sup.L3a--. In certain
embodiments, L.sup.3 is --NH--. In certain embodiments, L.sup.3 is
--(C.dbd.O)--. In certain embodiments, L.sup.3 is --NH(C.dbd.O)--.
In certain embodiments, L.sup.3 is --(C.dbd.O)NH--. In certain
embodiments, L.sup.3 is --O--. In certain embodiments, L.sup.3 is
--S--. In certain embodiments, L.sup.4 is a bond. In certain
embodiments, L.sup.4 is an optionally substituted C.sub.1-4
hydrocarbon chain.
[0280] Linker L.sup.3 may contain groups R.sup.L3a or R.sup.L3b. In
certain embodiments, R.sup.L3a is hydrogen. In certain embodiments,
at least one instance of R.sup.L3b is hydrogen. In certain
embodiments, each instance of R.sup.L3b is hydrogen. In certain
embodiments, at least one instance of R.sup.L3b is --Cl, --Br, or
--I. In certain embodiments, each instance of R.sup.L3b is --Cl,
--Br, or --I. In certain embodiments, at least one instance of
R.sup.L3b is --F. In certain embodiments, each instance of
R.sup.L3b is --F. In certain embodiments, at least one instance of
R.sup.L3b is optionally substituted alkyl, optionally substituted
alkenyl, optionally substituted alkynyl, optionally substituted
carbocyclyl, optionally substituted heterocyclyl, optionally
substituted aryl, or optionally substituted heteroaryl. In certain
embodiments, two R.sup.L3b groups are joined to form an optionally
substituted carbocyclic or optionally substituted heterocyclic
ring.
[0281] R.sup.2 may contain groups R.sup.E1, R.sup.E2, and/or
R.sup.E3. In certain embodiments, R.sup.E1 is hydrogen. In certain
embodiments, R.sup.E2 is hydrogen. In certain embodiments, R.sup.E3
is hydrogen. In certain embodiments, R.sup.E1 is --Cl, --Br, or
--I. In certain embodiments, R.sup.E2 is --Cl, --Br, or --I. In
certain embodiments, R.sup.E3 is --Cl, --Br, or --I. In certain
embodiments, R.sup.E1 is --F. In certain embodiments, R.sup.E2 is
--F. In certain embodiments, R.sup.E3 is --F. In certain
embodiments, R.sup.E1 is optionally substituted alkyl (e.g.,
substituted or unsubstituted C.sub.1-6 alkyl). In certain
embodiments, R.sup.E2 is optionally substituted alkyl (e.g.,
substituted or unsubstituted C.sub.1-6 alkyl). In certain
embodiments, R.sup.E3 is optionally substituted alkyl (e.g.,
substituted or unsubstituted C.sub.1-6 alkyl). In certain
embodiments, R.sup.E1 is optionally substituted alkenyl, optionally
substituted alkynyl, optionally substituted carbocyclyl, optionally
substituted heterocyclyl, optionally substituted aryl, optionally
substituted heteroaryl, --CN, --CH.sub.2OR.sup.EE,
--CH.sub.2N(R.sup.EE).sub.2, --CH.sub.2SR.sup.EE, --OR.sup.EE,
--N(R.sup.EE).sub.2, --Si(R.sup.EE).sub.3, or --SR.sup.EE. In
certain embodiments, R.sup.E2 is optionally substituted alkenyl,
optionally substituted alkynyl, optionally substituted carbocyclyl,
optionally substituted heterocyclyl, optionally substituted aryl,
optionally substituted heteroaryl, --CN, --CH.sub.2OR.sup.EE,
--CH.sub.2N(R.sup.EE).sub.2, --CH.sub.2SR.sup.EE, --OR.sup.EE,
--N(R.sup.EE).sub.2, --Si(R.sup.EE).sub.3, or --SR.sup.EE. In
certain embodiments, R.sup.E3 is optionally substituted alkenyl,
optionally substituted alkynyl, optionally substituted carbocyclyl,
optionally substituted heterocyclyl, optionally substituted aryl,
optionally substituted heteroaryl, --CN, --CH.sub.2OR.sup.EE,
--CH.sub.2N(R.sup.EE).sub.2, --CH.sub.2SR.sup.EE, --OR.sup.EE,
--N(R.sup.EE).sub.2, --Si(R.sup.EE).sub.3, or --SR.sup.EE. In
certain embodiments, R.sup.E1 is --N(R.sup.EE).sub.2. In certain
embodiments, R.sup.E2 is --N(R.sup.EE).sub.2. In certain
embodiments, R.sup.E3 is --N(R.sup.EE).sub.2. In certain
embodiments, R.sup.E1 is --N(CH.sub.3).sub.2. In certain
embodiments, R.sup.E2 is --N(CH.sub.3).sub.2. In certain
embodiments, R.sup.E3 is --N(CH.sub.3).sub.2. In certain
embodiments, R.sup.E1 is --CH.sub.2N(R.sup.EE).sub.2. In certain
embodiments, R.sup.E2 is --CH.sub.2N(R.sup.EE).sub.2. In certain
embodiments, R.sup.E3 is --CH.sub.2N(R.sup.EE).sub.2. In certain
embodiments, R.sup.E1 is --CH.sub.2N(CH.sub.3).sub.2. In certain
embodiments, R.sup.E2 is --CH.sub.2N(CH.sub.3).sub.2. In certain
embodiments, R.sup.E3 is --CH.sub.2N(CH.sub.3).sub.2. In certain
embodiments, R.sup.E1 is --CN. In certain embodiments, R.sup.E2 is
--CN. In certain embodiments, R.sup.E3 is --CN.
[0282] In certain embodiments, R.sup.E1 and R.sup.E3 are joined to
form an optionally substituted carbocyclic ring. In certain
embodiments, R.sup.E1 and R.sup.E3 are joined to form an optionally
substituted heterocyclic ring. In certain embodiments, R.sup.E2 and
R.sup.E3 are joined to form an optionally substituted carbocyclic
ring. In certain embodiments, R.sup.E2 and R.sup.E3 are joined to
form an optionally substituted heterocyclic ring. In certain
embodiments, R.sup.E1 and R.sup.E2 are joined to form an optionally
substituted carbocyclic ring. In certain embodiments, R.sup.E1 and
R.sup.E2 are joined to form an optionally substituted heterocyclic
ring.
[0283] R.sup.2 may contain group R.sup.E4, where R.sup.E4 is a
leaving group. In certain embodiments, R.sup.E4 is --Cl, --Br, or
--I. In certain embodiments, R.sup.E4 is --F. In certain
embodiments, R.sup.E4 is --OS(.dbd.O)R.sup.E4a or
--OS(.dbd.O).sub.2R.sup.E4a, wherein R.sup.E4a is substituted or
unsubstituted alkyl, substituted or unsubstituted alkenyl,
substituted or unsubstituted alkynyl, substituted or unsubstituted
carbocyclyl, substituted or unsubstituted heterocyclyl, substituted
or unsubstituted aryl, or substituted or unsubstituted heteroaryl.
In certain embodiments, R.sup.E4 is --OR.sup.E4a. In certain
embodiments, R.sup.E4 is --OMs, --OTf --OTs, --OBs, or
2-nitrobenzenesulfonyloxy. In certain embodiments, R.sup.E4 is
--OR.sup.E4a. In certain embodiments, R.sup.E4 is --OMe,
--OCF.sub.3, or --OPh. In certain embodiments, R.sup.E4 is
--OC(.dbd.O)R.sup.E4a. In certain embodiments, R.sup.E4 is
--OC(.dbd.O)Me, --OC(.dbd.O)CF.sub.3, --OC(.dbd.O)Ph, or
--OC(.dbd.O)Cl. In certain embodiments, R.sup.E4 is
--OC(O)OR.sup.E4a. In certain embodiments, R.sup.E4 is
--OC(.dbd.O)OMe or --OC(.dbd.O)O(t-Bu).
[0284] R.sup.2 may contain group R.sup.E5, where R.sup.E5 is a
halogen. In certain embodiments, R.sup.E5 is --Cl, --Br, or --I. In
certain embodiments, R.sup.E5 is --F.
[0285] R.sup.2 may contain group R.sup.E6. In certain embodiments,
R.sup.E6 is hydrogen. In certain embodiments, R.sup.E6 is
substituted or unsubstituted C.sub.1-C.sub.6 alkyl. In certain
embodiments, R.sup.E6 is a nitrogen protecting group.
[0286] In certain embodiments, a is 1. In certain embodiments, a is
2.
[0287] In certain embodiments, z is 0. In certain embodiments, z is
1. In certain embodiments, z is 2. In certain embodiments, z is 3,
4, 5, or 6.
[0288] R.sup.2 may contain group Y. In certain embodiments, Y is O.
In certain embodiments, Y is S. In certain embodiments, Y is
NR.sup.E7. In certain embodiments, Y is NH. In certain embodiments,
a compound of Formula (I') is of Formula (I).
[0289] In certain embodiments, the compound of Formula (I) is the
formula:
##STR00155##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof.
[0290] In certain embodiments, the compound of Formula (I) is the
formula:
##STR00156##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof.
[0291] In certain embodiments, the compound of Formula (I) is the
formula:
##STR00157##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof.
[0292] In certain embodiments, a compound of Formula (I) is of
Formula (I-i):
##STR00158##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof, wherein L.sup.1, L.sup.2, X, Ring
A, Ring C, R.sup.1, R.sup.2, R.sup.A, R.sup.B, R.sup.C, a1, b1, and
c1 are as defined herein.
[0293] In certain embodiments, a compound of Formula (I) is of
Formula (I-ii):
##STR00159##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof wherein L.sup.1, L.sup.2, X, Ring A,
Ring C, R.sup.1, R.sup.2, R.sup.A, R.sup.B, R.sup.C, a1, b1, and c1
are as defined herein.
[0294] In certain embodiments, a compound of Formula (I) is of
Formula (I-ii-a):
##STR00160##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof, wherein L.sup.1, L.sup.2, X, Ring
A, R.sup.1, R.sup.2, R.sup.A, R.sup.B, R.sup.C, a1, b1, and c1 are
as defined herein.
[0295] In certain embodiments, a compound of Formula (I) is of the
formula:
##STR00161##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof wherein L.sup.1, L.sup.2, X,
R.sup.1, R.sup.2, R.sup.A, R.sup.B, R.sup.C, a1, b1, and c1 are as
defined herein.
[0296] In certain embodiments, a compound of Formula (I) is of the
formula:
##STR00162##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof, wherein L.sup.1, L.sup.2, X,
R.sup.1, R.sup.2, R.sup.A, R.sup.B, R.sup.C, a1, b1, and c1 are as
defined herein.
[0297] In certain embodiments, a compound of Formula (I) is of the
formula:
##STR00163##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof, wherein L.sup.1, L.sup.2, X,
R.sup.1, R.sup.2, R.sup.A, R.sup.B, R.sup.C, a1, b1, and c1 are as
defined herein.
[0298] In certain embodiments, a compound of Formula (I) is of the
formula:
##STR00164##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof, wherein L.sup.1, L.sup.2, X,
R.sup.1, R.sup.2, R.sup.A, R.sup.B, R.sup.C, a1, b1, and c1 are as
defined herein.
[0299] In certain embodiments, a compound of Formula (I) is of the
formula:
##STR00165##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof, wherein L.sup.1, L.sup.2, X,
R.sup.1, R.sup.2, R.sup.A, R.sup.B, R.sup.C, a1, b1, and c1 are as
defined herein.
[0300] In certain embodiments, a compound of Formula (I) is of the
formula:
##STR00166##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof wherein L.sup.1, L.sup.2, X,
R.sup.1, R.sup.2, R.sup.A, R.sup.B, R.sup.C, a1, b1, and c1 are as
defined herein.
[0301] In certain embodiments, a compound of Formula (I) is of
Formula (I-ii-b):
##STR00167##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof, wherein L.sup.1, L.sup.2, X, Ring
A, R.sup.1, R.sup.2, R.sup.A, R.sup.B, R.sup.C, a1, b1, and c1 are
as defined herein.
[0302] In certain embodiments, a compound of Formula (I) is of the
formula:
##STR00168##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof, wherein L.sup.1, L.sup.2, X,
R.sup.1, R.sup.2, R.sup.A, R.sup.B, R.sup.C, a1, b1, and c1 are as
defined herein.
[0303] In certain embodiments, a compound of Formula (I) is of
Formula (I-ii-c):
##STR00169##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof, wherein L.sup.1, L.sup.2, X, Ring
A, R.sup.1, R.sup.2, R.sup.A, R.sup.B, R.sup.C, a1, b1, and c1 are
as defined herein.
[0304] In certain embodiments, a compound of Formula (I) is of the
formula:
##STR00170##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof, wherein L.sup.1, L.sup.2, X,
R.sup.1, R.sup.2, R.sup.A, R.sup.B, R.sup.C, a1, b1, and c1 are as
defined herein.
[0305] In certain embodiments, a compound of Formula (I) is of
Formula (I-ii-A):
##STR00171##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof, wherein L.sup.1, L.sup.2, X,
R.sup.1, R.sup.2, R.sup.A, R.sup.B, R.sup.C, b1, and c1 are as
defined herein.
[0306] In certain embodiments, a compound of Formula (I) is of
Formula (I-ii):
##STR00172##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof; wherein L.sup.1, L.sup.2, X, Ring
A, Ring C, R.sup.1, R.sup.2, R.sup.A, R.sup.B, R.sup.C, a1, b1, and
c1 are as defined herein.
[0307] In certain embodiments, a compound of Formula (I) is of
Formula (I-iv):
##STR00173##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof, wherein L.sup.1, L.sup.2, X, Ring
A, Ring C, R.sup.1, R.sup.2, R.sup.A, R.sup.B, R.sup.C, a1, b1, and
c1 are as defined herein.
[0308] In certain embodiments, a compound of Formula (I) is of
Formula (I-iv-a):
##STR00174##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof, wherein L.sup.1, L.sup.2, X, Ring
A, R.sup.1, R.sup.2, R.sup.A, R.sup.B, R.sup.C, a1, b1, and c1 are
as defined herein.
[0309] In certain embodiments, a compound of Formula (I) is of the
formula:
##STR00175##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof wherein L.sup.1, L.sup.2, X,
R.sup.1, R.sup.2, R.sup.A, R.sup.B, R.sup.C, a1, b1, and c1 are as
defined herein.
[0310] In certain embodiments, a compound of Formula (I) is of
Formula (I-iv-b):
##STR00176##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof, wherein L.sup.1, L.sup.2, X, Ring
A, R.sup.1, R.sup.2, R.sup.A, R.sup.B, R.sup.C, a1, b1, and c1 are
as defined herein.
[0311] In certain embodiments, a compound of Formula (I) is of the
formula:
##STR00177##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof, wherein L.sup.1, L.sup.2, X,
R.sup.1, R.sup.2, R.sup.A, R.sup.B, R.sup.C, a1, b1, and c1 are as
defined herein.
[0312] In certain embodiments, a compound of Formula (I) is of
Formula (I-v):
##STR00178##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof wherein L.sup.1, L.sup.2, X, Ring A,
Ring C, R.sup.1, R.sup.2, R.sup.A, R.sup.B, R.sup.C, a1, b1, and c1
are as defined herein.
[0313] In certain embodiments, a compound of Formula (I) is of
Formula (I-v-a):
##STR00179##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof, wherein L.sup.1, L.sup.2, X, Ring
A, R.sup.1, R.sup.2, R.sup.A, R.sup.B, R.sup.C, a1, b1, and c1 are
as defined herein.
[0314] In certain embodiments, a compound of Formula (I) is of the
formula:
##STR00180##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof, wherein L.sup.1, L.sup.2, X,
R.sup.1, R.sup.2, R.sup.A, R.sup.B, R.sup.C, a1, b1, and c1 are as
defined herein.
[0315] In certain embodiments, a compound of Formula (I) is of
Formula (I-v-b):
##STR00181##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof wherein L.sup.1, L.sup.2, X, Ring A,
R.sup.1, R.sup.2, R.sup.A, R.sup.B, R.sup.C, a1, b1, and c1 are as
defined herein.
[0316] In certain embodiments, a compound of Formula (I) is of the
formula:
##STR00182##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof, wherein L.sup.1, L.sup.2, X,
R.sup.1, R.sup.2, R.sup.A, R.sup.B, R.sup.C, a1, b1, and c1 are as
defined herein.
[0317] In certain embodiments, a compound of Formula (I') is of
Formula (I'-A):
##STR00183##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof, wherein L.sup.1, L.sup.2, X, Ring
A, R.sup.1, R.sup.2, R.sup.A, R.sup.B, R.sup.C, a1, b1, and c1 are
as defined herein.
[0318] In certain embodiments, a compound of Formula (I') is of the
formula:
##STR00184##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof, wherein L.sup.1, L.sup.2, X,
R.sup.1, R.sup.2, R.sup.A, R.sup.B, R.sup.C, b1, and c1 are as
defined herein.
[0319] In certain embodiments, a compound of Formula (I') is of
Formula (I'-i):
##STR00185##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof; wherein L.sup.1, L.sup.2, X, Ring
A, R.sup.1, R.sup.2, R.sup.A, R.sup.B, R.sup.C, a1, b1, and c1 are
as defined herein.
[0320] In certain embodiments, a compound of Formula (I') is of
Formula (I'-ii):
##STR00186##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof; wherein L.sup.1, L.sup.2, X, Ring
A, R.sup.1, R.sup.2, R.sup.A, R.sup.B, R.sup.C, a1, b1, and c1 are
as defined herein.
[0321] In certain embodiments, a compound of Formula (I') is of
Formula (I'-iii):
##STR00187##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof; wherein L.sup.1, L.sup.2, X, Ring
A, R.sup.1, R.sup.2, R.sup.A, R.sup.B, R.sup.C, a1, b1, and c1 are
as defined herein.
[0322] In certain embodiments, a compound of Formula (I') is of the
formula:
##STR00188##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof, wherein L.sup.1, L.sup.2, X,
R.sup.1, R.sup.2, R.sup.A, R.sup.B, R.sup.C, b1, and c1 are as
defined herein.
[0323] In certain embodiments, a compound of Formula (II') is of
Formula (II).
[0324] In certain embodiments, the compound of Formula (II) is the
formula:
##STR00189##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof.
[0325] In certain embodiments, the compound of Formula (II) is the
formula:
##STR00190##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof.
[0326] In certain embodiments, the compound of Formula (II) is the
formula:
##STR00191##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof.
[0327] In certain embodiments, a compound of Formula (II) is of
Formula (II-i):
##STR00192##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof, wherein L.sup.2, X, Ring A,
R.sup.1, R.sup.2, R.sup.A, R.sup.B, a1, and b1 are as defined
herein.
[0328] In certain embodiments, a compound of Formula (II) is of the
formula:
##STR00193##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof.
[0329] In certain embodiments, a compound of Formula (II) is of the
formula:
##STR00194##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof.
[0330] In certain embodiments, a compound of Formula (II') is of
Formula (II'-i):
##STR00195##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof, wherein L.sup.2, X, Ring A,
R.sup.1, R.sup.2, R.sup.A, R.sup.B, and b1 are as defined
herein.
[0331] In certain embodiments, the compound of Formula (II') is of
the formula:
##STR00196##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof.
[0332] In certain embodiments, a compound of Formula (II) is of
Formula (II-ii):
##STR00197##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof; wherein L.sup.2, X, Ring A,
R.sup.1, R.sup.2, R.sup.A, R.sup.B, a1, and b1 are as defined
herein.
[0333] In certain embodiments, a compound of Formula (II) is of the
formula:
##STR00198##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof.
[0334] In certain embodiments, a compound of Formula (II) is of the
formula:
##STR00199##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof.
[0335] In certain embodiments, a compound of Formula (II) is of
Formula (II-iii):
##STR00200##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof wherein L.sup.2, X, Ring A, R.sup.1,
R.sup.2, R.sup.A, R.sup.B, a1, and b1 are as defined herein.
[0336] In certain embodiments, a compound of Formula (II) is of the
formula:
##STR00201##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof.
[0337] In certain embodiments, a compound of Formula (II) is of the
formula:
##STR00202##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof.
[0338] In certain embodiments, a compound of Formula (I') is of the
formula:
##STR00203## ##STR00204## ##STR00205## ##STR00206## ##STR00207##
##STR00208## ##STR00209## ##STR00210##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof.
[0339] In certain embodiments, a compound of Formula (I) is of the
formula:
##STR00211## ##STR00212## ##STR00213## ##STR00214##
##STR00215##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof.
[0340] In certain embodiments, a compound of Formula (I) is of the
formula:
##STR00216## ##STR00217## ##STR00218## ##STR00219## ##STR00220##
##STR00221## ##STR00222## ##STR00223## ##STR00224## ##STR00225##
##STR00226## ##STR00227## ##STR00228## ##STR00229## ##STR00230##
##STR00231## ##STR00232## ##STR00233## ##STR00234## ##STR00235##
##STR00236##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof.
[0341] In certain embodiments, a compound of Formula (II') is of
the formula:
##STR00237##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof.
[0342] In certain embodiments, a compound of Formula (II) is of the
formula:
##STR00238## ##STR00239##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof.
Pharmaceutical Compositions and Administration
[0343] The pharmaceutical compositions described herein may be
useful in treating and/or preventing proliferative diseases (e.g.,
cancers (e.g., leukemia, acute lymphoblastic leukemia, lymphoma,
Burkitt's lymphoma, melanoma, multiple myeloma, breast cancer,
Ewing's sarcoma, osteosarcoma, brain cancer, neuroblastoma, lung
cancer, colorectal cancer), benign neoplasms, diseases associated
with angiogenesis, inflammatory diseases, autoinflammatory
diseases, and autoimmune diseases) in a subject. The compositions
described herein may also be useful for inhibiting the activity of
a protein kinase (e.g., CDK (e.g., CDK7, CDK12, and/or CDK13)) in a
subject, biological sample, tissue, or cell. The compositions
described herein may also be useful for inducing apoptosis in a
cell.
[0344] The present disclosure provides pharmaceutical compositions
comprising a compound described herein (e.g., a compound of any one
of Formulae (I'), (II'), (I), (II), or a pharmaceutically
acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer,
stereoisomer, isotopically labeled derivative, or prodrug thereof,
and optionally a pharmaceutically acceptable excipient. In certain
embodiments, the pharmaceutical composition of the invention
comprises a compound described herein, or a pharmaceutically
acceptable salt thereof, and optionally a pharmaceutically
acceptable excipient. In certain embodiments, a pharmaceutical
composition described herein comprises a compound described herein,
or a pharmaceutically acceptable salt thereof, and a
pharmaceutically acceptable excipient. In certain embodiments, the
compound described herein, or a pharmaceutically acceptable salt,
solvate, hydrate, polymorph, co-crystal, tautomer, stereoisomer,
isotopically labeled derivative, or prodrug thereof, is provided in
an effective amount in the pharmaceutical composition.
[0345] In certain embodiments, the effective amount is a
therapeutically effective amount (e.g., amount effective for
treating a proliferative disease in a subject in need thereof). In
certain embodiments, the effective amount is an amount effective
for inhibiting the activity of a protein kinase (e.g., CDK (e.g.,
CDK7, CDK12, and/or CDK13)) in a subject in need thereof. In
certain embodiments, the effective amount is an amount effective
for inhibiting the activity of a protein kinase (e.g., CDK (e.g.,
CDK7, CDK12, and/or CDK13)) in a cell. In certain embodiments, the
effective amount is an amount effective for inducing apoptosis in a
cell. In certain embodiments, the effective amount is a
prophylactically effective amount (e.g., amount effective for
preventing a proliferative disease in a subject in need thereof
and/or for keeping a subject in need thereof in remission of a
proliferative disease).
[0346] In certain embodiments, the protein kinase being inhibited
is a CDK. In certain embodiments, the protein kinase being
inhibited is CDK1, CDK2, CDK3, CDK4, CDK5, CDK6, CDK7, CDK8, CDK9,
CDK10, CDK11, CDK12, CDK13, CDK14, CDK15, CDK16, CDK17, CDK18,
CDK19, or CDK20. In certain embodiments, the protein kinase being
inhibited is CDK7. In certain embodiments, the protein kinase being
inhibited is CDK12. In certain embodiments, the protein kinase
being inhibited is CDK13. In certain embodiments, the protein
kinase being inhibited is a Src family kinase. In certain
embodiments, the protein kinase being inhibited is SRC. In certain
embodiments, the protein kinase being inhibited is FGR. In certain
embodiments, the protein kinase being inhibited is BUB1B. In
certain embodiments, the protein kinase being inhibited is CHEK2.
In certain embodiments, the protein kinase being inhibited is
HIPK4. In certain embodiments, the protein kinase being inhibited
is PRKCQ. In certain embodiments, the protein kinase being
inhibited is R.sup.EET. In certain embodiments, the protein kinase
being inhibited is MELK. In certain embodiments, the protein kinase
being inhibited is IRAK1, IRAK4, BMX, or PI3K. In certain
embodiments, the protein kinase being inhibited is ABL, ARG, BLK,
CSK, EphB1, EphB2, FGR, FRK, FYN, SRC, YES, LCK, LYN, MAP2K5, NLK,
p38a, SNRK, or TEC. In certain embodiments, the protein kinase
being inhibited is ABL1(H396P)-phosphorylated, ABL1-phosphorylated,
BLK, EPHA4, EPHB2, EPHB3, EPHB4, FGR, JAK3(JH1domain-catalytic),
KIT, KIT(L576P), KIT(V559D), PDGFRB, SRC, YES,
ABL1(H396P)-nonphosphorylated, ABL1 (Y253F)-phosphorylated,
ABL1-nonphosphorylated, FRK, LYN, ABL1(Q252H)-nonphosphorylated,
DDR1, EPHB1, ERBB4, p38-alpha, ABL2, ABL1(Q252H)-phosphorylated,
SIK, EPHA8, MEK5, ABL1(E255K)-phosphorylated,
ABL1(F317L)-nonphosphorylated, FYN, LCK, EPHA2,
ABL1(M351T)-phosphorylated, TXK, EGFR(L858R), EGFR(L861Q), ERBB2,
ERBB3, EPHA5, ABL1(F317I)-nonphosphorylated, EGFR(L747-E749del,
A750P), CSK, EPHA1, ABL1(F317L)-phosphorylated, BRAF(V600E), EGFR,
KIT-autoinhibited, or EGFR(E746-A750del). In certain embodiments,
the protein kinase being inhibited is
ABL1(F317L)-nonphosphorylated, ABL1 (H396P)-nonphosphorylated, ABL1
(H396P)-phosphorylated, ABL1-phosphorylated, BLK, EPHA4, EPHB2,
EPHB3, EPHB4, JAK3(JH1domain-catalytic), KIT, KIT(L576P),
KIT(V559D), LYN, PDGFRB, SRC, YES, ABL1-nonphosphorylated,
ABL1(Y253F)-phosphorylated, ERBB3, FGR, FRK, p38-alpha,
ABL1(F317I)-nonphosphorylated, DDR1, EPHA2, ABL1
(Q252H)-phosphorylated, MEK5, ABL1 (Q252H)-nonphosphorylated, ABL2,
FYN, EPHB1, ABL1(E255K)-phosphorylated, ABL1
(F317L)-phosphorylated, EPHA1, ABL1(M351T)-phosphorylated, ERBB4,
TXK, LCK, EPHA8, SIK, EPHA5, EGFR(L861Q), CSF1R-autoinhibited,
BRAF(V600E), BRK, CSK, KIT(D816V), KIT-autoinhibited,
EGFR(L747-T751del,Sins), EGFR(L858R), EGFR(L747-E749del, A750P), or
CSF1R.
[0347] In certain embodiments, the effective amount is an amount
effective for inhibiting the activity of a protein kinase (e.g.,
CDK (e.g., CDK7, CDK12, and/or CDK13)) by at least about 10%, at
least about 20%, at least about 30%, at least about 40%, at least
about 50%, at least about 60%, at least about 70%, at least about
80%, at least about 90%, at least about 95%, or at least about 98%.
In certain embodiments, the effective amount is an amount effective
for inhibiting the activity of a protein kinase (e.g., CDK (e.g.,
CDK7, CDK12, and/or CDK13)) by not more than 10%, not more than
20%, not more than 30%, not more than 40%, not more than 50%, not
more than 60%, not more than 70%, not more than 80%, not more than
90%, not more than 95%, or not more than 98%. In certain
embodiments, the effective amount is an amount effective for
inhibiting the activity of a protein kinase (e.g., CDK (e.g., CDK7,
CDK12, and/or CDK13)) by a range between a percentage described in
this paragraph and another percentage described in this paragraph,
inclusive.
[0348] Pharmaceutical compositions described herein can be prepared
by any method known in the art of pharmacology. In general, such
preparatory methods include bringing the compound described herein
(i.e., the "active ingredient") into association with a carrier or
excipient, and/or one or more other accessory ingredients, and
then, if necessary and/or desirable, shaping, and/or packaging the
product into a desired single- or multi-dose unit.
[0349] Pharmaceutical compositions can be prepared, packaged,
and/or sold in bulk, as a single unit dose, and/or as a plurality
of single unit doses. A "unit dose" is a discrete amount of the
pharmaceutical composition comprising a predetermined amount of the
active ingredient. The amount of the active ingredient is generally
equal to the dosage of the active ingredient which would be
administered to a subject and/or a convenient fraction of such a
dosage, such as one-half or one-third of such a dosage.
[0350] Relative amounts of the active ingredient, the
pharmaceutically acceptable excipient, and/or any additional
ingredients in a pharmaceutical composition described herein will
vary, depending upon the identity, size, and/or condition of the
subject treated and further depending upon the route by which the
composition is to be administered. The composition may comprise
between about 0.1% and about 100% (w/w) of the active
ingredient.
[0351] Pharmaceutically acceptable excipients used in the
manufacture of provided pharmaceutical compositions include inert
diluents, dispersing and/or granulating agents, surface active
agents and/or emulsifiers, disintegrating agents, binding agents,
preservatives, buffering agents, lubricating agents, and/or oils.
Excipients such as cocoa butter and suppository waxes, coloring
agents, coating agents, sweetening, flavoring, and perfuming agents
may also be present in the composition.
[0352] Exemplary diluents include calcium carbonate, sodium
carbonate, calcium phosphate, dicalcium phosphate, calcium sulfate,
calcium hydrogen phosphate, sodium phosphate lactose, sucrose,
cellulose, microcrystalline cellulose, kaolin, mannitol, sorbitol,
inositol, sodium chloride, dry starch, cornstarch, powdered sugar,
and mixtures thereof.
[0353] Exemplary granulating and/or dispersing agents include
potato starch, corn starch, tapioca starch, sodium starch
glycolate, clays, alginic acid, guar gum, citrus pulp, agar,
bentonite, cellulose, and wood products, natural sponge,
cation-exchange resins, calcium carbonate, silicates, sodium
carbonate, cross-linked poly(vinyl-pyrrolidone) (crospovidone),
sodium carboxymethyl starch (sodium starch glycolate),
carboxymethyl cellulose, cross-linked sodium carboxymethyl
cellulose (croscarmellose), methylcellulose, pregelatinized starch
(starch 1500), microcrystalline starch, water insoluble starch,
calcium carboxymethyl cellulose, magnesium aluminum silicate
(Veegum), sodium lauryl sulfate, quaternary ammonium compounds, and
mixtures thereof.
[0354] Exemplary surface active agents and/or emulsifiers include
natural emulsifiers (e.g., acacia, agar, alginic acid, sodium
alginate, tragacanth, chondrux, cholesterol, xanthan, pectin,
gelatin, egg yolk, casein, wool fat, cholesterol, wax, and
lecithin), colloidal clays (e.g., bentonite (aluminum silicate) and
Veegum (magnesium aluminum silicate)), long chain amino acid
derivatives, high molecular weight alcohols (e.g., stearyl alcohol,
cetyl alcohol, oleyl alcohol, triacetin monostearate, ethylene
glycol distearate, glyceryl monostearate, and propylene glycol
monostearate, polyvinyl alcohol), carbomers (e.g., carboxy
polymethylene, polyacrylic acid, acrylic acid polymer, and
carboxyvinyl polymer), carrageenan, cellulosic derivatives (e.g.,
carboxymethylcellulose sodium, powdered cellulose, hydroxymethyl
cellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose,
methylcellulose), sorbitan fatty acid esters (e.g., polyoxyethylene
sorbitan monolaurate (Tween.RTM. 20), polyoxyethylene sorbitan
(Tween.RTM. 60), polyoxyethylene sorbitan monooleate (Tween.RTM.
80), sorbitan monopalmitate (Span.RTM. 40), sorbitan monostearate
(Span.RTM. 60), sorbitan tristearate (Span.RTM.65), glyceryl
monooleate, sorbitan monooleate (Spana 80), polyoxyethylene esters
(e.g., polyoxyethylene monostearate (Myrj.RTM.45), polyoxyethylene
hydrogenated castor oil, polyethoxylated castor oil,
polyoxymethylene stearate, and Solutol.RTM.), sucrose fatty acid
esters, polyethylene glycol fatty acid esters (e.g.,
Cremophor.RTM.), polyoxyethylene ethers, (e.g., polyoxyethylene
lauryl ether (Brij.RTM. 30)), poly(vinyl-pyrrolidone), diethylene
glycol monolaurate, triethanolamine oleate, sodium oleate,
potassium oleate, ethyl oleate, oleic acid, ethyl laurate, sodium
lauryl sulfate, Pluronic.RTM. F-68, poloxamer P-188, cetrimonium
bromide, cetylpyridinium chloride, benzalkonium chloride, docusate
sodium, and/or mixtures thereof.
[0355] Exemplary binding agents include starch (e.g., cornstarch
and starch paste), gelatin, sugars (e.g., sucrose, glucose,
dextrose, dextrin, molasses, lactose, lactitol, mannitol, etc.),
natural and synthetic gums (e.g., acacia, sodium alginate, extract
of Irish moss, panwar gum, ghatti gum, mucilage of isapol husks,
carboxymethylcellulose, methylcellulose, ethylcellulose,
hydroxyethylcellulose, hydroxypropyl cellulose, hydroxypropyl
methylcellulose, microcrystalline cellulose, cellulose acetate,
poly(vinyl-pyrrolidone), magnesium aluminum silicate (Veegum.RTM.),
and larch arabogalactan), alginates, polyethylene oxide,
polyethylene glycol, inorganic calcium salts, silicic acid,
polymethacrylates, waxes, water, alcohol, and/or mixtures
thereof.
[0356] Exemplary preservatives include antioxidants, chelating
agents, antimicrobial preservatives, antifungal preservatives,
antiprotozoan preservatives, alcohol preservatives, acidic
preservatives, and other preservatives. In certain embodiments, the
preservative is an antioxidant. In other embodiments, the
preservative is a chelating agent.
[0357] Exemplary antioxidants include alpha tocopherol, ascorbic
acid, ascorbyl palmitate, butylated hydroxyanisole, butylated
hydroxytoluene, monothioglycerol, potassium metabisulfite,
propionic acid, propyl gallate, sodium ascorbate, sodium bisulfite,
sodium metabisulfite, and sodium sulfite.
[0358] Exemplary chelating agents include
ethylenediaminetetraacetic acid (EDTA) and salts and hydrates
thereof (e.g., sodium edetate, disodium edetate, trisodium edetate,
calcium disodium edetate, dipotassium edetate, and the like),
citric acid and salts and hydrates thereof (e.g., citric acid
monohydrate), fumaric acid and salts and hydrates thereof, malic
acid and salts and hydrates thereof, phosphoric acid and salts and
hydrates thereof, and tartaric acid and salts and hydrates thereof.
Exemplary antimicrobial preservatives include benzalkonium
chloride, benzethonium chloride, benzyl alcohol, bronopol,
cetrimide, cetylpyridinium chloride, chlorhexidine, chlorobutanol,
chlorocresol, chloroxylenol, cresol, ethyl alcohol, glycerin,
hexetidine, imidurea, phenol, phenoxyethanol, phenylethyl alcohol,
phenylmercuric nitrate, propylene glycol, and thimerosal.
[0359] Exemplary antifungal preservatives include butyl paraben,
methyl paraben, ethyl paraben, propyl paraben, benzoic acid,
hydroxybenzoic acid, potassium benzoate, potassium sorbate, sodium
benzoate, sodium propionate, and sorbic acid.
[0360] Exemplary alcohol preservatives include ethanol,
polyethylene glycol, phenol, phenolic compounds, bisphenol,
chlorobutanol, hydroxybenzoate, and phenylethyl alcohol.
[0361] Exemplary acidic preservatives include vitamin A, vitamin C,
vitamin E, beta-carotene, citric acid, acetic acid, dehydroacetic
acid, ascorbic acid, sorbic acid, and phytic acid.
[0362] Other preservatives include tocopherol, tocopherol acetate,
deteroxime mesylate, cetrimide, butylated hydroxyanisol (BHA),
butylated hydroxytoluened (BHT), ethylenediamine, sodium lauryl
sulfate (SLS), sodium lauryl ether sulfate (SLES), sodium
bisulfite, sodium metabisulfite, potassium sulfite, potassium
metabisulfite, Glydant.RTM. Plus, Phenonip.RTM., methylparaben,
Germall.RTM. 115, Germaben.RTM. II, Neolone.RTM., Kathon.RTM., and
Euxyl.RTM..
[0363] Exemplary buffering agents include citrate buffer solutions,
acetate buffer solutions, phosphate buffer solutions, ammonium
chloride, calcium carbonate, calcium chloride, calcium citrate,
calcium glubionate, calcium gluceptate, calcium gluconate,
D-gluconic acid, calcium glycerophosphate, calcium lactate,
propanoic acid, calcium levulinate, pentanoic acid, dibasic calcium
phosphate, phosphoric acid, tribasic calcium phosphate, calcium
hydroxide phosphate, potassium acetate, potassium chloride,
potassium gluconate, potassium mixtures, dibasic potassium
phosphate, monobasic potassium phosphate, potassium phosphate
mixtures, sodium acetate, sodium bicarbonate, sodium chloride,
sodium citrate, sodium lactate, dibasic sodium phosphate, monobasic
sodium phosphate, sodium phosphate mixtures, tromethamine,
magnesium hydroxide, aluminum hydroxide, alginic acid, pyrogen-free
water, isotonic saline, Ringer's solution, ethyl alcohol, and
mixtures thereof.
[0364] Exemplary lubricating agents include magnesium stearate,
calcium stearate, stearic acid, silica, talc, malt, glyceryl
behanate, hydrogenated vegetable oils, polyethylene glycol, sodium
benzoate, sodium acetate, sodium chloride, leucine, magnesium
lauryl sulfate, sodium lauryl sulfate, and mixtures thereof.
[0365] Exemplary natural oils include almond, apricot kernel,
avocado, babassu, bergamot, black current seed, borage, cade,
camomile, canola, caraway, carnauba, castor, cinnamon, cocoa
butter, coconut, cod liver, coffee, corn, cotton seed, emu,
eucalyptus, evening primrose, fish, flaxseed, geraniol, gourd,
grape seed, hazel nut, hyssop, isopropyl myristate, jojoba, kukui
nut, lavandin, lavender, lemon, litsea cubeba, macademia nut,
mallow, mango seed, meadowfoam seed, mink, nutmeg, olive, orange,
orange roughy, palm, palm kernel, peach kernel, peanut, poppy seed,
pumpkin seed, rapeseed, rice bran, rosemary, safflower, sandalwood,
sasquana, savoury, sea buckthorn, sesame, shea butter, silicone,
soybean, sunflower, tea tree, thistle, tsubaki, vetiver, walnut,
and wheat germ oils. Exemplary synthetic oils include, but are not
limited to, butyl stearate, caprylic triglyceride, capric
triglyceride, cyclomethicone, diethyl sebacate, dimethicone 360,
isopropyl myristate, mineral oil, octyldodecanol, oleyl alcohol,
silicone oil, and mixtures thereof.
[0366] Liquid dosage forms for oral and parenteral administration
include pharmaceutically acceptable emulsions, microemulsions,
solutions, suspensions, syrups and elixirs. In addition to the
active ingredients, the liquid dosage forms may comprise inert
diluents commonly used in the art such as, for example, water or
other solvents, solubilizing agents and emulsifiers such as ethyl
alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl
alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol,
dimethylformamide, oils (e.g., cottonseed, groundnut, corn, germ,
olive, castor, and sesame oils), glycerol, tetrahydrofurfuryl
alcohol, polyethylene glycols and fatty acid esters of sorbitan,
and mixtures thereof. Besides inert diluents, the oral compositions
can include adjuvants such as wetting agents, emulsifying and
suspending agents, sweetening, flavoring, and perfuming agents. In
certain embodiments for parenteral administration, the conjugates
described herein are mixed with solubilizing agents such as
Cremophor.RTM., alcohols, oils, modified oils, glycols,
polysorbates, cyclodextrins, polymers, and mixtures thereof.
[0367] Injectable preparations, for example, sterile injectable
aqueous or oleaginous suspensions can be formulated according to
the known art using suitable dispersing or wetting agents and
suspending agents. The sterile injectable preparation can be a
sterile injectable solution, suspension, or emulsion in a nontoxic
parenterally acceptable diluent or solvent, for example, as a
solution in 1,3-butanediol. Among the acceptable vehicles and
solvents that can be employed are water, Ringer's solution, U.S.P.,
and isotonic sodium chloride solution. In addition, sterile, fixed
oils are conventionally employed as a solvent or suspending medium.
For this purpose any bland fixed oil can be employed including
synthetic mono- or di-glycerides. In addition, fatty acids such as
oleic acid are used in the preparation of injectables.
[0368] The injectable formulations can be sterilized, for example,
by filtration through a bacterial-retaining filter, or by
incorporating sterilizing agents in the form of sterile solid
compositions which can be dissolved or dispersed in sterile water
or other sterile injectable medium prior to use.
[0369] In order to prolong the effect of a drug, it is often
desirable to slow the absorption of the drug from subcutaneous or
intramuscular injection. This can be accomplished by the use of a
liquid suspension of crystalline or amorphous material with poor
water solubility. The rate of absorption of the drug then depends
upon its rate of dissolution, which, in turn, may depend upon
crystal size and crystalline form. Alternatively, delayed
absorption of a parenterally administered drug form may be
accomplished by dissolving or suspending the drug in an oil
vehicle.
[0370] Compositions for rectal or vaginal administration are
typically suppositories which can be prepared by mixing the
conjugates described herein with suitable non-irritating excipients
or carriers such as cocoa butter, polyethylene glycol, or a
suppository wax which are solid at ambient temperature but liquid
at body temperature and therefore melt in the rectum or vaginal
cavity and release the active ingredient.
[0371] Solid dosage forms for oral administration include capsules,
tablets, pills, powders, and granules. In such solid dosage forms,
the active ingredient is mixed with at least one inert,
pharmaceutically acceptable excipient or carrier such as sodium
citrate or dicalcium phosphate and/or (a) fillers or extenders such
as starches, lactose, sucrose, glucose, mannitol, and silicic acid,
(b) binders such as, for example, carboxymethylcellulose,
alginates, gelatin, polyvinylpyrrolidinone, sucrose, and acacia,
(c) humectants such as glycerol, (d) disintegrating agents such as
agar, calcium carbonate, potato or tapioca starch, alginic acid,
certain silicates, and sodium carbonate, (e) solution retarding
agents such as paraffin, (f) absorption accelerators such as
quaternary ammonium compounds, (g) wetting agents such as, for
example, cetyl alcohol and glycerol monostearate, (h) absorbents
such as kaolin and bentonite clay, and (i) lubricants such as talc,
calcium stearate, magnesium stearate, solid polyethylene glycols,
sodium lauryl sulfate, and mixtures thereof. In the case of
capsules, tablets, and pills, the dosage form may include a
buffering agent.
[0372] Solid compositions of a similar type can be employed as
fillers in soft and hard-filled gelatin capsules using such
excipients as lactose or milk sugar as well as high molecular
weight polyethylene glycols and the like. The solid dosage forms of
tablets, dragees, capsules, pills, and granules can be prepared
with coatings and shells such as enteric coatings and other
coatings well known in the art of pharmacology. They may optionally
comprise opacifying agents and can be of a composition that they
release the active ingredient(s) only, or preferentially, in a
certain part of the intestinal tract, optionally, in a delayed
manner. Examples of encapsulating compositions which can be used
include polymeric substances and waxes. Solid compositions of a
similar type can be employed as fillers in soft and hard-filled
gelatin capsules using such excipients as lactose or milk sugar as
well as high molecular weight polyethylene glycols and the
like.
[0373] The active ingredient can be in a micro-encapsulated form
with one or more excipients as noted above. The solid dosage forms
of tablets, dragees, capsules, pills, and granules can be prepared
with coatings and shells such as enteric coatings, release
controlling coatings, and other coatings well known in the
pharmaceutical formulating art. In such solid dosage forms the
active ingredient can be admixed with at least one inert diluent
such as sucrose, lactose, or starch. Such dosage forms may
comprise, as is normal practice, additional substances other than
inert diluents, e.g., tableting lubricants and other tableting aids
such a magnesium stearate and microcrystalline cellulose. In the
case of capsules, tablets and pills, the dosage forms may comprise
buffering agents. They may optionally comprise opacifying agents
and can be of a composition that they release the active
ingredient(s) only, or preferentially, in a certain part of the
intestinal tract, optionally, in a delayed manner. Examples of
encapsulating agents which can be used include polymeric substances
and waxes.
[0374] Dosage forms for topical and/or transdermal administration
of a compound described herein may include ointments, pastes,
creams, lotions, gels, powders, solutions, sprays, inhalants,
and/or patches. Generally, the active ingredient is admixed under
sterile conditions with a pharmaceutically acceptable carrier or
excipient and/or any needed preservatives and/or buffers as can be
required. Additionally, the present disclosure contemplates the use
of transdermal patches, which often have the added advantage of
providing controlled delivery of an active ingredient to the body.
Such dosage forms can be prepared, for example, by dissolving
and/or dispensing the active ingredient in the proper medium.
Alternatively or additionally, the rate can be controlled by either
providing a rate controlling membrane and/or by dispersing the
active ingredient in a polymer matrix and/or gel.
[0375] Suitable devices for use in delivering intradermal
pharmaceutical compositions described herein include short needle
devices. Intradermal compositions can be administered by devices
which limit the effective penetration length of a needle into the
skin. Alternatively or additionally, conventional syringes can be
used in the classical mantoux method of intradermal administration.
Jet injection devices which deliver liquid formulations to the
dermis via a liquid jet injector and/or via a needle which pierces
the stratum corneum and produces a jet which reaches the dermis are
suitable. Ballistic powder/particle delivery devices which use
compressed gas to accelerate the compound in powder form through
the outer layers of the skin to the dermis are suitable.
[0376] Formulations suitable for topical administration include,
but are not limited to, liquid and/or semi-liquid preparations such
as liniments, lotions, oil-in-water and/or water-in-oil emulsions
such as creams, ointments, and/or pastes, and/or solutions and/or
suspensions. Topically administrable formulations may, for example,
comprise from about 1% to about 10% (w/w) active ingredient,
although the concentration of the active ingredient can be as high
as the solubility limit of the active ingredient in the solvent.
Formulations for topical administration may further comprise one or
more of the additional ingredients described herein.
[0377] A pharmaceutical composition described herein can be
prepared, packaged, and/or sold in a formulation suitable for
pulmonary administration via the buccal cavity. Such a formulation
may comprise dry particles which comprise the active ingredient and
which have a diameter in the range from about 0.5 to about 7
nanometers, or from about 1 to about 6 nanometers. Such
compositions are conveniently in the form of dry powders for
administration using a device comprising a dry powder reservoir to
which a stream of propellant can be directed to disperse the powder
and/or using a self-propelling solvent/powder dispensing container
such as a device comprising the active ingredient dissolved and/or
suspended in a low-boiling propellant in a sealed container. Such
powders comprise particles wherein at least 98% of the particles by
weight have a diameter greater than 0.5 nanometers and at least 95%
of the particles by number have a diameter less than 7 nanometers.
Alternatively, at least 95% of the particles by weight have a
diameter greater than 1 nanometer and at least 90% of the particles
by number have a diameter less than 6 nanometers. Dry powder
compositions may include a solid fine powder diluent such as sugar
and are conveniently provided in a unit dose form.
[0378] Low boiling propellants generally include liquid propellants
having a boiling point of below 65.degree. F. at atmospheric
pressure. Generally the propellant may constitute 50 to 99.9% (w/w)
of the composition, and the active ingredient may constitute 0.1 to
20% (w/w) of the composition. The propellant may further comprise
additional ingredients such as a liquid non-ionic and/or solid
anionic surfactant and/or a solid diluent (which may have a
particle size of the same order as particles comprising the active
ingredient).
[0379] Pharmaceutical compositions described herein formulated for
pulmonary delivery may provide the active ingredient in the form of
droplets of a solution and/or suspension. Such formulations can be
prepared, packaged, and/or sold as aqueous and/or dilute alcoholic
solutions and/or suspensions, optionally sterile, comprising the
active ingredient, and may conveniently be administered using any
nebulization and/or atomization device. Such formulations may
further comprise one or more additional ingredients including, but
not limited to, a flavoring agent such as saccharin sodium, a
volatile oil, a buffering agent, a surface active agent, and/or a
preservative such as methylhydroxybenzoate. The droplets provided
by this route of administration may have an average diameter in the
range from about 0.1 to about 200 nanometers.
[0380] Formulations described herein as being useful for pulmonary
delivery are useful for intranasal delivery of a pharmaceutical
composition described herein. Another formulation suitable for
intranasal administration is a coarse powder comprising the active
ingredient and having an average particle from about 0.2 to 500
micrometers. Such a formulation is administered by rapid inhalation
through the nasal passage from a container of the powder held close
to the nares.
[0381] Formulations for nasal administration may, for example,
comprise from about as little as 0.1% (w/w) to as much as 100%
(w/w) of the active ingredient, and may comprise one or more of the
additional ingredients described herein. A pharmaceutical
composition described herein can be prepared, packaged, and/or sold
in a formulation for buccal administration. Such formulations may,
for example, be in the form of tablets and/or lozenges made using
conventional methods, and may contain, for example, 0.1 to 20%
(w/w) active ingredient, the balance comprising an orally
dissolvable and/or degradable composition and, optionally, one or
more of the additional ingredients described herein. Alternately,
formulations for buccal administration may comprise a powder and/or
an aerosolized and/or atomized solution and/or suspension
comprising the active ingredient. Such powdered, aerosolized,
and/or aerosolized formulations, when dispersed, may have an
average particle and/or droplet size in the range from about 0.1 to
about 200 nanometers, and may further comprise one or more of the
additional ingredients described herein.
[0382] A pharmaceutical composition described herein can be
prepared, packaged, and/or sold in a formulation for ophthalmic
administration. Such formulations may, for example, be in the form
of eye drops including, for example, a 0.1-1.0% (w/w) solution
and/or suspension of the active ingredient in an aqueous or oily
liquid carrier or excipient. Such drops may further comprise
buffering agents, salts, and/or one or more other of the additional
ingredients described herein. Other opthalmically-administrable
formulations which are useful include those which comprise the
active ingredient in microcrystalline form and/or in a liposomal
preparation. Ear drops and/or eye drops are also contemplated as
being within the scope of this disclosure.
[0383] Although the descriptions of pharmaceutical compositions
provided herein are principally directed to pharmaceutical
compositions which are suitable for administration to humans, such
compositions are generally suitable for administration to animals
of all sorts. Modification of pharmaceutical compositions suitable
for administration to humans in order to render the compositions
suitable for administration to various animals is well understood,
and the ordinarily skilled veterinary pharmacologist can design
and/or perform such modification with ordinary experimentation.
[0384] The compounds provided herein are typically formulated in
dosage unit form for ease of administration and uniformity of
dosage. It will be understood, however, that the total daily usage
of the compositions described herein will be decided by a physician
within the scope of sound medical judgment. The specific
therapeutically effective dose level for any particular subject or
organism will depend upon a variety of factors including the
disease being treated and the severity of the disorder; the
activity of the specific active ingredient employed; the specific
composition employed; the age, body weight, general health, sex,
and diet of the subject; the time of administration, route of
administration, and rate of excretion of the specific active
ingredient employed; the duration of the treatment; drugs used in
combination or coincidental with the specific active ingredient
employed; and like factors well known in the medical arts.
[0385] The compounds and compositions provided herein can be
administered by any route, including enteral (e.g., oral),
parenteral, intravenous, intramuscular, intra-arterial,
intramedullary, intrathecal, subcutaneous, intraventricular,
transdermal, interdermal, rectal, intravaginal, intraperitoneal,
topical (as by powders, ointments, creams, and/or drops), mucosal,
nasal, bucal, sublingual; by intratracheal instillation, bronchial
instillation, and/or inhalation; and/or as an oral spray, nasal
spray, and/or aerosol. Specifically contemplated routes are oral
administration, intravenous administration (e.g., systemic
intravenous injection), regional administration via blood and/or
lymph supply, and/or direct administration to an affected site. In
general, the most appropriate route of administration will depend
upon a variety of factors including the nature of the agent (e.g.,
its stability in the environment of the gastrointestinal tract),
and/or the condition of the subject (e.g., whether the subject is
able to tolerate oral administration). In certain embodiments, the
compound or pharmaceutical composition described herein is suitable
for topical administration to the eye of a subject.
[0386] The exact amount of a compound required to achieve an
effective amount will vary from subject to subject, depending, for
example, on species, age, and general condition of a subject,
severity of the side effects or disorder, identity of the
particular compound, mode of administration, and the like. An
effective amount may be included in a single dose (e.g., single
oral dose) or multiple doses (e.g., multiple oral doses). In
certain embodiments, when multiple doses are administered to a
subject or applied to a biological sample, tissue, or cell, any two
doses of the multiple doses include different or substantially the
same amounts of a compound described herein. In certain
embodiments, when multiple doses are administered to a subject or
applied to a biological sample, tissue, or cell, the frequency of
administering the multiple doses to the subject or applying the
multiple doses to the tissue or cell is three doses a day, two
doses a day, one dose a day, one dose every other day, one dose
every third day, one dose every week, one dose every two weeks, one
dose every three weeks, or one dose every four weeks. In certain
embodiments, the frequency of administering the multiple doses to
the subject or applying the multiple doses to the tissue or cell is
one dose per day. In certain embodiments, the frequency of
administering the multiple doses to the subject or applying the
multiple doses to the tissue or cell is two doses per day. In
certain embodiments, the frequency of administering the multiple
doses to the subject or applying the multiple doses to the tissue
or cell is three doses per day. In certain embodiments, when
multiple doses are administered to a subject or applied to a
biological sample, tissue, or cell, the duration between the first
dose and last dose of the multiple doses is one day, two days, four
days, one week, two weeks, three weeks, one month, two months,
three months, four months, six months, nine months, one year, two
years, three years, four years, five years, seven years, ten years,
fifteen years, twenty years, or the lifetime of the subject,
biological sample, tissue, or cell. In certain embodiments, the
duration between the first dose and last dose of the multiple doses
is three months, six months, or one year. In certain embodiments,
the duration between the first dose and last dose of the multiple
doses is the lifetime of the subject, biological sample, tissue, or
cell. In certain embodiments, a dose (e.g., a single dose, or any
dose of multiple doses) described herein includes independently
between 0.1 .mu.g and 1 .mu.g, between 0.001 mg and 0.01 mg,
between 0.01 mg and 0.1 mg, between 0.1 mg and 1 mg, between 1 mg
and 3 mg, between 3 mg and 10 mg, between 10 mg and 30 mg, between
30 mg and 100 mg, between 100 mg and 300 mg, between 300 mg and
1,000 mg, or between 1 g and 10 g, inclusive, of a compound
described herein. In certain embodiments, a dose described herein
includes independently between 1 mg and 3 mg, inclusive, of a
compound described herein. In certain embodiments, a dose described
herein includes independently between 3 mg and 10 mg, inclusive, of
a compound described herein. In certain embodiments, a dose
described herein includes independently between 10 mg and 30 mg,
inclusive, of a compound described herein. In certain embodiments,
a dose described herein includes independently between 30 mg and
100 mg, inclusive, of a compound described herein.
[0387] Dose ranges as described herein provide guidance for the
administration of provided pharmaceutical compositions to an adult.
The amount to be administered to, for example, a child or an
adolescent can be determined by a medical practitioner or person
skilled in the art and can be lower or the same as that
administered to an adult. In certain embodiments, a dose described
herein is a dose for an adult human whose body weight is
approximately 70 kg.
[0388] A compound or composition, as described herein, may be
administered in combination with one or more additional
pharmaceutical agents (e.g., therapeutically and/or
prophylactically active agents) useful in treating and/or
preventing a proliferative disease. The compounds or compositions
can be administered in combination with additional pharmaceutical
agents that improve their activity (e.g., activity (e.g., potency
and/or efficacy) in treating a proliferative disease in a subject
in need thereof, in preventing a proliferative disease in a subject
in need thereof, and/or in inhibiting the activity of a protein
kinase (e.g., CDK (e.g., CDK7, CDK12, and/or CDK13)) in a subject,
biological sample, tissue, or cell), improve bioavailability,
improve safety, reduce drug resistance, reduce and/or modify
metabolism, inhibit excretion, and/or modify distribution in a
subject, biological sample, tissue, or cell. It will also be
appreciated that the therapy employed may achieve a desired effect
for the same disorder, and/or it may achieve different effects. In
certain embodiments, a pharmaceutical composition described herein
including a compound described herein and an additional
pharmaceutical agent shows a synergistic effect that is absent in a
pharmaceutical composition including one of the compound and the
additional pharmaceutical agent, but not both.
[0389] The compound or composition can be administered concurrently
with, prior to, or subsequent to one or more additional
pharmaceutical agents, which are different from the compound or
composition and may be useful as, e.g., combination therapies in
treating and/or preventing a proliferative disease. Pharmaceutical
agents include therapeutically active agents. Pharmaceutical agents
also include prophylactically active agents. Pharmaceutical agents
include small organic molecules such as drug compounds (e.g.,
compounds approved for human or veterinary use by the U.S. Food and
Drug Administration as provided in the Code of Federal Regulations
(CFR)), peptides, proteins, carbohydrates, monosaccharides,
oligosaccharides, polysaccharides, nucleoproteins, mucoproteins,
lipoproteins, synthetic polypeptides or proteins, small molecules
linked to proteins, glycoproteins, steroids, nucleic acids, DNAs,
RNAs, nucleotides, nucleosides, oligonucleotides, antisense
oligonucleotides, lipids, hormones, vitamins, and cells. In certain
embodiments, the additional pharmaceutical agent is a
pharmaceutical agent useful in treating a proliferative disease. In
certain embodiments, the additional pharmaceutical agent is a
pharmaceutical agent useful in preventing a proliferative disease.
In certain embodiments, the additional pharmaceutical agent is a
pharmaceutical agent useful in inhibiting the activity of a protein
kinase (e.g., CDK (e.g., CDK7, CDK12, and/or CDK13)) in a subject,
biological sample, tissue, or cell. In certain embodiments, the
additional pharmaceutical agent is a pharmaceutical agent useful in
inducing apoptosis in a cell. In certain embodiments, the
additional pharmaceutical agent is a pharmaceutical agent approved
by a regulatory agency (e.g., the US FDA) for treating and/or
preventing a proliferative disease. Each additional pharmaceutical
agent may be administered at a dose and/or on a time schedule
determined for that pharmaceutical agent. The additional
pharmaceutical agent(s) may also be administered together with each
other and/or with the compound or composition described herein in a
single dose or administered separately in different doses. The
particular combination to employ in a regimen will take into
account compatibility of the compound described herein with the
additional pharmaceutical agent(s) and/or the desired therapeutic
and/or prophylactic effect to be achieved. In general, it is
expected that the additional pharmaceutical agent(s) in combination
be utilized at levels that do not exceed the levels at which they
are utilized individually. In some embodiments, the levels utilized
in combination will be lower than those utilized individually.
[0390] In certain embodiments, the additional pharmaceutical agent
is a cytotoxic agent. In certain embodiments, the additional
pharmaceutical agent is an anti-proliferative agent (e.g.,
anti-cancer agent). In certain embodiments, the additional
pharmaceutical agent is an anti-leukemia agent. In certain
embodiments, the additional pharmaceutical agent is ABITREXATE
(methotrexate), ADE, Adriamycin RDF (doxorubicin hydrochloride),
Ambochlorin (chlorambucil), ARRANON (nelarabine), ARZERRA
(ofatumumab), BOSULIF (bosutinib), BUSULFEX (busulfan), CAMPATH
(alemtuzumab), CERUBIDINE (daunorubicin hydrochloride), CLAFEN
(cyclophosphamide), CLOFAREX (clofarabine), CLOLAR (clofarabine),
CVP, CYTOSAR-U (cytarabine), CYTOXAN (cyclophosphamide), ERWINAZE
(Asparaginase Erwinia Chrysanthemi), FLUDARA (fludarabine
phosphate), FOLEX (methotrexate), FOLEX PFS (methotrexate), GAZYVA
(obinutuzumab), GLEEVEC (imatinib mesylate), Hyper-CVAD, ICLUSIG
(ponatinib hydrochloride), IMBRUVICA (ibrutinib), LEUKERAN
(chlorambucil), LINFOLIZIN (chlorambucil), MARQIBO (vincristine
sulfate liposome), METHOTREXATE LPF (methorexate), MEXATE
(methotrexate), MEXATE-AQ (methotrexate), mitoxantrone
hydrochloride, MUSTARGEN (mechlorethamine hydrochloride), MYLERAN
(busulfan), NEOSAR (cyclophosphamide), ONCASPAR (Pegaspargase),
PURINETHOL (mercaptopurine), PURIXAN (mercaptopurine), Rubidomycin
(daunorubicin hydrochloride), SPRYCEL (dasatinib), SYNRIBO
(omacetaxine mepesuccinate), TARABINE PFS (cytarabine), TASIGNA
(nilotinib), TREANDA (bendamustine hydrochloride), TRISENOX
(arsenic trioxide), VINCASAR PFS (vincristine sulfate), ZYDELIG
(idelalisib), or a combination thereof. In certain embodiments, the
additional pharmaceutical agent is an anti-lymphoma agent. In
certain embodiments, the additional pharmaceutical agent is
ABITREXATE (methotrexate), ABVD, ABVE, ABVE-PC, ADCETRIS
(brentuximab vedotin), ADRIAMYCIN PFS (doxorubicin hydrochloride),
ADRIAMYCIN RDF (doxorubicin hydrochloride), AMBOCHLORIN
(chlorambucil), AMBOCLORIN (chlorambucil), ARRANON (nelarabine),
BEACOPP, BECENUM (carmustine), BELEODAQ (belinostat), BEXXAR
(tositumomab and iodine I 131 tositumomab), BICNU (carmustine),
BLENOXANE (bleomycin), CARMUBRIS (carmustine), CHOP, CLAFEN
(cyclophosphamide), COPP, COPP-ABV, CVP, CYTOXAN
(cyclophosphamide), DEPOCYT (liposomal cytarabine), DTIC-DOME
(dacarbazine), EPOCH, FOLEX (methotrexate), FOLEX PFS
(methotrexate), FOLOTYN (pralatrexate), HYPER-CVAD, ICE, IMBRUVICA
(ibrutinib), INTRON A (recombinant interferon alfa-2b), ISTODAX
(romidepsin), LEUKERAN (chlorambucil), LINFOLIZIN (chlorambucil),
Lomustine, MATULANE (procarbazine hydrochloride), METHOTREXATE LPF
(methotrexate), MEXATE (methotrexate), MEXATE-AQ (methotrexate),
MOPP, MOZOBIL (plerixafor), MUSTARGEN (mechlorethamine
hydrochloride), NEOSAR (cyclophosphamide), OEPA, ONTAK (denileukin
diftitox), OPPA, R-CHOP, REVLIMID (lenalidomide), RITUXAN
(rituximab), STANFORD V, TREANDA (bendamustine hydrochloride),
VAMP, VELBAN (vinblastine sulfate), VELCADE (bortezomib), VELSAR
(vinblastine sulfate), VINCASAR PFS (vincristine sulfate), ZEVALIN
(ibritumomab tiuxetan), ZOLINZA (vorinostat), ZYDELIG (idelalisib),
or a combination thereof. In certain embodiments, the additional
pharmaceutical agent is ABITREXATE (methotrexate), ABRAXANE
(paclitaxel albumin-stabilized nanoparticle formulation), AC, AC-T,
ADE, ADRIAMYCIN PFS (doxorubicin hydrochloride), ADRUCIL
(fluorouracil), AFINITOR (everolimus), AFINITOR DISPERZ
(everolimus), ALDARA (imiquimod), ALIMTA (pemetrexed disodium),
AREDIA (pamidronate disodium), ARIMIDEX (anastrozole), AROMASIN
(exemestane), AVASTIN (bevacizumab), BECENUM (carmustine), BEP,
BICNU (carmustine), BLENOXANE (bleomycin), CAF, CAMPTOSAR
(irinotecan hydrochloride), CAPOX, CAPRELSA (vandetanib),
CARBOPLATIN-TAXOL, CARMUBRIS (carmustine), CASODEX (bicalutamide),
CEENU (lomustine), CERUBIDINE (daunorubicin hydrochloride),
CERVARIX (recombinant HPV bivalent vaccine), CLAFEN
(cyclophosphamide), CMF, COMETRIQ (cabozantinib-s-malate), COSMEGEN
(dactinomycin), CYFOS (ifosfamide), CYRAMZA (ramucirumab),
CYTOSAR-U (cytarabine), CYTOXAN (cyclophosphamide), DACOGEN
(decitabine), DEGARELIX, DOXIL (doxorubicin hydrochloride
liposome), DOXORUBICIN HYDROCHLORIDE, DOX-SL (doxorubicin
hydrochloride liposome), DTIC-DOME (dacarbazine), EFUDEX
(fluorouracil), ELLENCE (epirubicin hydrochloride), ELOXATIN
(oxaliplatin), ERBITUX (cetuximab), ERIVEDGE (vismodegib),
ETOPOPHOS (etoposide phosphate), EVACET (doxorubicin hydrochloride
liposome), FARESTON (toremifene), FASLODEX (fulvestrant), FEC,
FEMARA (letrozole), FLUOROPLEX (fluorouracil), FOLEX
(methotrexate), FOLEX PFS (methotrexate), FOLFIRI,
FOLFIRI-BEVACIZUMAB, FOLFIRI-CETUXIMAB, FOLFIRINOX, FOLFOX, FU-LV,
GARDASIL (recombinant human papillomavirus (HPV) quadrivalent
vaccine), GEMCITABINE-CISPLATIN, GEMCITABINE-OXALIPLATIN, GEMZAR
(gemcitabine hydrochloride), GILOTRIF (afatinib dimaleate), GLEEVEC
(imatinib mesylate), GLIADEL (carmustine implant), GLIADEL WAFER
(carmustine implant), HERCEPTIN (trastuzumab), HYCAMTIN (topotecan
hydrochloride), IFEX (ifosfamide), IFOSFAMIDUM (ifosfamide), INLYTA
(axitinib), INTRON A (recombinant interferon alfa-2b), IRESSA
(gefitinib), IXEMPRA (ixabepilone), JAKAFI (ruxolitinib phosphate),
JEVTANA (cabazitaxel), KADCYLA (ado-trastuzumab emtansine),
KEYTRUDA (pembrolizumab), KYPROLIS (carfilzomib), LIPODOX
(doxorubicin hydrochloride liposome), LUPRON (leuprolide acetate),
LUPRON DEPOT (leuprolide acetate), LUPRON DEPOT-3 MONTH (leuprolide
acetate), LUPRON DEPOT-4 MONTH (leuprolide acetate), LUPRON
DEPOT-PED (leuprolide acetate), MEGACE (megestrol acetate),
MEKINIST (trametinib), METHAZOLASTONE (temozolomide), METHOTREXATE
LPF (methotrexate), MEXATE (methotrexate), MEXATE-AQ
(methotrexate), MITOXANTRONE HYDROCHLORIDE, MITOZYTREX (mitomycin
c), MOZOBIL (plerixafor), MUSTARGEN (mechlorethamine
hydrochloride), MUTAMYCIN (mitomycin c), MYLOSAR (azacitidine),
NAVELBINE (vinorelbine tartrate), NEOSAR (cyclophosphamide),
NEXAVAR (sorafenib tosylate), NOLVADEX (tamoxifen citrate),
NOVALDEX (tamoxifen citrate), OFF, PAD, PARAPLAT (carboplatin),
PARAPLATIN (carboplatin), PEG-INTRON (peginterferon alfa-2b),
PEMETREXED DISODIUM, PERJETA (pertuzumab), PLATINOL (cisplatin),
PLATINOL-AQ (cisplatin), POMALYST (pomalidomide), prednisone,
PROLEUKIN (aldesleukin), PROLIA (denosumab), PROVENGE
(sipuleucel-t), REVLIMID (lenalidomide), RUBIDOMYCIN (daunorubicin
hydrochloride), SPRYCEL (dasatinib), STIVARGA (regorafenib), SUTENT
(sunitinib malate), SYLATRON (peginterferon alfa-2b), SYLVANT
(siltuximab), SYNOVIR (thalidomide), TAC, TAFINLAR (dabrafenib),
TARABINE PFS (cytarabine), TARCEVA (erlotinib hydrochloride),
TASIGNA (nilotinib), TAXOL (paclitaxel), TAXOTERE (docetaxel),
TEMODAR (temozolomide), THALOMID (thalidomide), TOPOSAR
(etoposide), TORISEL (temsirolimus), TPF, TRISENOX (arsenic
trioxide), TYKERB (lapatinib ditosylate), VECTIBIX (panitumumab),
VEIP, VELBAN (vinblastine sulfate), VELCADE (bortezomib), VELSAR
(vinblastine sulfate), VEPESID (etoposide), VIADUR (leuprolide
acetate), VIDAZA (azacitidine), VINCASAR PFS (vincristine sulfate),
VOTRIENT (pazopanib hydrochloride), WELLCOVORIN (leucovorin
calcium), XALKORI (crizotinib), XELODA (capecitabine), XELOX, XGEVA
(denosumab), XOFIGO (radium 223 dichloride), XTANDI (enzalutamide),
YERVOY (ipilimumab), ZALTRAP (ziv-aflibercept), ZELBORAF
(vemurafenib), ZOLADEX (goserelin acetate), ZOMETA (zoledronic
acid), ZYKADIA (ceritinib), ZYTIGA (abiraterone acetate), or a
combination thereof. In certain embodiments, the additional
pharmaceutical agent is a kinase inhibitor. In certain embodiments,
the additional pharmaceutical agent is a protein kinase inhibitor
(e.g., tyrosine protein kinase inhibitor). In certain embodiments,
the additional pharmaceutical agent is an inhibitor of a Src family
kinase. In certain embodiments, the additional pharmaceutical agent
is a CDK inhibitor. In certain embodiments, the additional
pharmaceutical agent is a CDK7 inhibitor. In certain embodiments,
the additional pharmaceutical agent is a CDK12 inhibitor. In
certain embodiments, the additional pharmaceutical agent is a CDK13
inhibitor. In certain embodiments, the additional pharmaceutical
agent is an inhibitor of one or more protein kinases selected from
the group consisting of IRAK1, IRAK4, BMX, and PI3K. In certain
embodiments, the additional pharmaceutical agent is an inhibitor of
one or more protein kinases selected from the group consisting of
BUB1B, CDK2, CDK9, CHEK2, FGR, HIPK4, PRKCQ, RET, SRC, and MELK. In
certain embodiments, the additional pharmaceutical agent is an
inhibitor of one or more protein kinases selected from the group
consisting of ABL, ARG, BLK, CSK, EphB1, EphB2, FOR, FRK, FYN, SRC,
YES, LCK, LYN, MAP2K5, NLK, p38a, SNRK, and TEC. In certain
embodiments, the additional pharmaceutical agent is an inhibitor of
one or more protein kinases selected from the group consisting of
ABL1(H396P)-phosphorylated, ABL1-phosphorylated, BLK, EPHA4, EPHB2,
EPHB3, EPHB4, FOR, JAK3(JH1domain-catalytic), KIT, KIT(L576P),
KIT(V559D), PDGFRB, SRC, YES, ABL1(H396P)-nonphosphorylated,
ABL1(Y253F)-phosphorylated, ABL1-nonphosphorylated, FRK, LYN,
ABL1(Q252H)-nonphosphorylated, DDR1, EPHB1, ERBB4, p38-alpha, ABL2,
ABL1(Q252H)-phosphorylated, SIK, EPHA8, MEK5,
ABL1(E255K)-phosphorylated, ABL1(F317L)-nonphosphorylated, FYN,
LCK, EPHA2, ABL1(M351T)-phosphorylated, TXK, EGFR(L858R),
EGFR(L861Q), ERBB2, ERBB3, EPHA5, ABL1(F317I)-nonphosphorylated,
EGFR(L747-E749del, A750P), CSK, EPHA1, ABL1(F317L)-phosphorylated,
BRAF(V600E), EGFR, KIT-autoinhibited, and EGFR(E746-A750del). In
certain embodiments, the additional pharmaceutical agent is an
inhibitor of one or more protein kinases selected from the group
consisting of ABL1 (F317L)-nonphosphorylated, ABL1
(H396P)-nonphosphorylated, ABL1 (H396P)-phosphorylated,
ABL1-phosphorylated, BLK, EPHA4, EPHB2, EPHB3, EPHB4,
JAK3(JH1domain-catalytic), KIT, KIT(L576P), KIT(V559D), LYN,
PDGFRB, SRC, YES, ABL1-nonphosphorylated,
ABL1(Y253F)-phosphorylated, ERBB3, FOR, FRK, p38-alpha,
ABL1(F317I)-nonphosphorylated, DDR1, EPHA2,
ABL1(Q252H)-phosphorylated, MEK5, ABL1(Q252H)-nonphosphorylated,
ABL2, FYN, EPHB1, ABL1 (E255K)-phosphorylated,
ABL1(F317L)-phosphorylated, EPHA1, ABL1(M351T)-phosphorylated,
ERBB4, TXK, LCK, EPHA8, SIK, EPHA5, EGFR(L861Q),
CSF1R-autoinhibited, BRAF(V600E), BRK, CSK, KIT(D816V),
KIT-autoinhibited, EGFR(L747-T751del,Sins), EGFR(L858R),
EGFR(L747-E749del, A750P), and CSF1R. In certain embodiments, the
additional pharmaceutical agent is an anti-angiogenesis agent,
anti-inflammatory agent, immunosuppressant, anti-bacterial agent,
anti-viral agent, cardiovascular agent, cholesterol-lowering agent,
anti-diabetic agent, anti-allergic agent, pain-relieving agent, or
a combination thereof. In certain embodiments, the compounds
described herein or pharmaceutical compositions can be administered
or used in combination with an anti-cancer therapy including, but
not limited to, transplantation (e.g., bone marrow transplantation,
stem cell transplantation), surgery, radiation therapy,
immunotherapy, and chemotherapy.
[0391] Also encompassed by the disclosure are kits (e.g.,
pharmaceutical packs). The kits provided may comprise a
pharmaceutical composition or compound described herein and a
container (e.g., a vial, ampule, bottle, syringe, and/or dispenser
package, or other suitable container). In some embodiments,
provided kits may optionally further include a second container
comprising a pharmaceutical excipient for dilution or suspension of
a pharmaceutical composition or compound described herein. In some
embodiments, the pharmaceutical composition or compound described
herein provided in the first container and the second container are
combined to form one unit dosage form.
Methods of Treatment and Uses
[0392] The present invention also provides methods for the
treatment or prevention of a proliferative disease (e.g., cancers
(e.g., leukemia, acute lymphoblastic leukemia, lymphoma, Burkitt's
lymphoma, melanoma, multiple myeloma, breast cancer, Ewing's
sarcoma, osteosarcoma, brain cancer, neuroblastoma, lung cancer,
colorectal cancer), benign neoplasms, diseases associated with
angiogenesis, inflammatory diseases, autoinflammatory diseases, and
autoimmune diseases).
[0393] The compounds described herein may exhibit kinase inhibitory
activity; the ability to inhibit cyclin-dependent kinase (CDK); the
ability to inhibit cyclin-dependent kinase 7 (CDK7); the ability to
inhibit cyclin-dependent kinase 7 (CDK7), without inhibiting
another cyclin-dependent kinase (CDK); the ability to inhibit
cyclin-dependent kinase 12 (CDK12); the ability to inhibit
cyclin-dependent kinase 12 (CDK12), without inhibiting another
cyclin-dependent kinase (CDK); the ability to inhibit
cyclin-dependent kinase 13 (CDK13); the ability to inhibit
cyclin-dependent kinase 13 (CDK13), without inhibiting another
cyclin-dependent kinase (CDK); the ability to inhibit
cyclin-dependent kinases 12 and 13 (CDK12 and CDK13); the ability
to inhibit cyclin-dependent kinases 12 and 13 (CDK12 and CDK13),
without inhibiting another cyclin-dependent kinase (CDK); a
therapeutic effect and/or preventative effect in the treatment of
cancers; a therapeutic effect and/or preventative effect in the
treatment of Myc-dependent cancers; and/or a therapeutic profile
(e.g., optimum safety and curative effect) that is superior to
existing chemotherapeutic agents.
[0394] Without wishing to be bound by any particular theory, the
compounds described herein are able to bind (e.g., covalently
modify) the protein kinase being inhibited. In certain embodiments,
the R.sup.2 group of a compound described herein is able to bind
(e.g., covalently modify) to the protein kinase. In certain
embodiments, the R.sup.2 group of a compound described herein is
able to covalently bind a cysteine residue of the protein kinase.
In certain embodiments, the compound is capable of covalently
modifying CDK7 (e.g., Cys312 of CDK7). In certain embodiments, the
R.sup.2 group of a compound described herein is able to covalently
modify residue Cys312 of CDK7. In certain embodiments, the compound
is capable of covalently modifying CDK12 (e.g., Cys1039 of CDK12).
In certain embodiments, the R.sup.2 group of a compound described
herein is able to covalently modify residue Cys1039 of CDK12. In
certain embodiments, the compound is capable of covalently
modifying CDK13 (e.g., Cys1017 of CDK13). In certain embodiments,
the R.sup.2 group of a compound described herein is able to
covalently modify residue Cys1017 of CDK13.
[0395] In another aspect, the present disclosure provides methods
of inhibiting the activity of a protein kinase in a subject, the
methods comprising administering to the subject an effective amount
(e.g., therapeutically effective amount) of a compound, or
pharmaceutical composition thereof, as described herein.
[0396] In another aspect, the present disclosure provides methods
of inhibiting the activity of a protein kinase in a biological
sample, the methods comprising contacting the biological sample
with an effective amount of a compound, or pharmaceutical
composition thereof, as described herein.
[0397] In another aspect, the present disclosure provides methods
of inhibiting the activity of a protein kinase in a tissue, the
methods comprising contacting the tissue with an effective amount
of a compound, or pharmaceutical composition thereof, as described
herein.
[0398] In another aspect, the present disclosure provides methods
of inhibiting the activity of a protein kinase in a cell, the
methods comprising contacting the cell with an effective amount of
a compound, or pharmaceutical composition thereof, as described
herein.
[0399] In certain embodiments, the subject being treated is a
mammal. In certain embodiments, the subject is a human. In certain
embodiments, the subject is a domesticated animal, such as a dog,
cat, cow, pig, horse, sheep, or goat. In certain embodiments, the
subject is a companion animal such as a dog or cat. In certain
embodiments, the subject is a livestock animal such as a cow, pig,
horse, sheep, or goat. In certain embodiments, the subject is a zoo
animal. In another embodiment, the subject is a research animal
such as a rodent, dog, or non-human primate. In certain
embodiments, the subject is a non-human transgenic animal such as a
transgenic mouse or transgenic pig.
[0400] In certain embodiments, the biological sample being
contacted with the compound or composition is breast tissue, bone
marrow, lymph node, lymph tissue, spleen, or blood.
[0401] In certain embodiments, the cell being contacted with the
compound or composition is in vitro. In certain embodiments, the
cell being contacted with the compound or composition is in vivo.
In certain embodiments, the cell being contacted with the compound
or composition is ex vivo. In certain embodiments, the cell being
contacted with the compound or composition is a malignant cell
(e.g., malignant blood cell). In certain embodiments, the cell
being contacted with the compound or composition is a malignant
hematopoietic stem cell (e.g., malignant myeloid cell or malignant
lymphoid cell). In certain embodiments, the cell being contacted
with the compound or composition is a malignant lymphocyte (e.g.,
malignant T-cell or malignant B-cell). In certain embodiments, the
cell being contacted with the compound or composition is a
malignant red blood cell, malignant white blood cell, or malignant
platelet. In certain embodiments, the cell being contacted with the
compound or composition is a malignant neutrophil, malignant
macrophage, or malignant plasma cell. In certain embodiments, the
cell being contacted with the compound or composition is a
carcinoma cell. In certain embodiments, the cell being contacted
with the compound or composition is a carcinoma breast cell. In
certain embodiments, the cell being contacted with the compound or
composition is a sarcoma cell. In certain embodiments, the cell
being contacted with the compound or composition is a sarcoma cell
from breast tissue.
[0402] The proliferative disease to be treated or prevented using
the compounds described herein may be associated with
overexpression of a kinase, such as cyclin-dependent kinase (CDK).
The process of eukaryotic cell division may be broadly divided into
a series of sequential phases termed G1, S, G2, and M. Correct
progression through the various phases of the cell cycle has been
shown to be critically dependent upon the spatial and temporal
regulation of a family of proteins known as cyclin dependent
kinases (CDKs) and a diverse set of their cognate protein partners
termed cyclins. CDKs are CDC2 (also known as CDK1) homologous
serine-threonine kinase proteins that are able to utilize ATP as a
substrate in the phosphorylation of diverse polypeptides in a
sequence-dependent context. Cyclins are a family of proteins
characterized by a homology region, containing approximately 100
amino acids, termed the "cyclin box" which is used in binding to,
and defining selectivity for, specific CDK partner proteins.
[0403] Modulation of the expression levels, degradation rates,
protein levels, and activity levels of various CDKs and cyclins
throughout the cell cycle leads to the cyclical formation of a
series of CDK/cyclin complexes, in which the CDKs are enzymatically
active. The formation of these complexes controls passage through
discrete cell cycle checkpoints and thereby enables the process of
cell division to continue. Failure to satisfy the prerequisite
biochemical criteria at a given cell cycle checkpoint, i.e.,
failure to form a required CDK/cyclin complex, can lead to cell
cycle arrest and/or cellular apoptosis. Aberrant cellular
proliferation can often be attributed to loss of correct cell cycle
control. Inhibition of CDK enzymatic activity therefore provides a
means by which abnormally dividing cells can have their division
arrested and/or be killed. The diversity of CDKs, and CDK
complexes, and their critical roles in mediating the cell cycle,
provides a broad spectrum of potential therapeutic targets selected
on the basis of a defined biochemical rationale.
[0404] CDK7, a member of the CDK family, was originally isolated as
the catalytic subunit of the trimeric CDK-activating kinase (CAK)
complex. This complex, consisting of CDK7, cyclin H, and MAT1, is
responsible for activation of the mitotic promoting factor in
vitro. The discovery that CDK7 was also a component of the basal
transcription repair factor IIH (TFIIH) implicated a dual role for
CDK7 in transcription as part of TFIIH and in the control of the
cell cycle as the trimeric CAK complex. TFIIH is a multi-subunit
protein complex identified as a factor required for RNA polymerase
II (RNAP 1)-catalyzed transcription, and subsequently this complex
was found to play a key role in nucleotide excision repair. CDK7 is
a component of at least three complexes, i.e., the trimeric CAK
complex, the quaternary complex with the XPD (or ERCC2, a protein
involved in transcription-coupled nucleotide excision repair), and
the nine-subunit TFIIH complex. The two functions of CDK7 in CAK
and CTD phosphorylation support critical facets of cellular
proliferation, cell cycling, and transcription. Overexpression of
CDK7 may inhibit apoptosis, promote transcription and cell
proliferation, and/or disrupt DNA repair, and therefore, cause
proliferative diseases. In certain embodiments, the proliferative
disease to be treated or prevented using the compounds described
herein may be associated with overexpression of a CDK (e.g.,
CDK7).
[0405] Cdk12 and Cdk13 are Cdc2-related proteins that share 92%
identity in their kinase domains (Chen et al., Exp. Neurol., 2014,
261, 10-21). CDK12 plays a critical role in cell processes, for
example, regulating transcription and splicing machinery by
stabilizing the RNAPII and DNA interaction, and regulating DNA
damage response (DDR) and maintenance of genomic stability by
modulating the expression of DDR genes. Overexpression of CDK12 has
been found to correlate, both at the transcriptional and protein
level, with pathological parameters of breast cancer disease.
[0406] A proliferative disease may be associated with aberrant
activity of a CDK (e.g., CDK7, CDK12, and/or CDK13). Aberrant
activity of a CDK (e.g., CDK7, CDK12, and/or CDK13) may be an
elevated and/or an inappropriate activity of the CDK. Deregulation
of cell cycle progression is a characteristic of a proliferative
disease, and a majority of proliferative diseases have
abnormalities in some component of CDK (e.g., CDK7, CDK12, and/or
CDK13) activity, frequently through elevated and/or inappropriate
CDK activation. Inhibition of the catalytic activity of CDK7,
CDK12, and/or CDK13 would be expected to inhibit cell cycle
progression by blocking the phosphorylation of cell cycle CDKs, and
would additionally inhibit transcription of effectors of cell
division. In certain embodiments, CDK7 is not overexpressed, and
the activity of CDK7 is elevated and/or inappropriate. In certain
other embodiments, CDK7 is overexpressed, and the activity of CDK7
is elevated and/or inappropriate. In certain embodiments, CDK12 is
not overexpressed, and the activity of CDK12 is elevated and/or
inappropriate. In certain embodiments, CDK12 is overexpressed, and
the activity of CDK12 is elevated and/or inappropriate. In certain
other embodiments, CDK13 is not overexpressed, and the activity of
CDK13 is elevated and/or inappropriate. In certain other
embodiments, CDK13 is overexpressed, and the activity of CDK13 is
elevated and/or inappropriate. The compounds described herein, and
pharmaceutically acceptable salts, solvates, hydrates, polymorphs,
co-crystals, tautomers, stereoisomers, isotopically labeled
derivatives, prodrugs, and compositions thereof, may inhibit the
activity of CDK7 and be useful in treating and/or preventing
proliferative diseases. The compounds described herein, and
pharmaceutically acceptable salts, solvates, hydrates, polymorphs,
co-crystals, tautomers, stereoisomers, isotopically labeled
derivatives, prodrugs, and compositions thereof, may inhibit the
activity of CDK12 and/or CDK13 and be useful in treating and/or
preventing proliferative diseases.
[0407] A proliferative disease may also be associated with
inhibition of apoptosis of a cell in a biological sample or
subject. All types of biological samples described herein or known
in the art are contemplated as being within the scope of the
invention. Apoptosis is the process of programmed cell death.
Inhibition of apoptosis may result in uncontrolled cell
proliferation and, therefore, may cause proliferative diseases. The
cell cycle CDKs (CDK1, 2, 4, and 6) are activated by
phosphorylation by CDK7/cyclin H (also called CAK). Inhibition of
CDK7 would therefore result in cell-cycle arrest at multiple points
in the cell cycle due to failure to activate the cell cycle CDKs.
CDK7 activates transcription by phosphorylating the CTD of RNAP II.
Inhibition of CTD phosphorylation has been shown to inhibit
transcription and reduce expression of short lived proteins,
including those involved in apoptosis regulation. It is appreciated
in the art that stalling of RNA polymerase may activate p53 (also
known as protein 53 or tumor protein 53, a tumor suppressor protein
that is encoded in humans by the TP53 gene), leading to apoptosis.
Thus, inhibition of the activity of CDK7 are expected to cause
cytotoxicity by inducing apoptosis. The compounds described herein,
and pharmaceutically acceptable salts, solvates, hydrates,
polymorphs, co-crystals, tautomers, stereoisomers, isotopically
labeled derivatives, prodrugs, and compositions thereof, may induce
apoptosis, and therefore, be useful in treating and/or preventing
proliferative diseases.
[0408] The CycK/Cdk12 complex regulates phosphorylation of Ser2 in
the C-terminal domain of RNA polymerase II and expression of a
small subset of human genes, as revealed in expression microarrays.
Through regulation of expression of DNA damage response genes (i.e.
oncogenes), CycK/Cdk12 protects cells from genomic instability. In
certain embodiments, the DNA damage response genes are BRCA1,
BRCA2, HER1, HER2, ATR, FANCI, or FANCD2. In certain embodiments,
the DNA damage response genes are BRCA1, HER2, ATR, FANCI, and
FANCD2. In certain embodiments, the DNA damage response genes are
BRCA1. In certain embodiments, the DNA damage response genes are
HER2.
[0409] In certain embodiments, the proliferative disease to be
treated or prevented using the compounds described herein is
cancer. All types of cancers disclosed herein or known in the art
are contemplated as being within the scope of the invention. In
certain embodiments, the proliferative disease is a cancer
associated with BCL-2 anti-apoptotic proteins (e.g., MCL-1 and/or
XIAP) (e.g., cancer associated with dependence on BCL-2
anti-apoptotic proteins). In certain embodiments, the proliferative
disease is a cancer associated with overexpression of MYC (a gene
that codes for a transcription factor). In certain embodiments, the
cancer is a MYC-dependent cancer. In certain embodiments, the
proliferative disease is a cancer associated with amplification of
BRCA1. In certain embodiments, the proliferative disease is a
cancer associated with amplification of HER2. In certain
embodiments, the proliferative disease is a hematological
malignancy. In certain embodiments, the proliferative disease is a
blood cancer. In certain embodiments, the proliferative disease is
a hematological malignancy. In certain embodiments, the
proliferative disease is leukemia. In certain embodiments, the
proliferative disease is chronic lymphocytic leukemia (CLL). In
certain embodiments, the proliferative disease is acute
lymphoblastic leukemia (ALL). In certain embodiments, the
proliferative disease is T-cell acute lymphoblastic leukemia
(T-ALL). In certain embodiments, the proliferative disease is
chronic myelogenous leukemia (CML). In certain embodiments, the
proliferative disease is acute myelogenous leukemia (AML). In
certain embodiments, the proliferative disease is acute monocytic
leukemia (AMoL). In certain embodiments, the proliferative disease
is lymphoma. In some embodiments, the proliferative disease is
Burkitt's lymphoma. In certain embodiments, the proliferative
disease is a Hodgkin's lymphoma. In certain embodiments, the
proliferative disease is a non-Hodgkin's lymphoma. In certain
embodiments, the proliferative disease is multiple myeloma. In
certain embodiments, the proliferative disease is melanoma. In
certain embodiments, the proliferative disease is colorectal
cancer. In certain embodiments, the proliferative disease is breast
cancer. In certain embodiments, the proliferative disease is
recurring breast cancer. In certain embodiments, the proliferative
disease is mutant breast cancer. In certain embodiments, the
proliferative disease is HER2+ breast cancer. In certain
embodiments, the proliferative disease is HER2-breast cancer. In
certain embodiments, the proliferative disease is triple-negative
breast cancer (TNBC). In certain embodiments, the proliferative
disease is a bone cancer. In certain embodiments, the proliferative
disease is osteosarcoma. In certain embodiments, the proliferative
disease is Ewing's sarcoma. In some embodiments, the proliferative
disease is a brain cancer. In some embodiments, the proliferative
disease is neuroblastoma. In some embodiments, the proliferative
disease is a lung cancer. In some embodiments, the proliferative
disease is small cell lung cancer (SCLC). In some embodiments, the
proliferative disease is non-small cell lung cancer. In some
embodiments, the proliferative disease is a benign neoplasm. All
types of benign neoplasms disclosed herein or known in the art are
contemplated as being within the scope of the invention. In some
embodiments, the proliferative disease is associated with
angiogenesis. All types of angiogenesis disclosed herein or known
in the art are contemplated as being within the scope of the
invention. In certain embodiments, the proliferative disease is an
inflammatory disease. All types of inflammatory diseases disclosed
herein or known in the art are contemplated as being within the
scope of the invention. In certain embodiments, the inflammatory
disease is rheumatoid arthritis.
In certain embodiments, the proliferative disease is an acute
inflammatory disease. In certain embodiments, the acute
inflammatory disease is rheumatoid arthritis, chron's disease, or
fibrosis. In some embodiments, the proliferative disease is an
autoinflammatory disease. All types of autoinflammatory diseases
disclosed herein or known in the art are contemplated as being
within the scope of the invention. In some embodiments, the
proliferative disease is an autoimmune disease. All types of
autoimmune diseases disclosed herein or known in the art are
contemplated as being within the scope of the invention.
[0410] Another aspect of the invention relates to methods of
inhibiting the activity of a kinase in a biological sample, tissue,
cell, or subject. In certain embodiments, the kinase is a CDK. In
certain embodiments, the kinase is CDK7. In certain embodiments,
the kinase is CDK12. In certain embodiments, the kinase is CDK13.
In certain embodiments, the activity of the kinase is aberrant
activity of the kinase. In certain embodiments, the activity of the
kinase is increased activity of the kinase. In certain embodiments,
the inhibition of the activity of the kinase is irreversible. In
other embodiments, the inhibition of the activity of the kinase is
reversible. In certain embodiments, the methods of inhibiting the
activity of the kinase include attaching a compound described
herein to the kinase.
[0411] Also provided in the present invention are methods of
inhibiting transcription of genes in a biological sample or
subject. In certain embodiments, the transcription of genes
affected by the activity of CDK7 may be inhibited by a compound of
the invention. In certain embodiments, the genes which may have
their transcription inhibited by the activity of CDK7 are one or
more selected from the group consisting of MYC, RUNX1, MYB, TAL1,
GATA3, KLF2, HNRPDL, p21, ASCL1, MYCN, INSM1, NEUROD1, NEUROG1,
FOXG1, FOXA1, SOX2, SOX4, BCL11A, OTX2, GAT2, PHOX2B, PLK2, TAF1,
CTGF, WEE1, SDIM, JUN, PIM1, IL8, and FOS1. In certain embodiments,
the genes which may have their transcription inhibited by the
activity of CDK7 include MYC, KLF2, E2F2, CDK6, CCND3, E2F3,
HNRPDL, TET1, IL7R, BRCA1, BRCA2, HER1, and HER2. In certain
embodiments, the transcription of genes affected by the activity of
CDK12 may be inhibited by a compound of the invention. In certain
embodiments, the genes which may have their transcription inhibited
by the activity of CDK12 are one or more selected from the group
consisting of BRCA1, FANCI, ATR, FANCD2, APEX1, NEK9, CHEK1, CHEK2,
ATM, RAD51C, RAD51D, ORC3L, MDC1, TERF2, ERCC4, FANCF, PARP9,
RUNX1, MYB, TAL1, MCL1, MYC, BCL2, ETS1, and EWS-FLI. In certain
embodiments, the transcription of genes affected by the activity of
CDK13 may be inhibited by a compound of the invention. In certain
embodiments, the gene is SNORA38.
[0412] The present invention also provides methods of inhibiting
cell growth in a biological sample, tissue, cell, or subject.
[0413] In still another aspect, the present invention provides
methods of inducing apoptosis of a cell in a biological sample,
tissue, cell, or subject.
[0414] In certain embodiments, the methods described herein include
administering to a subject or contacting a biological sample with
an effective amount of a compound described herein, or a
pharmaceutically acceptable salt, solvate, hydrate, polymorph,
co-crystal, tautomer, stereoisomer, isotopically labeled
derivative, or prodrug thereof, or a pharmaceutical composition
thereof. In certain embodiments, the methods described herein
include administering to a subject or contacting a biological
sample with an effective amount of a compound described herein, or
a pharmaceutically acceptable salt thereof, or a pharmaceutical
composition thereof. In certain embodiments, the compound is
contacted with a biological sample. In certain embodiments, the
compound is administered to a subject. In certain embodiments, the
compound is administered in combination with one or more additional
pharmaceutical agents described herein. The additional
pharmaceutical agent may be an anti-proliferative agent. In certain
embodiments, the additional pharmaceutical agent is an anti-cancer
agent. The additional pharmaceutical agent may also be a kinase
inhibitor. In certain embodiments, the additional pharmaceutical
agent is an inhibitor of a CDK. In certain embodiments, the
additional pharmaceutical agent is an inhibitor of CDK7. In certain
embodiments, the additional pharmaceutical agent is a selective
inhibitor of CDK7. In certain embodiments, the additional
pharmaceutical agent is a nonselective inhibitor of CDK7. In
certain embodiments, the additional pharmaceutical agent is an
inhibitor of CDK12. In certain embodiments, the additional
pharmaceutical agent is a selective inhibitor of CDK12. In certain
embodiments, the additional pharmaceutical agent is a nonselective
inhibitor of CDK12. In certain embodiments, the additional
pharmaceutical agent is an inhibitor of CDK13. In certain
embodiments, the additional pharmaceutical agent is a selective
inhibitor of CDK13. In certain embodiments, the additional
pharmaceutical agent is a nonselective inhibitor of CDK13. In
certain embodiments, the additional pharmaceutical agent is an
inhibitor of another CDK. In certain embodiments, the additional
pharmaceutical agent is a selective inhibitor of another CDK. In
certain embodiments, the additional pharmaceutical agent is a
nonselective inhibitor of another CDK. In certain embodiments, the
additional pharmaceutical agent is flavopiridol, triptolide,
SNS-032 (BMS-387032), PHA-767491, PHA-793887, BS-181, (S)--CR8,
(R)--CR8, or NU6140. In certain embodiments, the additional
pharmaceutical agent is an inhibitor of a mitogen-activated protein
kinase (MAPK). In certain embodiments, the additional
pharmaceutical agent is an inhibitor of a glycogen synthase kinase
3 (GSK3). In certain embodiments, the additional pharmaceutical
agent is an inhibitor of an AGC kinase. In certain embodiments, the
additional pharmaceutical agent is an inhibitor of a
calmodulin-dependent kinase (CaM Kinase). In certain embodiments,
the additional pharmaceutical agent is an inhibitor of a casein
kinase 1. In certain embodiments, the additional pharmaceutical
agent is an inhibitor of a STE kinase. In certain embodiments, the
additional pharmaceutical agent is an inhibitor of a tyrosine
kinase.
[0415] In some embodiments, the additional pharmaceutical agent is
a topoisomerase inhibitor, a MCL1 inhibitor, a BCL-2 inhibitor, a
BCL-xL inhibitor, a BRD4 inhibitor, a BRCA1 inhibitor, BRCA2
inhibitor, HER1 inhibitor, HER2 inhibitor, a CDK9 inhibitor, a
Jumonji histone demethylase inhibitor, or a DNA damage inducer. In
some embodiments, the additional pharmaceutical agent is etoposide,
obatoclax, navitoclax, JQ1,
4-(((5'-chloro-2'-(((1R,4R)-4-(((R)-1-methoxypropan-2-yl)amino)cyclohexyl-
)amino)-[2,4'-bipyridin]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitr-
ile, JIB04, or cisplatin. In some embodiments, the additional
pharmaceutical agent is etoposide, obatoclax, or navitoclax, and
the disease to be treated is breast cancer, e.g., triple-negative
breast cancer, HER2 positive breast cancer, HER2 negative breast
cancer, ER-positive breast cancer, ER-negative breast cancer, or
ER/PR-positive breast cancer. In some embodiments, the additional
pharmaceutical agent is etoposide, JIB04, or cisplatin, and the
disease to be treated is Ewing's sarcoma. In some embodiments, the
additional pharmaceutical agent is JQ1 or NVP2, and the disease to
be treated is leukemia, e.g., acute myelogenous leukemia,
myeloblastic leukemia, promyelocytic leukemia, myelomonocytic
leukemia, monocytic leukemia, monoblastic leukemia, or
megakaryoblastic leukemia. In certain embodiments, a pharmaceutical
composition described herein further comprises a combination of the
additional pharmaceutical agents described herein.
[0416] The inventive compounds or compositions may synergistically
augment inhibition of CDK7 induced by the additional pharmaceutical
agent(s) in the biological sample or subject. The inventive
compounds or compositions may synergistically augment inhibition of
CDK12 induced by the additional pharmaceutical agent(s) in the
biological sample or subject. The inventive compounds or
compositions may synergistically augment inhibition of CDK12 and/or
CDK13 induced by the additional pharmaceutical agent(s) in the
biological sample or subject. Thus, the combination of the
inventive compounds or compositions and the additional
pharmaceutical agent(s) may be useful in treating proliferative
diseases resistant to a treatment using the additional
pharmaceutical agent(s) without the inventive compounds or
compositions.
[0417] In some embodiments, the activity of a protein kinase is
non-selectively inhibited by the compounds or pharmaceutical
compositions described herein. In some embodiments, the activity of
the protein kinase being inhibited is selectively inhibited by the
compounds or pharmaceutical compositions described herein, compared
to the activity of a different protein (e.g., a different protein
kinase). In certain embodiments, the activity of CDK (e.g., CDK7,
CDK12, or CDK13) is selectively inhibited by a compound or
pharmaceutical composition described herein, compared to the
activity of a different protein. In certain embodiments, the
activity of CDK7 is selectively inhibited by a compound or
pharmaceutical composition described herein, compared to the
activity of a different CDK protein. In certain embodiments, the
activity of CDK7 is selectively inhibited by a compound or
pharmaceutical composition described herein, compared to the
activity of CDK12. In certain embodiments, the activity of CDK7 is
selectively inhibited by a compound or pharmaceutical composition
described herein, compared to the activity of CDK13. In certain
embodiments, the activity of CDK7 is selectively inhibited by a
compound or pharmaceutical composition described herein, compared
to the activity of CDK12 and the activity of CDK13. In certain
embodiments, the activity of CDK12 is selectively inhibited by a
compound or pharmaceutical composition described herein, compared
to the activity of CDK7. In certain embodiments, the activity of
CDK13 is selectively inhibited by a compound or pharmaceutical
composition described herein, compared to the activity of CDK7. In
certain embodiments, the activity of CDK12 and the activity of
CDK13 are selectively inhibited by a compound or pharmaceutical
composition described herein, compared to the activity of CDK7.
[0418] The selectivity of a compound or pharmaceutical composition
described herein in inhibiting the activity of a protein kinase
over a different protein (e.g., a different protein kinase) may be
measured by the quotient of the IC.sub.50value of the compound or
pharmaceutical composition in inhibiting the activity of the
different protein over the IC.sub.50 value of the compound or
pharmaceutical composition in inhibiting the activity of the
protein kinase. The selectivity of a compound or pharmaceutical
composition described herein for a protein kinase over a different
protein may also be measured by the quotient of the K.sub.d value
of an adduct of the compound or pharmaceutical composition and the
different protein over the K.sub.d value of an adduct of the
compound or pharmaceutical composition and the protein kinase. In
certain embodiments, the selectivity is at least 2-fold, at least
3-fold, at least 5-fold, at least 10-fold, at least 30-fold, at
least 100-fold, at least 300-fold, at least 1,000-fold, at least
3,000-fold, at least 10,000-fold, at least 30,000-fold, or at least
100,000-fold. In certain embodiments, the selectivity is not more
than 100,000-fold, not more than 10,000-fold, not more than
1,000-fold, not more than 100-fold, not more than 10-fold, or not
more than 2-fold. Combinations of the above-referenced ranges
(e.g., at least 2-fold and not more than 10,000-fold) are also
within the scope of the disclosure.
[0419] In certain embodiments, a kit described herein includes a
first container comprising a compound or pharmaceutical composition
described herein. In certain embodiments, a kit described herein is
useful in treating a proliferative disease (e.g., cancers (e.g.,
leukemia, acute lymphoblastic leukemia, lymphoma, Burkitt's
lymphoma, melanoma, multiple myeloma, breast cancer, Ewing's
sarcoma, osteosarcoma, brain cancer, neuroblastoma, lung cancer,
colorectal cancer), benign neoplasms, diseases associated with
angiogenesis, inflammatory diseases, autoinflammatory diseases, and
autoimmune diseases) in a subject in need thereof, preventing a
proliferative disease in a subject in need thereof, inhibiting the
activity of a protein kinase (e.g., CDK (e.g., CDK7, CDK12, or
CDK13)) in a subject, biological sample, tissue, or cell, and/or
inducing apoptosis in a cell.
[0420] In certain embodiments, a kit described herein further
includes instructions for using the compound or pharmaceutical
composition included in the kit. A kit described herein may also
include information as required by a regulatory agency such as the
U.S. Food and Drug Administration (FDA). In certain embodiments,
the information included in the kits is prescribing information. In
certain embodiments, the kits and instructions provide for treating
a proliferative disease in a subject in need thereof, preventing a
proliferative disease in a subject in need thereof, inhibiting the
activity of a protein kinase (e.g., CDK (e.g., CDK7, CDK12, or
CDK13)) in a subject, biological sample, tissue, or cell, and/or
inducing apoptosis in a cell. A kit described herein may include
one or more additional pharmaceutical agents described herein as a
separate composition.
EXAMPLES
[0421] In order that the invention described herein may be more
fully understood, the following examples are set forth. The
synthetic and biological examples described in this application are
offered to illustrate the compounds, pharmaceutical compositions,
and methods provided herein and are not to be construed in any way
as limiting their scope.
Pulldown Assay (FIGS. 2-4 and 7-9)
[0422] Jurkat cells were treated with DMSO or concentration of
compound indicated. 6 hours after treatment, cells were washed and
harvested by resuspending in lysis buffer (50 mM Hepes pH 7.4, 150
mM NaCl, 1% NP-40, 5 mM EDTA, protease and phosphatase inhibitors)
and lysing on ice 30 minutes. Lysates were cleared by
centrifugation at 15,000 rpm 30 minutes. Biotin-labeled THZ1 was
added to 1 .mu.M to lysates and rotated at 4.degree. C. overnight.
Streptavidin-agarose beads were washed and 30 .mu.L slurry was
added to each lysate and rotated for 1 hour at 4.degree. C. Beads
were washed 5 times with lysis buffer and 50 .mu.L 2.times.LDS
buffer was added to each sample. Samples were boiled and equal
volume of protein was loaded onto gel. Gel was transferred to
nitrocellulose and blotted for Cyclin K and Cyclin H.
Interpretation of Results
[0423] We conclude that pre-treatment of cells with several of the
compounds, but not DMSO, blocks biotin-THZ1 from being able to bind
to CDK12, which blocks the pulldown of CDK12 and CDK13-associated
Cyclin K. This indicates that active compounds are able to engage
CDK12 and CDK13-associated cyclin K complexes in cells and block
binding of these complexes by bio-THZ1. We do not see a similar
loss of pulldown of Cyclin H, indicating that these compounds are
not able to bind to CDK7-associated cyclin H complexes and block
its association with biotin-THZ1.
Growth Assay (FIG. 5)
[0424] Jurkat cells were plated at 30,000 cells/well and treated
with a titration of compounds indicated. Cells were allowed to grow
for 72 hours. Cells were assayed using CELLTITER GLO (Promega) to
determine cell viability by measuring the amount of ATP present,
which is an indicator of cell metabolic activity. Results are
graphed as luminescent values. Curves were generated using PRISM
and an IC.sub.50 value was determined.
Interpretation of Results
[0425] We can conclude that several of the compounds generated and
tested are more potent against cell growth than the parent SNS-032
compound in Jurkat cells.
Synthesis of the Compounds
[0426] The compounds provided herein can be prepared from readily
available starting materials using the following general methods
and procedures. Reactions were monitored by thin layer
chromatography (TLC) with 0.25 mm E. Merck pre-coated silica gel
plates (60 F.sub.2 4) and Waters LCMS system (Waters 2489
UV/Visible Detector, Waters 3100 Mass, Waters 515 HPLC pump, Waters
2545 Binary Gradient Module, Waters Reagent Manager, Waters 2767
Sample Manager) using SunFire.TM. C18 column (4.6.times.50 mm, 5 m
particle size): solvent gradient=95% A at 0 min, 0% A at 5 min;
solvent A=0.5% TFA in Water; solvent B=Methanol; flow rate: 1.5
mL/min. Purification of reaction products was carried out by flash
chromatography using CombiFlash.RTM. Rf with Teledyne Isco
RediSep.RTM. Rf High Performance Gold or Silicycle SiliaSep.TM.
High Performance columns (4 g, 12 g, 24 g, 40 g, 80 g or 120 g) or
by Waters preparative HPLC system with a C18 column: solvent
gradient=100% A at 0 min, 0% A at 15 min; solvent A=0.5% TFA in
Water; solvent B=Methanol; flow rate: 20 mL/min. The purity of all
compounds was over 95% and was analyzed with Waters LCMS system.
.sup.1H NMR and .sup.13C NMR spectra were obtained using a Varian
Inova-600 or 400 MHz spectrometer. Chemical shifts are reported
relative to chloroform (.delta.=7.24) for .sup.1H NMR or dimethyl
sulfoxide (.delta.=2.50) for .sup.1H NMR and dimethyl sulfoxide
(.delta.=39.51) for .sup.13C NMR. Data are reported as (br=broad,
s=singlet, d=doublet, t=triplet, q=quartet, m=multiplet).
##STR00240## ##STR00241##
5-Thiocyanatothiazol-2-amine (MFH-2-76-1)
[0427] A mixture of 2-amino-5-bromothiazole hydrobromide (6.0 g,
23.08 mmol) and potassium thiocyanate (9.0 g, 92.32 mmol) in
methanol (150 mL) was stirred at room temperature for 20 h.
Methanol was evaporated and water (20 ml) was added. The pH of the
aqueous solution was adjusted to pH=12 with 10% NaOH and then a
precipitate was formed. The solid was collected by filtration to
yield compound MFH-2-76-1 (1.5 g, 41%) as a solid. LCMS (m/z): 158
[M+H]+.
5-((5-tert-Butyloxazol-2-yl)methylthio)thiazol-2-amine
(MFH-2-80-1)
[0428] To a solution of compound MFH-2-76-1 (315 mg, 2 mmol) in
EtOH (5 ml) was added NaBH.sub.4 (151 mg, 4 mmol) at room
temperature. The mixture was stirred for 1 h, and then acetone (3
ml) was slowly added. After 1 h, a solution of
5-tert-Butyl-2-chloromethyloxazole (348 mg, 2 mmol) in EtOH (3 ml)
was added. The resulting dark reaction mixture was heated to reflux
for 1 h, and was then cooled and concentrated in reduced pressure.
The residue was then dissolved in ethyl acetate and washed with
water. The organic phase was separated, dried (MgSO.sub.4) and
concentrated in reduced pressure to give a crude product MFH-2-80-1
(310 mg, 57%) LCMS (m/z): 270 [M+H]+.
5-[[(5-t-buty2-oxazolyl)methyl]thio]-2-bromo (MFH-2-83-1)
[0429] To a solution of CuBr.sub.2 (1.8 g, 8 mmol) in acetonitrile
(48 mL) at 0.degree. C. was added t-BuONO (827 mg, 8 mmol) followed
by compound MFH-2-80-1 (1.8 g, 6.68 mmol). The mixture was stirred
at 0.degree. C. for 1 h and then was warmed up to room temperature.
Ethyl acetate was added and the organic mixture washed with
hydrochloric acid (50 mL), dried over magnesium sulfate, filtered
through a pad of silica gel, and concentrated in reduced pressure.
The residue was purified by chromatographed on silica gel to give
product. MFH-2-83-1 (1.5g, 67%). LCMS (m/z): 334 [M+H]+.
(R)-tert-butyl3-(5-((5-tert-butyloxazol-2-yl)methylthio)triazol-2-ylamino)-
piperidine-1-carboxylate (MFH-2-84-1)
[0430] The mixture of MFH-2-83-1 (1.5g, 4.5 mmol), (R)-tert-butyl
3-aminopiperidine-1-carboxylate (1.6 g, 8.1 mmol) and DIEA (1.5 g,
11.25 mmol) in NMP (5 mL) was stirred at 140.degree. C. for
overnight. The residue was extracted with chloroform and
iso-propanol (4:1). The organic phase was washed with brine (50 mL)
and dried over Na.sub.2SO.sub.4. After removal of the solvent, the
residue was purified by silica gel (MeOH/DCM=0-20%) to obtain
MFH-2-84-1 (1.36 g, yield 66.8%). LCMS (m/z): 453 [M+H].sup.+.
(R)-5-((5-tert-butyloxazol-2-yl)methylthio)-N-(piperidin-3-yl)thiazol-2-am-
ine (MFH-2-85-1)
[0431] To a solution of MFH-2-85-1 (1.36 g, 3 mmol) in methanol (18
mL) was added 4N HCl/dioxane (18 mL). The solution was then stirred
for 3h at room temperature and the solvent was removed under
reduced pressure to provide a crude which was directly used in the
next step. LCMS (m/z): 353 [M+H].sup.+.
[0432]
(R)-(3-(5-((5-tert-butyloxazol-2-yl)methylthi)triazol-2-ylamino)pip-
eridin-1-yl)(4-nitrophenyl)methanone (MFH-2-86-1)
[0433] The mixture of MFH-2-85-1 (250 mg, 0.643 mmol),
4-nitrobenzoyl chloride (132 mg, 0.71 mmol) in pyridine (3 mL) was
stirred for overnight at room temperature. Then the reaction
mixture was concentrated under reduced pressure and the residue was
directly used in the next step. LCMS (m/z): 502 [M+H].sup.+.
(R)-(4-aminophenyl)(3-(5-((5-tert-butyloxazol-2-yl)methylthio)thiazol-2-yl-
amino)piperidin-1-yl)methanone (MFH-2-89-1)
[0434] To a solution of MFH-2-86-1 (270 mg, 0.54 mmol) in ethyl
acetate and methanol (1:1) were added Tin (11) chloride dehydrate
(1.2 g, 5.4 mmol) and conc. HCl (0.2 mL). After stirring for 3 h at
80.degree. C., the reaction mixture was diluted with chloroform and
iso-propanol (4:1), neutralized with saturated NaHCO.sub.3 and
filtered. The filtrate was extracted with chloroform and
iso-propanol (4:1), concentrated under reduced pressure and the
resulting residue was purified by silica gel column chromatography
(MeOH/DCM=0-20%) to give MFH-2-89-1 (140 mg, yield 55%). LCMS
(m/z): 472 [M+H]+.
(R)--N-(4-(3-(5-((5-tert-butyloxazol-2-yl)methylthio)triazol-2-ylamino)pip-
eridine-1-carbonyl)phenyl)acrylamide (MFH-2-90-1)
[0435] To a solution of MFH-2-89-1 (140 mg, 0.30 mmol) and DIPEA
(0.2 mL) in CH.sub.3CN (2 mL) was added acryloyl chloride (35 mg,
0.39 mmol) in DCM (0.8 mL) dropwise. The mixture was then stirred
at 0.degree. C. for 1 h. The solution was then concentrated under
reduced pressure and the residue was purified by prep-HPLC
(MeOH/H.sub.2O, 0.05% TFA) to provide MFH-2-90-1 (24.7 mg, yield
16%). LCMS (m/z): 526 [M+H]+. 1H NMR (500 MHz, DMSO) .delta. 10.27
(s, 1H), 7.97 (s, 1H), 7.67 (s, 2H), 7.35 (d, J=6.7 Hz, 2H), 6.87
(d, J=26.3 Hz, 1H), 6.70 (s, 1H), 6.43 (dd, J=17.0, 10.1 Hz, 1H),
6.27 (dd, J=17.0, 1.8 Hz, 1H), 5.77 (dd, J=10.1, 1.8 Hz, 1H), 3.94
(s, 2H), 3.79 (s, 1H), 3.63 (d, J=23.9 Hz, 2H), 3.14 (d, J=65.1 Hz,
2H), 1.95 (s, 1H), 1.76 (s, 1H), 1.52 (d, J=7.8 Hz, 2H), 1.17 (s,
9H).
##STR00242## ##STR00243##
(S)-tert-butyl3-(5-((5-tert-butyloxazol-2-yl)methylthio)thiazol-2-ylamino-
)piperline-1-carboxylate (MFH-2-82-1)
[0436] The mixture of MFH-2-83-1 (150 mg, 0.45 mmol),
(S)-tert-butyl 3-aminopiperidine-1-carboxylate (160 mg, 0.81 mmol)
and DIEA (150 mg, 1.13 mmol) in NMP (1 mL) was stirred at
140.degree. C. for overnight. The residue was extracted with
chloroform and iso-propanol (4:1). The organic phase was washed
with brine (20 mL) and dried over Na.sub.2SO.sub.4. After removal
of the solvent, the residue was purified by silica gel
(MeOH/DCM=0-20%) to obtain MFH-2-82-1 (136 mg, yield 67%). LCMS
(m/z): 453 [M+H].sup.+.
(S)-5-((5-tert-butyloxazol-2-yl)methylthio)-N-(piperidin-3-yl)thiazol-2-am-
ine (MFH-2-87-1)
[0437] To a solution of MFH-2-82-1 (136 mg, 0.3 mmol) in methanol
(3 mL) was added 4N HCl/dioxane (3 mL). The solution was then
stirred for 3h at room temperature and the solvent was removed
under reduced pressure to provide a crude which was directly used
in the next step. LCMS (m/z): 353 [M+H].sup.+.
(S)-(3-(5-((5-tert-butyloxazol-2-yl)methylthio)thiazol-2-ylamino)piperidin-
-1-yl)(4-nitrophenyl)methanone (MFH-2-88-1)
[0438] The mixture of MFH-2-87-1 (250 mg, 0.643 mmol),
4-nitrobenzoyl chloride (132 mg, 0.71 mmol) in pyridine (3 mL) was
stirred for overnight at room temperature. Then the reaction
mixture was concentrated under reduced pressure and the residue was
directly used in the next step. LCMS (m/z): 502 [M+H].sup.+.
(S)-(4-aminophenyl)(3-(5-((5-tert-butyloxazol-2-yl)methylthio)thiazol-2-yl-
amino)piperidin-1-yl)methanone (MFH-2-91-1)
[0439] To a solution of MFH-2-88-1 (270 mg, 0.54 mmol) in ethyl
acetate and methanol (1:1) were added Tin(II) chloride dehydrate
(1.2 g, 5.4 mmol) and cone. HCl (0.2 mL). After stirring for 3 h at
80.degree. C., the reaction mixture was diluted with chloroform and
iso-propanol (4:1), neutralized with saturated NaHCO.sub.3 and
filtered. The filtrate was extracted with chloroform and
iso-propanol(4:1), concentrated under reduced pressure and the
resulting residue was purified by silica gel column chromatography
(MeOH/DCM=0-20%) to give MFH-2-91-1 (140 mg, yield 55%). LCMS
(m/z): 472 [M+H]+.
(S)--N-(4-(35-((5-tert-butyloxazol-2-yl)methylthio)thiazol-2-ylamino)piper-
idine-1-carbonyl)phenyl)acrylamide (MFH-2-92-1)
[0440] To a solution of MFH-2-91-1 (30 mg, 0.06 mmol) and DIPEA
(0.2 mL) in CH.sub.3CN (2 mL) was added acryloyl chloride (8 mg,
0.08 mmol) in DCM (0.5 mL) dropwise. The mixture was then stirred
at 0.degree. C. for 1h. The solution was then concentrated under
reduced pressure and the residue was purified by prep-HPLC
(MeOH/H.sub.2O, 0.05% TFA) to provide MFH-2-92-1 (11 mg, yield
33%). LCMS (m/z): 526 [M+H]+. 1H NMR (500 MHz, DMSO) .delta. 10.30
(s, 1H), 8.07 (s, 1H), 7.65 (s, 2H), 7.35 (d, J=6.7 Hz, 2H), 6.88
(d, J=37.7 Hz, 1H), 6.70 (s, 1H), 6.43 (dd, J=16.9, 10.1 Hz, 1H),
6.27 (dd, J=17.0, 1.6 Hz, 1H), 5.77 (dd, J=10.2, 1.7 Hz, 1H), 3.95
(s, 2H), 3.79 (s, 1H), 3.63 (d, J=23.9 Hz, 2H), 3.14 (d, J=65.1 Hz,
2H), 1.92 (s, J=18.7 Hz, 1H), 1.76 (s, 1H), 1.52 (d, J=6.6 Hz, 2H),
1.17 (s, 9H).
##STR00244##
(R)-tert-butyl4-3-(5-((5-tert-butyloxazol-2-yl)methylthio)thiazol-2-ylami-
no)piperidine-1-carbonyl)cyclohexylcarbamate (MFH-2-93-1)
[0441] The mixture of MFH-2-85-1 (116 mg, 0.33 mmol),
4-(tert-butoxycarbonylamino)cyclohexanecarboxylic acid (120 mg, 0.5
mmol), HOBT (68 mg, 0.5 mmol) and EDCI (96 mg, 0.5 mmol) in DCM (10
ml) was stirred for overnight. The reaction mixture was diluted
with DCM (25 ml). The organic phase was washed with saturated
Na.sub.2CO.sub.3 and brine (20 mL) and dried over Na.sub.2SO.sub.4.
After removal of the solvent, the residue was purified by silica
gel (MeOH/DCM=0-20%) to obtain MFH-2-93-1 (80 mg, yield 42%). LCMS
(m/z): 578 [M+H]+.
(R)-(4-aminocyclohexyl)(3-(5-((5-tert-butyloxazol-2-yl)methylthio)thiazol--
2-ylamino)piperidin-1-yl)methanone (MFH-2-94-1)
[0442] To a solution of MFH-2-93-1 (80 mg, 0.14 mmol) in methanol
(3 mL) was added 4N HCl/dioxane (3 mL). The solution was then
stirred for 3h at room temperature and the solvent was removed
under reduced pressure to provide a crude which was directly used
in the next step. LCMS (m/z): 478 [M+H].sup.+.
(R)--N-(4-(3-(5-(5-tert-butyloxazol-2-yl)methylthio)thiazol-2-ylamino)pipe-
ridine-1-carbonyl)cyclohexyl)acrylamide (MFH-2-95-1)
[0443] To a solution of MFH-2-94-1 (30 mg, 0.06 mmol) and DIPEA
(0.2 mL) in CH.sub.3CN (2 mL) was added acryloyl chloride (8 mg,
0.08 mmol) in DCM (0.5 mL) dropwise. The mixture was then stirred
at 0.degree. C. for 1h. The solution was then concentrated under
reduced pressure and the residue was purified by prep-HPLC
(MeOH/H.sub.2O, 0.05% TFA) to provide MFH-2-95-1 (15.9 mg, yield
48%). LCMS (m/z): 532 [M+H]+.
##STR00245##
(R)-tert-butyl4-(3-(5-((5-tert-butyloxazol-2-yl)methylthio)triazol-2-ylam-
ino)piperidine-1-carbonyl)benzylcarbamate (MFH-2-96-1)
[0444] The mixture of MFH-2-85-1 (88 mg, 0.25 mmol),
4-((tert-butoxycarbonylamino)methyl)benzoic acid (94 mg, 0.38
mmol), HOBT (51 mg, 0.38 mmol) and EDCI (72 mg, 0.38 mmol) in DCM
(8 ml) was stirred for overnight. The reaction mixture was diluted
with DCM (25 ml). The organic phase was washed with saturated
Na.sub.2CO.sub.3 and brine (20 mL) and dried over Na.sub.2SO.sub.4.
After removal of the solvent, the residue was purified by silica
gel (MeOH/DCM=0-20%) to obtain MFH-2-96-1 (125 mg, yield 85%). LCMS
(m/z): 586 [M+H]+.
(R)-(4-(aminomethyl)phenyl)(3-(5-((5-tert-butyloxazol-2-yl)methylthio)thia-
zol-2-ylamino)piperidin-1-yl)methanone (MFH-2-97-1)
[0445] To a solution of MFH-2-96-1 (125 mg, 0.21 mmol) in methanol
(3 mL) was added 4N HCl/dioxane (3 mL). The solution was then
stirred for 3h at room temperature and the solvent was removed
under reduced pressure to provide a crude which was directly used
in the next step. LCMS (m/z): 486 [M+H].sup.+.
(R)--N-(4-(3-(5-((5-tert-butyloxazol-2-yl)methylthio)thiazol-2-ylamino)pip-
eridine-1-carbonyl)benzyl)acrylamide (MFH-2-98-1)
[0446] To a solution of MFH-2-97-1 (50 mg, 0.1 mmol) and DIPEA (0.2
mL) in CH.sub.3CN (2 mL) was added acryloyl chloride (12 mg, 0.13
mmol) in DCM (0.5 mL) dropwise. The mixture was then stirred at
0.degree. C. for 1 h. The solution was then concentrated under
reduced pressure and the residue was purified by prep-HPLC
(MeOH/H.sub.2O, 0.05% TFA) to provide MFH-2-98-1 (16.4 mg, yield
30%). LCMS (m/z): 540 [M+H]+. .sup.1H NMR (500 MHz, DMSO) .delta.
8.62 (s, 1H), 8.08 (d, J=54.5 Hz, 1H), 7.28 (d, J=31.4 Hz, 4H),
6.91 (d, J=65.2 Hz, 1H), 6.72 (s, 1H), 6.28 (dd, J=17.1, 10.2 Hz,
1H), 6.13 (dd, J=17.1, 2.2 Hz, 1H), 5.62 (dd, J=10.2, 1.9 Hz, 1H),
4.36 (s, 2H), 3.95 (s, 2H), 3.67-3.62 (m, 2H), 3.33 (s, 1H),
3.14-2.83 (m, 2H), 1.94 (s, 1H), 1.74 (d, J=65.1 Hz, 1H), 1.53 (s,
2H), 1.20 (s, 9H).
##STR00246##
(R)-4-nitrophenyl3-(5-((5-tert-butyloxazol-2-yl)methylthio)thiazol-2-ylam-
ino)piperidine-1-carboxylate (MFH-2-99-1)
[0447] To a solution of MFH-2-85-1 (210 mg, 0.6 mmol) and DIPEA
(116 mg, 0.9 mmol) in DCM (8 mL) was added 4-nitrophenyl
chloroformate (133 mg, 0.66 mmol) in DCM (1 mL) dropwise. The
mixture was then stirred at 0.degree. C. for 1 h. The solution was
then concentrated under reduced pressure and the residue was
purified by silica gel (MeOH/DCM=0-20%) to obtain MFH-2-99-1 (283
mg, yield 91%). LCMS (m/z): 518 [M+H]+.
(R)-4-aminophenyl3-(5-((5-tert-butyloxazol-2-yl)methylthio)thiazol-2-ylami-
no)piperidine-1-carboxylate (MFH-2-100-1)
[0448] To a solution of MFH-2-99-1 (106 mg, 0.21 mmol) in ethyl
acetate and methanol (1:1) were added Tin(II) chloride dehydrate
(462 mg, 2.1 mmol) and conc. HCl (0.1 mL). After stirring for 3 h
at 80.degree. C., the reaction mixture was diluted with chloroform
and iso-propanol (4:1), neutralized with saturated NaHCO.sub.3 and
filtered. The filtrate was extracted with chloroform and
iso-propanol (4:1), concentrated under reduced pressure and the
resulting residue was purified by silica gel column chromatography
(MeOH/DCM=0-20%) to give MFH-2-100-1 (60 mg, yield 60%). LCMS
(m/z): 488 [M+H]+.
(R)-4-acrylamidophenyl3-(5-((5-tert-butyloxazol-2-yl)methylthio)thiazol-2--
ylamino)piperidine-1-carboxylate (MFH-2-102-1)
[0449] To a solution of MFH-2-100-1 (60 mg, 0.12 mmol) and DIPEA
(0.2 mL) in CH.sub.3CN (2 mL) was added acryloyl chloride (15 mg,
0.16 mmol) in DCM (0.5 mL) dropwise. The mixture was then stirred
at 0.degree. C. for 1h. The solution was then concentrated under
reduced pressure and the residue was purified by prep-HPLC
(MeOH/H.sub.2O, 0.05% TFA) to provide MFH-2-102-1 (18.1 mg, yield
27%). LCMS (m/z): 542 [M+H]+. .sup.1H NMR (500 MHz, DMSO) .delta.
10.17 (s, 1H), 8.13 (d, J=14.2 Hz, 1H), 7.64 (s, 2H), 7.08 (s, 1H),
7.01 (d, J=9.0 Hz, 1H), 6.95 (s, 1H), 6.72 (dd, J=11.3, 7.1 Hz,
1H), 6.42 (dd, J=16.9, 10.1 Hz, 1H), 6.25 (dd, J=17.0, 1.7 Hz, 1H),
5.82-5.68 (m, 1H), 3.94 (s, 2H), 3.84 (s, 1H), 3.74 (s, 1H), 3.26
(d, J=44.4 Hz, 2H), 2.93 (s, 1H), 1.92 (d, J=13.1 Hz, 1H), 1.79 (d,
J=6.3 Hz, 1H), 1.52 (s, 2H), 1.21 (s, 9H).
##STR00247##
(R)-tert-butyl1-(3-(5-(5-tert-butyloxazol-2-yl)methylthio)thiazol-2-ylami-
no)piperidine-1-carbonyl)piperidin-4-ylcarbamate (MFH-2-101-1)
[0450] The mixture of MFH-2-99-1 (177 mg, 0.34 mmol) and tert-butyl
piperidin-4-ylcarbamate (89 mg, 0.44 mmol) in DMSO (3 mL) was
stirred at 60.degree. C. overnight. The residue was extracted with
chloroform and iso-propanol (4:1). The organic phase was washed
with brine (20 mL) and dried over Na.sub.2SO.sub.4. After removal
of the solvent, the residue was purified by silica gel
(MeOH/DCM=0-20%) to obtain MFH-2-101-1 (150 mg, yield 76%). LCMS
(m/z): 579 [M+H].sup.+.
(R)-(4-aminopiperidin-1-yl)(3-(5-((5-tert-butyloxazol-2-yl)methylthio)thia-
zol-2-ylamino)piperidin-1-yl)methanone (MFH-2-103-1)
[0451] To a solution of MFH-2-101-1 (150 mg, 0.26 mmol) in methanol
(3 mL) was added 4N HCl/dioxane (3 mL). The solution was then
stirred for 3h at room temperature and the solvent was removed
under reduced pressure to provide a crude which was directly used
in the next step. LCMS (m/z): 479 [M+H].sup.+.
(R)--N-(1-(3-(5-((5-tert-butyloxazol-2-yl)methylthio)thiazol-2-ylamino)pip-
eridine-1-carbonyl)piperidin-4-yl)acrylamide (MFH-2-104-1)
[0452] To a solution of MFH-2-103-1 (30 mg, 0.06 mmol) and DIPEA
(0.2 mL) in CH.sub.3CN (2 mL) was added acryloyl chloride (8 mg,
0.08 mmol) in DCM (0.5 mL) dropwise. The mixture was then stirred
at 0.degree. C. for 1h. The solution was then concentrated under
reduced pressure and the residue was purified by prep-HPLC
(MeOH/H.sub.2O, 0.05% TFA) to provide MFH-2-104-1 (10.2 mg, yield
30%). LCMS (m/z): 533 [M+H]+. .sup.1H NMR (500 MHz, DMSO) .delta.
8.13 (s, 1H), 8.07 (d, J=7.7 Hz, 1H), 6.97 (s, 1H), 6.70 (d, J=5.5
Hz, 1H), 6.18 (dd, J=17.1, 10.1 Hz, 1H), 6.07 (dd, J=17.1, 2.2 Hz,
1H), 5.57 (dd, J=10.1, 2.2 Hz, 1H), 3.94 (s, 2H), 3.81-3.71 (m,
2H), 3.48 (s, 2H), 3.34-3.24 (m, 2H), 2.79 (t, J=11.6 Hz, 3H), 2.70
(dd, J=13.2, 9.5 Hz, 1H), 1.89 (d, J=10.4 Hz, 1H), 1.71 (d, J=13.1
Hz, 3H), 1.43 (dd, J=16.4, 8.5 Hz, 2H), 1.32 (m, 2H), 1.18 (s,
9H).
##STR00248## ##STR00249##
5-thiocyanato-1,3,4-thiadiazol-2-amine (MFH-2-202-1)
[0453] A mixture of 5-bromo-1,3,4-thiadiazol-2-amine (1.5 g, 8.3
mmol) and potassium thiocyanate (3.2 g, 33.3 mmol) in methanol (30
mL) was stirred at room temperature for 20 h. Methanol was
evaporated and water (10 ml) was added. The pH of the aqueous
solution was adjusted to pH=12 with 10% NaOH and precipitate
formed. The solid was collected by filtration to yield compound
MFH-2-202-1 (600 mg, 46%) as solid. LCMS (m/z): 159 [M+H]+.
5-((5-tert-butyloxazol-2-yl)methylthio)-1,3,4-thiadiazol-2-amine
(MFH-2-204-1)
[0454] To a solution of compound MFH-2-202-1 (315 mg, 2 mmol) in
EtOH (5 ml) was added NaBH.sub.4 (151 mg, 4 mmol) at room
temperature. The mixture was stirred for 1h, and then acetone (3
ml) was slowly added. After 1 h, a solution of
5-tert-Butyl-2-chloromethyloxazole (348 mg, 2 mmol) in EtOH (3 ml)
was added. The resulting reaction mixture was heated to reflux for
1h and then was cooled followed by concentration in reduced
pressure. The residue was dissolved in ethyl acetate. The organic
phase was washed with brine (30 mL) and then the solvent was
removed after drying with MgSO.sub.4. The crude product was then
obtained MFH-2-204-1 (310 mg, 57%) as solid LCMS (m/z): 271
[M+H]+.
2-((5-bromo-1,3,4-thiadiazol-2-ylthio)methyl)-5-tert-butyloxazole
(MFH-2-208-1)
[0455] To a solution of CuBr.sub.2 (594 mg, 2.66 mmol) in
acetonitrile (8 mL) at 0.degree. C. was added t-BuONO (275 mg, 2.66
mmol) followed by compound MFH-2-204-1 (600 mg, 2.22 mmol). The
mixture was stirred at 0.degree. C. for 1h and then was warmed up
to room temperature. After stirring for 1 h, ethyl acetate was
added and the organic mixture washed with hydrochloric acid (50
mL), dried over magnesium sulfate, filtered through a pad of silica
gel, and concentrated in reduced pressure. The residue was
chromatographed on silica gel to give the MFH-2-208-1 (700 mg,
94%). LCMS (m/z): 335 [M+H]+.
(R)-tert-butyl3-(5-((5-tert-butyloxazol-2-yl)methylthio)-1,3,4-thiadiazol--
2-ylamino)piperidine-1-carboxylate (MFH-3-2-1)
[0456] The mixture of MFH-2-208-1 (250 mg, 0.75 mmol),
(R)-tert-butyl 3-aminopiperidine-1-carboxylate (225 mg, 1.12 mmol)
and DIEA (242 mg, 1.87 mmol) in NMP (1 mL) was stirred at
140.degree. C. overnight. The residue was extracted with chloroform
and iso-propanol (4:1). The organic phase was washed with brine (50
mL) and dried over Na.sub.2SO.sub.4. After removal of the solvent,
the residue was purified by silica gel (MeOH/DCM=0-20%) to obtain
MFH-3-2-1 (180 mg, yield 53%). LCMS (m/z): 454 [M+H].sup.+.
(R)-5-((5-tert-butyloxazol-2-yl)methylthio)-N-(piperidin-3-yl)-1,3,4-thiad-
iazol-2-amine (MFH-3-11-1)
[0457] To a solution of MFH-3-2-1 (250 mg, 0.55 mmol) in methanol
(5 mL) was added 4N HCl/dioxane (5 mL). The solution was then
stirred for 3h at room temperature and the solvent was removed
under reduced pressure to provide a crude which was directly used
in the next step. LCMS (m/z): 354 [M+H].sup.+.
(R)-(3-(5-((5-tert-butyloxazol-2-yl)methylthio)-1,3,4-thiadiazol-2-ylamino-
)piperidin-1-yl)(4-nitrophenyl)methanone (MFH-3-14-1)
[0458] The mixture of MFH-3-11-1 (140 mg, 0.4 mmol), 4-nitrobenzoyl
chloride (88 mg, 0.48 mmol) in pyridine (3 mL) was stirred
overnight at room temperature. Then the reaction mixture was
concentrated under reduced pressure and the residue was directly
used in the next step. LCMS (m/z): 503 [M+H].sup.+.
(R)-(4-aminophenyl)(3-(5-((5-tert-butyloxazol-2-yl)methylthio)-1,3,4-thiad-
iazol-2-ylamino)piperidin-1-yl)methanone (MFH-3-18-1)
[0459] To a solution of MFH-3-14-1 (200 mg, 0.4 mmol) in ethyl
acetate and methanol (1:1) were added Tin(II) chloride dehydrate
(904 mg, 4 mmol) and conc. HCl (0.2 mL). After stirring for 3 h at
80.degree. C., the reaction mixture was diluted with chloroform and
iso-propanol (4:1), neutralized with saturated NaHCO.sub.3 and
filtered. The filtrate was extracted with chloroform and
iso-propanol (4:1), concentrated under reduced pressure and the
resulting residue was purified by silica gel column chromatography
(MeOH/DCM=0-20%) to give MFH-3-18-1 (45 mg, yield 24%). LCMS (m/z):
473 [M+H]+.
(R)--N-(4-(3-((5-tert-butyloxazol-2-yl)methylthio)-1,3,4-thiadiazol-2-ylam-
ino)piperidine-1-carbonyl)phenyl)acrylamide (MFH-3-25-1)
[0460] To a solution of MFH-3-18-1 (22 mg, 0.05 mmol) and DIPEA
(0.2 mL) in CH.sub.3CN (2 mL) was added acryloyl chloride (6 mg,
0.06 mmol) in DCM (0.2 mL) dropwise. The mixture was then stirred
at 0.degree. C. for 1 h. The solution was then concentrated under
reduced pressure and the residue was purified by prep-HPLC
(MeOH/H.sub.2O, 0.05% TFA) to provide MFH-3-25-1 (8.8 mg, yield
36%). LCMS (m/z): 527 [M+H]+. .sup.1H NMR (500 MHz, DMSO) .delta.
10.29 (s, 1H), 8.02 (d, J=5.0 Hz, 1H), 7.68 (s, 2H), 7.35 (s, 2H),
6.75 (s, 1H), 6.44 (dd, J=17.0, 10.1 Hz, 1H), 6.28 (d, J=16.9 Hz,
1H), 5.86-5.70 (m, 1H), 4.33 (s, 2H), 3.74 (s, 1H), 3.60 (s, 2H),
3.21 (d, J=32.1 Hz, 2H), 1.99 (s, 1H), 1.77 (s, 1H), 1.58 (dd,
J=22.4, 11.7 Hz, 2H), 1.29 (s, 9H).
##STR00250## ##STR00251##
N1-(4-nitrophenyl)cyclohexane-1,3-diamine (MFH-3-26-1)
[0461] The mixture of 1-fluoro-4-nitrobenzene (200 mg, 1.42 mmol),
cyclohexane-1,3-diamine (485 mg, 4.25 mmol) and K.sub.2CO.sub.3
(587 mg, 4.25 mmol) in DMF (3 mL) was stirred at 90.degree. C. for
6h. The solution was then diluted with chloroform and iso-propanol
(4:1). The organic phase was washed with brine (50 mL) and dried
over Na.sub.2SO.sub.4. After removal of the solvent, the residue
was purified by silica gel (MeOH/DCM=0-20%) to obtain MFH-3-26-1
(230 mg, yield 69%). LCMS (m/z): 236 [M+H].sup.+.
(5-((5-tert-butyloxaxzol-2-yl)methylthio)thiazol-2-yl)-N3-(4-nitrophenyl)c-
yclohexane-1,3-diamine (MFH-3-29-1)
[0462] The mixture of MFH-2-83-1 (272 mg, 0.82 mmol), MFH-3-26-1
(230 mg, 0.98 mmol) and DIEA (316 mg, 2.46 mmol) in NMP (2 mL) was
stirred at 140.degree. C. overnight. The solution was diluted with
chloroform and iso-propanol (4:1). The organic phase was washed
with brine (50 mL) and dried over Na.sub.2SO.sub.4. After removal
of the solvent, the residue was purified by silica gel
(MeOH/DCM=0-20%) to obtain MFH-3-29-1 (80 mg, yield 20%). LCMS
(m/z): 458 [M+H].sup.+.
N1-(3
((5-((5-tert-butyloxazol-2-yl)methylthio)thiazol-2-ylamino)cyclohexy-
l)benzene-1,4-diamine (MFH-3-33-1)
[0463] To a solution of MFH-3-29-1 (80 mg, 0.16 mmol) in ethyl
acetate and methanol (1:1) were added Tin(II) chloride dehydrate
(370 mg, 1.6 mmol) and conc. HCl (0.1 mL). After stirring for 3 h
at 80.degree. C., the reaction mixture was diluted with chloroform
and iso-propanol (4:1), neutralized with saturated NaHCO.sub.3 and
filtered. The filtrate was extracted with chloroform and
iso-propanol (4:1), concentrated under reduced pressure and the
resulting residue was purified by silica gel column chromatography
(MeOH/DCM=0-20%) to give MFH-3-33-1 (20 mg, yield 27%). LCMS (m/z):
458 [M+H]+.
N-(4-(3-(5-((5-tert-butyloxazol-2-yl)methylthio)thiazol-2-ylamino)cyclohex-
ylamino)phenyl)acrylamide (MFH-3-35-1)
[0464] To a solution of MFH-3-33-1 (20 mg, 0.04 mmol) and DIPEA
(0.1 mL) in CH.sub.3CN (2 mL) was added acryloyl chloride (4 mg,
0.04 mmol) in DCM (0.2 mL) dropwise. The mixture was then stirred
at -40.degree. C. for 1h. The solution was then concentrated under
reduced pressure and the residue was purified by prep-HPLC
(MeOH/H.sub.2O, 0.05% TFA) to provide MFH-3-35-1 (9.3 mg, yield
41%). LCMS (m/z): 512 [M+H]+.
##STR00252##
(R)-(3-(5-((5-tert-butyloxazol-2-yl)methylthio)thiazol-2-ylamino)piperidi-
n-1-yl)(2-methyl-4-nitrophenyl)methanone (MFH-3-64-1)
[0465] The mixture of MFH-2-85-1 (44 mg, 0.13 mmol),
4-((tert-butoxycarbonylamino)methyl)benzoic acid (47 mg, 0.19
mmol), HOBT (25 mg, 0.19 mmol) and EDCI (36 mg, 0.19 mmol) in DCM
(3 ml) was stirred for overnight. The reaction mixture was diluted
with DCM (25 ml). The organic phase was washed with saturated
Na.sub.2CO.sub.3 and brine (20 mL) and dried over Na.sub.2SO.sub.4.
After removal of the solvent, the residue was purified by silica
gel (MeOH/DCM=0-20%) to obtain MFH-3-64-1 (63 mg, yield 94%). LCMS
(m/z): 516 [M+H]+.
(R)-(4-amino-2-methylphenyl)(3-(5-((5-tert-butyloxazol-2-yl)methylthio)thi-
azol-2-ylamino)piperidin-1-yl)methanone (MFH-3-66-1)
[0466] To a solution of MFH-3-64-1 (63 mg, 0.12 mmol) in ethyl
acetate and methanol (1:1) were added Tin(II) chloride dehydrate
(138 mg, 0.61 mmol) and conc. HCl (0.1 mL). After stirring for 3 h
at 80.degree. C., the reaction mixture was diluted with chloroform
and iso-propanol (4:1), neutralized with saturated NaHCO.sub.3 and
filtered. The filtrate was extracted with chloroform and
iso-propanol (4:1), concentrated under reduced pressure and the
resulting residue was purified by silica gel column chromatography
(MeOH/DCM=0-20%) to give MFH-3-66-1 (20 mg, yield 34%). LCMS (m/z):
486 [M+H]+.
(R)--N-(4-(3-(5-((5-tert-butyloxazol-2-yl)methylthio)thiazol-2-ylamino)pip-
eridine-1-carbonyl)-3-methylphenyl)acrylamide (MFH-3-75-1)
[0467] To a solution of MFH-3-66-1 (20 mg, 0.04 mmol) and DIPEA
(0.1 mL) in CH.sub.3CN (2 mL) was added acryloyl chloride (4 mg,
0.04 mmol) in DCM (0.2 mL) dropwise. The mixture was then stirred
at 0.degree. C. for 1 h. The solution was then concentrated under
reduced pressure and the residue was purified by prep-HPLC
(MeOH/H.sub.2O, 0.05% TFA) to provide MFH-3-75-1 (9.4 mg, yield
41%). LCMS (m/z): 540 [M+H]+. .sup.1H NMR (500 MHz, DMSO) .delta.
10.16 (d, J=35.4 Hz, 1H), 8.03 (d, J=59.1 Hz, 1H), 7.54 (d, J=9.3
Hz, 1H), 7.24-7.08 (m, 1H), 7.00 (d, J=9.0 Hz, 1H), 6.75 (d, J=10.5
Hz, 1H), 6.69 (s, 1H), 6.43 (dt, J=17.0, 10.5 Hz, 1H), 6.32-6.20
(m, 1H), 5.76 (t, J=11.0 Hz, 1H), 3.96 (s, J=11.8 Hz, 2H), 3.59 (d,
J=52.3 Hz, 4H), 3.00 (s, 1H), 2.19 (s, 3H), 1.92 (dd, J=59.5, 33.2
Hz, 2H), 1.57 (d, J=57.5 Hz, 2H), 1.20 (s, 9H).
##STR00253##
(R)-(3-(5-((5-tert-butyloxazol-2-yl)methylthio)thiazol-2-ylamino)piperidi-
n-1-yl)(2-methoxy-4-nitrophenyl)methanone (MFH-3-65-1)
[0468] The mixture of MFH-2-85-1 (44 mg, 0.13 mmol),
4-((tert-butoxycarbonylamino)methyl)benzoic acid (47 mg, 0.19
mmol), HOBT (25 mg, 0.19 mmol) and EDCI (36 mg, 0.19 mmol) in DCM
(3 ml) was stirred overnight. The reaction mixture was diluted with
DCM (25 ml). The organic phase was washed with saturated
Na.sub.2CO.sub.3 and brine (20 mL) and dried over Na.sub.2SO.sub.3.
After removal of the solvent, the residue was purified by silica
gel (MeOH/DCM=0-20%) to obtain MFH-3-64-1 (71 mg, yield 100%). LCMS
(m/z): 532 [M+H]+.
(R)-4-amino-2-methoxyphenyl)(3-(5-((5-tert-butyloxazol-2-yl)methylthio)thi-
azol-2-ylamino)piperidin-1-yl)methanone (MFH-3-67-1)
[0469] To a solution of MFH-3-64-1 (71 mg, 0.13 mmol) in ethyl
acetate and methanol (1:1) were added Tin(II) chloride dehydrate
(151 mg, 0.67 mmol) and conc. HCl (0.1 mL). After stirring for 3 h
at 80.degree. C., the reaction mixture was diluted with chloroform
and iso-propanol (4:1), neutralized with saturated NaHCO.sub.3 and
filtered. The filtrate was extracted with chloroform and
iso-propanol (4:1), concentrated under reduced pressure and the
resulting residue was purified by silica gel column chromatography
(MeOH/DCM=0-20%) to give MFH-3-67-1 (30 mg, yield 45%). LCMS (m/z):
502 [M+H]+.
(R)--N-(4-(3-(5-((5-tert-butyloxazol-2-yl)methylthio)thiazol-2-ylamino)pip-
eridine-1-carbonyl)-3-methoxyphenyl)acrylamide (MFH-3-81-1)
[0470] To a solution of MFH-3-67-1 (30 mg, 0.06 mmol) and DIPEA
(0.1 mL) in CH.sub.3CN (2 mL) was added acryloyl chloride (7 mg,
0.08 mmol) in DCM (0.2 mL) dropwise. The mixture was then stirred
at 0.degree. C. for 1h. The solution was then concentrated under
reduced pressure and the residue was purified by prep-HPLC
(MeOH/H.sub.2O, 0.05% TFA) to provide MFH-3-81-1 (11.8 mg, yield
35%). LCMS (m/z): 556 [M+H]+. .sup.1H NMR (500 MHz, DMSO) .delta.
10.36-10.15 (m, 1H), 8.16-7.83 (m, 1H), 7.49 (d, J=14.3 Hz, 1H),
7.15 (dd, J=19.9, 7.7 Hz, 1H), 7.04-6.94 (m, 1H), 6.74 (dd, J=28.4,
22.6 Hz, 2H), 6.43 (dt, J=18.7, 9.3 Hz, 1H), 6.27 (ddd, J=17.0,
9.5, 1.8 Hz, 1H), 5.83-5.72 (m, 1H), 3.91 (s, 2H), 3.78 (s, 3H),
3.73 (s, 1H), 3.45 (d, J=13.3 Hz, 1H), 3.24 (t, J=32.2 Hz, 1H),
2.88 (dd, J=34.4, 22.2 Hz, 2H), 1.97 (d, J=18.3 Hz, 1H), 1.78 (s,
1H), 1.46 (s, 2H), 1.20 (s, 9H).
##STR00254## ##STR00255##
(1R,3S)--3-(5-((5-tert-butyloxazol-2-yl)methylthio)thiazol-2-ylamino)cycl-
ohexanecarboxylic acid (MFH-3-79-1)
[0471] The mixture of MFH-2-83-1 (200 mg, 0.6 mmol),
3-aminocyclohexanecarboxylic acid (138 mg, 0.96 mmol) and DIEA (233
mg, 1.8 mmol) in NMP (1 mL) was stirred at 140.degree. C. for
overnight. The residue was extracted with chloroform and
iso-propanol (4:1). The organic phase was washed with brine (50 mL)
and dried over Na.sub.2SO.sub.4. After removal of the solvent, the
residue was purified by silica gel (MeOH/DCM=0-20%) to obtain
MFH-3-79-1 (60 mg, yield 26%). LCMS (m/z): 396 [M+H].sup.+.
tert-butyl1-((1R,3S)--3-(5-((5-tert-butyloxazol-2-yl)methylthio)thiazol-2--
ylamino)cyclohexanecarbonyl)piperidin-4-ylcarbamate
(MFH-3-83-1)
[0472] The mixture of MFH-3-79-1 (60 mg, 0.15 mmol), tert-butyl
piperidin-4-ylcarbamate (46 mg, 0.23 mmol), HOBT (31 mg, 0.23 mmol)
and EDCI (44 mg, 0.23 mmol) in DCM (6 ml) was stirred for
overnight. The reaction mixture was diluted with DCM (25 ml). The
organic phase was washed with saturated Na.sub.2CO.sub.3 and brine
(20 mL) and dried over Na.sub.2SO.sub.4. After removal of the
solvent, the residue was purified by silica gel (MeOH/DCM=0-20%) to
obtain MFH-3-83-1 (80 mg, yield 91%). LCMS (m/z): 578 [M+H]+.
(4-aminopiperidin-1-yl)((1R,3S)--3-(5-((5-tert-butyloxazol-2-yl)methylthio-
)triazol-2-ylamino)cyclohexyl)methanone (MFH-3-85-1)
[0473] To a solution of MFH-3-83-1 (80 mg, 0.14 mmol) in methanol
(3 mL) was added 4N HCl/dioxane (3 mL). The solution was then
stirred for 3h at room temperature and the solvent was removed
under reduced pressure to provide a crude which was directly used
in the next step. LCMS (m/z): 478 [M+H].sup.+.
N-(1-((1R,3S)--3-(5-((5-tert-butyloxazol-2-yl)methylthio)thiazol-2-ylamino-
)cyclohexanecarbonyl)piperidin-4-yl)acrylamide (MFH-3-88-1)
[0474] To a solution of MFH-3-85-1 (20 mg, 0.04 mmol) and DIPEA
(0.1 mL) in CH.sub.3CN (2 mL) was added acryloyl chloride (5 mg,
0.05 mmol) in DCM (0.1 mL) dropwise. The mixture was then stirred
at 0.degree. C. for 1h. The solution was then concentrated under
reduced pressure and the residue was purified by prep-HPLC
(MeOH/H.sub.2O, 0.05% TFA) to provide MFH-3-88-1 (3.7 mg, yield
17%). LCMS (m/z): 532 [M+H]+.
##STR00256##
5-((5-tert-butyloxazol-2-yl)methylthio)-N-((1R,3R)-3-(4-nitrophenoxy)cycl-
ohexyl)thiazol-2-amine (MFH-3-97-1)
[0475] The mixture of MFH-2-83-1 (180 mg, 0.54 mmol),
(1R,3R)-3-(4-nitrophenoxy)cyclohexanamine (204 mg, 0.86 mmol) and
DIEA (140 mg, 1.1 mmol) in NMP (1 mL) was stirred at 140.degree. C.
for overnight. The residue was extracted with chloroform and
iso-propanol (4:1). The organic phase was washed with brine (50 mL)
and dried over Na.sub.2SO.sub.4. After removal of the solvent, the
residue was purified by silica gel (MeOH/DCM=0-20%) to obtain
MFH-3-97-1 (80 mg, yield 30%). LCMS (m/z): 489 [M+H].sup.+.
N-((1R,3R)-3-(4-aminophenoxy)cyclohexyl)-5-((5-tert-butyloxazol-2-yl)methy-
lthio)thiazol-2-amine (MFH-3-102-1)
[0476] To a solution of MFH-3-97-1 (80 mg, 0.16 mmol) in ethyl
acetate and methanol (1:1) were added Tin(II) chloride dehydrate
(300 mg, 1.3 mmol) and cone. HCl (0.1 mL). After stirring for 3 h
at 80.degree. C., the reaction mixture was diluted with chloroform
and iso-propanol (4:1), neutralized with saturated NaHCO.sub.3 and
filtered. The filtrate was extracted with chloroform and
iso-propanol(4:1), concentrated under reduced pressure and the
resulting residue was purified by silica gel column chromatography
(MeOH/DCM=0-20%) to give MFH-3-102-1 (40 mg, yield 54%). LCMS
(m/z): 459 [M+H].sup.+.
N-(4-((1R,3R)-3-(5-((5-tert-butyloxazol-2-yl)methylthio)thiazol-2-ylamino)-
cyclohexyloxy)phenyl)acrylamide (MFH-3-103-1)
[0477] To a solution of MFH-3-102-1 (40 mg, 0.09 mmol) and DIPEA
(0.1 mL) in CH.sub.3CN (2 mL) was added acryloyl chloride (10 mg,
0.11 mmol) in DCM (0.2 mL) dropwise. The mixture was then stirred
at 0.degree. C. for 1 h. The solution was then concentrated under
reduced pressure and the residue was purified by prep-HPLC
(MeOH/H.sub.2O, 0.05% TFA) to provide MFH-3-103-1 (9.6 mg, yield
21%). LCMS (m/z): 513 [M+H]+. .sup.1H NMR (500 MHz, DMSO) .delta.
10.00 (s, 1H), 8.04 (d, J=7.3 Hz, 1H), 7.56 (d, J=9.0 Hz, 2H), 6.94
(s, 1H), 6.93-6.89 (m, 2H), 6.71 (s, 1H), 6.39 (dd, J=17.0, 10.1
Hz, 1H), 6.21 (dd, J=17.0, 2.0 Hz, 1H), 5.71 (dd, J=10.1, 2.0 Hz,
1H), 4.66 (s, 1H), 3.94 (s, 2H), 3.83 (s, 1H), 2.01 (d, J=13.3 Hz,
1H), 1.85 (s, 1H), 1.74-1.54 (m, 5H), 1.34 (dd, J=21.1, 11.1 Hz,
1H), 1.18 (s, 9H).
##STR00257## ##STR00258##
(R)-tert-butyl3-(5-cyanothiazol-2-ylamino)piperidine-1-carboxylate
(MFH-3-72-1)
[0478] The mixture of 2-bromothiazole-5-carbonitrile (800 mg, 4.23
mmol), (R)-tert-butyl 3-aminopiperidine-1-carboxylate (1.1 g, 5.5
mmol) and DIEA (820 mg, 6.35 mmol) in THF (15 mL) was stirred at
80.degree. C. for 6h. The residue was extracted with chloroform and
iso-propanol (4:1). The organic phase was washed with brine (50
mL.times.2) and dried over Na.sub.2SO.sub.4. After removal of the
solvent, the residue was purified by silica gel (MeOH/DCM=0-20%) to
obtain MFH-3-72-1 (1.3 g, yield 100%). LCMS (m/z): 309
[M+H].sup.+.
(R)-tert-butyl
3-(5-(aminomethyl)thiazol-2-ylamino)piperidine-1-carboxylate
(MFH-3-82-1)
[0479] To a solution of MFH-3-72-1 (300 mg, 1 mmol) and CoCl.sub.2
6H.sub.2O (232 mg, 1 mmol) in ethanol (8 mL) was added NaBH.sub.4
(110 mg, 3 mmol) at room temperature. The vial was sealed and
stirred at room temperature for 3h and then was quenched with
water. The obtained mixture was extracted with chloroform and
iso-propanol (4:1). The organic phase was washed with brine (50
mL.times.2) and dried over Na.sub.2SO.sub.4. After removal of the
solvent, the residue was purified by silica gel (MeOH/DCM=0-20%) to
obtain MFH-3-82-1 (89 g, yield 28%). LCMS (m/z): 313
[M+H].sup.+.
(R)-tert-butyl3-((5-chloro-4-trifluoromethyl)pyridin-2-ylamino)methyl)thia-
zol-2-ylamino)piperidine-1-carboxylate (MFH-3-99-1)
[0480] The mixture of MFH-2-82-1 (175 mg, 0.56 mmol),
2-bromo-5-chloro-4-(trifluoromethyl)pyridine (146 mg, 0.56 mmol)
and DIEA (108 mg, 0.84 mmol) in NMP (1 mL) was stirred at
80.degree. C. for overnight. The residue was extracted with
chloroform and iso-propanol (4:1). The organic phase was washed
with brine (50 mL.times.2) and dried over Na.sub.2SO.sub.4. After
removal of the solvent, the residue was purified by silica gel
(MeOH/DCM=0-20%) to obtain MFH-3-99-1 (90 mg, yield 58%). LCMS
(m/z): 492 [M+H].sup.+.
(R)-5-((5-chloro-4-(trifluoromethyl)pyridin-2-ylamino)methyl)-N-(piperidin-
-3-yl)thiazol-2-amine (MFH-3-106-1)
[0481] To a solution of MFH-3-99-1 (90 mg, 0.18 mmol) in methanol
(3 mL) was added 4N HCl/dioxane (3 mL). The solution was then
stirred for 3h at room temperature and the solvent was removed
under reduced pressure to provide a crude which was directly used
in the next step. LCMS (m/z): 392 [M+H].sup.+.
(R)--N-(4-(3-(5-((5-chloro-4-trifluoromethyl)pyridin-2-ylamino)methyl)thia-
zol-2-ylamino)piperidine-1-carbonyl)phenyl)acrylamide
(MFH-3-107-1)
[0482] The mixture of MFH-3-106-1 (20 mg, 0.05 mmol),
4-acrylamidobenzoic acid (10 mg, 0.05 mmol), HOBT (8 mg, 0.06 mmol)
and EDCI (11 mg, 0.06 mmol) in DMF (0.5 ml). Then the reaction
mixture was stirred for overnight. The reaction mixture was
purified by prep-HPLC (MeOH/H.sub.2O, 0.05% TFA) to provide
MFH-3-107-1 (3.8 mg, yield 13%). LCMS (m/z): 565 [M+H]+. .sup.1H
NMR (500 MHz, DMSO) .delta. 10.31 (s, 1H), 8.28 (s, 1H), 7.68 (s,
3H), 7.36 (d, J=7.2 Hz, 2H), 7.25-6.81 (m, 3H), 6.45 (dd, J=17.0,
10.2 Hz, 1H), 6.28 (dd, J=17.0, 1.9 Hz, 1H), 5.78 (dd, J=10.1, 1.9
Hz, 1H), 4.46 (s, 2H), 3.69 (s, 3H), 3.18 (d, J=11.7 Hz, 2H), 1.96
(s, 1H), 1.75 (s, 1H), 1.55 (m, 2H).
##STR00259## ##STR00260##
diethyl (5-tert-butyloxazol-2-yl)methylphosphonate (MFH-3-91-1)
[0483] A solution of 5-tert-butyl-2-(chloromethyl)oxazole (800 mg,
4.61 mmol) in triethyl phosphite (3.83 g, 23.04 mmol) was heated at
120.degree. C. for overnight. The solvent was removed under reduced
pressure to provide a crude which was directly used in the next
step. LCMS (m/z): 276 [M+H].sup.+.
(E)-tert-butyl5-(2-(5-tert-butyloxazol-2-yl)vinyl)thiazol-2-ylcarbamate
(MFH-3-94-1)
[0484] To a solution of MFH-3-91-1 (500 mg, 1.82 mmol) in THF (5
mL) was added t-BuOK (370 mg, 3.3 mmol) at room temperature. The
vial was sealed and stirred at room temperature for 10 min. Then, a
solution of tert-butyl 5-formylthiazol-2-ylcarbamate (345 mg, 1.5
mmol) in THF (3 mL) was added. After completion, the reaction
mixture was quenched with water. The obtained mixture was extracted
with chloroform and iso-propanol (4:1). The organic phase was
washed with brine (50 mL.times.2) and dried over Na.sub.2SO.sub.4.
After removal of the solvent, the residue was purified by silica
gel (MeOH/DCM=0-20%) to obtain MFH-3-94-1 (440 mg, yield 84%). LCMS
(m/z): 350 [M+H].sup.+.
(E)-5-(2-(5-tert-butyloxzol-2-yl)vinyl)thiazol-2-amine
(MFH-3-98-1)
[0485] To a solution of MFH-3-94-1 (220 mg, 0.63 mmol) in methanol
(5 mL) was added 4N HCl/dioxane (5 mL). The solution was then
stirred for 3h at room temperature and the solvent was removed
under reduced pressure to provide a crude which was directly used
in the next step. LCMS (m/z): 250 [M+H].sup.+.
(E)-2-(2-(2-bromothiazol-5-yl)vinyl)-5-tert-butyloxazole
(MFH-3-105-1)
[0486] To a solution of CuBr.sub.2 (172 mg, 0.77 mmol) in
acetonitrile (15 mL) at 0.degree. C. was added t-BuONO (79 mg, 0.77
mmol) followed by compound MFH-3-98-1 (160 mg, 0.64 mmol). The
mixture was stirred at 0.degree. C. for one hour, then at room
temperature for one hour, ethyl acetate was added and the organic
mixture washed with hydrochloric acid (20 mL), dried over magnesium
sulfate, filtered through a pad of silica gel, and concentrated
under reduced pressure. The residue was chromatographed on silica
gel to give the MFH-3-105-1 (50 mg, 25%). LCMS (m/z): 314
[M+H]+.
(R,E)-tert-butyl3-(5-(2-(5-tert-butyloxazol-2-yl))vinyl)thiazol-2-ylamino)-
piperidine-1-carboxylate (MFH-3-108-1)
[0487] The mixture of MFH-3-105-1 (50 mg, 0.16 mmol),
(R)-tert-butyl 3-aminopiperidine-1-carboxylate (51 mg, 0.26 mmol)
and DIEA (41 mg, 0.32 mmol) in NMP (0.5 mL) was stirred at
140.degree. C. for overnight. The residue was extracted with
chloroform and iso-propanol (4:1). The organic phase was washed
with brine (10 mL.times.2) and dried over Na.sub.2SO.sub.4. After
removal of the solvent, the residue was purified by silica gel
(MeOH/DCM=0-20%) to obtain MFH-3-108-1 (40 mg, yield 58%). LCMS
(m/z): 433 [M+H].sup.+.
(R,E)-5-(2-(5-tert-butyloxazol-2-yl)vinyl)-N-(piperidin-3-yl)thiazol-2-ami-
ne (MFH-3-109-1)
[0488] To a solution of MFH-3-108-1 (40 g, 0.09 mmol) in methanol
(2 mL) was added 4N HCl/dioxane (2 mL). The solution was then
stirred for 3h at room temperature and the solvent was removed
under reduced pressure to provide a crude which was directly used
in the next step. LCMS (m/z): 333 [M+H].sup.+.
(R,E)-N-(4-(3-(5-(2-(5-tert-butyloxazol-2-yl)vinyl)triazol-2-ylamino)piper-
idine-1-carbonyl)phenyl)acrylamide (MFH-3-110-1)
[0489] The mixture of MFH-3-109-1 (35 mg, 0.1 mmol),
4-acrylamidobenzoic acid (30 mg, 0.16 mmol), HOBT (21 mg, 0.16
mmol) and EDCI (30 mg, 0.16 mmol) in DMF (0.5 ml). Then the
reaction mixture was stirred for overnight. The reaction mixture
was purified by prep-HPLC (MeOH/H.sub.2O, 0.05% TFA) to provide
MFH-3-110-1 (5 mg, yield 10%). LCMS (m/z): 506 [M+H]+.
##STR00261## ##STR00262##
5-(2-(5-tert-butyloxazol-2-yl)ethyl)thiazol-2-amine
(MFH-3-104-1)
[0490] A mixture of compound MFH-3-94-1 (250 mg, 1 mmol) and 10%
Pd/C (20 mg) in MeOH (10 mL) was stirred for 5h at room temperature
under H.sub.2 balloon. The mixture was filtered through celite. The
filtrate was added 4N HCl/dioxane (5 mL). The solution was then
stirred for 3h at room temperature and the solvent was removed
under reduced pressure to provide a crude which was directly used
in the next step. LCMS (m/z): 252 [M+H].sup.+.
2-(2-(2-bromothiazol-5-yl)ethyl)-5-tert-butyloxazole
(MFH-3-114-1)
[0491] To a solution of CuBr.sub.2 (53 mg, 0.24 mmol) in
acetonitrile (2 mL) at 0.degree. C. was added t-BuONO (25 mg, 0.24
mmol) followed by compound MFH-3-104-1 (60 mg, 0.24 mmol). The
mixture was stirred at 0.degree. C. for one hour, and then was
warmed up to room temperature. Ethyl acetate was added and the
organic mixture was washed with hydrochloric acid (20 mL), dried
over magnesium sulfate, filtered through a pad of silica gel, and
concentrated in vacuo. The residue was chromatographed on silica
gel to give the MFH-3-114-1 (40 mg, 53%). LCMS (m/z): 316
[M+H]+.
(R)-tert-butyl3-(5-(2-(5-tert-butyloxazol-2-yl)ethyl)thiazol-2-ylamino)pip-
eridin-1-carboxylate (MFH-3-115-1)
[0492] The mixture of MFH-3-114-1 (40 mg, 0.13 mmol),
(R)-tert-butyl 3-aminopiperidine-1-carboxylate (41 mg, 0.2 mmol)
and DIEA (33 mg, 0.25 mmol) in NMP (0.5 mL) was stirred at
140.degree. C. for overnight. The residue was extracted with
chloroform and iso-propanol (4:1). The organic phase was washed
with brine (10 mL.times.2) and dried over Na.sub.2SO.sub.4. After
removal of the solvent, the residue was purified by silica gel
(MeOH/DCM=0-20%) to obtain MFH-3-115-1 (35 mg, yield 63%). LCMS
(m/z): 435 [M+H].sup.+.
(R)-5-(2-(5-tert-butyloxazol-2-yl)ethyl)-N-(piperidin-3-yl)thiazol-2-amine
[0493] To a solution of MFH-3-115-1 (35 mg, 0.08 mmol) in methanol
(2 mL) was added 4N HCl/dioxane (2 mL). The solution was then
stirred for 3h at room temperature and the solvent was removed
under reduced pressure to provide a crude which was directly used
in the next step. LCMS (m/z): 335 [M+H].sup.+.
(R)--N-(4-(3-(5-(2-(5-tert-butyloxazol-2-yl)ethyl)thiazol-2-ylamino)piperi-
dine-1-carbonyl)phenyl)acrylamide (MFH-3-116-1)
[0494] The mixture of
(R)-5-(2-(5-tert-butyloxazol-2-yl)ethyl)-N-(piperidin-3-yl)thiazol-2-amin-
e (23 mg, 0.07 mmol), 4-acrylamidobenzoic acid (15 mg, 0.08 mmol),
HOBT (12 mg, 0.09 mmol) and EDCI (17 mg, 0.09 mmol) in DMF (0.5 ml)
was stirred for overnight. The reaction mixture was purified by
prep-HPLC (MeOH/H.sub.2O, 0.05% TFA) to provide MFH-3-116-1 (5.5
mg, yield 16%). LCMS (m/z): 508 [M+H]+. .sup.1H NMR (500 MHz, DMSO)
.delta. 10.38 (s, 1H), 8.29 (d, J=7.5 Hz, 1H), 7.82 (d, J=8.8 Hz,
2H), 7.74 (d, J=8.8 Hz, 2H), 6.96 (s, 1H), 6.68 (s, 1H), 6.45 (dd,
J=17.0, 10.2 Hz, 1H), 6.29 (dd, J=17.0, 1.9 Hz, 1H), 5.79 (dd,
J=10.1, 1.9 Hz, 1H), 3.87 (dd, J=12.3, 4.0 Hz, 1H), 3.75-3.63 (m,
2H), 3.13-2.99 (m, 4H), 2.96 (t, J=6.8 Hz, 2H), 1.88 (m, 2H), 1.63
(m, 2H), 1.22 (s, 9H).
##STR00263## ##STR00264##
2-bromo-5-((3-(trifluoromethyl)benzyloxy)methyl)thiazole
(MFH-3-111-1)
[0495] The mixture of (2-bromothiazol-5-yl)methanol (180 mg, 0.93
mmol), 1-(bromomethyl)-3-(trifluoromethyl)benzene (288 mg, 1.2
mmol) and NaOH (67 mg, 1.67 mmol) in DMF (2 mL) was stirred at
120.degree. C. for 3h. The residue was extracted with chloroform
and iso-propanol (4:1). The organic phase was washed with brine (20
mL.times.2) and dried over Na.sub.2SO.sub.4. After removal of the
solvent, the residue was purified by silica gel (PE/EA=0-50%) to
obtain MFH-3-111-1 (160 mg, yield 49%). LCMS (m/z): 353
[M+H].sup.+.
(R)-tert-butyl35-((3-(trifluoromethyl)benzyloxy)methyl)thiazol-2-ylamino)p-
iperidine-1-carboxylate (MFH-3-112-1)
[0496] The mixture of MFH-3-111-1 (160 mg, 0.34 mmol),
(R)-tert-butyl 3-aminopiperidine-1-carboxylate (109 mg, 0.55 mmol)
and DIEA (88 mg, 0.68 mmol) in NMP (1 mL) was stirred at
140.degree. C. for overnight. The residue was extracted with
chloroform and iso-propanol (4:1). The organic phase was washed
with brine (20 mL.times.2) and dried over Na.sub.2SO.sub.4. After
removal of the solvent, the residue was purified by silica gel
(MeOH/DCM=0-20%) to obtain MFH-3-112-1 (35 mg, yield 22%). LCMS
(m/z): 472 [M+H].sup.+.
(R)--N-(piperidin-3-yl)-5-((3-(trifluoromethyl)benzyloxy)methyl)thiazol-2--
amine (MFH-3-119-1)
[0497] To a solution of MFH-3-112-1 (35 mg, 0.08 mmol) in methanol
(2 mL) was added 4N HCl/dioxane (2 mL). The solution was then
stirred for 3h at room temperature and the solvent was removed
under reduced pressure to provide a crude which was directly used
in the next step. LCMS (m/z): 372 [M+H].sup.+.
(R)--N-(4-(3-(5-((3-(trifluoromethyl)benzyloxy)methyl)thiazol-2-ylamino)pi-
peridine-1-carbonyl)phenyl)acrylamide (MFH-3-120-1)
[0498] The mixture of MFH-3-119-1 (23 mg, 0.06 mmol),
4-acrylamidobenzoic acid (15 mg, 0.08 mmol), HOBT (12 mg, 0.09
mmol) and EDCI (17 mg, 0.09 mmol) in DMF (0.5 ml). Then the
reaction mixture was stirred for overnight. The reaction mixture
was purified by prep-HPLC (MeOH/H.sub.2O, 0.05% TFA) to provide
MFH-3-120-1 (8.7 mg, yield 26%). LCMS (m/z): 545 [M+H]+. .sup.1H
NMR (500 MHz, DMSO) .delta. 10.32 (s, 1H), 7.72-7.57 (m, 7H), 7.37
(d, J=5.1 Hz, 3H), 6.47-6.40 (m, 1H), 6.27 (dd, J=16.6, 6.9 Hz,
1H), 5.77 (t, J=8.3 Hz, 1H), 4.58 (d, J=8.2 Hz, 2H), 4.56 (s, 2H),
3.78 (s, 3H), 3.20 (d, J=6.7 Hz, 2H), 1.98 (s, 1H), 1.76 (s, 1H),
1.55 (m, 2H).
##STR00265## ##STR00266##
2-(((2-bromothiazol-5-yl)methoxy)methyl)-5-tert-butyloxazole
(MFH-3-121-1)
[0499] The mixture of (2-bromothiazol-5-yl)methanol (220 mg, 1.1
mmol), 5-tert-butyl-2-(chloromethyl)oxazole (256 mg, 1.5 mmol) and
NaOH (82 mg, 2.04 mmol) in DMF (2 mL) was stirred at 120.degree. C.
for 3h. The residue was extracted with chloroform and iso-propanol
(4:1). The organic phase was washed with brine (20 mL) and dried
over Na.sub.2SO.sub.4. After removal of the solvent, the residue
was purified by silica gel (PE/EA=0-50%) to obtain MFH-3-111-1 (170
mg, yield 45%). LCMS (m/z): 332 [M+H].sup.+.
(R)-tert-butyl3-(5-(((5-tert-butyloxazol-2-yl)methoxy)methyl)triazol-2-yla-
mino)piperidine-1-carboxylate (MFH-3-123-1)
[0500] The mixture of MFH-3-121-1 (170 mg, 0.51 mmol),
(R)-tert-butyl 3-aminopiperidine-1-carboxylate (164 mg, 0.82 mmol)
and DIEA (133 mg, 1 mmol) in NMP (1 mL) was stirred at 140.degree.
C. for overnight. The residue was extracted with chloroform and
iso-propanol (4:1). The organic phase was washed with brine (20 mL)
and dried over Na.sub.2SO.sub.4. After removal of the solvent, the
residue was purified by silica gel (MeOH/DCM=0-20%) to obtain
MFH-3-123-1 (50 mg, yield 22%). LCMS (m/z): 451 [M+H].sup.+.
(R)-5-(((5-tert-butyloxazol-2-yl)methoxy)methyl)-N-(piperidin-3-yl)thiazol-
-2-amine (MFH-3-126-1)
[0501] To a solution of MFH-3-123-1 (50 mg, 0.11 mmol) in methanol
(2 mL) was added 4N HCl/dioxane (2 mL). The solution was then
stirred for 3h at room temperature and the solvent was removed
under reduced pressure to provide a crude which was directly used
in the next step. LCMS (m/z): 372 [M+H].sup.+.
(R)--N-(4-(3-(((5-tert-butyloxazol-2-yl)methoxy)methyl)thiazol-2-ylamino)p-
iperidine-1-carbonyl)phenyl)acrylamide (MFH-3-128-1)
[0502] The mixture of MFH-3-126-1 (23 mg, 0.06 mmol),
4-acrylamidobenzoic acid (15 mg, 0.08 mmol), HOBT (12 mg, 0.09
mmol) and EDCI (17 mg, 0.09 mmol) in DMF (0.5 ml) was stirred for
overnight. The reaction mixture was purified by prep-HPLC
(MeOH/H.sub.2O, 0.05% TFA) to provide MFH-3-128-1 (7 mg, yield
22%). LCMS (m/z): 524 [M+H]+. .sup.1H NMR (500 MHz, DMSO) .delta.
10.37 (s, 1H), 8.33 (d, J=38.9 Hz, 1H), 7.84 (d, J=8.5 Hz, 2H),
7.75 (d, J=8.4 Hz, 2H), 7.37 (d, J=8.3 Hz, 1H), 6.89-6.71 (m, 1H),
6.46 (dd, J=16.9, 10.1 Hz, 1H), 6.29 (d, J=17.0 Hz, 1H), 5.80 (d,
J=10.0 Hz, 1H), 4.59-4.40 (m, 2H), 4.05 (s, 1H), 3.99-3.85 (m, 2H),
3.33 (dd, J=31.2, 10.4 Hz, 2H), 3.00 (m, 2H), 1.93 (d, J=15.1 Hz,
1H), 1.84 (s, 1H), 1.62 (s, 2H), 1.23 (s, 9H).
##STR00267## ##STR00268##
(R)-tert-butyl3-(5-(((5-tert-butyloxazol-2-yl)methylamino)methyl)thiazol--
2-ylamino)piperidine-1-carboxylate (MFH-3-90-1)
[0503] The mixture of MFH-2-82-1 (160 mg, 0.51 mmol),
5-tert-butyl-2-(chloromethyl)oxazole (89 mg, 0.51 mmol) and DIEA
(132 mg, 1 mmol) in NMP (1 mL) was stirred at 80.degree. C. for
overnight. The solution was extracted with chloroform and
iso-propanol (4:1). The organic phase was washed with brine (50
mL.times.2) and dried over Na.sub.2SO.sub.4. After removal of the
solvent, the residue was purified by silica gel (MeOH/DCM=0-20%) to
obtain MFH-3-90-1 (133 mg, yield 58%). LCMS (m/z): 450
[M+H].sup.+.
(R)-5-(((5-tert-butyloxazol-2-yl)methylamino)methyl)-N-(piperidin-3-yl)thi-
azol-2-amine (MFH-3-136-1)
[0504] To a solution of MFH-3-90-1 (60 mg, 0.13 mmol) in methanol
(3 mL) was added 4N HCl/dioxane (3 mL). The solution was then
stirred for 3h at room temperature and the solvent was removed
under reduced pressure to provide a crude which was directly used
in the next step. LCMS (m/z): 350 [M+H]+.sup.+.
(R)--N-(4-(3-(5-((5-tert-butyloxazol-2-yl)methylamino)methyl)thiazol-2-yla-
mino)piperidine-1-carbonyl)phenyl)acrylamide (MFH-3-137-1)
[0505] The mixture of MFH-3-136-1 (30 mg, 0.09 mmol),
4-acrylamidobenzoic acid (17 mg, 0.09 mmol) and TEA (9 mg, 0.09
mmol) in DMF (1 ml). The reaction mixture was cooled to -60.degree.
C., and HATU (33 mg, 0.09 mmol) was added in the reaction mixture.
The reaction mixture was purified by prep-HPLC (MeOH/H.sub.2O,
0.05% TFA) to provide MFH-3-137-1 (18 mg, yield 38%). LCMS (m/z):
523 [M+H]+. .sup.1H NMR (500 MHz, DMSO) .delta. 10.31 (s, 1H),
8.79-8.59 (m, 2H), 8.10-7.97 (m, 1H), 7.69 (s, 1H), 7.37 (s, 1H),
7.13 (d, J=6.5 Hz, 1H), 6.96 (dd, J=9.8, 4.5 Hz, 1H), 6.78 (s, 1H),
6.43 (d, J=6.7 Hz, 1H), 6.28 (s, 1H), 5.79 (dd, J=12.2, 4.2 Hz,
1H), 4.31 (s, 2H), 3.86 (s, 2H), 3.72 (s, 2H), 3.43 (d, J=10.6 Hz,
1H), 2.93-2.75 (m, 2H), 2.00 (d, J=8.6 Hz, 1H), 1.89 (d, J=22.0 Hz,
1H), 1.59 (m, 2H), 1.23 (s, 9H).
##STR00269##
(R)-tert-butyl4-(3-(5-((5-tert-butyloxazol-2-yl)methylthio)triazol-2-ylam-
ino)piperidin-1-ylsulfonyl)phenylcarbamate (MFH-3-147-1)
[0506] The mixture of MFH-2-85-1 (60 mg, 0.17 mmol) and TEA (26 mg,
0.26 mmol) in DCM (1 ml) was cooled to 0.degree. C. and then a
solution of tert-butyl 4-(chlorosulfonyl)phenylcarbamate (50 mg,
0.17 mmol) in DCM (1 mL) was added. The reaction mixture was
extracted with chloroform and iso-propanol (4:1). The organic phase
was washed with brine (50 mL) and dried over Na.sub.2SO.sub.4.
After removal of the solvent, the residue was purified by silica
gel (MeOH/DCM=0-20%) to obtain MFH-3-147-1 (80 mg, yield 78%). LCMS
(m/z): 608 [M+H].sup.+.
(R)--N-(1-(4-aminophenylsulfonyl)piperidin-3-yl)-5-((5-tert-butyloxazol-2--
yl)methylthio)thiazol-2-amine (MFH-3-148-1)
[0507] To a solution of MFH-3-147-1 (80 mg, 0.13 mmol) in methanol
(3 mL) was added 4N HCl/dioxane (3 mL). The solution was then
stirred for 3h at room temperature and the solvent was removed
under reduced pressure to provide a crude which was directly used
in the next step. LCMS (m/z): 507 [M+H].sup.+.
(R)--N-(4-(3-(5-((5-tert-butyloxazol-2-yl)methylthio)thiazol-2-ylamino)pip-
eridin-1-ylsulfonyl)phenyl)acrylamide (MFH-3-151-1)
[0508] To a solution of MFH-3-148-1 (30 mg, 0.06 mmol) and DIPEA
(0.2 mL) in CH.sub.3CN (2 mL) was added acryloyl chloride (7 mg,
0.08 mmol) in DCM (0.2 mL) dropwise. The mixture was then stirred
at 0.degree. C. for 1 h. The solution was then concentrated under
reduced pressure and the residue was purified by prep-HPLC
(MeOH/H.sub.2O, 0.05% TFA) to provide MFH-3-151-1 (8.5 mg, yield
25%). LCMS (m/z): 561 [M+H]+. .sup.1H NMR (500 MHz, DMSO) .delta.
10.56 (d, J=22.3 Hz, 1H), 8.00 (d, J=7.3 Hz, 1H), 7.93-7.89 (m,
1H), 7.87 (dd, J=11.3, 4.4 Hz, 1H), 7.83-7.75 (m, 1H), 7.72-7.68
(m, 1H), 6.98 (d, J=2.7 Hz, 1H), 6.75-6.70 (m, 1H), 6.51-6.42 (m,
1H), 6.32 (ddd, J=17.0, 3.9, 1.9 Hz, 1H), 5.83 (ddd, J=10.1, 6.1,
1.9 Hz, 1H), 3.96 (d, J=3.8 Hz, 2H), 3.43-3.34 (m, 1H), 3.25 (d,
J=11.8 Hz, 1H), 3.07-2.93 (m, 1H), 2.86 (dd, J=12.6, 9.7 Hz, 1H),
2.32-2.22 (m, 1H), 1.77 (d, J=6.7 Hz, 1H), 1.67 (d, J=3.5 Hz, 1H),
1.52 (d, J=10.2 Hz, 1H), 1.34 (dd, J=18.1, 10.1 Hz, 1H), 1.24-1.16
(m, 9H).
##STR00270## ##STR00271##
tert-butyl 5-(benzylthio)indoline-1-carboxylate (MFH-3-158-1)
[0509] A mixture of tert-butyl 5-bromoindoline-1-carboxylate (600
mg, 2.02 mmol), phenylmethanethiol (276 mg, 2.22 mmol),
Pd.sub.2(dba).sub.3 (185 mg, 0.2 mmol), Xantphos (117 mg, 0.2
mmmol), and DIEA (783 mg, 6.06 mmol) in toluene (25 mL) was
refluxed under N.sub.2 atmosphere for 5h. After cooling to room
temperature, ethyl acetate (50 mL) and H.sub.2O (50 mL) were added
to the mixture, and insoluble materials were removed by filtration.
The filtrate was diluted with ethyl acetate (50 mL) and was washed
with water, 0.2 M hydrochloric acid, aqueous NaHCO.sub.3 solution,
and brine. The organic layer was dried over Na.sub.2SO.sub.4 and
concentrated under reduced pressure. The residue was crystallized
from AcOEt-i-Pr.sub.2O to give the title compound as a yellow solid
(660 mg, 96%). LCMS (m/z): 342 [M+H]+.
tert-butyl 5-(chlorosulfonyl)indoline-1-carboxylate
(MFH-3-159-1)
[0510] To a mixture of compound MFH-3-158-1 (660 g, 1.94 mmol),
acetic acid (6 mL) in water (3 mL) was added N-chlorosuccinimide
(1.1 g, 7.74 mol) at 0.degree. C. The mixture was stirred at room
temperature for 4 h. Precipitated solid was collected by
filtration, washed with water and dried to give the title compound
as a pale yellow solid (400 mg, 65%). This compound was used for
the next step without further purification.
(R)-tert-butyl5-(3-(5-((5-tert-butyloxazol-2-yl)methylthio)thiazol-2-ylami-
no)piperidin-1-ylsulfonyl)indoline-1-carboxylate (MFH-3-162-1)
[0511] The mixture of MFH-2-85-1 (60 mg, 0.17 mmol) and TEA (26 mg,
0.26 mmol) in DCM (1 ml) was cooled to 0.degree. C. and a solution
of MFH-3-159-1 (54 mg, 0.17 mmol) in DCM (1 mL) was added. And
reaction completed, the reaction mixture was extracted with
chloroform and iso-propanol (4:1). The organic phase was washed
with brine (50 mL) and dried over Na.sub.2SO.sub.4. After removal
of the solvent, the residue was purified by silica gel
(MeOH/DCM=0-20%) to obtain MFH-3-162-1 (82 mg, yield 76%). LCMS
(m/z): 634 [M+H].sup.+.
(R)-5-((5-tert-butyloxazol-2-yl)methylthio)-N-(1-(indolin-5-ylsulfonyl)pip-
eridin-3-yl)thiazol-2-amine (MFH-3-164-1)
[0512] To a solution of MFH-3-162-1 (82 mg, 0.13 mmol) in methanol
(3 mL) was added 4N HCl/dioxane (3 mL). The solution was then
stirred for 3h at room temperature and the solvent was removed
under reduced pressure to provide a crude which was directly used
in the next step. LCMS (m/z): 534 [M+H].sup.+.
(R)-1-(5-(3-(5-((5-tert-butyloxazol-2-yl)methylthio)thiazol-2-ylamino)pipe-
ridin-1-ylsulfonyl)indolin-1-yl)prop-2-en-1-one (MFH-3-168-1)
[0513] To a solution of MFH-3-164-1 (20 mg, 0.04 mmol) and DIPEA
(0.1 mL) in CH.sub.3CN (2 mL) was added acryloyl chloride (5 mg,
0.05 mmol) in DCM (0.1 mL) dropwise. The mixture was then stirred
at 0.degree. C. for 1h. The solution was then concentrated under
reduced pressure and the residue was purified by prep-HPLC
(MeOH/H.sub.2O, 0.05% TFA) to provide MFH-3-168-1 (5 mg, yield
23%). LCMS (m/z): 588 [M+H]+. .sup.1H NMR (500 MHz, DMSO) .delta.
8.29 (s, 1H), 7.94 (d, J=7.4 Hz, 1H), 7.57 (dd, J=12.2, 3.8 Hz,
2H), 6.96 (s, 1H), 6.81-6.74 (m, 1H), 6.74 (s, 1H), 6.35 (dd,
J=16.7, 2.0 Hz, 1H), 5.89 (dd, J=10.3, 2.0 Hz, 1H), 4.29 (t, J=8.7
Hz, 2H), 3.96 (s, 2H), 3.69 (s, 1H), 3.55 (d, J=8.0 Hz, 1H), 3.25
(t, J=8.4 Hz, 3H), 2.31-2.24 (m, 1H), 2.18 (t, J=7.4 Hz, 1H), 1.77
(d, J=6.6 Hz, 2H), 1.49 (dd, J=17.4, 10.0 Hz, 2H), 1.24 (s,
9H).
##STR00272## ##STR00273##
5-(pyridin-4-ylmethylthio)thiazol-2-amine (MFH-3-146-1)
[0514] To a solution of compound MFH-2-76-1 (300 mg, 1.91 mmol) in
EtOH (5 ml) was added NaBH.sub.4 (144 mg, 3.82 mmol) room
temperature. The mixture was stirred for 1h, and then acetone (3
ml) was slowly introduced. After 1 h, a solution of
4-(bromomethyl)pyridine hydrobromide (483 mg, 1.91 mmol) in EtOH (3
ml) was added. The resulting dark reaction mixture was heated to
reflux for 1h, and was then cooled and concentrated in vacuo. The
residue was partitioned between EtOAc and brine. The organic phase
was separated, dried (MgSO.sub.4), and concentrated in vacuo to
give a crude solid which was triturated with diethyl ether/hexane
to provide compound MFH-3-146-1 (240 mg, 56%) as solid LCMS (m/z):
224 [M+H]+.
2-bromo-5-(pyridin-4-ylmethylthio)thiazole (MFH-3-153-1)
[0515] To a solution of CuBr.sub.2 (288 mg, 1.29 mmol) in
acetonitrile (8 mL) at 0.degree. C. was added t-BuONO (133 mg, 1.29
mmol) followed by compound MFH-3-146-1 (240 g, 1.08 mmol). The
mixture was stirred at 0.degree. C. for one hour, then at room
temperature for one hour. Ethyl acetate was added and the organic
mixture washed with hydrochloric acid (20 mL), dried over magnesium
sulfate, filtered through a pad of silica gel, and concentrated in
vacuo. The residue was chromatographed on silica gel to give the
MFH-3-153-1 (130 mg, 42%). LCMS (m/z): 288 [M+H]+.
(R)-tert-butyl3-(5-(pyridin-4-ylmethylthio)thiazol-2-ylamino)piperidine-1--
carboxylate (MFH-3-156-1)
[0516] The mixture of MFH-3-153-1 (130 mg, 0.45 mmol),
(R)-tert-butyl 3-aminopiperidine-1-carboxylate (145 mg, 0.72 mmol)
and DIEA (117 mg, 0.91 mmol) in NMP (1 mL) was stirred at
140.degree. C. for overnight. The residue was extracted with
chloroform and iso-propanol (4:1). The organic phase was washed
with brine (20 mL) and dried over Na.sub.2SO.sub.4. After removal
of the solvent, the residue was purified by silica gel
(MeOH/DCM=0-20%) to obtain MFH-3-156-1 (90 mg, yield 49%). LCMS
(m/z): 407 [M+H].sup.+.
(R)--N-(piperidin-3-yl)-5-(pyridin-4-ylmethylthio)thiazol-2-amine
(MFH-3-157-1)
[0517] To a solution of MFH-3-156-1 (90 mg, 0.22 mmol) in methanol
(2 mL) was added 4N HCl/dioxane (2 mL). The solution was then
stirred for 3h at room temperature and the solvent was removed
under reduced pressure to provide a crude which was directly used
in the next step. LCMS (m/z): 307 [M+H].sup.+.
(R)-tert-butyl4-(3-(5-(pyridin-4-ylmethylthio)thiazol-2-ylamino)piperidine-
-1-carbonyl)phenylcarbamate (MFH-3-177-1)
[0518] The mixture of MFH-3-157-1 (60 mg, 0.2 mmol),
4-(tert-butoxycarbonylamino)benzoic acid (15 mg, 0.08 mmol), HOBT
(12 mg, 0.09 mmol) and EDCI (60 mg, 0.26 mmol) in DMF (1 ml) was
stirred for overnight. The reaction mixture was diluted with
chloroform and iso-propanol (4:1). The saturated Na.sub.2CO.sub.3
was added and the reaction mixture was extracted with chloroform
and iso-propanol (4:1), concentrated under reduced pressure and the
resulting residue was purified by silica gel column chromatography
(MeOH/DCM=0-20%) to give MFH-3-177-1 (90 mg, yield 86%). LCMS
(m/z): 526 [M+H]+.
(R)-(4-aminophenyl)(3-(5-(pyridin-4-ylmethylthio)thiazol-2-ylamino)piperid-
in-1-yl)methanone (MFH-3-178-1)
[0519] To a solution of MFH-3-177-1 (90 mg, 0.17 mmol) in methanol
(2 mL) was added 4N HCl/dioxane (2 mL). The solution was then
stirred for 3h at room temperature and the solvent was removed
under reduced pressure to provide a crude which was directly used
in the next step. LCMS (m/z): 426 [M+H].sup.+.
(R)--N-(4-(3-(5-(pyridin-4-ylmethylthio)thiazol-2-ylamino)piperidine-1-car-
bonyl)phenyl)acrylamide (MFH-3-179-1)
[0520] To a solution of MFH-3-178-1 (40 mg, 0.09 mmol) and DIPEA
(0.2 mL) in CH.sub.3CN (2 mL) was added acryloyl chloride (11 mg,
0.12 mmol) in DCM (0.1 mL) dropwise. The mixture was then stirred
at 0.degree. C. for 1h. The solution was then concentrated under
reduced pressure and the residue was purified by prep-HPLC
(MeOH/H.sub.2O, 0.05% TFA) to provide MFH-3-179-1 (4.4 mg, yield
10%). LCMS (m/z): 480 [M+H]+. .sup.1H NMR (500 MHz, DMSO) .delta.
10.30 (s, 1H), 8.06 (s, 1H), 7.85 (s, 2H), 7.68 (s, 2H), 7.36 (s,
2H), 7.25-7.02 (m, 2H), 6.80 (s, 1H), 6.49-6.37 (m, 1H), 6.27 (d,
J=16.9 Hz, 1H), 5.77 (d, J=10.1 Hz, 1H), 4.12 (s, 1H), 3.99 (s,
2H), 3.43-3.31 (m, 2H), 3.14 (d, J=28.5 Hz, 2H), 1.95 (s, 1H), 1.76
(s, 1H), 1.55 (s, 2H).
##STR00274## ##STR00275##
5-(pyridin-3-ylmethylthio)thiazol-2-amine (MFH-3-171-1)
[0521] To a solution of compound MFH-2-76-1 (300 mg, 1.91 mmol) in
EtOH (5 ml) was added NaBH.sub.4 (144 mg, 3.82 mmol) portionwise at
room temperature. The mixture was stirred for 1h, and then acetone
(3 ml) was slowly introduced. After 1 h, a solution of
3-(bromomethyl)pyridine hydrobromide (483 mg, 1.91 mmol) in EtOH (3
mL) was added. The resulting dark reaction mixture was heated to
reflux for 1 h, and was then cooled and concentrated in vacuo. The
residue was partitioned between EtOAc and brine. The organic phase
was separated, dried (MgSO.sub.4), and concentrated in vacuo to
give a crude solid which was triturated with diethyl ether/hexane
to provide compound MFH-3-171-1 (310 mg, 73%) LCMS (m/z): 224
[M+H]+.
2-bromo-5-(pyridin-3-ylmethylthio)thiazole (MFH-3-180-1)
[0522] To a solution of CuBr.sub.2 (370 mg, 1.66 mmol) in
acetonitrile (20 mL) at 0.degree. C. was added t-BuONO (172 mg,
1.66 mmol) followed by compound MFH-3-171-1 (310 mg, 1.39 mmol).
The mixture was stirred at 0.degree. C. for one hour, then at room
temperature for one hour, ethyl acetate was added and the organic
mixture washed with hydrochloric acid (2.times.50 mL), dried over
magnesium sulfate, filtered through a pad of silica gel, and
concentrated in vacuo. The residue was chromatographed on silica
gel to give the MFH-3-180-1 (288 mg, 72%). LCMS (m/z): 288
[M+H]+.
(R)-tert-butyl3-(5-(pyridin-3-ylmethylthio)thiazol-2-ylamino)piperidine-1--
carboxylate (MFH-3-182-1)
[0523] The mixture of MFH-3-180-1 (288 mg, 1 mmol), (R)-tert-butyl
3-aminopiperidine-1-carboxylate (320 mg, 1.6 mmol) and DIEA (194
mg, 1.5 mmol) in NMP (1 mL) was stirred at 140.degree. C. for
overnight. The residue was extracted with chloroform and
iso-propanol (4:1). The organic phase was washed with brine (20 mL)
and dried over Na.sub.2SO.sub.4. After removal of the solvent, the
residue was purified by silica gel (MeOH/DCM=0-20%) to obtain
MFH-3-182-1 (180 mg, yield 44%). LCMS (m/z): 407 [M+H].sup.+.
(R)--N-(piperidin-3-yl-5-(pyridin-3-ylmethylthio)thiazol-2-amine
(MFH-3-185-1)
[0524] To a solution of MFH-3-182-1 (180 mg, 0.44 mmol) in methanol
(3 mL) was added 4N HCl/dioxane (3 mL). The solution was then
stirred for 3h at room temperature and the solvent was removed
under reduced pressure to provide a crude which was directly used
in the next step. LCMS (m/z): 307 [M+H].sup.+.
(R)-(4-nitrophenyl)(3-(5-(pyridin-3-ylmethylthio)thiazol-2-ylamino)piperid-
in-1-yl)methanone (MFH-3-186-1)
[0525] The mixture of MFH-3-185-1 (120 mg, 0.39 mmol),
4-nitrobenzoyl chloride (72 mg, 0.39 mmol) in pyridine (2 mL) was
stirred for overnight at room temperature. Then the reaction
mixture was concentrated under reduced pressure and the residue was
directly used in the next step. LCMS (m/z): 456 [M+H].sup.+.
(R)-(4-aminophenyl)(3-(5-(pyridin-3-ylmethylthio)thiazol-2-ylamino)piperid-
in-1-yl)methanone (MFH-3-187-1)
[0526] To a solution of MFH-3-186-1 (178 mg, 0.39 mmol) in ethyl
acetate and methanol (1:1) were added Tin(II) chloride dehydrate
(881 mg, 3.9 mmol) and cone. HCl (0.2 mL). After stirring for 3 h
at 80.degree. C., the reaction mixture was diluted with chloroform
and iso-propanol (4:1), neutralized with saturated NaHCO.sub.3 and
filtered. The filtrate was extracted with chloroform and
iso-propanol (4:1), concentrated under reduced pressure and the
resulting residue was purified by silica gel column chromatography
(MeOH/DCM=0-20%) to give MFH-3-187-1 (20 mg, yield 12%). LCMS
(m/z): 426 [M+H]+.
(R)--N-(4-(3-(5-(pyridin-3-ylmethylthio)thiazol-2-ylamino)piperidine-1-car-
bonyl)phenyl)acrylamide (MFH-3-191-1)
[0527] To a solution of MFH-3-187-1 (20 mg, 0.05 mmol) and DIPEA
(0.1 mL) in CH.sub.3CN (2 mL) was added acryloyl chloride (6 mg,
0.06 mmol) in DCM (0.5 mL) dropwise. The mixture was then stirred
at 0.degree. C. for 1h. The solution was then concentrated under
reduced pressure and the residue was purified by prep-HPLC
(MeOH/H.sub.2O, 0.05% TFA) to provide MFH-3-191-1 (6 mg, yield
26%). LCMS (m/z): 480 [M+H]+. .sup.1H NMR (500 MHz, DMSO) .delta.
10.30 (s, 1H), 8.58 (d, J=53.1 Hz, 2H), 8.00 (d, J=44.0 Hz, 2H),
7.78-7.62 (m, 2H), 7.40 (d, J=29.7 Hz, 2H), 7.14 (dd, J=67.0, 35.1
Hz, 1H), 6.82 (s, 1H), 6.43 (dd, J=17.4, 10.0 Hz, 1H), 6.28 (t,
J=12.8 Hz, 1H), 5.84-5.74 (m, 1H), 4.04 (s, 1H), 3.98 (s, 2H), 3.31
(s, 2H), 3.17 (s, 2H), 1.96 (s, 1H), 1.75 (s, 1H), 1.54 (s,
2H).
##STR00276## ##STR00277##
(1R,3R)-3-(4-nitro-2-(trifluoromethyl)phenoxy)cyclohexanamine
(MFH-3-166-1)
[0528] To a solution of compound (1R,3R)-3-aminocyclohexanol
hydrochloride (120 mg, 0.8 mmol) in absolute DMF (1 ml) was added
NaH (130 mg, 3.3 mmol) portionwise at 0.degree. C. The mixture was
stirred for 1 h, and then
1-fluoro-4-nitro-2-(trifluoromethyl)benzene (170 mg, 0.8 mmol) was
slowly introduced. After 3h, the water was added slowly. The
residue was extracted with chloroform and iso-propanol (4:1). The
organic phase was washed with brine (20 mL.times.2) and dried over
Na.sub.2SO.sub.4. After removal of the solvent, the residue was
purified by silica gel (MeOH/DCM=0-20%) to obtain MFH-3-166-1 (50
mg, yield 20%). LCMS (m/z): 305 [M+H].sup.+.
5-((5-tert-butyloxazol-2-yl)methylthio)-N-((1R,3R)-3-(4-nitro-2-(trifluoro-
methyl)phenoxy)cyclohexyl)thiazol-2-amine (MFH-3-183-1)
[0529] The mixture of MFH-2-83-1 (55 mg, 0.16 mmol), MFH-3-166-1
(50 mg, 0.16 mmol) and DIEA (32 mg, 0.25 mmol) in NMP (0.5 mL) was
stirred at 140.degree. C. for overnight. The residue was extracted
with chloroform and iso-propanol (4:1). The organic phase was
washed with brine (20 mL.times.2) and dried over Na.sub.2SO.sub.4.
After removal of the solvent, the residue was purified by silica
gel (MeOH/DCM=0-20%) to obtain MFH-3-183-1 (28 mg, yield 31%). LCMS
(m/z): 557 [M+H].sup.+.
N-((1R,3R)-3-(4-amino-2-(trifluoromethyl)phenoxy)cyclohexyl)-5-((5-(tert-b-
utyloxazol-2-yl)methylthio)thiazol-2-amine (MFH-3-188-1)
[0530] To a solution of MFH-3-183-1 (28 mg, 0.05 mmol) in ethyl
acetate and methanol (1:1) were added Tin(II) chloride dehydrate
(113 mg, 0.5 mmol) and conc. HCl (0.1 mL). After stirring for 3 h
at 80.degree. C., the reaction mixture was diluted with chloroform
and iso-propanol (4:1), neutralized with saturated NaHCO.sub.3 and
filtered. The filtrate was extracted with chloroform and
iso-propanol (4:1), concentrated under reduced pressure and the
resulting residue was purified by silica gel column chromatography
(MeOH/DCM=0-20%) to give MFH-3-188-1 (20 mg, yield 75%). LCMS
(m/z): 527 [M+H]+.
N-(4-((1R,3R)-3-(5-(((5-tert-butyloxazol-2-yl)methylthio)thiazol-2-ylamino-
)cyclohexyloxy)-3-(trifluoromethyl)phenyl)acrylamide
(MFH-3-192-1)
[0531] To a solution of MFH-3-188-1 (20 mg, 0.04 mmol) and DIPEA
(0.1 mL) in CH.sub.3CN (2 mL) was added acryloyl chloride (5 mg,
0.05 mmol) in DCM (0.1 mL) dropwise. The mixture was then stirred
at 0.degree. C. for 1 h. The solution was then concentrated under
reduced pressure and the residue was purified by prep-HPLC
(MeOH/H.sub.2O, 0.05% TFA) to provide MFH-3-192-1 (5.4 mg, yield
24%). LCMS (m/z): 581 [M+H]+. .sup.1H NMR (500 MHz, DMSO) .delta.
10.27 (s, 1H), 8.04 (t, J=5.4 Hz, 2H), 7.82 (dd, J=9.0, 2.5 Hz,
1H), 7.25 (t, J=8.1 Hz, 1H), 6.95 (d, J=5.6 Hz, 1H), 6.70 (d, J=7.3
Hz, 1H), 6.39 (dd, J=17.0, 10.1 Hz, 1H), 6.26 (dd, J=17.0, 2.0 Hz,
1H), 5.77 (dd, J=10.1, 2.0 Hz, 1H), 4.92 (s, 1H), 3.93 (d, J=0.8
Hz, 2H), 2.10 (d, J=14.5 Hz, 1H), 1.93 (d, J=14.3 Hz, 1H), 1.79 (s,
1H), 1.65 (dd, J=22.7, 11.5 Hz, 2H), 1.59 (s, 2H), 1.32 (m, 1H),
1.23 (dd, J=11.2, 6.2 Hz, 1H), 1.17 (s, 9H).
##STR00278## ##STR00279##
5-((2-(trifluoromethyl)pyrimidin-5-yl)methylthio)thiazol-2-amine
(MFH-3-184-1)
[0532] To a solution of compound MFH-2-76-1 (300 mg, 1.91 mmol) in
absolute EtOH (5 ml) was added NaBH.sub.4 (144 mg, 3.82 mol) at
room temperature. The mixture was stirred for 1 h, and then acetone
(3 ml) was slowly introduced. After 1 h, a solution of
5-(chloromethyl)-2-(trifluoromethyl)pyrimidine (375 mg, 1.91 mmol)
in EtOH (3 ml) was added. The resulting dark reaction mixture was
heated to reflux for 1 h, and was then cooled and concentrated in
vacuo. The residue was partitioned between EtOAc and brine. The
organic phase was separated, dried (MgSO.sub.4), and concentrated
in vacuo to give a crude solid which was triturated with diethyl
ether/hexane to provide compound MFH-3-184-1 (500 mg, 90%) LCMS
(m/z): 293 [M+H]+.
2-bromo-5-((2-(trifluoromethyl)pyrimidin-5-yl)methylthio)thiazole
(MFH-3-189-1)
[0533] To a solution of CuBr.sub.2 (458 mg, 2.05 mmol) in
acetonitrile (15 mL) at 0.degree. C. was added t-BuONO (211 mg,
2.05 mmol) followed by compound MFH-3-184-1 (500 mg, 1.71 mmol).
The mixture was stirred at 0.degree. C. for one hour, then at room
temperature for one hour. Ethyl acetate was added and the organic
mixture was washed with hydrochloric acid (50 mL), dried over
magnesium sulfate, filtered through a pad of silica gel, and
concentrated in vacuo. The residue was chromatographed on silica
gel to give the MFH-3-189-1 (430 mg, 70%). LCMS (m/z): 357
[M+H]+.
(R)-tert-butyl3-(5-((2-(trifluoromethyl)pyrimidin-5-yl)methylthio)thiazol--
2-ylamino)piperidine-1-carboxylate (MFH-3-190-1)
[0534] The mixture of MFH-3-189-1 (430 mg, 1.21 mmol),
(R)-tert-butyl 3-aminopiperidine-1-carboxylate (362 mg, 1.81 mmol)
and DIEA (312 mg, 2.4 mmol) in NMP (1 mL) was stirred at
140.degree. C. for overnight. The residue was extracted with
chloroform and iso-propanol (4:1). The organic phase was washed
with brine (20 mL) and dried over Na.sub.2SO.sub.4. After removal
of the solvent, the residue was purified by silica gel
(MeOH/DCM=0-20%) to obtain MFH-3-190-1 (380 mg, yield 66%). LCMS
(m/z): 476 [M+H].sup.+.
(R)--N-(piperidin-3-yl)-5-((2-(trifluoromethyl)pyrimidin-5-yl)methylthio)t-
hiazol-2-amine (MFH-3-194-1)
[0535] To a solution of MFH-3-190-1 (380 mg, 0.8 mmol) in methanol
(5 mL) was added 4N HCl/dioxane (5 mL). The solution was then
stirred for 3h at room temperature and the solvent was removed
under reduced pressure to provide a crude which was directly used
in the next step. LCMS (m/z): 376 [M+H].sup.+.
(R)-(4-nitrophenyl)(3-(5-((2-(trifluoromethyl)pyrimidin-5-yl)methylthio)tr-
iazol-2-ylamino)piperidin-1-yl)methanone (MFH-3-195-1)
[0536] The mixture of MFH-3-194-1 (230 mg, 0.61 mmol),
4-nitrobenzoyl chloride (113 mg, 0.61 mmol) in pyridine (2 mL) was
stirred for overnight at room temperature. Then the reaction
mixture was concentrated under reduced pressure and the residue was
directly used in the next step. LCMS (m/z): 525 [M+H].sup.+.
(R)-(4-aminophenyl)(3-(5-((2-(trifluoromethyl)pyrimidin-5-yl)methylthio)th-
iazol-2-ylamino)piperidin-1-yl)methanone (MFH-3-198-1)
[0537] To a solution of MFH-3-195-1 (280 mg, 0.53 mmol) in ethyl
acetate and methanol (1:1) were added Tin(II) chloride dehydrate
(965 mg, 4.27 mmol) and cone. HCl (0.2 mL). After stirring for 3 h
at 80.degree. C., the reaction mixture was diluted with chloroform
and iso-propanol (4:1), neutralized with saturated NaHCO.sub.3 and
filtered. The filtrate was extracted with chloroform and
iso-propanol (4:1), concentrated under reduced pressure and the
resulting residue was purified by silica gel column chromatography
(MeOH/DCM=0-20%) to give MFH-3-198-1 (200 mg, yield 76%). LCMS
(m/z): 495 [M+H]+.
(R)--N-(4-(3-(5-((2-(trifluoromethyl)pyrimidin-5-yl)methylthio)thiazol-2-y-
lamino)piperidine-1-carbonyl)phenyl)acrylamide (MFH-3-201-1)
[0538] To a solution of MFH-3-198-1 (40 mg, 0.08 mmol) and DIPEA
(0.1 mL) in CH.sub.3CN (2 mL) was added acryloyl chloride (10 mg,
0.1 mmol) in DCM (0.2 mL) dropwise. The mixture was then stirred at
0.degree. C. for 1 h. The solution was then concentrated under
reduced pressure and the residue was purified by prep-HPLC
(MeOH/H.sub.2O, 0.05% TFA) to provide MFH-3-201-1 (9.8 mg, yield
22%). LCMS (m/z): 549 [M+H]+. .sup.1H NMR (500 MHz, DMSO) .delta.
10.26 (s, 1H), 8.81 (s, 2H), 8.11 (s, 1H), 7.79-7.59 (m, 2H), 7.38
(t, J=13.1 Hz, 2H), 6.99-6.73 (m, 1H), 6.50-6.35 (m, 1H), 6.34-6.17
(m, 1H), 5.80 (m, 1H), 4.01 (s, 2H), 3.81-3.73 (m, 3H), 3.45-3.20
(m, 2H), 1.94 (d, J=18.7 Hz, 1H), 1.78 (m, 1H), 1.54 (s, 2H).
##STR00280## ##STR00281##
5-(pyridin-2-ylmethylthio)thiazol-2-amine (MFH-3-170-1)
[0539] To a solution of compound MFH-2-7-1 (300 mg, 1.91 mmol) in
absolute EtOH (5 ml) was added NaBH.sub.4 (144 mg, 3.82 mmol)
portionwise at room temperature. The mixture was stirred for 1h,
and then acetone (3 ml) was slowly introduced. After 1 h, a
solution of 2-(chloromethyl)pyridine hydrochloride (313 mg, 1.91
mmol) in EtOH (3 ml) was added. The resulting dark reaction mixture
was heated to reflux for 1 h, and was then cooled and concentrated
in vacuo. The residue was partitioned between EtOAc and brine. The
organic phase was separated, dried (MgSO.sub.4), and concentrated
in vacuo to give a crude solid which was triturated with diethyl
ether/hexane to provide compound MFH-3-170-1 (260 mg, 61%) LCMS
(m/z): 224 [M+H]+.
2-bromo-5-(pyridin-2-ylmethylthio)thiazole (MFH-3-176-1)
[0540] To a solution of CuBr.sub.2 (313 mg, 1.4 mmol) in
acetonitrile (20 mL) at 0.degree. C. was added t-BuONO (144 mg, 1.4
mmol) followed by compound MFH-3-170-1 (260 mg, 1.16 mmol). The
mixture was stirred at 0.degree. C. for one hour, then at room
temperature for one hour. Ethyl acetate was added and the organic
mixture washed with hydrochloric acid (2.times.50 mL), dried over
magnesium sulfate, filtered through a pad of silica gel, and
concentrated in vacuo. The residue was chromatographed on silica
gel to give the MFH-3-176-1 (90 mg, 27%). LCMS (m/z): 288
[M+H]+.
(R)-tert-butyl3-(5-(pyridin-2-ylmethylthio)thiazol-2-ylamino)piperidine-1--
carboxylate (MFH-3-196-1)
[0541] The mixture of MFH-3-176-1 (180 mg, 0.63 mmol),
(R)-tert-butyl 3-aminopiperidine-1-carboxylate (200 mg, 1 mmol) and
DIEA (162 mg, 1.3 mmol) in NMP (I mL) was stirred at 140.degree. C.
for overnight. The residue was extracted with chloroform and
iso-propanol (4:1). The organic phase was washed with brine (20 mL)
and dried over Na.sub.2SO.sub.4. After removal of the solvent, the
residue was purified by silica gel (MeOH/DCM=0-20%) to obtain
MFH-3-196-1 (120 mg, yield 47%). LCMS (m/z): 407 [M+H].sup.+.
(R)--N-(piperidin-3-yl)-5-(pyridin-2-ylmethylthio)thiazol-2-amine
(MFH-3-197-1)
[0542] To a solution of MFH-3-196-1 (120 mg, 0.3 mmol) in methanol
(3 mL) was added 4N HCl/dioxane (3 mL). The solution was then
stirred for 3h at room temperature and the solvent was removed
under reduced pressure to provide a crude which was directly used
in the next step. LCMS (m/z): 307 [M+H].sup.+.
(R)-(4-nitrophenyl)(3-(5-(pyridin-2-ylmethylthio)thiazol-2-ylamino)piperid-
in-1-yl)methanone (MFH-3-199-1)
[0543] The mixture of MFH-3-197-1 (80 mg, 0.26 mmol),
4-nitrobenzoyl chloride (48 mg, 0.26 mmol) in pyridine (2 mL) was
stirred for overnight at room temperature. Then the reaction
mixture was concentrated under reduced pressure and the residue was
directly used in the next step. LCMS (m/z): 456 [M+H].sup.+.
(R)-(4-aminophenyl)(3-(5-(pyridin-2-ylmethylthio)thiazol-2-ylamino)piperid-
in-1-yl)methanone (MFH-3-200-1)
[0544] To a solution of MFH-3-199-1 (118 mg, 0.261 mmol) in ethyl
acetate and methanol (1:1) were added Tin(II) chloride dehydrate
(472 mg, 2.1 mmol) and conc. HCl (0.2 mL). After stirring for 3 h
at 80.degree. C., the reaction mixture was diluted with chloroform
and iso-propanol (4:1), neutralized with saturated NaHCO.sub.3 and
filtered. The filtrate was extracted with chloroform and
iso-propanol (4:1), concentrated under reduced pressure and the
resulting residue was purified by silica gel column chromatography
(MeOH/DCM=0-20%) to give MFH-3-200-1 (60 mg, yield 54%). LCMS
(m/z): 426 [M+H]+.
(R)--N-(4-(3-(5-(pyridin-3-ylmethylthio)thiazol-2-ylamino)piperidine-1-car-
bonyl)phenyl)acrylamide (MFH-3-191-1)
[0545] To a solution of MFH-3-187-1 (40 mg, 0.1 mmol) and DIPEA
(0.1 mL) in CH.sub.3CN (2 mL) was added acryloyl chloride (12 mg,
0.12 mmol) in DCM (0.2 mL) dropwise. The mixture was then stirred
at 0.degree. C. for 1h. The solution was then concentrated under
reduced pressure and the residue was purified by prep-HPLC
(MeOH/H.sub.2O, 0.05% TFA) to provide MFH-3-203-1 (8.4 mg, yield
19%). LCMS (m/z): 480 [M+H]+. .sup.1H NMR (500 MHz, DMSO) .delta.
10.30 (s, 1H), 8.62 (s, 1H), 8.06 (d, J=50.5 Hz, 2H), 7.72 (d,
J=21.6 Hz, 2H), 7.51 (s, 2H), 7.37 (s, 2H), 6.86 (s, 1H), 6.44 (dd,
J=17.1, 10.2 Hz, 1H), 6.27 (d, J=16.9 Hz, 1H), 5.85-5.74 (m, 1H).
4.04 (s, 1H), 3.98 (s, 2H), 3.31 (s, 2H), 3.17 (s, 2H), 1.96 (s,
1H), 1.75 (s, 1H), 1.54 (s, 2H).
##STR00282## ##STR00283##
5-((tetrahydro-2H-pyran-4-yl)methylthio)thiazol-2-amine
(MFH-3-202-1)
[0546] To a solution of compound MFH-2-76-1 (194 mg, 1.23 mmol) in
absolute EtOH (4 ml) was added NaBH.sub.4 (93 mg, 2.47 mmol) at
room temperature. The mixture was stirred for 1h, and then acetone
(2 ml) was slowly introduced. After 1h, a solution of
4-(bromomethyl)-tetrahydro-2H-pyran (221 mg, 1.23 mmol) in EtOH (2
ml) was added. The resulting dark reaction mixture was heated to
reflux for 1 h, and was then cooled and concentrated in vacuo. The
residue was partitioned between EtOAc and brine. The organic phase
was separated, dried (MgSO.sub.4), and concentrated in vacuo to
give a crude solid which was triturated with diethyl ether/hexane
to provide compound MFH-3-202-1 (250 mg, 88%) LCMS (m/z): 231
[M+H]+.
2-bromo-5-((tetrahydro-2H-pyran-4-yl)methylthio)thiazole
(MFH-3-204-1)
[0547] To a solution of CuBr.sub.2 (290 mg, 1.3 mmol) in
acetonitrile (15 mL) at 0.degree. C. was added t-BuONO (175 mg, 1.3
mmol) followed by compound MFH-3-202-1 (250 mg, 1.1 mmol). The
mixture was stirred at 0.degree. C. for one hour, then at room
temperature for one hour, ethyl acetate was added and the organic
mixture washed with hydrochloric acid (2.times.30 mL), dried over
magnesium sulfate, filtered through a pad of silica gel, and
concentrated in vacuo. The residue was chromatographed on silica
gel to give the MFH-3-204-1 (160 mg, 50%). LCMS (m/z): 295
[M+H]+.
(R)-tert-butyl3-(5-((tetrahydro-2H-pyran-4-yl)methylthio)thiazol-2-ylamino-
)piperidine-1-carboxylate (MFH-3-205-1)
[0548] The mixture of MFH-3-204-1 (160 mg, 0.54 mmol),
(R)-tert-butyl 3-aminopiperidine-1-carboxylate (174 mg, 0.87 mmol)
and DIEA (112 mg, 0.87 mmol) in NMP (0.5 mL) was stirred at
140.degree. C. for overnight. The residue was extracted with
chloroform and iso-propanol (4:1). The organic phase was washed
with brine (20 mL.times.2) and dried over Na.sub.2SO.sub.4. After
removal of the solvent, the residue was purified by silica gel
(MeOH/DCM=0-20%) to obtain MFH-3-205-1 (120 mg, yield 53%). LCMS
(m/z): 414 [M+H].sup.+.
(R)--N-(piperidin-3-yl)-5-((tetrahydro-2H-pyran-4-yl)methylthio)thiazol-2--
amine (MFH-3-206-1)
[0549] To a solution of MFH-3-205-1 (120 mg, 0.3 mmol) in methanol
(3 mL) was added 4N HCl/dioxane (3 mL). The solution was then
stirred for 3h at room temperature and the solvent was removed
under reduced pressure to provide a crude which was directly used
in the next step. LCMS (m/z): 314 [M+H].sup.+.
(R)-(4-nitrophenyl).sub.3-(5-((tetrahydro-2H-pyran-4-yl)methylthio)triazol-
-2-ylamino)piperidin-1-yl)methanone (MFH-3-207-1)
[0550] The mixture of MFH-3-206-1 (80 mg, 0.26 mmol),
4-nitrobenzoyl chloride (48 mg, 0.26 mmol) in pyridine (2 mL) was
stirred for overnight at room temperature. Then the reaction
mixture was concentrated under reduced pressure and the residue was
directly used in the next step. LCMS (m/z): 463 [M+H].sup.+.
(R)-(4-aminophenyl)(3-(5-((tetrahydro-2H-pyran-4-yl)methylthio)thiazol-2-y-
lamino)piperidin-1-yl)methanone (MFH-4-2-1)
[0551] To a solution of MFH-3-207-1 (120 mg, 0.26 mmol) in ethyl
acetate and methanol (1:1) were added Tin(II) chloride dehydrate
(472 mg, 2.1 mmol) and conc. HCl (0.2 mL). After stirring for 3 h
at 80.degree. C., the reaction mixture was diluted with chloroform
and iso-propanol (4:1), neutralized with saturated NaHCO.sub.3 and
filtered. The filtrate was extracted with chloroform and
iso-propanol (4:1), concentrated under reduced pressure and the
resulting residue was purified by silica gel column chromatography
(MeOH/DCM=0-20%) to give MFH-4-2-1 (60 mg, yield 54%). LCMS (m/z):
433 [M+H]+.
(R)--N-(4-(3-(5-((tetrahydro-2H-pyran-4-yl)methylthio)triazol-2-ylamino)pi-
peridine-1-carbonyl)phenyl)acrylamide (MFH-4-4-1)
[0552] To a solution of MFH-4-2-1 (30 mg, 0.07 mmol) and DIPEA (0.1
mL) in CH.sub.3CN (2 mL) was added acryloyl chloride (8 mg, 0.09
mmol) in DCM (0.1 mL) dropwise. The mixture was then stirred at
0.degree. C. for 1h. The solution was then concentrated under
reduced pressure and the residue was purified by prep-HPLC
(MeOH/H.sub.2O, 0.05% TFA) to provide MFH-4-4-1 (4.6 mg, yield
13%). LCMS (m/z): 487 [M+H]+. .sup.1H NMR (500 MHz, DMSO) .delta.
10.32 (d, J=28.3 Hz, 1H), 8.07 (s, 1H), 7.77-7.62 (m, 2H), 7.40
(dd, J=32.0, 7.9 Hz, 2H), 7.15-6.93 (m, 1H), 6.45 (dd, J=17.0, 10.1
Hz, 1H), 6.28 (dd, J=17.0, 1.9 Hz, 1H), 5.79 (dd, J=10.1, 1.9 Hz,
1H), 3.83 (d, J=9.4 Hz, 2H), 3.62 (s, 4H), 3.24 (t, J=11.1 Hz, 2H),
3.11 (s, 1H), 2.58 (d, J=6.4 Hz, 1H), 1.98 (d, J=12.6 Hz, 1H), 1.77
(s, 1H), 1.70 (d, J=11.6 Hz, 2H), 1.57 (m, 3H), 1.21 (m, 3H).
##STR00284## ##STR00285##
5-(pyrimidin-5-ylmethylthio)thiazol-2-amine (MFH-3-208-1)
[0553] To a solution of compound MFH-2-76-1 (230 mg, 1.46 mmol) in
absolute EtOH (5 ml) was added NaBH.sub.4 (166 mg, 4.38 mmol) at
room temperature. The mixture was stirred for 1h, and then acetone
(3 ml) was slowly introduced. After 1 h, a solution of
5-(chloromethyl)pyrimidine (241 mg, 1.46 mmol) in EtOH (3 ml) was
added. The resulting dark reaction mixture was heated to reflux for
1 h, and was then cooled and concentrated in vacuo. The residue was
partitioned between EtOAc and brine. The organic phase was
separated, dried (MgSO.sub.4), and concentrated in vacuo to give a
crude solid which was triturated with diethyl ether/hexane to
provide compound MFH-3-208-1 (180 mg, 55%) LCMS (m/z): 225
[M+H]+.
2-bromo-5-(pyrimidin-5-ylmethylthio)thiazole (MFH-4-1-1)
[0554] To a solution of CuBr.sub.2 (215 mg, 0.96 mmol) in
acetonitrile (15 mL) at 0.degree. C. was added t-BuONO (99 mg, 0.96
mmol) followed by compound MFH-3-208-1 (180 mg, 0.8 mmol). The
mixture was stirred at 0.degree. C. for one hour, then at room
temperature for one hour, ethyl acetate was added and the organic
mixture washed with hydrochloric acid (2.times.50 mL), dried over
magnesium sulfate, filtered through a pad of silica gel, and
concentrated in vacuo. The residue was chromatographed on silica
gel to give the MFH-4-1-1 (90 mg, 39%). LCMS (m/z): 289 [M+H]+.
(R)-tert-butyl3-(5-(pyrimidin-5-ylmethylthio)triazol-2-ylamino)piperidine--
1-carboxylate (MFH-4-3-1)
[0555] The mixture of MFH-4-1-1 (90 mg, 0.31 mmol), (R)-tert-butyl
3-aminopiperidine-1-carboxylate (100 mg, 0.5 mmol) and DIEA (65 mg,
0.5 mmol) in NMP (1 mL) was stirred at 140.degree. C. for
overnight. The residue was extracted with chloroform and
iso-propanol (4:1). The organic phase was washed with brine (20
mL.times.2) and dried over Na.sub.2SO.sub.4. After removal of the
solvent, the residue was purified by silica gel (MeOH/DCM=0-20%) to
obtain MFH-4-3-1 (66 mg, yield 52%). LCMS (m/z): 408
[M+H].sup.+.
(R)--N-(piperidin-3-yl)-5-(pyrimidin-5-ylmethylthio)thiazol-2-amine
(MFH-4-5-1)
[0556] To a solution of MFH-4-3-1 (66 mg, 0.16 mmol) in methanol (3
mL) was added 4N HCl/dioxane (3 mL). The solution was then stirred
for 3h at room temperature and the solvent was removed under
reduced pressure to provide a crude which was directly used in the
next step. LCMS (m/z): 308 [M+H].sup.+.
(R)-(4-nitrophenyl)(3-(5-(pyrimidin-5-ylmethylthio)thiazol-2-ylamino)piper-
idin-1-yl)methanone (MFH-4-6-1)
[0557] The mixture of MFH-4-5-1 (50 mg, 0.16 mmol), 4-nitrobenzoyl
chloride (30 mg, 0.16 mmol) in pyridine (2 mL) was stirred for
overnight at room temperature. Then the reaction mixture was
concentrated under reduced pressure and the residue was directly
used in the next step. LCMS (m/z): 457 [M+H].sup.+.
(R)-(4-aminophenyl)(3-(5-(pyrimidin-5-ylmethylthio)thiazol-2-ylamino)piper-
idin-1-yl)methanone (MFH-4-7-1)
[0558] To a solution of MFH-4-6-1 (74 mg, 0.16 mmol) in ethyl
acetate and methanol (1:1) were added Tin(II) chloride dehydrate
(292 mg, 1.3 mmol) and cone. HCl (0.1 mL). After stirring for 3 h
at 80.degree. C., the reaction mixture was diluted with chloroform
and iso-propanol (4:1), neutralized with saturated NaHCO.sub.3 and
filtered. The filtrate was extracted with chloroform and
iso-propanol (4:1), concentrated under reduced pressure and the
resulting residue was purified by silica gel column chromatography
(MeOH/DCM=0-20%) to give MFH-4-7-1 (15 mg, yield 22%). LCMS (m/z):
427 [M+H]+.
(R)--N-(4-(3-(5-(pyrimidin-5-ylmethylthio)thiazol-2-ylamino)piperidine-1-c-
arbonyl)phenyl)acrylamide (MFH-4-10-1)
[0559] To a solution of MFH-4-7-1 (15 mg, 0.04 mmol) and DIPEA (0.1
mL) in CH.sub.3CN (2 mL) was added acryloyl chloride (4 mg, 0.05
mmol) in DCM (0.1 mL) dropwise. The mixture was then stirred at
0.degree. C. for 1 h. The solution was then concentrated under
reduced pressure and the residue was purified by prep-HPLC
(MeOH/H.sub.2O, 0.05% TFA) to provide MFH-4-10-1 (2.2 mg, yield
13%). LCMS (m/z): 481 [M+H]+. .sup.1H NMR (500 MHz, DMSO) .delta.
10.26 (s, 1H), 9.07 (s, 1H) 8.81 (s, 2H), 8.11 (s, 1H), 7.79-7.59
(m, 2H), 7.38 (t, J=13.1 Hz, 2H), 6.99-6.73 (m, 1H), 6.50-6.35 (m,
1H), 6.34-6.17 (m, 1H), 5.80 (ddd, J=23.3, 10.0, 2.6 Hz, 1H), 4.01
(s, 2H), 3.81-3.73 (m, 3H), 3.45-3.20 (m, 2H), 1.94 (d, J=18.7 Hz,
1H), 1.78 (m, 1H), 1.54 (s, 2H).
##STR00286##
(R,E)-N-(4-(3-(5-((5-tert-butyloxazol-2-yl)methylthio)thiazol-2-ylamino)p-
iperidine-1-carbonyl)phenyl)-4-(dimethylamino)but-2-enamide
(MFH-4-13-1)
[0560] To a solution of (E)-4-bromobut-2-enoic acid (17 mg, 0.1
mmol) in SOCl.sub.2 (0.2 mL). The mixture was stirred at 70.degree.
C. for 1h under N.sub.2 atmosphere. The mixture was cooled to room
temperature and then was concentrated under reduced pressure. The
residue was diluted with dichloromethane and the resulted solution
was added dropwise to a solution of MFH-2-20-1 (37 mg, 0.08 mmol)
and DIPEA (0.2 mL) in CH.sub.3CN (2 mL) at 0.degree. C. After
stirring for 1 h at 0.degree. C., a solution of dimethylamine in
THF (2 mol/L, 0.1 ml, 0.2 mmol) was added and the reaction mixture
was stirred at room temperature for 1h. The removal of the solvent
under reduced pressure provided the residue which was purified by
HPLC (MeOH/H.sub.2O, 0.05% TFA) to obtain MFH-4-13-1 (18 mg, yield
39%). LCMS (m/z): 583 [M+H]+; .sup.1H NMR (500 MHz, DMSO) .delta.
10.18 (s, 1H), 7.96 (s, 1H), 7.66 (s, 2H), 7.34 (d, J=6.9 Hz, 2H),
6.84 (s, 1H), 6.80-6.65 (m, 2H), 6.27 (d, J=15.4 Hz, 1H), 3.94 (s,
2H), 3.79 (s, 1H), 3.65 (s, 2H), 3.14 (m, 2H), 3.06 (d, J=5.8, 2H),
2.18 (s, 6H), 1.95 (s, 1H), 1.75 (s, 1H), 1.51 (d, J=9.0 Hz, 2H),
1.17 (s, 9H).
##STR00287##
(R,E)-N-(4-(3-(5-((5-tert
butyloxazol-2-yl)methylthio)thiazol-2-ylamino)piperidin-1-ylsulfonyl)phen-
yl)-4-(dimethylamino)but-2-enamide (MFH-4-40-1)
[0561] To a solution of (E)-4-bromobut-2-enoic acid (12 mg, 0.07
mmol) in SOCl.sub.2 (0.2 mL). The mixture was stirred at 70.degree.
C. for 1h under N.sub.2 atmosphere. The mixture was cooled to room
temperature and then was concentrated under reduced pressure. The
residue was diluted with dichloromethane and the resulted solution
was added dropwise to a solution of MFH-4-40-1 (25 mg, 0.05 mmol)
and DIPEA (0.2 mL) in CH.sub.3CN (2 mL) at 0.degree. C. After
stirring for 1 h at 0.degree. C., a solution of dimethylamine in
THF (2 mol/L, 0.05 ml, 0.1 mmol) was added and the reaction mixture
was stirred at room temperature for 1h. The removal of the solvent
under reduced pressure provided the residue which was purified by
HPLC (MeOH/H.sub.2O, 0.05% TFA) to obtain MFH-4-40-1 (20 mg, yield
66%). LCMS (m/z): 619 [M+H].sup.+; .sup.1H NMR (500 MHz, DMSO)
.delta. 10.76 (s, 1H), 7.98 (d, J=7.2 Hz, 1H), 7.90 (dd, J=6.5, 4.7
Hz, 2H), 7.70 (dd, J=6.6, 4.8 Hz, 2H), 6.97 (d, J=2.7 Hz, 1H),
6.83-6.75 (m, 1H), 6.72 (d, J=8.8 Hz, 1H), 6.48 (d, J=15.3 Hz, 1H),
3.96 (s, 2H), 3.71 (s, 2H), 3.53 (d, J=9.9 Hz, 2H), 3.24 (d, J=11.2
Hz, 1H), 3.17 (s, 2H), 2.81 (s, 6H), 2.34-2.22 (m, 1H), 1.84-1.70
(m, 2H), 1.52 (dd, J=10.1, 4.0 Hz, 1H), 1.22 (s, 9H).
##STR00288## ##STR00289##
(R)-tert-butyl3-(5-((5-tert-butyloxazol-2-yl)methylthio)thiazol-2-ylamino-
)pyrrolidine-1-carboxylate (MFH-4-30-1)
[0562] The mixture of MFH-2-83-1 (150 mg, 0.45 mmol),
(R)-tert-butyl 3-aminopyrrolidine-1-carboxylate (134 mg, 0.72 mmol)
and DIEA (116 mg, 0.9 mmol) in NMP (1 mL) was stirred at
140.degree. C. for overnight. The residue was extracted with
chloroform and iso-propanol (4:1). The organic phase was washed
with brine (30 mL.times.2) and dried over Na.sub.2SO.sub.4. After
removal of the solvent, the residue was purified by silica gel
(MeOH/DCM=0-20%) to obtain MFH-4-30-1 (110 mg, yield 56%). LCMS
(m/z): 439 [M+H].sup.+.
(R)-5-((5-tert-butyloxazol-2-yl)methylthio)-N-(pyrrolidin-3-yl)thiazol-2-a-
mine (MFH-4-39-1)
[0563] To a solution of MFH-4-30-1 (110 mg, 0.25 mmol) in methanol
(3 mL) was added 4N HCl/dioxane (3 mL). The solution was then
stirred for 3h at room temperature and the solvent was removed
under reduced pressure to provide a crude which was directly used
in the next step. LCMS (m/z): 339 [M+H].sup.+.
(R)-(3-(5-((5-tert-butyloxazol-2-yl)methylthio)thiazol-2-ylamino)pyrrolidi-
n-1-yl)(4-nitrophenyl)methanone (MFH-4-41-1)
[0564] The mixture of MFH-4-39-1 (80 mg, 0.24 mmol), 4-nitrobenzoyl
chloride (53 mg, 0.28 mmol) in pyridine (2 mL) was stirred for
overnight at room temperature. Then the reaction mixture was
concentrated under reduced pressure and the residue was directly
used in the next step. LCMS (m/z): 488 [M+H].sup.+.
(R)-(4-aminophenyl)(3-(5-((5-tert-butyloxazol-2-yl)methylthio)triazol-2-yl-
amino)pyrrolidin-1-yl)methanone (MFH-4-44-1)
[0565] To a solution of MFH-4-41-1 (117 mg, 0.24 mmol) in ethyl
acetate and methanol (1:1) were added Tin(II) chloride dehydrate
(427 mg, 1.9 mmol) and conc. HCl (0.1 mL). After stirring for 3 h
at 80.degree. C., the reaction mixture was diluted with chloroform
and iso-propanol (4:1), neutralized with saturated NaHCO.sub.3 and
filtered. The filtrate was extracted with chloroform and
iso-propanol (4:1), concentrated under reduced pressure and the
resulting residue was purified by silica gel column chromatography
(MeOH/DCM=0-20%) to give MFH-4-44-1 (90 mg, yield 83%). LCMS (m/z):
458 [M+H]+.
(R)--N-(4-(3-(5-((5-tert-butyloxazol-2-yl)methylthio)thiazol-2-ylamino)pyr-
rolidine-1-carbonyl)phenyl)acrylamide (MFH-4-70-1)
[0566] To a solution of MFH-4-44-1 (22 mg, 0.05 mmol) and DIPEA
(0.2 mL) in CH.sub.3CN (2 mL) was added acryloyl chloride (5 mg,
0.06 mmol) in DCM (0.1 mL) dropwise. The mixture was then stirred
at 0.degree. C. for 1 h. The solution was then concentrated under
reduced pressure and the residue was purified by prep-HPLC
(MeOH/H.sub.2O, 0.05% TFA) to provide MFH-4-70-1 (13.4 mg, yield
54%). LCMS (m/z): 512 [M+H]+. .sup.1H NMR (500 MHz, DMSO) .delta.
10.33 (s, 1H), 8.38-8.21 (m, 1H), 7.73 (d, J=8.3 Hz, 2H), 7.52 (t,
J=8.6 Hz, 2H), 7.04-6.85 (m, 1H), 6.77-6.65 (m, 1H), 6.45 (dd,
J=17.0, 10.1 Hz, 1H), 6.29 (d, J=16.9 Hz, 1H), 5.84-5.75 (m, 1H),
3.96 (d, J=24.0 Hz, 2H), 3.82-3.72 (m, 1H), 3.57 (ddd, J=25.0,
12.1, 7.0 Hz, 3H), 3.33 (d, J=7.5 Hz, 1H), 2.15 (dt, J=30.0, 14.1
Hz, 1H), 1.92 (m, 1H), 1.23-1.12 (m, 9H).
##STR00290##
(R,E)-N-(4-(3-(5-((5-tert-butyloxazol-2-yl)methylthio)thiazol-2-ylamino)p-
yrrolidine-1-carbonyl)phenyl)-4-dimethylamino)but-2-enamide
(MFH-4-73-1)
[0567] To a solution of (E)-4-bromobut-2-enoic acid (15 mg, 0.09
mmol) in SOCl.sub.2 (0.2 mL). The mixture was stirred at 70.degree.
C. for 1h under N.sub.2 atmosphere. The mixture was cooled to room
temperature and then was concentrated under reduced pressure. The
residue was diluted with dichloromethane and the resulted solution
was added dropwise to a solution of MFH-4-40-1 (30 mg, 0.07 mmol)
and DIPEA (0.2 mL) in CH.sub.3CN (2 mL) at 0.degree. C. After
stirring for 1 h at 0.degree. C., a solution of dimethylamine in
THF (2 mol/L, 0.1 ml, 0.2 mmol) was added and the reaction mixture
was stirred at room temperature for 1h. The removal of the solvent
under reduced pressure provided the residue which was purified by
HPLC (MeOH/H.sub.2O, 0.05% TFA) to obtain MFH-4-73-1 (23 mg, yield
61%). LCMS (m/z): 569 [M+H].sup.+; .sup.1H NMR (500 MHz, DMSO)
.delta. 10.52 (s, 1H), 8.30-8.19 (m, 1H), 7.73 (d, J=8.3 Hz, 2H),
7.53 (t, J=8.4 Hz, 2H), 7.03-6.85 (m, 1H), 6.78 (dd, J=15.1, 7.4
Hz, 1H), 6.75-6.65 (m, 1H), 6.48 (d, J=15.3 Hz, 1H), 3.97 (s, 2H),
3.93 (s, 2H), 3.81-3.71 (m, 1H), 3.65-3.42 (m, 3H), 3.32 (dd,
J=10.0, 2.5 Hz, 1H), 2.81 (d, J=2.5 Hz, 6H), 2.21-2.10 (m, 1H),
1.99-1.85 (m, 1H), 1.23-1.12 (m, 9H).
##STR00291##
(4-aminophenyl)((3R)-3-((5-((5-(tert-butyl)oxazol-2-yl)methyl)sulfinyl)th-
iazol-2-yl)amino)piperidin-1-yl)methanone (YLIU-01-006-1)
[0568] To a solution of MFH-2-89-1 (30 mg, 0.064 mmol) in DCM (2
mL) was added m-CPBA (12 mg, 0.07 mmol), the reaction mixture was
stirred at rt overnight. Then diluted with DCM, washed with
NaHCO.sub.3 aq.(sat.), concentrated and purified with silica gel
column (eluted with MeOH in DCM 0% to 15%) to give the title
compound (20 mg, 64%) as a brown oil. LCMS (m/z): 488 [M+H]+.
N-(4-((3R)-3-((5-(((5-(tert-butyl)oxazol-2-yl)methyl)sulfinyl)thiazol-2-yl-
)amino)piperidine-1-carbonyl)phenyl)acrylamide (YLIU-01-007-1)
[0569] To a solution of YLIU-01-006-1 (20 mg, 0.04 mmol) in MeCN (2
mL) was added DIPEA (25 mg, 0.2 mmol). At 0.degree. C., acryloyl
chloride (4 mg, 0.044 mmol) was added dropwise. Monitored with
LCMS, when the reaction was completed, Na.sub.2CO.sub.3 aq.(sat.)
was added, the resulting mixture was extracted with DCM/i-PrOH
(4/1) (20 mL). The combined organic layer was concentrated and
purified with Prep-HPLC (MeOH/H.sub.2O, 0.05% TFA) to give the
title compound (8 mg, 36.9%) as white solid. LCMS (m/z): 542
[M+H]+.
[0570] 1H NMR (500 MHz, DMSO) .delta. 10.18 (s, 1H), 8.45 (s, 1H),
7.71-7.48 (m, 2H), 7.39-7.10 (m, 3H), 6.68 (s, 1H), 6.42-6.24 (m,
1H), 6.23-6.06 (m, 1H), 5.77-5.57 (m, 1H), 4.62-4.44 (m, 2H),
4.42-4.20 (m, 2H), 3.56 (m, 3H), 2.02-1.68 (m, 2H), 1.57-1.32 (m,
2H), 0.99 (s, 9H).
##STR00292## ##STR00293##
5-Thiocyanatothiazol-2-amine (YLIU-01-004-1)
[0571] A mixture of 2-amino-5-bromothiazole hydrobromide (7.8 g, 30
mmol) and potassium thiocyanate (20.46 g, 210.93 mmol) in methanol
(230 mL) was stirred at room temperature for 48 h. Methanol was
evaporated and water (30 ml) was added. The pH of the aqueous
solution was adjusted to pH=12 with 10% NaOH aq and precipitate
formed. The solid was collected by filtration to yield compound
YLIU-01-004-1 (3.3 g, 70%) as a brownish solid. LCMS (m/z): 158
[M+H]+.
5-((5-tert-Butyloxazol-2-yl)methylthio)thiazol-2-amine
(YLIU-01-005-1)
[0572] To a solution of compound YLIU-01-004-1 (156 mg, 1 mmol) in
absolute EtOH (3 ml) was added NaBH.sub.4 (76 mg, 2 mmol)
portionwise at 0.degree. C. The mixture was stirred at rt for 1 h,
and then acetone (2 ml) was slowly introduced. After 1 h, a
solution of 2-(bromomethyl)-5-(tert-butyl)oxazole (240 mg, 1.1
mmol) in EtOH (2 ml) was added. The resulting dark reaction mixture
was heated to 65.degree. C. for 1 h, and was then cooled and
concentrated in vacuo. The residue was partitioned between EtOAc
and brine. The organic phase was separated, dried (MgSO4),
concentrated and purified with silica gel column (eluted with MeOH
in DCM 0% to 10%) to provide the title compound (120 mg, 44%) as a
brown solid. LCMS (m/z): 270 [M+H]+.
2-(((2-bromothiazol-5-yl)thio)methyl)-5-(tert-butyl)oxazole
(YLIU-01-017-1)
[0573] To a solution of CuBr.sub.2 (2.32 g, 10 mmol) in
acetonitrile (50 mL) at 0.degree. C. was added t-BuONO (1.1 g, 10
mmol) followed by compound YLIU-01-004-1 (1.66 g, 6.13 mmol). The
mixture was stirred at 0.degree. C. for 1 h, then at room
temperature for 1 h. Ethyl acetate was added and the organic layer
was washed with hydrochloric acid (2.times.50 mL), dried over
magnesium sulfate, filtered through a pad of silica gel, and
concentrated in vacuo. The residue was purified on silica gel
column to give the YLIU-01-017-1 (690 mg, 33%) as orange oil. LCMS
(m/z): 333 [M+H]+.
tert-butyl
(R)-3-((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl-
)amino) piperidine-1-carboxylate (YLIU-01-038-1)
[0574] A mixture of YLIU-01-017-1 (150 mg, 0.45 mmol), tert-butyl
(R)-3-aminopiperidine-1-carboxylate (180 mg, 0.9 mmol), DIPEA (0.24
mL, 1.35 mmol) in NMP (3 mL) was heated to 140.degree. C.
overnight. The solution was diluted with water (20 mL) and
extracted with chloroform and iso-propanol (4:1). The organic phase
was washed with brine (50 mL.times.2) and dried over
Na.sub.2SO.sub.4, filtered and concentrated. The residue was
purified by silica gel column (MeOH/DCM, 0-20%) to give
YLIU-01-038-1 (150 mg, 73%). LCMS (m/z): 453 [M+H]+.
(R)-5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)-N-(piperidin-3-yl)thiazol--
22-amine (YLIU-01-047-1)
[0575] To a mixture of compound YLIU-01-038-1 (150 mg, 0.33 mmol)
in methanol (2 mL) was added 4N HCl/dioxane (2 mL) and the resulted
solution was stirred at rt for 3 h. The mixture was concentrated
under reduced pressure to give the title compound as HCl salt,
which was directly used in the next step. LCMS (m/z): 353
[M+H].sup.+.
(R)-(3-((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)amino)pip-
eridin-1-yl)(5-nitropyridin-2-yl)methanone (YLIU-01-048-1)
[0576] To a mixture of YLIU-01-047-1 (50 mg, 0.12 mmol),
5-nitropicolinic acid (23 mg, 0.14 mmol) and DIPEA (78 mg, 0.6
mmol) in DCM (2 mL) was added T.sub.3P (50% solution in EA, 230 mg,
0.36 mmol) dropwise at rt. The reaction mixture was stirred at rt
overnight. The solution was diluted with DCM (20 mL) and washed
with brine (50 mL.times.2) and dried over Na.sub.2SO.sub.4,
filtered and concentrated. The residue was purified by silica gel
column (MeOH/DCM, 0-20%) to give the title compound (46 mg, 76%) as
yellow solid. LCMS (m/z): 503 [M+H].sup.+.
(R)-(5-aminopyridin-2-yl)(3-((5-((5-(tertbutyl)oxazol-2-yl)methyl)thio)thi-
azol-2-yl)amino)piperidin-1-yl)methanone (YLIU-01-064-1)
[0577] To a solution of YLIU-01-048-1 (46 mg, 0.091 mmol) in ethyl
acetate and methanol (2 mL, 1:1) was added Tin(II) chloride (138
mg, 0.73 mmol) at rt. After stirring for 2 h at 80.degree. C., the
reaction mixture was diluted with chloroform and iso-propanol
(4:1), neutralized with saturated NaHCO.sub.3 aq. and filtered. The
filtrate was extracted with chloroform and iso-propanol (4:1). The
combined organic layer was concentrated under reduced pressure and
the resulting residue was purified by silica gel column
chromatography (MeOH/DCM=0-20%) to give YLIU-01-064-1 (16 mg, 37%)
as yellow solid. LCMS (m/z): 473 [M+H]+.
(R)--N-(6-(3-((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)ami-
no)piperidine-1-carbonyl)pyridin-3-yl)acrylamide
(YLIU-01-067-1)
[0578] To a solution of YLIU-01-064-1 (16 mg, 0.034 mmol) and DIPEA
(13 mg, 0.1 mmol) in MeCN (2 mL) was added acryloyl chloride (4 mg,
0.044 mmol) dropwise at 0.degree. C. Monitored with LCMS, when the
reaction was completed, Na.sub.2CO.sub.3 aq.(sat.) was added, the
resulting mixture was extracted with DCM/i-PrOH (4/1) (20
mL.times.2). The combined organic layer was concentrated and
purified with Prep-HPLC (MeOH/H.sub.2O, 0.05% TFA) to give the
title compound (1.3 mg, 7.27%) as white solid. LCMS (m/z): 527
[M+H]+.
##STR00294## ##STR00295##
tert-butyl
4-((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)amino)piperid-
ine-1-carboxylate (YLIU-01-062-1)
[0579] A mixture of YLIU-01-017-1 (150 mg, 0.45 mmol), tert-butyl
4-aminopiperidine-1-carboxylate (180 mg, 0.9 mmol), DIPEA (0.24 mL,
1.35 mmol) in NMP (3 mL) was heated to 140.degree. C. overnight.
The solution was diluted with water (20 mL) and extracted with
chloroform and iso-propanol (4:1). The organic phase was washed
with brine (50 mL.times.2) and dried over Na.sub.2SO.sub.4,
filtered and concentrated. The residue was purified by silica gel
column (MeOH/DCM, 0-20%) to give YLIU-01-062-1 (150 mg, 73%). LCMS
(m/z): 453 [M+H]+.
5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)-N-(piperidin-4-yl)thiazol-2-am-
ine (YLIU-01-062A-1)
[0580] To a mixture of compound YLIU-01-062-1 (150 mg, 0.33 mmol)
in methanol (2 mL) was added 4N HCl/dioxane (2 mL) and the resulted
solution was stirred at rt for 3 h. The mixture was concentrated
under reduced pressure to give the title compound as HCl salt,
which was directly used in the next step. LCMS (m/z): 353
[M+H].sup.+.
(4-((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)amino)piperid-
in-1-yl)(3-nitrophenyl)methanone (YLIU-01-068-1)
[0581] The mixture of YLIU-01-062A-1 (330 mg, 0.89 mmol),
3-nitrobenzoyl chloride (330 mg, 1.78 mmol) in pyridine (2 mL) was
stirred at rt overnight. Then the reaction mixture was concentrated
under reduced pressure and the residue was purified by silica gel
column (MeOH/DCM, 0-20%) to give YLIU-01-068-1 (320 mg, 72%) as
yellow solid. LCMS (m/z): 502 [M+H]+.
(3-aminophenyl)(4-((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-y-
l)amino)piperidin-1-yl)methanone (YLIU-01-076-1)
[0582] To a solution of YLIU-01-068-1 (100 mg, 0.2 mmol) in MeOH (3
mL) was added Pd/C (10%, 20 mg) and N.sub.2H4-H.sub.2O (0.1 mL).
The reaction mixture was stirred at 80.degree. C. overnight, then
filtered through Celite. The filtrate was concentrated and purified
by Prep-HPLC (MeOH/H.sub.2O, 0.05% TFA) to give the title compound
(27 mg, 29%) as yellow oil. LCMS (m/z): 472 [M+H]+.
N-(3-(4-((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)amino)pi-
peridine-1-carbonyl)phenyl)acrylamide (YLIU-01-078-1)
[0583] To a solution of YLIU-01-076-1 (20 mg, 0.042 mmol) and DIPEA
(17 mg, 0.13 mmol) in MeCN (2 mL) was added acryloyl chloride (4.2
mg, 0.046 mmol) dropwise at 0.degree. C. Monitored with LCMS, when
the reaction was completed, Na.sub.2CO.sub.3 aq.(sat.) was added,
the resulting mixture was extracted with DCM/i-PrOH (4/1) (20
mL.times.2). The combined organic layer was concentrated and
purified with Prep-HPLC (MeOH/H.sub.2O, 0.05% TFA) to give the
title compound (1.4 mg, 6.3%) as white solid. LCMS (m/z): 526
[M+H]+.
##STR00296## ##STR00297##
4-methyl-5-thiocyanatothiazol-2-amine (YLIU-01-056-1)
[0584] A mixture of 2-amino-5-bromothiazole hydrobromide (580 mg, 3
mmol) and potassium thiocyanate (2.9 g, 30 mmol) in methanol (20
mL) was stirred at room temperature for 48 h. Methanol was
evaporated and water (3 ml) was added. The pH of the aqueous
solution was adjusted to pH=12 with 10% NaOH aq and precipitate
formed. The solid was collected by filtration to yield compound
YLIU-01-056-1 (480 mg, 93%) as a brownish solid. LCMS (m/z): 172
[M+H]+.
5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)-4-methylthiazol-2-amine
(YLIU-01-073-1)
[0585] To a solution of compound YLIU-01-056-1 (171 mg, 1 mmol) in
absolute EtOH (3 ml) was added NaBH.sub.4 (76 mg, 2 mmol)
portionwise at 0.degree. C. The mixture was stirred at rt for 1 h,
and then acetone (2 ml) was slowly introduced. After 1 h, a
solution of 2-(bromomethyl)-5-(tert-butyl)oxazole (240 mg, 1.1
mmol) in EtOH (2 ml) was added. The resulting dark reaction mixture
was heated to 65.degree. C. for 1 h, and was then cooled and
concentrated in vacuo. The residue was partitioned between EtOAc
and brine. The organic phase was separated, dried (MgSO4),
concentrated and purified with silica gel column (eluted with MeOH
in DCM 0% to 10%) to provide the title compound (314 mg, >100%)
as a brown oil. LCMS (m/z): 284 [M+H]+.
2-(((2-bromo-4-methylthiazol-5-yl)thio)methyl)-5-(tert-butyl)-4-methyloxaz-
ole (YLIU-01-089-1)
[0586] To a solution of CuBr.sub.2 (423 mg, 1.9 mmol) in
acetonitrile (10 mL) at 0.degree. C. was added t-BuONO (200 mg, 1.9
mmol) followed by compound YLIU-01-073-1 (314 mg, 1.1 mmol). The
mixture was stirred at 0.degree. C. for 1 h, then at room
temperature for 1 h. Ethyl acetate was added and the organic layer
was washed with hydrochloric acid (2.times.50 mL), dried over
magnesium sulfate, filtered through a pad of silica gel, and
concentrated in vacuo. The residue was purified on silica gel
column to give the YLIU-01-089-1 (140 mg, 37%) as orange oil. LCMS
(m/z): 347 [M+H]+.
tert-butyl
(R)-3-((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)-4-methylthi-
azol-2-yl) amino)piperidine-1-carboxylate (YLIU-01-092-1)
[0587] A mixture of YLIU-01-089-1 (140 mg, 0.4 mmol), tert-butyl
(R)-3-aminopiperidine-1-carboxylate (160 mg, 0.8 mmol), DIPEA (0.22
mL, 1.2 mmol) in NMP (3 mL) was heated to 140.degree. C. overnight.
The solution was diluted with water (20 mL) and extracted with
chloroform and iso-propanol (4:1). The organic phase was washed
with brine (50 mL.times.2) and dried over Na.sub.2SO.sub.4,
filtered and concentrated. The residue was purified by silica gel
column (MeOH/DCM, 0-20%) to give YLIU-01-092-1 (220 mg, >100%).
LCMS (m/z): 467 [M+H]+.
(R)-5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)-4-methyl-N-(piperidin-3-yl-
)thiazol-2-amine (YLIU-01-096-1)
[0588] To a mixture of compound YLIU-01-092-1 (220 mg, 0.47 mmol)
in methanol (2 mL) was added 4N HCl/dioxane (2 mL) and the resulted
solution was stirred at rt for 3 h. The mixture was concentrated
and the residue was purified by Prep-HPLC (MeOH/H.sub.2O, 0.05%
TFA) to give the title compound (88 mg, 51%) as yellow solid. LCMS
(m/z): 367 [M+H]+.
(R)--N-(4-(3-((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)-4-methylthiazol-
-2-yl)amino)piperidine-1-carbonyl)phenyl)acrylamide
(YLIU-01-099-1)
[0589] To a mixture of YLIU-01-096-1 (22 mg, 0.06 mmol),
4-acrylamidobenzoic acid (14 mg, 0.072 mmol) and DIPEA (39 mg, 0.3
mmol) in DCM (2 mL) was added T.sub.3P (50% solution in EA, 57 mg,
0.18 mmol) dropwise at rt. The reaction mixture was stirred at rt
overnight. The solution was diluted with DCM (20 mL) and washed
with brine (50 mL.times.2) and dried over Na.sub.2SO.sub.4,
filtered and concentrated. The residue was purified by Prep-HPLC
(MeOH/H.sub.2O, 0.05% TFA) to give the title compound (17.1 mg,
53%) as yellow solid. LCMS (m/z): 540 [M+H].sup.+.
[0590] 1H NMR (500 MHz, DMSO) .delta. 10.28 (s, 1H), 8.44-7.89 (m,
1H), 7.81-7.57 (m, 2H), 7.36 (m, 2H), 6.81-6.60 (m, 1H), 6.51-6.36
(m, 1H), 6.32-6.20 (m, 1H), 5.88-5.67 (m, 1H), 3.87 (m, 3H), 3.63
(s, 2H), 3.12 (m, 2H), 1.90 (m, 2H), 1.63-1.39 (m, 5H), 1.19 (s,
9H).
##STR00298##
tert-butyl
5-((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)amino)-3,3-di-
fluoropiperidine-1-carboxylate (YLIU-01-101-1)
[0591] A mixture of YLIU-01-017-1 (130 mg, 0.4 mmol), tert-butyl
5-amino-3,3-difluoropiperidine-1-carboxylate (350 mg, 1.5 mmol),
DIPEA (0.36 mL, 1.8 mmol) in NMP (3 mL) was heated to 140.degree.
C. overnight. The solution was diluted with water (20 mL) and
extracted with chloroform and iso-propanol (4:1). The organic phase
was washed with brine (50 mL.times.2) and dried over
Na.sub.2SO.sub.4, filtered and concentrated. The residue was
purified by silica gel column (MeOH/DCM, 0-20%) to give
YLIU-01-101-1 (12 mg, 6%). LCMS (m/z): 489 [M+H]+.
5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)-N-(5,5-difluoropiperidin-3-yl)-
triazol-2-amine (YLIU-01-111-1)
[0592] To a mixture of compound YLIU-01-101-1 (12 mg, 0.024 mmol)
in methanol (2 mL) was added 4N HCl/dioxane (2 mL) and the resulted
solution was stirred at rt for 3 h. The mixture was concentrated
and the crude product was used into next step directly as HCl salt.
LCMS (m/z): 389 [M+H]+.
N-(4-((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)amino)-3,3--
difluoropiperidine-1-carbonyl)phenyl)acrylamide (YLIU-01-114-1)
[0593] To a mixture of YLIU-01-111-1 (10 mg, crude HCl salt),
4-acrylamidobenzoic acid (6 mg, 0.03 mmol) and DIPEA (16 mg, 0.12
mmol) in DCM (2 mL) was added T.sub.3P (50% solution in EA, 46 mg,
0.07 mmol) dropwise at rt. The reaction mixture was stirred at rt
overnight. The solution was diluted with DCM (20 mL) and washed
with brine (50 mL.times.2) and dried over Na.sub.2SO.sub.4,
filtered and concentrated. The residue was purified by Prep-HPLC
(MeOH/H.sub.2O, 0.05% TFA) to give the title compound (2.5 mg, 19%)
as yellow solid. LCMS (m/z): 562 [M+H]+.
[0594] 1H NMR (500 MHz, DMSO) .delta. 10.35 (s, 1H), 8.10 (s, 1H),
7.74 (d, J=8.3 Hz, 2H), 7.42 (d, J=8.6 Hz, 2H), 7.08-6.83 (m, 1H),
6.72 (m, 1H), 6.45 (m, 1H), 6.29 (m, 1H), 5.80 (m, 1H), 3.97 (s,
2H), 3.65-3.44 (m, 3H), 3.22-2.90 (m, 2H), 2.08 (m, 2H), 1.21 (s,
9H).
##STR00299##
Methyl 4-(N-acryloylacrylamido)-3-(trifluoromethyl)benzoate
(YLIU-01-100-1)
[0595] To a solution of methyl 4-amino-3-(trifluoromethyl)benzoate
(220 mg, 1 mmol) and DIPEA (390 mg, 3 mmol) in MeCN (5 mL) was
added acryloyl chloride (108 mg, 1.2 mmol) dropwise at 0.degree. C.
The reaction was stirred at rt for 1 h, then Na.sub.2CO.sub.3
aq.(sat.) was added, the resulting mixture was extracted with DCM
(20 mL.times.2). The combined organic layer was concentrated and
purified with SGC (Hexane/EA-4/1) to give YLIU-01-100-1 (114 mg,
35%) as white solid. LCMS (m/z): 328 [M+H]+.
4-acrylamido-3-(trifluoromethyl)benzoic acid (YLIU-01-115-1)
[0596] To a solution of YLIU-01-100-1 (114 mg, 0.35 mmol) in THF (4
mL) was added 1N LiOH aq. (4 mL). The reaction mixture was stirred
at rt for 1h, then adjusted PH<7 with 4N HCl aq. The resulting
mixture was extracted with EA (20 mL.times.3), washed with brine
(50 mL.times.2) and dried over Na.sub.2SO.sub.4, filtered and
concentrated to give the desire product (110 mg, >100%) as a
white solid. LCMS (m/z): 260 [M+H]+.
(R)--N-(4-(3-((4-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)ami-
no)piperidine-1-carbonyl)-2-(trifluoromethyl)phenyl)acrylamide
(YLIU-01-121-1)
[0597] To a mixture of YLIU-01-047-1 (20 mg, 0.057 mmol),
YLIU-01-115-1 (18 mg, 0.068 mmol) and DIPEA (37 mg, 0.285 mmol) in
DCM (2 mL) was added T.sub.3P (50% solution in EA, 108 mg, 0.17
mmol) dropwise at rt. The reaction mixture was stirred at rt
overnight. The solution was diluted with DCM (20 mL) and washed
with brine (50 mL.times.2) and dried over Na.sub.2SO.sub.4,
filtered and concentrated. The residue was purified by Prep-HPLC
(MeOH/H.sub.2O, 0.05% TFA) to give the title compound (17.8 mg,
53%) as yellow solid. LCMS (m/z): 594 [M+H].sup.+. .sup.1H NMR (500
MHz, DMSO) .delta. 9.87 (m, 1H), 8.15 (m, 1H), 7.72 (m, 3H),
7.08-6.66 (m, 2H), 6.64-6.50 (m, 1H), 6.29 (m, 1H), 5.93-5.70 (m,
1H), 3.94 (m, 2H), 3.72 (m, 2H), 3.40 (m, 1H), 3.15 (m, 2H),
2.07-1.66 (m, 2H), 1.64-1.35 (m, 2H), 1.14 (s, 9H).
##STR00300##
methyl 4-acrylamido-3-fluorobenzoate (YLIU-01-116-1)
[0598] To a solution of methyl 4-amino-3-fluorobenzoate (500 mg, 3
mmol) and DIPEA (1.65 mL, 9 mmol) in MeCN (5 mL) was added acryloyl
chloride (320 mg, 3.6 mmol) dropwise at 0.degree. C. The reaction
was stirred at rt for 1 h, then Na.sub.2CO.sub.3 aq.(sat.) was
added, the resulting mixture was extracted with DCM (20
mL.times.2). The combined organic layer was concentrated and
recrystallized from EA to give YLIU-01-116-1 (190 mg, 28%) as white
solid. LCMS (m/z): 224 [M+H]+.
4-acrylamido-3-fluorobenzoic acid (YLIU-01-119-1)
[0599] To a solution of YLIU-01-116-1 (190 mg, 0.85 mmol) in
THF/water (4 mL/4 mL) was added LiOH (204 mg, 8.5 mmol). The
reaction mixture was stirred at rt for 1h, then adjusted PH<7
with 4N HCl aq. The resulting mixture was extracted with EA (20
mL.times.3), washed with brine (50 mL.times.2) and dried over
Na.sub.2SO.sub.4, filtered and concentrated to give the desire
product (110 mg, 61%) as a white solid. LCMS (m/z): 210 [M+H]+.
(R)--N-(4-(3-((4-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)ami-
no)piperidine-1-carbonyl)-2-fluorophenyl)acrylamide
(YLIU-01-123-1)
[0600] To a mixture of YLIU-01-047-1 (20 mg, 0.057 mmol),
YLIU-01-115-1 (15 mg, 0.068 mmol) and DIPEA (37 mg, 0.285 mmol) in
DCM (2 mL) was added T.sub.3P (50% solution in EA, 108 mg, 0.17
mmol) dropwise at rt. The reaction mixture was stirred at rt
overnight. The solution was diluted with DCM (20 mL) and washed
with brine (50 mL.times.2) and dried over Na.sub.2SO.sub.4,
filtered and concentrated. The residue was purified by Prep-HPLC
(MeOH/H.sub.2O, 0.05% TFA) to give the title compound (23.8 mg,
77%) as yellow solid. LCMS (m/z): 544 [M+H].sup.+. .sup.1H NMR (500
MHz, DMSO) .delta. 10.06 (s, 1H), 8.05 (m, 2H), 7.44-6.97 (m, 2H),
6.74-6.53 (m, 3H), 6.36-6.21 (m, 1H), 5.80 (m, 1H), 3.69 (s, 2H),
3.53 (m, 3H), 3.31-3.01 (m, 2H), 1.86 (m, 2H), 1.51 (m, 2H), 1.16
(s, 9H).
##STR00301##
benzyl
(2R,3R)-3-((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)triazol-2-y-
l)amino)-2-methylpiperidine-1-carboxylate (YLIU-01-142-1)
[0601] A mixture of YLIU-01-017-1 (80 mg, 0.24 mmol), benzyl
(2R,3R)-3-amino-2-methyl piperidine-1-carboxylate (300 mg, 1.2
mmol), DIPEA (0.42 mL, 2.4 mmol) in NMP (3 mL) was heated to
140.degree. C. overnight. The solution was diluted with water (20
mL) and extracted with chloroform and iso-propanol (4:1). The
organic phase was washed with brine (50 mL.times.2) and dried over
Na.sub.2SO.sub.4, filtered and concentrated. The residue was
purified by silica gel column (MeOH/DCM, 0-20%) to give
YLIU-01-142-1 (70 mg, 58%) as yellow solid. LCMS (m/z): 501
[M+H]+.
5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)-N-((2R,3R)-2-methylpiperidin-3-
-yl)triazol-2-amine (YLIU-01-150-1)
[0602] To a mixture of compound YLIU-01-142-1 (70 mg, 0.14 mmol) in
MeCN (4 mL) was added TMSI (280 mg, 1.4 mmol) and the resulted
solution was stirred at 0.degree. C. for 30 min. The reaction was
quenched with water and the resulting solution was purified by
Prep-HPLC (MeOH/H.sub.2O, 0.05% TFA) to give the title compound (34
mg, 66%) as yellow solid. LCMS (m/z): 367 [M+H]+.
N-(4-((2R,3R)-3-((5-((5-tert-butyl)oxazol-2-yl)methyl)thio)triazol-2-yl)am-
ino)-2-methylpiperidine-1-carbonyl)phenyl)acrylamide
(YLIU-01-155-1)
[0603] To a mixture of YLIU-01-150-1 (17 mg, 0.046 mmol),
4-acrylamidobenzoic acid (10 mg, 0.055 mmol) and DIPEA (0.04 mL,
0.23 mmol) in DMF (2 mL) was added HATU (35 mg, 0.093 mmol) at rt.
The reaction mixture was stirred at rt overnight. The solution was
diluted with DCM (20 mL) and washed with brine (50 mL.times.2) and
dried over Na.sub.2SO.sub.4, filtered and concentrated. The residue
was purified by Prep-HPLC (MeOH/H.sub.2O, 0.05% TFA) to give the
title compound (8.2 mg, 33%) as yellow solid. LCMS (m/z): 540
[M+H].sup.+.
[0604] 1H NMR (500 MHz, DMSO) .delta. 10.30 (s, 1H), 8.02 (s, 1H),
7.72 (d, J=8.6 Hz, 2H), 7.38 (d, J=8.3 Hz, 2H), 6.79 (m, 2H), 6.45
(dd, J=17.0, 10.2 Hz, 1H), 6.28 (dd, J=17.0, 1.9 Hz, 1H), 5.79 (dd,
J=10.1, 1.9 Hz, 1H), 3.79 (m, 4H), 3.08 (m, 2H), 1.84-1.38 (m, 4H),
1.19 (s, 9H), 1.04 (s, 3H).
##STR00302##
benzyl
(2S,5R)-5-((5-((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl-
)amino)-2-methylpiperidine-1-carboxylate (YLIU-01-143-1)
[0605] A mixture of YLIU-01-017-1 (80 mg, 0.24 mmol), benzyl
(2S,5R)-5-amino-2-methyl piperidine-1-carboxylate (300 mg, 1.2
mmol), DIPEA (0.42 mL, 2.4 mmol) in NMP (3 mL) was heated to
140.degree. C. overnight. The solution was diluted with water (20
mL) and extracted with chloroform and iso-propanol (4:1). The
organic phase was washed with brine (50 mL.times.2) and dried over
Na.sub.2SO.sub.4, filtered and concentrated. The residue was
purified by silica gel column (MeOH/DCM, 0-20%) to give
YLIU-01-143-1 (50 mg, 42%) as yellow solid. LCMS (m/z): 501
[M+H]+.
5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)-N-((3R,6S)--6-methylpiperidin--
3-yl)triazol-2-amine (YLIU-01-151-1)
[0606] To a mixture of compound YLIU-01-143-1 (50 mg, 0.1 mmol) in
MeCN (4 mL) was added TMSI (200 mg, 1 mmol) and the resulted
solution was stirred at 0.degree. C. for 30 min. The reaction was
quenched with water and the resulting solution was purified by
Prep-HPLC (MeOH/H.sub.2O, 0.05% TFA) to give the title compound (20
mg, 54%) as yellow solid. LCMS (m/z): 367 [M+H]+.
N-(4-((2,5R)-5-((((5-(tertbutyl)oxazol-2-yl)methyl)thio)triazol-2-yl)amino-
)-2-methylpiperidine-1-carbonyl)phenyl)acrylamide
(YLIU-01-156-1)
[0607] To a mixture of YLIU-01-151-1 (20 mg, 0.054 mmol),
4-acrylamidobenzoic acid (12.5 mg, 0.065 mmol) and DIPEA (0.05 mL,
0.27 mmol) in DMF (2 mL) was added HATU (41 mg, 0.108 mmol) at rt.
The reaction mixture was stirred at rt overnight. The solution was
diluted with DCM (20 mL) and washed with brine (50 mL.times.2) and
dried over Na.sub.2SO.sub.4, filtered and concentrated. The residue
was purified by Prep-HPLC (MeOH/H2O, 0.05% TFA) to give the title
compound (11 mg, 38%) as yellow solid. LCMS (m/z): 540 [M+H]+. 1H
NMR (500 MHz, DMSO) .delta. 10.30 (s, 1H), 8.03 (s, 1H), 7.73 (d,
J=8.6 Hz, 2H), 7.39 (d, J=8.6 Hz, 2H), 6.97 (m, 1H), 6.71 (s, 1H),
6.45 (dd, J=17.0, 10.1 Hz, 1H), 6.28 (dd, J=17.0, 1.9 Hz, 1H), 5.79
(dd, J=10.1, 1.9 Hz, 1H), 3.95 (m, 2H), 3.57 (m, 2H), 3.34-3.02 (m,
1H), 2.68 (m, 1H), 1.71 (m, 4H), 1.35-0.88 (m, 12H).
Synthesis of B1
##STR00303## ##STR00304##
[0608] 5-thiocyanatothiazol-2-amine (1)
[0609] To a solution of 5-bromothiazol-2-amine hydrobromide (5.2 g,
20 mmol) in methanol (100 mL) was added KSCN (19.5 g, 200 mmol) at
room temperature. The resulting mixture was stirred for 20 hours
and then concentrated under vacuum. The residue was diluted with
H.sub.2O (150 mL). The pH of the solution was adjusted to 10 with
10% Na.sub.2CO.sub.3. The precipitate was filtered and washed with
water to obtain (1.8 g, 58%) of the title compound (1) as a brown
solid. MS m/z 158.95 [M+H].sup.+.
5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-amine (2)
[0610] To a solution of 5-thiocyanatothiazol-2-amine (1.8 g, 11.46
mmol) in absolute EtOH (50 mL) was added NaBH.sub.4 (0.87 g, 2.3
mmol) portionwise at room temperature. The resulting mixture was
stirred for 1 hour, and then acetone (20 ml) was slowly introduced.
After 1 hours, a solution of 5-(tert-butyl)-2-(chloromethyl)oxazole
(2.19 g, 12.6 mmol) in EtOH (10 ml) was added, and the resulting
mixture was cooled, concentrated in vacuo, and then partitioned
between EtOAc and brine, dried (Na.sub.2SO.sub.4), and
concentrated. The residue was purified by an silica gel column to
afford (2.1 g, 68%) of the title compound (2) as a pale red-brown
solid. MS m/z 270.06 [M+H].sup.+.
tert-butyl
4-((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)car-
bamoyl)piperidine-1-carboxylate (3)
[0611] To a solution of
5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-amine (186 mg,
0.69 mmol), 1-(tert-butoxycarbonyl)piperidine-4-carboxylic acid
(229 mg, 0.94 mmol), DMAP (42.8 mg, 0.35 mmol), and triethylamine
(0.2 ml, 1.4 mmol) in DMF (1 mL) and DCM (2 ml) was added EDAC (267
mg, 1.4 mmol) at room temperature. The reaction mixture was stirred
for 1.5 hours, diluted with EtOAc, washed with water and brine,
dried (Na.sub.2SO.sub.4), and concentrated. The residue was
purified by an silica gel column to afford (226 mg, 83%) of the
title compound (3) as a white solid. MS m/z 481.18[M+H]+.
N-(5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)piperidine-4-ca-
rboxamide hydrochloride (4)
[0612] To a solution of tert-butyl
4-((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)carbamoyl)pip-
eridine-1-carboxylate (117 mg, 0.24 mmol) in 4 M HCl EtOAc (5 mL).
The resulting mixture was stirred at room temperature for 30 min.
Then it was concentrated under vacuum to afford (100 mg, 100%) of
the title compound (4) as a white solid, which was used without
further purification. MS m/z 381.14 [M+H].sup.+.
(E)-N-(5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)-1-(4-(dime-
thylamino)but-2-enoyl)piperidine-4-carboxamide (B1)
[0613] To a solution of
N-(5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)piperidine-4-c-
arboxamide hydrochloride (100 mg, 0.24 mmol) in DMF (1 mL) was
added HATU (110 mg, 0.29 mmol), (E)-4-(dimethylamino)but-2-enoic
acid hydrochloride (45 mg, 0.27 mmol) and DIPEA (125 mg, 0.96
mmol). The resulting mixture was stirred at room temperature for 1
hour. Then it was diluted with EtOAc (100 mL), washed with water
and brine, dried (Na.sub.2SO.sub.4), and concentrated. The residue
was purified by an silica gel column to afford (6 mg, 5.1%) of the
title compound as a yellow solid. MS m/z 492.21 [M+H].sup.+.
.sup.1H NMR (400 MHz, DMSO-d.sub.6) 12.42 (s, 1H), 10.66 (s, 2H),
7.77 (s, 2H), 7.40 (s, 3H), 6.83 (s, 1H), 6.72 (s, 1H), 6.51 (d,
J=15.1 Hz, 1H), 4.52-4.17 (m, 1H), 4.07 (s, 2H), 3.63 (s, 1H), 3.08
(s, 2H), 2.77 (s, 6H), 2.02 (s, 2H), 1.72 (s, 3H), 1.46 (s, 2H),
1.19 (s, 9H).
Synthesis of B2
##STR00305## ##STR00306##
[0614] 5-thiocyanatothiazol-2-amine (1)
[0615] To a solution of 5-bromothiazol-2-amine hydrobromide (5.2 g,
20 mmol) in Methanol (100 mL) was added KSCN (19.5 g, 200 mmol) at
room temperature. The resulting mixture was stirred for 20 hours
and then concentrated under vacuum. The residue was diluted with
H.sub.2O (150 mL). The pH of the solution was adjusted to 10 with
10% Na.sub.2CO.sub.3. The precipitate was filtered and washed with
water to obtain (1.8 g, 58%) of the title compound (1) as a brown
solid. MS m/z 158.95 [M+H].sup.+.
5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-amine (2)
[0616] To a solution of 5-thiocyanatothiazol-2-amine (1.8 g, 11.46
mmol) in absolute EtOH (50 mL) was added NaBH.sub.4 (0.87 g, 2.3
mmol) portionwise at room temperature. The resulting mixture was
stirred for 1 hour, and then acetone (20 ml) was slowly introduced.
After 1 hour, a solution of 5-(tert-butyl)-2-(chloromethyl)oxazole
(2.19 g, 12.6 mmol) in EtOH (10 ml) was added, and the resulting
mixture was cooled, concentrated in vacuo, and then partitioned
between EtOAc and brine, dried (Na.sub.2SO.sub.4), and
concentrated. The residue was purified by an silica gel column to
afford (2.1 g, 68%) of the title compound (2) as a pale red-brown
solid. MS m/z 270.06 [M+H].sup.+.
tert-butyl
4-((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)car-
bamoyl)piperidine-1-carboxylate (3)
[0617] To a solution of
5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-amine (186 mg,
0.69 mmol), 1-(tert-butoxycarbonyl)piperidine-4-carboxylic acid
(229 mg, 0.94 mmol), DMAP (42.8 mg, 0.35 mmol), and triethylamine
(0.2 ml, 1.4 mmol) in DMF (1 mL) and DCM (2 ml) was added EDAC (267
mg, 1.4 mmol) at room temperature. The reaction mixture was stirred
for 1.5 hours, diluted with EtOAc, washed with water and brine,
dried (Na.sub.2SO.sub.4), and concentrated. The residue was
purified by an silica gel column to afford (226 mg, 83%) of the
title compound (3) as a white solid. MS m/z 481.18[M+H].sup.+.
N-(5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)piperidine-4-ca-
rboxamide hydrochloride (4)
[0618] To a solution of tert-butyl
4-((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)carbamoyl)pip-
eridine-1-carboxylate (20 mg, 0.047 mmol) in 4 M HCl EtOAc (3 mL).
The resulting mixture was stirred at room temperature for 30 min.
Then it was concentrated under vacuum to afford (100 mg, 100%) of
the title compound (4) as a white solid, which was used without
further purification. MS m/z 381.14 [M+H]+.
1-acryloyl-N-(5-(((5-(tert-butyl)oxazol-2-yl)methylthio)thiazol-2-yl)piper-
idine-4-carboxamide (B2)
[0619] To a solution of
N-(5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)piperidine-4-c-
arboxamide hydrochloride (20 mg, 0.05 mmol) in THF (5 mL) was added
DIPEA (17 mg, 0.15 mmol) and acryloyl chloride (43 mg, 0.05 mmol)
at 0.degree. C. for 1 hour. The resulting mixture was stirred at
room temperature for 2 hours. Then it was concentrated and purified
by an silica gel column to afford (11 mg, 50.7%) of the title
compound as a slightly white solid. MS m/z 435.15 [M+H].sup.+.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 11.64 (s, 1H), 7.29 (s,
1H), 6.58 (q, J=11.0 Hz, 2H), 6.28 (d, J=16.7 Hz, 1H), 5.71 (d,
J=10.6 Hz, 1H), 4.58 (d, J=12.1 Hz, 1H), 4.09 (dd, J=17.6, 10.2 Hz,
1H), 3.96 (s, 2H), 3.21 (s, 1H), 2.90 (s, 1H), 2.68 (t, J=10.4 Hz,
1H), 1.95 (d, J=12.0 Hz, 2H), 1.82 (s, 2H), 1.24 (s, 9H).
Synthesis of B3
##STR00307## ##STR00308##
[0620] 5-thiocyanatothiazol-2-amine (1)
[0621] To a solution of 5-bromothiazol-2-amine hydrobromide (5.2 g,
20 mmol) in methanol (100 mL) was added KSCN (19.5 g, 200 mmol) at
room temperature. The resulting mixture was stirred for 20 hours
and then concentrated under vacuum. The residue was diluted with
H.sub.2O (150 mL). The pH of the solution was adjusted to 10 with
10% Na.sub.2CO.sub.3. The precipitate was filtered and washed with
water to obtain (1.8 g, 58%) of the title compound (1) as a brown
solid. MS m/z 158.95 [M+H].sup.+.
5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-amine (2)
[0622] To a solution of 5-thiocyanatothiazol-2-amine (1.8 g, 11.46
mmol) in absolute EtOH (50 mL) was added NaBH.sub.4 (0.87 g, 2.3
mmol) portionwise at room temperature. The resulting mixture was
stirred for 1 hour, and then acetone (20 m) was slowly introduced.
After 1 hour, a solution of 5-(tert-butyl)-2-(chloromethyl)oxazole
(2.19 g, 12.6 mmol) in EtOH (10 ml) was added, and the resulting
mixture was cooled, concentrated in vacuo, and then partitioned
between EtOAc and brine, dried (Na.sub.2SO.sub.4), and
concentrated. The residue was purified by an silica gel column to
afford (2.1 g, 68%) of the title compound (2) as a pale red-brown
solid. MS m/z 270.06 [M+H].sup.+.
tert-butyl
(3-((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)triazol-2-yl)ca-
rbamoyl)cyclohexyl)carbamate (3)
[0623] To a solution of
5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-amine (160 mg,
0.6 mmol) in DMF (2 mL) was added HATU (456 mg, 1.2 mmol),
3-((tert-butoxycarbonyl)amino)cyclohexane-1-carboxylic acid (292
mg, 1.2 mmol), and DIPEA (143 mg, 2.4 mmol). The resulting mixture
was stirred at room temperature for 1 hour. Then it was diluted
with EtOAc (150 mL), washed with water and brine, dried
(Na.sub.2SO.sub.4), and concentrated. The residue was purified by
an silica gel column to afford (210 mg, 70%) of the title compound
(3) as a white solid. MS m/z 495.20[M+H].sup.+.
3-amino-N-(5-(((5-tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)cyclohex-
ane-1-carboxamide hydrochloride (4)
[0624] To a solution of tert-butyl
(3-((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)carbamoyl)cy-
clohexyl)carbamate (210 mg, 0.424 mmol) in 4 M HCl EtOAc (5 mL).
The resulting mixture was stirred at room temperature for 30 min.
Then it was concentrated under vacuum to afford (182 mg, 99%) of
the title compound (4) as a white solid, which was used without
further purification. MS m/z 395.15 [M+H].sup.+.
(E)-N-(5-((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)-3-(4-(dimet-
hylamino)but-2-enamido)cyclohexane-1-carboxamide (B3)
[0625] To a solution of
3-amino-N-(5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)cycloh-
exane-1-carboxamide hydrochloride (130 mg, 0.3 mmol) in DMF (1 mL)
was added HATU (228 mg, 0.6 mmol), (E)-4-(dimethylamino)but-2-enoic
acid hydrochloride (100 mg, 0.6 mmol), and DIPEA (0.214 ml, 1.2
mmol). The resulting mixture was stirred at room temperature for 1
hour. Then it was diluted with EtOAc (100 mL), washed with water
and brine, dried (Na.sub.2SO.sub.4), and concentrated. The residue
was purified by an silica gel column to afford (48 mg, 33%) of the
title compound as a white solid. MS m/z 506.26[M+H].sup.+. .sup.1H
NMR (400 MHz, DMSO-d.sub.6) .delta. 12.27 (s, 1H), 8.00 (d, J=8.0
Hz, 1H), 7.39 (s, 1H), 6.73 (s, 1H), 6.54 (d, J=16.2 Hz, 1H), 6.01
(d, J=15.7 Hz, 1H), 4.06 (s, 2H), 3.63 (d, J=26.0 Hz, 2H), 3.03 (s,
2H), 2.60 (s, 1H), 2.17 (s, 6H), 1.91 (d, J=11.2 Hz, 1H), 1.79 (s,
3H), 1.30 (s, 4H), 1.18 (s, 9H).
Synthesis of B4
##STR00309##
[0626] 5-thiocyanatothiazol-2-amine (1)
[0627] To a solution of 5-bromothiazol-2-amine hydrobromide (5.2 g,
20 mmol) in methanol (100 mL) was added KSCN (19.5 g, 200 mmol) at
room temperature. The resulting mixture was stirred for 20 hours
and then concentrated under vacuum. The residue was diluted with
H.sub.2O (150 mL). The pH of the solution was adjusted to 10 with
10% Na.sub.2CO.sub.3. The precipitate was filtered and washed with
water to obtain (1.8 g, 58%) of the title compound (1) as a brown
solid. MS m/z 158.95 [M+H].sup.+.
5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-amine (2)
[0628] To a solution of 5-thiocyanatothiazol-2-amine (1.8 g, 11.46
mmol) in absolute EtOH (50 mL) was added NaBH.sub.4 (0.87 g, 2.3
mmol) portionwise at room temperature. The resulting mixture was
stirred for 1 hour, and then acetone (20 ml) was slowly introduced.
After 1 hour, a solution of 5-(tert-butyl)-2-(chloromethyl)oxazole
(2.19 g, 12.6 mmol) in EtOH (10 ml) was added, and the resulting
mixture was cooled, concentrated in vacuo, and then partitioned
between EtOAc and brine, dried (Na.sub.2SO.sub.4), and
concentrated. The residue was purified by an silica gel column to
afford (2.1 g, 68%) of the title compound (2) as a pale red-brown
solid. MS m/z 270.06 [M+H].sup.+.
tert-butyl
3-((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)car-
bamoyl)piperidine-1-carboxylate (3)
[0629] To a solution of
5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-amine (200 mg,
0.74 mmol) in DMF (2 mL) was added HATU (562 mg, 1.5 mmol),
1-(tert-butoxycarbonyl)piperidine-3-carboxylic acid (343 mg, 1.5
mmol), and DIPEA (0.54 ml, 3 mmol). The resulting mixture was
stirred at room temperature for 1 hour. Then it was diluted with
EtOAc (150 mL), washed with water and brine, dried
(Na.sub.2SO.sub.4), and concentrated. The residue was purified by
an silica gel column to afford (260 mg, 73%) of the title compound
(3) as a white solid. MS m/z 481.19[M+H].sup.+.
N-(5-(((5-tert-butyl)oxazol-2-yl)methyl)thio)triazol-2-yl)piperidine-3-car-
boxamide hydrochloride (4)
[0630] To a solution of tert-butyl
3-((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)carbamoyl)pip-
eridine-1-carboxylate (260 mg, 0.54 mmol) in 4 M HCl EtOAc (5 mL).
The resulting mixture was stirred at room temperature for 30 min.
Then it was concentrated under vacuum to afford (180 mg, 80%) of
the title compound (4) as a white solid, which was used without
further purification. MS m/z 381.14 [M+H].sup.+.
tert-butyl
(4-(3-((5-((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)-
carbamoyl)piperidine-1-carbonyl)phenyl)carbamate (5)
[0631] To a solution of
N-(5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)piperidine-3-c-
arboxamide hydrochloride (180 mg, 0.43 mmol) in DMF (2 mL) was
added HATU (329 mg, 0.86 mmol),
4-((tert-butoxycarbonyl)amino)benzoic acid (205 mg, 0.86 mmol), and
DIPEA (0.38 ml, 2.15 mmol). The resulting mixture was stirred at
room temperature for 1 hour. Then it was diluted with EtOAc (100
mL), washed with water and brine, dried (Na.sub.2SO.sub.4), and
concentrated. The residue was purified by an silica gel column to
afford (80 mg, 31%) of the title compound (5) as a white solid. MS
m/z 600.23[M+H].sup.+.
1-(4-aminobenzoyl)-N-(5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-
-yl)piperidine-3-carboxamide hydrochloride (6)
[0632] To a solution of tert-butyl
(4-(3-((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)carbamoyl-
)piperidine-1-carbonyl)phenyl)carbamate (80 mg, 0.133 mmol) in 4 M
HCl EtOAc (3 mL). The resulting mixture was stirred at room
temperature for 30 min. Then it was concentrated under vacuum to
afford (65 mg, 91%) of the title compound (6) as a white solid,
which was used without further purification. MS m/z 381.14
[M+H].sup.+.
(E)-N-(5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)-1-(4-(4-(d-
imethylamino)but-2-enamido)benzoyl)piperidine-3-carboxamide
(B4)
[0633] To a solution of
1-(4-aminobenzoyl)-N-(5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol--
2-yl)piperidine-3-carboxamide hydrochloride (0.60 g, 0.112 mmol) in
DCM (5 mL) was added DIPEA (0.2 ml, 1.34 mmol) and
(E)-4-bromobut-2-enoyl chloride (22.5 mg, 0.134 mmol) at 0.degree.
C. for 3 min. Then it was added 4.0 M dimethylamine in THF (2 mL).
The resulting mixture was stirred at room temperature for 2 hours.
Then it was concentrated and purified by an silica gel column to
afford (8 mg, 11.7%) of the title compound as a slightly white
solid. MS m/z 611.24 [M+H].sup.+. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) .delta. 12.42 (s, 1H), 10.66 (s, 2H), 7.77 (s, 2H),
7.40 (s, 3H), 6.83 (s, 1H), 6.72 (s, 1H), 6.51 (d, J=15.1 Hz, 1H),
4.52-4.17 (m, 1H), 4.07 (s, 2H), 3.63 (s, 1H), 3.08 (s, 2H), 2.77
(s, 6H), 2.02 (s, 2H), 1.72 (s, 3H), 1.46 (s, 2H), 1.19 (s,
9H).
Synthesis of B5
##STR00310## ##STR00311##
[0634] 5-thiocyanatothiazol-2-amine (1)
[0635] To a solution of 5-bromothiazol-2-amine hydrobromide (5.2 g,
20 mmol) in methanol (100 mL) was added KSCN (19.5 g, 200 mmol) at
room temperature. The resulting mixture was stirred for 20 hours
and then concentrated under vacuum. The residue was diluted with
H.sub.2O (150 mL). The pH of the solution was adjusted to 10 with
10% Na.sub.2CO.sub.3. The precipitate was filtered and washed with
water to obtain (1.8 g, 58%) of the title compound (1) as a brown
solid. MS m/z 158.95 [M+H].sup.+.
5-((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-amine (2)
[0636] To a solution of 5-thiocyanatothiazol-2-amine (1.8 g, 11.46
mmol) in absolute EtOH (50 mL) was added NaBH.sub.4 (0.87 g, 2.3
mmol) portionwise at room temperature. The resulting mixture was
stirred for 1 hour, and then acetone (20 ml) was slowly introduced.
After 1 hour, a solution of 5-(tert-butyl)-2-(chloromethyl)oxazole
(2.19 g, 12.6 mmol) in EtOH (10 ml) was added, and the resulting
mixture was cooled, concentrated in vacuo, and then partitioned
between EtOAc and brine, dried (Na.sub.2SO.sub.4), and
concentrated. The residue was purified by an silica gel column to
afford (2.1 g, 68%) of the title compound (2) as a pale red-brown
solid. MS m/z 270.06 [M+H].sup.+.
tert-butyl
3-((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)car-
bamoyl)piperidine-1-carboxylate (3)
[0637] To a solution of
5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-amine (65 mg,
0.24 mmol) in DMF (2 mL) was added HATU (183 mg, 0.48 mmol),
1-(tert-butoxycarbonyl)piperidine-3-carboxylic acid (83 mg, 0.36
mmol) and DIPEA (126 mg, 0.96 mmol). The resulting mixture was
stirred at room temperature for 1 hour. Then it was diluted with
EtOAc (100 mL), washed with water and brine, dried
(Na.sub.2SO.sub.4), and concentrated. The residue was purified by
an silica gel column to afford (23 mg, 20%) of the title compound
(3) as a white solid. MS m/z 481.19[M+H].sup.+.
N-(5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)piperidine-3-ca-
rboxamide hydrochloride (4)
[0638] To a solution of tert-butyl
3-((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)carbamoyl)pip-
eridine-1-carboxylate (23 mg, 0.048 mmol) in 4 M HCl EtOAc (2 mL).
The resulting mixture was stirred at room temperature for 30 min.
Then it was concentrated under vacuum to afford (16 mg, 80%) of the
title compound (4) as a white solid, which was used without further
purification. MS m/z 381.14 [M+H]+.
1-(4-acrylamidobenzoyl)-N-(5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thia-
zol-2-yl)piperidine-3-carboxamide (B5)
[0639] To a solution of
N-(5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)piperidine-3-c-
arboxamide hydrochloride (50 mg, 0.12 mmol) in DMF (1 mL) was added
HATU (92 mg, 0.24 mmol), 4-acrylamidobenzoic acid (46 mg, 0.24
mmol), and DIPEA (0.22 ml, 1.2 mmol). The resulting mixture was
stirred at room temperature for 1 hour. Then it was diluted with
EtOAc (100 mL), washed with water and brine, dried
(Na.sub.2SO.sub.4), and concentrated. The residue was purified by
an silica gel column to afford (8 mg, 12%) of the title compound as
a white solid. MS m/z 554.21 [M+H].sup.+. .sup.1H NMR (400 MHz,
DMSO-ds) .delta. 12.37 (s, 1H), 7.73 (d, J=7.8 Hz, 2H), 7.55-7.22
(m, 3H), 6.71 (s, 1H), 6.44 (dd, J=16.9, 10.3 Hz, 1H), 6.28 (d,
J=16.8 Hz, 1H), 5.78 (d, J=9.9 Hz, 1H), 4.05 (s, 2H), 3.67 (s, 1H),
3.05 (s, 2H), 2.69 (s, 1H), 1.97 (s, 1H), 1.70 (d, J=12.1 Hz, 2H),
1.44 (s, 1H), 1.23 (s, 2H), 1.12 (s, 9H).
Synthesis of B6
##STR00312##
[0640] 5-thiocyanatothiazol-2-amine (1)
[0641] To a solution of 5-bromothiazol-2-amine hydrobromide (5.2 g,
20 mmol) in methanol (100 mL) was added KSCN (19.5 g, 200 mmol) at
room temperature. The resulting mixture was stirred for 20 hours
and then concentrated under vacuum. The residue was diluted with
H.sub.2O (150 mL). The pH of the solution was adjusted to 10 with
10% Na.sub.2CO.sub.3. The precipitate was filtered and washed with
water to obtain (1.8 g, 58%) of the title compound (1) as a brown
solid. MS m/z 158.95 [M+H].sup.+.
5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-amine (2)
[0642] To a solution of 5-thiocyanatothiazol-2-amine (1.8 g, 11.46
mmol) in absolute EtOH (50 mL) was added NaBH.sub.4 (0.87 g, 2.3
mmol) portionwise at room temperature. The resulting mixture was
stirred for 1 hour, and then acetone (20 ml) was slowly introduced.
After 1 hour, a solution of 5-(tert-butyl)-2-(chloromethyl)oxazole
(2.19 g, 12.6 mmol) in EtOH (10 ml) was added, and the resulting
mixture was cooled, concentrated in vacuo, and then partitioned
between EtOAc and brine, dried (Na.sub.2SO.sub.4), and
concentrated. The residue was purified by an silica gel column to
afford (2.1 g, 68%) of the title compound (2) as a pale red-brown
solid. MS m/z 270.06 [M+H].sup.+.
3-acrylamido-N-(5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)be-
nzamide (B6)
[0643] To a solution of
5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-amine (54 mg,
0.2 mmol) in DMF (1 mL) was added HATU (160 mg, 0.42 mmol),
3-acrylamidobenzoic acid (60 mg, 0.31 mmol), and DIPEA (52 mg, 0.4
mmol). The resulting mixture was stirred at room temperature for 1
hour. Then it was diluted with EtOAc (100 mL), washed with water
and brine, dried (Na.sub.2SO.sub.4), and concentrated. The residue
was purified by an silica gel column to afford (7 mg, 8%) of the
title compound as a white solid. MS m/z 443.12[M+H].sup.+. .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. 12.36 (s, 1H), 9.08 (s, 1H),
8.24-7.99 (m, 2H), 7.90 (s, 1H), 7.63 (d, J=7.6 Hz, 1H), 7.41 (t,
J=7.9 Hz, 1H), 6.95 (s, 1H), 6.55 (s, 1H), 6.44 (d, J=16.8 Hz, 1H),
6.32 (dd, J=16.8, 10.0 Hz, 1H), 5.76 (d, J=10.0 Hz, 1H), 3.95 (s,
2H), 1.23 (s, 9H).
Synthesis of B7
##STR00313##
[0644] 5-thiocyanatothiazol-2-amine (1)
[0645] To a solution of 5-bromothiazol-2-amine hydrobromide (5.2 g,
20 mmol) in methanol (100 mL) was added KSCN (19.5 g, 200 mmol) at
room temperature. The resulting mixture was stirred for 20 hours
and then concentrated under vacuum. The residue was diluted with
H.sub.2O (150 mL). The pH of the solution was adjusted to 10 with
10% Na.sub.2CO.sub.3. The precipitate was filtered and washed with
water to obtain (1.8 g, 58%) of the title compound (1) as a brown
solid. MS m/z 158.95 [M+H].sup.+.
5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-amine (2)
[0646] To a solution of 5-thiocyanatothiazol-2-amine (1.8 g, 11.46
mmol) in absolute EtOH (50 mL) was added NaBH.sub.4 (0.87 g, 2.3
mmol) portionwise at room temperature. The resulting mixture was
stirred for 1 hour, and then acetone (20 ml) was slowly introduced.
After 1 hour, a solution of 5-(tert-butyl)-2-(chloromethyl)oxazole
(2.19 g, 12.6 mmol) in EtOH (10 ml) was added, and the resulting
mixture was cooled, concentrated in vacuo, and then partitioned
between EtOAc and brine, dried (Na.sub.2SO.sub.4), and
concentrated. The residue was purified by an silica gel column to
afford (2.1 g, 68%) of the title compound (2) as a pale red-brown
solid. MS m/z 270.06 [M+H].sup.+.
tert-butyl
(3-((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)ca-
rbamoyl)phenyl)carbamate (3)
[0647] To a solution of
5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-amine (404 mg,
1.5 mmol) in DMF (2 mL) was added HATU (1.14 g, 3 mmol),
3-((tert-butoxycarbonyl)amino)benzoic acid (573 mg, 3 mmol), and
DIPEA (1.05 ml, 6 mmol). The resulting mixture was stirred at room
temperature for 1 hour. Then it was diluted with EtOAc (200 mL),
washed with water and brine, dried (Na.sub.2SO.sub.4), and
concentrated. The residue was purified by an silica gel column to
afford (490 mg, 67%) of the title compound (3) as a white solid. MS
m/z 489.15[M+H].sup.+.
3-amino-N-(5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)benzami-
de hydrochloride (4)
[0648] To a solution of tert-butyl
(3-((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)carbamoyl)ph-
enyl)carbamate (245 mg, 0.5 mmol) in 4 M HCl EtOAc (5 mL). The
resulting mixture was stirred at room temperature for 30 min. Then
it was concentrated under vacuum to afford (212 mg, 100%) of the
title compound (4) as a white solid, which was used without further
purification. MS m/z 389.11 [M+H].sup.+.
(E)-N-(5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)-3-(4-(dime-
thylamino)but-2-enamido)benzamide (B7)
[0649] To a solution of
3-amino-N-(5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)benzam-
ide hydrochloride (83 mg, 0.195 mmol) in MeCN (5 mL) was added
DIPEA (77.6 mg, 0.6 mmol), and (E)-4-bromobut-2-enoyl chloride
(40.26 mg, 0.22 mmol) at 0.degree. C. for 3 min. Then it was added
4.0 M dimethylamine in THF (1 mL). The resulting mixture was
stirred at room temperature for 2 hours. Then it was concentrated
and purified by an silica gel column to afford (7 mg, 7.2%) of the
title compound as a slightly yellow solid. MS m/z 500.18
[M+H].sup.+. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.82 (s,
1H), 8.01 (d, J=16.7 Hz, 2H), 7.64 (s, 1H), 7.42 (s, 1H), 7.10 (s,
1H), 7.00 (s, 1H), 6.58 (s, 1H), 6.24 (d, J=13.7 Hz, 1H), 3.96 (s,
2H), 3.16 (s, 2H), 2.31 (s, 6H), 1.26 (s, 9H).
Synthesis of B8
##STR00314##
[0650] 5-thiocyanatothiazol-2-amine (1)
[0651] To a solution of 5-bromothiazol-2-amine hydrobromide (5.2 g,
20 mmol) in methanol (100 mL) was added KSCN (19.5 g, 200 mmol) at
room temperature. The resulting mixture was stirred for 20 h and
then concentrated under vacuum. The residue was diluted with
H.sub.2O (150 mL). The pH of the solution was adjusted to 10 with
10% Na.sub.2CO.sub.3. The precipitate was filtered and washed with
water to obtain (1.8 g, 58%) of the title compound (1) as a brown
solid. MS m/z 158.95 [M+H]*.
5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-amine (2)
[0652] To a solution of 5-thiocyanatothiazol-2-amine (1.8 g, 11.46
mmol) in absolute EtOH (50 mL) was added NaBH.sub.4 (0.87 g, 2.3
mmol) portionwise at room temperature. The resulting mixture was
stirred for 1 hour, and then acetone (20 mL) was slowly introduced.
After 1 hour, a solution of 5-(tert-butyl)-2-(chloromethyl)oxazole
(2.19 g, 12.6 mmol) in EtOH (10 mL) was added, and the resulting
mixture was cooled, concentrated in vacuo, and then partitioned
between EtOAc and brine, dried (Na.sub.2SO.sub.4), and
concentrated. The residue was purified by an silica gel column to
afford (2.1 g, 68%) of the title compound (2) as a pale red-brown
solid. MS m/z 270.06 [M+H].sup.+.
tert-butyl
(4-((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)ca-
rbamoyl)phenyl)carbamate (3)
[0653] To a solution of
5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-amine (220 mg,
0.82 mmol) in DMF (1 mL) was added HATU (623 mg, 1.64 mmol),
4-((tert-butoxycarbonyl)amino)benzoic acid (390 mg, 1.64 mmol) and
DIPEA (0.6 ml, 3.28 mmol). The resulting mixture was stirred at
room temperature for 1 hour. Then it was diluted with EtOAc (150
mL), washed with water and brine, dried (Na.sub.2SO.sub.4), and
concentrated. The residue was purified by an silica gel column to
afford (200 mg, 50%) of the title compound (3) as a white solid. MS
m/z 489.15[M+H].sup.+.
4-amino-N-(5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)benzami-
de hydrochloride (4)
[0654] To a solution of tert-butyl
(4-((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)carbamoyl)ph-
enyl)carbamate (200 mg, 0.41 mmol) in 4 M HCl EtOAc (5 mL). The
resulting mixture was stirred at room temperature for 30 min. Then
it was concentrated under vacuum to afford (174 mg, 100%) of the
title compound (4) as a white solid, which was used without further
purification. MS m/z 389.11 [M+H].sup.+.
(E)-N-(5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)-44-(dimeth-
ylamino)but-2-enamido)benzamide (B8)
[0655] To a solution of
4-amino-N-(5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)benzam-
ide hydrochloride (75 mg, 0.177 mmol) in MeCN (5 mL) was added
DIPEA (0.3 ml, 2.6 mmol) and (E)-4-bromobut-2-enoyl chloride (46.85
mg, 0.256 mmol) at 0.degree. C. for 3 min. Then it was added 4.0 M
dimethylamine in THF (1 mL). The resulting mixture was stirred at
room temperature for 2 hours. Then it was concentrated and purified
by an silica gel column to afford (7 mg, 8%) of the title compound
as a slightly yellow solid. MS m/z 500.18 [M+H].sup.+. .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 8.57 (s, 1H), 7.90 (d, J=7.3 Hz, 2H),
7.76 (s, 2H), 7.19 (s, 1H), 7.05 (s, 1H), 6.61 (s, 1H), 6.23 (d,
J=15.5 Hz, 1H), 3.98 (s, 2H), 3.18 (s, 2H), 2.34 (s, 6H), 1.27 (s,
9H).
Synthesis of B9
##STR00315##
[0656] 5-thiocyanatothiazol-2-amine (1)
[0657] To a solution of 5-bromothiazol-2-amine hydrobromide (5.2 g,
20 mmol) in methanol (100 mL) was added KSCN (19.5 g, 200 mmol) at
room temperature. The resulting mixture was stirred for 20 h and
then concentrated under vacuum. The residue was diluted with
H.sub.2O (150 mL). The pH of the solution was adjusted to 10 with
10% Na.sub.2CO.sub.3. The precipitate was filtered and washed with
water to obtain (1.8 g, 58%) of the title compound (1) as a brown
solid. MS m/z 158.95 [M+H].sup.+.
5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-amine (2)
[0658] To a solution of 5-thiocyanatothiazol-2-amine (1.8 g, 11.46
mmol) in absolute EtOH (50 mL) was added NaBH.sub.4 (0.87 g, 2.3
mmol) portionwise at room temperature. The resulting mixture was
stirred for 1 hour, and then acetone (20 mL) was slowly introduced.
After 1 hour, a solution of 5-(tert-butyl)-2-(chloromethyl)oxazole
(2.19 g, 12.6 mmol) in EtOH (10 mL) was added, and the resulting
mixture was cooled, concentrated in vacuo, and then partitioned
between EtOAc and brine, dried (Na.sub.2SO.sub.4), and
concentrated. The residue was purified by an silica gel column to
afford (2.1 g, 68%) of the title compound (2) as a pale red-brown
solid. MS m/z 270.06 [M+H].sup.+.
tert-butyl
(3-((5-(((5-(tert-butyl)oxazol-2-yl)methyl))thio)thiazol-2-yl)c-
arbamoyl)phenyl)carbamate (3)
[0659] To a solution of
5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-amine (404 mg,
1.5 mmol) in DMF (2 mL) was added HATU (1.14 g, 3 mmol),
3-((tert-butoxycarbonyl)amino)benzoic acid (573 mg, 3 mmol), and
DIPEA (1.05 ml, 6 mmol). The resulting mixture was stirred at room
temperature for 1 hour. Then it was diluted with EtOAc (200 mL),
washed with water and brine, dried (Na.sub.2SO.sub.4), and
concentrated. The residue was purified by an silica gel column to
afford (490 mg, 67%) of the title compound (3) as a white solid. MS
m/z 489.15 [M+H].sup.+.
3-amino-N-(5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)benzami-
de hydrochloride (4)
[0660] To a solution of tert-butyl
(3-((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)carbamoyl)ph-
enyl)carbamate (245 mg, 0.5 mmol) in 4 M HCl EtOAc (5 mL). The
resulting mixture was stirred at room temperature for 30 min. Then
it was concentrated under vacuum to afford (212 mg, 100%) of the
title compound (4) as a white solid, which was used without further
purification. MS m/z 389.11 [M+H].sup.+.
3-(4-acrylamidobenzamido)-N-(5-(((5-tert-butyl)oxazol-2-yl)methyl)thio)tri-
azol-2-yl)benzamide (B9)
[0661] To a solution of
3-amino-N-(5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)benzam-
ide hydrochloride (212 mg, 0.5 mmol) in DMF (2 mL) was added HATU
(380 mg, 1 mmol), 4-acrylamidobenzoic acid (191 mg, 1 mmol), and
DIPEA (0.5 ml, 3 mmol). The resulting mixture was stirred at room
temperature for 1 hour. Then it was diluted with EtOAc (200 mL),
washed with water and brine, dried (Na.sub.2SO.sub.4), and
concentrated. The residue was purified by an silica gel column to
afford (93 mg, 34%) of the title compound as a white solid. MS m/z
562.18[M+H].sup.+. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.
12.83 (s, 1H), 10.47 (s, 1H), 10.40 (s, 1H), 8.46 (s, 1H), 8.00
(dd, J=15, 5.0 Hz, 3H), 7.83 (d, J=15.0 Hz, 3H), 7.52 (d, J=5.0 Hz,
2H), 6.75 (s, 1H), 6.46 (d, J=6.3 Hz, 1H), 6.31 (d, J=6.5 Hz, 1H),
5.82 (d, J=15.0 Hz, 1H), 4.12 (s, 2H), 1.22 (s, 9H).
Synthesis of B12
##STR00316##
[0662] 5-thiocyanatothiazol-2-amine (1)
[0663] To a solution of 5-bromothiazol-2-amine hydrobromide (26 g,
100 mmol) in methanol (500 mL) was added KSCN (50 g, 500 mmol) at
room temperature. The resulting mixture was stirred for 20 hours
and then concentrated under vacuum. The residue was diluted with
H.sub.2O (300 mL). The pH of the solution was adjusted to 10 with
10% Na.sub.2CO.sub.3. The precipitate was filtered and washed with
water to obtain (8 g, 51%) of the title compound (1) as a brown
solid. MS m/z 158.95 [M+H].sup.+.
5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-amine (2)
[0664] To a solution of 5-thiocyanatothiazol-2-amine (2 g, 13 mmol)
in absolute EtOH (50 mL) was added NaBH.sub.4 (1 g, 26 mmol)
portionwise at room temperature. The resulting mixture was stirred
for 1 hour, and then acetone (20 mL) was slowly introduced. After 1
hour, a solution of 5-(tert-butyl)-2-(chloromethyl)oxazole (2.71 g,
15.6 mmol) in EtOH (10 mL) was added, and the resulting mixture was
cooled, concentrated in vacuo, and then partitioned between EtOAc
and brine, dried (Na.sub.2SO.sub.4), and concentrated. The residue
was purified by an silica gel column to afford (2.8 g, 80%) of the
title compound (2) as a pale red-brown solid. MS m/z 270.06
[M+H].sup.+.
2-(((2-bromothiazol-5-yl)thio)methyl)-5-(tert-butyl)oxazole (3)
[0665] To a solution of
5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-amine (404 mg,
1.5 mmol) in MeCN (10 mL) was added tert-butyl nitrite (239 mg,
2.25 mmol) at 0.degree. C. The resulting mixture was stirred for 10
min, and then CuBr.sub.2 (402 mg, 1.8 mmol) was introduced. The
resulting mixture was stirred at room temperature for 3 hours, the
resulting mixture was concentrated in vacuo. The residue was
purified by an silica gel column to afford (143 mg, 29%) of the
title compound (3) as a pale yellow oil. MS m/z 333.96
[M+H].sup.+.
tert-butyl
3-((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)ami-
no)piperidine-1-carboxylate (4)
[0666] To a solution of
2-(((2-bromothiazol-5-yl)thio)methyl)-5-(tert-butyl)oxazole (80 mg,
0.22 mmol) in NMP (0.1 mL) was added tert-butyl
3-aminopiperidine-1-carboxylate (80 mg, 0.4 mmol) and DIPEA (0.084
ml, 0.48 mmol). The resulting mixture was heated at 140.degree. C.
for 8 hours. Then it was diluted with EtOAc (150 mL) at room
temperature, washed with water and brine, dried (Na.sub.2SO.sub.4),
and concentrated. The residue was purified by an silica gel column
to afford (30 mg, 28%) of the title compound (4) as a slightly
yellow solid. MS m/z 453.20 [M+H].sup.+.
5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)-N-(piperidin-3-yl)thiazol-2-am-
ine hydrochloride (5)
[0667] To a solution of tert-butyl
3-((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)amino)piperid-
ine-1-carboxylate (38 mg, 0.084 mmol) in 4 M HCl EtOAc (5 mL). The
resulting mixture was stirred at room temperature for 30 min. Then
it was concentrated under vacuum to afford (35 mg, 98%) of the
title compound (5) as a white solid, which was used without further
purification. MS m/z 353.14 [M+H].sup.+.
N-(4-(3-((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)amino)pi-
peridine-1-carbonyl)phenyl)acrylamide (B12)
[0668] To a solution of
5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)-N-(piperidin-3-yl)thiazol-2-a-
mine hydrochloride (35 mg, 0.082 mmol) in DMF (0.5 mL) was added
HATU (62.4 mg, 0.164 mmol), 4-acrylamidobenzoic acid (31.5 mg,
0.164 mmol), and DIPEA (42.6 mg, 0.33 mmol). The resulting mixture
was stirred at room temperature for 1 hour. Then it was diluted
with EtOAc (70 mL), washed with water and brine, dried
(Na.sub.2SO.sub.4), and concentrated. The residue was purified by
an silica gel column to afford (5 mg, 12%) of the title compound as
a white solid. MS m/z 526.21[M+H].sup.+. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 9.83 (s, 1H), 7.59 (d, J=7.2 Hz, 2H), 7.29 (s,
2H), 6.86 (s, 1H), 6.66 (s, 1H), 6.56 (s, 1H), 6.44 (d, J=16.9 Hz,
1H), 6.26 (s, 1H), 5.72 (s, 1H), 4.03 (s, 1H), 3.84 (s, 2H), 3.55
(s, 1H), 3.46 (d, J=31.2 Hz, 2H), 2.03 (s, 3H), 1.84 (s, 2H), 1.32
(s, 9H).
Synthesis of B15
##STR00317##
[0669] 5-thiocyanatothiazol-2-amine (1)
[0670] To a solution of 5-bromothiazol-2-amine hydrobromide (26 g,
100 mmol) in methanol (500 mL) was added KSCN (50 g, 500 mmol) at
room temperature. The resulting mixture was stirred for 20 h and
then concentrated under vacuum. The residue was diluted with
H.sub.2O (300 mL). The pH of the solution was adjusted to 10 with
10% Na.sub.2CO.sub.3. The precipitate was filtered and washed with
water to obtain (8 g, 51%) of the title compound (1) as a brown
solid. MS m/z 158.95 [M+H].sup.+.
5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-amine (2)
[0671] To a solution of 5-thiocyanatothiazol-2-amine (2 g, 13 mmol)
in absolute EtOH (50 mL) was added NaBH.sub.4 (1 g, 26 mmol)
portionwise at room temperature. The resulting mixture was stirred
for 1 hour, and then acetone (20 mL) was slowly introduced. After 1
hour, a solution of 5-(tert-butyl)-2-(chloromethyl)oxazole (2.71 g,
15.6 mmol) in EtOH (10 mL) was added, and the resulting mixture was
cooled, concentrated in vacuo, and then partitioned between EtOAc
and brine, dried (Na.sub.2SO.sub.4), and concentrated. The residue
was purified by an silica gel column to afford (2.8 g, 80%) of the
title compound (2) as a pale red-brown solid. MS m/z 270.06
[M+H].sup.+.
2-(((2-bromothiazol-5-yl)thio)methyl)-5-(tert-butyl)oxazole (3)
[0672] To a solution of
5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-amine (2.02 g,
7.5 mmol) in MeCN (50 mL) was added tert-butyl nitrite (1.2 g, 11.7
mmol) at 0.degree. C. The resulting mixture was stirred for 10 min,
and then CuBr.sub.2 (2.01 mg, 9 mmol) was introduced. The resulting
mixture was stirred at room temperature for 3 hours. the resulting
mixture was concentrated in vacuo. The residue was purified by an
silica gel column to afford (0.6 g, 24%) of the title compound (3)
as a pale yellow oil. MS m/z 333.96 [M+H].sup.+.
tert-butyl
3-((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)ami-
no)piperidine-1-carboxylate (4)
[0673] To a solution of
2-(((2-bromothiazol-5-yl)thio)methyl)-5-(tert-butyl)oxazole (0.6 g,
1.8 mmol) in NMP (0.6 mL) was added tert-butyl
3-aminopiperidine-1-carboxylate (0.72 mg, 3.6 mmol) and DIPEA (0.65
ml, 3.6 mmol). The resulting mixture was heated at 140.degree. C.
for 8 hours. Then it was diluted with EtOAc (200 mL) at room
temperature, washed with water and brine, dried (Na.sub.2SO.sub.4),
and concentrated. The residue was purified by an silica gel column
to afford (150 mg, 18.5%) of the title compound (4) as a slightly
yellow solid. MS m/z 453.20 [M+H].sup.+.
5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)-N-(piperidin-3-yl)triazol-2-am-
ine hydrochloride (5)
[0674] To a solution of tert-butyl
3-((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)amino)piperid-
ine-1-carboxylate (150 mg, 0.33 mmol) in 4 M HCl EtOAc (5 mL). The
resulting mixture was stirred at room temperature for 30 min. Then
it was concentrated under vacuum to afford (130 mg, 93%) of the
title compound (5) as a white solid, which was used without further
purification. MS m/z 353.14 [M+H].sup.+.
N-(3-(3-((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)triazol-2-yl)amino)pi-
peridine-1-carbonyl)phenyl)acrylamide (B12)
[0675] To a solution of
5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)-N-(piperidin-3-yl)thiazol-2-a-
mine hydrochloride (55 mg, 0.13 mmol) in DMF (0.5 mL) was added
HATU (99 mg, 0.26 mmol), 3-acrylamidobenzoic acid (49.7 mg, 0.26
mmol), and DIPEA (67 mg, 0.52 mmol). The resulting mixture was
stirred at room temperature for 1 hour. Then it was diluted with
EtOAc (70 mL), washed with water and brine, dried
(Na.sub.2SO.sub.4), and concentrated. The residue was purified by
an silica gel column to afford (12 mg, 18%) of the title compound
as a white solid. MS m/z 526.21 [M+H].sup.+. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 9.41 (s, 1H), 7.85 (s, 1H), 7.57 (s, 1H), 7.07
(s, 1H), 6.86 (s, 1H), 6.64 (s, 1H), 6.45 (d, J=17.1 Hz, 1H),
6.41-6.20 (m, 1H), 5.73 (s, 1H), 5.55 (s, 1H), 4.03-3.80 (m, 2H),
3.71 (s, 1H), 3.56 (s, 2H), 2.04 (s, 2H), 1.85 (s, 4H), 1.29 (s,
9H).
Synthesis of B16
##STR00318##
[0676] 5-thiocyanatothiazol-2-amine (1)
[0677] To a solution of 5-bromothiazol-2-amine hydrobromide (26 g,
100 mmol) in methanol (500 mL) was added KSCN (50 g, 500 mmol) at
room temperature. The resulting mixture was stirred for 20 hours
and then concentrated under vacuum. The residue was diluted with
H.sub.2O (300 mL). The pH of the solution was adjusted to 10 with
10% Na.sub.2CO.sub.3. The precipitate was filtered and washed with
water to obtain (8 g, 51%) of the title compound (1) as a brown
solid. MS m/z 158.95 [M+H].sup.+.
5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-amine (2)
[0678] To a solution of 5-thiocyanatothiazol-2-amine (2 g, 13 mmol)
in absolute EtOH (50 mL) was added NaBH.sub.4 (1 g, 26 mmol)
portionwise at room temperature. The resulting mixture was stirred
for 1 hour, and then acetone (20 mL) was slowly introduced. After 1
hour, a solution of 5-(tert-butyl)-2-(chloromethyl)oxazole (2.71 g,
15.6 mmol) in EtOH (10 mL) was added, and the resulting mixture was
cooled, concentrated in vacuo, and then partitioned between EtOAc
and brine, dried (Na.sub.2SO.sub.4), and concentrated. The residue
was purified by an silica gel column to afford (2.8 g, 80%) of the
title compound (2) as a pale red-brown solid. MS m/z 270.06
[M+H].sup.+.
2-(((2-bromothiazol-5-yl)thio)methyl)-5-(tert-butyl)oxazole (3)
[0679] To a solution of
5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-amine (2.02 g,
7.5 mmol) in MeCN (50 mL) was added tert-butyl nitrite (1.2 g, 11.7
mmol) at 0.degree. C. The resulting mixture was stirred for 10 min,
and then CuBr.sub.2 (2.01 mg, 9 mmol) was introduced. The resulting
mixture was stirred at room temperature for 3 hours. the resulting
mixture was concentrated in vacuo. The residue was purified by an
silica gel column to afford (0.6 g, 24%) of the title compound (3)
as a pale yellow oil. MS m/z 333.96 [M+H].sup.+.
tert-butyl
3-((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)ami-
no)piperidine-1-carboxylate (4)
[0680] To a solution of
2-(((2-bromothiazol-5-yl)thio)methyl)-5-(tert-butyl)oxazole (0.6 g,
1.8 mmol) in NMP (0.6 mL) was added tert-butyl
3-aminopiperidine-1-carboxylate (0.72 mg, 3.6 mmol) and DIPEA (0.65
ml, 3.6 mmol). The resulting mixture was heated at 140.degree. C.
for 8 hours. Then it was diluted with EtOAc (200 mL) at room
temperature, washed with water and brine, dried (Na.sub.2SO.sub.4),
and concentrated. The residue was purified by an silica gel column
to afford (150 mg, 18.5%) of the title compound (4) as a slightly
yellow solid. MS m/z 453.20 [M+H].sup.+.
5-((5-(tert-butyl)oxazol-2-yl)methyl)thio)-N-(piperidin-3-yl)thiazol-2-ami-
ne hydrochloride (5)
[0681] To a solution of tert-butyl
3-((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)amino)piperid-
ine-1-carboxylate (150 mg, 0.33 mmol) in 4 M HCl EtOAc (5 mL). The
resulting mixture was stirred at room temperature for 30 min. Then
it was concentrated under vacuum to afford (130 mg, 93%) of the
title compound (5) as a white solid, which was used without further
purification. MS m/z 353.14 [M+H].sup.+.
tert-butyl
(4-(3-((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl-
)amino)piperidine-1-carbonyl)phenyl)carbamate (6)
[0682] To a solution of
5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)-N-(piperidin-3-yl)thiazol-2-a-
mine hydrochloride (65 mg, 0.153 mmol) in DMF (0.5 mL) was added
HATU (118 mg, 0.31 mmol), 4-((tert-butoxycarbonyl)amino)benzoic
acid (72 mg, 0.31 mmol), and DIPEA (79 mg, 0.612 mmol). The
resulting mixture was stirred at room temperature for 1 hour. Then
it was diluted with EtOAc (70 mL), washed with water and brine,
dried (Na.sub.2SO.sub.4), and concentrated. The residue was
purified by an silica gel column to afford (40 mg, 43%) of the
title compound (6) as a white solid. MS m/z 572.22[M+H].sup.+.
(4-aminophenyl)(3-((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-y-
l)amino)piperidin-1-yl)methanone hydrochloride (7)
[0683] To a solution of tert-butyl
(4-(3-((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)amino)pip-
eridine-1-carbonyl)phenyl)carbamate (40 mg, 0.07 mmol) in 4 M HCl
EtOAc (1 mL). The resulting mixture was stirred at room temperature
for 30 min. Then it was concentrated under vacuum to afford (35 mg,
93%) of the title compound (7) as a white solid, which was used
without further purification. MS m/z 472.18 [M+H].sup.+.
(E)-N-(4-(3-((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2-yl)amin-
o)piperidine-1-carbonyl)phenyl)-4-dimethylamino)but-2-enamide
(B16)
[0684] To a solution of
(4-aminophenyl)(3-((5-(((5-(tert-butyl)oxazol-2-yl)methyl)thio)thiazol-2--
yl)amino)piperidin-1-yl)methanone hydrochloride (50 mg, 0.1 mmol)
in MeCN (5 mL) was added DIPEA (65 mg, 0.5 mmol) and
(E)-4-bromobut-2-enoyl chloride (22 mg, 0.12 mmol) at 0.degree. C.
for 3 min. Then it was added 4.0 M dimethylamine in THF (1 mL). The
resulting mixture was stirred at room temperature for 2 hours. Then
it was concentrated and purified by an silica gel column to afford
(10 mg, 17%) of the title compound as a white solid. MS m/z 500.18
[M+H].sup.+. .sup.1H NMR (400 MHz, MeOD) .delta. 7.72 (s, 2H), 7.41
(s, 2H), 7.01-6.88 (m, 1H), 6.74 (d, J=22.4 Hz, 2H), 3.94 (s, 2H),
3.86-3.68 (m, 4H), 3.50 (s, 1H), 3.27 (d, J=6.6 Hz, 2H), 2.44 (s,
6H), 2.23 (d, J=21.3 Hz, 1H), 2.13 (d, J=20.0 Hz, 1H), 1.98 (s,
2H), 1.72 (s, 2H), 1.45-1.40 (m, 9H).
Synthesis of B17
##STR00319##
[0685] 5-morpholinothiazol-2-amine (1)
[0686] To a mixture of 5-bromothiazol-2-amine hydrobromide (2.6 g,
10 mmol) and powdered potassium carbonate (2.77 g, 20 mmol) in NMP
(5 mL) was added morpholine (1.73 mL, 20 mmol) under argon
atmosphere and heated at 60.degree. C. for 3 hours. Reaction
mixture was cooled to room temperature and poured over ice cold
water (100 mL), extracted with EtOAc (3.times.100 mL), washed with
brine, dried over anhydrous sodium sulfate, filtered and
concentrated under reduced pressure. The residue was purified by an
silica gel column to afford (420 mg, 45.4%) of the title compound
(1) as a brown solid. MS m/z 186.07[M+H].sup.+.
3-acrylamido-N-(5-morpholinothiazol-2-yl)benzamide (B17)
[0687] To a solution of 5-morpholinothiazol-2-amine (90 mg, 0.5
mmol) in DMF (0.5 mL) was added HATU (380 mg, 1 mmol),
3-acrylamidobenzoic acid (191 mg, 1 mmol), and DIPEA (129 mg, 1
mmol). The resulting mixture was stirred at room temperature for 1
hour. Then it was diluted with EtOAc (100 mL), washed with water
and brine, dried (Na.sub.2SO.sub.4), and concentrated. The residue
was purified by an silica gel column to afford (36 mg, 21%) of the
title compound as a yellow solid. MS m/z 359.14 [M+H].sup.+.
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 12.30 (s, 1H), 10.37
(s, 1H), 8.29 (s, 1H), 7.91 (d, J=7.2 Hz, 1H), 7.78 (d, J=7.0 Hz,
1H), 7.49 (d, J=6.9 Hz, 1H), 6.78 (s, 1H), 6.47 (dd, J=15.8, 10.4
Hz, 1H), 6.31 (d, J=16.8 Hz, 1H), 5.81 (d, J=9.8 Hz, 1H), 3.75 (s,
4H), 3.05 (s, 4H).
Synthesis of B18
##STR00320##
[0688] 5-(4-methylpiperazin-1-yl)thiazol-2-amine (1)
[0689] To a mixture of 5-bromothiazol-2-amine hydrobromide (2.6 g,
10 mmol) and powdered potassium carbonate (2.77 g, 20 mmol) in NMP
(5 mL) was added 1-methylpiperazine (2.25 mL, 20 mmol) under argon
atmosphere and heated at 60.degree. C. for 3 hours. Reaction
mixture was filtered and concentrated under reduced pressure. The
residue was purified by an silica gel column to afford (340 mg,
34.3%) of the title compound (1) as a brown solid. MS m/z 199.10
[M+H].sup.+.
3-acrylamido-N-(5-(4-methylpiperazin-1-yl)thiazol-2-yl)benzamide
(B18)
[0690] To a solution of 5-(4-methylpiperazin-1-yl)thiazol-2-amine
(99 mg, 0.5 mmol) in DMF (0.5 mL) was added HATU (380 mg, 1 mmol),
3-acrylamidobenzoic acid (191 mg, 1 mmol), and DIPEA (129 mg, 1
mmol). The resulting mixture was stirred at room temperature for 1
hour. Then it was purified by an silica gel column to afford (41
mg, 22%) of the title compound as a yellow solid. MS m/z 372.17
[M+H].sup.+. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 12.22 (s,
1H), 10.35 (s, 1H), 8.28 (s, 1H), 7.91 (d, J=6.9 Hz, 1H), 7.78 (d,
J=6.7 Hz, 1H), 7.47 (t, J=7.7 Hz, 1H), 6.71 (s, 1H), 6.47 (dd,
J=16.8, 10.1 Hz, 1H), 6.31 (d, J=17.0 Hz, 1H), 5.88-5.68 (m, 1H),
3.06 (s, 4H), 2.47 (s, 4H), 2.22 (s, 3H).
Synthesis of B19
##STR00321##
[0691] 5-(4-ethylpiperazin-1-yl)thiazol-2-amine (1)
[0692] To a mixture of 5-bromothiazol-2-amine hydrobromide (2.6 g,
10 mmol) and powdered potassium carbonate (2.77 g, 20 mmol) in NMP
(5 mL) was added 1-ethylpiperazine (2.28 mL, 20 mmol) under argon
atmosphere and heated at 60.degree. C. for 3 hours. Reaction
mixture was filtered and concentrated under reduced pressure. The
residue was purified by an silica gel column to afford (382 mg,
36%) of the title compound (1) as a brown solid. MS m/z 213.11
[M+H].sup.+.
3-acrylamido-N-(5-(4-methylpiperazin-1-yl)thiazol-2-yl)benzamide
(B19)
[0693] To a solution of 5-(4-ethylpiperazin-1-yl)thiazol-2-amine
(106 mg, 0.5 mmol) in DMF (0.5 mL) was added HATU (380 mg, 1 mmol),
3-acrylamidobenzoic acid (191 mg, 1 mmol) and DIPEA (129 mg, 1
mmol). The resulting mixture was stirred at room temperature for 1
hour. Then it was purified by an silica gel column to afford (20
mg, 10.3%) of the title compound as a yellow solid. MS m/z 386.20
[M+H].sup.+. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 12.23 (s,
1H), 10.35 (s, 1H), 8.29 (s, 1H), 7.91 (d, J=7.0 Hz, 1H), 7.79 (d,
J=6.8 Hz, 1H), 7.49 (d, J=7.3 Hz, 1H), 6.72 (s, 1H), 6.58-6.43 (m,
1H), 6.31 (d, J=17.2 Hz, 1H), 5.81 (d, J=10.0 Hz, 1H), 3.07 (s,
4H), 2.52 (s, 4H), 2.38 (d, J=6.3 Hz, 2H), 1.03 (s, 3H).
Synthesis of B20
##STR00322##
[0694] tert-butyl 4-(2-aminothiazol-5-yl)piperazine-1-carboxylate
(1)
[0695] To a mixture of 5-bromothiazol-2-amine hydrobromide (2.6 g,
10 mmol) and powdered potassium carbonate (2.77 g, 20 mmol) in NMP
(5 mL) was added tert-butyl piperazine-1-carboxylate (3.72 g, 20
mmol) under argon atmosphere and heated at 60.degree. C. for 3
hours. Reaction mixture was cooled to room temperature and poured
over ice cold water (100 mL), extracted with EtOAc (3.times.100
mL), washed with brine, dried over anhydrous sodium sulfate,
filtered and concentrated under reduced pressure. The residue was
purified by an silica gel column to afford (600 mg, 42.3%) of the
title compound (1) as a brown solid. MS m/z 285.13[M+H].sup.+.
tert-butyl
4-(2-(3-acrylamidobenzamido)triazol-5-yl)piperazine-1-carboxyla- te
(B20)
[0696] To a solution of tert-butyl
4-(2-aminothiazol-5-yl)piperazine-1-carboxylate (142 mg, 0.5 mmol)
in DMF (0.5 mL) was added HATU (380 mg, 1 mmol),
3-acrylamidobenzoic acid (191 mg, 1 mmol), and DIPEA (129 mg, 1
mmol). The resulting mixture was stirred at room temperature for 1
hour. Then it was diluted with EtOAc (100 mL), washed with water
and brine, dried (Na.sub.2SO.sub.4), and concentrated. The residue
was purified by an silica gel column to afford (31 mg, 13%) of the
title compound as a yellow solid. MS m/z 458.25 [M+H].sup.+.
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 12.29 (s, 1H), 10.35
(s, 1H), 8.29 (s, 1H), 7.91 (d, J=7.5 Hz, 1H), 7.79 (d, J=7.4 Hz,
1H), 7.48 (t, J=7.9 Hz, 1H), 6.79 (s, 1H), 6.57-6.40 (m, 1H), 6.31
(d, J=16.6 Hz, 1H), 5.85-5.68 (m, 2H), 3.49 (s, 4H), 3.03 (s, 4H),
1.43 (s, 9H).
Synthesis of B22
##STR00323##
[0697] tert-butyl 1-(2-aminothiazol-5-yl)piperidin-4-ylcarbamate
(1)
[0698] To a mixture of 5-bromothiazol-2-amine hydrobromide (2.6 g,
10 mmol) and powdered potassium carbonate (2.77 g, 20 mmol) in NMP
(5 mL) was added tert-butyl piperidin-4-ylcarbamate (4.0 g, 20
mmol) under argon atmosphere and heated at 60.degree. C. for 3
hours. Reaction mixture was cooled to room temperature and poured
over ice cold water (100 mL), extracted with EtOAc (3.times.100
mL), washed with brine, dried over anhydrous sodium sulfate,
filtered and concentrated under reduced pressure. The residue was
purified by an silica gel column to afford (400 mg, 13.4%) of the
title compound (1) as a brown solid. MS m/z 299.15[M+H].sup.+.
5-(4-aminopiperidin-1-yl)thiazol-2-amine (2)
[0699] To a solution of tert-butyl
1-(2-aminothiazol-5-yl)piperidin-4-ylcarbamate (200 mg, 0.67 mmol)
in 4 M HCl EtOAc (4 mL). The resulting mixture was stirred at room
temperature for 30 min. Then it was diluted with EtOAc (100 mL).
The pH of the solution was adjusted to 7 with 1 M NaOH. The
organics washed with water and brine, dried (Na.sub.2SO.sub.4), and
concentrated to afford (120 mg, 90%) of the title compound (2) as a
slightly brown solid. MS m/z 199.10 [M+H].sup.+.
5-(4-(dimethylamino)piperidin-1-yl)thiazol-2-amine (3)
[0700] To a solution of 5-(4-aminopiperidin-1-yl)thiazol-2-amine
(120 mg, 0.61 mmol) in MeOH (5 mL) was added paraformaldehyde (90
mg, 3 mmol), AcOH (1 drop), and NaBH.sub.3CN (251 mg, 4 mmol). The
resulting mixture was stirred at room temperature for 15 hours.
Then it was concentrated under reduced pressure. The residue was
purified by an silica gel column to afford (9 mg, 5.8%) of the
title compound (3) as a brown solid. MS m/z 227.15[M+H].sup.+.
3-acrylamido-N-(5-(4-(dimethylamino)piperidin-1-yl)triazol-2-yl)benzamide
(B22)
[0701] To a solution of
5-(4-(dimethylamino)piperidin-1-yl)thiazol-2-amine (100 mg, 0.442
mmol) in DMF (0.5 mL) was added HATU (335 mg, 0.884 mmol),
3-acrylamidobenzoic acid (169 mg, 0.884 mmol), and DIPEA (228 mg,
1.768 mmol). The resulting mixture was stirred at room temperature
for 1 hour. Then it was purified by an silica gel column to afford
(35 mg, 17.5%) of the title compound as a yellow solid. MS m/z
400.20 [M+H]+. .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 12.21
(s, 1H), 10.36 (s, 1H), 8.27 (s, 1H), 7.89 (s, 1H), 7.76 (s, 1H),
7.46 (s, 1H), 6.69 (s, 1H), 6.52-6.40 (m, 1H), 6.30 (d, J=17.1 Hz,
1H), 5.79 (d, J=9.6 Hz, 1H), 3.42 (d, J=9.5 Hz, 2H), 2.74 (t,
J=11.1 Hz, 2H), 2.19 (s, 7H), 1.82 (d, J=11.2 Hz, 2H), 1.54 (d,
J=10.9 Hz, 2H).
Structure-Activity Analyses for Selected Compounds
[0702] Select compounds described herein were evaluated for
structure-activity analyses. Exemplary results are shown in Table
1.
TABLE-US-00004 TABLE 1 IC.sub.50 values of exemplary compounds
described herein. WT CDK12/ HAP1 CDK13 CDK2 CDK7 CDK9 IC.sub.50 CS
HAP1 Compound (nM) (nM) (nM) (nM) IC.sub.50 (nM) ##STR00324##
>10,000 >10,000 >10,000 >10,000 >10,000 ##STR00325##
>10,000 >10,000 >10,000 >10,000 >10,000 ##STR00326##
>10,000 >10,000 >10,000 >10,000 >10,000 ##STR00327##
>10,000 >10,000 >10,000 >10,000 >10,000 ##STR00328##
>10,000 >10,000 >10,000 >10,000 >10,000 ##STR00329##
120 6020 114 8 308 ##STR00330## 185 5550 22 ##STR00331## 174 6170
182 ##STR00332## >10,000 >10,000 >10,000 ##STR00333## 24.6
362 20A ##STR00334## 267 9230 212 ##STR00335## >10,000
>10,000 >10,000 >10,000 >10,000 ##STR00336## 15.7 557
30.9 ##STR00337## ##STR00338## 126 >10,000 164 ##STR00339## 93.5
4400 199 ##STR00340## 334 1280 66.5 119 591 ##STR00341## >1,000
>10,000 >10,000 >10,000 >10,000 ##STR00342## 22.9 3400
15.1 ##STR00343## >10,000 >10,000 >10,000 >10,000
>10,000 ##STR00344## >1,000 >10,000 >10,000 >10,000
>10,000 ##STR00345## >1,000 >10,000 >10,000 >10,000
>10,000 ##STR00346## 3170 8870 7140 >10,000 >10,000
##STR00347## 264 224 197 197 1096 ##STR00348## >1,000 >1,000
>1,000 ##STR00349## >1,000 >1,000 >1,000 ##STR00350##
>370 2870 1000 ##STR00351## >1,000 >1,000 >1,000
##STR00352## 124 447 1570 ##STR00353## 251 498 298 ##STR00354##
>1,000 >1,000 >1,000 ##STR00355## >1,000 >1,000 7280
##STR00356## 92.3 5000 65 ##STR00357## 689 158 474 ##STR00358##
9.86 242 33 ##STR00359## 11.9 230 54.5 ##STR00360## 10,000 10,000
##STR00361## 122 2111 ##STR00362## 1539 3331 ##STR00363## 7480
10,000 ##STR00364## 1180 4960 ##STR00365## 412 944 ##STR00366## 61
618 ##STR00367## ##STR00368## ##STR00369##
Genome Editing:
[0703] With regard to the exemplary results for cellular assays
shown in Table 1 above, genome editing was performed as follows.
The CRISPR/Cas9 system was used to mutate the endogenous CDK12 and
CDK13 loci to encode for CDK12 C1039S and CDK13 C1017S, both are
putative CDK12/13 inhibitor-refractory mutants. Target-specific
oligonucleotides were cloned into pX330, which carries a
codon-optimized version of Cas9 and was further modified to express
GFP for identifying transfectants. Cells were co-transfected
(X-tremeGENE 9 (Roche)) with 1) pX330 expressing Cas9 and
CDK12-targeting guide RNAs and 2) a pUC57-AMP construct bearing
1500 bp of modified CDK12 reference genome that is centered around
the CRISPR targeting site in CDK12. Two days after transfection,
cells were sorted using GFP as a marker of transfected cells and
cells were re-plated for five days. Cells were then re-plated at
low density to facilitate the isolation of individual clones.
Individual clones were isolated, expanded, and PCR genotyped using
mutant specific PCR primers. Following initial PCR screening,
individual clones were Sanger sequenced to confirm the presence of
the desired mutation. Western blot confirmed the presence of intact
CDK12 kinase. The process was sequentially repeated this time with
Cas9/sgRNA constructs to target and replace the CDK13 genetic loci.
Subsequent experiments were conducted using a CDK12 C1039S/CDK13
C1017S clone and a WT control clone that was carried through the
entirety of the CRISPR protocol but that was verified by Sanger
sequencing to be WT for CDK12 and CDK13. The genomic sequence
complementary to the CDK12-directed guide RNA that was cloned into
pX330 and used in the genome editing experiments is:
GGCAGGATTGCCATGAGTTG. The genomic sequence complementary to the
CDK13-directed guide RNA that was cloned into pX330 and used in the
genome editing experiments is: GGCAAGATTGTCATGAGTTA. The reference
genome sequence used as a repair template for CDK12 and CDK13
CRISPR was edited to 1) introduce DNA coding for serine, 2)
introduce mutations to either remove the PAM site (NGG) targeted by
CRISPR/Cas9 or introduce sufficient wobble mutations such that the
guide RNA could not recognize the repair template and thus could
not be cut by CRISPR/Cas9, and 3) introduce mutations that could
allow for mutant and WT-specific PCR amplification.
HAP1 Cell Proliferation Assay:
[0704] With regard to the exemplary results shown in Table 1 above,
the HAP1 cell proliferation assay was performed as follows. HAP1 WT
and double mutants cells were seeded at a density of 12,000
cells/well in 96-well plates. Twenty-four hours cells were then
treated with the indicated compounds in a 10-pt dose escalation
format from 1 nM to 10 .mu.M or DMSO control for 72 hrs. After 72
hrs, cells were assayed using CellTiter-Glo Luminescent Cell
Viability Assay (Promega) to determine cell viability by measuring
the amount of ATP present in each sample cell population, which is
an indicator of cell metabolic activity. Results are graphed as
fraction of the DMSO control at 72 hrs. All data points were
performed in biological triplicate.
[0705] HAP1 cells expressing putative inhibitor-refractory
mutations in CDK12 (C1039S) and CDK13 (C1017S) show up to 20-fold
less sensitivity to exemplary compounds as compared to control WT
HAP1 cells. This result indicates that a substantial portion of
intracellular compound activity comes from covalent inhibition of
CDK12 and/or CDK13 and mutation of the targeted cysteine (C1039 in
CDK12 and C1017 in CDK13) to less nucleophilic serines is
sufficient to rescue a substantial portion of anti-proliferative
activity.
EQUIVALENTS AND SCOPE
[0706] In the claims articles such as "a," "an," and "the" may mean
one or more than one unless indicated to the contrary or otherwise
evident from the context. Claims or descriptions that include "or"
between one or more members of a group are considered satisfied if
one, more than one, or all of the group members are present in,
employed in, or otherwise relevant to a given product or process
unless indicated to the contrary or otherwise evident from the
context. The invention includes embodiments in which exactly one
member of the group is present in, employed in, or otherwise
relevant to a given product or process. The invention includes
embodiments in which more than one, or all of the group members are
present in, employed in, or otherwise relevant to a given product
or process.
[0707] Furthermore, the invention encompasses all variations,
combinations, and permutations in which one or more limitations,
elements, clauses, and descriptive terms from one or more of the
listed claims is introduced into another claim. For example, any
claim that is dependent on another claim can be modified to include
one or more limitations found in any other claim that is dependent
on the same base claim. Where elements are presented as lists,
e.g., in Markush group format, each subgroup of the elements is
also disclosed, and any element(s) can be removed from the group.
It should it be understood that, in general, where the invention,
or aspects of the invention, is/are referred to as comprising
particular elements and/or features, certain embodiments of the
invention or aspects of the invention consist, or consist
essentially of, such elements and/or features. For purposes of
simplicity, those embodiments have not been specifically set forth
in haec verba herein. It is also noted that the terms "comprising"
and "containing" are intended to be open and permits the inclusion
of additional elements or steps. Where ranges are given, endpoints
are included. Furthermore, unless otherwise indicated or otherwise
evident from the context and understanding of one of ordinary skill
in the art, values that are expressed as ranges can assume any
specific value or sub-range within the stated ranges in different
embodiments of the invention, to the tenth of the unit of the lower
limit of the range, unless the context clearly dictates
otherwise.
[0708] This application refers to various issued patents, published
patent applications, journal articles, and other publications, all
of which are incorporated herein by reference. If there is a
conflict between any of the incorporated references and the instant
specification, the specification shall control. In addition, any
particular embodiment of the present invention that falls within
the prior art may be explicitly excluded from any one or more of
the claims. Because such embodiments are deemed to be known to one
of ordinary skill in the art, they may be excluded even if the
exclusion is not set forth explicitly herein. Any particular
embodiment of the invention can be excluded from any claim, for any
reason, whether or not related to the existence of prior art.
[0709] Those skilled in the art will recognize or be able to
ascertain using no more than routine experimentation many
equivalents to the specific embodiments described herein. The scope
of the present embodiments described herein is not intended to be
limited to the above Description, but rather is as set forth in the
appended claims. Those of ordinary skill in the art will appreciate
that various changes and modifications to this description may be
made without departing from the spirit or scope of the present
invention, as defined in the following claims.
Sequence CWU 1
1
51346PRTHomo sapiens 1Met Ala Leu Asp Val Lys Ser Arg Ala Lys Arg
Tyr Glu Lys Leu Asp 1 5 10 15 Phe Leu Gly Glu Gly Gln Phe Ala Thr
Val Tyr Lys Ala Arg Asp Lys 20 25 30 Asn Thr Asn Gln Ile Val Ala
Ile Lys Lys Ile Lys Leu Gly His Arg 35 40 45 Ser Glu Ala Lys Asp
Gly Ile Asn Arg Thr Ala Leu Arg Glu Ile Lys 50 55 60 Leu Leu Gln
Glu Leu Ser His Pro Asn Ile Ile Gly Leu Leu Asp Ala 65 70 75 80 Phe
Gly His Lys Ser Asn Ile Ser Leu Val Phe Asp Phe Met Glu Thr 85 90
95 Asp Leu Glu Val Ile Ile Lys Asp Asn Ser Leu Val Leu Thr Pro Ser
100 105 110 His Ile Lys Ala Tyr Met Leu Met Thr Leu Gln Gly Leu Glu
Tyr Leu 115 120 125 His Gln His Trp Ile Leu His Arg Asp Leu Lys Pro
Asn Asn Leu Leu 130 135 140 Leu Asp Glu Asn Gly Val Leu Lys Leu Ala
Asp Phe Gly Leu Ala Lys 145 150 155 160 Ser Phe Gly Ser Pro Asn Arg
Ala Tyr Thr His Gln Val Val Thr Arg 165 170 175 Trp Tyr Arg Ala Pro
Glu Leu Leu Phe Gly Ala Arg Met Tyr Gly Val 180 185 190 Gly Val Asp
Met Trp Ala Val Gly Cys Ile Leu Ala Glu Leu Leu Leu 195 200 205 Arg
Val Pro Phe Leu Pro Gly Asp Ser Asp Leu Asp Gln Leu Thr Arg 210 215
220 Ile Phe Glu Thr Leu Gly Thr Pro Thr Glu Glu Gln Trp Pro Asp Met
225 230 235 240 Cys Ser Leu Pro Asp Tyr Val Thr Phe Lys Ser Phe Pro
Gly Ile Pro 245 250 255 Leu His His Ile Phe Ser Ala Ala Gly Asp Asp
Leu Leu Asp Leu Ile 260 265 270 Gln Gly Leu Phe Leu Phe Asn Pro Cys
Ala Arg Ile Thr Ala Thr Gln 275 280 285 Ala Leu Lys Met Lys Tyr Phe
Ser Asn Arg Pro Gly Pro Thr Pro Gly 290 295 300 Cys Gln Leu Pro Arg
Pro Asn Cys Pro Val Glu Thr Leu Lys Glu Gln 305 310 315 320 Ser Asn
Pro Ala Leu Ala Ile Lys Arg Lys Arg Thr Glu Ala Leu Glu 325 330 335
Gln Gly Gly Leu Pro Lys Lys Leu Ile Phe 340 345 21490PRTHomo
sapiens 2Met Pro Asn Ser Glu Arg His Gly Gly Lys Lys Asp Gly Ser
Gly Gly 1 5 10 15 Ala Ser Gly Thr Leu Gln Pro Ser Ser Gly Gly Gly
Ser Ser Asn Ser 20 25 30 Arg Glu Arg His Arg Leu Val Ser Lys His
Lys Arg His Lys Ser Lys 35 40 45 His Ser Lys Asp Met Gly Leu Val
Thr Pro Glu Ala Ala Ser Leu Gly 50 55 60 Thr Val Ile Lys Pro Leu
Val Glu Tyr Asp Asp Ile Ser Ser Asp Ser 65 70 75 80 Asp Thr Phe Ser
Asp Asp Met Ala Phe Lys Leu Asp Arg Arg Glu Asn 85 90 95 Asp Glu
Arg Arg Gly Ser Asp Arg Ser Asp Arg Leu His Lys His Arg 100 105 110
His His Gln His Arg Arg Ser Arg Asp Leu Leu Lys Ala Lys Gln Thr 115
120 125 Glu Lys Glu Lys Ser Gln Glu Val Ser Ser Lys Ser Gly Ser Met
Lys 130 135 140 Asp Arg Ile Ser Gly Ser Ser Lys Arg Ser Asn Glu Glu
Thr Asp Asp 145 150 155 160 Tyr Gly Lys Ala Gln Val Ala Lys Ser Ser
Ser Lys Glu Ser Arg Ser 165 170 175 Ser Lys Leu His Lys Glu Lys Thr
Arg Lys Glu Arg Glu Leu Lys Ser 180 185 190 Gly His Lys Asp Arg Ser
Lys Ser His Arg Lys Arg Glu Thr Pro Lys 195 200 205 Ser Tyr Lys Thr
Val Asp Ser Pro Lys Arg Arg Ser Arg Ser Pro His 210 215 220 Arg Lys
Trp Ser Asp Ser Ser Lys Gln Asp Asp Ser Pro Ser Gly Ala 225 230 235
240 Ser Tyr Gly Gln Asp Tyr Asp Leu Ser Pro Ser Arg Ser His Thr Ser
245 250 255 Ser Asn Tyr Asp Ser Tyr Lys Lys Ser Pro Gly Ser Thr Ser
Arg Arg 260 265 270 Gln Ser Val Ser Pro Pro Tyr Lys Glu Pro Ser Ala
Tyr Gln Ser Ser 275 280 285 Thr Arg Ser Pro Ser Pro Tyr Ser Arg Arg
Gln Arg Ser Val Ser Pro 290 295 300 Tyr Ser Arg Arg Arg Ser Ser Ser
Tyr Glu Arg Ser Gly Ser Tyr Ser 305 310 315 320 Gly Arg Ser Pro Ser
Pro Tyr Gly Arg Arg Arg Ser Ser Ser Pro Phe 325 330 335 Leu Ser Lys
Arg Ser Leu Ser Arg Ser Pro Leu Pro Ser Arg Lys Ser 340 345 350 Met
Lys Ser Arg Ser Arg Ser Pro Ala Tyr Ser Arg His Ser Ser Ser 355 360
365 His Ser Lys Lys Lys Arg Ser Ser Ser Arg Ser Arg His Ser Ser Ile
370 375 380 Ser Pro Val Arg Leu Pro Leu Asn Ser Ser Leu Gly Ala Glu
Leu Ser 385 390 395 400 Arg Lys Lys Lys Glu Arg Ala Ala Ala Ala Ala
Ala Ala Lys Met Asp 405 410 415 Gly Lys Glu Ser Lys Gly Ser Pro Val
Phe Leu Pro Arg Lys Glu Asn 420 425 430 Ser Ser Val Glu Ala Lys Asp
Ser Gly Leu Glu Ser Lys Lys Leu Pro 435 440 445 Arg Ser Val Lys Leu
Glu Lys Ser Ala Pro Asp Thr Glu Leu Val Asn 450 455 460 Val Thr His
Leu Asn Thr Glu Val Lys Asn Ser Ser Asp Thr Gly Lys 465 470 475 480
Val Lys Leu Asp Glu Asn Ser Glu Lys His Leu Val Lys Asp Leu Lys 485
490 495 Ala Gln Gly Thr Arg Asp Ser Lys Pro Ile Ala Leu Lys Glu Glu
Ile 500 505 510 Val Thr Pro Lys Glu Thr Glu Thr Ser Glu Lys Glu Thr
Pro Pro Pro 515 520 525 Leu Pro Thr Ile Ala Ser Pro Pro Pro Pro Leu
Pro Thr Thr Thr Pro 530 535 540 Pro Pro Gln Thr Pro Pro Leu Pro Pro
Leu Pro Pro Ile Pro Ala Leu 545 550 555 560 Pro Gln Gln Pro Pro Leu
Pro Pro Ser Gln Pro Ala Phe Ser Gln Val 565 570 575 Pro Ala Ser Ser
Thr Ser Thr Leu Pro Pro Ser Thr His Ser Lys Thr 580 585 590 Ser Ala
Val Ser Ser Gln Ala Asn Ser Gln Pro Pro Val Gln Val Ser 595 600 605
Val Lys Thr Gln Val Ser Val Thr Ala Ala Ile Pro His Leu Lys Thr 610
615 620 Ser Thr Leu Pro Pro Leu Pro Leu Pro Pro Leu Leu Pro Gly Asp
Asp 625 630 635 640 Asp Met Asp Ser Pro Lys Glu Thr Leu Pro Ser Lys
Pro Val Lys Lys 645 650 655 Glu Lys Glu Gln Arg Thr Arg His Leu Leu
Thr Asp Leu Pro Leu Pro 660 665 670 Pro Glu Leu Pro Gly Gly Asp Leu
Ser Pro Pro Asp Ser Pro Glu Pro 675 680 685 Lys Ala Ile Thr Pro Pro
Gln Gln Pro Tyr Lys Lys Arg Pro Lys Ile 690 695 700 Cys Cys Pro Arg
Tyr Gly Glu Arg Arg Gln Thr Glu Ser Asp Trp Gly 705 710 715 720 Lys
Arg Cys Val Asp Lys Phe Asp Ile Ile Gly Ile Ile Gly Glu Gly 725 730
735 Thr Tyr Gly Gln Val Tyr Lys Ala Lys Asp Lys Asp Thr Gly Glu Leu
740 745 750 Val Ala Leu Lys Lys Val Arg Leu Asp Asn Glu Lys Glu Gly
Phe Pro 755 760 765 Ile Thr Ala Ile Arg Glu Ile Lys Ile Leu Arg Gln
Leu Ile His Arg 770 775 780 Ser Val Val Asn Met Lys Glu Ile Val Thr
Asp Lys Gln Asp Ala Leu 785 790 795 800 Asp Phe Lys Lys Asp Lys Gly
Ala Phe Tyr Leu Val Phe Glu Tyr Met 805 810 815 Asp His Asp Leu Met
Gly Leu Leu Glu Ser Gly Leu Val His Phe Ser 820 825 830 Glu Asp His
Ile Lys Ser Phe Met Lys Gln Leu Met Glu Gly Leu Glu 835 840 845 Tyr
Cys His Lys Lys Asn Phe Leu His Arg Asp Ile Lys Cys Ser Asn 850 855
860 Ile Leu Leu Asn Asn Ser Gly Gln Ile Lys Leu Ala Asp Phe Gly Leu
865 870 875 880 Ala Arg Leu Tyr Asn Ser Glu Glu Ser Arg Pro Tyr Thr
Asn Lys Val 885 890 895 Ile Thr Leu Trp Tyr Arg Pro Pro Glu Leu Leu
Leu Gly Glu Glu Arg 900 905 910 Tyr Thr Pro Ala Ile Asp Val Trp Ser
Cys Gly Cys Ile Leu Gly Glu 915 920 925 Leu Phe Thr Lys Lys Pro Ile
Phe Gln Ala Asn Leu Glu Leu Ala Gln 930 935 940 Leu Glu Leu Ile Ser
Arg Leu Cys Gly Ser Pro Cys Pro Ala Val Trp 945 950 955 960 Pro Asp
Val Ile Lys Leu Pro Tyr Phe Asn Thr Met Lys Pro Lys Lys 965 970 975
Gln Tyr Arg Arg Arg Leu Arg Glu Glu Phe Ser Phe Ile Pro Ser Ala 980
985 990 Ala Leu Asp Leu Leu Asp His Met Leu Thr Leu Asp Pro Ser Lys
Arg 995 1000 1005 Cys Thr Ala Glu Gln Thr Leu Gln Ser Asp Phe Leu
Lys Asp Val 1010 1015 1020 Glu Leu Ser Lys Met Ala Pro Pro Asp Leu
Pro His Trp Gln Asp 1025 1030 1035 Cys His Glu Leu Trp Ser Lys Lys
Arg Arg Arg Gln Arg Gln Ser 1040 1045 1050 Gly Val Val Val Glu Glu
Pro Pro Pro Ser Lys Thr Ser Arg Lys 1055 1060 1065 Glu Thr Thr Ser
Gly Thr Ser Thr Glu Pro Val Lys Asn Ser Ser 1070 1075 1080 Pro Ala
Pro Pro Gln Pro Ala Pro Gly Lys Val Glu Ser Gly Ala 1085 1090 1095
Gly Asp Ala Ile Gly Leu Ala Asp Ile Thr Gln Gln Leu Asn Gln 1100
1105 1110 Ser Glu Leu Ala Val Leu Leu Asn Leu Leu Gln Ser Gln Thr
Asp 1115 1120 1125 Leu Ser Ile Pro Gln Met Ala Gln Leu Leu Asn Ile
His Ser Asn 1130 1135 1140 Pro Glu Met Gln Gln Gln Leu Glu Ala Leu
Asn Gln Ser Ile Ser 1145 1150 1155 Ala Leu Thr Glu Ala Thr Ser Gln
Gln Gln Asp Ser Glu Thr Met 1160 1165 1170 Ala Pro Glu Glu Ser Leu
Lys Glu Ala Pro Ser Ala Pro Val Ile 1175 1180 1185 Leu Pro Ser Ala
Glu Gln Thr Thr Leu Glu Ala Ser Ser Thr Pro 1190 1195 1200 Ala Asp
Met Gln Asn Ile Leu Ala Val Leu Leu Ser Gln Leu Met 1205 1210 1215
Lys Thr Gln Glu Pro Ala Gly Ser Leu Glu Glu Asn Asn Ser Asp 1220
1225 1230 Lys Asn Ser Gly Pro Gln Gly Pro Arg Arg Thr Pro Thr Met
Pro 1235 1240 1245 Gln Glu Glu Ala Ala Ala Cys Pro Pro His Ile Leu
Pro Pro Glu 1250 1255 1260 Lys Arg Pro Pro Glu Pro Pro Gly Pro Pro
Pro Pro Pro Pro Pro 1265 1270 1275 Pro Pro Leu Val Glu Gly Asp Leu
Ser Ser Ala Pro Gln Glu Leu 1280 1285 1290 Asn Pro Ala Val Thr Ala
Ala Leu Leu Gln Leu Leu Ser Gln Pro 1295 1300 1305 Glu Ala Glu Pro
Pro Gly His Leu Pro His Glu His Gln Ala Leu 1310 1315 1320 Arg Pro
Met Glu Tyr Ser Thr Arg Pro Arg Pro Asn Arg Thr Tyr 1325 1330 1335
Gly Asn Thr Asp Gly Pro Glu Thr Gly Phe Ser Ala Ile Asp Thr 1340
1345 1350 Asp Glu Arg Asn Ser Gly Pro Ala Leu Thr Glu Ser Leu Val
Gln 1355 1360 1365 Thr Leu Val Lys Asn Arg Thr Phe Ser Gly Ser Leu
Ser His Leu 1370 1375 1380 Gly Glu Ser Ser Ser Tyr Gln Gly Thr Gly
Ser Val Gln Phe Pro 1385 1390 1395 Gly Asp Gln Asp Leu Arg Phe Ala
Arg Val Pro Leu Ala Leu His 1400 1405 1410 Pro Val Val Gly Gln Pro
Phe Leu Lys Ala Glu Gly Ser Ser Asn 1415 1420 1425 Ser Val Val His
Ala Glu Thr Lys Leu Gln Asn Tyr Gly Glu Leu 1430 1435 1440 Gly Pro
Gly Thr Thr Gly Ala Ser Ser Ser Gly Ala Gly Leu His 1445 1450 1455
Trp Gly Gly Pro Thr Gln Ser Ser Ala Tyr Gly Lys Leu Tyr Arg 1460
1465 1470 Gly Pro Thr Arg Val Pro Pro Arg Gly Gly Arg Gly Arg Gly
Val 1475 1480 1485 Pro Tyr 1490 31512PRTHomo sapiens 3Met Pro Ser
Ser Ser Asp Thr Ala Leu Gly Gly Gly Gly Gly Leu Ser 1 5 10 15 Trp
Ala Glu Lys Lys Leu Glu Glu Arg Arg Lys Arg Arg Arg Phe Leu 20 25
30 Ser Pro Gln Gln Pro Pro Leu Leu Leu Pro Leu Leu Gln Pro Gln Leu
35 40 45 Leu Gln Pro Pro Pro Pro Pro Pro Pro Leu Leu Phe Leu Ala
Ala Pro 50 55 60 Gly Thr Ala Ala Ala Ala Ala Ala Ala Ala Ala Ala
Ser Ser Ser Cys 65 70 75 80 Phe Ser Pro Gly Pro Pro Leu Glu Val Lys
Arg Leu Ala Arg Gly Lys 85 90 95 Arg Arg Ala Gly Gly Arg Gln Lys
Arg Arg Arg Gly Pro Arg Ala Gly 100 105 110 Gln Glu Ala Glu Lys Arg
Arg Val Phe Ser Leu Pro Gln Pro Gln Gln 115 120 125 Asp Gly Gly Gly
Gly Ala Ser Ser Gly Gly Gly Val Thr Pro Leu Val 130 135 140 Glu Tyr
Glu Asp Val Ser Ser Gln Ser Glu Gln Gly Leu Leu Leu Gly 145 150 155
160 Gly Ala Ser Ala Ala Thr Ala Ala Thr Ala Ala Gly Gly Thr Gly Gly
165 170 175 Ser Gly Gly Ser Pro Ala Ser Ser Ser Gly Thr Gln Arg Arg
Gly Glu 180 185 190 Gly Ser Glu Arg Arg Pro Arg Arg Asp Arg Arg Ser
Ser Ser Gly Arg 195 200 205 Ser Lys Glu Arg His Arg Glu His Arg Arg
Arg Asp Gly Gln Arg Gly 210 215 220 Gly Ser Glu Ala Ser Lys Ser Arg
Ser Arg His Ser His Ser Gly Glu 225 230 235 240 Glu Arg Ala Glu Val
Ala Lys Ser Gly Ser Ser Ser Ser Ser Gly Gly 245 250 255 Arg Arg Lys
Ser Ala Ser Ala Thr Ser Ser Ser Ser Ser Ser Arg Lys 260 265 270 Asp
Arg Asp Ser Lys Ala His Arg Ser Arg Thr Lys Ser Ser Lys Glu 275 280
285 Pro Pro Ser Ala Tyr Lys Glu Pro Pro Lys Ala Tyr Arg Glu Asp Lys
290 295 300 Thr Glu Pro Lys Ala Tyr Arg Arg Arg Arg Ser Leu Ser Pro
Leu Gly 305 310 315 320 Gly Arg Asp Asp Ser Pro Val Ser His Arg Ala
Ser Gln Ser Leu Arg 325 330 335 Ser Arg Lys Ser Pro Ser Pro Ala Gly
Gly Gly Ser Ser Pro Tyr Ser 340 345 350 Arg Arg Leu Pro Arg Ser Pro
Ser Pro Tyr Ser Arg Arg Arg Ser Pro 355 360 365 Ser Tyr Ser Arg His
Ser Ser Tyr Glu Arg Gly Gly Asp Val Ser Pro 370 375 380 Ser Pro Tyr
Ser Ser Ser Ser Trp Arg Arg Ser Arg Ser Pro Tyr Ser 385 390 395 400
Pro Val Leu Arg Arg Ser Gly Lys Ser Arg Ser Arg Ser Pro Tyr Ser 405
410 415 Ser Arg His Ser Arg Ser Arg Ser Arg His Arg Leu Ser Arg Ser
Arg 420 425 430 Ser Arg His Ser Ser Ile Ser Pro Ser Thr Leu Thr
Leu
Lys Ser Ser 435 440 445 Leu Ala Ala Glu Leu Asn Lys Asn Lys Lys Ala
Arg Ala Ala Glu Ala 450 455 460 Ala Arg Ala Ala Glu Ala Ala Lys Ala
Ala Glu Ala Thr Lys Ala Ala 465 470 475 480 Glu Ala Ala Ala Lys Ala
Ala Lys Ala Ser Asn Thr Ser Thr Pro Thr 485 490 495 Lys Gly Asn Thr
Glu Thr Ser Ala Ser Ala Ser Gln Thr Asn His Val 500 505 510 Lys Asp
Val Lys Lys Ile Lys Ile Glu His Ala Pro Ser Pro Ser Ser 515 520 525
Gly Gly Thr Leu Lys Asn Asp Lys Ala Lys Thr Lys Pro Pro Leu Gln 530
535 540 Val Thr Lys Val Glu Asn Asn Leu Ile Val Asp Lys Ala Thr Lys
Lys 545 550 555 560 Ala Val Ile Val Gly Lys Glu Ser Lys Ser Ala Ala
Thr Lys Glu Glu 565 570 575 Ser Val Ser Leu Lys Glu Lys Thr Lys Pro
Leu Thr Pro Ser Ile Gly 580 585 590 Ala Lys Glu Lys Glu Gln His Val
Ala Leu Val Thr Ser Thr Leu Pro 595 600 605 Pro Leu Pro Leu Pro Pro
Met Leu Pro Glu Asp Lys Glu Ala Asp Ser 610 615 620 Leu Arg Gly Asn
Ile Ser Val Lys Ala Val Lys Lys Glu Val Glu Lys 625 630 635 640 Lys
Leu Arg Cys Leu Leu Ala Asp Leu Pro Leu Pro Pro Glu Leu Pro 645 650
655 Gly Gly Asp Asp Leu Ser Lys Ser Pro Glu Glu Lys Lys Thr Ala Thr
660 665 670 Gln Leu His Ser Lys Arg Arg Pro Lys Ile Cys Gly Pro Arg
Tyr Gly 675 680 685 Glu Thr Lys Glu Lys Asp Ile Asp Trp Gly Lys Arg
Cys Val Asp Lys 690 695 700 Phe Asp Ile Ile Gly Ile Ile Gly Glu Gly
Thr Tyr Gly Gln Val Tyr 705 710 715 720 Lys Ala Arg Asp Lys Asp Thr
Gly Glu Met Val Ala Leu Lys Lys Val 725 730 735 Arg Leu Asp Asn Glu
Lys Glu Gly Phe Pro Ile Thr Ala Ile Arg Glu 740 745 750 Ile Lys Ile
Leu Arg Gln Leu Thr His Gln Ser Ile Ile Asn Met Lys 755 760 765 Glu
Ile Val Thr Asp Lys Glu Asp Ala Leu Asp Phe Lys Lys Asp Lys 770 775
780 Gly Ala Phe Tyr Leu Val Phe Glu Tyr Met Asp His Asp Leu Met Gly
785 790 795 800 Leu Leu Glu Ser Gly Leu Val His Phe Asn Glu Asn His
Ile Lys Ser 805 810 815 Phe Met Arg Gln Leu Met Glu Gly Leu Asp Tyr
Cys His Lys Lys Asn 820 825 830 Phe Leu His Arg Asp Ile Lys Cys Ser
Asn Ile Leu Leu Asn Asn Arg 835 840 845 Gly Gln Ile Lys Leu Ala Asp
Phe Gly Leu Ala Arg Leu Tyr Ser Ser 850 855 860 Glu Glu Ser Arg Pro
Tyr Thr Asn Lys Val Ile Thr Leu Trp Tyr Arg 865 870 875 880 Pro Pro
Glu Leu Leu Leu Gly Glu Glu Arg Tyr Thr Pro Ala Ile Asp 885 890 895
Val Trp Ser Cys Gly Cys Ile Leu Gly Glu Leu Phe Thr Lys Lys Pro 900
905 910 Ile Phe Gln Ala Asn Gln Glu Leu Ala Gln Leu Glu Leu Ile Ser
Arg 915 920 925 Ile Cys Gly Ser Pro Cys Pro Ala Val Trp Pro Asp Val
Ile Lys Leu 930 935 940 Pro Tyr Phe Asn Thr Met Lys Pro Lys Lys Gln
Tyr Arg Arg Lys Leu 945 950 955 960 Arg Glu Glu Phe Val Phe Ile Pro
Ala Ala Ala Leu Asp Leu Phe Asp 965 970 975 Tyr Met Leu Ala Leu Asp
Pro Ser Lys Arg Cys Thr Ala Glu Gln Ala 980 985 990 Leu Gln Cys Glu
Phe Leu Arg Asp Val Glu Pro Ser Lys Met Pro Pro 995 1000 1005 Pro
Asp Leu Pro Leu Trp Gln Asp Cys His Glu Leu Trp Ser Lys 1010 1015
1020 Lys Arg Arg Arg Gln Lys Gln Met Gly Met Thr Asp Asp Val Ser
1025 1030 1035 Thr Ile Lys Ala Pro Arg Lys Asp Leu Ser Leu Gly Leu
Asp Asp 1040 1045 1050 Ser Arg Thr Asn Thr Pro Gln Gly Val Leu Pro
Ser Ser Gln Leu 1055 1060 1065 Lys Ser Gln Gly Ser Ser Asn Val Ala
Pro Val Lys Thr Gly Pro 1070 1075 1080 Gly Gln His Leu Asn His Ser
Glu Leu Ala Ile Leu Leu Asn Leu 1085 1090 1095 Leu Gln Ser Lys Thr
Ser Val Asn Met Ala Asp Phe Val Gln Val 1100 1105 1110 Leu Asn Ile
Lys Val Asn Ser Glu Thr Gln Gln Gln Leu Asn Lys 1115 1120 1125 Ile
Asn Leu Pro Ala Gly Ile Leu Ala Thr Gly Glu Lys Gln Thr 1130 1135
1140 Asp Pro Ser Thr Pro Gln Gln Glu Ser Ser Lys Pro Leu Gly Gly
1145 1150 1155 Ile Gln Pro Ser Ser Gln Thr Ile Gln Pro Lys Val Glu
Thr Asp 1160 1165 1170 Ala Ala Gln Ala Ala Val Gln Ser Ala Phe Ala
Val Leu Leu Thr 1175 1180 1185 Gln Leu Ile Lys Ala Gln Gln Ser Lys
Gln Lys Asp Val Leu Leu 1190 1195 1200 Glu Glu Arg Glu Asn Gly Ser
Gly His Glu Ala Ser Leu Gln Leu 1205 1210 1215 Arg Pro Pro Pro Glu
Pro Ser Thr Pro Val Ser Gly Gln Asp Asp 1220 1225 1230 Leu Ile Gln
His Gln Asp Met Arg Ile Leu Glu Leu Thr Pro Glu 1235 1240 1245 Pro
Asp Arg Pro Arg Ile Leu Pro Pro Asp Gln Arg Pro Pro Glu 1250 1255
1260 Pro Pro Glu Pro Pro Pro Val Thr Glu Glu Asp Leu Asp Tyr Arg
1265 1270 1275 Thr Glu Asn Gln His Val Pro Thr Thr Ser Ser Ser Leu
Thr Asp 1280 1285 1290 Pro His Ala Gly Val Lys Ala Ala Leu Leu Gln
Leu Leu Ala Gln 1295 1300 1305 His Gln Pro Gln Asp Asp Pro Lys Arg
Glu Gly Gly Ile Asp Tyr 1310 1315 1320 Gln Ala Gly Asp Thr Tyr Val
Ser Thr Ser Asp Tyr Lys Asp Asn 1325 1330 1335 Phe Gly Ser Ser Ser
Phe Ser Ser Ala Pro Tyr Val Ser Asn Asp 1340 1345 1350 Gly Leu Gly
Ser Ser Ser Ala Pro Pro Leu Glu Arg Arg Ser Phe 1355 1360 1365 Ile
Gly Asn Ser Asp Ile Gln Ser Leu Asp Asn Tyr Ser Thr Ala 1370 1375
1380 Ser Ser His Ser Gly Gly Pro Pro Gln Pro Ser Ala Phe Ser Glu
1385 1390 1395 Ser Phe Pro Ser Ser Val Ala Gly Tyr Gly Asp Ile Tyr
Leu Asn 1400 1405 1410 Ala Gly Pro Met Leu Phe Ser Gly Asp Lys Asp
His Arg Phe Glu 1415 1420 1425 Tyr Ser His Gly Pro Ile Ala Val Leu
Ala Asn Ser Ser Asp Pro 1430 1435 1440 Ser Thr Gly Pro Glu Ser Thr
His Pro Leu Pro Ala Lys Met His 1445 1450 1455 Asn Tyr Asn Tyr Gly
Gly Asn Leu Gln Glu Asn Pro Ser Gly Pro 1460 1465 1470 Ser Leu Met
His Gly Gln Thr Trp Thr Ser Pro Ala Gln Gly Pro 1475 1480 1485 Gly
Tyr Ser Gln Gly Tyr Arg Gly His Ile Ser Thr Ser Thr Gly 1490 1495
1500 Arg Gly Arg Gly Arg Gly Leu Pro Tyr 1505 1510 420DNAArtificial
SequenceSynthetic Polynucleotide 4ggcaggattg ccatgagttg
20520DNAArtificial SequenceSynthetic Polynucleotide 5ggcaagattg
tcatgagtta 20
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